IV ... MYCOTAXON 132(2)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-TWO (2) — TABLE OF CONTENTS

132-2: TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS & EDITORIALS

REVIG WEIS eS Se eile By Packs Ane, WILE, SSD A tial Neh, SARENN SOD, SNe AN vi

Nomenclatural novelties: ty pifiCatt0ns. nce. cas seo oe le ee ya ete ew vii

PELE PB Sad BIN ee sta Se nett! RR! 4 teahy nen Ser OTe Bh tae, Viii

ETOP HE TEAIEOE 35 Hak i 8 eo ao ge Woe ak 2 at asd OA ule cool otto) Ee nea He, ix

In remembrance: Gams, Mel'nik, de Souza, Vasilyeva...... 0.0.0 cece ee eee xi

RESEARCH ARTICLES

Three species of Linkosia and Spadicoides new to China

ZHAO-HUAN XU, DIAN-MING Hu, You-QIANG LUO & JIAN Ma 243

Ascomycetous fungi of Siberia. 3. Elytroderma baikalense sp. nov.

Larissa N. VASILYEVAT, TATYANA I. MOROZOVA & STEVEN L. STEPHENSON 251

Inocybe ahmadii sp. nov. and a new record of I. leptocystis

from Pakistan AYESHA Farooq], Fauzia AQDUS,

ABDUL REHMAN NIAZI, SANA JABEEN & ABDUL NASIR KHALID 257

Corynespora titarpaniensis sp. nov. on Lepidagathis sp.

from Central India PRAKASH KUSHWAHA,

RAGHVENDRA SINGH & SHAMBHU KUMAR 271

Morganella austromontana sp. nov. from the

South Brazilian Plateau CAMILA R. ALVES,

VAGNER G. CORTEZ & Rosa M.B. DA SILVEIRA 281

Microidium phyllanthi-reticulati, sp. nov.

on Phyllanthus reticulatus JAMJAN MEEBOON & SUSUMU TAKAMATSU 289

Xylaria nelumboniformis sp. nov.

from tropical China Har-x1a Ma, Larissa VASILYEVAT & Yu Li 299

Neozygites linanensis sp. nov., a fungal pathogen

infecting bamboo aphids in southeast China XIANG ZHOU,

CrisTIAN MONTALVA, NOLBERTO ARISMENDI & FEI Hone 305

Five Lecidea lichens new to China XIANG- XIANG ZHAO,

ZUN-TIAN ZHAO, CONG-CONG MIAO, ZHAO-JIE REN & Lu-LU ZHANG 317

Pseudobaeospora lilacina sp. nov., the first report

of the genus from China SHUANG-YAN WJ, J1A-JrA L1,

MING ZHANG, X1A0-DaN Yu, SHuU-X1A Lv & DoNG-Xu Cao 327

APRIL-JUNE 2017... V

Gymnopilus dilepis, a new record in Thailand NAKARIN SUWANNARACH,

JATURONG KUMLA, KRIANGSAK SRI-NGERNYUANG & SAISAMORN LUMYONG 337

A reassessment of Hourangia cheoi from Yunnan, China

MarTTEO GELARDI, ALFREDO VIZZINI & ENRICO ERCOLE 343

Bharatheeya coronata sp. nov., a conidial fungus from Brazil

Lucas BARBOSA CONCEICAO, MARCOS FABIO OLIVEIRA MARQUES,

JOSIANE SANTANA MONTEIRO, Luis FERNANDO PASCHOLATI GUSMAO,

FLAVIA RODRIGUES BARBOSA & RAFAEL F, CASTANEDA-RUIZ 357

New records of Phallales from Paraguay MICHELLE GERALDINE CAMPI GAONA,

LARISSA TRIERVEILER-PEREIRA & YANINE ELIZABETH MAUBET CANO 361

Arthrotaeniolella aquatica gen. & sp. nov. and

Pseudospiropes piatanensis sp. nov. from Brazil

JOSIANE SANTANA MONTEIRO, LUCAS BARBOSA CONCEICAO,

Luis FERNANDO PASCHOLATI GUSMAO & RAFAEL FE, CASTANEDA-Ru1z 373

Cunninghamella clavata from Brazil: a new record

for the western hemisphere Ana L.S DE M. ALVES,

CaRLos A.F. DE SOUZA, RAFAEL J.V. DE OLIVEIRA,

THALLINE R.L. CORDEIRO & ANDRE L.C.M. DE A. SANTIAGO 381

A comparative SEM-study of morphological characters

in Cribraria José Martin Ram{irez-OrTEGA,

ARTURO ESTRADA-TORRES & EFRAIN DE Luna: 391

New records of Ascomycota from tropical dry forest

in Sonora, Mexico TANIA RAYMUNDO, MARTHA L. CORONADO,

ALDO GUTIERREZ, MARTIN ESQUEDA & RICARDO VALENZUELA 421

Dictyotrichocladium aquaticum gen. & sp. nov. and

Minimelanolocus aquatilis sp. nov. from freshwater

in Brazil’s semiarid region PaTRIiCIA OLIVEIRA FIUZA,

Lucas BARBOSA CONCEICAO, MARCOS FABIO OLIVEIRA MARQUES,

Luis FERNANDO PASCHOLATI GUSMAO & RAFAEL F, CASTANEDA-Ru1z 433

Lecanora shangrilaensis sp. nov., on pinecones from China

Ler LU & ZuN-TIAN ZHAO 441

Cephaliophora tropica: third European record

MALGORZATA RUSZKIEWICZ-MICHALSKA, PIOTR KNYSAK, IZABELA SKROBEK,

ANETA GWIAZDA, SEBASTIAN PISKORSKI & JOANNA ZELAZNA-WIECZOREK 445

Tretoheliocephala compacta gen. & sp. nov. from

the Brazilian semi-arid region Luis FERNANDO PAscHOLATI GUSMAO,

JOSIANE SANTANA MONTEIRO & RAFAEL F. CASTANEDA-Rut1z 453

Sporormiella longicolla sp. nov. and

new Sporormiella records on herbivore dung from Brazil

ROGER FAGNER RIBEIRO MELO, ANDREW N. MILLER & LEONOR Costa Mata 459

VI ... MYCOTAXON 132(2)

REVIEWERS — VOLUME ONE HUNDRED THIRTY-TWO (2)

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 issue.

Marek Barta

Turi Goulart Baseia

Jose Luiz Bezerra

Uwe Braun

Marcela Caceres

Cecilia C. Carmaran

Rafael F. Castafteda-Ruiz

Balint Dima

Maria Martha Dios

Maria Dynowska

Ming-Guang Feng

Patricia Oliveira Fiuza

Harish C. Gugnani

Shouyu Guo

Maria Luciana Hernandez-Caffot

Alfredo Justo

Sergey Kondratyuk

Irmgard Krisai-Greilhuber

De-Wei Li

Tai-hui Li

Jian Ma

Li-Guo Ma

José G. Marmolejo

Eric H.C. McKenzie

Lorelei L. Norvell

Yuri Novozhilov

Shaun R. Pennycook

Jadergudson Pereira

Donald H. Pfister

Orlando Popoff

Gerhard Rambold

Martin Ryberg

H.D. Shin

Giampaolo Simonini

José Ivanildo de Souzat

Steven L. Stephenson

A.J.S. Whalley

Hai-Sheng Yuan

Nian-Kai Zeng

Xiu-Guo Zhang

Wen- Ying Zhuang

APRIL-JUNE 2017... VII

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 132(2)

Arthrotaeniolella L.B. Conc., M.E.O. Marques, J.S. Monteiro,

Gusmao & R.F. Castafieda [MB 821602], p. 374

Arthrotaeniolella aquatica L.B. Conc., M.F.O. Marques, J.S. Monteiro,

Gusmao & R.F. Castafieda [MB 821603], p. 374

Bharatheeya coronata L.B. Conc., M.F.O. Marques, J.S. Monteiro,

Gusmao & R.F. Castaneda [MB 819175], p. 358

Corynespora titarpaniensis P. Kushwaha, Raghv. Singh & Sh. Kumar

[MB 818169], p. 272

Dictyotrichocladium Fiuza, Gusmao & R.F. Castaneda

[MB 821638], p. 435

Dictyotrichocladium aquaticum Fiuza, Gusmao & R.F. Castafieda

[MB 821639], p. 435

Elytroderma baikalense T.I. Morozova, Lar.N. Vassilyeva & S.L. Stephenson

[MB 817182], p. 252

Inocybe ahmadii Faroogi, Niazi & Khalid

[MB 817163], p. 259

Lecanora shangrilaensis Z.T. Zhao & L. Lu

[MB 812187], p. 442

Microidium phyllanthi-reticulati Meeboon & S. Takam.

[MB 800332], p. 291

Minimelanolocus aquatilis L.B. Conc., M.F.O. Marques,

Gusmao & R.F. Castafieda [MB 821640], p. 437

Morganella austromontana C.R. Alves, Cortez & R.M. Silveira

[MB 820972], p. 282

Neozygites linanensis X. Zhou & C. Montalva

[MB 814659], p. 310

Pseudobaeospora lilacina X.D. Yu, Ming Zhang & S.Y. Wu

[MB 815567], p. 332

Pseudospiropes piatanensis J.S. Monteiro, Gusmao & R.F. Castafieda

[MB 821604], p. 377

Sporormiella longicolla R.F.R. Melo

[MB 815023], p. 463

Tretoheliocephala Gusmao, J.S. Monteiro & R.F. Castafieda

[[IF 553563], p. 454

Tretoheliocephala compacta Gusmao, J.S. Monteiro & R.F. Castafieda

[IF 553564], p. 454

Xylaria nelumboniformis Hai X. Ma, Lar.N. Vassiljeva & Yu Li

[MB 817858], p. 300

Vul ... MYCOTAXON 132(2)

ERRATA FROM PREVIOUS VOLUMES

VOLUME 131(1)

p. iv, line 33

p. V, line 17

p. V, line 28

p- 88, line 29

p- 93, line 23

p- 153, line 13

FOR: supported by morphological and sequence analyses

READ: supported by new morphological and sequence analyses

FOR: Records of terricolous lichens from Ecuador

READ: Records of terricolous lichens from paramos of southern Ecuador

FOR: Bacidia, Micarea, Sagedia, and Stigmidium spp.

READ: Bacidia, Micarea, Sagedia, and Stigmidium species

FOR: E. sp. [2] READ: E. sp. {2]

FOR: Microbiol READ: Microbiology

FOR: Departamento de Biologia Vegetal IH, Facultad de Farmacia, Universidad

READ: Departamento de Biologia Vegetal I, Facultad de Biologia, Universidad

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-TWO (1)

MYCOTAXON for JANUARY-MARCH 2017 (I-xuI + 1-241)

was issued on April 27, 2017

APRIL-JUNE 2017... IX

FROM THE EDITOR-IN-CHIEF

OUR MYCOLOGICAL RIVER FLOWS ALL TOO SWIFTLY—Several days ago I reflected

over my good fortune in serving 14 years as MycoTaxon Editor-in-Chief. One

enjoyable benefit from editorial supervision is the opportunity to watch fledgling

mycologists progress from first-time authors—most with no clue on how to

formulate a rigorous research paper and lacking the techniques needed to produce

an acceptable manuscript in English—to those who have mastered the basics but

not yet how to publish a crisp valuable taxonomic treatise. Some either never

progress beyond these beginning stages or (more likely) are defeated by the oft-

insurmountable demands of simultaneously juggling teaching assignments, serious

research, ‘grantsmanship, family obligations, and gainful employment in mycology.

However, there are others who do persevere, progressing from ‘top’ student to

newly hired assistant professor, eventually to establish their mycological credentials

as both authors and valued (i.e., careful and helpful) expert reviewers. It is this

last group that we value most and whose career paths we follow closely as they

evolve from mycological neophytes to mycological masters. Eventually personal

correspondence between editor and author develops into friendship (despite the

fact that with increasing age, we editors tend to roar more exasperatedly over

inattention to detail) and intense editorial interest in each author’s achievements.

Alas, mycologists are not immortal, and early 2017 made fatal inroads into the

ranks of our Great Mycologists. Late February brought the news of the unexpected

death of Russian pyrenomycete specialist Larissa VASILYEVA in Vladivostok (two

last papers are published posthumously in this issue with one plate serving as front

cover). In early April we lost hyphomycete giants WALTER GAs in Italy and VApDIM

MEL ’NIK in St. Petersburg, while early June delivered the sad news of the death of

Brazil’s oomycete and zygomycete specialist, JosE IVANILDO DE Souza, in Sao Paulo.

The loss of four such important mycologists warrants more than a single sentence

here. To that end, we include IN REMEMBRANCE (p. xi-xix) on the following pages to

provide photographs and brief biographies of four who gave so much to mycology.

Preparing the text (as always) facing MycotTaxon deadline, I appropriated

material commemorating Mel'nik from Uwe Braun’s thoughtful and warm April

10 publication in SCHLECHTENDALIA. The information is properly attributed, but

I do hope that Uwe will forgive our reprinting one photo here without first asking

permission.

We would like to think that this new section would not be needed again.

Realistically, however, we expect to present such memoria in the future and so

encourage readers and authors to notify us of deaths and send in short biographies

and photographs for us to use as needed.

x ... MYCOTAXON 132(2)

MycoTAXON 132(2) offers 23 research papers by 89 authors representing 13

countries and revised by 40 expert reviewers.

Within its pages are 3 new genera (Arthrotaeniolella, Dictyotrichocladium,

and Tretoheliocephala from Brazil) and 15 species new to science representing

Arthrotaeniolella, Bharatheeya, Dictyotrichocladium, Minimelanolocus, Morganella,

Pseudospiropes, Sporormiella, and Tretoheliocephala from Brazil; Corynespora from

India; Elytroderma from Russia; Inocybe from Pakistan; Microidium from Thailand;

and Neozygites, Pseudobaeospora, and Xylaria from China.

Range extensions and/or new hosts for previously named taxa include new

records of agarics (Gymnopilus in Thailand, Inocybe in Pakistan), lichens (Lecidea in

China), macro- (8 new records in Mexico) and micro-ascomycetes (Cephaliophora

in Poland, Linkosia & Spadicoides in China), stinkhorns (Phallales in Paraguay),

and zygomycetes (Cunninghamella, including a key to the species found in Brazil).

Additional information is presented for the bolete Hourangia cheoi from China and

a full morphological reevaluation of the slime mould genus Cribaria in Mexico

based on scanning electronic microscopy.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

8 July 2017

APRIL-JUNE 2017... XI

IN REMEMBRANCE

[KONRAD] WALTER GAMS_ (1934- me N Y

2017): Dr. Hab. (Prof.) Gams was born :

August 9, 1934, in Zurich, Switzerland,

the son of bryologist and lichenologist

Helmut Gams. He married Sophia

Aaltine Luinge in 1943, was father

to Hedwig and Mechthilde, and died

unexpectedly at age 83 on April 9 in

Bomarzo, Italy, only two weeks after

depositing two Phoma cultures in the

Centraalbureau voor Schimmelcultures

(CBS), renamed Westerdijk Fungal

Biodiversity Institute on 10 February

2017) in Utrecht, Netherlands.

After obtaining his PhD at Austria's

University of Innsbruck in 1960,

Walter joined CBS in 1967 as Senior

Scientist and worked there until

retiring as CBS Director in 1999 and \

as Senior Emeritus Scientist in 2007. Walter Gams stands happily surrounded by

Sometime in the 70s he founded the 20amorph collections in Vienna’s Herbarium W at

the end of the 2005 Nomenclature Section.

famed CBS summer course later

attended by mycologists from all over the world. From 1972 onwards, he also served

as Lecturer at Germany's Technology University in Aachen.

Gams was highly influential in fungal nomenclature, starting with the discussions

surrounding sanctioning of names prior to the 1981 International Botanical Congress

(IBC). Elected to the IBC Permanent Nomenclature Committee for Fungi in 1984,

he succeeded Richard P. Korf as Secretary in 1991 and retired as NCF Secretary in

2005 and from the NCF in 2011. During this time Walter also chaired the MSA

Nomenclature Committee (2001-2004).

A prolific author, with well over 200 publications to his credit, Walter began

work on the zygomycete Mortierella before becoming a world-renowned expert in

asexual (anamorphic) fungi. He teased apart and monographed the difficult speciose

genera of Acremonium, Fusarium, Trichoderma, and Verticillium and participated

in proposing over 33 generic names and 446 species epithets. MycoBank cites

671 names proposed by “W. Gams” and coauthors and lists three genera (Gamsia,

Gamsiella, Gamsylella), 21 species, and one variety named after him.

His more important books include FUNGI IN AGRICULTURAL SOILS (Domsch

& al. 1972), CEPHALOSPORIUM-ARTIGE SCHIMMELPILZE (HYPHOMYCETES) (Gams

1971, with his 1972 defense securing for him the title of “Dr. Hab.), ComPpENDIUM

xl ... MYCOTAXON 132(2)

XVII INTNERNATIONAL BOTANICAL CONGRESS, VIENNA 2005—LeFr: NCF Secretary Gams

discusses why micromycetologists need illustrations as types during IBC Section recess

on July 14. Ricut: On July 16 on the herbarium rooftop, Walter focuses his camera while

Dr. Passauer enthusiastically names the artists atop the Kunst Museum overlooking Maria

Theresa Platz and David Hawksworth walks along the parapet toward the Vienna Woods.

OF Soi, FuNGI (Domsch & al. 1980), SUPPLEMENT AND CORRIGENDUM TO THE

COMPENDIUM OF SOIL FuNGI (Gams 1983), CBS COURSE OF MYCOLOGY (Gams & al.

1987), and THE GENERA OF HYPHOMYCETES (Seifert & al. 2011).

An indefatigable teacher and mentor, Walter also served as Editor for ALLIONIA,

CRYPTOGAMIE-MYCOLOGIE, MyYCOLoGIA AUSTRIACA, Nova HEDWIGIA,

PHYTOPATHOLOGIA MEDITERRANEA, STUDIES IN MyCOLoeGy, and ZEITSCHRIFT FUR

Myxo.oaieE. He published 12 papers in Mycotaxon [including the controversial

“A critical response to the Amsterdam Declaration (2011; 116: 501-512) where he

and 78 other authors passionately defended preservation of dual nomenclature].

Walter also served Mycotaxon frequently (12 times since 2004) as an excellent and

knowledgeable pre-submission expert reviewer.

An honorary member of the British Mycological Society and Mycological Society

of America (MSA), Walter received MSA’s highest award, Distinguished Mycologist,

in 2005 and the Anton de Bary medal from the Deutschen Phytomedizinischen

Gesellschaft in 2012. One letter nominating Dr. Gams as Distinguished Mycologist

noted “Five personal characters and abilities mark a person as being distinguished

and a cut above others. These are: passion for the work; excellence in its execution;

creativity in approach; communication of research results; and service to science”

(2006, INocuLUM 56(5): 15). Walter epitomized all five.

I was fortunate to meet Walter personally over a glass of wine at the 1994 IMC7

in Vancouver BC. Knowing of my nerdish interest in fungal nomenclature, he invited

me to attend a meeting of the IBC NCF the next morning. I was hooked sufficiently

that he nominated me for membership on the committee, to which I was elected

at the 1999 St. Louis IBC. Probably because I submitted opinions (well thought

out or otherwise) on everything before his ballot deadlines, he later mentored my

taking over as Secretary at the 2005 Vienna IBC Nomenclature Section, where he

APRIL-JUNE 2017... XII

Lorelei Norvell

Courtesy of Walter Gams

eh2

sii

Lerr— Walter and David Hawksworth discuss changes in the Vienna Code over dessert

and wine at the IBC XVII in Vienna. RichHt— MSA Distinguished Mycologist Gams would

soon be soaked by boiling surf on the black sands of the ‘Big Island’ (Hawai’i) in 2006.

BELow — Walter stands on a summit in the Italian alps with Takamichi Orihara in June 2012.

and David Hawksworth maneuvered me through the sessions and Viennese streets

and restaurants. While nomenclatural differences between opposing factions

run deep (often vituperatively so), I was fortunate to witness first hand the rather

stunning collegial amiability evinced by those two mycological nomenclatural giants

at lunches and dinners, each glancing at me with twinkling eyes to see whether I ‘got’

a particular point contrary to the other's point of view.

After Vienna, Walter became a good friend and frequent correspondent. We

met frequently at MSA, IMC, and IBC meetings and exchanged monthly Emails

and photos. All of us lucky enough to receive his annual December PDFs became

an integral and appreciative member of his far-flung mycological family. It may

seem odd to regard death at 83 as being cut down in one’s prime, but given Walter’s

peripatetic nature and innate vigor, he was. We will all miss him.

ResearchGate photo

XIV ... MYCOTAXON 132(2)

VADIM ALEXANDROVICH MEL’NIK (1937-2017):

Note: The information below (and more) comes mainly

from Uwe Braun's excellent personal tribute to his close

friend and colleague (Schlechtendalia 32: 29-34, 2017).

Vadim Mel'nik was born in Daugavpils, Latvia, on

March 16, 1937. After researching spruce canker in

Kaliningrad forests, he graduated in 1960 as ‘forestry

engineer’ from the Leningrad Forestry Academy and

began post-graduate work at the Komarov Botanical

Institute in 1962, where he obtained his PhD based

on phytopathogenic asexual fungi of the Leningrad region and in 1986 received

his final academic credentials in his defense of his research on coelomycetes of the

former USSR. At the Komarov, Mel'nik advanced to Senior Research Fellow in 1977,

was promoted to Leading Researcher (Laboratory of Systematics and Geography

of Fungi) in 1998, and served as Curator of the fungal herbarium LE for 15 years.

Throughout his career, Mel'nik collected widely throughout the former Soviet

Union. A highly esteemed hyphomycete and coelomycete specialist, he also identified

“countless fungal samples collected by colleagues from all over the world” and was

frequently invited to speak at symposia and important mycological meetings in

other countries. Toward the end of his life, he concentrated on microfungi collected

during a research program by the Vietnam-Russian Tropical Research & Technology

Centre for which he published numerous papers and several new species.

Among more than 220 scientific publications, his most important publications

include his 1977 Ascochyta monograph and its translation, KEY TO THE FUNGI OF

THE GENUS ASCOCHYTA LIB. (COELOMYCETES) (Melnik 2000), IMPERFECT FUNGI

ON SPECIES OF TREES AND SHRUBS (Mel'nik & Popushoi 1992), CERCOSPOROID

FUNGI FROM RUSSIA AND ADJACENT COUNTRIES (Braun & Melnik 1997), and two

volumes covering the coelomycetes and dematiaceous hyphomycetes of Russia

(Melnik 1997, 2000). ResearchGate (https://www.researchgate.net/profile/Vadim_

Melnik) provides additional information on 52 Melnik research publications.

Melnik was also a “gifted editor and proofreader” who reviewed manuscripts for

numerous mycological journals (including Mycotaxon) and served on the editorial

boards of FUNGAL DIVERSITY, FUNGAL DIVERSITY RESEARCH SERIES, MYCOLOGIA

BALCANICA/MycosioTa, and MycospHere. He supported MIKOLOGIYA I

FITOPATOLOGIYA from its inception in 1967 and also served as Editor or Editor-in-

Chief for numerous Russian book projects and series.

Only a few Russian mycologists have received the Russian A.A. Yachevky medal

awarded to Vadim Mel'nik in 2012. Two genera (Melnikomyces, Melnikia) and

four species have been named in his honor, while he introduced “123 taxonomic

novelties, most of them new species, including three new families and three new

genera’ (Braun 2017; MycoBank lists ~191 taxonomic names).

APRIL-JUNE 2017... XV

printed from Braun (2017)

VADIM MEL’NIK surrounded by pointsettias in Shangxi, China in 2014

Prof. Braun concludes his tribute with: “A long-standing friendship, over decades,

defying all kinds of social and political changes, has been suddenly interrupted.

I will painfully miss him, his unselfish generosity, always ready to help, his

open-minded character, and his regular phone calls.... If there would be a ‘Walk

of Fame’ for mycologists, he undoubtedly deserved his own star.... He will be

remembered by all as not only a gifted mycologist but also someone who was always

prepared to help and train others. Vadim Mel'nik lost his wife early to cancer. He is

survived by his daughter, Irina, and grandchildren.”

Dr. José IVANILDO DE SOUZA [1973-2017]: We received the sad news of the death

at the age of 44 of this irreplaceable young mycologist only three weeks ago. Dr. José

Ivanildo graduated in the biological sciences from Sao Judas Tadeu University in

1995, where he focused on the taxonomy, ecology, and phylogeny of fungi (Mucorales)

and oomycetes (Oomycota). He received his masters (2001) and PhD (2006) from

Paulista State University Julio de Mesquita Filho (UNESP) and served his post-

doctorate at the Center for Parasitological and Vector Studies (CONICET, UNLP)

in Argentina in 2011. At the time of his death José Ivanildo was Scientific Researcher,

Director of Technical Services in Nuclear Research in Mycology, at the Instituto de

Botanica in Sao Paulo.

XVI ... MYCOTAXON 132(2)

Courtesy of André Cabral Santiago —

JOSE IVANILDO DE SOUZA at a forest overlook in Brazil in 2014

“J.L. Souza” is cited in MycoBank as author or coauthor of five taxonomic names. He

was a former editor of CHECK LisT and BRAZILIAN JOURNAL OF BOTANY and served

as expert peer reviewer for MycoLocy, Mycotaxon, SYDOWIA, BRAZILIAN JOURNAL

OF Botany, HOEHNEA, and Nova Quimica. As senior author for MycoTAxoNn he

published a 2007 paper on selected mucoralean fungi from soils contaminated with

toxic metals and “Zygomycetes from the Reserva Bioldgica de Mogi Guau” (2011).

A list of José Ivanildo’s 29 research publications and other mycological contributions

may be accessed on (https://www.researchgate.net/profile/Jose_De_Souza). Those

who understand Portuguese will enjoy his “Get to know all about fungi’ interview

(https://www.youtube.com/watch¢v=8UZzPMGWdvI) recorded by Todo Seu in 2013.

Dr. André Cabral Santiago (Federal University of Pernambuco), who shared the

above photograph of his colleague, wrote: “Dr. José Ivanildo de Souza [1973-2017]

was a renowned Brazilian mycologist who dedicated his professional life to the

study of zygosporic fungi, contributing greatly to the knowledge of the mucoralean

diversity in Brazil’s Atlantic Forest and Cerrado regions in Brazil. In recent years

he published several important manuscripts on zoosporic fungi. He worked as a

researcher at the Institute of Botany of Sao Paulo and died as a consequence of

a pancreatic cancer on June 5, 2017. During the time he worked as a mycologist,

Dr. José Ivanildo inspired several mycologists to follow in his footsteps. He was

a professional who always showed humility, ethics, and professionalism in his

relationship with other researchers and students.’

Haixia Wu

PAA Waa

APRIL-JUNE 2017... XVII

Larissa NIKOLAEVNA VASILYEVA [1950-2017]. Larissa Vasilyeva (also transliterated as

Larisa and Vasiljeva, Vassiljeva, or Vassilieva based on different linguistic conventions)

was born in Kursk, western Russia. After graduating from Leningrad State University

in 1972, she moved to Vladivostok, where she mastered pyrenomycetes while at the Far

East Branch of the Russian Academy of Sciences Institute of Biology and Soil Science.

Her PhD dissertation, PYRENOMYCETES OF THE RUSSIAN FAR EAST (1992), established

her as an expert in the field, and MycoBank cites ~458 taxa (not including the two

new species published in this issue) authored or coauthored by “Lar.N. Vassiljeva’.

In addition to her many pyrenomycete works, Dr. Vasilyeva wrote several philosophical

treatises on general taxonomic theory—SysTEMATICS IN MycoLoGcy (1990), PLATONISM

IN SYSTEMATICS (1992)—and coauthored the popular MACROFUNGI ASSOCIATED WITH

OAKS OF EASTERN NorTH AMERICA (2008).

Dr. Vasilyeva was active throughout Asia and North America, serving several

terms on the IMA Asian Mycology Committee and as member of the Mycological

Society of Japan, Mycological Society of America, and the Russian Academy of

Sciences. Her memorial in the Big THICKET BULLETIN notes that Larissa “loved

to collect fungi, wandering through the woods humming softly to herself. She

specialized in very small ascomycetes ... partly, perhaps, because she had myopia or

near-sightedness that allowed her to see clearly the small black dots of pyrenomycetes

scattered along a dead branch. She would pick up a likely substrate, pull off her thick-

lensed glasses and examine the stick without a hand lens because, basically, she had

a “built-in” hand lens for eyes. In addition, she had a sense of the biology of these

=2n

LarIssA VASILYEVA on Changbaoi Mountain (Jilin Province, northern China) in 2013

Xvull ... MYCOTAXON 132(2)

LarISssA VASILYEVA in tropical Hainan Province (southern China) in 2013

fungi and thus knew where to look for them. She was fearless in her ability to endure

hot and cold weather in search of her beloved pyrenomycetes.” (Accessed 7 July 2017:

http://www.thicketofdiversity.org/blog/tod-researcher-larissa-vassiljeva/)

Dr. Steve Stephenson (University of Arkansas) frequently collaborated with

Larissa during her exploration of the “Asa Gray disjunction” that demonstrated that

pyrenomycetes also produce closely related organisms in eastern North America and

eastern Asia, but nowhere else in the world. He noted Vasilyevas observation, “In

some cases, morphologically similar fungi present in both regions and considered

to represent the same species may instead be two distinct species. In other cases, a

single species not recognized as such may occur in both regions”

My own encounters with this formidable scholar with the gentle soul began

in 2000 with Larissa’ desire to publish ‘Further comments on the ‘PhyloCode’

(INOCULUM 51(6): 5-7) to add to an earlier Scott Redhead commentary (INOCULUM

51(3): 1-3). She concluded, “In general, I do not think that such word combinations

as ‘phylogenetic nomenclature’ or ‘PhyloCode’ are well chosen. If taxonomists are

to be able to subordinate characters properly and ... to circumscribe more natural

groups, the taxon names (nomenclature) will automatically be ‘phylogenetic. As for

the Code, it calls for the ordering of taxonomists, not taxa. If two taxonomists in

Europe and Asia describe the same taxon, there really should be rules coordinating

what that taxon can be called”

Haixia Wu

APRIL-JUNE 2017... XIX

We exchanged several Emails while I ‘polished’ her English (my Russian training

coming to our rescue) and simplified her complex thoughts on taxonomy and

nomenclature for MSA readers. Our amiable electronic friendship resumed in

I 2004 with Mycotaxon, for which she contributed to 19

papers (including two published posthumously in 132-2)

and thoroughly reviewed 11 others. We met all too briefly

at Edinburgh’s 2010 IMC, memorialized by the blurry

»— photo at left. She tactfully returned the much lovelier

Ss one on p. xviii, taken by Haixia Ma in 2013. Although we

| had both been diagnosed with the same cancer, Larissa’s

2016 therapy failed her, and she left us far too soon on

® February 23, only one week after her 67" birthday.

MY COTAXON

April-June 2017— Volume 132, pp. 243-250

https://doi.org/10.5248/132.243

Three species of Linkosia and Spadicoides new to China

ZHAO-HuAN Xu’, D1iAN-MING Hv’, YOU-QIANG Luo’ & JIAN Ma’*

‘College of Agronomy & *College of Bioscience and Bioengineering,

Jiangxi Agricultural University, Nanchang, 330045, China

*CORRESPONDENCE TO: majian821210@163.com; jxaumj@126.com

ABSTRACT—Specimens of Linkosia longirostrata, Spadicoides klotzschii, and S. sylvatica were

collected on dead branches of unidentified plants and identified as new records from China.

They are described and illustrated based on the Chinese material. Keys are also provided to

Linkosia and Spadicoides species recorded from China.

KEY wWoRDS—asexual morph, hyphomycetes, taxonomy

Introduction

Linkosia A. Hern.-Gut. & B. Sutton, established in 1997 with Sporidesmium

coccothrinacis A. Hern.-Gut. & J. Mena [= L. coccothrinacis (A. Hern.-Gut. &

J. Mena) A. Hern.-Gut. & B. Sutton] as type species, is mainly characterized by

conidiophores that are reduced to monoblastic conidiogenous cells producing

solitary, distoseptate conidia (Hernandez-Gutiérrez & Sutton 1997, Wu &

Zhuang 2005). After Zhang et al. (2009) provided a key to the then-known six

species, six additional species were described by Ma et al. (2011), Almeida et

al. (2014), Delgado (2014), and Conceicao et al. (2016). Of the twelve species

currently accepted in the genus (Conceicao et al. 2016), five have been recorded

from China (Wu & Zhuang 2005, Zhang et al. 2009, Ma et al. 2011).

Spadicoides S. Hughes, established in 1958 with Helminthosporium binum

Corda [= S. bina (Corda) S. Hughes] as type species, is diagnosed by distinct,

unbranched or sparingly branched conidiophores with integrated, polytretic,

terminal or intercalary conidiogenous cells producing solitary, acropleurogenus,

unicellular or euseptate conidia (Hughes 1958, Ellis 1971, Sinclair et al. 1985,

Seifert et al. 2011). Forty-two species have been described, of which fifteen have

244 ... Xu &al.

been recorded from China (Zhou et al. 1999, Lu et al. 2000, Ho et al. 2002,

Wong et al. 2002, Cai et al. 2004, Li et al. 2010, Ma et al. 2010, 2012, 2014,

2016, Xia et al. 2014). Goh & Hyde (1996) and Ma et al. (2016) have provided

synoptic tables of relevant morphological characters of Spadicoides species.

During our continuing surveys of saprobic microfungi from the forests of

southern China, we have collected specimens representing three new Chinese

records of Linkosia and Spadicoides on dead branches. They are described and

illustrated below. The specimens are conserved in the Herbarium of the Department

of Plant Pathology, Jiangxi Agricultural University, Nanchang, China (HJAUP).

Linkosia longirostrata G. Delgado, Mycotaxon 129(1): 42, 2014. FIG. 1

CONIDIOPHORES absent or very reduced, erect, 1-septate, dark brown,

21-35 um long, 6-12.5 um wide at the swollen base. CONIDIOGENOUS CELLS

monoblastic, integrated, terminal, lageniform or subcylindrical, smooth,

brown to dark brown, determinate or occasionally percurrent, 15-27 x 5-6 um,

3-4 um wide at the truncate apex. Conrp1A holoblastic, solitary, acrogenous,

A B D

[ i E 3 ee

Fic. 1. Linkosia longirostrata (HJAUP M0162). A, B. Conidiophores, conidiogenous cells and

conidia; C. Conidiophores and conidiogenous cells; D, E. Conidia. Scale bars = 20 um.

Linkosia and Spadicoides spp. new for China... 245

straight or curved, obclavate to obclavate-rostrate, pale brown, smooth,

10-18-distoseptate, 82-126 um long, 6.5-8.5 um wide in the broadest part,

3-3.5 um wide at the truncate base, apex extended into a pale brown to brown

rostrum, 2-2.5 um wide, and invested in 1-3 spherical or subspherical, hyaline

or subhyaline mucous tunicae ca 6-15 um diam.

SPECIMEN EXAMINED: CHINA, JIANGXI PROVINCE, Lushan Mountain, on dead

branches of an unidentified broadleaf tree, 7 November 2014, J. Ma (HJAUP M0162).

CoMMENTS - Delgado (2014) recorded this fungus on rachides of dead leaves

from the palm species Acoelorrhaphe wrightii in Florida, USA, and compared it

with similar species, including Linkosia coccothrinacis, L. ponapensis (Matsush.)

R.F. Castaneda et al., L. refugia (B. Sutton & Pascoe) D.A.C. Almeida & Gusmao,

and L. canescens (B. Sutton & Pascoe) D.A.C. Almeida & Gusmao. This is the

first report of L. longirostrata in China. Our specimen is similar to the holotype

from USA, but the holotype has smooth or slightly verrucose conidiogenous

cells, and finely rough conidia without mucous tunicae at the apex (Delgado

2014); also, our specimen was collected from dicotyledenous wood, whereas

the holotype was collected from monocotyledonous rachides.

Key to species of Linkosia from China

1. Conidia with 27-30 distosepta, 300-380 x 12-14um ............... L. multisepta

t. Conidia with <21 distosepta and: <210- par long 22. oye ese ew when on ofee opti oie s v)

2. Gonidia IG niin-on dd S=4-5 fe WAS ey oe HPAES oR oleh oh hel a. oe alae 3

BE Glelaital cao Dg Ui eng Atel amr read Mies Peas Sets Senha cn ene ACE SSR ARE re Ree eA 4

3. Conidia with mucilaginous sheath at the apex, 100-160 x 13-15 um .. L. fusiformis

3. Conidia lacking mucilaginous sheath, 122-177 x 12-16 um .............. L. mori

4, Conidia with 16-21 distosepta, 160-210 x 7.5-9.5 um .............006- L. hibisci

AO OmidiavcAd 6PM ON Ge he fae S ne NS ae ot oe oh he Ao Gl cok Teh 5

5. Conidia with 12-14 distosepta, 110-125 x 9-12 um ................. L. obclavata

5. Conidia with 10-18 distosepta, 82-126 x 6.5-8.5 um ............. L. longirostrata

Spadicoides klotzschii S. Hughes, Fungi Canadenses: no. 8, 1973. FIG. 2

CONIDIOPHORES macronematous, mononematous, unbranched, erect,

straight or flexuous, cylindrical, smooth, brown to dark brown, 5-8-septate,

85-135 x 3.5-4.5 um. CONIDIOGENOUS CELLS polytretic, integrated, terminal

and intercalary, cylindrical, smooth, brown, 6-11.5 x 2.5-3.5 um. CONIDIA

acropleurogenous, solitary, dry, obovoid, smooth, brown to dark brown, basal

cell paler, 2-euseptate, 10-11.5 um long, 5-6 um wide in the broadest part,

0.5-1 um wide at the base.

246 ... Xu & al.

| | oe

20m

Fic. 2. Spadicoides klotzschii (HJAUP M0037). A-C. Conidiophores, conidiogenous cells and

conidia; D. Conidiophores showing conidiogenous pores; E. Conidia. Scale bars = 20 um.

SPECIMEN EXAMINED: CHINA, GuiIzHOU PROVINCE, Leigongshan National Nature

Reserve, on dead branches of an unidentified broadleaf tree, 7 September 2013, J. Ma

(HJAUP M0037).

ComMENnTs -— Hughes (1973), who reported Spadicoides klotzschii on dead wood

of Alnus and from unidentified hosts in British Columbia, Canada, compared

it with S. obovata (Cooke & Ellis) S. Hughes. Subsequently, Pinruan et al.

(2007) recorded this fungus from Thailand. Compared with our specimen, the

holotype has slightly narrower conidia (4-5.5 um) and longer conidiophores

(55-220 um; Hughes 1973). This is the first report of S. klotzschii in China.

Spadicoides sylvatica Heredia, R.F. Castafieda & R.M. Arias,

Mycotaxon 130(2): 512, 2015. Fien3

CONIDIOPHORES macronematous, mononematous, unbranched, erect,

straight or flexuous, cylindrical, determinate or occasionally percurrent,

Linkosia and Spadicoides spp. new for China ... 247

A B C

Fic. 3. Spadicoides sylvatica (HJAUP M0112). A-C. Conidiophores, conidiogenous cells and

conidia; D. Conidiophore apex showing conidiogenous pores; E. Conidia. Scale bars = 20 um.

smooth, dark brown, 6-13-septate, 130-220 x 4-7 um. CONIDIOGENOUS

CELLS polytretic, integrated, terminal and intercalary, cylindrical, smooth,

brown, 14-25 x 3.5-5 um. Conipia acropleurogenous, solitary, dry, ovate to

obclavate, gradually tapered to an obtuse apex, smooth, brown, apical cell paler,

(3-)4-euseptate, 25.5-35.5 x 6.5-9.5 um, 1.5-2.5 um wide at the base, 1.5-2.5

um wide near apex.

SPECIMENS EXAMINED: CHINA, GUANGDONG PROVINCE, Liuxihe National Forest

Park, on dead branches of an unidentified broadleaf tree, 10 July 2014, J. Ma (HJAUP

M0112, HJAUP M0116).

ComMENTS - Spadicoides sylvatica was described from Mexico and compared

with S. yunnanensis L.G. Ma & X.G. Zhang (Heredia et al. 2015); it has not

previously been recorded from China. Our collection corresponds well with the

original description of S. sylvatica except that the holotype has slightly smaller

conidia (22-32 x 7-8 um) with verruculose apical cells (Heredia et al. 2015).

248 ... Xu & al.

Key to species of Spadicoides from China

he CORI ANON SE PLALes 2 ued. wo gh se eae ane ug DE ease bie eis bees be 2

ZO Clia Se Piet Peo Mla rc Neo Se on gta aco onghy e's mip ans malt ons gy eg A nga pcp eo -:

2. Producing a synanamorph predominantly with 2-septate synconidia .. S. wufengensis

2y Lacking a-sy manana, bai tarhcengerhec tad aktie Badnina Rade biade: oeeBale: hed s S

SiC Gnidia smoot, 46,5265 4 iii x ye le pone Lay Ce hoe Ce Bee Cael ee S. atra

3. Conidia-verruculose, 11-18 X 4-650) Asc ee dence ee ced smnew en ee es S. arengae

4; Gonidia-withibotheusepta-and.-distosepta: «.. c.sc seca peta te erect at erect ah ered athe 5

4 Comidiawithonly: CUSepid” ak iy ae he ral tA ee a Gee § 6

5. Conidia either 0-1-euseptate, or with 1 euseptum + 1 distoseptum,

ONS G DOB: MIT sate cr peietea pe NP ie St gs SNA pr ae go PRE ge PERS orto S. hodgkissii

5. Conidia with 2 eusepta + 1 distoseptum,

US STA SSO, iP gg Mga he leet pier to SMe t ori dak? ore tush ork S. versiseptatis

ORC OHICa VeTEUCTLOS Sat sc. tata. on Meanie tea aot oan ee tan ween ReBes ss hetbess Paeers he Boe sn 7

GeO onidia smooth eestees veh aes yearns whee einge a eae a ears nace re ey eet oe 8

7. Conidia 5—7-euseptate, 30-72.5 x 5-7.5 UM ....... eee eee eee S. bambusicola

7. Conidia 9-12-euseptate, 98-166 x 8.5-12 um.............00-. S. bawanglingensis

BAS OIC Ey WET SNCONOT OMG. a pivag.o meee. s glen eatin Saag ¢-m te mente eee eae te opaete net 9

SUC OMIA -CONCOLTOMS: SAitst s oliae a IMAC on RA ates Reta Eee st gS 6 on ati Re ae 12

9. Conidia 3—4-euseptate, 25.5-35.5 x 6.5-9.5UM 1... eee eee eee eee S. sylvatica

9. Gonidiaspredominantly 2-euseptate v1 oxy. ducn en ek ete eines De rie Mee eens 10

10. Conidia: 10=tel 53 SSG lit, cdc ats a peat gnpea’epadts meta a-es eat S. klotzschii

LOMC oniciia at Least) 5 tO Th ote on pias nekecis nha s neha oa mages acagheeacagtene te 11

LikConicdiaveveidsl5-22 36-710 s ti oe ee et ol eth oe ok alee Wats S. camelliae

11. Conidia obovate, clavate, 12.5-16 x 6-8.5UM........... eee eee eee S. obovata

12. Conidia bacilliform, 3-euseptate, 33-55 x 5.5-6.5 um ............ S. bacilliformis

12. Conidia obclavate, oval to reniform or obpyriform to ovoid ................... 3

13. Conidia 11-15-euseptate, 65-115 x 11.5-13 um ................ S. multiseptata

13; Comidia <8-euseptateand S65-t On ee ha oi hie eG ah a noes nats 14

14, Conidia 2-3-euseptate, <28 um long, rounded or acute at the apex ........... lhe

14. Conidia at least 5-euseptate, 40-65 um long, rostrate .................. 0008. 16

15. Conidia oval to reniform, 16-23 x 6.5-8 UM ......... 2. eee eee S. longchiensis

15. Conidia obpyriform to ovoid, 18.5-28 x 6.5-10 um.............. S. yunnanensis

16. Conidia 6-8-euseptate, 40-65 x 4.5-5.5 UM ........ eee eee ee eee S. lushanensis

16. Conidia 5(-6)-euseptate, 40-65 x 10.5-12.5 um... 2... eee eee S. rostrata

Acknowledgments

The authors express gratitude to Dr. Rafael F. Castafteda-Ruiz and Dr. Li-Guo Ma

for serving as pre-submission reviewers and to Dr. Shaun Pennycook for nomenclatural

Linkosia and Spadicoides spp. new for China ... 249

review and Dr. Lorelei L. Norvell for editorial review. This project was supported by the

National Natural Science Foundation of China (Nos. 31360011, 31460009, 31500021)

and the Education Department of Jiangxi Province of China (No. GJJ160357).

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MY COTAXON

April-June 2017— Volume 132, pp. 251-255

https://doi.org/10.5248/132.251

Ascomycetous fungi of Siberia. 3.

Elytroderma baikalense sp. nov.

LARISSA N. VASILYEVA'|t], TATYANA I. MOROZOVA”?

& STEVEN L. STEPHENSON*

"Institute of Biology & Soil Science,

Far East Branch of the Russian Academy of Sciences, Vladivostok 690022, Russia

*Siberian Institute of Plant Physiology and Biochemistry,

Siberian Branch of the Russian Academy of Sciences, Irkutsk 664033, Russia

*Irkutsk Interregional Veterinary Laboratory, Irkutsk 664005, Russia

‘Department of Biological Sciences, University of Arkansas,

Fayetteville, Arkansas 72701, USA

“CORRESPONDENCE TO: slstephenson@uark.edu

ABSTRACT—Elytroderma baikalense is described herein as a species new to science, based on

material collected from dying needles of Pinus sylvestris in the Lake Baikal region of Siberia.

Images of ascomata, asci and ascospores are provided.

Key worps—Ascomycota, pine disease, Rhytismataceae, taxonomy

Introduction

This paper is the third report on the ascomycetous fungi from the Lake Baikal

region of Siberia (Morozova & Vasilyeva 2003; Vasilyeva & Morozova 2004).

In 1995, a species of Elytroderma was found on second-year needles of Pinus

sylvestris L. on Olkhon Island in Pribaikalsky National Park, Lake Baikal, and

identified as E. deformans (Weir) Darker (Morozova 1996). However, additional

study indicated that Elytroderma deformans occurs in North America, mostly on

members of Pinus sect. Trifoliae (American hard pines, including P attenuata,

P. banksiana, P. contorta, P. echinata, P. jeffreyi, and P. ponderosa), but also on

members of Pinus sect. Parrya (P. cembroides and P. edulis) (Hiratsuka 1987,

Stone 1997). The range of E. deformans encompasses a major part of Canada

252 ... Vasilyevat, Morozova & Stephenson

(British Columbia, the Northwest Territories, and Ontario) as well as the

western United States (Arizona, California, Colorado, Idaho, Montana, Oregon,

and Washington) (Minter 1980).

A second species, Elytroderma torres-juanii Diam. & Minter, was described

on Pinus brutia Ten. from Greece; earlier names based on Spanish material—

“Hypoderma hispanicum’ J. Torres (Martinez & Torres 1965) and “Elytroderma

hispanicum” (J. Torres) Darker (Darker 1967)—were invalid because no type

specimen was indicated (Diamandis & Minter 1979). Later, E. torres-juanii was

found on Pinus stankewiczii [= P. brutia var. pityusa| and P. thunbergii in the

Crimea (Minter & Dudka 1996), and its overall distribution was characterized as

“known on various species of pines from parts of Europe with a Mediterranean

climate” (Minter: http://www.cybertruffle.org.uk/papers/rhytukra/elytrode.htm).

The European pines recorded as hosts of Elytroderma torres-juanii are in

Pinus sect. Pinus, together with P. sylvestris, the host of the fungus from the Lake

Baikal region. Thus, E. torres-juanii and the Siberian Elytroderma have hosts

from the same section of Pinus. However, from a biogeographical standpoint,

E. deformans would seem to be more likely to occur in eastern Siberia, with

a distribution similar to that of Delphinella balsameae (Waterman) E. Mill.

which has been reported from northeastern North America and also found

in the Irkutsk region (Morozova & Vasilyeva 2003). This distribution does

not apply to the Siberian Elytroderma, which differs from the two previously

described species in ascospore characters.

Materials & methods

The specimens considered herein were collected by T.I. Morozova on Olkhon Island

in Lake Baikal in 1995 and 1998. The specimens were studied microscopically using

standard techniques. Images of the ascomata were obtained by T. I. Morozova using

a Stereo Discovery V12 microscope equipped with an AxioCam ERc 5s camera, and

images of asci and ascospores were obtained with an AxioLab Al microscope and the

same type of camera.

Taxonomy

Elytroderma baikalense T.1. Morozova, Lar.N. Vassilyeva & S.L. Stephenson,

sp. Nov. FIGS. 1-3

MycoBank MB 817182

Differs from Elytroderma deformans by its smaller ascospores and the absence of

witches’ broom symptoms and from E. torres-juanii by its smaller hyaline ascospores.

Type: Russia, Irkutsk region, Lake Baikal, Pribaikalsky National Park, Olkhon Island,

Olkhonsky District, 15 km east of the village of Khuzhir, on dying needles of Pinus

sylvestris, 27 July 1998, T.I. Morozova (Holotype, VLA P-2934; isotype, IRK 45988).

Elytroderma baikalense sp. nov. (Russia) ... 253

Fics. 1-3. Elytroderma baikalense (holotype, VLA P-2934). 1. Ascomata. 2. Ascus

with ascospores. 3. Ascospores. Scale bars: 1 = 3.0 mm; 2 = 20 um; 3 = 25 um.

254 ... Vasilyevat, Morozova & Stephenson

Erymo.oey: The specific epithet refers to Lake Baikal.

ASCOMATA embedded in brown needles or in brown portions of otherwise

green needles, scattered or confluent when arranged in 2-3 rows along the

needle, appearing as thin black lines, ranging from 2-3 to 15-20 mm long;

forming diffuse brown zone lines across the needle; when viewed in a mid-

point vertical section, the hymenium is covered by a clypeus comprising

blackened fungal cells. Asci unitunicate, mostly saccate, 150-180 x 30-40 um,

with a J-negative apex, containing eight ascospores arranged in a fasciculate

fashion. AscosporeEs clavate, hyaline, 1-septate, smooth, 60-80 x 6-8 um,

enveloped in a thin gelatinous sheath approximately 2 um thick.

ADDITIONAL SPECIMENS EXAMINED: RUSSIA, IRKUTSK REGION, Lake Baikal,

Pribaikalsky National Park, Olkhon Island, Tashkiney Valley, on dying needles of Pinus

sylvestris, 19 July 1995, T.I. Morozova (IRK 45989); 15 km E of the village of Khuzhir, on

dying needles of Pinus sylvestris, 27 August 1998, T.I. Morozova (IRK 46015); Sarayskaya

Bay, on dying needles of Pinus sylvestris, 17 June 1997, T.I. Morozova (IRK 45990).

Notes: The two previously described Elytroderma species produce larger

ascospores than E. baikalense: 90-130 x 8-10 um for E. deformans and 130-165

x 9-12 um for E. torres-juanii. Elytroderma deformans is known to cause a

witches’ broom of pine, but this has not been observed in either E. baikalense

or E. torres-juanii. The ascospores of E. torres-juanii also differ from those of

E. baikalense by their faintly yellowish brown coloration (Diamandis & Minter

1979).

Discussion

Needle viability in Pinus sylvestris is observed to be the longest on Olkhon

Island for the entire region where P. sylvestris occurs. The island is the only

place where needles remain viable on the trees for 10-12 years. Such extended

viability is more characteristic of dark coniferous trees (e.g., Abies sibirica

Ledeb. or Picea abies (L.) H. Karst.). The normal age of Pinus sylvestris needles

is 4—5 years in ecologically pristine places around Lake Baikal, but this drops to

only 2-3 years in the industrial centers and cities (Morozova 2004).

Three hotbeds of this newly described pine needle disease were found along

the southeastern coast of Olkhon Island, with the distance between them

being about 10-20 km. Over 20 years, these hotbeds did not spread but rather

remained discrete. The affected trees lose as much as 75% of their foliage, and

in the crown of the tree needles remain for only 1-2 years. However, the disease

severity fluctuates from year to year. Sometimes, only individual needles are

involved, but there are other years when all of the second-year needles become

yellow and fall. The ascomata mature in August.

Elytroderma baikalense sp. nov. (Russia) ... 255

Although the trees in the hotbeds appear to be greatly weakened, we do

not recommend cutting them. The older trees mostly have affected needles

on the lower branches, which does not lead to a condition of complete blight.

The young affected trees fall naturally and die out. Constant monitoring of the

disease is recommended, and young plants should not be transported from

Olkhon Island.

Acknowledgments

The research reported herein was funded in part by a grant (14-44-040307 Siberia)

from the Russian Foundation of Fundamental Studies. We are thankful to Donald

Pfister (Harvard University, Cambridge, USA) and Wen-Ying Zhuang (Institute of

Microbiology, Chinese Academy of Sciences, Beijing) for the reading of our manuscript

and their valuable comments.

Literature cited

Darker GD. 1967. A revision of the genera of the Hypodermataceae. Canadian Journal of Botany 45:

1399-1444. http://dx.doi.org/10.1139/b67-145

Diamandis S, Minter DW. 1979. Elytroderma torres-juanii sp. nov. from Greece. Transactions of the

British Mycological Society 72(1): 169-172. http://dx.doi.org/10.1016/S0007-1536(79)80025-X

Hiratsuka Y. 1987. Forest tree diseases of the Prairie Provinces. Canadian Forestry Service,

Northern Forestry Centre, Edmonton, Canada.

Martinez JB, Torres Juan J. 1965. Enfermedades de las coniferas espafolas. Boletin—Instituto

Forestal de Investigaciones y Experiencias 36(88). 95 p.

Minter DW. 1980. Elytroderma deformans. CMI descriptions of pathogenic fungi and bacteria

No. 655. Commonwealth Agricultural Bureaux, Kew.

Minter DW, Dudka IO. 1996. Fungi of Ukraine: a preliminary checklist. IMI, Egham & M.G.

Kholodny Institute of Botany, Kiev.

Morozova TI. 1996. Fungal diseases of the forest in the Pribaikalsky National Park. 124-126, in:

The conservation of ecosystems and the organization of monitoring of specially protected

territories. Irkutsk University Press, Irkutsk (in Russian).

Morozova TI. 2004. Fungal diseases of Pinus sylvestris L. in Baikal Siberia. 183-184, in: Structural

and functional organization and dynamics of forests. V.N. Sukachev’s Institute of Forests,

Krasnoyarsk (in Russian).

Morozova TI, Vasilyeva LN. 2003. Ascomycetous fungi of Siberia. I. Delphinella balsameae—

the causal agent of the shoot blight of Siberian fir. Mikologiya i Fitopatologiya 37(1): 59-61

(in Russian).

Stone J. 1997. Other foliage diseases of pines. 59-61, in: EM Hansen, KJ Lewis (eds). Compendium

of conifer diseases. APS Press. St. Paul.

Vasilyeva LN, Morozova TI. 2004. Siberian Ascomycetes. II. Species of the genus Lophodermium on

Pinus spp. Mikologiya i Fitopatologiya 38(5): 42-47 (in Russian).

MY COTAXON

April-June 2017— Volume 132, pp. 257-269

https://doi.org/10.5248/132.257

Inocybe ahmadii sp. nov. and

anew record of I. leptocystis from Pakistan

AYESHA Farooat’*, FAuzIA AQDUS’, ABDUL REHMAN NIAZ1I’,

SANA JABEEN” & ABDUL NASIR KHALID'

' Department of Botany, University of the Punjab,

Quaid-e-Azam Campus-54590, Lahore, Pakistan

? University of Education, Faisalabad Campus,

Samanabad, Faisalabad, Pakistan

*CORRESPONDENCE TO: ayeshafaroogi27@gmail.com

ABSTRACT—Two smooth-spored Inocybe species, I. ahmadii (a new taxon) and I. leptocystis,

were collected from moist temperate forests of Pakistan and identified based on molecular

phylogenetic analysis and assessment of macro- and micro-morphological characters.

Inocybe ahmadii is described as new based on its smooth phaseoliform to sub-amygdaliform

basidiospores, pinkish brown lamellae, white stipe surface that is brownish at the apex,

broadly lageniform hyaline pleurocystidia, and similar but more frequent cheilocystidia. Our

I. leptocystis specimen, molecularly supported as conspecific with specimens from Austria,

China, Italy, and Finland, represents a first record of the species from Pakistan.

Key worps—conifers, Inocybaceae, Inocybe sensu stricto, Inocybe subg. Inocybe, nrITS

Introduction

Inocybe (Basidiomycota, Agaricales, Inocybaceae) is a highly diverse genus

of ectomycorrhizal fungi comprising 500-750 species, primarily in temperate

areas. The number continues to increase due to new discoveries (e.g., Matheny

et al. 2009, Alvarado et al. 2010, Bougher & Matheny 2011, Bougher et al. 2012,

Kokkonen & Vauras, 2012, Matheny et al. 2012, Fan & Bau 2013, Braaten et al.

2014, Fan & Bau 2014, Latha et al. 2015, Esteve-Raventos et al. 2015, Jabeen et

al. 2016). Twenty-eight species of Inocybe have been reported from Pakistan

(Ahmad et al. 1997, Sultana et al. 2011, Saba et al. 2015, Jabeen et al. 2016).

258 ... Faroogi & al.

The species within Inocybe can be recognized by their often small-

to medium-sized basidiomata with spermatic, earthy, bitter almond,

pelargonium, or fruit-like smell (although many species do not have any

distinct smell), radially fibrous to cracked and often brownish pileus, and

smooth, spinose, nodulose, or angular yellowish brown basidiospores (Solak

et al. 2009). Matheny et al. (2009) subdivided Inocybe into seven major clades

or lineages based on DNA sequence analyses and morphology.

Taxa within Inocybe subg. Inocybe [= Inocybe s.str.] typically have angular/

nodulose or smooth spores, thick-walled pleurocystidia, a cortina, and a non-

to only slightly pruinose stipe apex. Our study characterizes and identifies two

species within Inocybe s.str. based on morphological and internal transcribed

spacer (ITS) nrDNA analyses.

Materials & methods

Sampling site

Sampling was carried out during the 2014 and 2015 monsoon periods in Khanspur

and Miandam of Khyber Pakhtunkhwa province, Pakistan.

Miandam is located in northeastern Swat district, a lush green valley surrounded

by mountain peaks ranging from 4500 to over 6000 m a.s.l. and mostly covered with

permanent snow. The region has a humid climate with average annual rainfall exceeding

1000 mm. Summers are mild: the hottest month is June, with mean 33°C maximum and

16°C minimum temperatures, and a rainfall of 120-132 mm in July-August. January,

with mean -2°C minimum and 11°C maximum temperatures, is the coldest month

(Sher et al. 2012).

Khanspur in Abbottabad district lies in the moist temperate western Himalayan

mountain range (3000-4000 m a.s.1.; Khan 1999). The mean annual temperature is 10°C

(Malik & Sukhera 2012). The area is characterized by cool summers and cold winters,

with a 3°C mean winter temperature during December and January and 26°C mean

summer temperature during June to September; mean annual rainfall is 1200 mm with

57% humidity (Ahmed et al. 2006).

The vegetation of both regions is dominated by coniferous trees (Cedrus, Picea, Pinus,

Taxus) with deciduous trees (Juglans, Populus, Quercus, Salix) also present (Champion

et al. 1965; Sheikh 1993)

Collection, isolation, and morphological study

Basidiomata were collected, tagged with their assigned sample code and collection

date, and photographed. Fresh macroscopical characters were documented in detail.

Colors were designated following the Munsell Soil Color Charts (1975). For subsequent

anatomical and molecular analyses, specimens were dried under an electric fan heater

to prevent any potential fungal attack or decomposition. Tissues were mounted in

5% KOH and spores, basidia, cystidia, pileipellis, and stipitipellis were examined

under a MX4300H light microscope (Meiji Techno Co., Ltd., Japan). Line drawings

Inocybe ahmadii sp. nov. (Pakistan) ... 259

of microscopic characters were made with a Leitz camera lucida (Wetzlar, Germany).

For each species, 20 basidiospores / from 1 specimen / from 1 collection (indicated

as: [20/1/1]) were measured. Dimensions are given as (a—)b-c(-d), with the range

b-c containing at least 90% of the measured values and extreme values (a & d) given

in parentheses. Abbreviations: avl = average spore length, avw = average width,

Q (quotient) = spore length/width ratio, and avQ = average quotient of all basidiospores

(Liang & Yang 2011). Specimens were deposited in the Herbarium, University of the

Punjab, Lahore, Pakistan (LAH).

DNA extraction, amplification & sequencing

Genomic DNA was extracted from a small piece (30-50 mg) of dried pileus

and lamellae using 2% CTAB extraction buffer following the Bruns (1995) protocol

with some modifications (300 ul of chloroform was combined with iso-amylalcohol

(24:1), centrifuge conditions altered to 13.2 rpm for 20 min at 4°C, and the DNA

pellet was resuspended in 50 ul of deionized water). The nrDNA ITS region was

amplified with ITSI1F as forward primer and ITS4 as reverse primer (White et al.

1990) using the Extract-N-Amp PCR kit (Sigma Aldrich, St. Louis, MO, U.S.A.). The

DNA was amplified in 20 ul reaction volumes following thermocycler conditions

in Gardes & Bruns (1993). Agarose gel electrophoresis was performed to visualize

amplified fragments of DNA in the Uvtec gel documentation system (Avebury House,

Cambridge, UK) using default settings (Sambrook & Russell 2001). PCR products

were directly sequenced bi-directionally using the same primer set at Macrogen Inc.,

Republic of South Korea.

Molecular phylogenetic analysis

The generated sequences were assembled using BioEdit software. Nucleotide

sequences were compared with the Basic Local Alignment Search Tool (BLAST) at

National Center for Biotechnology Information (NCBI; U.S.A. database). Closely

related sequences with identity greater than 90% and maximum query cover with 0.0

E value were retrieved from GenBank to reconstruct an ITS phylogeny. All sequences

were aligned using MUSCLE (http://www.ebi.ac.uk/tools/msa/muscle). The alignment

portion from CATTA to GACCT between the conserved motifs GAT to CAAA was

included in analysis. MEGA6 (Tamura et al. 2013) was used for phylogenetic tree

construction. A Maximum Likelihood (ML) tree of nrITS sequences was constructed

using a General Time Reversible (GTR) model and _nearest-neighbor-interchange

(NNI) as a ML heuristic search method. 1000 bootstrap replicates were performed.

Inocybe ahmadii Faroogi, Niazi & Khalid, sp. nov. Figs 1, 2

MycoBAnk MB 817163

Differs from Inocybe glabripes by its lageniform pleurocystidia and numerous lageniform

cheilocystidia.

Type: Pakistan, Khyber Pakhtunkhwa province, Abbottabad district, Khanspur,

3000-4000 m a.s.l., solitary on ground under Pinus wallichiana A.B. Jacks., 14 August

2015, Ayesha Farooqi AF14 (Holotype: LAH14215; GenBank KX254462).

260 ... Faroogi & al.

Fic. 1. Inocybe ahmadii basidiome (LAH14215, holotype). Scale bars = 1 cm.

EryMoLoGy—tThe specific epithet refers to Sultan Ahmad, the pioneer mycologist of

Pakistan, for his contributions in the field of mycology.

BASIDIOMA small, <46 mm high, pale brown when dry. PILEus small, 21 mm

diam, plano-convex to applanate, umbonate, umbo rather low and obtuse; dark

pinkish brown (5YR1/4) at the centre, lighter towards margins, light pinkish

brown (5YR4/6); surface dry, with appressed scales, weakly fibrillose towards

the margin, slightly rimulose, edges appendiculate with whitish fibrils; context

very thin, whitish. LAMELLAE adnexed, moderately crowded, alternating with

lamellulae, 0.1 mm wide, buff (7.5YR9/4), turning brown when dry, edges

entire to slightly fimbriate. Stipe 42 x 3 mm, central, even, surface pruinose

at the apex, fibrillose towards the base, brownish at the apex, white to greyish

towards the base, cortina present and fugacious. CONTEXT loosely stuffed,

interior tubular. ODourR not recorded.

BASIDIOSPORES [20/1/1] (7.8-)7.9-9.0(-9.2) x (4.3-)4.7-4.9(-5.0) um, avl x

avw = 8.4 x 4.7 um, Q = (1.5-)1.6-1.8(-2.0), avQ = 1.7, ellipsoid; phaseoliform

Inocybe ahmadii sp. nov. (Pakistan) ... 261

Fic. 2. Inocybe ahmadii (LAH14215, holotype). A. basidiospores; B. basidia;

C. cheilocystidia; D. pleurocystidia; E. caulocystidia; E hyphae from pileipellis;

G. stipitipellis Scale bars: A-E = 20 um; E.G = 40 um. Drawings by Ayesha Faroogi.

262 ... Faroogi & al.

to subamygdaliform, apiculate, mostly with obtuse apices, pale yellow brown,

uniguttulate. Bastp1a 19.5-25 x 6.5-8.5 um, avl x avw = 22.5 x 7.5 um,

cylindrical to clavate, 2-4-spored, hyaline. PLEURocystTip1a 51.5-65 x 12-

17.5 um, avl x avw = 59 x 14.5 um, narrowly to rather broadly lageniform with

obtuse apices and a small pedicel, wall <3 um thick. CHEILocystTip1a 49-55

x 13-16.5 um, avl x avw = 52 x 15 um, similar to pleurocystidia but generally

shorter and wider, moderately encrusted at the apex, rarely tapering at the

base, hyaline, paracystidia rare. CAuLocystTIbIA slender, cylindrical, rarely

subfusiform, moderately thick-walled, present only at stipe apex. PILEIPELLIS

9-24 um thick (av = 12.5 um), a trichoderm, hyphae ochraceous, clamp

connections present. STIPITIPELLIS 6-14 um thick (av = 9.0 um), composed

of cylindrical hyphae, regularly arranged, thin-walled, clamp connections

present.

Inocybe leptocystis G.F. Atk., Am. J. Bot. 5: 212 (1918) Fras 3,4

Piteus 16 mm diam, yellowish brown (5YR5/8) to brown (5YR9/12)

towards the margin, smooth to finely fibrillose, umbonate, margins incurved.

LAMELLAE moderately wide, adnate, 2-4 mm wide, light brown (2.5YR2/6),

edges fimbriate. Stipe 45 x 7 mm, cylindrical, brown (2.5Y3/6) to whitish

(2.5YR9/12), slightly enlarged at the base; surface smooth, nowhere pruinose;

central, narrower towards the apex. CONTEXT white to yellowish towards the

stipe base. ODOUR weakly spermatic.

BASIDIOSPORES [20/1/1] (9.0-)10.1-12.5(-12.7) x (5.2-)5.4-6.4(-6.6)

um, avl x avw =11.6 x 6.0 um, Q = (1.5-)1.7-2.2(-2.3), avQ = 1.9, smooth,

subamygdaliform, yellowish brown, apiculate, sometimes slightly depressed

above the apiculus, mostly uniguttulate. Bastp1A 28.3-35.4 x 7.1-9.2 um, avl x

avw = 33.2 x 8.6 um, cylindrical, hyaline to yellow, 4-spored. PLEUROCYSTIDIA

35-60 x 16-23 um, avl x avw = 53 x 18 um, with median constriction and

clavate to utriform, apices obtuse or subcapitate, thin-walled, hyaline to

light yellow. CHEILOCYSTIDIA 52-68 x 12-24 um, avl x avw = 62 x 17.5 um,

clavate, thin-walled, hyaline to light yellow, in various forms, rather dispersed.

CAULOCYSTIDIA not observed. PILEIPELLIS 6-10.5 um thick (av = 9.8 um), a

cutis, hyaline, composed of irregular hyphae, walls yellowish brown, clamp

connections present. STIPITIPELLIS 7-9 um thick (av = 8.2 um), a cutis of

cylindrical hyphae, yellowish brown pigment incrusted, septate.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA PROVINCE, Swat

District, Miandam at 2000 m a.s.l, under conifers, solitary, 23 Aug 2014, Sana Jabeen

FA-5 (LAH35112; GenBank KX254461).

Inocybe ahmadii sp. nov. (Pakistan) ... 263

Molecular phylogenetic results

Sequences of the Inocybe ahmadii PCR products using ITS1F and ITS4 were

trimmed at the conserved motifs to produce consensus sequences of 643 base

pairs. BLAST searches of Inocybe ahmadii ITS1F and ITS4 sequences revealed

93-91% identity with Inocybe sp. (JF908230) from Italy listed as “Inocybe

rufulovelata’, an unpublished name, in Osmundson et al. (2013) and I. glabripes

Ricken (AJ889952) from Denmark with 94-100% query cover and 0.0 E value.

Sequencing of the Inocybe leptocystis PCR product using the same set of

primers yielded 727-809 base pairs, which were trimmed at the conserved

motifs to produce consensus sequences of 637 base pairs. The BLAST revealed a

99% similarity with I. leptocystis from China (KU359777) and Italy (JF908158)

and a 95-94% similarity with a different species identified as I. leptocystis from

China (KJ810596) and Sweden (AM882801). It therefore appears that the label

264 ... Faroogi & al.

—

Fic. 4. Inocybe leptocystis (LAH35112). A. basidiospores; B. basidia; C. pleurocystidia;

D. stipitipellis; E. pileipellis. Scale bars = 10 um. Drawings by Sana Jabeen.

Inocybe ahmadii sp. nov. (Pakistan) ... 265

“I. leptocystis” has been applied to sequences representing at least two different

species.

We compared our ITS sequences with 44 sequences retrieved from

GenBank. Inocybe dunensis P.D. Orton (JF908262) and I. hirculus Vauras

(AM882723) were chosen as outgroup (Fic. 5). After alignment, a 638

character long matrix was obtained, of which 352 characters were conserved,

271 variable, 194 parsimony-informative, and 76 singletons. Three major

clades were formed within the ingroup sequences. Inocybe ahmadii clustered

with Inocybe spp. (HG796992, JF908193) and I. glabripes Ricken in clade I but

formed its own lineage on an isolated branch. Inocybe leptocystis clustered in

clade II with I. aeruginascens Babos (labeled as “I. bresadolae”) and I. hystrix

(Fr.) P. Karst.

Discussion

Our analysis of sequences from Inocybe s.str. (clade Inocybe; Latha

et al. 2016) indicated three clades I-III. Typical characters include

presence of thick-walled pleurocystidia, two kinds of cheilocystidia

(both pleurocystidioid cheilocystidia and clavate to sphaeropedunculate

paracystidia), smooth or angular or nodulose spores with fairly large

apiculi; caulocystidia and a cortina may be either present (over a variable

stipe length) or absent (Kuyper 1986, as I. subg. Inocybe).

Clade I, which comprised 26 sequences split into two subgroups, is

characterized by species showing a brown to pale brown or ochraceous

buff colored pileus (excepting the distinctively purple brown or lilaceous

I. lanatopurpurea Esteve-Rav. & G. Moreno), a stipe that is whitish when

young then brownish, cortinate young specimens, subamygdaliform

spores, and apically encrusted thick-walled pleurocystidia. Within clade I,

the new species I. ahmadii is well separated from the other taxa, forming

its own lineage with notably long branch. This phylogenetic separation

is supported by its morphological differences, e.g., buff brown lamellae

with pinkish tinge, a whitish stipe surface with a brownish apex, ellipsoid

and phaseoliform to subamygdaliform spores, apically encrusted and

narrowly to broadly lageniform pleurocystidia, and generally shorter and

broader cheilocystidia. Inocybe ahmadii forms a subgroup with Inocybe sp.

HG796992 (Pakistan) and JF908193 (U.S.A.) and I. glabripes AJ889952,

AM882807 (Sweden) and JF908239 (U.S.A.), but a 86% ML bootstrap

value well supports its separation from these taxa. Inocybe glabripes is

distinguished by its olivaceous tinged ochraceous brown lamellae, its

266 ... Faroogi & al.

HQ604149 J. flocculosa var. focculosa

HQ604150 I. facera var. lacera

EU525950 £. cf fanuginosa

HQ604145 J. focculosa var. focculosa

HQ604146 © /acera var. helobia

HQ604143 L Aotsoniana

EU525985 I. cf lanuginosa

HQ604148 I. flecculosa var. flocculosa

HQ604152 I. facera var. lacéra

HQ604141 £. flocculosa var. Hocculosa

HQ604137 J. nitidiuscula

FN669213 Z. sp.

2 K}432291 f. anatopurpurea

KJ432289 £ fanatopurpurea

HQ604136 I. pseudodestricta

HQ604134 £ cf hotsonana

Clade I

971 149604133 L. nitidiuscula

K3432292 I. lavandulochiora

FN550925 J. subnudipes

100] amgs2809 Z subnudipes

49 80

HG796992 J sp.

JF908193 I. sp.

@ KX254462 L ahmadii

AJ889952 I cf olabripes

AM882807 L. glabripes

794+ JF908239 £ glabripes

@ KX254461 L septocystis

87) U359777 1 keptocystis

- )F908158 £ leptocystis

AM882801 I. leptocystis

99 | KI8105961. leptocystis

98 45

GU949589 Z aff hystrix | Cadet

JF908130 L. bresadolze

AM882811 £ Aystrix

78) Gusa9sss Z pystrix

57] amgs28i2 ZL. hystrix

35) JF908194 2 phaeoleuca

JF908119 1. splendens

JX630888 £. Gallingiorum

JX630342 £ Gallingiorum

Clade IIT

JX630341 £ gallingiorum

JX630649 £ Gallingioruns

66 JX630648 J. Gallingiorum

3! JX630604 F. gallingiorum

AM882723 J. hirculus

0.05

JF908262 L dunensis

Outgroup

Jnocybe s.str.

Fig. 5. The evolutionary history of species of Inocybe sensu stricto inferred by using the Maximum

Likelihood method based on the General Time Reversible model in MEGA6. The tree with the

highest log likelihood (-3363.8870) is shown. The tree is drawn to scale, with branch lengths

measured in the number of substitutions per site. Bootstrap values are indicated above or below

branches. The analysis involved 46 nucleotide sequences. There were a total of 630 positions in the

final dataset. The two Pakistani collections have b

een labeled with @ and highlighted.

Inocybe ahmadii sp. nov. (Pakistan) ... 267

ochraceous or honey brown stipe surface, subamygdaliform spores,

cylindrical to subfusiform pleurocystidia, and infrequent cheilocystidia.

All these morphological characters separate I. ahmadii from I. glabripes,

supporting its designation as a new species.

Clade II included 10 sequences, split into two subgroups, one of which

clustered our Pakistani I. leptocystis sequence with four other I. leptocystis

sequences. Species of clade II can be distinguished morphologically from clade

I by their lack of caulocystidia and presence of thin-walled pleurocystidia.

Inocybe leptocystis has been reported from several different temperate

regions: the U.S.A. (Atkinson 1918, Kobayashi 2002), Asia (China; KU359777,

Fan & Bau, unpublished), and Europe (Austria, Kobayashi 2002; Finland,

Kobayashi 2002 and Matheny 2005; Italy, Osmundson et al. 2013; Sweden,

Ryberg 2009).

The sequence data suggest two different taxa are labeled as “I. leptocystis”

within clade II: group (1) from China (KU359777) and Italy (JF908158); and

group (2) from Sweden (AM882801) and China (KJ810596). Morphologically,

the Pakistani collection corresponds with the descriptions of the collections

from Austria and China (Kobayashi 2002), and the sequence from the

Pakistani collection clusters with the group (1) sequences from China and

Italy. This group (1) subclade is represented by species having an ochraceous

brown to buff pileus, non-pruinose smooth stipe, thin-walled, subutriform to

clavate pleurocystidia (except I. hystrix, which has cylindrical to sublageniform

pleurocystidia).

Acknowledgments

We are highly indebted to University of the Punjab, Lahore, Pakistan for providing

funds under research project grant for the fiscal year 2015-2016. Thanks to Higher

Education Commission, Pakistan for funding to fourth author under the Indigenous

PhD Fellowships for 5000 Scholars, HEC (Phase-II). The authors are grateful to Dr

P. Brandon Matheny (Department of Ecology and Evolutionary Biology, University of

Tennessee, Knoxville, U.S.A.), Dr Martin Ryberg (Department of Organismal Biology,

Uppsala University, Sweden), and Balint Dima (Department of Plant Anatomy, Institute

of Biology, Edtvés Lorand University, Hungary) for valuable comments and suggestions

to improve the manuscript.

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MY COTAXON

April-June 2017— Volume 132, pp. 271-279

https://doi.org/10.5248/132.271

Corynespora titarpaniensis sp. nov.,

on Lepidagathis from Central India

PRAKASH KUSHWAHA!', RAGHVENDRA SINGH * & SHAMBHU KUMAR?

"Department of Botany, School of Biological Sciences, Dr. Harisingh Gour University,

Sagar, M.P., India 470003

*Centre of Advanced Study in Botany, Institute of Science, Banaras Hindu University,

Varanasi, U.P, India 221005

*Department of Forest Pathology, Kerala Forest Research Institute,

Peechi, Thrissur, Kerala, India 680653

*CORRESPONDENCE TO: drsinghtaxon@gmail.com

ABSTRACT—A new asexual fungus Corynespora titarpaniensis, discovered on Lepidagathis

sp. (Acanthaceae) from Madhya Pradesh, India, is described, illustrated, and compared with

closely similar species.

Key worps—biodiversity, Corynesporascaceae, foliar diseases, phytopathogenic fungi,

taxonomy

Introduction

Corynespora was described by Giissow (1906), and almost 200 species have

been recorded (Index Fungorum 2016). Indian mycologists have described

several novel species in the past 15 years (Dubey & Rai 2003; Jain et al. 2002;

Kumar et al. 2007, 2008, 2012, 2013; Meenu & Kamal 1998; Meenu et al. 1997,

1998; Pal et al. 2007; Sharma et al. 2002a,b, 2003, 2005; Singh & Kamal 2012;

Singh et al. 2000a,b, 2007a,b, 2013, 2014; Verma et al. 2008). Synoptic tables for

the main morphological features of Corynespora spp. have been provided by

McKenzie (2010) and Siboe et al. (1999).

In 2016, during routine survey for foliicolous fungi, an interesting fungus

causing a serious disease was collected on living leaves of Lepidagathis sp. from

a village in Madhya Pradesh. Based on morphological characteristics such as

272 ... Kushwaha, Singh & Kumar

colouration and septation in conidia and conidiophores, as well as mode of

conidial ontogeny, this fungus was assigned as a new species of Corynespora.

A detailed description and illustrations of the new fungus are presented here.

Materials & methods

Fresh leaf specimens with disease symptoms were collected during the course of field

trips. The samples were placed in zip-lock polythene bags, taken to the laboratory, and

treated according to Castafieda-Ruiz (2005). Detailed observations of morphological

characters were carried out using a Leica light microscope. Specimens were prepared

for microscopical observation by hand sectioning and scraping; the sections were

mounted in lactophenol cotton-blue. Measurements were made of 30 conidia, hila, and

conidiophores. Line drawings were prepared at a magnification of 400x. Photographs of

colonies on leaves and microphotographs of fungal propagules were taken using a Sony

DSC-5730 camera. The holotype is deposited in the Ajrekar Mycological Herbarium,

Ajrekar Research Institute (ARI), Pune, India (AMH), and isotype material is deposited

in the Mycological Herbarium of the Department of Botany of Dr. Harisingh Gour

University Sagar, M.P., India (MH-DHSGU).

Corynespora titarpaniensis P. Kushwaha, Raghv. Singh & Sh. Kumar,

sp. nov. FIGs 1-4

MycoBank MB 818169

Differs from Corynespora barleriicola and C. peristrophicola by its larger, solitary conidia

with more numerous septa.

Type: India, Madhya Pradesh, Sagar, Deori, Titarpani village, 23.3898°N 79.0165°E,

on living leaves of Lepidagathis sp. (Acanthaceae), Jan. 2016, coll. Prakash Kushwaha

(Holotype, AMH 9786; isotype, MH-DHSGU 188).

ErymMo_oey: Latin, titarpaniensis, referring to the type locality.

INFECTION SPOTS amphigenous, scattered, initially circular to subcircular,

1-9 mm in diam., later becoming irregular and spreading over entire

leaf surface, more or less necrotic, brown to blackish brown. COLONIES

amphiphyllous, effuse, brown to blackish brown. Myce.ium internal,

thin-walled, branched, smooth, brown to blackish brown. STROMATA

absent or rarely present. CONIDIOPHORES macronematous, arising singly

or rarely in groups of 2-7, cylindrical, erect to procumbent, straight to

flexuous, smooth, unbranched, thick-walled, 5-16-septate, constricted

at septa, medium brown to blackish brown, 155-500 x 5-11.5 um, with

numerous successive cylindrical enteroblastic percurrent extensions, basal

cell swollen. CONIDIOGENOUS CELLS monotretic, integrated, terminal, scars

unthickened. Conrp1a solitary, smooth, simple, thin-walled, straight to

slightly curved or wavy in margin, obclavate-cylindrical, apex obtuse to

Corynespora titarpaniensis sp. nov. (India) ... 273

Fic. 1. Corynespora titarpaniensis on Lepidagathis sp. a, b. Infected host plant. c, d. Infection spots

on upper surface. e, f. Infection spots on lower surface. Scale bars = 10 mm.

rounded, 5-35-distoseptate with some transverse band-like eusepta, with

some deep constrictions, 50-340 x 5-20 um, olivaceous to light brown,

hilum rounded to obconico-truncate, thickened, 3-5 um diam.

DiscussIon—Two other Corynespora species have been described from

acanthaceous hosts: C. barleriicola N. Sharma et al. and C. peristrophicola

Raghv. Singh & Kamal. The conidia of C. barleriicola (41-246 x 10-18.5 um,

274 ... Kushwaha, Singh & Kumar

Fic. 2. Corynespora titarpaniensis (holotype, AMH 9786). a-f. Conidia. g-i. Germinating conidia.

Scale bars: a~c = 50 um; d-i = 20 um.

Corynespora titarpaniensis sp. nov. (India) ... 275

Fic. 3. Corynespora titarpaniensis (holotype, AMH 9786).a. Solitary conidiophore. b. Conidiophores

in fascicle. c-e. Bulbous basal cell of conidiophores. f-h. Developing conidia at conidiogenous loci.

Scale bars: a, b = 50 um; c, d = 20 um; e-h = 40 um.

276 ... Kushwaha, Singh & Kumar

Fic. 4. Corynespora titarpaniensis (holotype, AMH 9786).

a. Infection spots. b. Conidiophores. c. Conidia. d. Germinating conidia.

Corynespora titarpaniensis sp. nov. (India) ... 277

3-14-septate; Sharma et al. 2002b) and C. peristrophicola (60-135 x 5-16 um,

5-12-septate; Singh & Kamal 2012) are catenate and shorter and have fewer

septa than those of C. titarpaniensis.

The conidia of C. azadirachtiana N. Sharma et al. (32-303.5 x 7-21.5 um,

1-20-septate; Sharma et al. 2002b), C. colebrookiana N. Sharma et al. (45-330

x 6-22 um, 4-16-septate; Sharma et al. 2002b), C. catharanthicola Z.D. Jiang &

PK. Chi (140-310 x 5.5-11 um, 4-25-septate; Chi 1994) and C. proliferata Loer.

(30-300 x 9-12 um, 3-17-septate; Loerakker 1975) are catenate and have fewer

septa than C. titarpaniensis.

Corynespora leucaenae N. Sharma et al. (16-298 x 10-19 um, 1-28-septate;

Sharma et al. 2003), C. trematicola N. Sharma et al. (104-296 x 11-16 um,

1-12-septate; Sharma et al. 2002a, as “tremicola”) and C. longispora A.K. Sarbhoy

& Saikia (120-330 x 5.5-8 um, 11-24-septate; Saikia & Sarbhoy 1981) also have

conidia similar to those of C. titarpaniensis but differ by having fewer septa.

The conidia of C. smithii (Berk. & Broome) M.B. Ellis (70-410 x 12-19 um,

7-45-septate; Ellis 1957) and C. viticus Y.L. Guo (80-383 x 6-9 um, multiseptate;

Guo 1984) appear close to C. titarpaniensis but differ by their catenate,

cylindrical, longer, thick-walled conidia with more septa.

The new species also resembles C. rhododendri K. Zhang & X.G. Zhang,

which differs by its much longer, thinner, obclavate to long rostrate conidia

(180-400 x 7.5-11 um, 19-36-septate; Zhang et al. 2008).

Another similar species, C. micheliae Z.Q. Shang & X.G. Zhang differs from

C. titarpaniensis by its rostrate conidia (333-360 x 15-19 um, 12-28-septate)

with its upper cells gradually tapering to a narrow apex, a wider hilum (6-7 ttm),

and a darker brown lower cell; and by its shorter, wider conidiophores (190-210

x 9-19 um; Shang & Zhang 2007).

Acknowledgments

The authors thank Dr Rafael F. Castafteda-Ruiz and Dr Eric H.C. McKenzie for

reviewing the manuscript. We also express our deep gratitude to Dr Shaun Pennycook

for nomenclatural review and Dr Lorelei L. Norvell, Editor-in-Chief, Mycotaxon for

final acceptance of the manuscript for publication. Thanks are also due to the Curator,

AMH, Pune, for accepting the holotype specimen and providing an accession number.

We are much obliged to the Heads of Department of Botany, Dr. H.S. Gour University,

Sagar and CAS in Botany, Banaras Hindu University, Varanasi, as well as the Director,

Kerala Forest Research Institute, Peechi, for necessary facilities and support.

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Kumar S, Singh R, Pal VK. 2007. Three hitherto undescribed species of Corynespora from North-

Eastern Uttar Pradesh. Journal of Basic Applied Mycology 6(1-2): 39-43.

Kumar S, Singh R, Singh DP, Pal VK, Agarwal DK. 2008. Additions to new species of Corynespora

Giissow from North-Eastern Uttar Pradesh. Indian Phytopathology 61: 111-117.

Kumar S, Singh R, Saini DC, Kamal. 2012. A new species of Corynespora from terai forest of

northeastern Uttar Pradesh, India. Mycosphere 3: 410-412.

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Kumar S, Singh A, Singh R, Dubey NK. 2013. Corynespora bombacina causing foliar disease on

Bombax ceiba from Sonebhadra forest of Uttar Pradesh, India. Canadian Journal of Plant

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MY COTAXON

April-June 2017— Volume 132, pp. 281-287

https://doi.org/10.5248/132.281

Morganella austromontana sp. nov.

from the South Brazilian Plateau

CAMILA R. ALVES ?*, VAGNER G. CORTEZ” & ROSA M. B. DA SILVEIRA?

' Programa de Pés-graduacado em Botanica, Universidade Federal do Rio Grande do Sul,

Av. Bento Goncalves 9500, Agronomia CEP 91501-970, Porto Alegre, RS, Brazil

? Programa de Pés-graduacdo em Botanica, Departamento de Biodiversidade,

Rua Pioneiro 2153, Jardim Dallas CEP 85950-000, Palotina, PR, Brazil

* CORRESPONDENCE TO: cra.micologia@gmail.com

ABSTRACT—A new Morganella species was collected during field exploration of the

mycobiota of the South Brazilian Plateau. It is proposed here as Morganella austromontana,

distinguished by its ovoid basidiospores and its soil habitat. A complete description and

illustrations of its morphological features are accompanied by a taxonomic discussion.

KEY WworRDs—Agaricaceae, Agaricomycetidae, Araucaria angustifolia forest, puffballs,

taxonomy

Introduction

Morganella Zeller is diagnosed by small gasteroid basidiomata (puffballs)

that are globose, subglobose or depressed enclosed within a double peridium

comprising a granular, verrucose, velutinous or spiny exoperidium and an

endoperidium with an apical pore; basidiospores are globose or subglobose

(rarely ovoid), a eucapillitium is absent, and a paracapillitium is present

(Zeller 1948, Kreisel & Dring 1967). About 20 species have been included in

the genus. From Brazil, 12 species have been reported and/or described in

the last decade (Cortez et al. 2007; Trierveiler-Pereira & Baseia 2009; Barbosa

et al. 2011; Alfredo et al. 2012, 2014; Alfredo & Baseia 2014; Alves & Cortez

2013). These puffballs occur in diverse Brazilian biome habitats, including arid

environments (Barbosa et al. 2011) and seasonal ombrophilous forests (Alfredo

et al. 2012, Alves & Cortez 2013) where they grow on rotting wood among litter

and mosses.

282 ... Alves, Cortez & Silveira

During a survey of the gasteroid mycobiota in South Brazilian forests in

the Atlantic Forest biome, a new Morganella species was found growing in

the mixed ombrophilous forest typical of south Brazil. Here we describe this

species as Morganella austromontana.

Materials & methods

Specimens were gathered during field trips in the natural reserves of the Rio das

Furnas in the municipality of Alfredo Wagner (27°42’00’S 49°20’02”W), State of Santa

Catarina, and the National Park of Aparados da Serra in the municipality of Cambara

do Sul (29°08’02”S 49°59’40”W), State of Rio Grande do Sul. Both sites are covered by

Araucaria angustifolia forests and are located on the South Brazilian Plateau at altitudes

of 900-1200 m (Veloso et al. 1991). K6ppen (1948) classified the region's climate as Cfb:

mesothermic, wet (with regular rainfall throughout the year), and with mild summers.

The specimens were examined morphologically according to Miller & Miller (1988).

Colors were coded according to the color chart of Kornerup & Wanscher (1978).

Measurements and photographs were taken using a Motic BA310 microscope equipped

with a Moticam 2500 digital camera and Motic Image Plus 2.0 software. Scanning

electron microscopy (SEM) was performed at the Center of Electronic Microscopy of

the Universidade Federal do Rio Grande do Sul (CME/UFRGS) under a Jeol JSM-6060

SEM. All studied specimens are preserved at the Herbarium, Universidade Federal do

Rio Grande do Sul, Porto Alegre, Brazil (ICN). Five collections from Rio Grande do Sul

conserved in the ICN Herbarium representing species closest to M. austromontana were

also examined.

Taxonomy

Morganella austromontana C.R. Alves, Cortez & R.M. Silveira, sp. nov. Fics 1-6

MycoBAnk MB 820972

Differs from Morganella subincarnata by its lighter exoperidium, its ovoid basidiospores,

and its habitat on soil in araucariaceous forest.

Type—Brazil, Santa Catarina State: Alfredo Wagner, Reserva do Rio das Furnas,

7.1.2014, leg. C.R. Alves 94 (Holotype, ICN 181431).

ErymoLocy—The name, meaning “from the southern mountains,’ refers to the

montane type region on the South Brazilian Plateau.

Basip1oMEs 10 mmhigh, 8-20 mmdiam.,subglobose to pyriform. EXOPERIDIUM

spiny, formed of short spines (<1 mm high), numerous and densely grouped

on the top of the basidioma, light brown (6D5) when fresh, falling away as

plates detach from the endoperidium when mature. ENDOPERIDIUM papery,

smooth to areolate, white (1A1) when fresh, then yellow grey (2B2) after dried.

STERILE BASE poorly developed to present, reaching 3 mm high, yellowish

grey (2B2), with scattered, white and thin rhizomorphs (<1 mm thickness).

Morganella austromontana sp. nov. (Brazil) ... 283

Fics 1-3. Morganella austromontana (holotype ICN 181431). 1. Basidiome in situ. 2. Dried

basidiomata. 3. Paracapillitium and basidiospores. Scale bars: 1, 2 = 10 mm; 3 = 10 um.

GLEBA cottony, brownish grey (2D2). SUBGLEBA present, <3 mm high, cellular,

greyish yellow (1B3). Bastp1osporgss 4-5 x 3-4 um, ovoid to subglobose, pale

green in 5% KOH, surface verrucose under light microscope, spiny under SEM;

284 ... Alves, Cortez & Silveira

spines cone-shaped, <2 um high. PARACAPILLITIUM 7-10 um diam., inflated,

with numerous small pores c. 1 um diam., amorphous matter surrounding

the hyphae, septa regular, sometimes, with dichotomous tips. ExOPERIDIUM

hyphae 10-26 x 8-15 um diam., globose, subglobose or pyriform, hyaline to

pale yellow, walls thin, about 1 um. MycoscLEREIDs 25-80 x 7-13 um diam.,

irregular in shape, hyaline, walls thick (3 um) and smooth.

ECOLOGY & DISTRIBUTION—Basidiomata solitary to gregarious, growing

on soil, among a few herbs on the trail board in the Mixed Ombrophilous

Forest, with Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae; Parana

pine, Brazilian pine), about 480-900 m altitude, during rainy summer. Only

known from the South Brazilian Plateau (States of Rio Grande do Sul and Santa

Catarina).

ADDITIONAL SPECIMENS EXAMINED — BRAZIL. R10 GRANDE DO SUL STATE: Cambara

do Sul, Parque Nacional dos Aparados da Serra, 13.III.2014, leg. A. Gerlach 1486

(ICN 181432); Sao Francisco de Paula, Veraneio Hampel, 8.1.2006, leg. V.G. Cortez

013/06 (ICN 154553); FLONA, 21.1V.2007, leg. V.G. Cortez 048/07 (ICN 154554);

Itaara, Parque Pinhal, 4.V.2007, leg. V.G. Cortez 075/07 (ICN 154555); Santa Maria,

FEPAGRO, 15.V.2007, leg. V.G. Cortez s.n (ICN 154556); Campus UFSM, 4.V1I.2007,

leg. G. Coelho 485-7 (ICN 154557).

ComMMENTS—Morganella austromontana is described as a new based on its

combination of mainly ovoid ornamented basidiospores (verrucose under

LM; shortly spiny under SEM) and an exoperidium densely covered with short

white spines in younger basidiomata that become pale brownish and fall as

plates in maturity. Another diagnostic feature is the relatively wide (<10 um)

diameter of paracapillitium hyphae, which is rarely greater than 5-6 um in

other species. The thick-walled mycosclereids found in the exoperidium of

M. austromontana have also been observed in M. sulcatostoma C.R. Alves &

Cortez (Alves & Cortez 2013).

Morganella austromontana can be nested among members of Morganella

sect. Subincarnata sensu Ponce de Leon (1971), a section that accommodates

species with a deciduous exoperidium and reticulate-areolate endoperidium.

Within M. sect. Subincarnata, M. costaricensis M.I. Morales, which shares

with M. austromontana an exoperidium with deciduous spines and areolate

endoperidium, can be separated by its smooth globose to subglobose

basidiospores (Morales et al. 1974, Suarez & Wright 1996).

The type species of the section, Morganella subincarnata (Peck) Kreisel &

Dring, appears related to M. austromontana based on its similar pattern of exo-

and endoperidium but differs in its darker (olivaceous brown) exoperidium,

its globose basidiospores (3.5 x 6.5 um diam.), and its preferred substrate on

Morganella austromontana sp. nov. (Brazil) ... 285

Fics 4-6. Morganella austromontana (holotype ICN 181431). 4. Basidiospores and paracapillitium.

5, 6. Basidiospores (SEM). Scale bars 4 = 10 um; 5, 6 = 1 um.

286 ... Alves, Cortez & Silveira

angiospermous wood (Bowerman 1961, as Lycoperdon subincarnatum; Coker

& Couch 1928, as Lycoperdon subincarnatum; Kreisel & Dring 1967; Ponce de

Leon 1971).

Another noteworthy feature of Morganella austromontana is the fact its

basidiomata were collected from soil, instead of rotting wood, the usual

substrate for most Morganella species. However, a soil substrate is not exclusive

to M. austromontana; for example, M. arenicola Alfredo & Baseia grows on

sandy soil (Alfredo et al. 2014).

Further fieldwork will be necessary to determine if the proposed species

is, in fact, restricted to the southern Brazilian highlands and, in particular, to

A. angustifolia forests. However, based on the current available data and

ongoing survey of the gasteroid fungi of Southern Brazil, M. austromontana

appears strongly associated with these environments.

Acknowledgments

We thank the financial support of Conselho Nacional de Desenvolvimento Cientifico

e Tecnoldgico (CNPq) and Coordenacgao de Aperfeicoamento de Pessoal de Nivel

Superior (CAPES), the reviewers Dr. Maria Luciana H. Caffot and Dr. Iuri G. Baseia for

the considerations, Instituto Chico Mendes (ICMBio) for allowing fieldwork, and Alice

C. L. Gerlach for specimen collection.

Literature cited

Alfredo DS, Baseia IG. 2014. Morganella nuda, a new puftball (Agaricaceae, Basidiomycota)

in the upland forests of the Brazilian semi-arid region. Nova Hedwigia 98: 459-466.

http://dx.doi.org/10.1127/0029-5035/2014/0164

Alfredo DS, Leite AG, Braga-Neto R, Baseia IG. 2012. Two new Morganella species from the Brazilian

Amazon rainforest. Mycosphere 3: 66-71. http://dx.doi.org/10.5943/mycosphere/3/1/8

Alfredo DS, Accioly T, Baseia IG. 2014. Morganella arenicola, a new species record from North and

Northeast Brazil. Turk. J. Bot. 38: 595-599. http://dx.doi.org/10.3906/bot-1307-68

Alves CR, Cortez VG. 2013. Morganella sulcatostoma sp. nov. (Agaricales, Basidiomycota)

from Parana State, Brazil. Nova Hedwigia 96: 409-417.

http://dx.doi.org/10.1127/0029-5035/2013/0078

Barbosa MMB, Silva MA, Cruz RHSE, Calonge FD, Baseia IG. 2011. First report of Morganella

compacta (Agaricales, Lycoperdaceae) from South America. Mycotaxon 116: 381-386.

http://dx.doi.org/10.5248/116.381

Bowerman CA. 1961. Lycoperdon in eastern Canada with special reference to the Ottawa district.

Can J Bot 39: 353-383. http://dx.doi.org/10.1139/b61-029

Coker WC, Couch JN. 1928. The gasteromycetes of the eastern United States and Canada.

University of North Carolina Press, Chapel Hill.

Cortez VG, Calonge FD, Baseia IG. 2007. Rick’s species revision 2: Lycoperdon benjaminii

recombined in Morganella. Mycotaxon 102: 425-429.

Képpen WP. 1948. Climatologia, con un estudio de los climas de la Tierra. Fondo de Cultura

Economica, México.

Morganella austromontana sp. nov. (Brazil) ... 287

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3rd ed. Eyre Methuen, London.

Kreisel H, Dring DM. 1967. An emendation of the genus Morganella Zeller (Lycoperdaceae). Feddes

Repert. 74: 109-122. http://dx.doi.org/10.1002/fedr.19670740105

Miller OK, Miller HH. 1988. Gasteromycetes: morphological and development features with keys

to the orders, families and genera. Mad River, Eureka.

Morales IM, Nassar M, Saénz JA. 1974. Lycoperdaceae of Costa Rica I. The genus Morganella. Rev.

Biol. Trop. 21: 221-227.

Ponce de Leon P. 1971. Revision of the genus Morganella (Lycoperdaceae). Fieldiana, Bot. 34: 27-44.

http://dx.doi.org/10.5962/bhI.title.2567

Suarez VL, Wright JE. 1996. South American gasteromycetes V: the genus Morganella. Mycologia

88: 655-661. http://dx.doi.org/10.2307/3761163

Trierveiler- Pereira L, Baseia IG. 2009. A checklist of the Brazilian gasteroid fungi (Basidiomycota).

http://www.mycotaxon.com/resources/checklists/trierveiler-v108-checklist.pdf

[Summary: Mycotaxon 108: 441-444. http://dx.doi.org/10.5248/108.441].

Veloso HP, Rangel-Filho ALR, Lima JCA. 1991. Classificagao da vegetacao brasileira adaptada a um

sistema universal. IBGE, Rio de Janeiro.

Zeller SM. 1948. Notes on certain gasteromycetes including two new orders. Mycologia 40:

639-668. http://dx.doi.org/10.2307/3755316

MY COTAXON

April-June 2017— Volume 132, pp. 289-297

https://doi.org/10.5248/132.289

Microidium phyllanthi-reticulati sp. nov.

on Phyllanthus reticulatus

JAMJAN MEEBOON™ & SUSUMU TAKAMATSU’

‘Graduate School of Bioresources, Mie University,

1577 Kurima-Machiya, Tsu 514-8507, Japan

* CORRESPONDENCE TO: jamjanm@yahoo.com

ABSTRACT—A new powdery mildew species of the genus Microidium (Erysiphaceae) found

on Phyllanthus reticulatus (Phyllanthaceae) in Thailand is described. Microidium phyllanthi-

reticulati [= Oidium phyllanthi var. reticulati] is characterized by having amphigenous

colonies on the host surface, long conidiophore foot-cells twisted at the base, and barrel-

shaped to cylindrical conidia with Microidium-type germ tubes. The new species differs from

the allied and morphologically very similar M. phyllanthi in producing smaller conidia and

conidiophores with longer foot-cells. Analyses of rDNA ITS + 28S sequence data support the

separation of M. phyllanthi-reticulati from M. phyllanthi. The new species is described and

illustrated in detail, and compared with other species assigned to Microidium.

Key worps—Asia, Erysiphales, phylogeny, Euphorbiaceae

Introduction

The name Oidium phyllanthi J.M. Yen was originally proposed for a powdery

mildew found on Phyllanthus urinaria in Taiwan; two types of conidia and

conidiophores were described—primary conidiophores with cylindrical to

oval conidia and secondary conidiophores producing with cylindrical conidia

(Yen 1967). Subsequently O. phyllanthi has also been recorded on Phyllanthus

acidus, P. amarus, P. niruri, P. reticulatus, and P. rheedei (Braun & Cook 2012,

as Microidium phyllanthi). Narayanswami & Ramakrishnan (1969) published

an illegitimate later homonym, O. phyllanthi Narayanas. & K. Ramakr., based

on an Indian collection on Phyllanthus niruri var. amarus [= P. amarus], but

provided only a meagre description confined to the size of conidia (14.5-20

290 ... Meeboon & Takamatsu

x 2.2-9.1 um); Hosagoudar (1991) introduced O. ramakrishnanii Hosag. as a

legitimate replacement name for this species.

Oidium subg. Microidium To-anun & S. Takam. was introduced by To-anun

et al. (2005) based on molecular phylogenetic analyses of the rDNA of 18S

and 28S regions and a morphological re-assessment of specimens of Oidium

phyllanthi found on Phyllanthus acidus, P. amarus, and P. reticulatus. The

specimens concerned were unique due to the formation of a different type of

conidial germination, designated as Microidium-type (To-anun et al. 2002).

Subsequent molecular analysis using a 18S and 28S rDNA (D1 and D2 regions)

combined dataset indicated that the sequences from O. phyllanthi on P. acidus, P.

amarus, and P. reticulatus represented a unique lineage within the Erysiphaceae

and that the sequence of a P reticulatus isolate differed from the P acidus and

P. amarus isolates by 1.38% in the 28S rDNA D1/D2 sequence (To-anun et al.

2005).

Braun & Cook (2012: 624) later raised Oidium subg. Microidium to

generic rank as Microidium (To-anun & S. Takam.) To-anun & S. Takam.

Here we present ITS + 28S analyses of Microidium on Phyllanthus, including

new sequences from 15 specimens and additional sequences retrieved from

GenBank, together with detailed morphological examinations of all of these

specimens.

Materials & methods

Morphological examination

Specimens were collected in northern Thailand between November 2004 and

January 2016. The host name, collection data, localities, and the name of the collector

were noted. Morphological examinations were carried out as outlined in Meeboon &

Takamatsu (2015a). Hyphae, conidiophores, and conidia were stripped from the leaf

surfaces with a clear adhesive tape, mounted on a microscope slide with the fungal

mycelium upper most, and examined in water using a light microscopy with phase

contrast at 20x, 40x, and 100x (oil immersion). Thirty conidia were measured from

each specimen examined.

Conidial germ tubes were examined using the method of Hirata (1942). The inner

surface cell layer of onion scales was cut with a razor in a size of 1 cm’ and stripped off

by a clean forceps. The cell layer was kept in 80% ethanol more than two weeks and

rinsed with tap water for 30 min before use. The cell layer was put on a microscope slide,

followed by removing excess water with filter paper, and powdered with the conidia.

The conidia-covered cell layer, floating on distilled water in a Petri dish, was incubated

at 20-25°C for 24h until microscopic examination.

Specimens were deposited in the herbarium of the National Museum of Nature and Science,

Tsukuba, Japan (TNS), and Mie University Mycological Herbarium, Tsu, Japan (MUMH).

Microidium phyllanthi-reticulati sp. nov. (Thailand) ... 291

Phylogenetic analysis

Whole-cell DNA was extracted by the Chelex method (Walsh et al. 1991, Hirata

& Takamatsu 1996). The nucleotide sequences of the 5’-end of the 28S rRNA gene

(including domains D1 and D2) were determined in accordance with Meeboon &

Takamatsu (2015b). The rDNA internal transcribed spacer (ITS) region including

5.88 rDNA was amplified using primers ITS5 (White et al. 1990) and p3 (Kusaba &

Tsuge 1995) for the first amplification. One microlitre (ul) of the first reaction mixture

was used for the second amplification with the partial nested primer sets ITS5 and

ITS4 (White et al. 1990). The ITS5/ITS4 fragment was subjected to sequencing using

ITS1, ITS4, T3, and T4 (Hirata & Takamatsu 1996). Representative sequences gained

in this study were deposited in DNA databases (DDBJ, EMBL, GenBank). These

sequences were aligned with the 28S rDNA D1/D2 sequence in To-anun et al. 2005

using MUSCLE (Edgar 2004) implemented in MEGA version 6 (Tamura et al. 2013).

Alignments were further manually refined using MEGA and deposited in TreeBASE

(http://www.treebase.org/) under the accession number S12714. Phylogenetic trees

were obtained from the datasets by using the maximum parsimony (MP) method

implemented in PAUP* 4.0610 (Swofford 2002) with heuristic search option using ‘tree

bisection-reconstruction (TBR) algorithm with 100 random sequence additions to find

global optimum tree. All sites were treated as unordered and unweighted, with gaps

treated as missing data. The strength of internal branches of the resulting trees was tested

with bootstrap (BS) analyses (Felsenstein 1985) using 1000 replications with step-wise

addition option set as simple. BS values 70% or higher were given. Tree scores, including

tree length, consistency index (CI), retention index (RI), and rescaled consistency index

(RC) were also calculated.

Taxonomy

Microidium phyllanthi-reticulati Meeboon & S. Takam., sp. nov. FIG. 1

MycoBank MB 800332

= Oidium phyllanthi var. reticulati N. Ahmad, A.K. Sarbhoy, Kamal & D.K.

Agarwal, Indian Phytopathol. 57: 479. 2004 [as “reticulatus”].

Differs from Microidium phyllanthi by its smaller conidia and its longer foot-cells.

TypeE—Thailand, Chiang Mai province, Chiang Mai University, on living leaves of

Phyllanthus reticulatus Poir. (Phyllanthaceae), 22 December 2004, J. Meeboon (Holotype,

TNS-F-46801; isotype, MUMH 3364; GenBank AB719952, LC259489).

EryMoLoGy—Named after the host species, Phyllanthus reticulatus.

CoLonigs amphigenous, mycelium white, effuse, confluent, forming irregular

white patches or covering the whole leaf surface (also on young stems); hyphae

hyaline, branched, septate, 3-5 um diam., thin-walled, smooth. APPRESSORIA

well developed, lobed to multilobed, opposite in pairs or single. CONIDIOPHORES

formed singly on hyphal cells, arising + centrally from the upper cell surface,

82-183 x 4-8 um, foot-cells long, cylindrical, twisted at the base, 40-118 x

292 ... Meeboon & Takamatsu

4-8 um, with a basal septum near the junction with the supporting hypha

or elevated up to 5 um, conidiophores producing 3-6 conidia in chains

(catenescent). Conrp1A cylindrical to doliiform cylindrical, (7—)15-20(-23)

x (7-)8-10(-13) um, with a large length/width ratio of 1.8-2.5, containing

oil-drop-like inclusions but devoid of conspicuous fibrosin bodies, producing

perihilar germ tubes (“on shoulders”), broadly club-shaped, terminating in

nipple-shaped or lobed appressoria (Microidium-type).

ADDITIONAL SPECIMENS EXAMINED—THAILAND, CHIANG Mat, 27 December 2002,

J. Meeboon (MUMH 3363; GenBank AB719953); 5 January 2007, J. Meeboon (MUMH

3824; GenBank AB719954). CHIANG Rat, 12 January 2008, J. Meeboon (MUMH 3823;

GenBank AB719955); NAN, 3 February 2002, S. Kom-un (MUMH 1761; GenBank

LC259486, AB120758).

COMPARATIVE SPECIMENS EXAMINED—Microidium phyllanthi: THAILAND, CHIANG

Ral, 8 March 2004, J. Meeboon (MUMH 3361; GenBank AB719943); 8 March 2004,

J. Meeboon (MUMH 3360; GenBank AB719944); 2 November 2013, J. Meeboon

(MUMH 5743; GenBank LC259492); 18 December 2015, J. Meeboon (MUMH 6624);

26 December 2016, J. Meeboon (MUMH 6880, GenBank LC259488); 3 January 2016,

J. Meeboon (MUMH 6668; GenBank LC169073, LC259490); 15 December 2014,

J. Meeboon (MUMH 6676; GenBank LC169074, LC259491); 3 November 2013,

J. Meeboon (MUMH 5744; GenBank LC259493). MAE HonG Son, 26 November 2003,

J. Meeboon (MUMH 3366; GenBank AB719945). CHIANG Mat, 13 February 2002,

J. Meeboon (MUMH 1492; GenBank AB719946); 13 February 2002, J. Meeboon (MUMH

3362; GenBank AB719947); 25 February 2004, J. Meeboon (MUMH 3357; GenBank

AB719948); 25 February 2004, J. Meeboon (MUMH 3822; GenBank AB719949);

3 February 2007, J. Meeboon (MUMH 3352; GenBank AB719950); 3 February 2007,

J. Meeboon (MUMH 1782; GenBank LC259487, AB120755). NAN, 3 February 2002,

S. Kom-un (MUMH 1778; GenBank AB719951, AB120754).

Phylogenetic analysis

The combination of ITS + 28S sequences from 20 specimens of Microidium

from Phyllanthus (including the three specimens included in the analyses of

To-anun et al. 2005: MUMH 1761, 1778, and 1782) were aligned and analysed

with Neoerysiphe galeopsidis (DC.) U. Braun (MUMH 630) as outgroup.

The alignments were carefully edited manually, with ambiguously aligned

sites deleted from further analysis. Of the remaining 1239 total characters,

1036 were constant, 108 were variable but parsimony uninformative, and 95

were parsimony-informative. The MP analysis produced about 200K equally

parsimonious trees with 211 steps. Topologies were almost consistent among

the trees except for branching orders of the terminal branches and branch

length; a typical tree is shown in Fic. 2. The sequences obtained from the 15

specimens of M. phyllanthi from P acidus and P amarus were identical. The

sequences from the five specimens from P. reticulatus formed a separate clade

Microidium phyllanthi-reticulati sp. nov. (Thailand) ... 293

Fic. 1. Microidium phyllanthi-reticulati on Phyllanthus reticulatus (holotype, TNS-F-46801):

A. Conidia with germ tubes; B. Conidia; C. Conidiophores; D. Appressoria. Scale bar = 10 um.

294 ... Meeboon & Takamatsu

with 100% bootstrap support; they differed from the sequences from the

P. acidus and P. amarus specimens by 13.2% (ITS) and 4.42% (28S).

Discussion

Braun & Cook (2012) recognized three species currently assigned to

Microidium, M. phyllanthi (J.M. Yen) To-anun & S. Takam., M. bauhiniicola

(U. Braun & Dianese) U. Braun & Dianese, and M. agatidis (E.E. Foéx) U. Braun.

Microidium phyllanthi is distinguished from M. agatidis and M. bauhiniicola by

conidiophores with foot-cells that are twisted at the base (Braun & Cook 2012).

The foot-cells of M. agatidis and M. bauhiniicola are straight, and both species

are separated based on the sizes of their foot-cells, hosts, and differences in

the geographical distribution: M. agatidis, with 15-35 um long foot-cells, was

found on Sesbania grandiflora in Asia, whereas M. bauhiniicola, with 30-90 um

long foot-cells, was found on Bauhinia spp. in South America (Braun & Cook

2012).

Ahmad et al. (2004) described a new variety, O. phyllanthi var. reticulati,

based on the size of the conidia—slightly smaller than in O. phyllanthi J.M.

Yen [= Microidium phyllanthi] and slightly larger than in the illegitimate

O. phyllanthi Narayanas. & K. Ramakr. [= Oidium ramakrishnanii]. Ahmad

et al. (2004) also emphasized that these three entities should be considered

different varieties of O. phyllanthi. The relation between Microidium phyllanthi

s. str., described from Taiwan on P urinaria, and collections on P. acidus and

P. amarus (available name: Oidium ramakrishnanii) currently cannot be

clarified due to lack of sequence data from Taiwanese or other Microidium

collections on P. urinaria.

The powdery mildew specimens found on P reticulatus in Thailand are

morphologically very similar to the original description of O. phyllanthi var.

reticulati [conidia (7—)15-20(-23) x (7-)8-10(-13) um vs. 12.7—26.7 x 6-8

um in O. phyllanthi var. reticulati] found on the same host species. The colonies

in the specimens found in Thailand grow on both sides of the leaf surface

(amphigenous), whereas in O. phyllanthi var. reticulati they were described as

often formed epiphyllously. Ahmad et al. (2004) did not describe the conidial

germination.

Compared to the Microidium species keyed out in Braun & Cook (2012),

the new species is morphologically close to M. phyllanthi based on having

conidiophore foot-cells twisted at the base. However, M. phyllanthi-reticulati

differs from M. phyllanthi, which has larger conidia (20-30(-40) x 7-11.5 um)

and shorter foot-cells 20-105 um (Braun & Cook 2012). Microidium phyllanthi

Microidium phyllanthi-reticulati sp. nov. (Thailand) ... 295

MUMH6668 ex P. amarus

MUMH6676 ex P. acidus

ITS+288 rRNA gene

2] specimens

1239 characters MUMH3361 ex P. acidus

TL=211 MUMH3360 ex P. acidus

CI = 1,0000

RI= 1.0000 MUMH3366 ex P. amarus

RC = 1.0000 MUMH1492 ex P. acidus

MUMH3357 ex P. amarus

MUMH3822 ex P. amarus

MUMH3352 ex P. amarus

MUMH6880 ex P. acidus

mw MUMHI1782 ex P. amarus

mw MUMHI1778 ex P. acidus

MUMHS5743 ex P. acidus

MUMH5744 ex P. amarus

MUMH3362 ex P. acidus

MUMH3364 [T] ex P. reticulatus

= MUMH1761 ex P. reticulatus

MUMH3363 ex P. reticulatus

MUMH3824 ex P. reticulatus

MUMH3823 ex P. reticulatus

MUMH630 N. galeopsidis

100

M. phyllanthi

M. phyHanthi-reticulati

— 5 changes

Fic. 2. Phylogenetic analysis of the nucleotide sequences of ITS + 28S regions for 21 specimens of

Microidium from Phyllanthus; the three specimens included in the analyses of To-anun et al. (2005)

are marked with ‘#’ The isotype specimen of M. phyllanthi-reticulati is indicated with ‘[T]. The tree

is one of 200K equally parsimonious trees with 211 steps, which was obtained by the parsimony

method. Gaps were treated as missing data. Horizontal branch lengths are proportional to the

number of nucleotide substitutions that were inferred to have occurred along a particular branch

of the tree. Percentage bootstrap support 250% (1000 replications) is shown on branches.

is widespread in Africa (Ghana, Mauritius) and Asia (India, Indonesia, Sri

Lanka, Taiwan, Thailand), whereas M. phyllanthi-reticulati is known only from

India and Thailand. Furthermore, the phylogenetic analyses of the ITS + 28S

regions of the Phyllanthus powdery mildews examined support segregation of

M. phyllanthi-reticulati on P. reticulatus from M. phyllanthi on P. acidus and

P. amarus (Fa. 2).

We refrain from using the name O. phyllanthi var. reticulati as basionym for

a new combination at species level since neither the varietal type material nor

any other Indian specimens were available for morphological and molecular

examinations. Additionally, the protologue cited an incorrect orthographic

296 ... Meeboon & Takamatsu

variant of the varietal epithet (“reticulatus”), and other incorrect variants

(“reticulatum” and “reticulatae”) have been listed in databases and in the

literature; the correct epithet orthography is “reticulati” (Latin genitive case

of the host epithet “reticulatus”). Nomenclaturally, our introduction of a new

species name is legitimate, since varietal names do not have priority at species

level (McNeill et al. 2012; Art. 11.2).

Acknowledgments

This work was financially supported in part by a Grant-in-Aid for Scientific Research

(No. 16K07613) from the Japan Society for the Promotion of Science, a grant from the

Institute for Fermentation, Osaka, Japan to ST; and The JSPS postdoctoral fellowship

awarded to JM. We thank Prof. Dr. Uwe Braun and Prof. Dr. Hyeon-Dong Shin for

kindly reviewing our manuscript.

Literature cited

Ahmad N, Sarbhoy, AK, Kamal, Agarwal, DK. 2004. New anamorphic powdery mildews from

India. Indian Phytopathol. 57: 478-481.

Amano (Hirata) K. 1986. Host range and geographical distribution of the powdery mildew fungi.

Japan Scientific Societies Press, Tokyo. 741 p.

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildew). CBS

Biodiversity Series 11.707 p.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution

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MY COTAXON

April-June 2017— Volume 132, pp. 299-304

https://doi.org/10.5248/132.299

Xylaria nelumboniformis sp. nov. from tropical China

Hatr-x1A MA’, LARISSA VASILYEVAT? & Yu L13*

' Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical

Agricultural Sciences, Haikou 571101, China

? Institute of Biology & Soil Science, Far East Branch of the Russian Academy of Sciences,

Vladivostok 690022, Russia

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

* CORRESPONDENCE TO: yuli966@126.com

ABSTRACT—A new pyrenomycetous fungus from the tropical forest of Hainan Province,

southern China, Xylaria nelumboniformis, is described and illustrated. The new species is

characterized by having lotus-leaf shaped fertile stromatal apices, a flattened surface with

conspicuously tomentose edges, and navicular to crescent-shaped ascospores (23-27.5 x

7.5-9 um) with a short oblique germ slit.

KEY worDs—Ascomycota, taxonomy, Xylariaceae

Introduction

Wood-rotting fungi are an important component of forest ecosystems,

where they decompose forest litter and maintain energy flow and material

circulation (Lonsdale et al. 2008). Most Xylaria species produce stromata on

decayed wood (Dennis 1956, Martin 1970, Rogers et al. 1987, Lzessoe 1987,

Van der Gucht 1995, Ju & Rogers 1999, San Martin et al. 2001, Ju et al. 2009,

Hsieh et al. 2010, Fournier et al. 2011, Fournier 2014), and they are critical for

maintaining species diversity of these ecosystems (Rogers 1979, 2000; Whalley

1996; Osono et al. 2011). New Xylaria species and records have been described

after recent investigations on wood-rotting fungi in subtropical and tropical

forests in China (Ma et al. 2011, 2012, 2013a,b,c). This paper reports a new

species of Xylaria based on two specimens on the dead tree stump during a

survey of xylariaceous fungi in southern China. They differ morphologically

300 ... Ma, Vasilyevat & Li

from existing Xylaria species, so we describe and illustrate them as a new

species.

Materials & methods

Voucher specimens are deposited in the Fungarium of Institute of Tropical Bioscience

and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, China

(FCATAS). Microscopical features and measurements were determined from tissues

mounted in water and Melzer’s iodine reagent. The photographs of asci, ascal apical ring,

and ascospores were taken using a Keyence VHX-600E microscope, and photographs

of the stromatal surface were taken with a $70 Canon camera attached to a ZSA30w

microscope.

Taxonomy

Xylaria nelumboniformis Hai X. Ma, Lar.N. Vassiljeva & Yu Li, sp.nov. Fis 1, 2

MycoBank MB 817858

Differs from Xylaria psamathos by its tomentose stromata, larger ascospores, and short

oblique germ slit.

Type: China, Hainan Province: Diaoluoshan Natural Reserve, 18°46’N 109°50’E, alt. 500

m, on dead stump, 10 Nov. 2012, Haixia Ma, Col. 16 (Holotype, FCATAS 547).

Erymo.ocy: The epithet refers to the “lotus leaf” shape of the fertile part of the stromata.

STROMATA unbranched, upright, with applanate surface of fertile apices,

4-22 mm tall overall, short or long stipitate; fertile parts 4-10 mm diam. x

1.5-3 mm thick, nearly lotus leaf-shaped to irregularly discoid, externally

greyish-black to black, with greyish-white sloughing scales and tomentose,

especiallyontheedges;surfaceplaneorwithinconspicuousperithecialmounds,

finely cracked and tomentose, internally white, woody, texture hard. Stipes

1.5-18 x 1-1.5mm, terete, inconspicuously tomentose, sometimes contorted,

arising from a slightly enlarged base. Perithecia subglobose, 600-900 um

diam. Ostioles slightly prominent. Asci eight-spored, uniseriate, cylindrical,

long-stipitate, (180-—)190-—240(-—260) um long overall, the spore-bearing parts

150-170 x 9-10 um, the stipes 30-110 um long, with apical apparatus bluing

in Melzer’s reagent, urn-shaped to nearly rectangular, (8-)10-12(-15) x

4—5 um. Ascospores brown to dark brown, unicellular, ellipsoid-inequilateral

to navicular to crescent-shaped, with narrowly to broadly rounded ends,

smooth, (21.5-)23-27.5(-31.5) x (7-)7.5-9(-10) um, with short oblique

germ slit.

ADDITIONAL SPECIMEN EXAMINED: CHINA, HAINAN PROVINCE: Diaoluoshan Natural

Reserve, 18°46’N 109°50’E, alt. 500 m, on dead stump, 12 Jul. 2013, Haixia Ma (FCATAS

612).

Xylaria nelumboniformis sp. nov. (China) ... 301

Fic. 1. Xylaria nelumboniformis (holotype, FCATAS 547). a—-c. Stromata; d, f. Stromatal surface;

e. Mature stroma in vertical section. Scale bars: a, c = 6 mm; b = 2 mm; d, e= 1 mm; f= 0.2 mm.

ComMENts —Xylaria nelumboniformis is easily separated from other Xylaria

species by its lotus-leaf shaped or irregularly discoid flattened stromata that

are conspicuously tomentose on the edges. This species is strikingly similar

in stromatal morphology to X. psamathos Boise, which differs by its smooth

302 ... Ma, Vasilyevat & Li

Fic. 2. Xylaria nelumboniformis (holotype, FCATAS 547). a. Asci and paraphyses; b. Ascus apical

ring; c. Ascospores; d. Germ slit. Scale bars: a = 40 um; b, c = 10 um; d = 8 um.

stromata, its smaller ascospores (11-13 x 4-6 um) with a straight germ-slit

extending the full length of the spore, its flattened-cylindrical ascus rings, and

its habitat (dead grass culms in a North Carolina saltmarsh; Boise 1982).

Xylaria nelumboniformis is somewhat similar to X. crozonensis P. Leroy &

Mornand and X. michoacana F. San Martin et al. in stromatal morphology,

but X. crozonensis has cracked smooth stromata that are narrowly connected

to the substrate and smaller ascospores (13.3-15.6 x 7.6-8.8 um) with a

straight inconspicuous germ slit (Leroy & Mornand 2004, Fournier 2014).

Xylaria michoacana is distinguished by cracked smooth stromata that are

narrowly connected to the substrate and slightly smaller subglobose ascospores

(21.5-24.5 x 14-16 um; San Martin et al. 2001).

Acknowledgments

We greatly appreciate Dr. A.J. Whalley of Liverpool John Moores University and

Dr. Nian-Kai Zeng of Hainan Medical University for reviewing the presubmission

Xylaria nelumboniformis sp. nov. (China) ... 303

manuscript and giving helpful comments and suggestions. This study was supported by

the National Natural Science Foundation of China (no. 31400012) and National Basic

Research Priorities Program of China (no. 2013FY110400).

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Europe. Mycological Progress 10: 33-52. http://dx.doi.org/10.1007/s11557-010-0671-8

Hsieh HM, Lin CR, Fang MJ, Rogers JD, Fournier J, Lechat C, Ju YM. 2010. Phylogenetic status

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957-969. http://dx.doi.org/10.1016/j.ympev.2009.12.015

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343-440.

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in the China Mainland. Austrian Journal of Mycology 21: 61-67.

Ma HX, Vasilyeva LN, Li Y. 2013a. The genus Xylaria in the south of China - 3. X. atroglbosa sp.

nov. Mycotaxon 119 :381-384. http://dx.doi.org/10.5248/119.381

Ma HX, Vasilyeva LN, Li Y. 2013b. The genus Xylaria in the south of China - 4. X. hemisphaerica

sp. nov. from Yunnan Province. Mycosystema 32(4): 602-605.

Ma HX, Vasilyeva LN, Li Y. 2013c. The genus Xylaria (Xylariaceae) in the south of China - 6. Anew

Xylaria species based on morphological and molecular characters. Phytotaxa 147(2): 48-54.

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Osono T, To-Anun C, Hagiwara Y & Hirose D. 2011. Decomposition of wood, petiole and leaf litter

by Xylaria species. Fungal Ecology 4: 210-218. http://dx.doi.org/10.1016/j.funeco.2010.11.003

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1-42. http://dx.doi.org/10.2307/3759218

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Rogers JD, Callan BE & Samuels GJ. 1987. The Xylaria of the rain forests of North Sulawesi

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associated with oaks in Mexico. Mycotaxon 79: 337-360.

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http://dx.doi.org/10.1016/s0953-7562(96)80042-6

MY COTAXON

April-June 2017— Volume 132, pp. 305-315

https://doi.org/10.5248/132.305

Neozygites linanensis sp. nov., a fungal pathogen

infecting bamboo aphids in southeast China

XIANG ZHOU ™, CRISTIAN MONTALVA ’,

NOLBERTO ARISMENDI * & FEI HONG *

' National Joint Local Engineering Laboratory of Biopesticide High-Efficient Preparation,

School of Forestry and Biotechnology, Zhejiang Agricultural and Forestry University,

Linan 311300, Peoples Republic of China

’ Facultad de Ciencias, Instituto de Bioquimica y Microbiologia, Universidad Austral de Chile,

Casilla 567, Valdivia, Chile

* Departamento de Produccion Vegetal, Facultad de Agronomia, Universidad de Concepcion,

Av. Vicente Méndez 595, Chillan, Chile

* CORRESPONDENCE TO: xzhou@zafu.edu.cn

ABSTRACT—Neozygites linanensis sp. nov. was identified as infecting the aphid Takecallis

taiwanus collected on bamboo plants, Chimonobambusa quadrangularis, in the Lin‘an region

of southeast China during spring and summer 2014. This is the first record of a Neozygites

species on T: taiwanus. The Chinese Neozygites specimen is described, illustrated, and

discussed. The phylogenetic relationship among selected Neozygites species is evaluated

based on the successful sequence of the SSU rDNA gene from the new fungus.

KEY worpDs—natural enemies, entomopathogenic fungi, Entomophthoromycota, aphid

control, Neozygitaceae

Introduction

Bamboo plantations are a common forest type in Asia, and bamboo plants

supply food, ethnomedicines, and other necessary materials. Moreover,

bamboo forests are extremely helpful for carbon sequestration (Li et al. 2013).

Worldwide, over 40 aphid species infest bamboos, and many bamboo aphids

with robust reproductive and dispersal abilities cause considerable loss of

bamboo productivity (Blackman & Eastop 1994). Zhou et al. (2014) recently

reported on one entomophthoralean aphid pathogen, Conidiobolus obscurus

306 ... Zhou & al.

(I.M. Hall & PH. Dunn) Remaud. & S. Keller, that infected four bamboo aphid

species in the laboratory. However, rarely has information been recorded about

the natural enemies and pathogens of bamboo aphids and their interaction in

the field. Recently entomopathogenic fungi on bamboo aphids were surveyed

in China's Lin’an region, an important bamboo forest area. Mycosed individuals

of one widespread bamboo aphid species that frequently infects local bamboo

plants, Takecallis taiwanus (Blackman & Eastop 1994), were observed during

spring and early summer. Initial morphological examination of the infected

T. taiwanus showed that the entomopathogenic fungus represented Neozygites.

The only previously reported aphid-pathogenic Neozygites species in China is

Neozygites fresenii (Nowak.) Remaud. & S. Keller, which has been found on

Aphis gossypii in northern China and on Myzus persicae on cabbage in southern

China (Zhang et al. 1983, Huang et al. 2008).

Research on Neozygites species in natural ecosystems has been limited,

particularly in comparison to that reported worldwide for other aphid-specific

entomophthoralean species in Pandora, Conidiobolus, Entomophthora, and

Zoophthora (Barta & Cagan 2006, Keller 2006). Ben-Zeev (1981) reported

Neozygites turbinata (R.G. Kenneth) Remaud. & S. Keller in Israel on the aphid

Pterochloroides persicae. Neozygites lageniformis (Thaxt.) Remaud. & S. Keller

was found on aphids Myzocallis coryli on Corylus avellana in Chile (Barta

& Cagan 2006). In Switzerland, Keller reported Neozygites cinarae S. Keller

on Cinara pilicornis on Picea abies (Keller 1997) and described Neozygites

remaudierei S. Keller from Myzocallis coryli (Keller 2006). Barta (2009)

reported eighteen species of aphids on woody plants infected by N. fresenii

and one species infected by Neozygites cinarae in the Mlynany arboretum in

Slovakia. Montalva et al. (2013) discovered a new species, Neozygites osornensis

Montalva & Barta, on Cinara aphids associated with Cupressus macrocarpa

and Austrocedrus chilensis in Chile. Other aphid-pathogenic species reported

in agroecosystems include Neozygites microlophii S. Keller on nettle aphids

in Switzerland and Neozygites lecanii (Zimm.) Ben Zeev & R.G. Kenneth on

Lecanium viride in South East Asia.

Additionally, gene-based research on Neozygites species is scarce, which

further hinders understanding of the phylogenetic relationships and the status

of this genus within the entomophthoroid fungi. Finally, difficulty in obtaining

in vitro cultures and its infrequent collection makes Neozygites a much-

understudied genus (Humber 2012a). The aim of the present work was to identify

the Neozygites species collected on mycosed T: taiwanus populations in bamboo

forests through morphological and molecular analyses. For sequencing fungal

Neozygites linanensis sp. nov. (China) ... 307

DNA from resting spores within host cadavers, we conducted a subcloning

procedure that is widely used to diagnose mixed sequences in samples

(Pérez-Tris & Bensch 2005). We then used the sequence of the small subunit

ribosomal RNA (SSU rRNA) gene obtained from the mycosed T. taiwanus

specimens to analyze the genetic relationships of other Neozygites species.

Materials & methods

Field collection & material processing

The survey of local bamboo-Chimonobambusa quadrangularis forests in the suburb

of Liman city, Zhejiang province, China, was carried out during the spring and summer

of 2014. Specimens of dead mycosed adults of T: taiwanus attached to bamboo leaves

were carefully removed and transferred to a paper bag and then stored in a polystyrene

cooler at —20°C (Benjamin et al. 2004). The fungal pathogen (N51) was cultured in vivo

by host passage in leaf-containing Petri dishes in the laboratory at 20—24°C. Two types

of solid culture medium were tested during our attempts to let the fungal spores eject on

the surface of medium and obtain in vitro culture of this fungus: (i) Sabouraud dextrose

agar plus yeast extract (SDAY: 4% glucose, 1% peptone, 1% yeast extract, and 1.5% agar)

and (ii) SEMA (80% SDAY plus 11.5% egg yolk, 8.5% pure milk) were attempted to

cultivate the fungus (Hajek et al. 2012).

Morphological evaluations

The fungus was identified based on morphological characteristics according to

Keller (1991, 1997) and Humber (2012a). Semi-permanent slide mounts were prepared

in lactophenol-aceto-orcein (LPAO) as described by Keller (1987) and deposited in the

Forest Insect and Disease Herbarium, Zhejiang Agricultural and Forestry University,

Linan, China (ZAFU). Fungal structures were examined with a Zeiss AX10 microscope,

photographed with a Zeiss HAL100 digital camera at a magnification of 400x, and

measured with the software Zen 2012 (Zeiss). The measurements were based on 50

objects per microstructure. For each microstructure, mean value of measurements,

standard error of the mean (+ SEM), and maximal and minimal values were calculated,

cited below as minimum-mean+SEM-maximum.

The fungal material obtained from T. taiwanus in China was microscopically

compared with Neozygites fresenii attacking Rhopalosiphum padi in Poland (where the

fungus N. fresenii was first described) and N. fresenii from Slovakia, Austria (European

material was provided by Dr. Marek Barta), and Chile. Two other aphid-pathogenic

Neozygites species, N. osornensis and N. cinarae, were obtained during a survey of

natural enemies of tree dwelling aphids in southern Chile.

DNA extraction & PCR amplification

Resting spores of the Neozygites isolates for DNA extraction were obtained from

laboratory-infected T. taiwanus bamboo aphids. Fresh mycosed cadavers collected

from the field were placed in aphid cohorts in leaf-containing dishes at 20—24°C.

After three days of incubation, new cadavers, including black cadavers with resting

308 ... Zhou & al.

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Neozygites linanensis sp. nov. (China) ... 309

spores, were found in each dish. Cadavers with resting spores were collected, placed

into 1.5 mL tubes (50 cadavers per tube), and stored at 4°C. DNA extraction followed

the protocol set forth in the TakaRa (Ver. 5.0) MiniBest universal genomic DNA

extraction Kit. To maximize DNA extraction from the resting spores, several 2.5

mm sterile zirconia beads were added into each tube and then vibrated for 1 min,

repeated several times during the 56°C water bath for cell lysis. The extracted DNA

concentration was quantified by Nanodrop® ND-1000 and maintained at —20°C.

Universal fungal primers NSR 0004-5’ (5’-cTGGTTGATTCTGCCAGT-3’; Gargas &

DePriest 1996) and SR13 (5’-AATGATCCTTCCGCAGGT-3’; Depress 1993) were used to

amplify the nuclear SSU ribosomal RNA gene. PCR was carried out with 30 ul per

sample and in duplicates. Each sample contained 2 x PrimeSTAR® Max Premix, 0.12

uM of each primer, sterile distilled water, and 2 ul template (25 ng/ul of fungal DNA

isolated). The thermal profile of the 35 cycles of PCR consisted of the initial step at

94°C for 3 min, followed by 35 cycles at 94°C for 15 s, 55°C for 15 s and 72°C for 10

s and the final step at 72 °C for 5 min. PCR products were run in 1% agarose gels

with 0.1 ul/mL of ethidium bromide in 1 x TBE buffer at 100 V for 30 min. The PCR

products were examined and photographed on a UV light transilluminator.

DNA sequencing & analysis of SSU rDNA sequences

The amplified DNA was sent to TaKaRa Biotechnology (Dalian, China) for TA

cloning and sequencing. The target band of ~1.5 kb was recovered, and the DNA

fragment was ligated into a pMD18-T vector (TaKaRa, China) after adenine additions.

Escherichia coli JM109 cells were transformed and spread onto plates of Luria-Bertani

agar plus X-Gal, IPTG and Ampicillin. The plates were incubated overnight at 37°C.

Three positive colonies (white color) were selected through blue-white screening

as replicates. The vector primers M13-47 (5’-CGCCAGGGTTTT CCCAGTCACGAC-3’)

and RV-M (5’-GAGCGGATAACAATTTCACACAGG-3’) were used to amplify the SSU

rRNA gene sequence of the Neozygites isolate, which was inserted into the pMD18-T

plasmid in monoclonal colony PCR. And then the PCR products from each clone

were sequenced. Copies of the Neozygites sp. (N51) sequences were pairwise compared

with other Neozygites species online (ClustalW2, the European Bioinformatics

Institute, Cambridge, UK). A multiple alignment of sequences was performed with

ClustalX 2.0 (Larkin et al., 2007) among SSU rDNA sequences, and a dendrogram was

constructed with Seaview 4.0 (Gouy et al. 2010) using the neighbor-joining method

with 1000 bootstraps. The sequences obtained in this analysis and from GenBank for

other Neozygites species such as N. osornensis (KC822922), N. cinarae (KC822923),

N. turbinata (KC822924), N. parvispora (MacLeod & Carl) Remaud. & S. Keller

(AF296760), N. floridana (J. Weiser & Muma) Remaud. & S. Keller (AF296758),

N. tanajoae Delal. et al. (AY233981), and Neozygites sp. (AF296759) obtained from

GenBank were included in the analysis. Other species infecting aphids on bamboo

plants as Conidiobolus obscurus (JQ014014) (Zhou et al. 2014) were chosen as outgroup

in the phylogenetic tree.

310... Zhou & al.

Results

During the survey of natural enemies of the bamboo aphid, mycosed

cadavers of T: taiwanus were observed from spring to summer 2014. Grey-

brown infected aphids were easily recognized hanging on the lower sides of

new bamboo leaves. Microscopic observations of the cadavers revealing fungal

infection by Neozygites represented the first find of mycosed bamboo aphids in

Lin/an city, southeast China.

Cultural & morphological observations

No in vitro cultures were successfully isolated from infected individuals,

despite attempts to isolate the entomopathogenic fungi using two different

solid culture media.

Measurements of taxonomically important structures and comparison

with morphologically similar species are presented in TABLE 1. Neozygites

obtained from T. taiwanus in China was morphologically distinguished from

the other aphid Neozygites species mainly by higher number of nuclei per

hyphal body, primary conidia, and capilliconidia (PLATE 1). The sizes of the

major morphological microstructures in the Chinese fungus are larger than in

N. fresenii from Austria and Slovakia, similar in N. fresenii from Poland and

Chile, and smaller than in N. osornensis and N. cinarae from Chile (except for a

larger capilliconidial diameter in the Chinese Neozygites specimens).

Molecular analysis

The fungal SSU rDNA gene from the Chinese Neozygites fungus was

successfully sequenced and its sequence (KM386989) deposited in the GenBank

database (NCBI). Pairwise comparisons of the 1524 bp long aligned SSU rDNA

gene sequence showed that the Chinese fungal sequence shares a 92% similarity

with N. turbinata and a 93% similarity with N. cinarae and N. osornensis—

all species infecting aphid hosts. On the other hand, the Chinese Neozygites

sequence matched only 73% with N. tanajoae, 75% with N. floridana, and 77%

with N. parvispora, all species affecting only mites or thrips. Furthermore, the

phylogenetic neighbor-joining SSU rDNA tree separates the bamboo aphid

fungus from the clade containing other aphid-associated Neozygites species,

supporting the fungus found on T! taiwanus as an independent species within

Neozygites (PLATE 2).

Taxonomic description

Neozygites linanesis X. Zhou & C. Montalva, sp. nov. PLATE 1

MycoBank 814659

Differs from Neozygites fresenii by its larger hyphal bodies and primary conidia (both

containing a greater number of nuclei) and by its smaller zygospores.

Neozygites linanensis sp. nov. (China) ... 311

PLaTE 1. Fungal structures of Neozygites linanesis on bamboo aphid Takecallis taiwanus.

a. Unbranched conidiophores. b. Subspherical hyphal body with visible nuclei. c. Maturing

resting spores filling cadaver. d. Resting spores with two visible nuclei. e. Primary conidium.

f. A developing capilliconidium on a long slender capillary tube. g. Mature capilliconidia adhered

to host leg parts. h. Resting spore developing by conjugation of two spherical hyphal bodies.

[Figures a, b, d, e, h stained in lactophenol-aceto-orcein; figures c, f, g in cotton blue.]

Type: China, Zhejiang Province, suburb of Lin’an city, 30°15’49”N 119°44’23”E, from

Takecallis taiwanus (Takakashi) on Chimonobambusa quadrangularis (Fenzl) Makino,

20 May 2014, coll. Xiang Zhou (Holotype, ZAFU IP15091101; GenBank KM386989),

ErymMo.oey: The specific epithet refers to the name of the type locality.

Hyphal bodies spherical to subspherical, 14.5-17.4+1.6-21 um in diameter,

containing an average of 10 nuclei (1.9 + 0.6 um diam.); primary conidia

spherical, 18.8-22.9+1.8-26.5 x 14.4-17.6+1.9-22.8 um with a distinct cylindrical

papilla, containing an average of 7 nuclei; capilliconidia forming from primary

conidia, almond-shaped, 17.3-23.3+2.3-29.3 x 10.6-13.6+1.6-16.2 um, containing

an average of 7 nuclei, borne on a long slender capillary tube 66.5 + 15.8 um;

zygospores broadly ellipsoidal, thick-walled, with a black, smooth episporium,

21.6-28.1+3.7-37.2 x 15.5-19.2+2.3-26.9 um. Rhizoids not observed.

Observed in spring and summer 2014 in just one locality in China.

Discussion

The genus Neozygites, which differs from the other entomophthoroid

fungal species by the nuclear structure and behaviour during mitosis (Butt &

Humber 1989), currently comprises 19 entomopathogenic fungal species that

312 ... Zhou &al.

may eventually be split into two genera differentiated by their host affinities

and resting spore morphologies (Humber 2012b). The nine aphid-pathogenic

species (Keller 2006, Montalva et al. 2014a) have smooth ovoid resting spores,

and none of these species (including the type species, N. fresenii) has been grown

in vitro; the remaining 10 species infect primarily mites (but include some

species on non-aphid insect hosts) and produce rough globose zygospores; all

in vitro cultures of Neozygites species have been isolated from mites.

From the morphological point of view, N. fresenii and N. linanesis are closely

related. Neozygites fresenii is a widely distributed fungal pathogen of aphids

(Milner & Holdom 1986, Hatting et al. 1999, Barta & Cagan 2006, Scorsetti

et al. 2007). It was named and described as Empusa fresenii by Nowakowski

(1883), who first discovered the fungus in naturally infected aphids collected

in Poland. This is the first described member of the Neozygites genus. While

both fungi are pathogenic to aphids, N. linanesis can be distinguished from

N. fresenii mainly by its greater average number of nuclei per hyphal body and

primary conidium. Also, the hyphal bodies and primary spores in N. linanesis

are somewhat larger than those in N. fresenii recorded in the literature from

Poland, Switzerland, and Slovakia, while its zygospores are slightly small

compared those recorded for N. fresenii (Keller 1991, Batazy 1993, Barta 2004).

Compared to other aphid-pathogenic Neozygites species, all the recorded

dimensions for microstructures in N. microlophii and N. remaudierei are larger

than those in N. linanesis (Keller 1991, 2006).

Unfortunately, there are no DNA sequences from N. fresenii available for

molecular comparisons with other Neozygites species. Nonetheless, SSU rDNA

gene analysis from N. linanesis and the other aphid-pathogenic Neozygites

species showed clearly that N. linanesis is separated in a different clade from

N. cinarae, N. turbinata, and N. osornensis (PLATE 2); this analysis, when

combined with the morphological differences, suggests that N. linanesis is a

distinct species within Neozygites.

Moreover, the molecular differences between aphid- and mite-/thrip-

pathogenic Neozygites species are consistent with the proposal by Humber

(2012b). SSU rDNA gene-based analysis successfully separated N. linanesis

from a morphologically similar Neozygites species, thus supporting the

morphological observations made in this study. This gene has been previously

used to separate N. tanajoae from the morphologically similar N. floridana

(Delalibera & Humber 2004) as well as N. osornensis from N. turbinata and

N. cinarae (Montalva et al. 2014b). We regard the SSU rDNA gene as a valid

tool for identifying Neozygites species.

Neozygites linanensis sp. nov. (China) ... 313

N. linanensis (KM386989)

100 N. cinarae (KC822923)

199 || NV. turbinata (KC822924)

splude Buizyisesed

N. asornensis (KC822922) J

N. parvispora (AF296760) j

Neozygites sp. (AF296759)

92 [

100 N. tanajoae (AY233981)

N. floridana (AF296758)

sdiiy} pue sayiw suizuisesed

C. obscurus (JQ014014)

0.01

PLATE 2. Dendrogram showing the relationships among SSU rDNA sequences of Neozygites

linanesis isolated from bamboo aphid Takecallis taiwanus and other DNA sequences of Neozygites

species published in GenBank. Conidiobolus obscurus was used as outgroup and the numbers on

each branch indicate bootstrap values (1000 replicates).

In this work we have described a new Neozygites species that can infect

Takecallis aphids and observed the spread of N. linanensis in host populations

during the aphid infestation period. Neozygites linanesis, which appears to be

a natural control agent of T’ taiwanus in bamboo forests in China, represents

the first record of a Neozygites species associated with T! taiwanus populations.

Our 2014 surveys also revealed that N. linanesis infects other aphid species

in China, such as Metamacropodaphis bambusisucta. The identification of

N. linanesis and the sequencing of its SSU rDNA gene have thereby added to

the knowledge regarding natural fungal enemies of bamboo aphids.

Acknowledgments

The authors are indebted to Marek Barta (Mlynany Arboretum SAS, Slovakia) for the

help on morphological identification of fungi and Xianyu Yang (Zhejiang Agricultural

and Forest University, China) for the valued suggestion on molecular manipulation.

They also thank Marek Barta and Ming-Guang Feng (Zhejiang University, China) for

314 ... Zhou &al.

presubmission review and Richard Humber (United States Department of Agriculture,

Agricultural Research Service, New York) and Christian Luz (Instituto de Patologia

Tropical e Saude Publica, Universidade Federal de Goias, Brasil) for additional critical

review of the manuscript. This study was supported by the grants from the Natural

Science Foundation of China (31300550, 31200487).

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MYCOTAXON

April-June 2017— Volume 132, pp. 317-326

https://doi.org/10.5248/132.317

Five Lecidea lichens new to China

XIANG-XIANG ZHAO “, ZUN-TIAN ZHAO **,

ConG-ConcG MIAO ', ZHAO-JIE REN? & LU-LU ZHANG ©

' Key Laboratory of Plant Stress Research, College of Life Sciences,

Shandong Normal University, Jinan, 250014, PR. China

? Shandong Provincial Museum, Jinan, 250014, P. R. China

“ CORRESPONDENCE TO: 18264101801@163.com

ABSTRACT—Five Lecidea lichen taxa—L. andersonii, L. grisella, L. laboriosa, L. atrobrunnea

subsp. saxosa, L. atrobrunnea subsp. stictica—are reported for the first time from China.

Keyworps—Asia, Lecidiaceae, Lecideales, saxicolous lichens, taxonomy

Introduction

The lichen genus Lecidea Ach. (Lecideaceae) was originally described by

Acharius (1803). In the sense of Zahlbruckner, the genus once represented

one of the largest lichen genera and included about 1200 species (Schmull et

al. 2011). Of the 427 species included in Lecidea sensu lato (Kirk et al. 2008),

only about 100 (all saxicolous) are accepted in Lecidea sensu stricto, which is

characterized by a Lecidea-type ascus (Hertel 2006, Smith et al. 2009).

During our study of lecideoid lichens from China, five Lecidea taxa were

recognized as new to the country—L. andersonii, L. grisella, L. laboriosa,

L. atrobrunnea subsp. saxosa, and L. atrobrunnea subsp. stictica.

Materials & methods

The specimens studied are preserved in either the Lichen Section of Botanical

Herbarium, Shandong Normal University, Jinan, China (SDNU) or the Kunming

Institute of Botany, Chinese Academy of Sciences, Kunming, China (KUN). The

* Xiang-Xiang Zhao & Zun-Tian Zhao contributed equally to this research.

318 ... Zhao, Zhao & al.

specimens were examined using standard microscopic techniques and hand-

sectioned under a Nikon SMZ 645 dissecting microscope. Anatomical descriptions

were based on observations of these preparations under a Nikon EclipseE200

microscope. Sizes cited for the thallus, apothecium, hymenium, and exciple are

based on five measurements per specimen. Ascospore dimensions are based on ten

measurements per specimen and presented as a range, with outlying values given

in parentheses. Secondary metabolites of all the specimens were identified using

TLC and solvent C (Orange et al. 2010). Medullas were tested for amyloid reactions

using IKI (10% aqueous potassium iodide). Photos of these lichens were taken under

Olympus SZX16 and BX61 with DP72.

Taxonomic descriptions

Lecidea andersonii Filson, Muelleria 3(1): 16 (1974). Fic. 1

THALLUS often poorly developed, rimose to areolate, angular and usually

appressed along rock crevices, whitish to grey; medulla I+ intensively violet-

blue; prothallus absent. APOTHECIA sessile, constricted at the base, round to

irregular, (0.4-)1-1.4 mm diam.; disc black, flat to slightly convex, mostly

epruinose, rarely with a thin whitish pruina; margin distinct, sometimes

becoming flexuose. Exciple hyaline, outer margin dark green to brownish

green, 90-150 um wide; epihymenium dark green; hymenium hyaline, 50-80

uum high; hypothecium brownish, with crystals; paraphyses simple, coherent,

with swollen apices. Asci clavate; 8-spored; ascospores narrowly ellipsoid,

(8—)9-12 x 3.5-4.4 um. Pycnrp1A not observed.

CHEMISTRY—Cortex and medulla K-, C-, KC-. 4-O-demethylplanaic acid

was detected by TLC.

SUBSTRATE—On siliceous rocks.

DIsTRIBUTION—Lecidea andersonii has been reported from Europe and

Antarctica (Hertel et al. 2009, Ruprecht et al. 2010). New to China.

SPECIMENS EXAMINED: CHINA. XIZANG, Anduo, Mt. Tanggula, alt. 5200 m, on rock, 30

Jul. 2011, Y.L. Cheng 20118836A, 20116422, 20116396, 20116343, 20116395, 20126439

(SDNU); XINJIANG Bukang, Mt. Tianshan, alt. 3700 m, on rock, 17 Aug. 2012, L.L.

Zhang 20129086, 20129092, 20129135 (SDNU); SicHuAN, Litang, Mt. Kazila, alt. 4700

m, on rock, 7 Nov. 2008, Z.J. Ren 20080005, 20108491, 20080159 (SDNU).

ComMMENTS—Lecidea andersonii is morphologically similar to L. diducens,

also with an endolithic thallus, sessile apothecia, dark green epihymenium,

brownish hypothecium, and narrowly ellipsoid ascospores, but L. diducens

is distinguished by an I- medulla and C+ red exciple and contains

2’-O-methylanziaic acid (Inoue 1991).

Lecidea spp. new to China... 319

(a.

wt:

ree aw

a =

ett ee a

; H / SS

a ey)

/ ave }} SS

b ie

yy ts

eae w

¥ J F, n ~~ \,

\ ry 7 \ =)

. + » ' , a

q * ‘

h — SS

Mee Odd =

E a a; ' La ’

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(t re WAS ne

Be tre | ed 10um | 10 pm

Fic. 1. Lecidea andersonii (20129135, SDNU). A: thallus; B: apothecia; C: apothecium section;

D: exciple and hypothecium with crystals; E: ascus and ascospores; F: amyloid reaction of ascus;

G: paraphyses; H: ascospores.

Lecidea grisella Flérke, Lich. Schlesien: nos 141-142 (1829). Fig. 2

THALLUS contiguous, cracked, rimose, rarely areolate, whitish grey to pale

grey-brown, generally thicker and never convex; medulla I-; prothallus absent.

APOTHECIA black, immersed, <0.6-1.2(-1.5) mm diam.., flat to slightly concave;

margin distinct, regular. Exciple dark brown, with crystals; epihymenium olive-

320 ... Zhao, Zhao & al.

Fic. 2. Lecidea grisella (YN182, SDNU). A: thallus; B: apothecium section; C: exciple and

hypothecium with crystals; D: paraphyses; E: amyloid reaction of ascus; F: ascus and ascospores;

G: ascospores.

brown, with little crystals; hymenium colorless, 60-80 um; hypothecium dark

brown, without crystals; paraphyses 2-3.5 um thick. Asci clavate; ascospores

ellipsoid, simple, 8-12(-13) x 5-6(-7) um. Pycnrp1A not observed.

CHEMISTRY—Medulla K-, C+ red, KC+ red; exciple C+ red. Chemical

constituent: gyrophoric acid.

Lecidea spp. new to China... 321

SUBSTRATE—On siliceous rocks.

DISTRIBUTION—Lecidea grisella has been reported from Asia, Africa, and

Europe (Smith et al. 2009). New to China.

SPECIMENS EXAMINED: CHINA. YUNNAN, Tangdan, Lanniping, alt. 3800 m, on rocks, 14

May. 2014, W.C. Wang YN222, YN227, YN226, YN225, YN252; L. Hu YN185, YN181,

YN182, YN183 (SDNU).

ComMENts —Lecidea grisella is similar to L. fuscoatra. The main difference

between the two species is visible at the margins: L. fuscoatra forms as an

areolated thallus on a prothallus, whereas L. grisella forms with a continuous

thallus that becomes rimose or splits secondarily up into areoles that tend to

remain flat with angular edges for a long time (Aptroot & Herk 2007). Lecidea

fuscoatra occurs on a wider variety of substrata (rather smooth, slightly

nutrient-rich siliceous rocks, walls, and brick) and, unlike L. grisella, can be

found growing on roof tiles and (rarely) on timber (Smith et al. 2009).

Lecidea laboriosa Mill. Arg., Flora 57: 187 (1874). Fic. 3

THALLUs endolithic, rarely epilithic; medulla white, I-; prothallus absent.

APOTHECIA black, loosely attached with strongly constricted base, 0.6-1.4

mm diam; disc black, flat to convex, epruinose or faintly pruinose; margin

black, persistent, regular to flexuose. Exciple unpigmented or sometimes pale

brown, peripherally dark brown, 30-80 um wide, with crystals; epihymenium

black green, olive or brown; hymenium colorless or more rarely with a pale

violet tinge, 37.5-60 um tall, I+ blue; subhymenium colorless, 25-50 um thick;

hypothecium colorless to pale brown, with crystals; paraphyses simple, rarely

branched, anastomosed, apical cells 3.5-5 um wide, globose, with dark caps.

Asci clavate, 8-spored; ascospores oblong, 7-11.5 x 2.5-4 um. PYcNIDIA not

observed.

CHEMISTRY—Cortex and medulla K-, C-, KC-. 4-O-demethylplanaic acid

was detected by TLC.

SUBSTRATE—On acidic rocks.

DisTRIBUTION—Lecidea laboriosa has been reported from Asia, Europe,

and North and South America (Hertel et al. 2009, Nash et al. 2004). New to

China.

SPECIMENS EXAMINED: CHINA. XINJIANG, Ursula, Bayingou, alt. 2100 m, on rock, 23

Jul. 2013, C. Chao XL0239 (SDNU); Aletai, Buerjin, Bulanai, alt. 1920 m, on rock, 23 Jul.

2013, Q. Ren 2012607 (SDNU)); Yili state, Qiongbola Qapqal national Forest Park, alt.

1861 m, on rock, 12 Jun. 2014, PM. Wang 20140152 (SDNU); StcHuan, Daocheng, Mt.

Haizi, alt. 4390 m, on rock, 19 Sep. 2002, L.S. Wang 02-21602 (KUN).

322 ... Zhao, Zhao & al.

Fic. 3. Lecidea laboriosa (2012607, SDNU). A: thallus; B: apothecia; C: apothecium section;

D: exciple and hypothecium with crystals; E: ascus and ascospores; F: amyloid reaction of ascus;

G: ascospores.

ComMENtTS—Lecidea laboriosa is morphologically similar to L. auriculata,

which is distinguished by a K+ red-brown exciple, I+ violet-blue medulla, and

the presence of confluentic acid (Nash et al. 2004).

Lecidea atrobrunnea subsp. saxosa Hertel & Leuckert,

Lich. Flora Gr. Sonoran Desert Region 2: 296 (2004). Fic. 4

Lecidea spp. new to China... 323

RUSS Nise) 10 um i)

SOON eo Oe a

10 um

Fic. 4. Lecidea atrobrunnea subsp. saxosa (20140920, SDNU). A: thallus and apothecia;

B: apothecium section; C: exciple and hypothecium with crystals; D: paraphyses; E: amyloid

reaction of ascus; F: ascus and ascospores; G: ascospores.

THALLUS crustose, bullate to subsquamulose, continuous, brown to dark

brown, polished, with an epinecral layer, 5-30 um, areolate; medulla I+ violet-

blue, hypothallus black. AporHecta black, epruinose, <1.6 mm diam., adnate,

between or partly on the areolae, moderately constricted at the base; with a

somewhat prominent margin. Exciple with a greenish black, epihymenium-

like rim, unpigmented interior; epihymenium green or olive-green; hymenium

324 ... Zhao, Zhao & al.

40-60 um tall; subhymenium hyaline 15-40 um tall; hypothecium hyaline

to pale ochre; paraphyses simple, anastomosing, with swollen apices.

Asci clavate, 8-spored; ascospores hyaline, ellipsoid-oblong, simple, 7-10 x

3.5-4 um. Pycnip1iA not observed.

CHEMISTRY—Cortex and medulla K+ red, C-, KC-. Norstictic acid was

detected by TLC.

SUBSTRATE—on Siliceous rocks.

DIsTRIBUTION—Lecidea atrobrunnea subsp. saxosa has been reported from

Asia, Europe, North America, and Antarctica (Nash et al. 2004). New to China.

SPECIMENS EXAMINED: CHINA. XINJIANG, Gongnaisi, Panchen Ditch, alt. 3800 m, on

rock, 3 Jul.2014, PM. Wang 20140675A (SDNU); Hucheng, Kurt, alt. 3700 m, on rock,

12 Jul.2014, PM. Wang 20140950, 20140920 (SDNU).

CoMMENTS—Lecidea atrobrunnea subsp. saxosa is similar to L. atrobrunnea

subsp. atrobrunnea; but subsp. saxosa contains norstictic acid while subsp.

atrobrunnea contains confluentic acid (Nash et al. 2004).

Lecidea atrobrunnea subsp. stictica Hertel & Leuckert,

Lich. Flora Gr. Sonoran Desert Region 2: 297 (2004). Fic. 5

THALLUS crustose, bullate to subsquamulose, continuous, brown to dark

brown, polished, with an epinecral layer, 5-30 um, areolate; medulla I+

violet-blue, hypothallus black. ApotHecia black, epruinose, <1.7(-2.5) mm

diam., adnate, between or partly on the areolae, moderately constricted at

the base; with somewhat prominent margin. Exciple with a greenish black,

epihymenium-like rim, unpigmented inside; epihymenium bright green,

olive-green, to green-brown; hymenium 40-60 um tall; subhymenium hyaline

15-60 um tall; hypothecium hyaline to medium brown (rarely dark brown);

paraphyses simple, anastomosing, with swollen apices. Asct clavate, 8-spored;

ascospores hyaline, ellipsoid-oblong, simple, 7-11 x 3-4.5 um. PycNipIA not

observed.

CHEMISTRY—Cortex and medulla K-, C-, KC-. Stictic acid and hypostictic

acid was detected by TLC.

SUBSTRATE—On siliceous rocks.

DIsTRIBUTION—Lecidea atrobrunnea subsp. stictica has been reported from

Asia, Europe, North America, and Antarctica (Nash et al. 2004). New to China.

SPECIMENS EXAMINED: CHINA. XINJIANG, Hami, Baishitou, alt. 2800 m, on rock, 28

Jul. 2013, C.Li XLO111 (SDNU); Gongnai, Panchen Ditch, alt. 3800 m, on rock, 3 Jul.

2014, PM. Wang 20140675B (SDNU).

CoMMENTS—Lecidea atrobrunnea subsp. stictica is morphologically similar

to L. atrobrunnea subsp. atrobrunnea and L. atrobrunnea subsp. saxosa, but

subsp. stictica contains stictic acid or hypostictic acid (Nash et al. 2004).

Lecidea spp. new to China... 325

10 um

Fic. 5. Lecidea atrobrunnea subsp. stictica (20140675B, SDNU). A: thallus and apothecia;

B: apothecium section; C: exciple with crystals; D: paraphyses; E: amyloid reaction of ascus;

F: ascus and ascospores; G: ascospores.

Acknowledgements

We thank Dr. G. Rambold (University of Bayreuth, Bayreuth, Germany) and

Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing, China) for presubmission reviews. And we

thank Dr. Alan M. Fryday (Herbarium, Department of Plant Biology, Michigan State

University) for providing great help during our research. This work was supported

326 ... Zhao, Zhao & al.

by the National Natural Science Foundation of China (31400015, 31570017),

and the Scientific Research Foundation of Graduate School of Shandong Normal

University (SCX201629).

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Acharius E. 1803. Methodus qua omnes detectos Lichenes, vols 1 & 2. Stockholm. LV + 394 p.

Aptroot A, Herk CM van. 2007. Lecidea grisella sympatric with Lecidea fuscoatra, differing in its

rimose instead of areolate thallus. Lichenologist 39(3): 293-296.

https://doi.org/10.1017/S0024282907006706

Hertel H. 2006. World distribution of species of Lecidea (Lecanorales) occurring in Central

Europe.19-74, in: A Lackovieova et al. (eds). Central European Lichens — Diversity and

Threat. Mycotaxon Ltd., Ithaca, New York.

Hertel H, Aptroot A, Seaward MRD, Sparrius LB. 2009. A new key to cryptothalline species of the

genus Lecidea (Lecanorales). Bibliotheca Lichenologica 99: 185-204.

Inoue M. 1991. Lecideoid lichens of Prince Olav Coast and Séya Coast, Enderby Land, East

Antarctica. NankyokuShiry6 35(3): 271-284.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th edition.

CABI Bioscience: CAB International. 711 p.

Nash TH III, Ryan BD, Diederich P, Gries C, Bungartz F (eds). 2004. Lichen flora of the greater

Sonoran Desert Region, vol. 2. Lichens Unlimited, Arizona State University, Tempe, Arizona.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

Ruprecht U, Lumbsch HT, Brunauer G, Green TGA, Turk R. 2010. Diversity of Lecidea (Lecideaceae,

Ascomycota) species revealed by molecular data and morphological characters. Antarctic

Science 22(6): 727-741. https://doi.org/10.1017/S0954102010000477

Schmull M, Miadlikowska J, Pelzer M, Stocker-Wo6rgotter E, Hofstetter V, Fraker E, Hodkinson

BP, Reeb V, Kukwa M, Lumbsch HT, Kauff F, Lutzoni F. 2011. Phylogenetic affiliations of

members of the heterogeneous lichen-forming fungi of the genus Lecidea sensu Zahlbruckner

(Lecanoromycetes, Ascomycota). Mycologia 103(5): 983-1003. https://doi.org/10.3852/10-234

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds.). 2009.

The lichens of Great Britain and Ireland. Natural History Museum Publications, in association

with The British Lichen Society.

MY COTAXON

April-June 2017— Volume 132, pp. 327-335

https://doi.org/10.5248/132.327

Pseudobaeospora lilacina sp. nov.,

the first report of the genus from China

SHUANG- YAN WU |, JIA-JIA L1?, MING ZHANG ?,

XIAO-DAN Yu **, SHU-XIA Lv * & DONG-XU Cao *

‘College of Biological Science and Technology, Shenyang Agricultural University,

Shenyang, Liaoning 110866, China,

? School of Bioscience & Bioengineering, South China University of Technology,

Guangzhou 510006, China

* CORRESPONDENCE TO: yuxd126@126.com

ABSTRACT—A new species, Pseudobaeospora lilacina, is described based on materials

collected from Guangdong Province, China. The fungus represents the first record of the

genus in China. The new species is characterized by its lilac pileus, stubbly stipe, and ellipsoid

spores. Phylogenetic analyses of the internal transcribed spacer regions (ITS) and nuclear

ribosomal RNA gene large subunit (nrLSU) support P lilacina within Pseudobaeospora and

as distinct from all other sequenced species of the genus. The combination of morphological

and molecular data confirmed the fungus to be new. The new species is compared to

morphologically similar species and its taxonomic position is discussed.

Key worps —Agaricales, Basidiomycota, taxonomy, Tricholomataceae

Introduction

Pseudobaeospora Singer was erected by Singer (1942) and emended by Bas

(2003); the genus is characterized by small basidiocarps, pale to dark lilac pilei,

and small spores. Bas (2003), who described 25 Pseudobaeospora species, divided

the genus into five groups (Albidula, Celluloderma, Pyrifer, Frieslandica, and

Pillodii) based primarily on basidiocarp color and the presence or absence of

clamp connections. Kirk et al. (2008) estimated about 20 species for the genus,

while the current Index Fungorum database (http://www.index fungorum.org

accessed June 2016) lists 34 published taxa.

328 ... Wu &al.

The type species of the genus, Pseudobaeospora oligophylla (Singer) Singer,

was originally found in the Altai Mountains of central Asia by Singer (1938; as

Baeospora oligophylla). Pseudobaeospora species were subsequently described

from Asia, Europe, and North and South America (Singer 1986). Additional

species have been described from California (Vellinga 2001, 2009; Schwarz

2012), Hawaii (Desjardin et al. 2014), and Slovakia (Adam¢cik & Jancovicova

2011), but the genus remained unreported from China.

The taxonomic position of Pseudobaeospora has been ambiguous. Singer

(1942) initially placed the genus in Tricholomataceae as an independent genus

but soon afterwards transferred it to Agaricaceae, at first in tribus Lepioteae

(Singer 1951) and later in tribus Cystodermateae (Singer 1986). However,

Kuhner (1980) retained the genus in Tricholomataceae, a placement supported

by subsequent researchers (Bas 2003, Desjardin et al. 2014).

Here we describe a new species of Pseudobaeospora from China based on

a combination of morphological and molecular data; we also compare the

morphological similarities and differences between the new P lilacina and

previously described Pseudobaeospora species.

Material & methods

Fungal collections

Fungal specimens were collected from Guangdong Province, China. Tissue

blocks were removed from the inner part of fresh basidiomata for DNA analyses. The

specimens were dried with an electric air-ventilation drier and deposited in either the

Fungal Herbarium of Shenyang Agricultural University, Shenyang, China (SYAU),

or the Mycological Herbarium of Guangdong Institute of Microbiology, Guangzhou,

China (GDGM).

Morphological observations

The macroscopic description is based on the fresh specimens. The names of colors

are based on Ridgway (1912). Microscopic observations were made on sections of dried

specimens, using a 5% KOH solution and in Melzer’s reagent, using a light microscope.

The Q-value (quotient of length/width) for each spore (20 spores for each specimen)

was calculated, and the mean values were used in the descriptions.

For observation of the surface of spores, the gills were coated with gold. The

spores were examined with a scanning electron microscope (SEM; Zeiss & Ultra Plus,

Germany).

DNA extraction, amplification, and sequencing

Genomic DNA was extracted from the fresh blocks of tissue of the specimens using

the Plant DNA Extraction Kit (Sunbiotech Co., Ltd, Beijing). The raw DNA extracts

were used as templates for PCR. Amplification primers were ITS5/ITS4 (White et al.

Pseudobaeospora lilacina sp. nov. (China) ... 329

1990) for the ITS1+5.8S+ITS2 region and LROR/LR7 (http://www.biology.duke.edu/

fungi/mycolab/primers.htm) for the 5¢ end of 28S rRNA gene. Both reaction mixtures

and PCR conditions followed those in Yu et al. (2014). The PCR products were checked

on a 1% agarose gel stained with ethidium bromide. The DNA bands were visualized on

a UV transilluminator and documented digitally using the BIO-RAD ChemiDoc XRS

imaging system. Sequencing was performed on an ABI Prism’ 3730 Genetic Analyzer

(PE Applied Biosystems, USA).

DNA sequence analyses

The BLAST database (Altschul et al. 1997) was searched using the ITS and nrLSU

sequences from the type collection (SYAU-FUNGI-009) as the queries (23 May

2016). In order to confirm the taxonomic position of the new species, the sequences

of the Tricholomatoid clade (Matheny et al. 2006) were also retrieved from GenBank.

Nucleotide sequences amplified from the collections of the new species were aligned

with 44 ITS and 51 LSU sequences retrieved from GenBank using BioEdit 5.0.6 (Hall

1999) and Clustal X (Thompson et al. 1997). Based on the results of Matheny et al.

(2006), the sequences of Agaricus bisporus (J.E. Lange) Imbach (DQ404388 for ITS;

AY 635775 for nrLSU) were downloaded and included for rooting purposes.

A few dozen bases at both ends of the sequences were excluded from the analyses

because of the uncertainty in the base cells of the sequences. Finally, the data matrices

with 689 bp for ITS and 955 bp for nrLSU were produced. Nodal bootstrap support

(BS) was assessed with nonparametric bootstrapping using 1000 replicates. Bayesian

analysis was conducted using MrBayes v.3.1.2 (Ronquist & Huelsenbeck 2003).

The best-fitting sequence evolution model was chosen using MrModelTest v. 2.2

(Nylander 2004). Bayesian analyses of the ITS and nrLSU regions were run for 2 000

000 generations, all under the GTR model, with four chains, and trees sampled every

100 generations. The average split frequencies were checked to determine optimal

convergence of the chains below 0.01 after each generation. The first 25% (1250 trees)

of the sample trees was designated as burn-in, with the remaining samples retained for

further analyses. The topologies were used to generate a 50% majority rule consensus

tree for Posterior Probabilities (PP). Alignments have been deposited in TreeBASE

(http://purl.org/phylo/treebase/phylows/study/TB2:S19825 and $19826).

Results

Four ITS sequences and four nrLSU sequences were obtained from fresh

basidiomata. The average standard deviation of split frequencies in the

Bayesian analysis was less than 0.01 after 2,000,000 generations. Both trees

were rooted with Agaricus bisporus (FIGS. 1, 2). The ITS and nrLSU sequence

analyses both placed the genus Pseudobaeospora in a monophyletic group (PP

= 1) sister to Callistosporium graminicolor Lennox (Tricholomataceae). The new

species, P. lilacina, grouped with the other Pseudobaeospora species (PB. deckeri

C.F. Schwarz, P. pyrifera Bas & L.G. Krieglst., P wipapatiae Desjardin et al.) but

formed a separate branch with strong support (PP = 1).

330 ... Wu &al.

Pseudobacospora lilacina KX266951

Pseudobacospora lilacina KUS28842

| Psewdobacospora lilacina KU528840 [T]

Pseudobaeospora lilacina KU528841

arf Pseudobaeospora pyrifera AF391034

; | | Pseudobaeospora deckeri JF898319

os? Pseudobaeospora wipapatiae KF27179&

Callistosporium sp. DQ484065

Lyophylium leucophaeatum AF357032

a Calocybe carnea AF357028

1{_ Noelanea sericea DQ367430

Nolanea strictior DQ494680

1 [ Entoloma prunuloides DQ206983

Entoloma sinuatum AY691891

1 Lyophyllunt sp. DQ182502

Lyophyllum decastes AF3570S9

ow Clitocybe candicans DQ202268

| | 0.85

099 Ossicaulis lignatilis DQ825426

ca | Termitomyeces sp. DQ494698

092 Tricholomella constricta DQ825429

096 Dendrocollybia racemosa DQ825425

Asterophora lycoperdoides AF357037

Clitocybe subditopoda DQ202269

097 "Inocephalus sp. DQ490636

“= Rhodocybe mundula DQ494694

0.99

Neohygrophorus angelesianus DQ494678

* Cyathus striatus DQ486697

Clitocybe nebularis DQ486691

dss[ Collybia tuberosa AY854072

Lepista irina DQ221109

09s 099 Tricholoma inamoenum AF377246

ost Tricholoma matsutake AB138557

Tricholoma saponaceunt DQ494700

Tricholoma myomyces| DQ825428

Tricholoma myomyces2 AF377210

Mycena plumbea DQ494677

Mycena galericulatal DQ404392

Mycena amicta DQ490645

Panellus stypticus AB084488

Catathelasma ventricosum DQ486686

Agaricus bisporus DQ404388

0.07

Fic. 1. Fifty percent majority-rule Bayesian cladogram of Pseudobaeospora and related taxa based

on ITS sequence analysis. Node support is indicated by Bayesian posterior probabilities >0.5 on

branches. Pseudobaeospora lilacina sequences are presented in boldface.

Pseudobaeospora lilacina sp. nov. (China) ... 331

Entoloma sinuatum AY691891

Entoloma prunuloides AY700180

Pseudobaeospora lilacina KX266952

! Pseudobaeospora lilacina KU528836 [T]

Pseudobaeospora lilacina KU528838

a Pseudobaeospora lilacina KU528837

L . Pseudobaeospora wipapatiae KC464330

Pseudobaeospora pyrifera AY 176457

Callistosporium sp. AY 745702

1 Inocephalus sp. DQ457683

1 Leptonia canescens AF261307

Nolanea sericea DQ367423

Nolanea strictior AF042620

1 -— Rhodocybe mundula AY700182

Clitopilus prunulus AY700181

Neohygrophorus angelesianus DQ470814

- Dendrocollybia racemosa AF042598

Clitocybe dealbata AF223175

-— Lepista irina DQ234538

Ossicaulis lignatilis AF261396

~ Tricholomella constricta AF223188

Lyophyllum leucophaeatum AF223202

Calocybe carnea AF223178

Asterophora lycoperdoides AF223190

Termitomyces sp. DQ110875

Podabrella microcarpa AF042587

Lyophyllum decastes AF042583

Lyophyllum sp. DQ094785

Collybia tuberosa AY639884

Clitoeybe nebularis DQ457658

Clitocybe subditopoda AY691889

08'— Clitocybe candicans AY 645055

Tricholoma aestuans AY 700197

_- Tricholoma myomyces2 U76459

Tricholoma myomyces| AF518660

Tricholoma inamoenum AY293215

Leucopaxillus albissimus AF042592

Tricholoma saponaceum AY 647209

Tricholoma matsutake U62964

Catathelasm ventricosum DQ089012

1 Clitocybe subvelosa AY 647208

—__—— —— Cyathus striatus AF336247

Mycena amicta DQ457692

Mycena aff. pura DQ457688

Mycena galericulata AY 647216

0.861 Panellus stypticus AF518634

Mycena galericulata2 AF042636

om Mycena plumbea DQ470813

Agaricus bisporus AY 635775

os4 0.99

0.02

Fic. 2. Fifty percent majority-rule Bayesian cladogram of Pseudobaeospora and related taxa based

on nrLSU sequence analysis. Node support is indicated by Bayesian posterior probabilities >0.5 on

branches. Pseudobaeospora lilacina sequences are presented in boldface.

332 ... Wu &al.

Taxonomy

Pseudobaeospora lilacina X.D. Yu, Ming Zhang & S.Y. Wu, sp. nov. FIG. 3

MycoBank MB 815567

Differs from other species in the Frieslandica group by its lilac pileus, stubbly stipe, and

ellipsoid spores.

Type: China. Guangdong Province, Guangzhou City, Baiyun Mountain, on grass, 28 May

2013, Zhang M 3459 (Holotype, SYAU FUNGI-009; GenBank KU528840, KU528836).

EryMo.ocy: The epithet Jilacina refers to the lilac pileus color.

Piteus 1-3 cm diam, plano-convex to almost flat, dry; surface pale mauve,

colonial buff when old, slightly squamulose, darker at center, hygrophanous;

margin flexuous to involute. CONTEXT 0.1-0.2 cm thick at stipe, thin at

margin, whitish to tinged with pileus’ color. LAMELLAE adnate to uncinate,

arched, <3 mm broad, unequal, distant, with lamellulae of 2-4 lengths but not

anastomosing, pale mauve to mauve, undulate and thin at edge. StipE 2-3 x

0.2-0.5 cm, cylindrical, somewhat expanded at base, sometimes pliant, pale

Congo pink, fibrous, striate, central, pruinose to flocculose overall, bruising

indigo-bluish. OporR inconspicuous.

BASIDIOSPORES 2.5-3.5(-4.5) x 3-5(-6.5) um, Q = 1.38, broadly ellipsoid

to ellipsoid, hyaline, thick-walled, dextrinoid, minutely rough and with a

distinct hilar appendage under scanning electron microscope. BAstp1A 14-25 x

3.0-5.8 um, 4-spored and 2-spored. HyMENIUM hyphae 3.5-7 um diam, hyaline,

thin-walled, non-amyloid; cystidia absent. HYMENOPHORAL TRAMA 100-180

um wide, regular, hyphae 2.8-7.3 um, thin-walled. PILEIPELLIs a trichoderm of

ascending cylindrical hyphae, 6-10 um diam, thin-walled, hyaline; suprapellis

composed of regular parallel hyphae, 4.2-8.3 um diam, thin-walled, hyaline,

color unchanging in 5% KOH. CLAMP CONNECTIONS present in all parts of the

basidiocarp.

Hasirt saprophytic, solitary on naked soil or on grass.

ADDITIONAL SPECIMENS EXAMINED: CHINA. GUANGDONG PROVINCE, Zhaoqing

City, Fengkai County, Heishiding Nature Reserve, 3 June 2013, Yu XD 5546 (SYAU

FUNGI-011; GenBank KU528842, KU528838); Guangzhou City, Baiyun Mountain, 28

May 2013, Yu XD 3450 (SYAU FUNGI-010; GenBank KU528841, KU528837); 22 June

2008, Li TH, Chen XL, Li Y (GDGM25609; GenBank KX266951, KX266952); 22 May

2011, Xu J (GDGM44799); 25 May 2011, Xu J, Chen XL, Zhang M (GDGM28870); 27

May 2011, Xu J, Qiu CS, Zhang M (GDGM28919).

Discussion

Pseudobaeospora lilacina represents the first report of Pseudobaeospora

in China. Based on morphology, the new species should be placed in the

Pseudobaeospora lilacina sp. nov. (China) ... 333

EHT = 15.00 kV Signal A= SE2 Date :12 Dec 2015 ] EHT = 15.00 kV Signal A = SE2 Date :12 Dec 2015

WO= 8.0mm Mag= 30,.00KX Time 19:42:54 WD = 8.0mm Mag= 20.00KX Time :19:57:22

Fic. 3. Pseudobaeospora lilacina (holotype, SYAU FUNGI-009). A, B: Macroscopic habit.

C, D: Surface of spores. Scale bars: A, B = 1 cm; C = 1 um; D = 2 um.

Frieslandica group: colored basidiocarp, presence of clamp connections, and

non-hymenidermoid pileipellis (Bas 2003). However, its pale mauve pileus

and stubbly stipe easily distinguish P lilacina from the other Frieslandica

species. Although most of the other species have purple tones in the pileus,

their colors are conspicuously deeper (Bas 2003) than those found in

P. lilacina. Pseudobaeospora subglobispora Bas from Asia shares a similar

pileus color, but is distinguished from P lilacina by its shorter (0.4—1 cm) stipe

and obvious cheilocystidia. Three additional Asian species—P. citrina Rawla,

P. lavendulamellata Arnolds et al., P. oligophylla—are easily distinguished from

P lilacina: P. citrina by a greenish yellow pileus (Bas 2003), P lavendulamellata

by a dull violet pileus (Arnolds et al. 2004), and P oligophylla by a dark

violaceous pileus (Bas 2003). Furthermore, in P. oligophylla there are no clamp

connections in the basidiocarp hyphae (Bas 2003), and P lavendulamellata

has obvious cheilocystidia and its pileipellis discolors in 5% KOH (Arnolds

et al. 2004).

334 ... Wu &al.

We found only three Pseudobaeospora species—P. deckeri, P pyrifera,

P. wipapatiae—with available GenBank sequences, indicating the deficiency

of molecular research on the genus. However, our molecular analyses

provide some clues as to the taxonomic position of Pseudobaeospora. Both

ITS and nrLSU sequence analyses (Fics 1, 2) placed the Pseudobaeospora

species and Callistosporium graminicolor into one clade supported by strong

Posterior Probabilities (PP = 1), with the clade clustering with Lyophyllaceae

or Entolomataceae. Our result is inconsistent with the previous nrLSU

sequence analysis by Desjardin et al. (2014), which placed Pseudobaeospora

in Tricholomataceae. ‘The reason for the inconsistency is not clear but may

be explained by the low number of sequences available or taxa selected for

analysis. Even so, our result supports the observation by Vellinga (2004) that

Pseudobaeospora does not belong in Agaricaceae.

Pseudobaeospora will require more sampling and molecular analyses before

its taxonomic position can be established.

Acknowledgments

This study was supported by the National Natural Science Foundation of China

(No. 31200011) and Natural Science Foundation of Liaoning Province (201602668).

The authors are very grateful to Prof. Taihui Li (Guangdong Institute of Microbiology),

Prof. Alfredo Justo (Biology Department, Clark University), and Prof. Haisheng Yuan

(Institute of Applied Ecology, Chinese Academy of Sciences) for pre-submission review.

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MY COTAXON

April-June 2017— Volume 132, pp. 337-341

https://doi.org/10.5248/132.337

Gymnopilus dilepis, a new record in Thailand

NAKARIN SUWANNARACH *, JATURONG KUMLA ’,

KRIANGSAK SRI- NGERNYUANG * & SAISAMORN LUMYONG ‘*

' Department of Biology, Faculty of Science, Chiang Mai University,

Chiang Mai, 50200, Thailand

’ Faculty of Architecture and Environmental Design, Maejo University,

Chiang Mai, 50290, Thailand

* CORRESPONDENCE TO: saisamorn.l@cmu.ac.th

ABSTRACT—Specimens collected in Thailand have been identified as Gymnopilus dilepis,

based on morphology and ITS molecular analysis. A description and illustration are provided.

This is the first record of this fungus from Thailand.

Key worps—Agaricales, Cortinariaceae, gill fungus, taxonomy

Introduction

Karsten (1879) erected the genus Gymnopilus, with Agaricus liquiritiae

Pers. [= G. liquiritiae (Pers.) P. Karst.] as the type species. Gymnopilus has

been treated as a member of Cortinariaceae, Agaricales (Singer 1986). This

genus of saprotrophic fungi is distributed worldwide in temperate and tropical

regions, usually occurring on dead wood (Singer 1986; Guzman-Davalos

2003), and approximately 200 species have been reported (Kirk et al. 2008).

Four Gymnopilus species—G. aeruginosus (Peck) Singer, G. junonius (Fr.) P.D.

Orton, G. penetrans (Fr.) Murrill, and G. punctifolius (Peck) Singer—have

been reported from Thailand (Chandrasrikul et al. 2011). During a taxonomic

survey of macrofungi collected in northern Thailand, we collected specimens

that corresponded to the description of G. dilepis, a species previously reported

from Australia, India, Indonesia, Japan, Malaysia, Papua New Guinea, Sri

Lanka, and the United Kingdom (Pegler 1986, Treu 1998, Guzman-Davalos

338 ... Suwannarach & al.

2003, Guzman-Davalos et al. 2003, Thomas et al. 2003, Rees et al. 2004, Kasuya

et al. 2016). Here we describe and illustrate the morphological characters of the

Thai material and provide results from ITS sequence analysis.

Materials & methods

Morphology studies

Basidiocarps were collected from Lampang Province, Thailand, in 2015 and wrapped

in aluminum foil or kept in plastic specimen boxes for transport to the laboratory.

Notes on macromorphological features and photographs were obtained within 24 h of

collection. Color names and codes follow Kornerup & Wanscher (1978). The specimens

were dried at 40—45 °C; sections of dried material were mounted in 95% ethanol

and rehydrated in distilled water, 3% KOH, or Melzer’s reagent for microscopical

examination; at least 50 measurements were made of each structure. The collections are

deposited in the herbarium of the Research Laboratory for Excellence in Sustainable

Development of Biological Resources, Faculty of Science, Chiang Mai University,

Thailand (SDBR-CMU).

Molecular studies

Genomic DNA of two specimens was extracted from fresh tissue using a Genomic

DNA Extraction Mini Kit (Favorgen, Taiwan). The internal transcribed spacer (ITS)

region of the ribosomal RNA gene was amplified by the polymerase chain reaction

(PCR) with ITS4 and ITS5 primers under the following thermal conditions: 94 °C

for 2 min; 35 cycles of 95 °C for 30 s, 50 °C for 30 s, 72 °C for 1 min; and 72 °C for

10 min. PCR products were checked on 1% agarose gels stained with ethidium bromide

under UV light and purified using NucleoSpin’ Gel and a PCR Clean-up Kit (Macherey-

Nagel, Germany), following the manufacturer's protocol. The purified PCR products

were directly sequenced. Sanger sequencing was carried out by 1°’ Base Company

(Kembangan, Malaysia) using the ITS4 and ITS5 primers. Sequences were used to query

the GenBank database via BLAST (http://blast.ddbj.nig.ac.jp/top-e.html).

Results

Taxonomic description

Gymnopilus dilepis (Berk. & Broome) Singer, Lilloa 22: 560 (1951). Fig. 1

Pileus 20-50 mm in diameter, convex to plano-convex, surface light orange

(6A5) towards the center, pale orange (5A3) to light orange (5A4) towards the

margin, squamulose; squamules ruby (12D8) to violet brown (10F8), erect and

comparatively denser at the center, sparse and appressed in the middle, sparse

or almost absent towards the margin; margin decurved to plane. Context

concolorous with the pileus surface. Lamellae adnate to subdecurrent, light

orange (5A5) to orange (5A6). Stipe 25-60 x 3—9 mm, central, almost equal

or slightly tapering basally, surface pale orange (5A3), becoming dark brown

Gymnopilus dilepis, new for Thailand ... 339

Fic. 1. Gymnopilus dilepis (SDBR-CMU-NK0O116).

A: basidiocarps; B: basidiospores; C: basidia and basidiospores; D: cheilocystidia; E: caulocystidia.

Scale bars: A = 10 mm, B—D = 5 um, E= 10 um.

(6F6) towards base. Veil present. Odor slight. Pileus and stipe surface darkened

to brownish orange (6C8) with aqueous KOH. Spore print light brown (6D8).

Basidiospores 6—7.5 x 4.8—6 um, Q = 1.23-1.5, broadly elliptical to ellipsoid,

with obtuse apex, slightly thickened walls, verrucose, warts medium to large,

dextrinoid, orange-brown in KOH. Basidia 20—27.5 x 5—8.5 um, clavate to

cylindric-clavate, hyaline, 4-spored, with basal clamp connection, sterigmata

<5.5 um long. Pleurocystidia absent. Pseudocystidia 20—27 x 7—8.5 um, clavate-

rostrate or subfusiform, thin-walled, with granulose to homogeneous orange-

brown contents. Cheilocystidia 20—30.2 x 8-12 um, apex 2.2—4.5 um diam.,

340 ... Suwannarach & al.

utriform, ranging from clavate with a wide rostrum to lageniform with a short

neck and a non-capitate or subcapitate apex, thin-walled, with basal clamp

connection, hyaline to pale yellow. Hymenophoral trama subregular, hyphae

2-20 um diam., thin-walled, hyaline to pale yellow. Subhymenium inflated-

ramose. Pileal trama radial, hyphae 2—20.5 um diam., thin-walled, hyaline to

pale yellow. Pileipellis a cutis, hyphae 3-15 um diam., thin-walled, coarsely

encrusted with brown pigment. Stipe trama composed of hyphae 2—25 um

diam., parallel, thin-walled, pale yellow. Stipitipellis a cutis, hyphae 2-12 um

diam., thin-walled with pale yellow to brown wall pigment. Caulocystidia

18.2—65 X 5.5-14.5 um, cylindrical, clavate, narrowly utriform, with obtuse or

subcapitate apex, hyaline, some with granulose, pale yellow or orange-brown

content, thin-walled, in tufts at the stipe apex. Clamp connections present on

all hyphae.

SPECIMENS EXAMINED—THAILAND, LAMPANG PROVINCE, Mae Moh District,

18°40’52”N 98°52’10”E, elevation 550 m, on dead wood in a deciduous forest, 8

November 2015, Suwannarach N & Lumyong S$ (SDBR-CMU-NKO116; GenBank

KX639496); Kumla J & Suwannarach N (SDBR-CMU-JK0142; GenBank KX639497).

Molecular analysis

The ITS sequences of specimens SDBR-CMU-NKO116 (644 bp) and

SDBR-CMU-JK0142 (638 bp) were deposited in GenBank. All ITS sequences

obtained in this study showed 100% similarity with G. dilepis KT368680 and

KT368682, from Japan.

Discussion

Both specimens collected in northern Thailand were initially identified

as G. dilepis based on descriptions by Guzman- Davalos (2003) and Thomas

et al. (2003). This species is easily distinguished from G. penetrans, which

produces a gray-brown to dark brown pileus with a white to grayish-white

tomentose-arachnoid velum covering the whole surface (Holec 2005). The

smaller basidiospores (4.0—6.0 x 3.5—5.0 um) of G. punctifolius and the longer

basidiospores (8.0—10.5 x 5.5—7.2 um) of G. junonius clearly distinguish these

two species from G. dilepis (Singer 1951; Holec 2005).

Gymnopilus dilepis is closely related to G. purpuratus (Cooke & Massee)

Singer and G. norfolkensis B.J. Rees & Lepp, from which it is distinguished

by differences in spore size: basidiospores of G. purpuratus differs are longer

(7.5-8.7 X 4.8-5.7 um; Rees et al. 2004) while those of G. norfolkensis are

narrower (6.4—7.2 xX 4.0-5.2 um; Rees and Lepp 2000). Gymnopilus dilepis

also resembles G. lepidotus Hesler, but the two species differ in the form of

Gymnopilus dilepis, new for Thailand ... 341

the cheilocystidia; additionally G. lepidotus is known only from Mexico and

the United States (Hesler 1969, Guzman-Davalos 2003, Guzman-Davalos et al.

2003).

Our molecular analysis also confirmed the two Thai specimens as

representing G. dilepis. The combination of morphological and molecular

characters confirms G. dilepis as a new record for Thailand.

Acknowledgements

This work was supported by grants from Chiang Mai University and the Thailand

Research Fund, Research-Team Association Grant (RTA5880006). We are grateful

Dr. Eric H.C. McKenzie and Dr. Steven L. Stephenson for their helpful comments and

careful review of this article.

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Guzman- Davalos L. 2003. Type studies of Gymnopilus (Agaricales) I. Mycotaxon 86: 395-423.

Guzman-Davalos L, Mueller GM, Cifuentes J, Miller AN, Santerre A. 2003. Traditional infrageneric

classification of Gymnopilus is not supported by ribosomal DNA sequence data. Mycologia 95:

1204-1214.

Hesler LR. 1969. North American species of Gymnopilus. Mycologia Memoirs 3. 117 p.

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collection from other European countries. Acta Musei Nationalis Pragae, Series B, Historia

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Kasuya T, Kobayashi E, Kurokawa H, Pham HND, Hosaka K, Terashima Y. 2016. Three

species of Gymnopilus newly recorded in Japan. Japanese Journal of Mycology 57: 31-45.

http://doi.org/10.18962/jjom.57.1_31

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

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Rees BJ, Lepp H. 2000. A new species of Gymnopilus from Norfolk Island. Australasian Mycologist

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purple-coloured Gymnopilus species in Europe. Australasian Mycologist 22: 57-72.

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Thomas AT, Guzman- Davalos L, Manimohan P. 2003. A new species and new records of Gymnopilus

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MY COTAXON

April-June 2017— Volume 132, pp. 343-356

https://doi.org/10.5248/132.343

A reassessment of Hourangia cheoi from Yunnan, China

MATTEO GELARDI ', ALFREDO VIZZINI” & ENRICO ERCOLE ”

'Via Angelo Custode 4A, I-00061 Anguillara Sabazia, RM, Italy

? Department of Life Science and Systems Biology,

University of Turin, Viale PA. Mattioli 25, I-10125 Torino, Italy

" CORRESPONDENCE TO: alfredo. vizzini@unito.it

ABSTRACT—A recent collection of Hourangia cheoi (Boletaceae, Boletales) from near the

type locality in Yunnan Province, southwestern China, is described in detail and illustrated,

and its nrITS and nrLSU rDNA sequences are included in molecular phylogenetic analyses.

Comparisons with closely related xerocomoid taxa are also provided.

Key worps—biogeography, ectomycorrhizal fungi, taxonomy, Xerocomus

Introduction

During repeated mycological field expeditions carried out by the first

author during 2011-13 in central, southwestern, and southeastern China

(Beijing, Shaanxi, Yunnan, and Guangdong Provinces), several new or

noteworthy fungal species were recorded and documented (Gelardi et al.

2012, 2013, 2014, 2015a,b; Vizzini et al. 2012, 2014; Battistin et al. 2014,

Gelardi 2014, Picillo & Gelardi 2014). Amongst the several bolete species

found in the surveyed areas, a single specimen with a xerocomoid habit was

collected near Dali, Yunnan, and later identified from morphological and

molecular analyses as Hourangia cheoi.

Chiu (1948) originally described the species in the genus Boletus Fr. In the

same paper the Chinese author described Boletus punctilifer; both species were

subsequently illustrated (Chiu 1957) and later combined in Xerocomus Quel.

(Tai 1979). However, Zhu et al. (2015) reduced B. punctilifer to synonymy

under B. cheoi after examining numerous specimens, including the holotypes

of both names.

344 ... Gelardi, Vizzini & Ercole

As a result of intensive research on Chinese xerocomoid boletes, Zhu et al.

(2015) erected a new genus Hourangia based on Boletus cheoi, including three

other species originally described as B. microcarpus Corner, B. nigropunctatus

W.E. Chiu, and Phylloporus pumilus M.A. Neves & Halling. Hourangia is related

to Xerocomus s.str. and sister to Phylloporus Quél. (Wu et al. 2014, Zhu et al.

2015, Henkel et al. 2016), and despite the phylogenetic and morphological

affinities with these genera, it is readily distinguished by a finely squamulose

pileus surface, tubes that are three to seven times longer than the thickness of

pileus context, whitish basal mycelium, context turning blue then reddish and

finally fading to blackish-brown on exposure, a bacillate spore ornamentation

(visible only with a scanning electron microscope), a “Phylloporus-type”

hymenophoral trama, ectomycorrhizal association with Fagaceae, Pinaceae,

and Dipterocarpaceae, and its eastern and southeastern Asian distribution

(Zhu et al. 2015; pers. obs.).

The aim of the present contribution is to provide new insights and

iconography on Hourangia cheoi.

Materials & methods

Collection site and sampling

The specimen was collected in the surroundings of Dali, Yunnan Province, China,

and is deposited in ZT (Thiers 2017) and in the personal herbarium of Matteo

Gelardi (MG). Author citations follow Index Fungorum (www.indexfungorum.org/

authorsoffungalnames.htm).

Morphological studies

Macroscopic descriptions, habitat notations, and associated plant communities

were based upon detailed field notes from fresh basidiomes. Color terms in capital

letters (e.g. Myrtle Green, pl. VIII) are from Ridgway (1912). The specimen was

photographed in habitat using a Nikon D3100 camera. Micro—morphologic features

were observed from dried material; sections were rehydrated in water, 5% potassium

hydroxide (KOH), or ammoniacal Congo Red. Structures and anatomical features

were observed and measured in Congo Red; color and pigments were observed in

KOH. Measurements were determined at 1000x with a calibrated ocular micrometer

(Nikon Eclipse E200 optical light microscope). Basidiospores were measured from

the mature hymenophore; dimensions are given as (minimum -) average + standard

deviation (-maximum), Q = length/width (L/W) ratio with the extreme values in

parentheses, Qm = average quotient (L/W ratio) + standard deviation, and average

spore volume was approximated as a rotation ellipsoid (V = (1.L.W’) /6 + standard

deviation). The notation [n/m/p] indicates that measurements were made on “n”

randomly selected basidiospores from “m” basidiomes of “p” collections. The width

of each basidium was measured at the widest part, and the length was measured from

Hourangia cheoi reassessed (China) ... 345

the apex (sterigmata excluded) to the basal septum. Metachromatic, cyanophilic, and

iodine reactions were tested by staining the basidiospores in Brilliant Cresyl blue,

Cotton blue and Melzer’s reagent, respectively. Line drawings of microstructures were

made freehand based on photomicrographs of rehydrated material.

DNA extraction, PCR amplification and sequencing

Total DNA was extracted from the dry specimen (MG484) with the DNeasy

Plant Mini Kit (QIAGEN, Milan, Italy) according to the manufacturer’s instructions.

PCR primers ITSIF and ITS4 (White et al. 1990, Gardes & Bruns 1993) for the

nrITS region, LROR, LR1, LR5 and LR7 (Vilgalys & Hester 1990, Cubeta et al. 1991,

Van Tuinen et al. 1998) for the nrLSU region, were employed for PCR amplification

and sequencing purposes. The PCR protocol comprised a hot start at 95°C (5 min),

followed by 35 cycles of 94°C (45 s), 54°C (30 s), and 72°C (45 s), and a final 72°C

step (10 min). PCR products were checked in 1% agarose gels before purification and

sequencing. Chromatograms were searched for putative sequencing reading errors.

The sequences are deposited in GenBank and their accession numbers are included

in Fires 1, 2.

Sequence alignment, dataset assembly and phylogenetic analyses

The sequences obtained in this study were checked and assembled using Geneious

v. 5.3 (Drummond et al. 2010) and compared to those available in GenBank using

the Blastn algorithm. Based on the BLASTn results (with sequences selected based

on greatest similarity) and the phylogenetic analysis by Zhu et al. (2015), sequences

were retrieved from GenBank and UNITE (http://unite.ut.ee/) databases for the

comparative phylogenetic analysis. Alignments were generated for each single LSU

and ITS dataset using MAFFT (Katoh et al. 2002) with default conditions for gap

openings and gap extension penalties. The two alignments were then imported into

MEGA v. 6.0 (Tamura et al. 2013) for manual adjustment. The best-fit models were

estimated by the Bayesian Information Criterion (BIC) using jModelTest v. 2.1.7

(Darriba et al. 2012) to provide a substitution model for each single alignment.

GTR+G models were chosen for both the ITS and LSU alignments.

Phylogenetic analyses were performed using the Bayesian Inference (BI) and

Maximum likelihood (ML) approaches. Phylloporus rhodoxanthus (JQ003654),

Xerocomus ferrugineus (JQ003657) and X. subtomentosus (KC215201) were chosen

as the outgroup taxa in the ITS analysis (Fic. 1); and Phylloporus imbricatus

(KF112398), P luxiensis (KF112490), P. pelletieri (AF456818), P. rubrosquamosus

(KF112391), “Xerocomus perplexus” (JQ003702), and X. subtomentosus (KC215222)

in the LSU analysis (Fie. 2), following Zhu et al. (2015). Bayesian and ML inferences

were performed online using the CIPRES Science Gateway website (Miller et al.

2010) and both methods were implemented as single software usage. BI phylogeny

using Monte Carlo Markov Chains (MCMC) was carried out with MrBayes v. 3.2.2

(Ronquist et al. 2012). Four incrementally heated simultaneous MCMC were run

over 10 M generations. Trees were sampled every 1000 generations resulting in an

overall sampling of 10,001 trees. The first 2500 trees (25%) were discarded as burn-

346 ... Gelardi, Vizzini & Ercole

in. For the remaining trees, a majority rule consensus tree showing all compatible

partitions was computed to obtain estimates for Bayesian Posterior Probabilities

(BPP). Branch lengths were estimated as mean values over the sampled trees.

ML estimation was performed through RAxML v. 7.0.4 (Stamatakis 2006) with

1000 bootstrap replicates (Felsenstein 1985) using the GTRGAMMA algorithm

to perform a tree inference and search for a good topology. Support values from

bootstrapping runs (MLB) were mapped on the globally best tree using the

‘fa option of RAxML and ‘-x 12345’ as a random seed to invoke the novel rapid

bootstrapping algorithm. Only BPP values >0.75 and/or MLB values >50% are

reported in the resulting trees.

Results

Both Bayesian and Maximum Likelihood analyses produced the same

topology; only the BI trees with both BPP and MLB values are shown (Fics 1, 2).

Hourangia nigropunctata KP136989

1/96 | - Hourangia nigropunctata KP 136988

Hourangia nigropunctata KP136991

O.98/77

Hourangia nigropunctata KP136992

1/00 4a|' Hourangia nigropunctata KP 136993

Hourangia nigropunctata KP 136990

Hourangia cheoi KP136994

Leo range cheoi KP137001

Hourangia cheoi KP136997

Hourangia cheoi KP136998

HOUrANGIA siroo Hourangia cheoi KP137000

Hourangia cheoi KP136999

Hourangia cheoi KP137002

Hourangia cheoi KP136996

Hourangia cheoi KP136995

Hourangia cheoi MG484 KX911873

Hourangia pumila JQ003627

4100

Hourangia sp. HKAS 68178 KP136987

Hourangia microcarpa KP136986

Xerocomus ferrugineus JQ003657

Phylloporus rhodoxanthus JQ003654

Xerocomus subtomentosus KC215201

0.05 expected changes per sites

Fic. 1. Bayesian phylogenetic analysis based on the ITS sequences of Hourangia species and

related taxa. Bayesian posterior probabilities (BPP; in bold) 20.75 and Maximum Likelihood

bootstraps (MLB) =50% are shown on the branches. Thickened branches indicate BPP =0.95

and MLB =70%. The GenBank/UNITE number is given for each sequenced taxon. The new

H. cheoi sequence is in bold.

Hourangia cheoi reassessed (China) ... 347

The ITS data matrix comprised a total of 22 sequences (including 21 from

GenBank); the alignment comprised 1160 characters and contained 526

(45.3%) variable sites, of which 284 (24.5%) were parsimony informative.

The LSU matrix consisted of 34 sequences (including 33 from GenBank);

the alignment comprised 830 characters and contained 144 (17.3%) variable

sites, of which 110 (13.2%) were parsimony informative.

Both in the ITS and LSU analyses, our collection and the Hourangia

cheoi sequences retrieved from GenBank form a strongly supported clade

(BPP = 1 and MLB = 100 in the ITS analysis; BPP = 0.95 and MLB = 95 in

the LSU analysis).

oases | Hourangia cheoi MG484 KX911874

Hourangia cheoj KP136948

Hourangia cheoi KP136952

Hourangia cheoi KP136953

Hourangia cheoi KP136950

Hourangia cheol KP136957

Hourangia cheoi KP136954

Hourangia cheoi KP136951

Hourangia cheoi KP136955

Hourangia cheoi KP136956

Hourangia cheoj KP136949

Hourangia cheoj KP136947

Hourangia cheoi KP136946

Hourangia cheoi KF112385 [epitype]

{ Hourangia nigropunctata KP136960

Hourangia nigropunctata KP136961

ta nigropunctata KP136965

Hourangia nigropunctata KP136959

Houran g ia 489 186 [| Hourangia nigropunctata kP136962

| Hourangia nigropunctata KP136963

| Hourangia nigropunctata KP136958

Hourangia nigropunctata KP136964

— Hourangia nigropunctata KF 112388

0.99/74 Rea 178 + Hourangia pumila JQ003681

Hourangia pumila Ja003682

Hourangia sp. HKAS 68178 KF112453

Hourangia microcarpa KP136945

Hourangia microcarpa KF112452

0.95/95

0.99/89 |

0.8/80

0.85/65

0.86/51

34057 ge Phylioporus imbricatus KF 112398

Phyiloporus luxiensis KF112490

—_ Phyloporus pelietieri AF456818

Phylloporus rubrosquamosus KF 112391

Xerocomus subtomentosus KC215222

‘Xerocomus perplexus” JQ003702

0,02 expected changes per sites

Fic. 2. Bayesian phylogenetic analysis based on the LSU sequences of Hourangia species and

related taxa. Bayesian posterior probabilities (BPP; in bold) 20.75 and Maximum Likelihood

bootstraps (MLB) =50% are shown on the branches. Thickened branches indicate BPP =0.95

and MLB =70%. The GenBank number is given for each sequenced taxon. The new H. cheoi

sequence is in bold.

348 ... Gelardi, Vizzini & Ercole

Taxonomy

Hourangia cheoi (W.F. Chiu) Xue T. Zhu & Zhu L. Yang,

Mycol. Progr. 14: 37 [4 of 10] (2015). FIGs 3, 4

MycoBank MB 810696

= Boletus cheoi W.F. Chiu, Mycologia 40(2): 215 (1948).

= Xerocomus cheoi (W.E. Chiu) EL. Tai, Syll. Fung. Sinicorum: 813 (1979).

= Boletus punctilifer W.F. Chiu, Mycologia 40(2): 216 (1948).

= Xerocomus punctilifer (W.F. Chiu) EL. Tai, Syll. Fung. Sinicorum: 814 (1979).

BASIDIOMES medium-small. PiLEus 4.3 cm diam., convex-pulvinate,

regularly shaped, moderately fleshy, soft; margin straight to faintly wavy,

curved downwards, not or only slightly extending beyond the tubes; surface

matte, dry, very finely squamulose, not cracked; cuticle evenly dark ochraceous

(Buckthorn Brown, pl. XV; Sayal Brown, pl. XXIX); slowly darkening to

dull brownish-black (Warm Sepia, pl. XXIX) on handling or when injured,

especially at the edge of squamules; subcuticular layer whitish (White,

pl. LIII). TuBes somewhat broad and decidedly longer than the thickness of

pileus context (<1.0 cm long), adnate, olive yellow (Javel Green, pl. V), turning

blue (Myrtle Green, pl. VIII) when cut. Pores forming a slightly convex or

ascendant surface, simple, not radially arranged, prominently wide and angular

(<1 mm in diam.), concolorous with the tubes and turning blue (Myrtle Green,

pl. VII) on bruising or when injured and finally fading to brownish (Snuff

Brown, pl. XXIX), without innate rusty brown stains at the orifice. STIPE 5.3 x

0.6 cm, slightly longer than pileus diameter, central, solid, firm, dry, faintly

curved, cylindrical but slightly swollen at the base, ending with a short taproot

at the very base; surface finely longitudinally fibrillose, devoid of reticulum

or any other kind of ornamentation, veils absent; beige-ochraceous (Cream

Buff, pl. XXX) throughout but paler (Cartridge Buff, pl. XXX) towards the

base, unchangeable when pressed; basal mycelium whitish (White, pl. LIID),

rhizomorphs brownish (Dresden Brown, pl. XV). CONTEXT soft textured and

thin in the pileus (<0.4 cm thick in the central zone), a little more fibrous

in the stipe, whitish (White, pl. LIII) throughout but pale yellow (Martius

Yellow, pl. IV) above the hymenophore; turning light blue (Pale Methyl Blue;

Light Cerulean Blue, pl. VIII; Sky Blue, pl. XX) in the pileus and stipe/pileus

connecting zone when exposed to air but changing within twenty seconds to

bright flesh pink (Buff Pink, Congo Pink, pl. XXVIII) and eventually fading to

grayish (Light Mouse Gray, pl. LI) in patches, elsewhere in the stipe directly

turning to pinkish-red (Congo Pink, Japan Rose, Terra Cotta, pl. XXVIII)

after cutting, especially towards the base, then fading to dirty grayish (Mouse

Hourangia cheoi reassessed (China) ... 349

Fic. 3. Hourangia cheoi (MG484).

Basidiome in habitat. Scale bar = 2 cm. Photo by M. Gelardi.

Gray, pl. LI) or grayish—-black (Blackish Mouse Gray, Sooty Black, pl. LI); sub-

hymenophoral layer yellowish (Martius Yellow, pl. IV); exsiccate brownish

(Dresden Brown, pl. XV). SMELL hardly perceptible, fungoid. Taste mild.

SPORE PRINT not obtained.

Basip1osPorss [40/1/1] (9.8—)11.2 + 0.8(-13.1) x (4.0-)4.6 + 0.2(-5.0) um,

Q = (2.17-)2.22-2.72(-2.75), Qm = 2.44 + 0.14, V = 126 + 20 um’, inequilateral,

ellipsoid-fusiform to fusiform in side view, ellipsoid-fusiform in face view,

smooth under the light microscope, with a prominent apiculus and shallow

suprahilar depression, apex rounded, moderately thick-walled (0.5-0.7 um),

straw yellow to bright yellow colored in KOH, having one or (less frequently)

two large oil droplets when mature, rarely pluri-guttulate, inamyloid to faintly

dextrinoid, cyanophilic and with an orthochromatic reaction; no anomalous

spores observed. Bastp1A 28-37(-40) x 9-11 um (n = 10), cylindrical-clavate

to clavate, moderately thick-walled (0.5-0.8 um), predominantly 4-spored but

350 ... Gelardi, Vizzini & Ercole

also 2- or (sporadically) 3-spored, usually bearing relatively long sterigmata

(2-7 um), hyaline to very pale yellowish and containing straw-yellow oil

guttules in KOH, bright yellow to orange-reddish (barely dextrinoid) in

Melzer'’s, without basal clamps; basidioles subcylindrical to faintly clavate,

similar in size to basidia. CHEILOCySTIDIA (40-)51-76(-82) x 6-11 um

(n = 17), very common, moderately slender, projecting, straight to sometimes

flexuous, cylindrical to cylindrical-fusiform, rarely subclavate and occasionally

subcapitate or mucronate, with rounded tip, smooth, moderately thick-walled

(0.8-1.0 um), hyaline or with a very pale yellowish pigment in KOH, bright

yellow (inamyloid) in Melzer’s and with an orthochromatic reaction, without

epiparietal encrustations. PLEUROCYSTIDIA (62-)67-110(-113) x 7-23 um

(n=21), decidedly frequent, colorand chemical reactions similar to cheilocystidia

but slightly different in shape and distinctly longer, straight, slender, cylindrical-

fusiform or fusiform to ventricose-fusiform, less frequently lageniform,

sometimes with a narrow and long neck, apex rounded. PsEUDOCYSTIDIA not

recorded. PILEIPELLIS a trichoderm or palisadoderm consisting of subparallel

to loosely interwoven, elongated, cylindrical, rarely branched hyphae tending

to remain erect in the outermost layer and not embedded in gelatinous matter;

terminal elements usually long, slender and markedly swollen (cylindrocytes),

cylindrical to sausage-shaped, less frequently cystidioid, subclavate or peanut-

shaped, acorn-shaped or bullet-shaped, apexrounded-obtuse, (36-)41-88(-103)

x (12-)15-29(-33) um (n = 26), intermixed with scattered longer, filamentous

and sinuous cells (up to 127 x 13 um), moderately thick-walled (<1 um);

pileipellis elements often appear dissociated and easily separable from one

another, hyaline to (more often) yellowish in KOH, bright yellow to orange-

reddish (weakly dextrinoid) in Melzer’s, smooth; subterminal elements similar

in shape, size, color and chemical reactions to terminal elements. STIPITIPELLIS

a texture of slender, subparallel to parallel and longitudinally running, smooth

walled, adpressed hyphae, 4-19 um wide, hyaline in KOH; the stipe apex

covered by tufts of caulohymenium consisting of sterile caulobasidioles, very

sparse 2-/3-spored fertile caulobasidia similar in shape, size and color to

hymenial basidia; CauLocystTip1A abundant, long, subcylindrical to fusiform,

ventricose-fusiform or lageniform, projecting, (33-)44-82(-92) x 7-10(-15) um

(n = 14), apex rounded, wall <1 um thick, hyaline to yellowish in KOH.

LATERAL STIPE STRATUM under the caulohymenium absent. STIPE TRAMA

hyphae longitudinal, densely arranged, subparallel to moderately interwoven,

filamentous, smooth to rarely encrusted by a very subtle granular, brownish

pigment, inamyloid to barely dextrinoid, 4-20 um diam. HYMENOPHORAL

Hourangia cheoi reassessed (China) ... 351

d| |

WaT

V {| va)

ih |

ae |

SAE m7,

S Ni | lam

SS aN

—_

SSA || \\

Fic. 4. Hourangia cheoi (MG484). a. Basidiospores. b. Basidia. c. Cheilocystidia. d. Pleurocystidia.

e. Hymenophoral trama. f. Pileipellis. Scale bars: a, b = 10 um; c-e = 20 um; f = 40 um.

Drawings by M. Gelardi.

TRAMA bilateral divergent (“Phylloporus-type’), hyphae very slightly divergent

to nearly subparallel and tightly arranged, non-gelatinous [lateral strata hyphae

in transversal section constricted at septa and distinctly broader than the

mediostratum hyphae, touching or almost touching each other, (0-)2-4 um

apart, (7—-)10-21(-24) um diam., relatively thick-walled (<1 um)], hyaline in

KOH, inamyloid in Melzer’s; lateral strata (15-)20-40 um thick, mediostratum

10-40 um thick, consisting of a tightly adpressed, non-gelatinous bundle of

hyphae, 4-12(-15) um diam.; in Congo Red the mediostratum is concolorous

with the lateral strata. RHIZOMORPHS consisting of parallel and densely

arranged, unbranched, cylindrical, moderately thick-walled (<1 um), smooth

hyphae, 5-10(-40) um diam., yellowish-orange to reddish-brown in KOH,

inamyloid to weakly dextrinoid in Melzer’s. CLAMP CONNECTIONS absent in all

tissues. HYPHAL SYSTEM monomitic. ONTOGENY gymnocarpic.

352 ... Gelardi, Vizzini & Ercole

ECOLOGY & DISTRIBUTION—Solitary or gregarious on soil in mixed forests

dominated by Fagaceae (Quercus, Castanopsis, Lithocarpus) and Pinaceae

(Keteleeria, Pinus); reported from southwestern China and Japan; distribution

limits unknown.

SPECIMEN EXAMINED: CHINA, YUNNAN PROVINCE, Xiaguan County, Cangshan

Mountain National Geopark, Gantong Temple, 25°39’03”N 100°09’58”E, 2260 m a.s.l.,

on a slope facing east in a subtropical montane forest hosting a country graveyard, a

single mature specimen with Pinus yunnanensis Franch. and P. armandii Franch. with

the presence of Quercus variabilis Blume, 19 Sep 2012, E. Horak & M. Gelardi (E. Horak

13448 (herb. ZT), MG484; GenBank KX911873, KX911874).

ASSOCIATED FUNGI: Other fungal species found in the same habitat are Amanita

parvipantherina Zhu L. Yang et al. and Cantharellus appalachiensis R.H. Petersen.

Discussion

The combination of morphological and molecular analyses confirmed

our identification of Hourangia cheoi. This eye-catching species appears to

be well outlined, especially due to the recent re-description provided by

Zhu et al. (2015). Although somewhat variable in its appearance, H. cheoi

is easily recognized in the field based on the following set of macroscopic

characters: its 1) small to medium stature, 2) finely squamulose, ochraceous

to brownish or reddish-brown pileus surface that slowly darkens when

handled, 3) hymenophore that quickly blues when injured, 4) <2 mm diam.

angular pores, 5) brownish to pale ochraceous fibrillose stipe that grades into

beige towards the base, and 5) whitish context that in the pileus turns light

blue, then pinkish-red, and finally grayish or dirty brown but in the stipe

discolors directly reddish then brownish or grayish-black upon exposure.

Diagnostic anatomical features include the 1) medium-sized boletoid

basidiospores with a bacillate surface under SEM, 2) very large ($113 x 23 um)

prominent pleurocystidia, and 3) trichodermal pileipellis comprising swollen

cylindrical to sausage-shaped terminal cells up to 33 um diam. (Chiu 1948,

Zhu et al. 2015; pers. obs.).

Several accounts have treated extensively or cited Hourangia cheoi under

the specific epithets “cheoi” or “punctilifer” (Zang 1986, 1996, 2006; Li &

Song 2000, 2003; Wang 2004, Zhang et al. 2012, Zang et al. 2013), and it has

generally been regarded as edible (Li & Song 2002, Dai et al. 2010), albeit

in most cases the real identity of the mushroom in cited reports cannot be

established with certainty.

The type species of Xerocomus, X. subtomentosus (L.) Queél., differs from

Hourangia cheoi by its finely tomentose pileus, pale yellowish context with

Hourangia cheoi reassessed (China) ... 353

pinkish hues in the lower third of the stipe and which turns light blue in the pileus

and pileus/stipe connection zone but without an eventual reddish to brownish

discoloration, stipe usually coarsely ribbed or roughly pseudoreticulate,

trichodermal pileipellis consisting of narrow filamentous hyphae (terminal

elements averaging 40 x 12 um), obviously smaller pleurocystidia, temperate

distribution, and obligate ectomycorrhizal association with broadleaved trees

(mainly Fagaceae and Corylaceae) (Alessio 1985, Breitenbach & Kranzlin 1991,

Engel et al. 1996, Lannoy & Estadés 2001, Ladurner & Simonini 2003, Watling

& Hills 2005, Taylor et al. 2006, Klofac 2007, Sutara et al. 2009, Knudsen &

Taylor 2012, Galli 2013).

For morphological comparisons of H. cheoi with the other three Hourangia

species, consult the dichotomous key in Zhu et al. (2015).

Acknowledgments

Special thanks to Prof. Zhu L. Yang of the Key Laboratory for Plant Diversity and

Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences,

for inviting and supporting logistic help during Gelardi’s stay in Yunnan Province

and for fieldwork opportunities. The first author is also indebted to G. Wu (Kunming,

China) and other graduate and undergraduate students for their assistance in collecting

fungi in the field. I. Krisai-Greilhuber (University of Vienna, Austria) and G. Simonini

(Reggio Emilia, Italy) are gratefully credited for reviewing the manuscript and providing

suggestions and comments. The first author's gratitude is also extended to E. Horak and

his wife (Innsbruck, Austria) for their companionship during field investigation and for

sharing their invaluable life-long experience on mycological topics.

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MY COTAXON

April-June 2017— Volume 132, pp. 357-360

https://doi.org/10.5248/132.357

Bharatheeya coronata sp. nov., a conidial fungus from Brazil

Lucas BARBOSA CONCEICAO ', MARCOS FABIO OLIVEIRA MARQUES ’,

JOSIANE SANTANA MONTEIRO *, Luis FERNANDO PASCHOLATI GUSMAO °°,

FLAVIA RODRIGUES BARBOSA 3? & RAFAEL F. CASTANEDA-RUIZ *?

1 Universidade Estadual de Feira de Santana,

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

? Universidade do Estado da Bahia, Departamento de Educacdo,

Campus VII, BR407 Km 127, 48970-000, Senhor do Bonfim, Bahia, Brazil

3 Universidade Federal do Mato Grosso, Instituto de Ciéncias Naturais, Humanas e Sociais,

Avenida Alexandre Ferronato, 1200, 78557-267, Sinop, Mato Grosso, Brazil

‘Instituto de Investigaciones Fundamentales en Agricultura (INIFAT) Alejandro de Humboldt,

Calle 1 Esq. 2, C.P. 17200, Santiago de Las Vegas, C. Habana, Cuba

* CORRESPONDENCE TO: “lgusmao@uefs.br; * rfcastanedaruiz@gmail.com

Asstract—Bharatheeya coronata, collected from decaying twigs in Bahia state, Brazil, is

described and illustrated. The new species differs from both other Bharatheeya species by

its smooth conidiophores, terminal conidiogenous cells that are truncate at the apex, and

coronate, 1-distoseptate, pale brown conidia.

KEY WORDS—asexual ascomycete, hyphomycetes, taxonomy, tropical fungi

Introduction

The diversity of saprobic conidial fungi in tropical regions is still largely

unknown, although the number of fungal inventories has increased in recent

years (Maia et al. 2015). Several studies in different habitats in Brazil, especially

in the semiarid region, have revealed novel taxa and new records of conidial

fungi (Concei¢ao et al. 2016, Fiuza et al. 2016, Monteiro et al. 2016). During

a mycological survey of conidial fungi occurring on decaying plant material

in the Brazilian semiarid region, an interesting specimen was found, clearly

congeneric with Bharatheeya D’Souza & Bhat but different from the two

previously described species. It is therefore described here as a new species.

358 ... Conceicao & al.

Materials & methods

Samples of decaying plant material from Serra da Fumaga, Bahia, Brazil, were placed

in paper bags. The materials were washed in tap water and placed on moist filter paper

in glass Petri dishes (100 mm diam.). The dishes were then placed in plastic containers

(150 L capacity), containing 200 mL sterile water and 2 mL glycerol, and incubated

at room temperature (Castafieda-Ruiz et al. 2016). The plant material was regularly

examined over a month of incubation for the presence of microfungi. Mounts were

prepared in PVL (polyvinyl alcohol and lactic acid) and measurements were taken at

x1000. Microphotographs were made using a DP25 Olympus digital camera attached

to a BX51 Olympus microscope equipped with bright field and Nomarski interference

optics. The specimen was deposited in the Herbarium of Universidade Estadual de Feira

de Santana, Bahia, Brazil (HUEFS).

Taxonomy

Bharatheeya coronata L.B. Conc., M.F.O. Marques, J.S. Monteiro,

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

MycoBAnk MB819175

Differs from both other Bharatheeya spp. by its smooth conidiophores, its terminal

conidiogenous cells that are truncate at the apex, and its coronate, 1-distoseptate, pale

brown conidia.

Type: Brazil. Bahia State: Pindobacu, Serra da Fumaga, 10°39’S 40°22’W, on decaying

twig of unidentified plant, 8.Aug.2015, coll. L.B. Conceicao (Holotype: HUEFS 216708).

EryMo_ocy: Latin, coronata, referring to the crown-like projections on the conidial

apex.

Cotontgs on the natural substrate effuse, hairy, brown to pale brown. Mycelium

superficial and immersed, composed of branched, cylindrical, septate,

smooth, 2-3 um diam., pale brown hyphae. CONIDIOPHORES macronematous,

mononematous, single, erect, straight or slightly flexuous, unbranched,

cylindrical, 2—5-septate, thick-walled, smooth, brown, paler toward the apex,

62.5-147.5 um long, basal cell slightly inflated or sometimes radially lobed,

4-15 um diam. CONIDIOGENOUS CELLS polytretic, integrated, terminal or rarely

subterminal, cylindrical, truncate at the apex, smooth, pale brown, (9-)18-28

x 3-4 um; 30-60 um long, with or without percurrent extensions. Conidial

secession schizolytic. Conip1A solitary, acropleurogenous, 1-distoseptate,

cuneiform, turbinate to broadly pyriform, dry, smooth, pale brown, 10-17 x

7-11 um, with a small dark scar at the base, 1-2 um diam., coronate at the apex

with up to 10 projections.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. BAHIA STATE: Santa Teresinha, Serra

da Jibdia, 10°74’S 40°36’W, on decaying wood of unidentified plant, 25.11.2009, ER.

Barbosa (HUEFS 216659).

Bharatheeya coronata sp. nov. (Brazil) ... 359

Fic 1. Bharatheeya coronata: (ex HUEFS 216708). A. Conidia. B. Conidiogenous cells.

C, D. Conidiogenous cells and conidia. E. Conidiophores and conidiogenous cells.

F. Conidiophores, conidiogenous cells, and conidia;. Arrows indicate the conidiogenous cell

extensions.

360 ... Conceicao & al.

Note: Bharatheeya was established by D’Souza & Bhat (2002) and typified

by Spadicoides goanensis Bhat & W.B. Kendr. [= B. goanensis (Bhat &

W.B. Kendr.) D’Souza & Bhat]. The genus is distinguished by conidiophores

that are mononematous, smooth or verrucose, intercalary, and integrated,

polytretic conidiogenous cells, and solitary, distoseptate, smooth to rough-

walled conidia. Bharatheeya coronata shares polytretic conidial ontogeny

and distoseptate conidia with B. goanensis and B. mucoidea D’Souza & Bhat.

However, B. goanensis is distinguished by its verrucose conidiophores and

3-distoseptate conidia that are cylindrical-ovoid to obclavate (described

inaccurately as “clavate” by D’Souza & Bhat 2002), while B. mucoidea is

characterized by 2—4-distoseptate conidia (with globose, mucilaginous apical

tunica) that are obpyriform to obclavate (inaccurately as “pyriform to clavate”

in D’Souza & Bhat 2002).

Acknowledgments

The authors express their sincere gratitude to Dr. Xiu-Guo Zhang and Dr. De-Wei Li

for their critical reviews of the manuscript. LBC is grateful to the Universidade do Estado

da Bahia (UNEB), the Conselho Nacional de Desenvolvimento Cientifico e Tecnoldgico,

the Fundacao de Amparo a Pesquisa do Estado da Bahia (FAPESB) and Programa de

Pés-Graduacao em Botanica (PPGBot/ UEFS). JSM is grateful to the PROTAX N°

150823\2015-0. RFCR is indebted to the OSDE, Grupo Agricola from Cuban Ministry

of Agriculture and ‘Programa de Salud Animal y Vegetal; project P131LH003033 for

facilities. Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature

review are greatly appreciated.

Literature cited

Castafieda-Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and South

America. 197-217, in: DW Li (ed.). Biology of Microfungi. Springer International Publishing.

https://doi.org/10.1007/978-3-319-29137-6_9

Concei¢gao LB, Marques MFO, Gusmao LFP, Monteiro JS, Castafieda-Ruiz RF. 2016. Linkosia

aquatica sp. nov. from submerged plant debris from Brazil. Mycotaxon 131(2): 297-304.

https://doi.org/10.5248/131.297

D’Souza M, Bhat DJ. 2002. Bharatheeya, a new hyphomycete genus from India. Mycotaxon 83:

397-403.

Fiuza PO, Monteiro JS, Gusmao LFP, Castafieda-Ruiz RF. 2016. Mirandina uncinata sp. nov. from

submerged leaves from Brazil. Mycotaxon 131(1): 141-144. https://doi.org/10.5248/131.141.

Maia LC, Junior AAC, Cavalcanti LH et al. 2015. Diversity of Brazilian Fungi. Rodriguésia 66(4):

1033-1045. https://doi.org/10.1590/2175-7860201566407

Monteiro JS, Concei¢ao LB, Marques MFO, Gusmao LFP, Castafieda-Ruiz RF. 2016. Dictyoaquaphila

appendiculata gen. & sp. nov. from submerged wood from Brazil. Mycotaxon 131(1): 177-183.

https://doi.org/10.5248/131.177

MY COTAXON

April-June 2017— Volume 132, pp. 361-372

https://doi.org/10.5248/132.361

New records of Phallales from Paraguay

MICHELLE GERALDINE CAMPI GAONA!’, LARISSA TRIERVEILER-PEREIRA”

& YANINE ELIZABETH MAUBET CANO’

"Universidad Nacional de Asuncion, Facultad de Ciencias Exactas y Naturales,

Av. Mariscal Estigarribia Km 10, CP 2169, San Lorenzo, Paraguay

? Instituto de Botanica, Nucleo de Pesquisa em Micologia,

Av. Miguel Stéfano, 3687, Vila Agua Funda, CEP 04301-902, Sao Paulo, Brazil

* CORRESPONDENCE TO: geraldine_campi@hotmail.com

AsBstRAcT—Four phalloid fungal species collected in the Central Department are reported

for the first time from Paraguay: Clathrus columnatus, Itajahya galericulata, I. rosea, and

Mutinus argentinus. Macro- and microscopic features are described for each species and

photographs of fresh basidiomata are also provided. Additional comments address species

distribution and taxonomy.

Key worps—Clathraceae, gasteroid fungi, neotropical fungal diversity, Phallaceae, stinkhorns

Introduction

Gasteromycetes Fr. is no longer recognized as a formal class (Hibbett et al.

1997), and some of its members have been accommodated into the subclass

Phallomycetidae K. Hosaka et al. within the orders Gomphales Julich, Phallales

E. Fisch., Hysterangiales K. Hosaka & Castellano, and Geastrales K. Hosaka &

Castellano (Hosaka et al. 2006, Hibbett et al. 2007).

Within the Phallales spores are dispersed by insects, especially Diptera

(Hibbett et al., 1997); Phallaceae Corda is a well defined family that embraces

the commonly known “stinkhorns,’ which are characterized primarily by

a simple hollow pseudostipe and a slimy spore mass (glebal mass) usually

supported by a campanulate receptacle or spread over the pseudostipe surface

(Burt 1896, Lloyd 1909, Cunningham 1944, Zeller 1949, Calonge 1998).

362 ... Campi, Trierveiler-Pereira & Maubet

Several genera have been included in Phallaceae. Cunningham

(1944) accepted seven genera: Aporophallus Moller, Dictyophora Desv.,

Floccomutinus Henn., Itajahya Moller, Mutinus Fr., Phallus L., and

Staheliomyces E. Fisch. Zeller (1949) included another three: Echinophallus

Henn., Jansia Penz., and Xylophallus (Schltdl.) E. Fisch. More recently, Kreisel

(1996) treated Aporophallus, Dictyophora, and Itajahya as synonymous

with Phallus, a taxonomic treatment that has been accepted for many years

(Calonge 2005).

Although phylogenetic studies based on molecular data currently support

Dictyophora within Phallus (Hosaka et al. 2006, Trierveiler-Pereira et al.

2014), however, they also support an independent generic status for Itajahya

with two species: I. galericulata and I. rosea. Itajahya can be separated

morphologically from other phalloid genera by the presence of a discoid

structure (calyptra) that surrounds the receptacle apex and the morphology

of the receptacle (Cabral et al. 2012, Marincowitz et al. 2015). The absence of

receptacle and veil separate Mutinus species where the mucilaginous glebal

mass spreads directly on the pseudostipe surface (Calonge 1998).

For many years, Clathraceae Chevall. has been regarded as separate from

Phallaceae (e.g., Hawksworth et al. 1995: 581). Dring (1980) characterized

taxa in the Clathraceae by basidiomes bearing the mucilaginous gleba on

the inner surface of the receptacle, which may be lattice-shaped or consist of

several columns. When the gleba is exposed to the exterior, then it is located

on the surface of an apical lattice or column cluster. Although molecular

studies support Clathraceae as a monophyletic group worthy of recognition

at family level (Hosaka et al. 2006, Trierveiler-Pereira et al. 2014), Phallaceae

sensu stricto and Clathraceae are still frequently treated as a single family,

Phallaceae sensu lato (Kirk et al. 2001, 2008).

Mycological research in Paraguay was initiated by Spegazzini (1884,

1888, 1891), who reported 12 species from the country. More recent studies

have been carried out by Campi and collaborators (Campi et al. 2012, 2015;

Campi & Maubet 2015a,b). Here we report four new records of phalloid

species from Paraguay: Clathrus columnatus (Clathraceae) and Itajahya

galericulata, I. rosea, and Mutinus argentinus (Phallaceae).

Materials & methods

Specimens were collected in the Central Department, Paraguay, on the Universidad

Nacional de Asunci6n campus and in the Asuncién urban area. All specimens are

deposited at the herbarium of the Facultad de Ciencias Exactas y Naturales, Universidad

Nacional de Asuncion, Paraguay (FACEN).

Phalloid fungi new for Paraguay ... 363

Description of macroscopic features, as basidiomata size, shape and colors, were

based on fresh specimens. For the microscopic analysis portions of the basidiomata

were mounted in 5% KOH and examined with a Carl Zeiss compound microscope.

Anatomical structures were dyed with phloxine and Congo red.

Fic 1. Clathrus columnatus.

A-C. Basidioma in situ growing near Pinus. D. Immature basidioma in transverse section.

Taxonomy

Clathrus columnatus Bosc, Mag. Gesell. Naturf. Freunde, Berlin 5: 85,1811. Fre. 1

IMMATURE BASIDIOMATA epigeous, globose to ovoid, 3.2—-3.7 x 2.5-2.7

cm, attached to the substrate by rhizomorphs <7 cm long. Peridium of two

layers, the outer membranous, <0.1 cm thick, whitish, smooth, covering the

inner gelatinous layer, colorless, thickening (<4 cm thick) towards the apex

until only the exoperidium remains covering the columns of the mature

basidioma. Dehiscing by irregular tears from the apex or occasionally

364 ... Campi, Trierveiler-Pereira & Maubet

circumferentially. EXPANDED BASIDIOME 6.8-10.3 cm high, whitish at the

base, becoming yellowish to orange or reddish towards the apex, divided

into 3-4 columns of 1.4-1.9 cm diam., spongy, free at the base, attached

together at the apical zone by a transverse column. GLEBA confined in the

inner portion of the apex, olivaceous and mucilaginous, odor fetid.

BASIDIOSPORES 4-5 X 1.3—2 um, cylindrical, hyaline to greenish in KOH,

congophilic, with 1—2 internal guttules at the ends, thin-walled, smooth.

COLUMNS composed of pseudoparenchymatous, irregularly rounded cells,

16-35 um diam. EXOPERIDIUM composed of septate hyphae, 4.5-7.5 um

diam., thin-walled, branched; the inner gelatinous layer composed of

thin-walled hyphae, (2-)3-4 um diam., hyaline; septa with clamp

connections.

ECOLOGY & DISTRIBUTION—Ssolitary or in small groups, appearing

soon after rain in cold temperatures on soil among litter in a Pinus elliottii

Engelm. plantation in an urban setting.

SPECIMENS EXAMINED—PARAGUAY. CENTRAL DEPARTMENT, San _ Lorenzo,

25°20'13"S 57°31’33”'W, 26.V.2016, leg. Y. Maubet N° 16 (FACEN 3343).

DISTRIBUTION IN SOUTH AMERICA— Brazil (Trierveiler-Pereira & Baseia 2009,

Magnago et al. 2013) and Chile (Sandoval-Leiva et al. 2014). This is the first

record of C. columnatus from Paraguay.

CoMMENTS— Dring (1980) noted that a variety of forms can be found with

respect to the number of arms and the general shape of the receptacle. In

North American specimens, a transverse arm connects the vertical arms,

as in our material. Dring (1980) also observed that the type description of

Clathrus australis Speg. agrees with the type description of C. columnatus

except for the presence of a fifth column. Unfortunately, as the Argentinean

type material C. australis could not be found, some taxonomic doubt

remains regarding the separation of the two species. Spegazzini (1887a)

stated that he also had samples of C. australis from Paraguay, but they were

in very bad condition. Spegazzini (1887b) later retained both C. columnatus

and C. australis from Brazil as independent species.

Sandoval-Leiva et al. (2014) and Magnago et al. (2013) characterize the

species as having 2-5 robust spongy reddish to orange columns free at the

base and fused at the apex and a gleba that is spread on the internal portion

of the columns and not confined to a glebifer. Based on the description and

illustrations presented by previous authors (Dring 1980, Trierveiler-Pereira

2015) we conclude that our material represents C. columnatus.

Phalloid fungi new for Paraguay ... 365

Fic. 2. Itajahya galericulata. A. Basidioma in situ growing near Guadua.

B. Expanded basidioma and immature stage in transverse section.

Itajahya galericulata Maller, Bras. Pilzbl.: 79, 148, 1895. Fig. 2

IMMATURE BASIDIOMATA epigeous, subglobose to globose, 4-5 x 3.5—4

cm, exoperidium ochre to dun, slightly leathery at the apex. Basal mycelial

cord thick, 3-5 cm long and 0.2—0.4 cm thick. In transverse section, the

peridium has four distinguishable layers with the continuous mucilaginous

layer located between the other layers between the peridium and gleba.

EXPANDED BASIDIOMA phalloid in shape. Stipe white, hollow, spongy and

366 ... Campi, Trierveiler-Pereira & Maubet

externally longitudinally striated, 13.5—-20 cm high x 2-4 cm diam., tapering

towards the base. Receptacle cylindrical with grooved surface, up to 3.5

x 2.5 cm, covered with the mucilaginous gleba. GLEBA deliquescent, dark

green, odor putrid; calyptra at the receptacle apex white, discoid, flattened,

2-3 cm in diam., spongy, forming a sterile platform above the receptacle.

BASIDIOSPORES 5-6 X 2-3 tum, cylindrical, hyaline to slightly

light green in KOH, thin-walled, smooth. RECEPTACLE hyphae

pseudoparenchymatous, globose to cylindrical, 400-100 x 20-45 um;

calyptra hyphae pseudoparenchymatous (globose to cylindrical, 33-70 x

24—34 um) and thin-walled (septate, clamped, 2-13 um diam.). MyCELIAL

CORD HYPHAE thin-walled, septate, clamped, 3-5 um diam.

ECOLOGY & DISTRIBUTION—Gregarious, fruiting soon after heavy rain

during summer on soil among fallen litter and undergrowth in an urban

area.

SPECIMENS EXAMINED— PARAGUAY. CENTRAL DEPARTMENT, Asuncion, 25°18’23”S

57°35’41”W, 21.1.2014, leg. Y. Maubet N° 38 (FACEN 3424).

DISTRIBUTION IN SOUTH AMERICA—Brazil (Moller 1895, Rick 1961),

Argentina (Spegazzini 1927, Fries 1909, Wright 1949, Ruiz Leal 1954,

Dominguez de Toledo 1995, Hernandez Caffot et al. 2015), and Bolivia

(Rocabado et al. 2007). This is the first record of I. galericulata from

Paraguay.

ComMMENTS—Moller (1895) stated that in South America Itajahya

basidiomata are very variable in size and development at the edge of the

receptacle and calyptra, unlike specimens found in North America, where

they appear more stable (Long & Stouffer 1943). Dominguez de Toledo

(1995) observed that in its “egg phase” the I. galericulata peridium consists

of four layers around the gleba: two firm layers surrounding a mucilaginous

and continuous layer, the same arrangement as we found in our specimen.

We also observed longitudinal stripes in the stipe, a remarkable character

that has not been noted in the literature. The morphological feature

separating I. galericulata from I. rosea is the color of the pseudostipe, which

is pinkish in the latter.

Marincowitz et al. (2015) reported an association between I. galericulata

and the tree species Jacaranda mimosifolia D. Don in Pretoria, South Africa.

In Brazil, Bolivia, and Paraguay, specimens were collected in pastures of

urban areas in accordance with the ecology noted by Dominguez Toledo

(1995) and Hernandez Caffot et al. (2015), who described I. galericulata

growing in compacted soils with low grasses during summer and autumn.

Phalloid fungi new for Paraguay ... 367

Fic. 3. Itajahya rosea. A. Basidioma in situ growing among grass.

B. Immature stage in transverse section and expanded basidioma.

Itajahya rosea (Delile) E. Fisch., Ber. Deutsch. Bot. Ges. 47: 294, 1929 BIG.F3

IMMATURE BASIDIOMATA epigeous, subglobose to globose, 1.4-1.7

x 3-3.4 cm, exoperidium ochre to dun, slightly leathery at the apex. In

transverse section, it is possible to observe that the pseudostipe is pink and

surrounded by three layers: outer layer 0.3 cm thick, firm, white to beige,

crackled; middle layer 0.8 cm thick, mucilaginous, aqueous, hyaline; inner

layer >0.1 cm thick, whitish, dry, covering the green gleba. ExPANDED

BASIDIOMA Stipe white, hollow, fibrous, 9.5-14.2 x 1.6-1.8 cm, slightly

tapering towards the base. Receptaculum conic with reticulated surface, 2

x 1 cm, covered with gleba. GLEBA mucilaginous, deliquescent, dark green,

odor fetid; calyptra at the apex of the receptaculum white, discoid, flattened,

1.1-1.9 cm in diam., forming a sterile platform above the receptacle.

BASIDIOSPORES 4 x 2 um, cylindrical, hyaline, slightly green, thin-walled,

smooth. StrpE hyphae pseudoparenchymatous, globose, 15-25(-35) x

368 ... Campi, Trierveiler-Pereira & Maubet

22—35(-46) um. RECEPTACLE hyphae pseudoparenchymatous, globose to

cylindrical, 12-23(-26) x 8—14(-16) um; calyptra hyphae thin-walled, septate,

clamped, 3-5(-6) um diam.

ECOLOGY & DISTRIBUTION—Gregarious, appearing soon after heavy rain

during summer.

SPECIMENS EXAMINED— PARAGUAY. CENTRAL DEPARTMENT, San Lorenzo, Campus

Universitario, 26°20'08”S 57°31'14”W, 1.2016, leg. Y. Maubet N° 002 (FACEN 3338).

DISTRIBUTION IN SOUTH AMERICA—Northeastern Brazil (Ottoni et al. 2010,

Cabral et al. 2012). This is the first record of I. rosea from Paraguay.

CoMMENTS—According to Ottoni et al. (2010), the most relevant

characteristics for the taxonomical determination of I. rosea is the presence

of a calyptra at the apex of the receptacle and the pinkish pseudostipe.

The species is morphologically very close to I. galericulata (see comments

above). The peridium of the immature basidiome is another important

feature separating the species: three-layered in I. rosea and four-layered in

I. galericulata.

Moreno et al. (2013) recognized I. rosea as a rare species characteristic of

desert and semiarid regions, as reported from Yemen (Kreisel & Al-Fatimi

2008) and Northeastern Brazil (Ottoni et al. 2010). However, our specimen

was found in eastern Paraguay characterized by a subtropical climate with

the rainy season occurring in the summer.

Future molecular research is needed to separate the two morphologically

similar species.

Mutinus argentinus Speg., Anal. Soc. Cient. Argent. 24: 62, 1887. Fic. 4

IMMATURE BASIDIOMATA epigeous, ellipsoid to ovoid, 3.0 x 1.8 cm,

whitish, with basal rhizomorphs 3-6 cm long, fibrous, strongly attached to the

substratum. In transverse section releasing an odorless translucent liquid; three

layers surrounding the pseudostipe: the outer layer 0.1 cm thick, whitish to

cream, covered with brownish fibers; medium layer 0.4 cm thick, wet, sticky,

hyaline; inner layer >0.1 cm, very thin, whitish, enclosing the gleba. EXPANDED

BASIDIOME Stipe 6.6 x 0.9 cm, spongy, whitish at the base, becoming pinkish

towards the apex, and finally bright red to maroon at the apex, apex with a

small pore. GLEBA mucilaginous, olive green, covering the stipe apex.

BASIDIOSPORES 5—7 Xx 2-3 um, cylindrical, hyaline, slightly green, thin-

walled, smooth. StrpE hyphae pseudoparenchymatous, globose, 35-50 x 50—70

um, Congo-Red negative. GLEBAL REGION hyphae pseudoparenchymatous,

cylindrical, spherical to irregular in shape, 30-50 x 25-30 um, pinkish.

Phalloid fungi new for Paraguay ... 369

Fic. 4. Mutinus argentinus.

A. Basidiome in situ on dead branch, among leaves. B. Immature stage on decaying wood

and expanded basidiome and immature stage in transverse section.

ECOLOGY & DISTRIBUTION—Growing solitary or in small groups of a few

basidiomata on sandy soil among decaying fallen litter and wood debris.

SPECIMENS EXAMINED—PARAGUAY. CENTRAL DEPARTMENT, San Lorenzo, Campus

Universitario, 26°20’08”S 57°31'14”W, 23.III.2014, leg. M. Campi N° 45 (FACEN 3356).

DISTRIBUTION IN SOUTH AMERICA—Argentina (Spegazzini 1887a, 1906;

Wright 1949) and Brazil (Meijer 2006, Magnago et al. 2013). This is the first

record of M. argentinus from Paraguay.

COMMENTS—Magnago et al. (2013) discussed the similarity of M. argentinus

and M. bambusinus (Zoll.) E. Fisch., two species that have been treated as

synonyms by several authors. Dring & Rose (1977) presented the morphological

differences between the two species: M. bambusinus produces a larger fertile

portion that is strongly pigmented and with a sterile tip, while M. argentinus

lacks the marked sterile tip. The species is also depicted by Trierveiler-Pereira

(2015). Based on this information and the provisional Mutinus key by Silva et

al. (2015), we conclude that our specimen represents M. argentinus.

370 ... Campi, Trierveiler-Pereira & Maubet

Acknowledgments

The authors thank the Faculty of Natural Sciences (National University of

Asuncion) for the constant support to the research team of the Mycological Center.

We also acknowledge the contributions of the pre-submission reviewers, Dr. Orlando

Popoff (Universidad Nacional del Nordeste, Argentina) and Dr. Maria Martha Dios

(Universidad Nacional de Catamarca, Argentina). We would also like to express

immense gratitude towards Dra. Luciana Hernandez Caffot (Universidad Nacional de

Jujuy) from Argentina for the invaluable help writing the manuscript.

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MY COTAXON

April-June 2017— Volume 132, pp. 373-379

https://doi.org/10.5248/132.373

Arthrotaeniolella aquatica gen. & sp. nov. and

Pseudospiropes piatanensis sp. nov. from Brazil

JOSIANE SANTANA MONTEIRO ', LUCAS BARBOSA CONCEICAO °,

Luis FERNANDO PASCHOLATI GUSMAO * & RAFAEL F. CASTANEDA-RUIZ ”

' Universidade Estadual de Feira de Santana,

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

? Instituto de Investigaciones Fundamentales en Agricultura Tropical (INIFAT) Alejandro de Humboldt,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

* CORRESPONDENCE TO: Igusmao@uefs. br

ABSTRACT—A new genus and species, Arthrotaeniolella aquatica, and a new species,

Pseudospiropes piatanensis, collected on submerged decaying plant materials in the Brazilian

semiarid region, are described and illustrated. Arthrotaeniolella aquatica is characterized by

distinct, sinuate, slender, branched, hyaline or subhyaline to pale dirty brown conidiophores

that produce cylindrical to sub-oblong, unicellular to euseptate, dark brown conidia formed

by thallic-arthric disarticulation of the branches. Pseudospiropes piatanensis is distinguished

by a polyblastic conidiogenous cell that produces fusiform to lunate, 10-15-distoseptate, pale

brown conidia.

Key worps—Ascomycota, hyphomycetes, taxonomy, tropics

Introduction

The Brazilian semiarid region, located in the northeast of the country,

covers approximately 900,000 km?’ and has high biodiversity (Giulietti et

al. 2006). Its different vegetation types have favored a great diversity of

microfungi with the presence of several new species (e.g., Barbosa et al.

2013, Silva & Gusmao 2013, Almeida et al. 2014, Silva et al. 2014, Monteiro

et al. 2016). During continuing mycological surveys of microfungi from

this region, two interesting hyphomycetes were collected on decaying plant

materials submerged in a stream. Close examination of their morphological

374. ... Monteiro & al.

characteristics revealed that one clearly belongs to the genus Pseudospiropes

M.B. Ellis (Ellis 1971). The other, which shows remarkable differences from all

previously described hyphomycetes (Seifert et al. 2011), required the erection

of a new genus, Arthrotaeniolella.

Materials & methods

Samples of submerged litter were placed in plastic bags, taken to the laboratory,

placed in Petri dish moist chambers, and stored in a 170 L polystyrene box with 200

mL water plus 2 mL glycerol at room temperature for 30 days (Castafieda-Ruiz et

al. 2016). Mounts were prepared in PVL (polyvinyl alcohol, lactic acid, and phenol)

and measurements were made at a magnification of x1000. Microphotographs

were obtained with an Olympus BX51 microscope equipped with bright field and

Nomarski interference optics. The type specimens are deposited in the Herbarium of

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

Taxonomy

Arthrotaeniolella L.B. Conc., M.EO. Marques, J.S. Monteiro, Gusmao &

R.E. Castafieda, gen. nov.

MycoBank MB 821602

Differs from Taeniolella by its production of conidia by thallic-arthric disarticulation of

conidiophore branches.

Type Species: Arthrotaeniolella aquatica L.B. Conc. et al.

Erymo tocy: Greek, arthro-, meaning jointed, referring to the arthric-thallic conidia

formed by breaking up fertile branches at the septa; -taeniolella, referring to the genus

Taeniolella.

CONIDIOPHORES macronematous, mononematous, branched, septate,

subhyaline or pigmented. CONIDIOGENOUS HYPHAE holothallic, pigmented.

Conidial secession schizolytic. Conrp1a solitary, cylindrical to sub-oblong,

unicellular or euseptate, dark pigmented, formed by disarticulation of the

conidiogenous branches.

Arthrotaeniolella aquatica L.B. Conc., M.EO. Marques, J.S. Monteiro, Gusmao &

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

MycoBank MB 821603

Differs from Taeniolella rudis by its alternately arranged branches conidiophores and its

random thallic-arthric conidial ontogeny

Type: Brazil, Bahia State, Pindobagu, Serra da Fumaga, 40°21’W 10°39’S, on submerged

decaying leaves and twigs of unidentified plant in a stream, 26.July 2016, coll. L.B.

Conceicao (Holotype, HUEFS 224988).

EryMo_oey: Latin, aquatica, referring to its growing in water.

Arthrotaeniolella gen. nov. and Pseudospiropes sp. nov. (Brazil) ... 375

Fic. 1. Arthrotaeniolella aquatica (holotype, HUEFS 224988). A. On natural substrate; B. General aspect;

C, D. Conidia; E, F. Disarticulating conidia. G. Disarticulated conidia. Scale bars: A = 100 um; B-G = 10 um.

376 ... Monteiro & al.

COLONIES on natural substrate effuse, hairy, brown. MyceL1um mostly

superficial, hyphae branched, septate, 1-2 um diam., smooth, subhyaline

to pale dirty brown. CONIDIOPHORES macronematous, mononematous,

slender, erect, flexuous or straight, subhyaline or pale dirty brown,

55-250 x 6-9 um; sometimes unbranched but mostly with dark dirty brown,

4—7-septate, 40-80 um long branches, alternately arranged toward the apex

CONIDIOGENOUS HYPHAE holothallic, branches and differentiated apical

region of the unbranched conidiophores cylindrical to long clavate, mostly

discrete, sometimes integrated, dark dirty brown. Conrp1a cylindrical,

sub-oblong, truncated at the ends, occasionally rounded at the apex,

0-4-septate, 8-35(-40) x 3-9 um, dark dirty brown, dry, smooth, forming

by random thallic-arthric disarticulation at the septa of the conidiogenous

hyphae.

Note: Taeniolella S. Hughes is characterized by conidiophores that are

little differentiated from hyphae, usually short, unbranched or sparingly

branched near the base and conidiogenous cells that are monoblastic,

integrated, terminal, cylindrical or doliiform and that produce conidia

that are solitary or in branched acropetal chains, 1 or more septate, often

seceding with difficulty, cylindrical, dark brown (Hughes 1958, Ellis 1971,

Seifert et al. 2011). In T! rudis (Sacc.) S. Hughes the conidial chains are

broken up by disarticulation (Jones et al. 2002; Seifert et al. 2011). In

Arthrotaeniolella the conidiogenous events occurring in the branches and

hyphae are classified as conidial development type 38 (conidial ontogeny

holothallic, conidiogenous cells formed by apical wall-building coincident

with conidial ontogeny, random delimitation by 1 septum at each end,

no maturation during conidiogenesis, secession randomly schizolytic) in

Kirk et al. (2008), which is different from the blastic conidial ontogeny in

Taeniolella.

Two other genera illustrated by Seifert et al. (2011)—Ampulliferina

B. Sutton and Thielaviopsis Went—have similar thallic-arthric conidial

development in the branches. In Ampulliferina the superficial hyphae

are hyphopodiate, with lateral, globose, clavate, rarely lobate, brown

hyphopodia (Sutton 1969, Seifert et al. 2011). In Thielaviopsis the thallic-

arthric development is restricted to the chlamydospores-like branches,

which are distinguished by the production of unicellular or septate, dark

brown arthroconidia with a germ slit or pore (Seifert et al. 2011).

Arthrotaeniolella gen. nov. and Pseudospiropes sp. nov. (Brazil) ... 377

Fic. 2. Pseudospiropes piatanensis (holotype, HUEFS 211342). A. Conidia; B-D. Conidiophores;

E. Detail of conidiogenous cell; F. Conidiogenous cell and conidia. Scale bars = 10 um.

Pseudospiropes piatanensis J.S. Monteiro, Gusmao & R.F. Castafieda, sp. nov. Fic. 2

MycoBank MB 821604

Differs from Pseudospiropes mussaendae and P. ximeniae by its 10-15-distoseptate

fusiform to lunate conidia.

378 ... Monteiro & al.

Type: Brazil, Bahia State: Piata, Rio Coxd, 13°00’S 41°53’W, on submerged decaying

wood in a stream, 9 May 2014, coll. J.S. Monteiro (Holotype, HUEFS 211342).

ETYMOLOGY: piatanensis, referring to the City of Piata where the type was found.

COLONIES on natural substrate effuse, hairy, brown. Mycelium mostly

immersed, hyphae branched, septate, smooth, pale brown, 1-2 um diam.

CONIDIOPHORES macronematous, mononematous, single, simple, erect,

straight or slightly flexuous, septate, smooth, dark brown, pale brown toward

the apex, 82.5-87.5 x 4-5 um. CONIDIOGENOUS CELLS holoblastic, polyblastic,

indeterminate, integrated, terminal (becoming intercalary), sympodial,

cylindrical, smooth, pale brown, 27.5-52.5 x 6-7.5 um. Conidiogenous loci

enlarged, thickened, protuberant, melanized, lenticular. Conidial secession

schizolytic. Conip1A solitary, acropleurogenous, fusiform to lunate,

10-15-distoseptate, smooth, pale brown, 57.5-72.5 x 10-16 um, 2.5-3 um

diam. at the truncate base.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, Banta: Piataé, Rio Cox, 13°00’S

41°53’W, on submerged decaying wood in a stream, 9 May 2014, coll. J.S. Monteiro

(HUEFS 211343).

Note: Pseudospiropes is distinguished by solitary, acropleurogenous,

distoseptate conidia seceding schizolytically from polyblastic, sympodial,

cicatrized conidiogenous cells with enlarged, thickened, protuberant and

melanized conidiogenous loci (Ellis 1971, Castafieda-Ruiz et al. 2001, Seifert

et al. 2011). The 17 species currently included in this genus are delimited by

conidial shape, size, number of septa, and pigmentation (Castaneda et al.

2001, Ma etal. 2011). Among Pseudospiropes species, only P mussaendae Z.Q.

Shang & X.G. Zhang and P. ximeniae Z.Q. Shang & X.G. Zhang have conidia

similar to those of P. piatanensis. However, the conidia of P. piatanensis have

more distosepta and are longer than those of P mussaendae (conidia 36-47

x 9-11 um, 8-9-distoseptate) and P ximeniae (conidia 34-40 x 12-14 um,

6-8-distoseptate). In addition, the conidia of P piatanensis are fusiform to

lunate, while those of P mussaendae are fusiform to obclavate and those of

P. ximeniae fusiform to navicular (Shang & Zhang 2007).

Acknowledgments

We are indebted to Dr. Jian Ma and Dr. De-Wei Li for critical review. We are grateful

to Dr. Keith A. Seifert for his opinion and commentaries. The authors thank the National

Council for Scientific and Technological Development (CNPq) (Proc. 451991/2016-8).

The first author is grateful to Coordination for the Improvement of Higher Education

Personnel (CAPES) through project N° 071/2012. RFCR is grateful to OSDE, Grupo

Agricola from Cuban Ministry of Agriculture and Programa de Salud Animal y Vegetal

Arthrotaeniolella gen. nov. and Pseudospiropes sp. nov. (Brazil) ... 379

(project P131LH003033) for facilities. Dr. Lorelei L. Norvell’s editorial review and

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

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fungi from the semi-arid Caatinga biome of Brazil. Nova Hedwigia 98: 431-447.

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nov. with secondary sympodioconidia from freshwater habitats. Mycoscience 43: 201—206.

https://doi.org/10.1007/S 102670200029

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

10th edition, CAB International, Oxon, UK. 771 p.

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and notes on Minimelanolocus (both anamorphic Strossmayeria) new to China. Nova Hedwigia

93(3-4): 465-473. https://doi.org/10.1127/0029-5035/2011/0093-0465

Monteiro JS, Gusmao LFP, Castafeda-Ruiz RF. 2016. Pleurothecium bicoloratum & Sporidesmiopsis

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

CBS Biodiversity Series 9. 997 p.

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MY COTAXON

April-June 2017— Volume 132, pp. 381-389

https://doi.org/10.5248/132.381

Cunninghamella clavata from Brazil:

a new record for the western hemisphere

ANA L.S. DE M. ALVES, CARLOS A.F. DE SOUZA, RAFAEL J.V. DE OLIVEIRA,

THALLINE R.L. CORDEIRO & ANDRE L.C.M. DE A. SANTIAGO"

Post-graduate course in Fungal Biology, Federal University of Pernambuco,

Av. Prof. Moraes Rego, s/n, 50670-420, Recife, PE, Brazil

* CORRESPONDENCE TO: andrelcabral@msn.com

AsBstTRACT— During a survey of Mucorales in Brejo dos Cavalos (a fragment of an Upland

Atlantic Forest within the semi-arid region of Northeast Brazil) a specimen of Cunninghamella

clavata was isolated from soil samples. A detailed description of the specimen and a

phylogenetic analysis of its relationship with other Cunninghamella species are presented. An

identification key for Cunninghamella taxa reported from Brazil is also provided.

Key worps—Mucoromycota, Cunninghamellaceae, LSU rDNA

Introduction

Cunninghamella Matr. (Cunninghamellaceae Naumov ex R.K. Benj.

Mucorales Fr.) comprises species characterized by the production of sporophores

with pedicellate unispored sporangiola covered with spines (Zheng & Chen

2001). The sporophores are irregularly, verticillately, or pseudoverticillately

branched, and the Mucor-like zygospores present a reddish-brown wall (Baijal

& Mehrotra 1980, Benny 2006). Cunninghamella species have a worldwide

distribution, and most are saprobes, often found in soil, stored grains, and

other organic substrates (Baijal & Mehrotra 1980, Yu et al. 2015). Some species

are pathogenic to humans, particularly immunocompromised patients (Hsieh

et al. 2013).

Species of Cunninghamella have traditionally been distinguished based on

morphological features, such as colony color and texture, pattern of sporophore

382 ... Alves & al.

branching, vesicle shape and size, sporangiola shape, size, and color, and the

presence/absence and length of the projections (spines) in the sporangiola

(Zheng & Chen 2001). Cunninghamella has been monographed by several

authors (Alcorn & Yeager 1938, Cutter 1946, Milko & Beljakova 1967, Samson

1969, Baijal & Mehrotra 1980, Lunn & Shipton 1983) based on morphological

characteristics. A subsequent monograph by Zheng & Chen (2001) based

on morphological characteristics, maximum growth temperatures, mating

experiments, and the length of the ITS region (RFLP analysis) recognized

twelve species and three varieties: C. bertholletiae Stadel, C. blakesleeana Lendn..,

C. binariae R.Y. Zheng, C. clavata, C. echinulata var. antarctica (Caretta &

Piont.) R.Y. Zheng & G.Q. Chen, C. echinulata (Thaxt.) Thaxt. ex Blakeslee var.

echinulata, C. echinulata var. nodosa R.Y. Zheng, C. echinulata var. verticillata

(FS. Paine) R.Y. Zheng & G.Q. Chen, C. elegans Lendn., C. homothallica

Komin. & Tubaki, C. intermedia K.B. Deshp. & Mantri, C. multiverticillata

R.Y. Zheng & G.Q. Chen, C. phaeospora Boedijn, C. septata R.Y. Zheng, and

C. vesiculosa P.C. Misra. Liu et al. (2001) and Yu et al. (2015), who sequenced

the ITS rDNA and tef-la gene regions of taxa accepted by Zheng & Chen

(2001), found a phylogeny that was consistent with Zheng & Chen's taxonomic

conclusions. Subsequently, two additional species have been described:

C. bigelovii Z. Xin et al. from China (Guo et al. 2015) and C. gigacellularis A.L.

Santiago et al. from Brazil (Santiago et al. 2016).

Cunninghamella clavata has previously been reported only from Guizhou

and Yunnan in southwest China, where the holotype and two paratype

specimens were collected in 1985-92 (Zheng & Chen 1998). Here we present a

detailed description of a C. clavata collection isolated from soil samples from

an upland forest area in the semiarid region of Brazil. An identification key for

Cunninghamella taxa recorded in Brazil and results of phylogenetic analysis of

Cunninghamella LSU rDNA sequences are also provided.

Material & methods

SOIL COLLECTION—Samples of soil were collected in September 2014 in Brazil’s Brejo

dos Cavalos Upland Atlantic Forest (8°16’S 35°58’W), located in the city of Caruaru

in Pernambuco state, 130 km from Recife. The mean annual temperature is 22 °C, with

rainfall ranging from 650 to 800 mm per year. The vegetation consists of montane

ombrophilous forest, a remnant of the Atlantic Rainforest. The samples were placed in

clean plastic bags and stored in Styrofoam boxes for transportation to the laboratory.

ISOLATION, PURIFICATION, AND IDENTIFICATION OF C. CLAVATA—Portions of 5 mg

of soil were added to Petri dishes containing a wheat germ agar culture medium

Cunninghamella clavata in Brazil ... 383

Absidia californica JN206583

1.00/100

A. caatinguensis KT308170

1.00/100}

A. caatinguensis KT308171

Cunninghamella vesiculosa HM849693

C. echinulata JN206598

0.99/95

1.00/400 A C. echinulata var. antarctica JN206597

C. echinulata HM849702

0.95/100 ia

C. intermedia JN206606

C.ETBE C. homothallica JN206605

C. elegans HM849700

C. bertholletiae JN206599

0.76/68,

~ + C. bertholletiae JN206600

1.00/99 i

0.85/68 C. bertholletiae HM849701

0.72/89 C. blakesleeana JN206601

0.98/89 é C. phaeospora HM849697

C. binariae JN206603

1.00/100 C. binariae JN206602

C. clavata KX369610

v4 0.85/96 C. clavata JN206604

Fic. 1. Phylogenetic tree of Cunninghamella taxa, constructed using the large subunit (LSU) rDNA

sequences, with Absidia californica and A. caatinguensis as outgroup. Sequences are labeled with

their GenBank accession numbers. Support values are from Bayesian inference and maximum

likelihood analyses. The sequence obtained in this study is in boldface.

384 ... Alves & al.

(Benny 2008) augmented with chloramphenicol (NeoFenicol—Neo Quimica;

100 mg.L"'). The dishes were stored on the laboratory bench at room temperature (26

+ 2 °C) for seven days and subjected to alternate light and dark periods. Fragments

of mycelium were removed from the Petri dishes under a stereomicroscope (Leica

EZ4) and transferred to Petri dishes with malt extract agar (MEA) (Benny 2008). The

specimen was identified by comparing the macroscopic (colony color, appearance,

and diameter) and microscopic (e.g., shape and size of sporophores, columellae,

sporangia, and sporangiospores) characteristics with the descriptions of Zheng &

Chen (1998, 2001). A voucher was conserved in the Micoteca, Federal University of

Pernambuco, Recife, Brazil (URM).

MOLECULAR ANALYsiIs—cultures grown in test tubes containing MEA were incubated

at 28 °C for six days to obtain fungal biomass. After the material was transferred to 2 mL

microtubes with screw caps, 0.5 g acid-washed glass beads of two different diameters

(150-212 um and 425-600 um, 1:1; Sigma, USA) were added to each tube. The

material was crushed through stirring at high speed in a FastPrep homogenizer. The

genomic DNA was extracted as described by Gées-Neto et al. (2005). The mycelium

was washed with chloroform: isoamyl alcohol (24:1) and then homogenized in 2%

cetyltrimethylammonium bromide buffer. The DNA was precipitated in isopropanol,

washed with 70% ethanol, and resuspended in 50 uL of ultrapure water.

The primer pairs LR1/LSU2 were used for the amplification of the large subunit

(LSU) of nuclear ribosomal DNA (rDNA) (Van Tuinen et al. 1998, Santiago et al.

2014). The polymerase chain reactions were carried out as described by Oliveira et al.

(2014). The newly obtained sequences were deposited in GenBank database.

Phylogenetic reconstructions were obtained by analyzing the sequence data of the

partial LSU rDNA gene. The fungal sequences were aligned with ClustalX (Larkin

et al. 2007) and edited with BioEdit (Hall 1999). Prior to phylogenetic analysis, the

nucleotide substitution optimal model was estimated using TOPALi 2.5 (Milne et al.

2004). Bayesian inference (two runs over 1 x 10° generations with a burn-in of 2500)

and maximum likelihood (with support estimated by a bootstrap analysis with 1000

replicates) analyses were performed with MrBayes 3.1.2 (Ronquist & Huelsenbeck

2003) and PhyML (Guindon & Gascuel 2003), launched from TOPALi 2.5.

Phylogenetic analysis

In BLASTn analysis the C. clavata sequence from our collection (URM

7410, KX369610) showed 98% identity with the sequence of the C. clavata

type culture (CBS 100178, JN206604); these two sequences nested together

in the phylogenetic tree (Fic. 1).

Taxonomy

Cunninghamella clavata R.Y. Zheng & G.Q. Chen, Mycotaxon 69: 189 (1998) Fic. 2

CoLony initially white; after 4 days in MEA at 25 °C becoming pale

Cunninghamella clavata in Brazil ... 385

| Pra RR aCe er

Fic. 2. Cunninghamella clavata (URM 7410): A. Repeatedly branched sporophore; B. Sporophore

simple with sporangiola; C, D. Sporophores with lateral branches and vesicles; E. Unbranched

sporophore with a vesicle; F Sporangiola.

grayish with cream reverse and reaching 9.4 cm diam. and 9 mm high.

RuIzo1ps abundant, long and short, simple or slightly branched. STOLONS

present, coenocytic, some showing septa near the sporophore origin.

SPOROPHORES short and long, erect or recumbent, arising from stolons or

directly from aerial hyphae, hyaline, smooth-walled; some containing one

or more septa below the sporangium or near the basis. Main sporophore

axes simple or branched, usually equal in width throughout, some enlarging

in the apical direction and ending in a vesicle (3.5—)5-10(-14) um diam.;

swelling below the vesicle present. Primary branches simple, in pairs,

386 ... Alves & al.

pseudo-verticillate (<4). Short and long branches in the same sporophore

common, repeatedly (<6x) branching, some arising close to the vesicles and

growing parallel to the main sporophores <500 x 7.5 um, erect or slightly

curved and terminating in a vesicle. VEsSICLEs of the main sporophores light

gray, mostly clavate, commonly irregular, angular, or flattened at the apex,

rarely globose or subglobose, 17-47.5 x 20-37.5 tm. LATERAL VESICLES

with same shape as the main vesicles, (2.5-)7.5-35 x (2.5)5.5-20(-25) um.

SPORANGIOLA globose, light yellow with greenish yellow content and

echinulate, (6.5—) 10-20(-23) um diam. SPORANGIOSPORES hyaline, echinulate-

walled, usually remaining within the sporangiola. CHLAMyDOsSPORES and

ZYGOSPORANGIA not observed.

Hapsirtat: isolated from Upland Atlantic Forest soil from the semiarid

region of Pernambuco, northeastern Brazil.

SPECIMEN EXAMINED: BRAZIL, PERNAMBUCO, Caruaru: Brejo dos Cavalos, (8°16’S

35°58’ W), in soil, 29.01.2015, leg. A.L. Alves (URM 7410; GenBank KX369610).

GEOGRAPHIC DISTRIBUTION: Guizhou and Yunnan, China (Zheng & Chen

1998) and Pernambuco, Brazil. This is the first report of C. clavata in the

western hemisphere.

Discussion

Alvarez et al. (2011) and Walther et al. (2013) noted that the LSU rDNA

region is a reliable indication of taxonomic differentiation in Mucorales. Our

phylogenetic and morphological analyses support C. clavata as distinct from

the other described Cunninghamella species. Morphologically, C. clavata is

distinguished by producing clavate vesicles and the formation of repeatedly

branching short and long branches within the same sporophore—some

arising close to the vesicles and growing parallel to the main sporophores

(Zheng & Chen 1998).

Cunninghamella echinulata var. echinulata also presents clavate vesicles,

but lacks the exclusively globose sporangiola observed in C. clavata, and in

C. echinulata var. echinulata colonies are white to pale, not pale grayish as

in C. clavata (Zheng & Chen 2001).

Cunninghamella intermedia also produces strictly globose sporangiola,

but its sporophores are not repeatedly branched and the vesicles are globose

to subglobose, never clavate. The phylogenetic analysis of the LSU rDNA

region confirmed the identity of our specimen.

Our Brazilian specimen represents the first western hemisphere record of

C. clavata previously reported only from southwest China (Zheng & Chen

Cunninghamella clavata in Brazil ... 387

1998), contributing to our knowledge of Mucorales distribution. In Brazil,

other species of Cunninghamella such as C. bertholletiae, C. blakesleeana,

C. echinulata var. echinulata, C. echinulata var. verticillata, C. elegans, and

C. phaeospora have been isolated from soil and from herbivore dung

(Trufem 1981; Santiago et al. 2011, 2013).

Key to Cunninghamella taxa registered in Brazil

1. Colonies white to pale or turning yellowish;

siant:dark.sporangiold present OF NOL sic oes ni ey Nino Cb Nino Eb Miso EN wins EN wine 6 2

1. Colonies pale grayish to deep or brownish gray;

siant-dark sporangiola not present. t+ ii mo cue oe aloe seedy Moers Heese Mees 4

2. Colonies floccose, pale yellowish; sporangiola ovoid or ellipsoid (<30 x 15 um)

or globose (<17 um diam.); giant dark sporangiola not present;

Inaximnum -srowth-temperature: 38 C.F Akg ee a Mee ag C. blakesleeana

2. Colony floccose or granulate, white to pale; sporangiola <22 x 18 um;

giant dark sporangiola present; maximum growth temperature 42 °C ........ 3

3. Colony floccose; sporophores usually branched (pseudo)verticillately

with shorter branches on the same sporophore rarely re-branching

mill aceon cece AiaMicil Personal Pena stn at ee C. echinulata var. verticillata

3. Colony granulate; sporophores never (pseudo)verticillately branched;

with shorter branches on the same sporophore often re-branching

Ea! seat abe seats inate hadrons ah: boskua we onli: frsetia ts eet C. echinulata var. echinulata

4, Main sporophore axis terminating in a vesicle; long and short branches

on the same sporophore present or absent, if present re-branching <6x ...... 5

4. Sporophores terminating in a vesicle or dividing into apical branches;

long and short branches in the same sporophore common,

FE DVANCNIAS SIS hy eBags ety es ck Pye a eh le oe te ee C. bertholletiae

5. Vesicles clavate; short and long branches in the same sporophore

fepedted habranchime oxi te rier tor Leen Onee tn gt eho ee eRe. Get C. clavata

5. Vesicles globose, subglobose, ovoid or slightly angular;

sporophore branches rare or common; when present branches

similar inssize and-farely-fe-Dranichin ey: 5 wise Nees woes es Mire Niorionds Mees ona Mlle 6

6. Sporophores infrequent, poorly branched;

sporangiola brownish to dark brown ............... 0.00. ee eee C. phaeospora

6. Sporophores common, well branched;

spotansiola-orayish-tO DTOWDISN aso hac a ale cal A ne A ale RY C. elegans

Acknowledgments

The authors would like to thank the Coordenacao de Aperfeicoamento de Pessoal

de Nivel Superior (CAPES) and Fundacao de Amparo a Ciéncia e Tecnologia do

Estado de Pernambuco (FACEPE) for the postgraduate scholarships to Ana L. S. M.

388 ... Alves & al.

Alves and Carlos A.F. de Souza. We would also like to thank the Conselho Nacional de

Desenvolvimento Cientifico e Tecnoldgico (CNPq) and FACEPE for financial support

throughout the projects: ‘Mucoromycotina in upland forests from the semi-arid

region of Pernambuco’ (CNPq - 458391/2014-0), and ‘Diversity of Mucoromycotina

in different ecosystems of the Pernambuco Atlantic Rainforest’ (FACEPE - APQ 0842-

2.12/14). Drs. José Ivanildo de Souza and José Luiz Bezerra are gratefully acknowledged

for helpful presubmission reviews.

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MY COTAXON

April-June 2017— Volume 132, pp. 391-419

https://doi.org/10.5248/132.391

A comparative SEM study of morphological characters

in Cribraria

JosE MARTIN RAMfREZ-ORTEGA **, ARTURO ESTRADA-TORRES ”

& EFRAIN DE LUNA 3

' Posgrado en Ciencias Bioldgicas & * Centro Tlaxcala de Biologia de la Conducta,

Universidad Autonoma de Tlaxcala,

Carr. Tlaxcala-Puebla Km. 1.5, Tlaxcala, Tlaxcala, 90070, México

> Dpto. De Biodiversidad y Sistematica, Instituto de Ecologia A. C.

Km. 2.4 Carr. Antigua a Coatepec 351 Congregacion el Haya,

Xalapa, Veracruz, 91070, México

* CORRESPONDENCE TO: jmartini.ro@gmail.com

ABSTRACT—Cribraria spp. were examined by SEM to determine detailed morphological

features of the spore, nodes, and calyculus and to identify and describe the characters and

character states for each microstructure examined. The character states defined include

six for spore ornamentation (warted, crested, spiny, subreticulate, banded-reticulate,

grooved-reticulate), three for node types (rounded, enlarged, irregular), and four for the

upper calyculus margin (ribbed, irregular, toothed, entire). Our SEM studies show that

interpretations of spore ornamentation character states based solely on LM may be incorrect

and that SEM observations are crucial for discovering new characters and interpreting

taxonomic morphological features.

Key worps—Cribrariaceae, morphology, myxomycetes, plasmodial granules, slime molds

Introduction

Myxomycetes (now formally classified by Wwww.indexfungorum.org

in Myxogastrea) are a small, relatively homogeneous group of eukaryotic

organisms with small fructifications (sporangia) and a spore size range of

5-15 um (Stephenson & Stempen 1994). The ornamented spore surface is very

difficult to assess using only light microscopy (LM) (Singer et al. 2006), and

the magnification and resolution limitations of LM prevent close examination

392 ... Ramirez-Ortega, Estrada-Torres & De Luna

of the fine structure of sporangia (Hatano 1985). Lane (1985) proposed that

scanning electron microscope (SEM) techniques be used to describe new

characters and to assist in the proper interpretation and taxonomic use of

previously unknown features.

Spore ornamentation, size, and (in some species) the way in which

the dehydrated spore wall collapses are among the most useful diagnostic

characters for identifying myxomycete species. The ability of SEM to produce

taxonomically significant information and to resolve minute details has proved

superior to other forms of microscopy in visualizing important diagnostic

characters (Schoknecht & Small 1972, Lyon & Kramer 1977). Although Gaither

& Collins (1984) credited Schoknecht & Small as the first to emphasize SEM for

resolving taxonomical questions in myxomycete systematics, Keller & Reynolds

(1971) had previously described a new species of Perichaena with the aid of the

SEM images. Subsequently several other authors have used SEM techniques to

identify new taxonomic characters (Ellis et al. 1973, Rammeloo 1974, Eliasson

& Sunhede 1980, Nelson et al. 1982) and to provide more inclusive descriptions

of new species (Keller & Reynolds 1971, Keller et al. 1975, Mulleavy 1977, Raub

et al. 1979, Whitney & Olive 1983).

Cribraria species produce some of the smaller sporangia (up to 1 mm

in C. zonatispora), which in some instances are detectable only when the

substratum is observed microscopically at 50-100x or when the specimens

are produced in moist chamber cultures. Lado (2017) currently recognizes 46

species of Cribraria. Morphological characters that vary among species—type

of nodes, surface spore ornamentation, calyculus margin, and nature of the

peridial net—are among those most frequently used in taxonomic descriptions.

In the literature, most species descriptions are based on LM observations; to

date only 18 Cribraria species have been described with SEM details (Hatano

1985, Keller et al. 1988, Lado et al. 1999, Estrada-Torres et al. 2001, Moreno

et al. 2001, Hatano 2002, Liu & Chang 2007, Hooff 2009). Hatano (1985), in a

study of myxomycetes from Japan, provided SEM details for ten species in the

Cribrariaceae (Cribraria argillacea, C. aurantiaca, C. cancellata, C. cribrarioides,

C. intricata, C. languescens, C. microcarpa, C. minutissima, C. rufa, and C.

tenella), although the spores were collapsed in C. minutissima and C. rufa; Keller

et al. (1988), who discussed the systematics of C. minutissima and C. confusa,

included SEM photographs with detailed spore morphology and noted that the

spores tended to collapse along preformed lines over the surface. Lado et al.

(1999) described C. zonatispora as having ‘unique’ spores, while Estrada-Torres

et al. (2001) described unique spore ornamentation for their new species,

SEM comparisons in Cribaria ... 393

C. fragilis, from Mexico. In their contribution to the myxomycetes of Baja

California (México), Moreno et al. (2001) described the spore ornamentation

as observed under SEM for C. intricata, C. purpurea, and C. violacea. Hatano

(2002) studied and described the surface ornamentation of the sporangium,

calyculus, peridial surface net and spore ornamentation for five species from

the Korean Peninsula (C. intricata, C. microcarpa, C. rufa, C. splendens and

C. tenella). More recently, Liu & Chang (2007) described C. angulospora and

Hooff (2009) described C. tecta using characters observed under SEM.

In a general spore classification scheme for all myxomycetes, Hatano

(1985) proposed seven types of spore ornamentation, including Cribraria in

type I (warts) with four subtypes (a, b, c, d). Each subtype was defined based

on the frequency or distribution of warts on the spore surface. Although he

considered warts the predominant type of ornamentation in Cribraria spores,

a completely different surface ornamentation has been observed under SEM in

some Cribraria species, e.g. C. zonatispora, C. fragilis, and, C. tecta (Lado et al.

1999, Estrada-Torres et al. 2001, Hooff 2009).

In view of the morphological variability of the spore surface and other

taxonomically important structures found in Cribraria, the objective of our

study was to contribute to knowledge of the genus through SEM examination

and to identify potentially informative morphological characters and character

states of the microstructures that will eventually help reveal phylogenetic

relationships within the genus.

Materials & methods

Taxon sampling

We obtained specimens representing 35 (of 46 known) Cribraria species from the

US. National Fungus Collection, Beltsville MD, U.S.A. (BPI), the herbarium of the

National Museum of Nature and Science, Tsukuba, Japan (TINS, now donated to herb

TLXM), and the myxomycete collection at the herbarium of the Universidad Autonoma

de Tlaxcala, México (TLXM). Whenever more than two specimens per species were

obtained, we processed the most abundant and typical (based on our comparison

with the descriptions) collection for SEM observation. Here we describe only SEM

observations for 21 species, as we were unsuccessful in processing the specimens

representing the other 14 species.

Processing of the sporangium for SEM

Because material collected in the field is generally dehydrated when preserved

in a herbarium, preparation for SEM observation required rehydration of the

samples for eight hours in a 1:1 solution of concentrated ammonium hydroxide

and distilled water. After rehydration, samples were dehydrated in a graded

394 ... Ramirez-Ortega, Estrada-Torres & De Luna

TRANSVERSAL

PERIDIAL

RIBS

FiGurE 1. Morphological sporocarp structures described for Cribraria species.

ethyl alcohol series of 10, 20, 30, 50, 90 and 100% concentrations and then

acetone solution for ten minutes at each step. The samples were then dried

using a critical point technique, mounted on specimen stubs with double face

adhesive tape, and coated with gold/ palladium in a high-vacuum chamber

with a JFC-1100 Jeol Fine Coat Ion Sputter. The samples were examined with a

JEOL JSM-5600 scanning electron microscope.

Results

The characters targeted under the SEM include (1) the peridium, (2) the

type of node, (3) the margin and outer surface of the calyculus, and (4) the

spore surface. Below, we detail the taxonomic characters for Cribraria as

observed under SEM and briefly compare them with the characters described

by previous authors (based on SEM and LM). For a better understanding of the

morphological terms used in this paper, we also provide a drawing depicting

important generic features in Cribraria (Fic. 1) and a brief glossary (p. 419).

Species & SEM descriptions

Cribraria atrofusca G.W. Martin & Lovejoy,

J. Wash. Acad. Sci. 22: 92, 1932. PLATE 1a—d

SPECIMEN BPI 828412—PeERIpIvm: consisting of an upper net and a shallow

cup at the base of the sporotheca. NopEs: scarcely thickened, mostly elongated,

connected by broad threads, sometimes with 2-3 free ends and forming angular

isodiametric meshes. PLASMODIAL GRANULES: concentrated principally in

SEM comparisons in Cribaria ... 395

the nodes but also in the inner side of the threads. CALycuLus: membranous,

shallow, usually well developed, about one third the total height of the entire

sporotheca, the margin bearing fine long tooth-like projections, the outer

calyculus surface characterized by pronounced concentric folds. SpoREs: (SEM

images slightly collapsed) surface ornamentation apparently composed of a

reticulum formed by thick ribs, with 4-5 meshes on one hemisphere and with

warts, sometimes fused and forming ridges, in the inner areas formed by the

meshes of the reticulum.

ComMENTs: The microstructure under SEM has not been previously

reported for C. atrofusca. Martin & Alexopoulos (1969) described the spore

ornamentation as “finely verrucose with faint, broken reticulate pattern” as seen

by LM. Although most spores were collapsed in our study, we did observe that

a ribbed-reticulate pattern and warted spore surface occur in this species. The

concentric folds in the calyculus and the peridial net are the most distinctive

features of this species.

Cribraria aurantiaca Schrad. Nov. Gen. Pl.: 5. 1797. PLATE 1e-h

SPECIMEN TNS 2612—PeEripivum: partially fugacious, consisting of a net

occupying most of the sporotheca and leaving a calyculus in the base. NoDEs:

isodiametric, most rounded, thickened, connected by threads (c. 2 um diam.)

forming a regular mesh. PLASMODIAL GRANULES: (c. 2 um diam.) concentrated

in the nodes. CaLycuLus: membranous, shallow, usually well developed,

about one-third the total height of sporotheca, margin characterized by ribs

connecting with peridial net, upper surface with delicate concentric folds,

radial folds at the base of the calyculus. Spores: globose, 5-6 um diam. with

a surface ornamentation formed by warts; frequently several fuse to form an

irregular ridge.

ComMENTs: A SEM study by Hatano (1985) reported C. aurantiaca with a spore

ornamentation composed of many warts, sometimes adhering to each other

and forming irregular ridges. Our observations of the spore ornamentation,

node, and calyculus characters agree with Hatano.

Cribraria cancellata (Batsch) Nann.-Bremek.,

Nederlandse Myxomyceten 92. 1975. PLATE 2a-c

SPECIMEN TLXM Rodriguez-Palma 2191—PeEripium: partly evanescent,

remaining as 50-60 ribs connected by fine transverse threads. PLASMODIAL

GRANULES: distributed along the rib surfaces, 1-2 um diam. Spores: globose, c.

4-5 um diam.; surface ornamentation composed of winding crests, sometimes

396 ... Ramirez-Ortega, Estrada-Torres & De Luna

branched and forming a subreticulate pattern. STaLK: grooved and attenuate

at the apex. SPOROTHECA: oblate, sometimes globose, usually umbilicate at the

apex, generally nodding, rarely erect.

COMMENTS: Martin & Alexopoulos (1969) and Lado & Pando (1997) described

the spore surface as smooth or minutely punctate, but irregular and winding

crests are easily seen with SEM. Hatano (1985), who described the spore

ornamentation in the same way, referred to C. cancellata as having vein-like

warts. We did not see a basal disc or calyculus in our specimen but did observe

that the ribs extend from the stalk apex.

Cribraria costata Dhillon & Nann.-Bremek.,

Proc. Kon. Ned. Akad. Wetensch., C 81: 141. 1978. PLATE 2d-h

SPECIMEN TNS 16898—PERIpDIUM: partially evanescent, remaining as

ribs that arise from the base to almost the top of the sporotheca, connected

by perpendicular threads, forming a net at the upper part of the sporangium.

NopeEs: pulvinate, isodiametric, connected by strip-like threads that form an

open mesh and with few free ends. PLASMODIAL GRANULES: concentrated in

the nodes. CALycutus: (if present) a basal disc. SpoREs: globose, 6-7 Um diam.;

surface ornamentation composed of winding crestae, sometimes branched and

forming a subreticulate pattern.

ComMENTs: The specimen we studied fits well the original description given

by Dhillon & Nannenga-Bremekamp (1978). The microstructure as seen by

SEM has never been reported before for C. costata. The ribs radiating from the

apex of the stalk and the pyriform shape of the sporotheca are the distinctive

characters for this species.

Cribraria dictyospora G.W. Martin & Lovejoy,

J. Wash. Acad. Sci. 22: 91. 1932. PLATE 3a—d

SPECIMEN TNS 2936—PERIDIUM: with a shallow basal cup anda fine meshed

net in the upper portion of the sporotheca. Nopes: flattened and elongated,

with narrow connecting threads with few free ends. CaLycutus: shallow,

occupying one-third the total height of the sporotheca, marked with radial

folds on the outer surface, margin toothed. Sporzs: polyhedric, 5-6 um diam.,

surface ornamentation composed of a wide meshed ribbed reticulum, giving

a polyhedric appearance of the spores, with 4-5 meshes per spore side and

warts in the inner surface of the meshes. STALK: erect and striate. SPOROTHECA:

globose or subglobose, erect to slightly nodding.

SEM comparisons in Cribaria ... 397

Comments: Although the spores seem to be slightly collapsed, the surface

spore ornamentation is clearly formed by a reticulate pattern with fine warts

inside the meshed areas. ‘These are the first SEM observations of C. dictyospora.

Cribraria intricata Schrad. Nov. Gen. Pl. 7. 1797. PLATE 3e-h

SPECIMEN BPI 828726)—PERIDIUM: partially evanescent, in most of the

sporotheca remaining as a net but sometimes also as a basal disc or cup. NODEs:

thickened, elongated, roughened, connected by <8 slender threads with two or

three free ends. PLASMODIAL GRANULES: concentrated on the outer side of the

nodes. Catycutus: shallow when present, with the upper margin extending

into ribs, which are connected with the peridial net; surface with prominent

radial folds marked by the concentration of the plasmodial granules and fine

concentric folds. When the calyculus is absent, subparallel ribs emerge from

the stalk apex. Spores: globose, 4-5 um diam., with the surface ornamentation

formed by warts; frequently several fused to form an irregular ridge.

COMMENTS: Hatano (1985, 2002) mentions that under SEM the spore surface

of C. intricata has many warts with corrugate swellings. Although in our

specimen most spores were collapsed, our SEM observations appear closer to

those described by Hatano in 1985; our specimen showed a perfectly developed

calyculus, contrasting with the observation by Hatano in 2002 in which the

calyculus was reduced to a basal disc and the ribs emerged from the base of the

sporotheca. As Martin & Alexopoulos (1969) stated, C. intricata is very variable

in these features.

Cribraria languescens Rex,

Proc. Acad. Nat. Sci. Philadelphia 43: 394. 1891 PLATE 4a-c

SPECIMEN TLXM ET44090—PeErip1um: partially fugacious, remaining as

a calyculus at the base and as a net in the upper portion. Nopgs: pulvinate,

isodiametric, connected by 5-6 threads that form an irregular open mesh with

few free ends. PLASMODIAL GRANULES: concentrated on the nodes. CALYCULUs:

shallow, up to half of the sporotheca, with nearly even or somewhat toothed

margins, with faint radial folds. Spores: globose, 6-7 um diam., with the

surface ornamentation formed by remarkably dense, 0.1-0.2 um diam. warts,

several frequently fused to form irregular and sometimes branched ridges.

STALK: long, slender, slightly grooved.

ComMENTs: The surface ornamentation of the spore was described as densely

warted, with the warts forming irregular ridges (Hatano 1985). Our study

corroborates the observations of C. languescens by this author.

398 ... Ramirez-Ortega, Estrada-Torres & De Luna

Cribraria laxa Hagelst. Mycologia 21: 298. 1929. PLaTE 4d-h

SPECIMEN BPI 828894—PERIDIUM: partially evanescent, consisting of ribs

that radiate from the stalk apex, a membranous calyculus at the sporothecal

base, and a very lax net in the upper portion. NoDEs: isodiametric, sometimes

elongated, pulvinate, connected by 4-5 slender threads, with 1-2 free ends,

basal ribs with not connecting threads between them. CALycuLus: irregular,

shallow, formed by thin membrane between the basal ribs, with an irregular

margin. Spores: globose, 5-6 um diam., with the surface ornamentation

formed by 0.3-0.5 um long spines, frequently fused to form irregular ridges in

a sub-reticulate pattern.

ComMMENTs: The spiny ornamentation of the spore in C. laxa forms a

subreticulate pattern, seemingly unique in Cribraria. Although Martin &

Alexopoulos (1969) described a net with elongated and large nodes, we also

frequently observed rounded nodes.

Cribraria lepida Meyl, Bull. Soc. Vaud. Sci. Nat. 56:326. 1927. PLATE 5a-c

SPECIMEN BPI 828797—PERIDIUM: partially evanescent, remaining as a

prominent and deep basal calyculus and an open net at the upper portion of

the sporotheca. Nopgs: irregular, pulvinate, densely covered with plasmodial

granules, connected by 4-5 thin threads, without free ends. CaLycutus: deep,

up to half of the sporotheca, margin even, outer surface granulose due to the

presence of the plasmodial granules. Spores: globose, 6-7 um diam., with the

surface ornamentation formed by warts that frequently fuse to form irregular

short ridges. SPOROTHECA: globose or cylindrical, nodding.

ComMENTs: The spores of C. lepida have been described as smooth using LM

(Martin & Alexopoulos 1969), but they are clearly warted with SEM. Although

this species is similar to C. violacea in color, size, habit, and even the warted

spore ornamentation, the nodes differ considerably: flattened in C. violacea and

pulvinate in C. lepida. As seen with SEM, in C. lepida the nodes resemble a

cluster of grapes due to the aggregation of plasmodial granules.

Cribraria macrocarpa Schrad. Nov. Gen. Pl.: 8. 1797. PLATE 5d-h

SPECIMEN BPI 828967)—PeERIpDIUm: partially fugacious, remaining as a

shallow calyculus at the base and a dense net with very close meshes in the

upper part. NovEs: irregular, large and elongated, flattened, connected by 4-5

wide threads with membranous margins forming a close-meshed net, with

2-3 free ends. PLASMODIAL GRANULES: 2 um diam., smooth, distributed on

SEM comparisons in Cribaria ... 399

the nodes and on the whole calyculus. CALycuLus: occupying one-third the

sporotheca, well developed, margin irregular with many perforations in the

upper portion, with radial folds on outer surface. Spores: globose, 6-7 tm

diam., with the surface ornamentation formed by warts, frequently fused to

form irregular short ridges. STALK: wide and slightly grooved. SPOROTHECA:

globose or ovate, nodding or erect.

ComMENTs: The calyculus with the perforated margin and the flattened and

irregular nodes confirm the identity of C. macrocarpa, based on the LM

description by Martin & Alexopoulos (1969). Adamonyte & Eliasson (2010)

compared it within a species complex comprising C. martinii, C. argillacea, and

C. ferruginea, which all share a perforated peridial calyculus. Nonetheless, we

have no SEM evidence confirming this similarity of C. macrocarpa with the

other species.

Cribraria microcarpa (Schrad.) Pers., Syn. Meth. Fun.:190. 1801. PLATE 6a—d

SPECIMEN BPI 828978—PeERIDIUM: partially fugacious, remaining as ribs

at the base of the sporotheca and a net above; basal ribs not interconnected

with fine threads, emerging from the apex of the stalk. NopEs: small, pulvinate,

hemispherical or sometimes elongated, studded with plasmodial granules,

connected by faint threads that form a close net with polygonal meshes and

without free ends. CALycuLus: absent. Spores: globose, 6-7 um diam., with

the surface ornamentation formed by 0.1 um diam. spines. SPOROTHECA:

globose, erect or nodding. STALK: slender, the apex thinner than the rest of the

stalk, plicate.

CoMMENTs: Hatano (1985) cited two types of spore ornamentation in

C. microcarpa—one with warts and the other with spines. Our observations

agree with Hatano’s second choice, even though our spores were not completely

round. Martin & Alexopoulos (1969) and Hatano (1985) observed that when

present, the calyculus is reduced to a minute basal disc. Our SEM images clearly

show that the basal disc is lacking but that ribs, which radiate from the apex of

the stalk and connect with the peridial net, are present. Nannenga-Bremekamp

(1993) described this feature as “threads springing from the stalk.”

Cribraria mirabilis (Rostaf.) Massee, Monogr. Myxogastr.: 60. 1892. PLATE 6e-h

SPECIMEN TLXM GF44—Peripium: partially evanescent, remaining as

sub-parallel ribs from the base to the middle of the sporotheca, connected by

delicate threads and an irregular net in the upper portion. Noves: flattened,

400 ... Ramirez-Ortega, Estrada-Torres & De Luna

irregular, elongated, connected by 4—5 threads (similar in width to the basal

ribs) with no free ends. PLASMODIAL GRANULES: distributed along the

ribs. CALYCULUs: not observed. Spores: globose, 5-6 um diam., surface

ornamentation composed of winding crests, sometimes branched to form a

subreticulate pattern. SPOROTHECA: globose, erect or nodding.

ComMENts: Cribraria mirabilis has never previously been published

based on SEM observation. SEM observation revealed that the spore

ornamentation is composed of minute crests that sometimes fuse to give

the appearance of ridges. Our SEM images also show that the ribs extend

from the base of the sporotheca to form a well-developed net in the upper

part.

Cribraria oregana H.C. Gilbert, Amer. J. Bot. 19:142. 1932. PLATE 7a-c

SPECIMEN TLXM HC183—PeErIp1vum: partially evanescent, remaining

as a basal shallow calyculus and an irregular net in the upper portion.

Nopes: flattened, irregular, connected by 4-5 threads, which form irregular

meshes and open net with few free ends. CaLycu.us: shallow, with irregular

margins and smooth surface. Spores: globose, 7-8 um diam., with the

surface ornamentation formed by minute and dense warts, frequently fused

to form irregular short ridges. SPOROTHECA: globose, usually erect. STALK:

slender, slightly attenuated at the apex, longitudinally striate.

ComMENTs: This is the first SEM description for C. oregana. Relying on

LM, Martin & Alexopoulos (1969) described the spore ornamentation as

“minutely warted, with a delicate reticulum” while Lado & Pando (1997)

defined it as “spinulose.” Our SEM examination revealed the absence of

both reticulum and spines and the presence of abundant minute warts that

fuse to form irregular short ridges.

Cribraria piriformis Schrad. Nov. Gen. Pl.: 4. 1797. PLATE 7d-h

SPECIMEN BPI 829249—PeERIpIUM: partially fugacious, remaining

as a calyculus at the base of the sporotheca and dense net in the upper

portion. Nopes: isodiametric to elongated, pulvinate, depressed on the

inner side, covered with plasmodial granules, connected by 3-7 diam.

threads that form a closed net and with 3-6 rounded free ends and wider

tips. CALycuLus: shallow, with irregular margins and with some rib-like

projections connecting the peridial net and with pronounced concentric

folds on the outer surface. SPOREs: globose, 6—7 um diam., with the surface

SEM comparisons in Cribaria ... 401

ornamented by abundant minute warts in aggregates. SPOROTHECA:

globose to pyriform, erect. STALK: furrowed, slightly attenuated at the apex.

ComMENTs: Ours is the first SEM description for C. piriformis. It has thickened

nodes as well as wide connecting threads in the peridial net. Nodes on the

inner side are markedly hollow (PI. 7f). Under LM some authors (Martin &

Alexopoulos 1969, Lado & Pando 1997, Adamonyte & Eliasson 2010) have

described the spore ornamentation as densely warted, and our SEM images

confirm this character.

Cribraria purpurea Schrad. Nov. Gen. Pl.: 8. 1797. PLATE 8a-d

SPECIMEN TLXM HC1869—PeEripIum: partially evanescent, remaining

as a shallow basal calyculus and a net in the upper portion. Nopes: flattened,

irregular in size and form, sometimes undifferentiated, connected by 3-6 very

fine threads, with few free ends. PLASMODIAL GRANULES: hollow, on the inner

side of nodes, 2-3 um diam., smooth. Catycutus: shallow, with irregular or

toothed margin, with concentric folds. Spores: globose, 5-6 um diam., with

surface ornamentation composed of winding crests, sometimes branched and

forming a subreticulate pattern. SPOROTHECA: globose, nodding.

ComMENTs: Cribraria purpurea was described using SEM by Moreno et al.

(2001). However, our examination revealed a different spore ornamentation

composed of crests fused to form a sub-reticulate pattern, not observed

in the image provided by Moreno et al. (2001, Fig. 21), who described the

ornamentation as “baculate” (a completely different pattern). It would be

interesting to confirm that both examined specimens represent C. purpurea.

Cribraria rubiginosa Fr. Syst. Mycol. 3:172. 1829. PLATE 8e-h

SPECIMEN TLXM ET2720—PErIp1vu: partially fugacious, remaining as a

deep basal cup and a loose net occupying the upper half of the sporotheca.

NopeEs: irregular, flattened, somewhat angular, filled with plasmodial granules

and connected with strip-like threads, without free ends. PLASMODIAL

GRANULES c. 1-2 um diam. Catycutus: deep, with irregular margins, radial

folds covered with plasmodial granules at least at the base of the sporotheca.

Spores: globose, 6-7 um diam., ornamented by a prominent reticulum formed

by grooves, with four meshes per side, and pronounced warts in the meshes,

sometimes fused. SPOROTHECA: cylindrical, erect. STALK: grooved.

ComMENTs: Martin & Alexopoulos (1969) describe the spores as angular and

with a warted ornamentation by LM. Our SEM image shows a very well defined

402 ... Ramirez-Ortega, Estrada-Torres & De Luna

reticulum formed by grooves that cross the whole spore surface and warts

between the meshes of the reticulum, in a similar way found on the spores

of C. angulospora, a species recently described from Taiwan (Liu & Chang

2007). Nevertheless we suspect that C. angulospora may represent a synonym

of C. rubiginosa. Martin & Alexopoulos (1969) mentioned that the angular

shape of the spore depends on the angle in which the spore is observed in the

microscope, but it is clear that this angularity is due to the presence of the

reticulum.

Cribraria rufa (Roth) Rostaf. Sluzowce monogr.: 232. 1875. PLATE 9a-d

SPECIMEN BPI 829339—PERIDIUM: partially evanescent, remaining as a

shallow basal calyculus and an open-meshed net in the upper portion of the

sporotheca. Nopes: flattened, irregular to elongate connected by 3-5 wide

threads, without free ends. CaLycutus: short, irregular margin, radially plicate

ribs, extending from the base of the sporotheca and connecting with the peridial

net, outer surface with very marked radial folds. SporEs: globose, somewhat

angular, 7-8 um diam., surface ornamentation composed of an open reticulum

formed by thick ribs with warts in the inner areas formed by the meshes of the

reticulum, the warts sometimes fused and forming short ridges. SPOROTHECA:

globose or ovate, erect. STALK: erect, longitudinally striate.

ComMENTs: In a SEM study of Cribrariaceae from Korea, Hatano (2002)

reported the spore ornamentation in C. rufa as a “corrugate swelling [that]

connects each other and makes a vein-like protuberance.” Our SEM observation

of the spore surface shows a lax reticulum divided by spaces with warts included.

Under LM Nannenga-Bremekamp (1991) described the spore ornamentation

as having warts often connected by rows of smaller warts into a lax reticulum.

Our observations agree with the description of spore ornamentation given

by Nannenga-Bremekamp (1991). The angular appearance of the spore, as

mentioned by Martin & Alexopoulos (1969), may be due to the presence of a

lax reticulum on the spore surface.

Cribraria splendens (Schrad.) Pers., Syn. Meth. Fung.:191. 1801. PLATE 9e-h

SPECIMEN TLXM GF1769—PeEripivm: partially fugacious, remaining as

4-5 thick ribs at the base and an open net in upper portion. Nopes: flattened,

not well differentiated, occasionally elongated, without free ends and connected

by 3-4 fine threads, which have membranous margins and form an irregular

and open meshed net; ribs radiating from the stalk and connecting with the

peridial net. CatycuLus: absent. Spores: globose, 5-6 um diam., with the

SEM comparisons in Cribaria ... 403

surface ornamentation formed by warts, sometimes fused to form irregular

short ridges. SPOROTHECA: globose, nodding. STALK: slender, curved at the

apex, longitudinally striate.

COMMENTS: Hatano (2002) described the spore ornamentation for C. splendens

under SEM as “corrugate swellings” that sometimes are fused and then a vein-

like protuberance are observed. Our SEM and his photographs agree, but we

prefer to use the more appropriate term “warts” for this type of ornamentation,

noting that sometimes the warts fuse to form irregular short ridges.

Cribraria tenella Schrad. Nov. Gen. Pl.: 6. 1797. PLATE 10a-d

SPECIMEN TLXM ET4832—PeERIpIvuM: partially fugacious, remaining

in the basal part of the sporotheca as a calyculus, and as a net in the rest.

Nopes: pulvinate, rounded, few free ends, connected usually by four

slender threads. Net irregular, meshes open, with wide connecting threads.

Catycutus: shallow, the margin irregular, surface smooth, sometimes

radial folds emerging from the base, marked by the presence of plasmodial

granules. Spores: globose, minutely warted, the rest of the surface smooth,

5-6 um diam. SPOROTHECA: globose, nodding.

COMMENTS: Hatano (2002) reported the spore surface “with protuberances”

for this C. tenella. In general the peridial net and spore descriptions by

Hatano (1985; 2002) correspond our results, except for the calyculus, which

seem more regular and shallow in Hatano’s description and more irregular

and deeper in our SEM plate.

Cribraria violacea Rex, Proc. Acad. Nat. Sci. Philadelphia 43:393. 1891. PLATE 10e-h

SPECIMEN BPI 829575—PERIDIUM: partially evanescent, remaining

as a deep calyculus at the base and as a net in the upper portion. NODEs:

flattened, irregular in size and form, with hollow protuberances due to the

plasmodial granules, connected by delicate threads, which form an open

net, with few and irregular meshes, without free ends. CALycULUs: deep,

occupying half or two thirds of the sporotheca, margin toothed, with the

surface studded with plasmodial granules. Spores: globose, 6—7 um diam.,

with the surface ornamentation formed by warts, sometimes fused forming

irregular short crests. SPOROTHECA: globose or cylindrical, erect. STALK:

slender, grooved.

ComMENTs: Moreno et al. (2001) are the only authors to have described

this species as “baculate with small crests” using SEM. The two SEM images

404 ... Ramirez-Ortega, Estrada-Torres & De Luna

are similar, but our SEM plate shows warts that are much closer and form

irregular short crests. Under LM most authors have described C. violacea as

minutely warted (Martin & Alexopoulos 1969, Nannenga-Bremekamp 1991,

Lado & Pando 1997).

Cribraria vulgaris Schrad. Nov. Gen. Pl.: 6. 1797. PLATE 11

SPECIMEN BPI 829689—PeERIpIUM: partially fugacious, remaining as

a calyculus at the base and as a net in the upper portion of the sporotheca.

Nopes: irregular or isodiametric, flattened, connected by 4-5 threads that

form an irregular net without free ends. CALYCULUS: occupying about one

third of the lower portion of the sporotheca, with irregular margins, forming

spiny teeth that give rise to the net, the surface marked by delicate radial folds.

SPORES: 5-6 um diam., the surface ornamented by warts that sometimes fuse

to form irregular short ridges (not possible to confirm in collapsed spores).

SPOROTHECA: globose, nodding. STALK: erect, slightly attenuate at the apex,

longitudinally striate.

COMMENTS: Martin & Alexopoulos (1969) described the spore

ornamentation of C. vulgaris based on LM as “minutely punctate;”

nevertheless Nannenga-Bremekamp (1991), Lado & Pando (1997), and

Adamonyte & Eliasson (2010) described it as “slightly angular, warted with

faint ridges or with wartlets forming a lax reticulum.” Warts are present and

distributed on the spore surface in our SEM photographs, but the presumed

reticulum was not observed due to the collapsed spore.

Character coding under phylogenetic perspective

Character description plays such an essential role in the phylogenetic

inference process, determination of characters and character states is one of

the most fundamental steps in any taxonomic work (Kirchoff 2001). Newton

& De Luna (1999) noted, “we need not ascertain a priori if a character or its

states have a common origin, or if character states are homologous” because

these result from the cladistics analysis. The final result from the inference

cladistics process must serve as the foundation for any discussion about

evolutionary character change that will allow us to corroborate or change

the homology hypothesis based on strict cladistic principles (Wirkner &

Richter 2010).

In a phylogenetic context, Sereno (2007) defined the logical basis

for morphological characters, where he concluded that “characters are

simply features expressed as independent variables and character states

SEM comparisons in Cribaria ... 405

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a :

PLATE 1. Cribraria atrofusca. a. Sporocarp; b. Enlarged node with plasmodic granules; c. Margin

and outer surface of calyculus; d. Collapsed spore. Cribraria aurantiaca. e. Sporocarp; f. Node;

g. Margin and outer surface of calyculus; h. Surface spore ornamentation. Scale bars: a, e = 100 um;

b= 10 um; c= 20 um; d= 2 um; f=5 wm; g=50 um; h=1 pm.

406 ... Ramirez-Ortega, Estrada-Torres & De Luna

PLATE 2. Cribraria cancellata. a. Sporocarp; b. Peridial ribs emerging from the stalk apex; c. Spore

and plasmodic granule. Cribraria costata. d. Sporocarp; e. Node of peridial net; f. Peridial net

without free ends; g. Spore surface ornamentation with crestae; h. Spores. Scale bars: a = 100 um;

b, f = 20 um; c, g = 1 um; d = 50 um; e =10 um; h = 5 um.

SEM comparisons in Cribaria ... 407

he h

PLATE 3. Cribraria dictyospora. a. Margin details and outer surface of calyculus; b. Node;

c. Collapsed spore with reticulum; d. Collapsed spores showing surface reticula. Cribraria

intricata. e. Sporocarp; f. Peridial node with 2-3 free ends; g. Calyculus margin connecting with

the peridial net and details of the outer surface; h. Spore surface ornamentation with warts. Scale

bars: a = 50 um; b, f = 10 um; c= 2 um; d = 5 um; e = 100 um; g = 20 um; h = 1 um.

408 ... Ramirez-Ortega, Estrada-Torres & De Luna

PLATE 4. Cribraria languescens. a. Sporocarp; b. Peridial node apparently without free ends;

c. Spore surface ornamentation with warts. Cribraria laxa. d. Peridial net; e. Peridial ribs emerging

from the base of the sporocarp and details of the calyculus surface; 50 um. f. Thickened node;

g. Spores; h. Spore ornamentation. Scale bars: a, e = 50 um; b = 5 um; c, h = 1 um; d = 100 um;

f= 10 um; g = 2 um.

SEM comparisons in Cribaria ... 409

PLaTE 5. Cribraria lepida. a. Sporocarp; b. Peridial node; c. Spore. Cribraria macrocarpa.

d. Sporocarp showing the peridial net and calyculus; e. Peridial node with plasmodic granules

and spores; f. Details of calyculus outer surface; g. Spore surface ornamentation; h. Spores and

plasmodic granules. Scale bar: a = 20 um; b = 5 um; c, g = 1 um; d = 100 um; e = 10 um; f = 50 um;

h=2 um.

410 ... Ramirez-Ortega, Estrada-Torres & De Luna

— he ans

PLaTE 6. Cribraria microcarpa. a. Peridial net of the sporocarp; b. Peridial nodes with plasmodic

granules; c. Peridial ribs emerging from the apex of the stalk; d. Spore with spines as surface

ornamentation. Cribraria mirabilis. e. Sporocarp showing the peridial ribs and net; Peridial

net without well-defined nodes; g. Spore surface ornamentation; h. Spore. Scale bars: a = 50 um;

b, c, f= 10 um; d, g, h = 1 um; e = 200 um.

SEM comparisons in Cribaria ... 411

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PLaTE 7. Cribraria oregana. a. Sporocarp; b. Spore surface ornamentation; c. Spore with warts.

Cribraria piriformis. d. Peridial net and spores; e. Outer surface of calyculus and margin

connecting with the peridial net; f. Peridial node with 2-3 free ends; g. Spores; h. Spores and

plasmodic granules. Scale bars: a, d = 50 um; b, c, g,h = 1 um; e = 100 um; f = 20 um.

412 ... Ramirez-Ortega, Estrada-Torres & De Luna

PLATE 8. Cribraria purpurea. a. Peridial net; b. Nodes flattened without free ends; c. Spores with

surface ornamentation composed of crests; d. Spores and plasmodic granules. Cribraria rubiginosa.

e. Flattened node with spores and plasmodic granules; f. Inner surface details of calyculus; g. Spore

details of surface ornamentation showing warts and a reticulum groove crossing the spore; h. Spore

seen in angular view. Scale bars: a =100 um; b = 20 um; c, d= 2 um; e, f= 10 um; g, h = 1 um.

SEM comparisons in Cribaria ... 413

PLatE 9. Cribraria rufa. a. Sporocarp; b. Peridial ribs extending from the base of the sporotheca;

c. Spore showing a reticulum and warts in the inner area; d. Spores. Cribraria splendens.

e. Sporocarp; f. Node flattened and elongated; g. Peridial ribs extending from the apex of the stalk;

h. Spore surface ornamentation with warts. Scale bars: a, e =100 um; b = 50 um; c, h = 1 um;

d=2 um; f, g = 20 um.

414 ... Ramirez-Ortega, Estrada-Torres & De Luna

os fe Ne:

PiatE 10. Cribraria tenella. a. Sporocarp; b. Pulvinate and rounded node; c. Calyculus with radial

folds emerging from the base; d. Spore surface ornamentation with warts. Cribraria violacea.

e. Sporocarp; f. Peridial node with hollow protuberances caused by plasmodic granules; g. Calyculus

surface (detail); h. Spore with surface ornamentation formed by warts. Scale bars: a, e =50 um;

b,h=2 um; c, fg = 10 ym; d = 1 pm.

SEM comparisons in Cribaria ... 415

PLATE 11. Cribraria vulgaris. a. Sporocarp; b. Peridial node flattened and without free ends;

c. Outer surface of calyculus and margin connecting with the peridial net; d. Collapsed spore with

warts as spore surface ornamentation. Scale bars: a = 100 um; b = 5 um; c = 50 um; d= 1 um.

the mutually exclusive conditions of that character.’ Four components for

character statements were recognized: locators, variable, variable qualifiers,

and characters states. All of this information can be expressed in symbolic

form (for more details see Sereno 2007: Table 4).

The rationale for defining character and character states (character

conceptualization), therefore, is a crucial step in evolutionary morphology.

In order to render the information obtained with the use of SEM for the

genus Cribraria, we have applied the scheme proposed by Sereno (2007) for

morphological characters with a symbolic connotation. Here we summarize

the data obtained for the specimens analyzed whose phylogenetic

information could be tested through a cladistics framework. These results

were interpreted as suggesting that a character has at least two different

character states (variables feature), and considering all the range variation

observed for the species included in the present study (see TABLE 1, p. 416).

416 ... Ramirez-Ortega, Estrada-Torres & De Luna

TABLE 1. Symbolic notation for characters and character states in sporotheca and

spores of the Cribraria species.

CHARACTER (LOCATOR) CHARACTER STATES (VARIABLES) SYMBOLIC NOTATION*

SPOROTHECA

Peridial net (0) Absent; (1) present Lz: ie ;V

Peridial ribs (0) Absent; (1) present L: ee : ve

Calyculus (0) Absent; (1) present Teg Vi ;V

Nodes differentiated (0) Absent; (1) present L: vA ; Ve

Thickness of nodes (0) Pulvinate; (1) flattened L: vi, ;V

Type of nodes in (0) Enlarged; (1) isodiametric;

peridium (2) irregular LL Vo VV,

Calyculus upper margin (0) Entire; (1) irregular; (2) ribbed; (3) Toothed L, Li Vig Sas V3 V;

Calyculus surface (0) Radial folds; (1) concentric folds LL: V;3V

SPORE SURFACE ORNAMENTATION

Warts (0) Absent; (1) present L: Ne ; xe

Crests (0) Absent; (1) present Lis Vy ; Vv,

Reticulum (0) Absent; (1) present Le Vy ; We

Bands (0) Absent; (1) present L Q vs : ¥,

Spiny-subreticulate (0) Absent; (1) present L: Vi, : Va

Grooved-reticulate (0) Absent; (1) present Lj: Vo : Ne

*L, = primary locator, L, = secondary locator; V_ = character state

Conclusions

In this paper, we have described spore ornamentation variables (character

states) as revealed by SEM never before noted for species such as C. ferruginea

and C. laxa. Studies of micro-morphological characters in Cribraria with

SEM, such as spore ornamentation and the shape of nodes, demonstrate that

interpretations of these character states based on LM alone may be incorrect

or completely lacking. This is because LM does not make certain features

prominent or even noticeable in the same way as SEM. Cribraria, as observed

by Ing (1999) and Adamonyte & Eliasson (2010), is a challenging genus due

to overlapping morphological variation among the various species, making

it problematical to circumscribe particular features with accuracy. Therefore,

SEM is a fundamental tool for taxonomic studies in this genus as well as for

other myxomycete taxa for which similar empirical difficulties exist. Further

SEM research in Cribraria, either of same species or those not included in this

work, will certainly confirm or amplify the morphological variation noted here.

SEM comparisons in Cribaria ... 417

Acknowledgments

We are deeply grateful to Dr. Steve Stephenson (University of Arkansas) and Dr.

Yuri Novozhilov (Komarov Botanical Institute) for their critical pre-submission peer

review with comments and suggestions to improve this manuscript. We also appreciate

the valuable helpful comments and detailed preliminary review by Dr. Harold Keller

(Botanical Research Institute of Texas). Thanks a lot to Dr. Stephenson for his valuable

linguistic assistance. We especially thank Dr. Amy Rossman (USDA) for the loan of

myxomycete collection from BPI and Dr. Hagiwara (National Museum of Nature and

Science, Tsukuba, TNS) for donation of biological material. Funding for this work was

provided by the Consejo Nacional de Ciencia y Tecnologia de México (CONACYyT)

under the CONACyT-SEP 2003-C02-44621 project.

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SEM comparisons in Cribaria ... 419

Glossary

Basal disc: A small portion of the calyculus remaining at the base of the

sporotheca.

Calyculus: The lower portion of the sporotheca remaining as a persistent

structure like a cup (Keller & Braun 1999)

Nodes: “The thickened area at the point of juncture where several threads meet

to form the peridial network in the upper part of the sporotheca” (Keller &

Braun 1999).

Peridial net: A persistent peridium that remains as a reticulate structure in the

sporotheca (from Martin & Alexopoulos 1969).

Peridial ribs: Subparallel elements of the peridium, interconnected by short

transverse threads, which may arise from the base of the sporotheca or

extend from the calyculus margin.

Plasmodial granules: Microscopic, usually dark-colored structures found in

the sporotheca of Cribraria, on either the peridial nodes or calyculus. These

have been called “dictydine granules” (Martin & Alexopoulos 1969; Keller &

Braun 1999) or “lime globules” (Nannenga-Bremekamp 1991). We prefer to

use the term “plasmodial granules” based on their plasmodial origin.

Spore ornamentation: The surface of the spore, as seen under light microscope,

may be smooth or have variously formed projections. Ornamentation

features include:

Bands: Flat (generally wide) regions that typically cross to form a reticulate

pattern.

Crests: projections distributed in waves.

Grooved-reticulate: a net-like pattern (reticulum) formed by deep grooves

or furrows.

Reticulum: the pattern produced by warts distributed in lines that cross in

a net-wise fashion.

Warts: short obtuse projections distributed over the surface.

MY COTAXON

April-June 2017— Volume 132, pp. 421-432

https://doi.org/10.5248/132.421

New records of Ascomycota

from tropical dry forest in Sonora, Mexico

TANIA RAYMUNDO ', MARTHA L. CORONADO ’,

ALDO GUTIERREZ ?, MARTIN ESQUEDA 3? & RICARDO VALENZUELA ‘**

' Escuela Nacional de Ciencias Bioldgicas, Instituto Politécnico Nacional,

Plan de Ayala y Carpio s/n Col. Santo Tomas, México, D.F. 11340, México

? Universidad Estatal de Sonora,

Apartado Postal 11, Admon. 11, Hermosillo 83000, Sonora, México

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

Km 0.6 Carretera a La Victoria, Hermosillo, Sonora 83304, México

* CORRESPONDENCE TO: rvalenzg@ipn.mx

ApsTract—Eight new records of ascomycetes from Sierra de Alamos-Rio Cuchujaqui

Biosphere Reserve in Sonora, Mexico are reported. Hypoxylon anomalum and Munkovalsaria

donacina represent new records for Mexico, while Bionectria byssicola, Fracchiaea

heterogenea, Hypoxylon lenormandii, Lasiodiplodia theobromae, Peroneutypa scoparia, and

Pseudocosmospora vilior are new records for Sonora.

Key worps—Botryosphaeriales, Coronophorales, Hypocreales, Xylariales

Introduction

The Sierra de Alamos-Rio Cuchujaqui Biosphere Reserve (ACBR)

presents an extraordinary biological richness, with more than 1200 plant

species in an area of 92,899 ha. It is located in the south of Sonora, Mexico,

preserving part of the North American tropical dry forest intermixed with

xerophilous scrub, oak-pine forest, and riparian vegetation (GSEMARNAT

2015). Its floral and faunal components have been studied by Robichaux &

Yetman (2000).

Twenty-two ascomycetes been recorded previously from ACBR:

Anteaglonium abbreviatum (Schwein.) Mugambi & Huhndorf [= Glonium

abbreviatum (Schwein.) M.L. Lohman], Biscogniauxia nummularia (Bull.)

422 ... Raymundo &al.

Kuntze, Daldinia clavata Henn., D. concentrica (Bolton) Ces. & De Not.,

D. eschscholtzii (Ehrenb.) Rehm, H. ferrugineum G.H. Otth [= H. rubiginosum

var. ferrugineum (G.H. Otth) J.H. Mill.], D. fissa Lloyd, D. loculata (Lév.)

Sacc., Hypoxylon anthochroum Berk. & Broome, H. haematostroma Mont.,

H. polyporus (Starback) Y.M. Ju & J.D. Rogers, H. rubiginosum (Pers.) Fr.,

Hysterobrevium mori(Schwein.) E. Boehm & C.L. Schoch [=Hysterographium

mori (Schwein.) Rehm], Oedohysterium insidens (Schwein.) E. Boehm &

C.L. Schoch, Rhytidhysteron rufulum (Spreng.) Speg., Xylaria arbuscula

Sacc., X. feejeensis (Berk.) Fr., X. microceras (Mont.) Berk., X. poitei

(Lév.) Fr., X. striata Pat., X. striata var. sonorensis F. San Martin et al., and

X. uniapiculata FE. San Martin & J.D. Rogers (Pérez-Silva et al. 1996; San

Martin et al. 1999a,b; Méndez-Mayboca et al. 2010; Chacon et al. 2014).

This study aims to continue the expansion of our knowledge of ascomycete

diversity in the ACBR.

Materials & methods

The Sierra de Alamos-Rio Cuchujaqui Biosphere Reserve was surveyed with two

samplings in October of 2013 and 2014, in the localities of El Cajon (27°01’42”N

108°45’23”W, 373 m a.s.l.) and Rancho La Sierrita (26°58’25”N 108°56’42”W, 528

m a.s.l.). The collected specimens were described from fresh material, processed,

and deposited in the Herbarium, Escuela Nacional de Ciencias Bioldgicas, Instituto

Politécnico Nacional, Mexico City, Mexico (ENCB). The species were determined

by observing cut sections of the ascomata in temporary preparations mounted in

70% alcohol, 10% KOH, and lactophenol with cotton blue (LB) and compared with

identification keys (Aptroot 1995a,b; Ju et al. 2005; Burgess et al. 2006; Carmaran et al.

2006; Chaverri et al. 2011; Herrera et al. 2013). New records for Sonora are indicated by

a single asterisk, and for Mexico with two asterisks.

Taxonomy

Dothideomycetes, Botryosphaeriales, Botryosphaeriaceae

*Lasiodiplodia theobromae (Pat.) Griffon & Maubl.,

Bull. Soc. Mycol. France 25: 57 (1909) Figs 1, 2

Pycnipi of (360-)470-510(-540) x (210—)350-430(-500) um, subglobose,

obpyriform to pyriform, dark brown to black, immersed in the surface of

the host, erumpent when mature. PARAPHYSES hyaline, cylindrical, septate,

occasionally branched, with rounded tips of 45-55 x 3-4 um. Conidiogenous

cells, cylindrical, hyaline, thin and smooth wall. Conrpia of (19-)21-31(-32.5)

x (12-)13-15.5(-18.5) um, subovoid to ovoid-ellipsoid, broadly rounded apices

tapering or truncate at the base, initially hyaline and aseptate with a thick wall,

New ascomycetes for Sierra de Alamos-Rio Cuchajaqui (Mexico) ... 423

dark olive, dark brown to black, with a transverse septum and longitudinal

grooves or sulci when mature.

SPECIMENS EXAMINED: MEXICO, Sonora: El Cajon, 14 Oct 2013, T. Raymundo 4819,

4827; Rancho La Sierrita, 15 Oct 2013, T. Raymundo 4827, D. Castro-Bustos 164.

HABITAT & DISTRIBUTION—Widely distributed in tropical and subtropical

regions, L. theobromae grows gregariously on decaying fruit such as Ceiba

acuminata (S. Watson) Rose, and Alves et al. (2008) indicate that it is plurivorous.

In México, it has been reported from the states of Campeche, Colima, Distrito

Federal, Estado de México, Guerrero, Guanajuato, Jalisco, Michoacan, Nayarit,

Puebla, Querétaro, Tabasco, Tlaxcala, and Veracruz (Medel & Chacén 1988,

Cibrian- Tovar et al. 2007). This is a first record for Sonora.

ComMENTS—Lasiodiplodia theobromae is recognized by its black pycnidia,

immersed and erumpent when mature, which produce subovoid and dark

brown conidia with conspicuous longitudinal grooves. It is considered

a virulent plant pathogen of hundreds of host species around the world. In

Mexico, Cibrian-Tovar et al. (2007) consider it an opportunistic fungus that

attacks stressed trees and produces various toxins that could favor leaf wilt in

several tropical trees. A similar species, L. pseudotheobromae A.J.L. Phillips et

al., differs by its dark brown and slightly larger (23.5-32 x 14-8 um) conidia

and occurrence in Costa Rica on Acacia mangium, Citrus aurantium, Coffea

sp., and Rosa sp. (Alves et al. 2008). Lasiodiplodia parva A.J.L. Phillips et al. is

also similar but produces smaller (16-23 x 10-13 tm) conidia and has been

recorded only on Theobroma cacao in Colombia and Sri Lanka (Alves et al.

2008).

Dothideomycetes, inc. sed., Dacampiaceae

**Munkovalsaria donacina (Niessl) Aptroot,

Nova Hedwigia 60: 346 (1995) Fics 3,4

STROMA inconspicuous, pseudothecia immersed, erumpent; solitary to

gregarious, globose, (125-)200-250 um diam., with a conspicuous ostiole.

PERIDIUM a textura angularis, 10-15 um thick, with 3-6 hyaline layers on

the inner side, golden brown on the outside; pseudoparenchyma formed by

subglobose to angular cells 5-6.5 um diam. HAMATHECIA with numerous

pseudoparaphyses, 1-2.5 um diam., with septa spaced at intervals of 10-20

um, anastomosing, hyaline, inamyloid. Asci bitunicate, hyaline when young,

50-70 x 6-9 um, cylindrical-clavate, with long stipes, octosporous, obliquely

uniseriate to biseriate apically when mature. Ascospores 14-17 x 6-7.6 um,

424 ... Raymundo &al.

ellipsoidal to clavate, asymmetrical with a central septum that is transverse,

constricted, and slightly submedian, brown yellowish and reddish brown with

two green lipid droplets when mature, with slightly sharp ends; wall thin,

slightly spinose, without gelatinous cover.

SPECIMEN EXAMINED: MEXICO, Sonora: El Cajon, 14 Oct 2013, E. Escudero-Leyva

244b.

HABITAT & DISTRIBUTION—Cosmopolitan and pantropical species. Gregarious,

immersed in wood of woody angiosperms, where it coexists with Peroneutypa

scoparia. Munkovalsaria donacina has been reported in a variety of hosts, mainly

Acanthaceae, Anacardiaceae, Fabaceae, Malvaceae, Moraceae, Juglandaceae,

Rutaceae, and Verbenaceae. Reported from Africa, Asia, Australasia, Europe,

North America, and South America (Aptroot 1995b). This is the first record

from México.

ComMENtTs—Munkovalsaria donacina is characterized by gregarious

pseudothecia. A similar species, M. appendiculata Aptroot is distinguished by

its ascospores (12-15 x 4-5 um, fusiform with two hyaline polar appendages),

growth on Zea mays in secondary vegetation, and distribution in Hong Kong

(Aptroot 2004). Didymosphaeria conoidea Niessl is also similar but produces

ascospores that are 9-12 x 4-6 um, ellipsoidal to clavate, and with slightly sharp

ends. Munkovalsaria is characterized by hamathecia with transversely septate

pseudoparaphyses and euseptate ascospores, while Didymosphaeria produces

hamathecia with narrow trabeculate paraphysoids and thinly distoseptate

ascospores (Aptroot 1995b).

Sordariomycetes, Coronophorales, Nitschkiaceae

*Fracchiaea heterogenea Sacc.,

Atti. Soc. Veneto-Trentino Sci. Nat. 2: 163 (1873) Fics 5, 6

PERITHECIA gregarious in groups of 20-40, rarely solitary, erumpent, sessile,

subglobose to globose, 500-700 um diam., completely closed without ostiole,

non-collapsing when dry, glossy black. Surface when young black verrucose with

18-25 um long spines, becoming smooth when mature. Wall a textura angularis,

with thick-walled pseudoparenchymatous cells with Munk pores. Quellk6érper

rudimentary. PARAPHYSES absent. Ascr 80-100 x 15-18 um, cylindrical,

rounded at the apical end, with short stipes, multispored (<200 ascospores),

multiseriate, arranged obliquely in several longitudinal rows. AscosPporREs

hyaline, 7.6-9 x 1.5-2 um, cylindrical, slightly curved to allantoid, eventually

with central septum and lipid droplets at the ends, walls thin and smooth.

New ascomycetes for Sierra de Alamos-Rio Cuchajaqui (Mexico) ... 425

Fics 1-8. Lasiodiplodia theobromae. 1. Pycnidia. 2. Conidia. Munkovalsaria donacina.

3. Pseudothecia. 4. Ascus and ascospores. Fracchiaea heterogenea. 5. Perithecia in the substrate.

6. Perithecium in KOH. Bionectria byssicola. 7. Perithecia in the substrate. 8. Perithecium in KOH.

SPECIMENS EXAMINED: MEXICO, Sonora: Rancho La Sierrita, 15 Oct 2013, T.

Raymundo 4848, R. Valenzuela 15262, D. Castro-Bustos 156.

HABITAT & DISTRIBUTION—Cosmopolitan according to Nannfeldt (1975).

Fracchiaea heterogenea grows on decaying branches of newly fallen trees,

and is considered saprophytic and/or parasitic on other fungi. Chaverri et

426 ... Raymundo & al.

al. (2011) recorded it for Costa Rica. In Mexico, it has been reported from

Tamaulipas and Oaxaca (San Martin 1996, Raymundo et al. 2014). First

record for Sonora.

COoMMENTS— The studied material matches the description by San Martin

(1996), who reported F. heterogenea for the first time from Mexico in El Cielo

Biosphere Reserve. Nannfeldt (1975) and San Martin (1996) considered it

synonymous with Nitschkia broomeana (Berk.) Nannf., but Nitschkia species

have octosporous asci, while Fracchiaea species have multisporous asci.

We follow these criteria. The anamorph is not known yet.

Sordariomycetes, Hypocreales, Bionectriaceae

*Bionectria byssicola (Berk. & Broome) Schroers & Samuels,

Z. Mykol. 63: 152 (1997) Fics 7, 8

PERITHECIA 290-400 um diam., globose to subglobose, orange to pink

when fresh, pale yellowish and laterally depressed when mature; no color

change in KOH. Wall with whitish verrucae, 25-35 um thick, dark red, textura

epidermoidea, cells 10.4-12.8 x 8-12.8 um. Ascr 65-70 x 6-8.4 um, clavate,

with a visible apical ring in LB, hyaline, octosporous, uniseriate. ASCOSPORES

13.6-15.2 x 4.8-5.6 um, ellipsoidal to oblong, with a septum in the middle,

hyaline, thin-walled, finely spinulose.

SPECIMEN EXAMINED: MEXICO, Sonora: Rancho La Sierrita, 15 Oct 2013, R.

Valenzuela 15272.

HABITAT & DISTRIBUTION—Bionectria byssicola, which grows gregariously on

trunks of newly fallen trees, has been cited from China, Taiwan, and Costa

Rica (Nong & Zhuang 2005, Guu et al. 2010, Chaverri et al. 2011). In México

it has been reported from the National Park Lagunas de Chacahua, Oaxaca

(Raymundo et al. 2014). First record for the Sonoran mycobiota.

CoMMENTS—Bionectria byssicola is characterized by its orange to pink

perithecia with whitish verrucae that change to pale yellowish in KOH. The

anamorph [described as Clonostachys byssicola Schroers] forms sporodochia

with curved conidia. Our Sonoran material matches the descriptions of

Chinese and Taiwanese material (Chaverri et al. 2011, Guu et al. 2010). A

similar species is B. grammicospora (Ferd. & Winge) Schroers & Samuels,

which differs by its orange perithecia that do not change color in KOH,

ascospores of 12.8-13.6 x 4.4-4.8 um, and by the synnema in its anamorph

[described as Clonostachys grammicospora Schroers & Samuels] (Schroers &

Samuels 1997).

New ascomycetes for Sierra de Alamos-Rio Cuchajaqui (Mexico) ... 427

Sordariomycetes, Hypocreales, Nectriaceae

*Pseudocosmospora vilior (Starback) C. Herrera & P. Chaverri,

Mycologia 105: 1301 (2013) Fics 9, 10

PERITHECIA 209-248 x 171-199.5 um, obpyriform, red changing to dark

red in KOH, smooth, inconspicuous stroma. Neck 95-110.2 x 28.5-43.7

um, occasional periphyses emerging from the ostiole (inconspicuous to

the naked eye). Wall smooth, 20-40 um thick, orange or yellowish in LB,

textura epidermoidea; cells 8-12 x 2.4-4.8 um. Asci 50.4-57.6 x 5.4-6.4 um,

cylindrical, without apparent apical apparatus, hyaline, octosporous, uniseriate.

ASCOSPORES 6.4-8.4 x 4-5.6 um, ellipsoidal, constrained by a central septum,

hyaline to yellowish, thin-walled, verrucose.

SPECIMEN EXAMINED: MEXICO, Sonora: El Cajon, 14 Oct 2013, T. Raymundo 4808.

HABITAT & DISTRIBUTION—Pseudocosmospora vilior grows gregariously on the

surface of Xylariaceae. It has been recorded from China, Taiwan, and Costa

Rica (Nong & Zhuang 2005, Guu et al. 2007, Chaverri et al. 2011). In México

it has been reported from the National Park Lagunas de Chacahua, Oaxaca

(Raymundo et al. 2014). It is cited for the first time for Sonora.

CoMMENTS—Pseudocosmospora vilior is characterized by smooth red perithecia

that change to dark red with a white ostiole in KOH. Our Sonoran material

matches the descriptions of Costa Rican and Taiwanese material (Chaverri et

al. 2011, Guu et al. 2007). Cosmospora flammea (Tul. & C. Tul.) Rossman &

Samuels is similar, but produces perithecia that emerge in dense clusters from

wood or scale insects and ascospores of 16-20 x 7.5-10 um (Herrera et al.

2013). Macroconia gigas (J. Luo & W.Y. Zhuang) Grafenhan & Seifert is also

similar, differing by its perithecia with yellowish trichomes and larger (13.5-23

x 5.5-7 um) ascospores (Luo & Zhuang 2007).

Sordariomycetes, Xylariales, Diatrypaceae

*Peroneutypa scoparia (Schwein.) Carmaran & A.I. Romero,

Fungal Diversity 23: 84 (2006) Fics 11, 12

= Peroneutypa heteracantha (Sacc.) Berl., Icon. Fung. 3: 81 (1902)

STROMATA 3-5 mm diam., pustuliform, black, erumpent, semicircular,

with small bulges formed by 3-13 perithecia per stroma, monostichous or

polystichous. PERITHECIA 400-600 um diam., globose to ovoid, with thick

black wall and surrounded by xylem tissue. Necks 800-1200 x 80-120 um,

extending conspicuously above the stroma, with cruciform ostiolar openings,

surrounded by a velvety layer of yellowish to pale brown mycelia. Asci 10-18

428 ... Raymundo & al.

x 4-5 um, clavate, octosporous with amyloid apical ring. AscospoREs 4-5 x

1-1.5 um, pale yellow in KOH, greenish in mass, moderately allantoid, with

two lipid droplets at the ends, aggregated to biseriate in the ascus.

SPECIMEN EXAMINED: MEXICO, Sonora: Rancho La Sierrita, 15 Oct 2013, T.

Raymundo 4857.

HABITAT & DISTRIBUTION—This species grows gregariously on decaying

debarked wood in tropical dry forests. Reported from Africa, Europe, Asia,

North America, and South America (Rappaz 1987). In Mexico, P. scoparia

has been reported from México City, Morelos, Nuevo Leén, Oaxaca, San Luis

Potosi, and Tamaulipas (Pérez-Silva 1986, as P. heteracantha; Chacon 2003;

Carmaran et al. 2006; Raymundo et al. 2014). This is the first record for Sonora.

CoMMENTS—Peroneutypa scoparia is characterized by pustuliform erumpent

stromata and by perithecia with prominent and cruciform ostiolar necks at

the apex. Our Sonoran material matches the description of Rappaz (1987). A

similar species is Eutypella portoricensis (Petr.) Rappaz, with similar stromata,

prominent but thin ostiolar necks, and larger (4-7 x 1.5-1.8 um) ascospores

(Rappaz 1987).

Sordariomycetes, Xylariales, Xylariaceae

**Hypoxylon anomalum J.D. Rogers, Y.M. Ju & Oses,

Mycologia 97: 562 (2005) Fics 13, 14

STROMA 4-6 mm diam., aggregated, with grouped perithecia, conspicuous.

The tissue between the perithecia is not to slightly conspicuous, pale orange.

PERITHECIA 400-600 um diam., obovoid, orange to ocher brown and glossy

dark brown, releasing pale yellowish pigments in KOH. Ostioles with umbilicate

surface. Asc1 80-98 x 10-12 um, stipes 40-66 um long, apical ring 1.4-2 x

3-4 um, discoid, inamyloid, octosporous, uniseriate. AscospoREs 12-15.5 x

6.5-8 um, inequilaterally ellipsoid, with broadly rounded ends, dark brown to

black; rectilinear germinal aperture with dehiscent perisporium and smooth

episporium.

SPECIMENS EXAMINED: MEXICO, Sonora: El Cajon, 14 Oct 2013, T. Raymundo 4809;

Rancho La Sierrita, 15 Oct 2013, R. Valenzuela 15249.

HABITAT & DISTRIBUTION—Hypoxylon anomalum grows gregariously on

decaying branches. Reported in Costa Rica (Ju et al. 2005); it is recorded for the

first time for México.

CoMMENTS—Our Mexican material matches the description of H. anomalum

by Ju et al. (2005). A similar species, Hypoxylon olliforme San Martin et al., is

New ascomycetes for Sierra de Alamos-Rio Cuchajaqui (Mexico) ... 429

Fics 9-16. Pseudocosmospora vilior. 9. Perithecia in Hypoxylon. 10. Perithecium in KOH.

Peroneutypa scoparia. 11. Stromata and perithecia. 12. Asciand ascospores. Hypoxylon anomalum.

13. Stromata and perithecia. 14. Ascospores. Hypoxylon lenormandii. 15. Stroma and perithecia.

16. Ascus and ascospores.

430 ... Raymundo & al.

distinguished by its stroma that is effuse, pulvinate to rosellinoid, reddish

brown and releasing reddish orange pigments in KOH, globose (200-300

um diam.) perithecia, and ascospores that are 10.5-12 x 4.5-5 um, dark

brown, ellipsoidal (San Martin et al. 1999d).

*Hypoxylon lenormandii Berk. & M.A. Curtis,

J. Linn. Soc., Bot. 10: 385 (1868 [“1869”]) Fics 15, 16

STROMA 1500-10000 x 800-4000 x 1.5-2 mm, flat to effuse-pulvinate;

perithecia not to slightly conspicuous, surface violet to violet brown with

dark granules, sepia when mature, with removable yellowish green to olive

pigments. The tissue between the perithecia is inconspicuous to slightly

conspicuous, black, 200-700 um thick. PERITHECIA 500-1000 x 300-400

um, tubular, black, with ostioles below the stromal surface opening the

stromal layer. Asc1 72-98 x 4.5—6 um, with 28-56 um long stipes, apical ring

of 1.4-2 x 3-4 um, discoid, amyloid, octosporous, uniseriate. ASCOSPORES

7-9 (-12) x 3.5-5.5 um, ellipsoidal with broadly rounded ends, dark brown,

rectilinear germinal aperture with indehiscent perisporium and smooth

episporium.

SPECIMEN EXAMINED: MEXICO, Sonora: El Cajon, 14 Oct 2013, D. Castro-Bustos

129, 144,

HABITAT & DISTRIBUTION—Hypoxylon lenormandii grows gregariously on

decaying branches and trunks from fallen trees. Recorded in United States

(Vasilyeva et al. 2007) and México, where it has been reported in Campeche,

Chiapas, Nuevo Leon, Oaxaca, Quintana Roo, and Tamaulipas (San Martin

et al. 1999c; Raymundo et al. 2014). This is the first record for Sonora.

CoMMENTsS—Our Sonoran material of H. lenormandii matches the

description by San Martin et al. (1999c) for Mexican specimens; it is mostly

consistent with the United States specimens, which have somewhat larger

(10-15 x 4-6.5 um) ascospores (Vasilyeva et al. 2007).

Acknowledgments

The authors express their gratitude to Dra. Cecilia Carmaran and Dr. José G.

Marmolejo for reviewing the manuscript and useful comments. The authors thank

SEMARNAT - CONACYT (Grant 2002-C01-0409) for their financial support.

R. Valenzuela and T. Raymundo thank COFAA and Instituto Politécnico Nacional

for the financial support for their research in the project SIP-20170845, SIP-

20170846. T. Raymundo expresses her gratitude to CONACYT for the grant of

Postdoctoral stay in the postgraduate of Biosciences of the Instituto Politécnico

Nacional.

New ascomycetes for Sierra de Alamos-Rio Cuchajaqui (Mexico) ... 431

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MY COTAXON

April-June 2017— Volume 132, pp. 433-440

https://doi.org/10.5248/132.433

Dictyotrichocladium aquaticum gen. & sp. nov. and

Minimelanolocus aquatilis sp. nov. from freshwater

in Brazil’s semiarid region

PATRICIA OLIVEIRA Fruza’, Lucas BARBOSA CONCEIGAO’,

Marcos FABIO OLIVEIRA MARQUES’,

Luis FERNANDO PASCHOLATI GUSMAO" & RAFAEL FE, CASTANEDA-RUIZ*

' Universidade Estadual de Feira de Santana, Departamento de Ciéncias Biol6gicas,

Laboratorio de Micologia, Av. Transnordestina, s/n, Novo Horizonte,

44036-900, Feira de Santana, Bahia, Brazil

? Universidade do Estado da Bahia, Departamento de Educacao,

Campus VII, BR 407, Km 127, 48970-000, Senhor do Bonfim, Bahia, Brazil

° Instituto de Investigaciones Fundamentales en Agricultura Tropical (INIFAT) Alejandro de Humboldt,

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, C.P. 17200, Santiago de Las Vegas, C. Habana, Cuba

* CORRESPONDENCE TO: lgusmao@uefs.br

ABSTRACT—A new genus and species, Dictyotrichocladium aquaticum, is described and

illustrated from submerged decaying leaves of Calophyllum brasiliense in the Brazilian semiarid

region; it is distinguished by macronematous, sometimes micronematous conidiophores, and

monoblastic, terminal conidiogenous cells that produce solitary, long pyriform, cylindrical,

long clavate, turbinate to vermiform, curved or sigmoid, dictyoseptate, black or dark

pigmented conidia that usually do not secede but remain attached after maturation. A new

species, Minimelanolocus aquatilis, characterized by obclavate, 3-5-euseptate, verruculose,

pale brown conidia, is described and illustrated from decaying twigs of an unidentified plant

in the Brazilian semiarid region.

KEY worpDs—aquatic fungi, Caatinga, taxonomy, tropics

Introduction

Freshwater hyphomycetes, which are essential for recycling complex organic

matter in aquatic environments (Jones & Pang 2012), have part or entire life

434 ... Fiuza & al.

cycles in water (Thomas 1996). Goh & Hyde (1996) delineated three freshwater

groups: the Ingoldian fungi, the aero-aquatic hyphomycetes, and the facultative

aquatic hyphomycetes. The present study focused on facultative aquatic

hyphomycetes that include hyaline dematiaceous genera producing conidia

that are mainly unmodified by aquatic environments (Goh & Hyde 1996,

Shearer et al. 2007).

In Brazil, the first taxonomic research on facultative aquatic hyphomycetes

occurred in the Brazilian semiarid region (Caatinga) and was followed by

explorations in the Amazon and Atlantic Forest (Barbosa & Gusmao 2011;

Barbosa et al. 2011, 2013; Silva & Gusmao 2013; Monteiro & Gusmao 2014;

Moro et al. 2015; Fiuza et al. 2015a,b, 2016, 2017). These studies revealed a

high biodiversity of facultative aquatic hyphomycetes and a large number of

new taxa.

During studies on freshwater hyphomycetes on submerged decaying plant

material, two interesting taxa were found with conidial morphologies that

differed from those of any described species. One is described here as a new

genus and species, Dictyotrichocladium aquaticum, and the other as a new

species of Minimelanolocus R.F. Castafieda & Heredia, M. aquatilis.

Materials & methods

From November 2013 to August 2015, expeditions were made in Serra da Tromba

(Chapada Diamantina, Bahia) and Serra da Fumaca (Pindobac¢u, Bahia). Both areas

belong to the semiarid region of northeastern Brazil.

In Serra da Tromba, submerged decaying leaves of Calophyllum brasiliense were

collected from three streams in the Contas river basin, taken to the Laboratory of

Mycology (LAMIC) in plastic bags, and processed as described by Barlocher (2005).

Leaf samples of C. brasiliense were washed, and 12 mm diam discs were cut with a cork

borer. The discs were incubated for 48 hours at 18-20°C on a shaker (100 rpm) in 100

mL Erlenmeyer flasks containing 30 mL of sterile distilled water to induce sporulation.

The spore suspensions were filtered through membrane filters (5 um pore size), which

were then mounted on slides with cotton blue in lactic acid. The leaf discs were mounted

on slides with acid lactic and examined under compound microscope.

In Serra da Fumaga, submerged twigs were collected from some streams, taken

to the Laboratory of Mycology (LAMIC) in plastic bags, and processed as described

by Castafieda-Ruiz et al. (2016). The twigs were washed and placed in Petri dishes

(moist chambers) incubated in a polystyrene box with sterile water plus glycerol for

30 days. The slides were mounted using resin PVL (polyvinyl alcohol, lactic acid and

phenol). Measurements of reproductive structures and identification of specimens

were performed using a compound microscope (Olympus BX51). The specimens were

deposited in the Herbarium of Universidade Estadual de Feira de Santana, Bahia, Brazil

(HUEES).

New freshwater fungi from Brazil ... 435

Taxonomy

Dictyotrichocladium Fiuza, Gusmao & R.F. Castafieda, gen. nov.

MycoBank MB 821638

Differs from Trichocladium by its long clavate, turbinate to vermiform, curved or

sigmoid, dictyoseptate conidia.

TYPE SPECIES: Dictyotrichocladium aquaticum Fiuza et al.

ErymMo.oey: Greek, dictyo-, for conidia with one or more transversal and longitudinal

and/or oblique septa; Latin, -trichocladium, referring to the genus Trichocladium.

CONIDIOPHORES micronematous or macronematous, unbranched, septate,

brown. CONIDIOGENOUS CELLS monoblastic, integrated, terminal, pale

brown to brown. CONIDIAL SECESSION schizolytic. Conip1a solitary, long

pyriform, cylindrical, long clavate, turbinate to vermiform, curved or sigmoid,

dictyoseptate, brown to dark brown.

NotEes—Dictyotrichocladium is similar to Trichocladium Harz, which differs by

its didymo- or phragmoconidia (Hughes 1952). Several other genera including

Acrodictys M.B. Ellis, Cirrenalia Meyers & R.T. Moore, Paradictyoarthrinium

Matsush., and Thyrostroma Hohn. have conidia similar to Dictyotrichocladium.

However, Acrodictys produces cylindrical, pyriform, obovoid or ovoid and

straight conidia (Zhao et al. 2011). Cirrenalia is close to Dictyotrichocladium in

conidial shape but differs in its helicoid phragmoconidia (Zhao & Liu 2005). In

Paradictyoarthrinium, the conidiophores are slightly constricted at the septa or

monilioid, and conidiogenous cells are lateral or intercalary (Matsushima 1996,

Seifert et al. 2011). Thyrostroma has stromatic, sporodochial conidiomata, and

its conidiogenous cells are indeterminate with several enteroblastic percurrent

generations (Hohnel 1911). Dictyodesmium S. Hughes and Dictyorostrella

U. Braun also have dictyoseptate conidia, but both genera have stromatic,

sporodochial conidiomata (Seifert et al. 2011).

Dictyotrichocladium aquaticum Fiuza, Gusmao & R.F. Castafieda, sp. nov. Fic. 1

MycoBAnk MB821639

Differs from Trichocladium spp. by its long clavate to vermiform, curved or sigmoid,

dictyoseptate conidia.

Type: Brazil, Bahia: Piaté, Chapada Diamantina, Serra da Tromba, 13°05’S 41°51’W,

alt. 1272 m, on submerged decaying leaves of Calophyllum brasiliense Cambess.

(Calophyllaceae) in a river, 5 June 2015, coll. P.O. Fiuza (Holotype, HUEFS 215954).

EryMo_oey: Latin, aquaticum, referring to the aquatic environment where the species

grows.

COLONIES on natural substrate effuse, brown to dark brown. Mycelium

superficial and immersed, composed of slightly branched, septate, smooth,

436 ... Fiuza & al.

A B

Fic. 1. Dictyotrichocladium aquaticum (holotype, HUEFS 215954). A-F. Conidia; G-I. Conidio-

phores, conidiogenous cells, and conidia. Scale bars = 20 um.

New freshwater fungi from Brazil ... 437

pale brown to brown hyphae, 2-4.5 um diam. CONIDIOPHORES micronematous

or macronematous, single, erect, straight or flexuous, unbranched, 2-8-septate,

smooth, 20-80 x 6-7.5 um, brown. CONIDIOGENOUS CELLS monoblastic,

integrated, terminal, smooth, 6-10 x 7-9 um, pale brown to brown. CONIDIAL

SECESSION schizolytic. Conip1 solitary, long pyriform, cylindrical, long

clavate, turbinate to vermiform, curved or sigmoid, dictyoseptate, smooth,

30-140 x 18-37.5 um, brown to dark brown, dry.

Minimelanolocus aquatilis L.B. Conc., M.F.O. Marques, Gusmao &

R.F. Castaneda, sp. nov. Fic. 2

MycoBank MB821640

Differs from Minimelanolocus limpidus by its smaller conidiophores and verruculose

conidia.

Type: Brazil, Bahia: Pindobagu, Serra da Fumaca, 10°38’S 40°22’W, alt. 865 m, on

submerged decaying twigs of unidentified plant in a stream, 8 August 2015, coll. L.B.

Conceicao (Holotype, HUEFS 216640).

ErymMo_oey: Latin, aquatilis, refers to its capacity or ability to live in water.

COLONIES on natural substrate effuse, hairy, brown. Mycelium superficial and

immersed, composed of slightly branched, septate, smooth, brown hyphae,

2-4 um diam. CONIDIOPHORES macronematous, mononematous, single or

caespitose, erect, straight or flexuous, unbranched, 1—5-septate, think-walled,

smooth, 43-78 x 5-10 um, brown to pale brown at the apex. CONIDIOGENOUS

CELLS polyblastic, integrated, terminal, indeterminate, with several sympodial

extensions, 20-42 x 5-7 um, brown to pale brown. CONIDIAL SECESSION

schizolytic. Conip1A solitary, acropleurogenous, dry, 3-5-euseptate, broadly

obclavate, verruculose, 21-30 x 5-8 um, pale brown to subhyaline, with small

dark scar at the truncate base, 1.5-2 um wide.

Notes—Minimelanolocus currently comprises 29 species (Castafieda-Ruiz et

al. 2001, Heredia et al. 2014, Liu et al. 2015, Hyde et al. 2016), most of which

were found as saprobes on decaying leaves, rotten wood, dead branches, and

bamboo in a terrestrial environment; only five species have been reported from

aquatic habitats (Liu et al. 2015, Hyde et al. 2016). Minimelanolocus aquatilis

is superficially similar to M. limpidus Heredia et al., which has smooth-walled

and longer (27-48 um) conidia (Heredia et al. 2014).

Discussion

Facultative aquatic hyphomycetes have been recorded in Brazil since 2011

(Barbosa et al. 2013). Thus far, 339 species are distributed in Amazon, Caatinga,

and Atlantic Forest (Fiuza et al. 2016, 2017); of these, 25 were described as

438 ... Fiuza & al.

A B cy D A En. F #

fas p i

Fic. 2. Minimelanolocus aquatilis (holotype, HUEFS 216640). A-F. Conidia; G-I. Conidiogenous

cells; J. Conidiogenous cell and conidia; K, L. Conidiophores; M. Conidiophores, conidiogenous

cells, and conidia. Scale bars: A-F, K-M = 10 um; G-J = 5 um.

New freshwater fungi from Brazil ... 439

new species: 15 from Amazon, 1 from Atlantic Forest, and 9 from Caatinga

(Barbosa et al. 2011, 2013; Silva & Gusmao 2013; Monteiro & Gusmao 2014;

Fiuza et al. 2015a,b, 2016; Moro et al. 2015; Conceicao et al. 2016; Monteiro

et al. 2016), including the type species of a new genus, Dictyoaquaphila

J.S. Monteiro et al., from Amazon and Caatinga (Monteiro et al. 2016). The

present study yielded another new species, Minimelanolocus aquatilis and a

new genus, Dictyotrichocladium, to Caatinga.

Acknowledgments

We are grateful to Dr. De-Wei Li and Dr. Jian Ma for critical review of the

manuscript. The authors are grateful to the Programa de Pesquisa em Biodiversidade

(PPBio Semi-arid/MCTI/CNPq—Proc. 457498/2012-9) for financial support.

POF thanks CAPES and LBC is grateful to CNPq for scholarships. POF and LBC

thank Programa de Pés-Graduagao em Botanica (PPGBot/ UEFS). LFPG is grateful

to CNPq for a Proc. 303062/2014-2 grant. RFCR is grateful to Ciéncia Sem Fronteiras

(Proc. 45991/2016-8) and to OSDE, Grupo Agricola from Cuban Ministry of Agriculture

and Programa de Salud Animal y Vegetal (project P131LH003033) for facilities.

Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclatural

review are greatly appreciated.

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MY COTAXON

April-June 2017— Volume 132, pp. 441-444

https://doi.org/10.5248/132.441

Lecanora shangrilaensis sp. nov., on pinecones from China

LEI LU? & ZUN-TIAN ZHAO **

"College of Food Science and Engineering, Qilu University of Technology

Jinan, 250353, China

’ College of Life Sciences, Shandong Normal University

Jinan, 250014, China

* CORRESPONDENCE TO: ztzhao@sohu.com

AsBstTRACT—Lecanora shangrilaensis from southwestern China is described as a new species.

It can be distinguished from other multispored species of Lecanora by the presence of usnic

acid instead of atranorin and the epruinose discs with coarse granular epihymenium. It is

otherwise characterized by the thin thallus, yellow to yellowish brown apothecial disc,

12-16-spored ascus, and the pinecone substrate.

Key worps—East Asia, Lecanoraceae, lichenized Ascomycota, taxonomy

Introduction

Nine multispored species of Lecanora are known: L. bruneri Imshaug

& Brodo, L. cateilea (Ach.) A. Massal., L. japonica Mull. Arg., L. loekoesii

Y. Joshi et al., L. subjaponica L. Li & H.Y. Wang, L. weii L.E Han & S.Y. Guo,

L. pleospora Mill. Arg., L. praesistens Nyl., and L. strobilinoides Giralt &

Gomez-Bolea (Giralt & Gémez-Bolea 1991; Guderley & Lumbsch 1999; Han

et al. 2009; Lui et al. 2011, 2012). A detailed key to these nine species was

presented by Li et al. (2012). Six species have been reported from China:

L. bruneri, L. cateilea, L. japonica, L. loekoesii, L. subjaponica, and L. weii (Han

et al. 2009; Lui et al. 2012; Wang et al. 2007, 2013).

A further study of specimens collected from China has revealed a new

Lecanora species, described here as L. shangrilaensis.

Materials & methods

The specimens studied are housed in the Lichen Section, Botanical Herbarium,

Shandong Normal University, Jinan, China (SDNU) and the Lichen Herbarium,

Liaocheng University, Liaocheng, China (LCU-L).

442 ... Li & Zhao

Thalli were examined and measured under a COIC XTL7045B2 stereo-microscope

and photographed with Olympus SZX16. The anatomical characters (e.g., apothecial

sections, crystalline types, asci, ascospores) were examined by hand-cut sections

under an Olympus CX41 polarizing microscope and photographed with an Olympus

DP72 attached to a Olympus BX61 light microscope. The specimen was chemically

analyzed using spot tests and thin layer chromatography (TLC) with solvent system C

(Orange et al. 2010).

Taxonomy

Lecanora shangrilaensis Z.T. Zhao & L. Lii, sp. nov. PLATE 1

MycoBank MB812187

Differs from other multispored Lecanora spp. by its production of usnic acid instead of

atranorin and its epruinose discs with coarse granular epihymenium.

Type: China. Yunnan Province, Shangrila, Tianshengqiao, alt. 3500 m, on cones of Pinus

sp.. 3 Nov. 2008, Wang 20083559 (Holotype, SDNU; isotype, LCU-L).

ErymMo.ocy: The epithet refers to the county where the species was collected.

THALLUS crustose; surface whitish to yellowish gray, thin, continuous, with

yellowish verruca, esorediate; margin indistinct; prothallus not visible.

APOTHECIA lecanorine, sessile to constricted at the base, 0.2-0.5 mm diam;

disc yellow to yellowish brown, epruinose, plane to slightly concave; margin

concolorous with thallus, thick, prominent, entire, smooth. AMPHITHECIUM

with small crystals soluble in K; cortex distinct, with crystals soluble in

K, basally not thickened, 30-50 um thick; algal layer 20-35 um thick;

PARATHECIUM hyaline; EPIHYMENIUM brown to dark brown, pigment soluble

in K, with coarse crystals soluble in K, 15-20 um thick; HyMENiIum hyaline,

40-50 um thick; HypoTHecium hyaline, 20-45 um thick; SUBHYMENIUM

hyaline, 10-20 um thick; PaRAPHysEs simple, slightly thickened apically,

<2.5 um diam.; Ascr clavate, containing 12-16 spores; AscosporEs hyaline,

simple, ellipsoid, 7.5-12.5 x 5-7.5 um, wall <1 um thick. Pycnip1a not

observed.

CHEMISTRY: cortex K-, C-, KC+ yellow, P-; medulla K-, C-, KC+ yellow,

P-; usnic acid and fumarprotocetraric acid present.

ECOLOGY & DISTRIBUTION: The new species was found on pinecones

from Tianshengqiao, Yunnan Province, southwestern China, at an altitude

of 3500 m.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE, Shangrila,

Tianshengqiao, alt. 3500 m, 3 Nov. 2008, H.Y Wang 20083505, 20083506 (SDNU).

RemarKs: Lecanora shangrilaensis is characterized by a thin thallus,

yellowish discs, coarse granular epihymenium, 12-16-spored ascus, pinecone

Lecanora shangrilaensis sp. nov. (China) ... 443

PiatTE 1. Lecanora shangrilaensis (holotype, SDNU 20083559). a, Thallus. b, Asci. c, Ascospores.

d, Apothecium section. e, Pigment of epihymenium soluble in K. f, Crystals in apothecium section.

g, Crystals of amphithecium and epihymenium soluble in K. Scale bars: a = 500 um; b, c = 10 um;

d-g = 20 um.

substrate, and presence of usnic and fumarprotocetraric acids. The most

diagnostic character for recognizing the new taxon is the presence of usnic

acid instead of atranorin. Lecanora cateilea, L. loekoesii, and L. weii resemble

L. shangrilaensis, but differ in several characters: L. cateilea is distinguished

by heavily pruinose apothecial discs, 8-12-spored asci, and possession of

444 ... Li & Zhao

psoromic acid and atranorin as secondary metabolites; L. loekoesii produces

amphithecia with large crystals and atranorin and lacks phenolic compounds;

and L. weii produces heavily pruinose apothecial discs and a chlarotera-type

epihymenium. Lecanora shangrilaensis can be distinguished from other

multispored species by the presence of usnic acid instead of atranorin and the

yellowish apothecia. Morphologically the new species is similar to L. polytropa

(Ehrh.) Rabenh., which also has a thin thallus and yellowish apothecial discs,

but which has only 8 spores per ascus.

Acknowledgements

The study was financially supported by the National Natural Science Foundation

of China (31300018). The first author thanks SDNU for providing laboratory facilities

to work. The authors also thank Drs. Shouyu Guo (Institute of Microbiology, Chinese

Academy of Sciences, Beijing) and Sergey Kondratyuk (M.H. Kholodny Institute of

Botany, Kiev) for presubmission review.

Literature cited

Giralt M, Gdmez-Bolea A. 1991. Lecanora strobilinoides, a new lichen species from north-eastern

Spain. Lichenologist 23(2): 107-112. https://doi-org/10.1017/S0024282991000270

Guderley R, Lumbsch HT. 1999. Notes on multispored species of Lecanora sensu stricto.

Lichenologist 31: 197-210.

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

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

Lit L, Joshi Y, Elix JA, Lumbsch HT, Wang HY, Koh YJ, Hur JS. 2011. New and noteworthy species

of lichen genus Lecanora (Ascomycota; Lecanoraceae) from South Korea. Lichenologist 43(4):

321-329. https://doi.org/10.1017/S0024282911000144

LiL, Zhang LL, Liu XL, Zhao ZT, Wang HY. 2012. Lecanora subjaponica, a new lichen from China.

Lichenologist 44(4): 465-468. https://doi.org/10.1017/S002428291200014X

Orange A, James PW, White FJ. 2010.Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

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

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

Wang HY, Ge AN, Li HM, Zhao ZT. 2013. Additional information on Lecanora loekoesii. Mycotaxon

123: 235-239. https://doi.org/10.5248/123.235

MY COTAXON

April-June 2017— Volume 132, pp. 445-451

https://doi.org/10.5248/132.445

Cephaliophora tropica: a third European record

MALGORZATA RUSZKIEWICZ-MICHALSKA’ , PIOTR KNYSAK’,

IZABELA SKROBEK’, ANETA GWIAZDA?’, SEBASTIAN PISKORSKI* &

JOANNA ZELAZNA-WIECZOREK'’

' Department of Algology and Mycology & * Myco-Algological Section of the Students Society,

Faculty of Biology and Environmental Protection, University of Lodz,

Banacha 12/16, PL-90-237 LédZ, Poland

" CORRESPONDENCE TO: mrusz@biol.uni.lodz.pl

ABSTRACT—We describe the morphology of Cephaliophora tropica based on a specimen

retrieved from samples of urban water bodies in Ldédz city, central Poland, and isolated by

a shed snakeskin bait method. We also review the species’ distribution to demonstrate its

circumglobal range. With previous records from Hungary and Finland, this is only the third

report of C. tropica from Europe.

KEY WORDS—aquatic environment, conidial fungi, distribution, hyphomycetes, Pezizales

Introduction

Cephaliophora Thaxt. (Pezizales, Ascomycota) encompasses hyphomycete

species without known teleomorphs (Kirk et al. 2008). Thaxter (1903) originally

erected the genus for two coprophilous species: C. tropica (type species) and

C. irregularis Thaxt. [= C. nigricans (Bainier) Bainier]. A third species accepted

in the genus is C. uniformis Narayanan, isolated from jute sacks (Narayanan

1962). Three rotifer- and tardigrade-capturing species were also described in

Cephaliophora as C. longispora G.L. Barron et al., C. muscicola G.L. Barron et

al., and C. navicularis Tzean & G.L. Barron (Tzean & Barron 1983, Barron et

al, 1990), but it has been suggested that they belong in Lecophagus (Dick 1990,

Tanabe et al. 1999).

Seifert et al. (2011) refer to C. tropica as a pantropical and occasionally

temperate species. Our survey of literature citations of C. tropica indicates

446 ... Ruszkiewicz-Michalska & al.

TABLE 1. Literature review of Cephaliophora tropica reports

COUNTRY HOST/SUBSTRATE * REFERENCE

Africa

Central African Republic Dung (Bovidae) CBS 139.59

Egypt Air-borne dust Abdel-Hafez et al. (1990)

Khallil & Abdel-Sater (1992)

Moth pupae (Noctuidae) Ismail & Abdel-Sater (1993)

Ghana Theobroma (Malvaceae) Farr & Rossman (2016)

Liberia Dung (Equidae) Thaxter (1903)

Asia

China Dung (Muridae) Thaxter (1903)

Endophytic (Salicaceae) Zhao & Li (2012)

India Dung (Cercopithecidae) Iyer et al. (1973)

Soil Barron (1968)

Indonesia Dung (Muridae) Thaxter (1903)

Iran Soil Pordel et al. (2015)

Israel Trigonella (Fabaceae) Chabelska-Frydman (1959)

Japan Vermicompost Huang et al. 2013

; : Moustafa (1986

PBA ei ie ie 1990)

Pakistan Helianthus (Asteraceae) Shahzad & Ghaffar (1995)

Sharfun-Nahar & Hashmi (2005)

Mukhtar (2010)

Mushtaq & Hashmi (2005)

Ricinus (Euphorbiaceae) Dawar et al. (2014)

Diverse substrates Ellis (1971)

Russia Rhizosphere (Sapindaceae) Zlotnikov et al. (2007)

Thailand Dung (rodents) Suwanarit (1986)

Turkey Capsicum (Solanaceae) CBS 996.69

Australia & Oceania

Australia Diverse substrates Ellis (1971)

Painted steel panels; Crook & Hindson (1995)

organic materials

Solomon Is. Diverse substrates Ellis (1971)

Europe

Finland Wooden parts (sauna) Salonen & Ruokola (1969)

Hungary Dung (Cervidae) Toth (1994)

Révay (1998)

North & Central America

Barbados Soil Barron (1968)

Jamaica Dung (Herpestidae) cee 001)

Mexico Vachellia (Fabaceae) Ale-Agha et al. (2007)

U.S.A. Roystonea (Arecaceae) Farr & Rossman (2016)

Spathiphyllum (Araceae)

Helianthus (Asteraceae) Roberts et al. (1986)

Nicotiana (Solanaceae) Wolf (1949)

South America

Venezuela Dung (animals; incl. parrots) Delgado Avila et al. (2002)

* Plant classification acc. to Stevens (2012)

Cephaliophora tropica in Poland ... 447

its circumglobal distribution (TaBLe 1), showing the most numerous data

originating from Asia with no reports from Arctic and Antarctic. The only

European records are from Finland (Salonen & Ruokola 1969) and Hungary

(Révay 1998, and literature cited therein).

Materials & methods

In 2015, a study of fungal parasites of planktonic algae was conducted in urban,

eutrophic aquatic reservoirs, including the artificial flow pond Mlynek on the Olechéwka

river in Lodz, Central Poland (51°43°32"N 19°31’28"E). The river, which flows mainly

within the city limits, also supplies several other artificial flow ponds. The Mlynek

pond (370 x 30-95 m) is part of an urban park system and in close proximity to a large

residential area. The pond is used mainly for recreation, but no swimming is allowed.

On 21 July 2015 a water sample was collected from the shoreline of the pond and

held in 125 ml plastic containers. Physical parameters measured included conductivity

(EC = 483.1 uS/cm), pH (= 8.2), air temperature (= 25.8°C), and water temperature

(=25.7°G):

The sample was transported in an insulated container to the laboratory of the

Department of Algology and Mycology, University of Lodz. A bait method was used in

order to isolate fungi in the lab (Batko 1975). Three baits (sesame seeds, shed snakeskin,

onion peels) were sterilized with hypochlorite solution and UV irradiation in a laminar-

flow cabinet; they were distributed onto sterile Petri dishes together with 20 ml of water

sample and incubated for 30 days at 24 + 2°C. Fungi were identified according to Ellis

(1971) with the aid of a Nikon SMZ 745T stereomicroscope and Nikon E-400 light

microscope (semi-solid preparations) and photographed using a Nikon DS-F1 camera.

Taxonomy

Cephaliophora tropica Thaxt., Bot. Gaz. 35: 158 (1903) PLATE 1

COLONIES appearing light pink to reddish brown from the colour of conidial

mass, effuse. Hyphae superficial, colourless, thin-walled, septate, 5.0-7.5 um

diam. CONIDIOPHORES macronematous, with terminal or rarely lateral ampulla,

clavate, straight or slightly flexuous, colourless, smooth, 2-3-celled, 24-30 x

18-20 um; basal cell trapezoidal, in 3-celled conidiophores the middle cell often

slightly sloping. CONIDIOGENOUS CELLS terminal, subsphaerical to clavate,

19-29 x 16-20 um, bearing up to 3 mature and tens of smaller immature

conidia visible as balloon-like outgrowths at the surface of the conidiogenous

cell. Conip1a forming simultaneously, solitary, mostly 2-3-septate, 18-24 x

24-37 um; cylindrical or slightly clavate, basal cell pointed, arising from a

prominent wide (<2.5 um diam.) hilum; pale to cinnamon brown, basal and

topical cells paler than the central ones, in some mature spores basal cell almost

colourless; septa darker, brown, <2 um thick.

448 ... Ruszkiewicz-Michalska & al.

PLATE 1. Cephaliophora tropica (LOD AF-33). A: Mature and immature conidiophores and conidia.

B1-3: Conidiophores and conidiogenous cells; black arrow (B1) pointing to young two-celled

conidium; white arrow (B2, B3) pointing to the youngest conidia visible as balloon-like outgrowths

on the conidiogenous cell. C: mature conidia. Scale bars = 10 um.

SPECIMEN EXAMINED—POLAND: L6pz ciry, Mlynek artificial flow pond, shore area,

21 July 2015, leg. P. Knysak (LOD AF-33).

Discussion

The size of the conidiophores and conidia reported by us match the lower

limits of measurements given in other reports of the species. In conidia

formed in the water-derived culture, the maximum number of septa was

four. Similarly, Thaxter (1903) and Wolf (1949) reported 2-5-septate (mainly

3-septate) conidia, although subsequent cultural studies by Mayoral et al.

(1964) and Weik & Pappelis (1964) noted conidia with up to 8 and 9 septa.

Cephaliophora tropica in Poland ... 449

Although both of the latter studies reported conidia that were deformed or

with atypical shapes, we have observed only regular conidial forms.

Beside the two main substrates of C. tropica (e.g., dung and soil), the

diversity of reported substrata covers a range of living and dead plant and

animal tissues (TABLE 1) as well as air-borne dust and a painted steel surface.

To our knowledge, we are the first to isolate the species from a sample derived

from an aquatic ecosystem. Although Kaushal & Singh (2012) reported

C. tropica from areas surrounding water bodies in India, their photograph of

the fungus does not contain necessary information to confirm the identification

as C. tropica.

The ability to utilize diverse carbon and nitrogen sources was studied by

Mayoral et al. (1964) and Weik & Pappelis (1964). Cephaliophora tropica

grew well using all nine tested nitrogen-containing compounds, but fungal

sporulation was enhanced by nitrate-N (Weik & Pappelis 1964). Cellobiose

and hexoses (fructose, glucose, galactose, and mannose) proved to be the best

carbon sources as indicated by mycelium dry weight and change of pH of

cultural media (Mayoral et al. 1964). In our experiment, colonies of C. tropica

developed only on shed snakeskin, suggesting that that substrate had a positive

influence on growth. However, species preference toward keratin is still to be

tested.

Acknowledgments

The authors are deeply grateful to reviewers Prof. Maria Dynowska (University of

Warmia and Mazury, Olsztyn, Poland), Prof. Harish C. Gugnani (St. James School of

Medicine, Dutch Caribbean), and MSc. Patricia Oliveira Fiuza (Universidade Estadual

de Feira de Santana, Brazil) for critical and constructive comments on the manuscript.

The study was financially supported in frame of the statutory funds of Department of

Algology and Mycology, University of Lodz, in the years 2015-16.

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MY COTAXON

April-June 2017— Volume 132, pp. 453-458

https://doi.org/10.5248/132.453

Tretoheliocephala compacta gen. & sp. nov.

from the Brazilian semi-arid region

Luis FERNANDO PASCHOLATI GUSMAO’,, JOSIANE SANTANA MONTEIRO’,

& RAFAEL F. CASTANEDA-RUIZ?

' Universidade Estadual de Feira de Santana,

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

? Instituto de Investigaciones Fundamentales en Agricultura Tropical (INIFAT) Alejandro de Humboldt,

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, C.P. 17200, Santiago de Las Vegas, C. Habana, Cuba

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—A new genus and species, Tretoheliocephala compacta, found on decaying leaves

of an unidentified plant, is described and illustrated. It is characterized by macronematous,

erect dark brown conidiophores with a compact cluster of globose, monotretic conidiogenous

cells, which produce solitary or blastocatenate, ovoid, oblong, dolabriform, sub-obpyriform

or obcuneiform, 0—2-septate, brown conidia.

Key worps—Ascomycota, hyphomycetes, taxonomy, tropics

Introduction

The diversity of conidial fungi in the Brazilian semi-arid region has received

little attention. However, recent studies conducted in Caatinga biome reveal a

high number of new microfungal species (Almeida et al. 2016, Conceigao et al.

2016, Monteiro et al. 2016, Silva et al. 2016). During a survey of hyphomycetes

associated with plant materials from Bahia state, a conspicuous fungus was

collected that showed remarkable differences from all previously described

genera (Seifert et al. 2011) and is described here as new.

Materials & methods

Samples of leaf litter were placed in plastic bags for transport to the laboratory,

where they were placed in Petri dish moist chambers and stored at 25°C for 30 days in

454 ... Gusm4o, Monteiro & Castafieda-Ruiz

a 170 L polystyrene box with 200 mL sterile water plus 2 mL glycerol (Castafieda-Ruiz

et al. 2016). Mounts were prepared in PVL (polyvinyl alcohol and lactic acid) and

measurements were made at x1000 magnification. Micrographs were obtained with

an Olympus BX51 microscope equipped with bright field and Nomarski interference

optics. The specimens, including the holotype, are deposited in the Herbarium of

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

Taxonomy

Tretoheliocephala Gusmao, J.S. Monteiro & R.F. Castafieda, gen. nov.

INDEXFUNGORUM IF553563

Differs from Heliocephala by its tretic conidial ontogeny.

TYPE SPECIES: Tretoheliocephala compacta Gusmao et al.

EryMo.Locy: Latin, treto-, referring to the tretic conidial ontogeny +

-heliocephala, referring to the genus Heliocephala.

Conidial fungi. CoLonies on the natural substratum effuse, hairy, dark

brown or black. CONIDIOPHORES macronematous, mononematous, erect,

unbranched or branched, cylindrical below, capitate at the apex, septate, with a

distinct smooth or verruculose, brown or dark brown. CONIDIOGENOUS CELLS

monotretic, discrete, globose, determinate, brown to dark brown, forming

a more or less spherical cluster. Conidial secession schizolytic. CONIDIA

solitary or blastocatenate, unicellular or euseptate, ovoid, oblong, dolabriform,

subobpyriform or obcuneiform, smooth, pale brown or brown.

Tretoheliocephala compacta Gusmao, J.S. Monteiro & R.F. Castafieda, sp. nov.

PIGS:192

INDEXFUNGORUM IF553564

Differs from Heliocephala spp. by its monotretic conidiogenous cells.

Type: Brazil, Bahia: Mucugé, Parque Nacional da Chapada Diamantina, 13°00’S

41°22’W, on decaying of unidentified plant, 10 January 2005, coll. L.EP. Gusmao

(Holotype, HUEFS 81045).

EryMo_oey: Latin, compacta, referring to the compact arrangement of conidiogenous

cells in the capitulum.

Co.ontgs on the natural substratum effuse, hairy, amphigenous, dark brown

or black. Mycelium mostly immersed, composed of septate, smooth, brown

hyphae, 1-2.5 um diam. CONIDIOPHORES macronematous, mononematous,

cylindrical below, capitate at the apex, erect, straight, robust, with 1-3

enteroblastic percurrent extension, 7-10-septate, unbranched or branched

above, smooth below, somewhat verrucose or with remains of the old

collapsed conidiogenous cells embedded near of each enteroblastic percurrent

Tretoheliocephala compacta gen. & sp. nov. (Brazil) ... 455

egabahbse

Fic. 1. Tretoheliocephala compacta (holotype, HUEFS 81045). A. Conidia. B. Clusters of

conidiogenous cells and conidia. C. Conidiophores, conidiogenous cells, and conidia.

D. Conidiogenous cells. E, EF Conidiogenous cells and conidia. Scale bars: A, D-F = 10 um;

B = 20 um; C = 50 um.

456 ... Gusmao, Monteiro & Castafieda-Ruiz

Fic. 2. Tretoheliocephala compacta (holotype, HUEFS 81045).

Conidiogenous cells and conidia. Scale bar = 10 um.

regeneration of the conidiophores, black below, dark brown or brown near the

conidiogenous capitulum, 320-700 x 15-19 um; branches black, 200-300 x

13-15 um. CONIDIOGENOUS CELLS monotretic, discrete, globose, dark brown

or brown, smooth, determinate, 8-12 um diam, disposed in a compact cluster

forming of the more or less spherical capitulum. Conidial secession schizolytic.

ConipIia solitary or blastocatenate, ovoid, oblong, dolabriform, subobpyriform

or obcuneiform, 0-2-septate, mostly 1-septate, 12-15(-21) x 6-7 um, pale

brown, brown or dark brown, sometimes with the basal cell paler than the

apical cell, black at septa.

Tretoheliocephala compacta gen. & sp. nov. (Brazil) ... 457

ComMENtTs: Heliocephala V. Rao et al. resembles Tretoheliocephala, but

Heliocephala is characterized by conidiophores that are macronematous,

mononematous, brown, solitary, erect, straight or flexuous, with short branches

and bearing terminal clusters of conidiogenous cells or elongated percurrently

and bearing discrete clusters of conidiogenous cells along their lengths.

Heliocephala also produces conidiogenous cells that are monoblastic, hyaline

to light brown, ampulliform to lageniform borne on the short branches and

conidia that are obclavate to navicular, subhyaline to olivaceous, 1-3-septate,

rostrate, sometimes germinating at the rostrum to produce secondary clusters

of conidiogenous cells (Decock et al. 1998, Heredia et al. 2011, Kumaresan &

Srinivasan 2002, Melnik et al. 2013, Rao et al. 1984). These two genera are

easily separated.

Acknowledgments

We are indebted to Dr. Jian Ma and Dr. De-Wei Li for critical review. The

authors thank the National Council for Scientific and Technological Development

(CNPq) (Proc. 451991/2016-8). The first author is grateful to Coordination for

the Improvement of Higher Education Personnel (CAPES) through project N°

071/2012. RFCR is grateful to the OSDE, Grupo Agricola from the Cuban Ministry

of Agriculture and Programa de Salud Animal y Vegetal (project P131LH003033). Dr.

Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature review

are greatly appreciated.

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Mycotaxon 123: 281-284. https://doi.org/10.5248/123.281

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Dictyoaquaphila appendiculata gen & sp. nov. from submerged wood from Brazil. Mycotaxon

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Diamantina, Bahia, Brazil. Mycotaxon 130: 1125-1133. https://doi.org/10.5248/130.1125

MY COTAXON

April-June 2017— Volume 132, pp. 459-470

https://doi.org/10.5248/132.459

Sporormiella longicolla sp. nov. and

new Sporormiella records on herbivore dung from Brazil

ROGER FAGNER RIBEIRO MELO **, ANDREW N. MILLER ”

& LEONOR CosTA MAIA '

"Universidade Federal de Pernambuco, Departamento de Micologia,

Av. da Engenharia, s/n, 50740-600, Recife, Brazil

? University of Illinois at Urbana-Champaign, Illinois Natural History Survey,

1816 S. Oak St., Champaign, IL 61820, USA

“ CORRESPONDENCE TO: rogerfrmelo@gmail.com

ABSTRACT—Seven species of Sporormiella are reported from fresh dung collected in a

survey including different vegetation areas throughout Pernambuco, Northeastern Brazil.

An undescribed, long-necked, small-spored species is proposed as Sporormiella longicolla.

Sporormiella isomera is a new record for the Neotropics; S. australis, S. leporina, and

S. pentamera are new records for Brazil; and S. herculea (a rare species) and S. minima are

also recorded. Descriptions, photographic plates and digital line drawings are provided,

along with an identification key to all Sporormiella species recorded in Brazil. Substrate

relationships and distribution data of these fungi are also discussed.

Key worps—Ascomycota, coprophilous fungi, Pleosporales, Sporormiaceae, taxonomy

Introduction

Sporormiella Ellis & Everh. 1892 comprises ~80 pseudothecial species,

usually with dark, 4-celled ascospores, commonly found on herbivore

dung worldwide (Kirk et al. 2008). The genus is characterized by small,

dark brown, glabrous or hairy uniloculate pseudothecia, bitunicate asci,

cylindrical to clavate, bearing eight three-to many-septate, dark brown

ascospores with conspicuous germ slits, surrounded by a hyaline gelatinous

sheath (Ahmed & Cain 1972). Species of Sporormiella are predominantly

coprophilous (Richardson 2001b), with occasional records as saprobes on

460 ... Melo, Miller & Maia

other substrata, and even as endophytes (Sun et al. 2006). Recently, the

genus has received attention as a paleontological indicator to trace the

wave of human invasion, habitat destruction, and subsequent megafaunal

extinctions in Quaternary lake deposits (Comandini & Rinaldi 2004, Raper

& Bush 2009).

The monophyly of Sporormiella, as well as other members of

Sporormiaceae Munk, was studied by Kruys & Wedin (2009), who suggested

a new generic classification of the family, in which Preussia Fuckel would

include Sporormiella and Spororminula Arx & Aa. Although Kruys &

Wedin (2009) argued for the synonymy of Preussia and Sporormiella, they

demonstrated no significant support for a joint monophyletic grouping

and stated that resolution of the Preussia—Sporormiella complex required

further study.

In Brazil, there have been few concentrated surveys on coprophilous

mycobiota, and little is known about Sporormiella. After a visit to Brazil

in 1998, Richardson (2001la) recorded S. minima and S. cf. megalospora

(Auersw.) S.I. Ahmed & Cain in Bonito and Pantanal do Rio Negro,

Mato Grosso do Sul State, providing short descriptions and discussing

the importance of further investigations. Here, the Sporormiella species

recorded on dung from Brazil are revisited, including revision of herbaria

material, and new records and an undescribed species from fresh material

collected in Pernambuco State.

Material & methods

Dung samples of camel (Camelus bactrianus), deer (Cervus elaphus), llama (Lama

glama), and waterbuck (Kobus ellipsiprymnus) were collected from August 2009 to

July 2010 in a zoological park in Recife (8°07’30"S 34°52’30”W), and samples of cattle

(Bos taurus), goat (Capra hircus) and horse (Equus caballus) dung were collected

from animal precincts on the campus of Universidade Federal Rural de Pernambuco,

Recife (8°00'54”S 34°56’59”W), and from farms close to the Instituto Agronémico de

Pernambuco (IPA) in Caruaru (8°01’59”S 36°06’59” W) and Serra Talhada (7°54’59”S

38°17'00’W), all located in Pernambuco, Northeastern Brazil. The samples were

collected with sterilized spatulas, placed in clean plastic bags, taken to the laboratory

in less than 24 hours and incubated in moist chambers at room temperature

(28 + 2 °C) for at least 60 days under alternating natural light and dark periods.

The specimens were observed directly from the substrata under a Leica EZ4

stereomicroscope. Pseudothecia were mounted in tap water and/or lactophenol with

cotton blue for measurements and identification, and in Polyvinyl-Lacto-Glycerol

resin for preservation. Specimens were identified based on morphology according

to Ahmed & Cain (1972), Bell (1993, 2005), Richardson & Watling (1997), and

Sporormiella longicolla sp. nov. (Brazil) ... 461

Doveri (2004). A survey of representative national collections of fungi (acronyms

following Thiers 2016) was also conducted, including the herbaria Pe. Camille Torrend

(URM), Dardano de Andrade Lima (IPA), Maria Eneyda P. Kaufmann Fidalgo (SP),

Dimitri Sucre Benjamin (RB), Instituto Nacional de Pesquisas da Amazonia (INPA),

Museu Paraense Emilio Goeldi (MG), and Instituto de Biociéncias, Universidade

Federal do Rio Grande do Sul (ICN). High quality images were captured with a

QImaging QColor 3 digital camera mounted on an Olympus BX51 compound

microscope using differential interference or phase contrast microscopy. Methods for

the digital line drawing illustrations of both pseudothecia and microscopic structures

were adapted from Barber & Keane (2007). Permanent slides were mounted

and deposited in Herbario Padre Camille Torrend, Departamento de Micologia,

Universidade Federal de Pernambuco, Recife, Brazil (URM).

Taxonomy

Sporormiella australis (Speg.) S.1. Anmed & Cain,

Canad. J. Bot. 50: 434 (1972) Fic. 1A—pD

PSEUDOTHECIA scattered, immersed when young, becoming partially

immersed to superficial on dung when mature, subglobose to obpyriform,

dark brown to black, 185-210 x 145-175 um diam. NECK small, papilliform,

glabrous, black. PERIDIUM membranaceous, semitransparent, composed of

angular cells. Asc bitunicate, 8-spored, cylindrical to clavate, broader near

the apex, rounded above, tapering abruptly downwards to form a short stipe,

125-140 x 14.5-18 um. Ascospores obliquely triseriate, less commonly

biseriate, 4-celled, cylindrical, rounded at the ends, light brown to brown,

37.5-40 x 6-7.5 um, smooth, with easily separable segments in all septa, germ

slits oblique, surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO, Caruaru, Instituto Agrondémico

de Pernambuco (IPA), on cattle dung, 6.VI.2012, RLER. Melo (URM86773); Recife,

Horto Zoobotanico do Parque Dois Irmaos, on goat dung, 12.1X.2011, R.ER. Melo

(URM86775).

Notes: Sporormiella australis can be easily mistaken for other common

Sporormiella species based on ascospore shape, which are smaller in

S. minima (27.5-30 x 4-5 um) and larger in S. intermedia (Auersw.)

S.I. Ahmed & Cain (48-59 x 9.5-11.5 um) (Ahmed & Cain 1972). The

ascospore cells of S. australis do not easily separate in all septa.

Sporormiella herculea (Ellis & Everh.) $.1. Anmed & Cain,

Canad. J. Bot. 50: 442 (1972) Fic. 1E-1

PSEUDOTHECIA scattered, immersed on dung, obpyriform, dark brown

to black, 210-275 x 130-155 um diam. NEcK cylindrical, glabrous, black.

462 ... Melo, Miller & Maia

PERIDIUM membranaceous, semitransparent to opaque, composed of

angular cells. Ascr bitunicate, 8-spored, cylindrical-clavate, with somewhat

narrowed apex, tapering abruptly below to form a short stipe, 235-270 x

45-50 um. Ascospores obliquely uni- to biseriate, 6-12-celled, cylindrical

to fusiform, with a wide morphological diversity, rounded or narrowed at

the ends, with some cells larger and thicker than the others, usually with the

uppermost spore in the ascus bearing a greatly enlarged and pigmented cell,

dark brown at maturity, 87.5-112 x 15-20(-25) um, smooth, with transverse

germ slits, surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto Zoobotanico do

Parque Dois Irmaos, on horse dung, 9.1.2012, R.LER. Melo s.n. (URM86777); Serra

Talhada, Instituto Agronédmico de Pernambuco, on horse dung, R.ER Melo s.n.

(URM86776).

Notes: Sporormiella herculea is distinguished by its 6-16-celled ascospores,

with the second to fifth cell sometimes greatly enlarged in the uppermost

spore in the ascus. Ahmed & Cain (1972) discussed morphological differences

of Brazilian records.

Sporormiella isomera S.I. Ahmed & Cain, Canad. J. Bot. 50: 445 (1972) Fig. 1J-L

PSEUDOTHECIA scattered, immersed when young, becoming partially

immersed on dung when mature, subglobose, dark brown to black, 190-

245 um in diam. NeEcK small, papilliform, glabrous, black. PERIDIUM

membranaceous, semitransparent, composed of angular cells. Asc 8-spored,

cylindrical-clavate, broader near the apex, tapering gradually downwards to a

short stalk, 130-150 x 12.5-15 um. Ascosporegs obliquely biseriate, 4-celled,

cylindrical, rounded at the ends, light brown when young, becoming dark

brown at maturity, 35-40 x 5-7 um, smooth, with easily separable segments

in all septa, with germ slits parallel to oblique, with a crook near the middle,

surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto Zoobotanico do

Parque Dois Irmaos, on camel dung, 5.IV.2010, R.ER. Melo s.n. (URM82397,

82398).

Notes: Sporormiella isomera is similar to S. leporina, except it possesses

easily separable rounded terminal ascospore cells with a germ slit crooked

near the middle. Sporormiella minima differs in having smaller ascospores

(27.5-30 x 4-5 um) and pseudothecia (115-165 x 90-125 um) and asci that

are abruptly constricted to form a stipe at their base.

Sporormiella longicolla sp. nov. (Brazil) ... 463

Sporormiella leporina (Niessl) S.1. Ahmed & Cain,

Canad. J. Bot. 50: 447 (1972) Fic. 1M,N

PsEUDOTHECIA isolated to scattered, immersed when young, becoming

partially immersed on dung when mature, subglobose, dark brown to

black, 180-195 x 150-170 um. Necx cylindrical, glabrous, black. PERIDIUM

membranaceous, semitransparent, composed of angular cells. Asc 8-spored,

cylindrical-clavate, with a short stalk, 95-125 x 12-17.5 um. ASCOSPORES

obliquely biseriate, 4-celled, cylindrical, narrowed at the ends, light brown

when young, becoming dark brown at maturity, 30-36.5 x 5-6.5 um,

smooth, equally separable in all segments, with germ slits strongly oblique,

surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto Zoobotanico do

Parque Dois Irmaos, on llama dung, 02.11.2010, R.ER. Melo s.n. (URM82395, 82396).

Notes: Sporormiella leporina can be distinguished by its cylindrical-clavate

asci that are wider near the apex and gradually narrow toward the base and

its ascospore cells with oblique germ slits.

Sporormiella longicolla R.ER. Melo, sp. nov. Fics 2, 3

MycoBank MB815023

Differs from 4-celled Sporormiella species by its longer neck, longer and narrower asci,

and ascospores with persistently united cells.

Type: Brazil. Pernambuco: Recife, Horto Zoobotanico do Parque Dois Irmaos, on horse

dung, 29.11.2012, R.ER. Melo s.n. (Holotype, URM87584).

EryMo oey: longicolla = with a long neck.

PsEUDOTHECIA isolated to gregarious, immersed to semi-immersed on the

substrate, subglobose to obpyriform, dark brown to black, 450-650 x 140-220

um, glabrous. NECK cylindrical, long, glabrous, carbonaceous, black, 250-460(-

550) x 40-65 um, straight or flexuous. PERIDIUM pseudoparenchymatous,

membranaceous, composed by angulated, thick-walled cells, 6-12 um long.

Asct bitunicate, 8-spored, cylindrical-clavate, tapering slightly downwards to

form a long stipe, twice the length of the sporiferous part, (72-)95-105(-137.5)

x 7.5-10 um. Ascospores obliquely biseriate, 4-celled, slightly apiculate on

both ends, hyaline when young, becoming brown to dark brown when mature,

17.5-20.5 x 3-3.5 um, smooth, usually with the cells remaining united even

after the spore liberation, with longitudinal germ slit, surrounded by a thin

hyaline gelatinous sheath. Anamorph unknown.

ADDITIONAL MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto

Zoobotanico do Parque Dois Irmaos, on horse dung, 15.11.2012, R.RR. Melo s.n.

(URM87585).

464 ... Melo, Miller & Maia

Notes: Sporormiella longicolla has pseudothecia with long prominent necks,

long slender asci with stipes c. 2x the length of the sporiferous part, and

small stout ascospores with cells that remain united after spore liberation.

This combination of morphological characters supports separation of a new

species.

Sporormiella minima (Auersw.) S.I. Ahmed & Cain,

Pakistan J. Sci. Industr. Res. 12: 241 (1970) Fic. 1o-s

PsEUDOTHECIA isolated to gregarious, immersed to semi-immersed,

subglobose to obpyriform, dark brown, 115-165 x 90-125 um. NECK

papilliform, glabrous, black, 45-50 x (35-)40-50 um. Ascr bitunicate,

8-spored, cylindrical, with the lower portion abruptly constricted to form

a short stipe, 90-97.5 x 12.5-15 um. Ascospores obliquely biseriate to

triseriate, 4-celled, rounded at the ends, hyaline when young, becoming

brown to dark brown, 27.5-30 x 4-5 um, smooth, easily separable at the

central septum, with germ slits parallel to the spore axis, with a crook at the

central portion on each cell, surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto Zoobotanico do

Parque Dois Irmaos, on llama dung, 13.X.2009, R.ER. Melo (URM82389); 2.1I.2010,

R.ER. Melo (URM82390); 11.2010, R.ER. Melo (URM82391); 25.III.2010

(URM82392); 5.IV.2010 (URM82393); 2.V1.2010 (URM82394); on guinea pig dung,

1.X11.1947 (IPA37901).

Notes: Sporormiella minima is the most common Sporormiella species on

herbivore dung from Brazil. Its striking features include the short ascus stipe

formed by an abrupt end at its base, small ascospores (27.5-30 x 4-5 um

in the examined material) with a tendency to separate only at the central

septum (even inside the asci), and parallel germ slits that form a distinct

crook at the middle. Exsiccata IPA 37901 contained, along with the holotypus

of Ascobolus cuniculorum Bat. & Pontual, several pseudothecia of S. minima,

well preserved on dry guinea pig dung.

Sporormiella pentamera (Oudem.) S.I. Ahmed & Cain,

Canad. J. Bot. 50(3): 454 (1972) Fig 1T-v

PsEUDOTHECIA Scattered to isolated, immersed to partially immersed on

dung, subglobose, dark brown to black, 190-215.5 x 30-50 um. NECK small,

papilliform, glabrous, black. Asci 8-spored, clavate, with rounded apex,

broader near the middle, tapering slightly downwards to form a short stipe,

190-210 x 15-17 um. Ascosporss biseriate, 5-celled, cylindrical, rounded

to slightly narrowed at the ends, light brown when young, becoming dark

Sporormiella longicolla sp. nov. (Brazil) ... 465

Fic. 1. Sporormiella species recorded on herbivore dung from Brazil. S. australis (U

RM86773):

A. pseudothecium on dung; B. pseudothecium in mountant; C. mature ascus with cells united or

equally separated on each septum; D. ascospore cell with a visible germ slit. S$. herculea (URM86777):

E. pseudothecium on dung; FE, G. mature asci; H, I. ascospores. S. isomera (U

RM82398):

J. pseudothecium in mountant; K. ascospores equally separated in all septa; L. mature ascus.

S. leporina (URM82395): M. pseudothecium in mountant; N. mature ascus. S. minima (U

O. pseudothecium on dung; P. pseudothecium in mountant; Q. mature ascus; R, S.

RM82391):

ascospores,

usually separated in the middle septum. S. pentamera (URM86786): T. pseudothecium on dung;

U. pseudothecium in mountant; V. ascospores. Scale bars: A, B, F, G, U = 50 um; C = 25

um; D, R, S

= 5 um; E = 300 um; H, I, L, M, P = 40 um; J, O = 100 um; K, Q= 10 um; N, V = 15 um; T = 150 um.

466 ... Melo, Miller & Maia

Fic. 2. Sporormiella longicolla (holotype, URM87584): A. pseudothecium on dung;

B. pseudothecium in mountant; C. mature ascus; D. young ascus; E. ascospores with united cells

even after their liberation; F. ascospore cells with visible germ slits (arrow). Scale bars: A = 300

um; B = 60 um; C = 7.5 um; D = 15 um; E = 10 um; F = 2.5 um.

Sporormiella longicolla sp. nov. (Brazil) ... 467

Fic. 3. Sporormiella longicolla (holotype, URM87584): A. pseudothecium on dung. B. long-

necked pseudothecia. C. young ascus . D. mature ascus. E. ascospores with united cells even

after their liberation. F. ascospore cell with a visible germ slit. Scale bars: A = 1 mm; B = 50 um;

C= 10 um; D, E=5 um; F= 2 um.

468 ... Melo, Miller & Maia

brown at maturity, 65-72.5 x 18-20.5 um, usually with the second cell from

the upper end of the spore larger than the others, smooth, with segments not

easily separable, germ slits oblique, surrounded by a gelatinous sheath.

MATERIAL EXAMINED: BRAZIL. PERNAMBUCO: Recife, Horto Zoobotanico do

Parque Dois Irm4os, on horse dung, 13.X1.2011, R-LER. Melo (URM86786).

Key to species of Sporormiella on herbivore dung from Brazil

WAscosporesAacelled. <\.n24 <2 9% 2 ee 4 ehag 08 eho eg e Yale oF ela ey age oe age eM ge 2

PVOOOSPOLESeerrcel le 5.x. teel fies cten! ize: apa nae coreg (Peewee apace, «Baaoe oh soe Adee Ba 3

2. Neck >250 um long; ascospores 17.5-20.5 um long .................. S. longicolla

Necks <2 00: (ny ASCOSPOTes SIS MIT TOTS AF oat tea Tyar steer Pyponighath sight corsa sori hdl Sle 7

3. Asci tapering abruptly below to form a short stipe ............ 00... e eee eee 4

Asci tapering gradually from the broadest part into an elongated stipe........... 5

4, Ascospores 27.5-30 x 4-5 um; germ slit parallel, forming a crook near the middle,

with cells readily separable only at the central septum.............. S. minima

Ascospores 37.5-40 xX 6-7.5 um; germ slit slightly to strongly oblique and cells easily

Sepatable atalleseormerits tri. Motte o.01.seiva kes Mlaa a snb arate nee ante ne aes S. australis

5. Ascospores 80-90 x 19 um; germ slits parallel to the long axis..... S. cf. megalospora

Ascospores shorter, <30 um long; germ slits parallel to oblique ................. 6

6. Germ slit parallel to oblique, with a crook near the middle............. S. isomera

Germ slit oblique, without a crook near the middle ................... S. leporina

MR Ascospores-S-Celled. 6.8. Soc. Fe Mee ha Marnis« Diem < Dncwd « Nace «Mego et ees S. pentamera

Ocospores AUST Grceleds 2). Le he's We see 2 Bash 2 ha GM 2 Ban M tte Fel taRB 2 nd S. herculea

Discussion

Due to their small size and frequent immersed habitat, Sporormiella

pseudothecia can easily be overlooked in surveys of coprophilous fungi.

Among the herbaria consulted, only two exsiccatae contained material

collected in Brazil, one (IPA37901) representing a good specimen of

S. minima and the other (URM1086) lacking a specific identification and

not well preserved enough to allow accurate revision. Our study shows

the flexibility regarding substrate preference and vegetation region by

Sporormiella species. No clear substrate and/or regional preference could be

detected. Sporormiella minima, with the highest number of records, was the

most common Sporormiella species in herbivore dung in Brazil. These fungi

usually specialize in exploiting herbivore dung and only occasionally have

been recorded on other substrates due to stresses or disturbances affecting

the community structure over the time.

Sporormiella longicolla sp. nov. (Brazil) ... 469

According to Kruys & Wedin (2009), at one time Sporormiella

circumscription was restricted to coprophilous species. Despite being

controversial, the presence of the ostiole on the ascomata (traditionally

used to separate Preussia from Sporormiella) appears to follow substrate

preferences of species for both genera: Preussia species are non-ostiolate

and found on plant debris, wood, or soil (Cain 1961), while Sporormiella

species are ostiolate and coprophilous. This statement is supported by the

polyphyly observed between the genera by Kruys & Wedin (2009) and by

the inconsistent ostiole developmental pattern observed in Sporormiaceae by

Guarro et al. (1997). The importance of active spore liberation is known in

the study of coprophilous fungi (Ingold 1965, Webster 1970, Krug et al. 2004).

However, development of the active method of liberation as an adaptation to

coprophilous substrates in Sporormiaceae has yet to be studied.

Common species, like S. minima and S. australis, were found throughout

the year alongside less common species. All exsiccatae containing

representatives of Sporormiella in Herbarium Pe. Camille Torrand (URM)

were collected in other countries under the name Sporormia and thus were

not included in this treatment.

Acknowledgments

The authors thank Profa. Marcela Caceres (Universidade Federal de Sergipe) and

Prof. Jadergudson Pereira (Universidade Estadual de Santa Cruz, Ilhéus) for expert

presubmission reviews.

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