IV ... MYCOTAXON 130(2)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY (TWO) — TABLE OF CONTENTS

COVER SECTION

PREVICW CHES 4p 2b, lade Aee deed asicck PROM ich PA ce told ob toon oA GALS Mn Meehan ic vii

1 ey ETH LD NRE Rel eG ae Sa nS A ee pe ee Slat, Aa Sh See OS eet at, RAS eS viii

BOW! Tie TE HOO. 34s 2 us Sak Ses GRO oye WES oo Bid oa ea Ka a cae CMe na ix

SUD SSIONE DRO CCU NCS x EA Ny en, Sy Mee Pe AR ER oly Se age ERS xi

RESEARCH ARTICLES

A new species of Neosporidesmium from Hainan, China

XIANG-YU LI, SHU-YAN LIU, & XIU-GUO ZHANG

A new species of Matsushimiella from submerged leaves

in the Brazilian Amazon Forest JOSIANE SANTANA MONTEIRO,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL FE. CASTANEDA-RUIZ

Amazonian phalloids: new records for Brazil and South America

TIARA S. CABRAL, CHARLES R. CLEMENT, & IURI G. BASEIA

On the generic names Kriegeria

MERJE TOOME, SHAUN R. PENNYCOOK, & M. CATHERINE AIME

New records of crustose lichens and a lichenicolous Arthonia

from Vietnam SANTOSH JOSHI, DALIP KUMAR UPRETI, SOON-OK OH,

Tut THuy NGUYEN, ANH DzuNG NGUYEN, & JAE-SEOUN HuR

First records of some Asian macromycetes in Africa PaBLo P. DANIELS,

OuMmAROU HAMA, ALFREDO JUSTO FERNANDEZ, FELIX INFANTE GARCIA-PANTALEON,

Moussa BARAGE, DAHIRATOU IBRAHIM, & MARIA Rosas ALCANTARA

Contribution to the knowledge of Inonotus baumii in Thailand

ANON AUETRAGUL, ORATAI EUATRAKOOL,

Maria LETIZIA GARGANO, & GIUSEPPE VENTURELLA

Taxonomic studies in Chrysoderma, Corneromyces,

Dendrophysellum, Hyphoradulum, and Mycobonia Karen K. NAKASONE

A new Graphilbum species from western hemlock (Tsuga heterophylla)

in Canada JAMES REID & GEORG HAUSNER

Molecular analyses reveal a new species in Melanoderma

from tropical China HaI-SHENG YUAN & YU-HE Kan

Graphis hongkongensis sp. nov. and other Graphis spp.

new to Hong Kong WEI GUO & JAE-SEOUN Hur

Two new conidial fungi from Chapada Diamantina, Brazil

CAROLINA RIBEIRO SILVA,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL F. CASTANEDA-RUIZ

307

371

315

321

329

337

361

369

399

421

429

437

APRIL-JUNE 2015... V

Anaexserticlava caatingae, a new conidial fungus from the

semi-arid Caatinga biome of Brazil TasciaNo Dos SANTOS SANTA IZABEL,

Davi AUGUSTO CARNEIRO DE ALMEIDA, JOSIANE SANTANA MONTEIRO,

Marcos FABIO OLIVEIRA MARQUES, Luis FERNANDO PASCHOLATI GUSMAO,

& RAFAEL E CASTANEDA-RUIZ 445

A new species of Endophragmiella from Guizhou, China

YING-Rut Ma, JI- WEN XIA, & XIU-GUO ZHANG 451

Peziza succosella and its ectomycorrhiza associated with Cedrus deodara

from Himalayan moist temperate forests of Pakistan

SANA JABEEN, TAYIBA ASHRAF, & ABDUL NASIR KHALID 455

Cryptocoryneum parvulum, a new species on Araucaria angustifolia

(Brazilian pine) SILVANA SANTOS DA SILVA,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL FE. CASTANEDA-RUIZ 465

Two new species and a new record of Leptogium from China

Hua-JrE Lru, MAN-QING XI, JIAN-SEN Hu, & QING-FENG WU 471

Digicatenosporium polyramosum, a new hyphomycete from Brazil

SHEILA MIRANDA LEAO-FERREIRA, Luis FERNANDO PASCHOLATI GUSMAO,

Davi AUGUSTO CARNEIRO DE ALMEIDA, & RAFAEL FE CASTANEDA-RUIZ 479

Duportella lassa sp. nov. from Northeast Asia

VIACHESLAV SPIRIN & JIRE KouT 483

Brachycephala exotica, a new hyphomycete from Brazil

JOSIANE SANTANA MONTEIRO,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL EF CASTANEDA-RUIZ 489

A new species of Diplococcium from the Brazilian semi-arid region

Davi AUGUSTO CARNEIRO DE ALMEIDA, TASCIANO Dos SANTOS SANTA IZABEL,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL F, CASTANEDA-RUIZ 495

Distophragmia, a new genus of microfungi to accommodate

Endophragmiella rigidiuscula RAFAEL FE, CASTANEDA-RUIZ,

SHEILA MIRANDA LEAO-FERREIRA, & LUIS FERNANDO PASCHOLATI GUSMAO 499

A new species of Chaetochalara on decaying leaves from Brazil

SILVANA SANTOS Da SILVA, CAROLINA RIBEIRO SILVA,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL FE. CASTANEDA-RUIZ 505

Two new species of Spadicoides and Sporidesmiella

from Yucatan, Mexico GABRIELA HEREDIA, MARCELA GAMBOA-ANGULO,

Rosa M. ARIAS, & RAFAEL F. CASTANEDA-RUIZ 511

Pholiota olivaceophylla, a forgotten name for a

common snowbank fungus, and notes on Pholiota nubigena

NOAH SIEGEL, NHU H. NGUYEN, & ELSE C. VELLINGA 517

Leucoagaricus lahorensis, a new species of L. sect. Rubrotincti

T. Qasim, T. Amir, R. Nawaz, A.R. NIAzI, & ALN. KHALID 533

v1 ... MYCOTAXON 130(2)

Aspicilia volcanica, a new saxicolous lichen from Northeast China

GULBOSTAN ISMAYIL, ABDULLA ABBAS, & SHOU-YU GUO

Redescription of Mycorrhaphium pusillum, a poorly known

hydnoid fungus Kaisa TERVONEN, VIACHESLAV SPIRIN, & PANU HALME

Micropsalliota pseudoglobocystis, a new species from China

Li WEI, YONG-HE L1, KEvIN D. HyDE, & Ru1-LIN ZHAO

Two new Rosellinia species from Southwest China

QztrRuI LI, JICHUAN KANG, & KEVIN D. HYDE

Some new records of Uredinales from Khyber Pakhtunkhwa, Pakistan

M. Fraz, A. Hasrs, N.S. AFSHAN, & A.N. KHALID

The genus Allocetraria (Parmeliaceae) in China

Ru1-FANG WANG, XIN-LI WEI, & JIANG-CHUN WEI

BOOK REVIEWS AND NOTICES ELsE C. VELLINGA (EDITOR)

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 130(2)

543

549

563

569

577

593

599

APRIL-JUNE 2015 ...

REVIEWERS — VOLUME ONE HUNDRED THIRTY (TWO)

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.

N.S. AFSHAN

JOE AMMIRATI

JULIANO M. BALTAZAR

UwE BRAUN

H.H. BuRDSALL, JR.

R. E CASTANEDA-RUIZ

Yu-CHENG Dal

KANAD Das

DENNIS E. DESJARDIN

MarTIN ESQUEDA

WALTER GAMS

ZAI-WEI GE

J. GINNS

ToM GRAFENHAN

Luis EP. GusMAOo

Lru-Fu HAN

SHUANGHUI HE

JAE-SEOUN Hur

OMOANGHE ISHIKHUEMHEN

UDENI JAYALAL

ZE-FENG JIA

SANTOSH JOSHI

BRYCE KENDRICK

S. KONDRATYUK

HEIKKI KOTIRANTA

SHAMBHU KUMAR

T. K. ARUN KUMAR

DeE-WEI LI

LAszL6 LOxK6s

THORSTEN LUMBSCH

Eric H.C. McKENZIE

Davin J. MCLAUGHLIN

GABRIEL MORENO

ABDUL REHMAN NIAZI

TUOMO NIEMELA

LORELEI L. NORVELL

CLARK L. OVREBO

OMAR PAiNno PERDOMO

SHAUN R. PENNYCOOK

DONALD H. PFISTER

ScoTT REDHEAD

Amy Y. ROSSMAN

INDREK SELL

RAGHVENDRA SINGH

JAN VONDRAK

Norou SULEIMEINE YOURO

ZE-FEN YU

EUGENE O. YURCHENKO

GEorRGIOS I. ZERVAKIS

X1u-Guo ZHANG

VII

vill ... MYCOTAXON 130(2)

ERRATA FROM PREVIOUS VOLUMES

VOLUME 123

p. 479, line 2 for: NGUYEN ANH DZUNG

VOLUME 124

p. 51, lines 2-3 for: NGUYEN ANH DZUNG

p. 309, line 2 for: NGUYEN ANH DZUNG

VOLUME 125

p. 69, line 3 for: NGUYEN ANH DZUNG

VOLUME 130(1)

p. 22, line 11 FOR: Q = 1.29-1.39

p. 61, line 9 FOR: School of Life Science

p. 61, line 13 FOR: Academia of Agriculture

read: ANH DZUNG NGUYEN

read: ANH DzZUNG NGUYEN

read: ANH DZUNG NGUYEN

READ: Q = 1.33

READ: College of Life Science

READ: Academy of Agriculture

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY (ONE)

MYCOTAXON for JANUARY-MARCH, (I-v1 + 1-306)

was issued on April 22, 2015

APRIL-JUNE 2015 ... IX

FROM THE EDITOR-IN-CHIEE

MYCOTAXON 130(2) contains 32 research papers by 112 authors (representing

18 countries) and reviewed by 50 expert reviewers. Within its pages are

four new genera (Anaexserticlava, Brachycephala, Distophragmia, and

Phaeoschizotrichum) and 26 species new to science representing Allocetraria,

Aspicilia, Endophragmiella, Graphis, Leptogium, Melanoderma, Micropsalliota,

Neosporidesmium, & Rosellinia from China; Anaexserticlava, Brachycephala,

Cryptocoryneum, Chaetochalara, —_ Digicatenosporium, —_ Diplococcium,

Distophragmia, Matsushimiella, Phaeoschizotrichum, & _ Pyriculariopsis

from Brazil; Duportella from Northeast Asia; Graphilbum from Canada;

Leucoagaricus from Pakistan; and Spadicoides & Sporidesmiella from Mexico.

In addition to range extensions (in Brazil, Niger, Pakistan, Thailand,

& Vietnam) and/or new hosts for previously named taxa, we also offer

6 new combinations (in Corneromyces, Distophragmia, Polyporus,

Pseudolagarobasidium, & Vararia), a convincing argument that Kriegeria is a

legitmate basidiomycete (not ascomycete) name, a revival of a ‘forgotten’ name

for acommon western North American pholiota, and a re-description of a rare

Mycorrhaphium.

As we noted in MycoTaxon 129(2) (the 2014 October-December issue), our

journal is temporarily without a permanent book review editor. Nonetheless,

Else Vellinga has generously returned to serve as a guest editor in this issue.

Thanks yet again, Else!

NOMENCLATURE OF AUTHORS AND AUTHORITIES— As an international journal

that publishes papers in English, MycoTAxon is not immune to nomenclatural

confusion across international borders, particularly when it comes to

distinguishing the given or generational names from the surname—that

all-important ‘family’ name that determines which of several names will be

spelled in full in abbreviated bibliographic references. We usually manage to

decipher the author names correctly, although it usually takes us one to two

months to override the stubborn Ingenta algorithm that insists on listing all

Brazilian and Portuguese names following the Spanish convention.

Asian names — in particular Chinese, Korean, and Vietnamese — present

a special challenge. For over two years, we have incorrectly displayed one

author’s name, unfortunately for Nguyen Anh Dzung (as it appears in proper

Vietnamese). Prof. Nguyen first served as co-author in MycoTAxoOn 117, where

his name appeared as Anh Dzung Nguyen (properly abbreviated and indexed

x ... MYCOTAXON 130(2)

as Nguyen AD). Unaccustomed to seeing his name in the ‘English’ style, Prof.

Nguyen contacted us to say that his name was Nguyen Anh Dzung. Although

well aware that Nguyen is a common Vietnamese surname, we did not know

whether it could also serve as a given name. Thus MycotTaxon followed the

author’s wishes by displaying his name in his next four papers in the Vietnamese

order and indexing his name as Dzung NA. With this issue, however, we

establish that Nguyen is, in fact, his surname. With past sins corrected in the

ERRATA On p. Vit, Prof. Nguyen has kindly agreed to follow our ‘English’ policy

in all future papers.

Other confusions surround how to abbreviate or decode the name of

a nomenclatural authority. MycoTAXxoNn asks its authors to adhere to the

INDEX FUNGORUM list of author abbreviations covering researchers who have

previously named fungi. This web-accessible list is handy both for authors and

for readers needing to decode the abbreviations of the authorities that stand after

the names of taxa in a full scientific name. An author proposing a new taxon for

the first time need use initials before the surname ONLY if previous authorities

have had the same surname. [I consider being known only as ‘Norvell’ both

simpler and a bit of an honor!] Those who find that their surname has already

been ‘taken’ should follow the established abbreviation (e.g., “Sm? for ‘Smith,

and add their initials. In instances where two authors share the same initials,

then one of the given names should be spelled out. Where nomenclatural

authors share all the same names, they are separated by ‘bis’ (for 2), ‘tertia’ (for

3), and so on. Be forewarned — as botanists, phycologists, and mycologists are

governed by the same code, each surname can stand alone only once. Which

explains why the late Meinhard Moser is designated by ‘M.M. Moser’ and

not simply Moser, which refers to the economic botanist Heinrich Christoph

Moser, active around 1794.

What’s in a name, indeed?

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

30 June 2015

APRIL-JUNE 2015 ... XI

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MycotTaxon publishes four quarterly issues per year. Both open access and

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- MYCOTAXON

http://dx.doi.org/10.5248/130.307

Volume 130, pp. 307-310 April-June 2015

A new species of Neosporidesmium from Hainan, China

XIANG-YU LI*?, SHU-YAN Liu?“ & XIU-GUO ZHANG? ®

' Department of Plant Pathology, Jilin Agricultural University, Changchun, 130118, China

? Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: “liussyan@163.com; *zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Neosporidesmium diaoluoshanense sp. nov., collected on rotten branches

from tropical forest of Hainan Province, China, is described and illustrated. It differs from

previously described Neosporidesmium species in producing conidia that possess a colourless,

globose, mucilaginous sheath at the apex.

KEY worps — asexual fungi, key, taxonomy

Introduction

The genus Neosporidesmium was established by Mercado & Mena (1988)

for a single species, N. maestrense. Subsequently seven other species have

been described: N. sinensis and N. microsporum (Wu & Zhuang 2005);

N. antidesmatis, N. malloti, and N. xanthophylli (Ma et al. 2011a); N. micheliae

(Zhang et al. 2011a); and N. vietnamense (Melnik & Braun 2013). The genus

is characterized by effuse, dark brown to black, hairy colonies, unbranched

conidiophores aggregating into erect, cylindrical, dark brown to black

synnemata and integrated, terminal, monoblastic, lageniform or doliiform,

determinate or percurrently proliferating conidiogenous cells that produce

solitary, pale brown to brown, obclavate or cylindrical, smooth, distoseptate

conidia. The criteria used for species delimitation are mainly based on

proliferations of conidiogenous cells, conidial morphological characteristics,

and conidiomatal overall size (Mercado & Mena 1988, Wu & Zhuang 2005).

The tropical forests of southern China have a rich mycobiota, and many

new species have been discovered there (Zhang et al. 2009a,b, 2011b, Ma et

al. 2011a,b, 2012). During a continuing investigation on saprobic microfungi

from tropical forest of Hainan Province, a morphologically distinct species of

Neosporidesmium was found. It is proposed here as a new species.

308 ... Li, Liu, & Zhang

Neosporidesmium diaoluoshanense Xiang Y. Li & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 812118

Differs from other Neosporidesmium species in its conidia having an apical, colourless,

globose, mucilaginous sheath.

Type: China, Hainan Province: Diaoluoshan Nature Reserve, on rotten branches of an

unidentified tree, 10 Apr. 2014, H.X. Dong (Holotype HSAUP H7562; isotype, HMAS

243460).

EryMo_oey: in reference to the type locality.

Co.ontgs on the natural substrate effuse, solitary, dark brown, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of dark brown

to black, septate, cylindrical hyphae. ConIDIOMATA synnemata, indeterminate,

scattered, erect, up to 605 um high, 40-60 um wide, composed of straight,

unbranched, septate, smooth, brown to dark brown conidiophores 3-6 um

wide, diverging terminally and laterally. CoNIDIOGENOUS CELLS monoblastic,

integrated, determinate, smooth, cylindrical, doliiform or lageniform, brown to

dark brown, 4-10 x 9-15 um. Conidial secession schizolytic. Conrp1a solitary,

acrogenous, obclavate, base truncate, 8-11-distoseptate, 105-135 x 10.5-13.5

um, 2.5-3.5 um wide at the truncate base, rounded at the apex, dry, smooth-

walled, pale brown, but hyaline at the apical cells, with an apical, globose,

hyaline, 10.5-13.5 um diam., mucilaginous sheath.

ComMENTsS - Neosporidesmium diaoluoshanense differs from all previously

described species in the genus by producing conidia with an apical, globose,

mucilaginous, hyaline sheath and having scattered synnemata. The new species

resembles N. micheliae, which differs by having conidia with fewer septa (7-8)

and lacking a mucilaginous sheath.

Key to Neosporidesmium species

l.. Conidiogenous cells-with percurrent prolifetation: sc .::.32sess eee dee oe 2

Conidiogenous cells without percurrent proliferation ...................04. 4

2h. Wy Cotidiamnearlyoylindricale Wis siot sano ttle ashen busethae Restathoo ack N. microsporum

@onidia.obelavate 2.5. bm the Ame ke me Meee Am a GB head 3

3.. gC onidiasvwuth- filiform beak. 90.9 7 IR Ee PS N. antidesmatis

GConidiawithoutibedk ss dis... a dete «a dinanecd diaeeg Db peed baer d Dhoede N. maestrense

4. Syritietmata Scattered s Peete dh sabes deh sukea dye pikes dr gpeteg dye peda Ae poder fe seb d inp ckm repeal 5

Syimematanouscatteted At. turk t, ataug ta Aue toh oe ee OY oe eRe one oan. te ae 6

Daw P@Onidia-distoseptate teehee shin gl igh adie dog ad oy N. diaoluoshanense

COnidig"eusepiales ei. 5 -Seis xp Hels xa -dlelnge oleae lelage® lett Steiner 4s N. micheliae

6 Gonidrawith beakig . b..idine+ddike edd caegd naegd bard weedd bow dibapews Qarvuls 7

COMATARWVIENOUL DEA Hert eA forte) Atel A Sat lB ah lah ic Bat uleSat 5 Cae N. malloti

Fite, PROT TAR CUSE IAL ES acter ote schire e chce lik a ee a mR N. vietnamense

omIIaPIStOSE PALE! Soups droog tr ponses drinsceg dr pee dh preg aye og ayn gedoe ct peta ea gctoe 8

Neosporidesmium diaoluoshanense sp. nov. (China) ... 309

A | B

20 um

Fic. 1. Neosporidesmium diaoluoshanense. A. Synnemata on natural substratum.

B. Conidia with an apical, hyaline, globose, mucilaginous sheath. C, D. Synnemata

with conidiophores and conidia.

&. ) sConidia with: 6-10 distoséptany. ams ocaniettiaptcly analy ata N. xanthophylli

Coridia svat 1O=TLdistosepta soa ipa earnbryoaeetetoa peter te N. sinensis

Acknowledgments

The authors express gratitude to Dr. Eric H.C. McKenzie and Dr. R.F Castafieda

Ruiz for serving as pre-submission reviewers and for their valuable comments and

suggestions. This project was supported by the National Natural Science Foundation of

310... Li, Liu, & Zhang

China (Nos. 31093440, 31230001) and the Ministry of Science and Technology of the

People’s Republic of China (Nos. 2006FY120100).

Literature cited

Ma J, Wang Y, Ma LG, Zhang YD, Castafeda-Ruiz RF, Zhang XG. 2011a. Three new species of

Neosporidesmium from Hainan. Mycol. Progress 10: 157-162.

http://dx.doi.org/10.1007/s11557-010-0685-2

Ma J, Wang Y, O’Neill NR, Zhang XG. 2011b. A revision of the genus Lomaantha, with the

description of a new species. Mycologia 103(2): 407-410.

http://dx.doi.org/10.1007/s11557-010-0696-z

Ma J, Zhang YD, Ma LG, Ren SC, Castafeda-Ruiz RF, Zhang XG. 2012. Three new species of

Solicorynespora from Hainan, China. Mycol. Progress 11: 639-645.

http://dx.doi.org/10.3852/ 10-176

Mercado Sierra A, Mena Portales J. 1988. Nuevos o raros hifomicetes de Cuba. Acta Bot. Cubana

59: 1-6.

Mel'nik VA, Braun U. 2013. Atractilina alinae sp. nov. and Neosporidesmium vietnamense sp. nov. -

two new synnematous hyphomycetes from Vietnam. Mycobiota 3: 1-9.

http://dx.doi.org/10.12664/mycobiota.2013.03.01

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Diversity Research Series 15. 351 p.

Zhang K, Ma J, Wang Y, Zhang XG. 2009a. Three new species of Piricaudiopsis from southern

China. Mycologia 101(3): 417-422. http://dx.doi.org/10.3852/08-147

Zhang K, Ma LG, Zhang XG. 2009b. New species and records of Shrungabeeja from southern

China. Mycologia 101(4): 573-578. http://dx.doi.org/10.3852/09-006

Zhang YD, Ma J, Ma LG, Castafieda-Ruiz R.F, Zhang XG. 2011a. New species of Phaeodactylidium

and Neosporidesmium from China. Sydowia 63: 125-130.

Zhang YD, Ma J, Wang Y, Ma LG, Castafeda-Ruiz R.E, Zhang XG. 2011b. New species and

record of Pseudoacrodictys from southern China. Mycol. Progress 10: 261-265. http://dx.doi.

org/10.1007/s11557-010-0696-z

MY COTAXON

http://dx.doi.org/10.5248/130.311

Volume 130, pp. 311-314 April-June 2015

A new species of Matsushimiella from submerged leaves

in the Brazilian Amazon Forest

JOSIANE SANTANA MONTEIRO’, LuiS FERNANDO PASCHOLATI GUSMAO”™,

& RAFAEL E CASTANEDA-RUIZ?

‘Departamento de Ciéncias Biologicas, Laboratorio de Micologia, Universidade Estadual

de Feira de Santana, BR116 KM03, 44031-460, Feira de Santana, Brazil

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’

(INIFAT), Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: lgusmao@uefs. br

AxBsTRACT—Matsushimiella paraensis sp. nov. is described and illustrated from specimens

collected on submerged leaves in a stream, from Para, Brazil, in the Amazon rainforest.

The fungus is characterized by polyblastic sympodially extending conidiogenous cells and

ellipsoid, long ovoid to broadly oblong, 2-4-distoseptate medium to golden brown smooth

conidia with a pale brown to subhyaline basal frill left by rhexolytic conidial secession.

KEY worDs— taxonomy, freshwater fungi, tropics

Introduction

During a mycological survey of conidial fungi occurring on submerged

decaying plant remnants from the Amazon rainforest in Brazil (Monteiro

et al 2014a,b), an interesting fungus was collected on submerged leaves. Its

conidiogenesis and conidial features clearly suggest a placement in the hitherto

monotypic genus Matsushimiella R.F. Castafieda & Heredia (Castaneda-Ruiz

et al. 2001). However, there are remarkable conidial differences distinguishing

Matsushimiella queenslandica (Matsush.) R.E. Castafieda & Heredia, the type

species of the genus, and the new collections from Brazil.

Material & methods

Samples of submerged litter were placed in paper and plastic bags, taken to the

laboratory, and treated according to Castafieda-Ruiz (2005). Mounts were prepared in

PVL (polyvinyl alcohol, lactic acid, and phenol) and measurements were taken at x1000.

312 ... Monteiro, Gusmao, & Castafieda-Ruiz

Micrographs were obtained with an Olympus microscope (model BX51) equipped with

bright field and Nomarski interference optics. The holotype and an additional specimen

are deposited in the Herbarium of Universidade Estadual de Feira de Santana, Brazil

(HUEEFS).

Taxonomy

Matsushimiella paraensis J.S. Monteiro, R.F. Castafieda & Gusmao, sp. nov. Fia.1

MycoBank MB 807635

Differs from Matsushimiella queenslandica by its larger, ellipsoid, long ovoid to broad

oblong, 2-4-distoseptate conidia.

Type: Brazil, Para State, Belém, Parque Estadual do Utinga, 1°25’S 48°27’W, on

submerged leaves of an unidentified plant in a stream, 9 Jan. 2013, coll. J.S. Monteiro

(Holotype: HUEFS 210425).

EryMo_oey: Latin paraensis refers to Para State, where the type specimen was collected.

CoLonies on the natural substrate effuse, hairy, smooth, golden brown.

CONIDIOPHORES distinct, unbranched, erect, cylindrical below, geniculate

above, 5-10-septate, smooth, wall somewhat thickened, up to 1.5 um wide, dark

brown at the base, pale brown toward the apex, smooth, 170-220 x 5-8 um.

CONIDIOGENOUS CELLS integrated, terminal, geniculate, with several sympodial

extensions, polyblastic, pale brown, 40-70 x 4-5 um. SEPARATING CELLS more

or less cylindrical, subhyaline, 3-4 x 2-2.5 um, fracturing and remaining

as a persistent short peg. Conidial secession rhexolytic. Conip1a solitary,

acropleurogenous, ellipsoid, long ovoid to broadly oblong, 2-4-distoseptate,

mostly 4-distoseptate, 20-25 x 8-10 um, wall 1-1.5 um thick, smooth, mid

brown or golden brown, fimbriate at the base, frill subhyaline to pale brown,

1-2 um long, 2-2.5 um wide.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, ParA, Parque Estadual do Utinga, 1°25’S

48°27’W, on submerged leaves of an unidentified plant in a stream, 17 Oct. 2012, coll.

J.S. Monteiro (HUEFS 210427).

Note: Matsushimiella is typified by M. queenslandica (Castafieda-Ruiz et al.

2001) based on Pseudospiropes queenslandica Matsush., which was collected

from decaying leaves of an unidentified plant in Australia. The existence of the

separating cells was mentioned in the brief discussion by Matsushima (1989),

but not described in the original description of Matsushimiella (Castafeda-

Ruiz et al. 2001). Seifert et al. (2011) characterized the conidiogenous cells

as polytretic, which refers to the original description of Matsushimiella

(Castafieda-Ruiz et al. 2001). However, conidiogenesis is holoblastic, with

mostly polyblastic conidiogenous cells. Matsushimiella queenslandica is readily

distinguishable from the new Brazilian species by its obovate, 1-3-, mostly

2-distoseptate, 9-16 x 6-8 um, smooth, pale brown conidia that secede by

rhexolytic fracture of cylindrical separating cells.

Matsushimiella paraensis sp. nov. (Brazil) ... 313

Fic. 1. Matsushimiella paraensis (holotype HUEFS 210425): A. Conidia. B. Conidiophore,

conidiogenous cell, separating cells, and conidia. C. Conidiogenous cells. D-F. Conidiogenous

cells, separating cells (arrow), and conidia. Scale bars = 10 um.

314 ... Monteiro, Gusmao, & Castafieda-Ruiz

Acknowledgments

The authors express their sincere gratitude to Uwe Braun and Eric H.C. McKenzie

for their critical reviews of the manuscript. The authors are grateful to the Coordination

for the Improvement of Higher Education Personnel (CAPES) for financial support and

the ‘Programa de Pés-graduacao em Biologia de Fungos - PPGBF/UFPE: The authors

thank the support provided by ‘Programa Ciéncia sem Fronteiras’ RFCR is grateful

to Cuban Ministry of Agriculture and ‘Programa de Salud Animal y Vegetal, project

P131LH003033 for facilities. We acknowledge the assistance provided by Paul M. Kirk

and Vincent Robert and Arthur de Cock through the IndexFungorum and Mycobank

websites. Lorelei Norvell’s editorial and Shaun Pennycook’s nomenclatural reviews are

greatly appreciated.

Literature cited

Castaneda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. Anais do V

Congresso Latino Americano de Micologia, Brasilia: 182-183.

Castafieda-Ruiz RF, Heredia G, Reyes M, Arias RM, Decock C. 2001. A revision of the genus

Pseudospiropes and some new taxa. Cryptogamie Mycologie 22: 3-18.

http://dx.doi.org/10.1016/S0181-1584(01)01057-0

Matsushima T. 1989. Matsushima mycological memoirs no. 6. Matsushima Fungus Collection,

Kobe, Japan.

Monteiro JS, Gusmao LFP, Castafieda-Ruiz RF. 2014a. Helicodochium, a new microfungus from

submerged wood in Brazil. Mycotaxon 127: 5-9. http://dx.doi.org/10.5248/127.5

Monteiro JS, Gusmao LFP, Castafeda-Ruiz RE. 2014b. Two new microfungi from Brazilian Amazon

Forest: Atrogeniculata submersa and Nigrolentilocus amazonicus. Mycotaxon 127: 39-45.

http://dx.doi.org/10.5248/127.39

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9: 1-997. http://dx.doi.org/10.3767/003158511X617435

MYCOTAXON

http://dx.doi.org/10.5248/130.315

Volume 130, pp. 315-320 April-June 2015

Amazonian phalloids:

new records for Brazil and South America

TIARA S. CABRAL', CHARLES R. CLEMENT’, & IURI G. BASEIA3

‘Programa de Pés-graduacdo em Genética, Conservacdo e Biologia Evolutiva &

?Coordenacao de Tecnologia e Inovacao, Instituto Nacional de Pesquisas da Amazonia,

Av. André Araujo, 2936 - Petropolis, Manaus, Amazonas, 69067-375, Brazil

*Departamento de Botanica e Zoologia, Universidade Federal do Rio Grande do Norte,

Natal, Rio Grande do Norte, 59072-970, Brazil

* CORRESPONDENCE TO: ttiara@gmail.com

ABSTRACT — Recent field trips in the Amazonian rainforest revealed two uncommon

phalloid species, Lysurus arachnoideus (new for Brazil) and Phallus cinnabarinus (new for

South America). Detailed morphological descriptions, photos, and taxonomic remarks are

presented.

KEY worps — gasteroid fungi, Lysuraceae, Neotropics, new records, Phallaceae

Introduction

Corda (1842) proposed the family Lysuraceae (as “Lysuroidea”) when he

segregated the genera Lysurus Fr. and Aseroe Labill. from Clathraceae. Though

many authors did not accept Lysuraceae (Cunningham 1944, Zeller 1949,

Dring 1980), Hosaka et al. (2006) showed that Lysuraceae is phylogenetically

more closely related to Phallaceae than to Clathraceae, and considered it

an independent family, comprising the single genus Lysurus Fr., originally

proposed to accommodate L. mokusin (Fries 1823).

Kirk et al. (2008) accept five species in Lysurus, although Index Fungorum

lists 41 names for the genus. Diagnostic morphological features for Lysurus

include a long stipe with a distinct fertile portion consisting of arms or vertical

columns that are united or free or that form a network and a gleba lying on

the inner surface of each arm or column (Dring 1980, Trierveiler-Pereira et al.

2014).

The genus Phallus Junius ex L. includes species recognized by their phallic

shape that comprises an immature basidioma enclosed in a volva that breaks

316 ... Cabral, Clement, & Baseia

with maturity and from which emerges a pseudostipe with a fetid mucilaginous

gleba. Phallus species are widely distributed, but Kreisel (1996) indicated that

the center of diversity is China and Southeast Asia.

Thirteen Phallus records and five Lysurus species are currently recognized

from Brazil (Trierveiler-Pereira & Baseia 2009, Cortez et al. 2011); some

doubtful taxa have also been reported. This paper contributes to the taxonomic

knowledge of phalloid diversity in Amazonia.

Materials & methods

The specimens were collected during field trips in Brazilian Amazonian rainforests

(Careiro, state of Amazonas, Brazil) and French Guiana (Cayenne), where ecological

notes and photographs were taken. Morphological measurements were taken from

fresh and dried specimens according to Dring (1980) and Cortez et al. (2011). Color

descriptions follow Kueppers (1982). Microstructures were measured after rehydration

in KOH 5%, with 20 measurements made randomly for each structure using a Leica

DM 2500 microscope. Spore and hyphal images were captured with a coupled EC3

camera and Leica Application suite v.2.1.0 software. The specimens were deposited

at the Instituto Nacional de Pesquisas da Amazénia Herbarium, Manaus, Amazonas,

Brazil (INPA).

Taxonomy

Lysurus arachnoideus (E. Fisch.) Trierv.-Per. & Hosaka, Mycologia

106: 909, 2014. FIG. 1

= Aseroe arachnoidea E. Fisch., Denkschr. Schweiz. Naturf.

Ges. WS. Lee, Mycologia 32(1): 76, 1890.

Eces 15-20 mm diam., globose to subglobose, pale yellow (NOOY20MO00),

semi-hypogeous, thin rhizomorphs present. EXPANDED BASIDIOMATA 40 x 11

mm. Volva 27-21 mm wide, white (A00M00C00) to pale yellow (NOOY20M00),

outer layer composed of filamentous hyphae 4.6-6.4 um wide, septate with

clamp connections, inner layer gelatinous, composed of 4-5.5 um wide

filamentous hyphae. Pseudostipe 12 x 11 mm, cylindrical, white (AO0M00C00);

surface rugose, hollow, ending in a central perforated disc; composed of globose

to subglobose pseudoparenchymatous hyphae, 70 um diam., hyaline in 5%

KOH. Receptacle formed by a disc with 9 arms developing from the margins.

Arms up to 18 mm long, not cylindrical, hollow and consisting of a single tube,

white (AO0M00C00), inner surface rugose, covered by the gleba up to 2/3 of the

arm length. Gleba olive brown (N99A50M10), fetid. Bastp1osPpoREs 3.5-4.4 x

2.1-2.5 um, ellipsoid, smooth, greenish in 5% KOH.

ECOLOGY & DISTRIBUTION: Hosaka (2010) reports L. arachnoideus from

Africa, Thailand, Japan, China, Indonesia, Sri Lanka, and Malaysia.

SPECIMENS EXAMINED — BRAZIL. AMAZONAsS: Careiro, Purupuru community,

3°22.96’S 59°42.63’W, in dung, in an ombrophilous dense forest fragment, 7.112013, TS.

Phalloids new for Brazil ... 317

20 mm

Figure 1. Lysurus arachnoideus (INPA 256537). A: Expanded basidioma. B: Basidiospores.

Cabral 41 (INPA 256537; Genbank KJ764820). JAPAN. TorTort: Kokoje, 1.[X.1980, T.

Hongo & E. Nagasawa (TMI 6927); 5.1X.1980, E. Nagasawa (TMI 6929); 9.1X.1980, E.

Nagasawa (TMI 6930); 22.1X.1983, I. Arita (TMI 11622).

ComMMENTS — Lysurus arachnoideus sensu lato was previously recorded

only once from South America, by Fischer (1928), who described a new

variety (Aseroe arachnoidea var. americana) based on a specimen collected

in Suriname, which apparently differed from A. arachnoidea var. arachnoidea

mainly by the arms being arranged in pairs. Unfortunately Fischer (1928)

did not provide a detailed description or include microstructure sizes. Our

specimen has unpaired arms (Fic. 1A), and the spore size matches an African

specimen described by Dring (1964). The taxonomically significant characters

do not vary significantly between the Brazilian and the Japanese specimens.

The Japanese material has a pseudostipe and arms longer than the Brazilian

specimen, pseudoparenchymatous hyphae that are up to 76 um diam., and

smooth spores that are up to 4 um long. Despite these differences, we prefer

to maintain our collection as L. arachnoideus (instead of A. arachnoidea var.

americana E. Fisch.) based on the morphological characters until further

molecular analyses can be carried out.

Our Amazonas specimen represents the second record for South America

and the first for Brazil.

Phallus cinnabarinus (WS. Lee) Kreisel, Czech Mycol. 48: 278, 1996. Fic. 2

= Dictyophora cinnabarina WS. Lee, Mycologia 49: 156, 1957.

EGcs 24-23 mm wide, ovoid to subglobose, pale yellow (N10A10MO00) to

brown (N60A90M00), with thick and lilac to pink rhizomorphs. EXPANDED

BASIDIOMATA 192 mm high. Receptacle 34 high x 70 mm wide, bright

orange (N10A70M70), campanulate, with an apical pore, surface reticulate.

318 ... Cabral, Clement, & Baseia

Figure 2. Phallus cinnabarinus (INPA 255835). A: Expanded basidioma. B: Basidiospores.

C: Pseudoparenchymatous hyphae of pseudostipe with pinkish pigment droplets.

Pseudostipe 130 x 23 mm, cylindrical, spongy, pale pink (N10A40M30),

composed of globose to subglobose pseudoparenchymatous hyphae, 40-67 um

diam., hyaline in 5% KOH, pinkish pigment droplets present. Volva pale yellow

(N10A10MO00), outer layer cottony, composed of filamentous hyphae 3-4 um

diam., sinuous, septate, clamp connection present; inner layer gelatinous,

composed of filamentous hyphae 2.2-3 um diam., hyaline in KOH 5%; basal

portion with rhizomorphs. Indusium extending to half of pseudostipe, bright

orange (N10A80M50), 69 mm long, polygonal meshes up to 9 mm, formed

of globose to subglobose hyphae, 24-66 um diam., pinkish pigment droplets

present. Gleba brown (N60A90M40), mucilaginous. BASIDIOSPORES 2.8-4.2 x

1.2-1.9 um, ellipsoid, smooth, olive brown in 5% KOH.

ECOLOGY & DISTRIBUTION — Kreisel (1996) first described Phallus

cinnabarinus from Taiwan (Kreisel 1996), but it has since been reported from

many parts of the world (Hemmes & Desjardin 2009).

SPECIMENS EXAMINED — BRAZIL, PaRA: Belém, Jardim Botanico Bosque Rodrigues

Alves, 1°25.83’S 48°27.30’W, on soil among grasses and at the base of bamboo clumps,

27.X1.2013, T.S. Cabral 81 (INPA 255835; Genbank KJ764821); Museu Paraense Emilio

Phalloids new for Brazil ... 319

Goeldi, 1°27.55’S 48°28.60’W, 28.X1.2013, T.S. Cabral 82 (INPA 255836). FRENCH

GUIANA: Cayenne, in grounds of the Herbier de Guyane (CAY), Institut de Recherche

pour le Developpement (IRD), 4°56.35’N 52°17.21’W, 11.III.2013, Cabral TS 48 (INPA

255837).

CoMMENTS — ‘There is controversy involving two morphologically similar

Phallus species: P. multicolor (Berk. & Broome) Cooke and P. cinnabarinus.

Phallus multicolor [= Dictyophora multicolor Berk. & Broome] was originally

described as having a lemon-yellow indusium, an orange receptacle, and

spores up to 5 um (Berkeley & Broome 1883), while P cinnabarinus was

described with a cinnabar-red receptacle and indusium, volva with pinkish

rhizomorphs at base, and spores up to 4 um (Lee 1957). Considering the

original descriptions, we believe that the specimens analysed here represent

P. cinnabarinus, based mainly on the cinnabar-red color of the indusium and

receptacle, spores smaller than in P multicolor, and divergence in rhizomorph

colours. Cunningham (1944) described an Australian specimen of Phallus

multicolor, mentioning a salmon-pink indusium, orange receptacle, volva with

short soft spines, and basidiospores up to 4.5 um long. Reporting P multicolor

from India, Dutta et al. (2012) cited a lemon-yellow indusium, lemon-yellow to

yellowish orange receptacle, and different basidiospore size. Although Hemmes

& Desjardin (2009) provide good photos of both species that illustrate the

primary macroscopic differences (indusium and receptacle colours) between

the two species, unfortunately, a detailed description of P. multicolor cannot be

found in the recent literature.

There is no previous record of P. cinnabarinus for South America. Cheype

(2010) reported P. aff. multicolor from French Guiana, but he does not

mention a herbarium voucher, which prevents further comparative analysis.

A Phallus specimen collected on a recent field trip to French Guiana enabled a

comparative study that confirmed the conspecificity of the French Guianan and

Brazilian collections as P cinnabarinus.

Acknowledgments

This study was supported by the Brazilian funding agencies CNPq (473422/2012-

3) and FAPEAM (3137/2012). The authors thank Herbarium CAY, Piero Giuseppe

Delprete, and Rémi Girault for support during the field trip in Cayenne. The authors

also thank Eiji Nagasawa for cordially lending materials from the Herbarium TMI, and

Clark Ovrebo and Martin Esqueda for reviewing the manuscript.

Literature cited

Berkeley MJ, Broome CE. 1883. List of fungi from Brisbane, Queensland; with descriptions of new

species. Part II. Transactions of the Linnean Society of London, Botany, 2nd Series, 2: 53-73.

Cheype JL. 2010. Phallaceae et Clathrus récoltés en Guyane Francaise. Bulletin Mycologique et

Botanique Dauphiné-Savoie 66: 51-66.

Corda ACJ. 1842. Icones fungorum hucusque cognitorum. 5: 1-92.

320 ... Cabral, Clement, & Baseia

Cortez VG, Baseia IG, da Silveira RMB. 2011. Two noteworthy Phallus from southern Brazil.

Mycoscience 52: 436-438. http://dx.doi.org/10.1007/S10267-011-0124-5

Cunningham GH. 1944. The gasteromycetes of Australia and New Zealand. J. McIndoe. 236 p.

Dring DM. 1964. Gasteromycetes of west tropical Africa. Mycological Papers 98. 60 p.

Dring DM. 1980. Contributions towards a rational arrangement of the Clathraceae. Kew Bulletin

35: 1-96. http://dx.doi.org/10.2307/4117008

Dutta AK, Chakraborty N, Pradhan P, Acharya K. 2012. Phallales of West Bengal, India. II.

Phallaceae: Phallus and Mutinus Arun. Researcher 4: 21-25.

Fischer E. 1928. Untersuchungen tiber Phalloideen aus Surinam. Vierteljahrsschrift der

Naturforschenden Gesellschaft in Zitirich, 73(Beibl. 15): 1-39.

Fries EM. 1823. Systema Mycologicum. 2: 276-620.

Hemmes DE, Desjardin DE. 2009. Stinkhorns of the Hawaiian Islands. Fungi 2: 8-10.

Hosaka K, Bates ST, Beever RE, Castellano MA, Colgan III W, Dominguez LS, Nouhra ER, Gem J,

Giachini AJ, Kenney SR, Simpson NB, Spatafora JW, Trappe JM. 2006. Molecular phylogenetics

of the gomphoid-phalloid fungi with an establishment of the new subclass Phallomycetidae and

two new orders. Mycologia 98: 949-959.

Hosaka K. 2010. Preliminary list of Phallales (Phallomycetidae, Basidiomycota) in Taiwan. Memoirs

of the National Science Museum, Tokyo 46: 57-64.

Kirk PM, Cannon PF, David JC, Stalpers JA. 2008. Ainsworth and Bisby’s Dictionary of the Fungi.

10th ed. Surrey: CABI/International Mycological Institute.

Kreisel H. 1996. A preliminary survey of the genus Phallus sensu lato. Czech Mycology 48: 273-281.

Kueppers H. 1982. Color atlas. Barrons Educational Series, NY.

Lee WS. 1957. Two new phalloids from Taiwan. Mycologia 49: 156-158.

http://dx.doi.org/10.2307/3755742

Trierveiler-Pereira L, Baseia IG. 2009. A checklist of the Brazilian gasteroid fungi (Basidiomycota).

Mycotaxon 108: 441-444. http://dx.doi.org/10.5248/108.441

Trierveiler-Pereira L, Silveira RMB da, Hosaka K. 2014. Multigene phylogeny of the Phallales

(Phallomycetidae, Agaricomycetes) focusing on some previously unrepresented genera.

Mycologia 106: 904-911. http://dx.doi.org/10.3852/13-188

Zeller SM. 1949. Keys to the orders, families and genera of the gasteromycetes. Mycologia 41(1):

36-58.

MYCOTAXON

http://dx.doi.org/10.5248/130.321

Volume 130, pp. 321-328 April-June 2015

On the generic names Kriegeria

MERJE TOOME’, SHAUN R. PENNYCOOK? & M. CATHERINE AIME‘*

' Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN 47907, USA

? Landcare Research, Private Bag 92170, Auckland 1072, New Zealand

*CORRESPONDENCE TO: maime@purdue.edu

AsBsTRACT — The basidiomycetous genus Kriegeria Bres. 1891 (Kriegeriaceae), typified by

K. eriophori, is legitimate. An ascomycetous genus “Kriegeria” was originally published

by Rabenhorst in 1876 as an invalid provisional name based on Ombrophila kriegeriana.

Subsequently, the ascomycetous genus was validly published as Kriegeria Hohn. 1914

(Rutstroemiaceae), typified by Peziza elatina; this genus is an illegitimate later homonym,

currently treated as a synonym of Rutstroemia. Additionally, Rabenhorst’s invalid name

was “revived” (validated) as Kriegeria Rabenh. ex Seaver 1943, typified by O. kriegeriana, to

accommodate species currently placed in Chloroscypha. Revised nomenclators are presented

for both of the genera, as well as for Ombrophila kriegeriana.

Key worps — Helotiales, Kriegeriales, Leotiomycetes, Microbotryomycetes, Xenogloea,

Zymoxenogloea

Introduction

Two different genera have been named as Kriegeria. The citation of one of

these, the basidiomycetous genus Kriegeria published by Bresadola (1891),

is uncontroversial. However, the correct citation of the ascomycetous genus

Kriegeria has been the subject of long-standing debate, attributed either

to Rabenhorst (1876, 1878a) or to Hohnel (1914); and consequently, the

basidiomycetous Kriegeria Bres. has been considered to be either an illegitimate

later homonym, or a legitimate earlier homonym. ‘The basis of this controversy

is whether Kriegeria Rabenh. is a validly published alternative name, or an

invalid provisional name. We have conducted an extensive literature survey

and nomenclatural analysis to resolve this problem.

Kriegeria Hohn. (Rutstroemiaceae, Helotiales, Leotiomycetes)

Kriegeria Hoéhn., Sitzungsber. K. Akad. Wiss., Math.-Naturwiss. Kl, Abt. 1, 123: 114.

1914, nom. illeg., non Bres. 1891.

TyPE: Peziza elatina Alb. & Schwein. : Fr., Consp. Fung. Lusat.: 330. 1805.

322 ... Toome, Pennycook, & Aime

= Rutstroemia P. Karst., Bidr. Kann. Finl. Nat. Folk 19: 12. 1871.

Tye: Peziza firma Pers. : Fr., Syn. Meth. Fung.: 658. 1801.

= Kriegeria Rabenh. ex Seaver, Mycologia 35: 492. 1943,

nom. illeg., non Bres. 1891, nec Hohn. 1914.

[“Kriegeria” Rabenh., Fungi Europ. Exsicc. 24: no. 2315. 1876, nom. prov.]

Type: Ombrophila kriegeriana Rabenh., Fungi Europ. Exsicc. 24: no. 2315. 1876.

Ombrophila kriegeriana Rabenh., Fungi Europ. Exsicc. 24: no. 2315. 1876.

[“Kriegeria olivacea” Rabenh., Fungi Europ. Exsicc. 24: no. 2315. 1876, nom. prov.]

= Ciboria kriegeriana (Rabenh.) Rehm, Hedwigia 22: 36. 1883.

= Chlorosplenium kriegerianum (Rabenh.) Sacc., Syll. Fung. 8: 318. 1889.

= Kriegeria kriegeriana (Rabenh.) Seaver, Mycologia 35: 493. 1943.

= Peziza elatina Alb. & Schwein. : Fr., Consp. Fung. Lusat.: 330. 1805.

= Helotium elatinum (Alb. & Schwein. : Fr.) Quél., Ench. Fung.: 309. 1886.

= Chlorosplenium elatinum (Alb. & Schwein. : Fr.) Sacc., Syll. Fung. 8: 318. 1889.

= Rutstroemia elatina (Alb. & Schwein. : Fr.) Rehm, Rabenh. Krypt.-Fl.,

Ed. 2, 1(3): 767. 1893.

= Kriegeria elatina (Alb. & Schwein. : Fr.) Héhn., Sitzungsber. K.

Akad. Wiss., Math.-Naturwiss. KI., Abt. 1, 123: 114. 1914.

The critical publication concerning the ascomycetous Kriegeria is the protologue

of Ombrophila kriegeriana (Rabenhorst 1876). This printed exsiccata label

presents a Latin description, Latin collection data [K6nigstein, Saxony, on

Abies twigs], a paragraph of German “Obs.’, and the author's initials “L.R.” In

the Observations, Rabenhorst noted: (1) that the specimens are immature,

creating some doubt whether Ombrophila is the appropriate genus [indicated

by inclusion of a question mark after the genus name in the binomial proposal];

(2) that the collector, W. Krieger, will endeavor to collect more material with

mature ascomata; (3) that the species may require the proposal of a new genus

for which he would suggest the name Kriegeria; (4) that in this potential new

genus, he would suggest the epithet olivacea. He did not combine the genus

and epithet into an explicit binomial (Seaver 1943 appears to be the first author

to assemble the binomial). Considered as a whole, the Observations do not

suggest that “Kriegeria olivacea” was being formally proposed as an alternative

name for Ombrophila kriegeriana, but that it was merely being suggested as a

provisional name to be taken up in the future if mature specimens justified the

proposal of a new genus. “Kriegeria” Rabenh. and “Kriegeria olivacea” Rabenh.

are therefore invalid names (McNeill et al. 2012: Art. 36.1(b)).

This situation is somewhat obscured in Rabenhorst (1878a), where the

Ombrophila kriegeriana protologue is reproduced verbatim, but with the

first two sentences of the Observations omitted; ie., there is no mention of

the immaturity of the specimens nor of Krieger's intention to look for mature

material. (Nevertheless, when Donk (1958) reached his firm decision that

Kriegeria Bres. vs. Kriegeria Hohn. & Kriegeria Rabenh. ... 323

“Kriegeria” Rabenh. was a provisional name, it was this reference that he

cited and its abbreviated version of the original Observations that he quoted.)

Because Rabenhorst (1878a) has been widely (but erroneously) accepted

as the protologue of O. kriegeriana/“K. olivacea’, several mycologists have

misunderstood “Kriegeria olivacea” to have been proposed as a valid alternative

name. It is significant that the Index of Names for Hedwigia vol. 17 (Rabenhorst

1878b: p. V) does not list either “Kriegeria” or “Kriegeria olivacea”. Rabenhorst

died on 24 April 1881, presumably before Krieger had collected or sent any

mature specimens.

Winter (1881: 70-71) commented on Ombrophila kriegeriana, reporting that

he had often seen it during spring in Zurich. On the basis of his own observations

and Rabenhorst’s description, Winter concluded that O. kriegeriana was a

synonym of Peziza elatina Alb. & Schwein.; he made no mention of the names

“Kriegeria” and “Kriegeria olivacea”.

Ombrophila kriegeriana was recombined in Ciboria by Rehm (1883: 36-37)

and in Chlorosplenium by Saccardo (1889: 318). Both authors mentioned

Winter's proposed synonymy with Peziza elatina (without accepting it), but

neither of them mentioned the names “Kriegeria” and “Kriegeria olivacea”.

Rehm (1893: 767) recombined Peziza elatina in Rutstroemia (the genus

in which this species is currently accepted). Rehm’s R. elatina nomenclator

included Ombrophila kriegeriana and its Ciboria and Chlorosplenium

recombinations as synonyms, but it did not include “Kriegeria olivacea” (even

though Rabenhorst’s suggestion of this genus and this epithet are annotated in

Rehm’s synopsis of the literature).

Hohnel (1914: 112-114) was the first author to validate the ascomycetous

genus Kriegeria, but his treatment is full of errors:

(1) Hohnel attributed the genus to “Winter (Hedwigia, 1878, 17. Bd., p.

32)

(2) he stated that Peziza elatina was transferred to Kriegeria by “Winter

(Hedwigia, 1878, 17. Bd., p. 32)”; and

(3) he stated that the generic type was “Kriegeria elatina (A. u. S.) Winter”.

The cited Hedwigia reference has no connection with Winter and is, in fact,

Rabenhorst (1878a), which contains no mention of Kriegeria elatina or its

basionym Peziza elatina. Rabenhorst edited the first ten issues of Hedwigia

vol. 17 (pp. 1-160, January-October 1878), and Winter's editorship did not

commence until issue 11 (November 1878). (Likewise, Winter did not assume

editorship of Fungi Europaei Exsiccati until late 1881, after Rabenhorst’s death.)

We have scrutinized all of Winter’s publications from 1878 until his death

in 1887 and can find no mention of either “Kriegeria” or “Kriegeria elatina’;

his only relevant publication was Winter (1881) where he synonymized

Ombrophila kriegeriana under Peziza elatina (as discussed above). Kriegeria

324 ... Toome, Pennycook, & Aime

Hohn. cannot be treated as a validation of “Kriegeria” Rabenh., because they

have different (albeit conspecific) generic types, Peziza elatina [= K. elatina]

versus “K. olivacea”. Consequently, Hohnel’s (1914) Kriegeria and K. elatina

must be interpreted as a gen. nov. and a comb. nov. attributable to Héhnel

alone (not to “Rabenh. ex Hohn. and definitely not to “G. Winter ex Hohn.”).

Kriegeria Hohn. 1914 is an illegitimate later homonym of the basidiomycetous

Kriegeria Bres. 1891 — a genus that Hohnel himself (1909: 1157-1159) had

apparently accepted as legitimate. Hohnel (1914) included a second species,

Kriegeria urceolus (Fuckel) Hohn. [= Rutstroemia urceolus (Fuckel) W.L. White

(White 1941; Whetzel 1945)].

In his monographic treatment of Rutstroemia, White (1941) included the

erroneous synonym “Kriegeria Wint. Hedw. 17: 32, 1878” (copied from Hoéhnel

1914) in his generic nomenclator. He accepted the full R. elatina nomenclator

published by Rehm (1893), including O. kriegeriana and its recombinations,

but he added the fictitious “Kriegeria elatina Wint. Hedwigia 17: 32, 1878” (also

copied from Hohnel 1914).

Seaver (1943) proposed to “revive” the Rabenhorst genus “Kriegeria’, typified

by Ombrophila kriegeriana, which he recombined as Kriegeria kriegeriana.

Because of its different type, Seaver’s genus must be interpreted as an additional

later homonym, Kriegeria Rabenh. ex Seaver, a heterotypic synonym of

Kriegeria Hohn. Seaver’s paper has several puzzling aspects. Inexplicably, he

cited two different “protologue” references: “Rab. Fungi Eur. 2315. 1878 [sic]”

(i.e., Rabenhorst 1876) for Ombrophila kriegeriana and “Hedwigia 17: 32.

1878” (i.e., Rabenhorst 1878a) for the genus “Kriegeria” and for the species

“Kriegeria olivacea”. Seaver’s failure to observe that all three names were actually

published simultaneously by Rabenhorst (1876) suggests that he had not seen

the O. kriegeriana exsiccata label. Significantly, Seaver (1943) did not mention

Hohnel’s treatment of Kriegeria nor Winter’s and Rehm’s synonymization of

O. kriegeriana with Rutstroemia elatina, and he explicitly rejected White's

(1941) placement of Kriegeria in Rutstroemia. Seaver (1943) synonymized his

genus Kriegeria with Chloroscypha Seaver, and cited the protologue description

of Chloroscypha (Seaver 1931) as the validating description of Kriegeria.

In addition to the type (Rabenhorst’s immature exsiccata material), Seaver

(1943) recombined five other species that had all previously been included in

Chloroscypha (Seaver 1931, 1938), and two further species were recombined in

Kriegeria by Seaver (1951). Apart from Kriegeria kriegeriana (on a pinaceous

host: Abies), all of Seaver’s Kriegeria species (on cupressaceous or taxodiaceous

hosts: Calocedrus, Chamaecyparis, Cryptomeria, Juniperus, Thuja, Sequoia)

are currently accepted in Chloroscypha (Petrini 1982). Seaver seems to have

been unaware of the incongruence between Kriegeria sensu Hohnel/White

[= Rutstroemia] and Kriegeria sensu Seaver [= Chloroscypha].

Kriegeria Bres. vs. Kriegeria Hohn. & Kriegeria Rabenh. ... 325

Kriegeria Bres. (Kriegeriaceae, Kriegeriales, Microbotryomycetes)

Kriegeria Bres., Rev. Mycol. (Toulouse). 13: 14. 1891.

= Xenogloea Syd. & P. Syd., Ann. Mycol. 17: 44. 1919, nom. illeg. (superfluous).

TyPE: Kriegeria eriophori Bres., Rev. Mycol. (Toulouse). 13: 14. 1891.

= Zymoxenogloea D.J. McLaughlin & Doublés, Mycologia 84: 671. 1992.

Type: Zymoxenogloea eriophori D.J. McLaughlin & Doublés, Mycologia 84: 671. 1992.

Contemporary material of the sole species was distributed as exsiccata

specimens (Pazschke 1895; Kohlmeyer 1962). Sydow & Sydow (1919) thought

that Kriegeria “Winter (cfr. Hedwigia 1878, p. 32)” was a valid name, making

the basidiomycetous Kriegeria Bres. an illegitimate later homonym, for which

they proposed Xenogloea as a replacement name. Their miscitation of Winter

as the author and Rabenhorst (1878a) as the protologue suggests that they

were relying on Hohnel’s (1914) treatment rather than carefully considering

the original Rabenhorst (1876) protologue. With “Kriegeria” Rabenh. being

an invalid provisional name, Kriegeria Bres. is legitimate and Xenogloea is

superfluous and illegitimate. Doublés & McLaughlin (1992) treated Kriegeria

Bres. as a legitimate generic name, and described a new genus Zymoxenogloea

for the yeast-like anamorphic state. Toome et al. (2013) proposed Kriegeriaceae

and Kriegeriales as a new family and order typified by Kriegeria Bres.

Discussion

The acomycetous genus Kriegeria has been published three times (once

invalidly and twice illegitimately):

(1) as “Kriegeria” Rabenh. 1876, an invalid provisional name that no

mycologist during the next 38 years ever treated as a formal name:

its invalid status has been argued by several eminent mycologists,

e.g., by Dennis (1954, 1956) and, most persuasively, by Donk (1958),

and it is not listed by Kirk et al. (2008) nor by the Index Nominum

Genericorum website (ING 2015);

(2) as Kriegeria Hohn. 1914, a valid generic name based on a different

type, but an illegitimate later homonym (of Kriegeria Bres.; ING

2015): it was not taken up by any subsequent mycologist but almost

universally accepted as a synonym of Rutstroemia (e.g., Rehm 1893,

White 1941, Whetzel 1945, Dennis 1956, Kirk et al. 2008) — major

errors in Hohnel’s protologue have created citational confusion that

still persists 100 years later;

(3) as Kriegeria Rabenh. ex Seaver 1943, an illegitimate later homonym

based on Rabenhorst’s invalid name: Seaver’s genus was rejected by

subsequent mycologists (e.g., Dennis 1954, 1956; Kobayashi 1965;

Petrini 1982), who argued that the species included in Kriegeria by

326 ... Toome, Pennycook, & Aime

Seaver (1943, 1951) were not congeneric with the type (Ombrophila

kriegeriana [= Rutstroemia elatina]) but instead belonged in

Chloroscypha. Seaver's Kriegeria homonym is not mentioned by Kirk

et al. (2008) nor by ING (2015).

There is no legitimately published ascomycetous genus Kriegeria, and this name

is not in current use.

In contrast, the basidiomycetous Kriegeria Bres. 1891 is a legitimate

monotypic genus that is predated only by the invalid “Kriegeria” Rabenh.

1876. Saccardo (1892: 497) tentatively identified the type as a Septogloeum,

and Hoéhnel (1909: 1157-1159) recombined it in Platygloea, but subsequently

Kriegeria Bres. has been accepted as a distinct genus, either under the

superfluous replacement name Xenogloea (Sydow & Sydow 1919: 44; Weiss

1950: 295; Martin 1952: 88-89; Kao 1956; Bandoni 1957: 838) or as a legitimate

Kriegeria Bres. (e.g., Davis 1922: 410; Donk 1958: 206; Oberwinkler & Bandoni

1982: 1744; Farr et al. 1989: 753; Doublés & McLaughlin 1992; Fell et al. 2001;

Swann et al. 2001; Sampaio & Oberwinkler 2011; Toome et al. 2013). Kriegeria

is accepted as legitimate, and Xenogloea as superfluous, by Kirk et al. (2008)

and ING (2015).

Acknowledgments

This study was supported by Assembling the Fungal Tree of Life (AFTOL) project

NSF DEB-0732968. The authors are grateful to Paul Kirk for numerous discussions and

to Drs. David McLaughlin and Dennis Desjardin for acting as expert referees.

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MY COTAXON

http://dx.doi.org/10.5248/130.329

Volume 130, pp. 329-336 April-June 2015

New records of crustose lichens and

a lichenicolous Arthonia from Vietnam

SANTOSH JOSHI’, DALIP KUMAR UPRETI’, SOON-OK OH’,

Tur THuy NGUYEN?, ANH DZUNG NGUYEN 3, & JAE-SEOUN HurR’*

' Lichenology Laboratory, CSIR-National Botanical Research Institute,

Rana Pratap Marg, Lucknow (UP)-226001, India

? Korean Lichen Research Institute, Sunchon National University, Suncheon-540 950, Korea

> Institute of Biotechnology & Environment, Tay Nguyen University,

567 Le Duan, Buon Ma Thuot City, Lak Province, Vietnam

*CORRESPONDENCE TO: jshur1@sunchon.ac.kr

ABSTRACT — New records from Vietnam of the crustose lichen species Arthonia excipienda,

Chiodecton leptosporum, Graphidastra multiformis, Pertusaria pycnothelia, P. thwaitesii, and

Phlyctis uncinata are presented together with the second world report of the non-lichenized

lichenicolous fungus, Arthonia diorygmae. All species were collected from the Bidoup Nui Ba

National Park located in the central highlands in Vietnam. A taxonomic description of each

species is accompanied by distributional and ecological data and illustrations.

KEY worps — corticolous, Da Lat city, evergreen forest, tropical

Introduction

A lichenological expedition was organized in 2014 by the National Research

Foundation of Korea and the Korea National Research Resource Center

Program. During fieldwork two of the authors collected a large number of

crustose lichens growing predominantly on trees in evergreen forests of

the Bidoup Nui Ba National Park in Vietnam. Among the more interesting

collections, we report and describe six corticolous species new for the country.

New Vietnamese records include Arthonia excipienda, first representatives

of the genera Chiodecton (C. leptosporum), Graphidastra (G. multiformis),

and Phlyctis (P. uncinata), and two new Pertusaria species (P pycnothelia,

P. thwaitesii) to be added the previously reported P. asiana Vain. and P. pertusa

(L.) Tuck. (Aptroot & Sparrius 2006, Joshi et al. 2014).

The lichenicolous non-lichenized Arthonia diorygmae was found

parasitizing a thallus of Diorygma, where it restricted the growth of the

330 ... Joshi & al.

host ascocarps. Although Arthonia accolens Stirt., A. antillarum (Fée) Nyl.,

A. cinnabarina (DC.) Wallr., A. cyanea Mill. Arg., and A. microcephala Vézda

have been previously recorded from Vietnam as corticolous and foliicolous

species (Aptroot & Sparrius 2006, Nguyen et al. 2011), no lichen-inhabiting

fungus has ever been reported from Vietnam in Arthonia or any other genus.

Materials & methods

Morphological features of specimens collected from tropical rain forests of Da Lat

city in Lam Dong province of Vietnam were examined under Magnus Zoom Stereo

Trinocular (MSZ-TR) dissecting microscope and anatomical characters were observed

under a compound microscope (Leica DM 500). Chemical spot tests and TLC (using

solvent system A) were conducted according to Orange et al. (2010). The studied

material is deposited in the herbarium of KoLRI (Korean Lichen Research Institute),

South Korea.

Taxonomy

Arthonia diorygmae S. Joshi & Upreti, Lichenologist 45: 323, 2013. PL.1A

Thallus absent, non-lichenized, lichenicolous on Diorygma; ascomata dot-

like, round, irregular to shortly lirellate, prominent; 0.2-0.3 x 0.1-0.2 mm;

proper exciple completely carbonized to dark brown, 15-20 um wide, in

continuation with hypothecium, undulating, V-shaped; epihymenium brown,

15-25 um high; hymenium hyaline, inspersed with oil droplets, gelatinous,

<100 um high, I+ wine red; paraphyses indistinct, 1-1.5 um thick; asci broadly

clavate, 8-spored, 40-50 x 13-15 um, epiplasm I+ wine red; ascospores hyaline

becoming dark brown and warty, 2-celled, ovoid to slipper-shaped, 10-16 x

4—5 um, I-.

CHEMISTRY: Ascomata (Arthonia): K-, PD-, C-. Thallus (Diorygma

host species): K+ yellow, PD+ yellow-orange, C-; norstictic and stictic acid

chemosyndrome detected in TLC of host species.

DISTRIBUTION & ECOLOGY: This is the second report of A. dorygma,

previously known only from the type locality in India (Joshi et al. 2013b).

In Vietnam it was collected at ca. 1400 m from a thick- and smooth-barked

tree, where it was growing luxuriantly and spread largely in association with

Cryptothecia, Diorygma, and other graphidaceous taxa.

SPECIMENS EXAMINED: VIETNAM. LAM DONG PROVINCE: Bidoup Nui Ba National

Park, Da Lat city, 12°10°55.4”N 108°40’50.1”E, alt. 1454 m, on tree bark, 7 January 2014,

Hur & Oh VN140150, VN140098 (KoLRI).

REMARKS: This species is distinctive in having non-lichenized lichenicolous life

form. Though, our specimen differs from the A. diorygmae type material in

also producing stictic acid by the host, all taxonomic characters were similar

to those of the holotype described from the Western Ghats in India (Joshi et

al. 2013b).

New crustose lichen records (Vietnam) ... 331

Arthonia excipienda (Nyl.) Nyl., Lich. Scand.: 261, 1861. PL. 1B

Thallus corticolous, crustose, endoperidermal, smooth, continuous, shiny,

pale green to yellowish, thin, 40-60 tm in cross section, corticate; cortex

indistinct to 30 um; photobiont cells chlorococcoid, aggregate to disperse,

layer discontinuous, <25 um; medulla indistinct; ascomata lirelliform, sessile;

lirellae 0.5-1 x 0.1-0.2 mm; disc slightly open, black, epruinose; thalline

margin absent to indistinct; proper exciple laterally carbonized, 25-30 um

wide, convergent, internally lined by 20-60 um periphysoids; epihymenium

brown, 10-15 um high; hymenium hyaline, clear, gelatinous, 60-70 um high,

I+ blue; hypothecium hyaline to brownish 20-25 um high; asci broadly clavate,

8-spored, 60-65 x 20-22 um; ascospores hyaline to brown, 19-21 x 7-8 um, I-.

CHEMISTRY: K-, PD-, C-; no lichen compounds detected in TLC.

DISTRIBUTION & ECOLOGY: The species was recorded from West European

countries and South Korea (Joshi et al. 2013a); in Vietnam, it was found

growing poorly at 1400-1500 m in association with other crustose corticolous

lichens on smooth-barked trees.

SPECIMENS EXAMINED: VIETNAM. Lam DONG PROVINCE: Bidoup Nui Ba National

Park, Da Lat city, 12°10’55.4”N 108°40’50.1”E, alt. 1454 m, on tree trunk, 7 January

2014, Hur & Oh VN140080 (KoLRI); 12°10’54.6”N 108°40’45.1’E, alt. 1426 m, on tree

trunk, 8 January 2014, Hur & Oh VN140167 (KoLRI).

REMARKS: Arthonia excipienda, an uncommon species in Asia, apparently

resembles A. punctiformis Ach. and A. radiata (Pers.) Ach., which differ in

having multiseptate ascospores. Arthonia didyma Korb. also produces 2-celled

ascospores, but they are brown and warty on maturity.

Chiodecton leptosporum Miill. Arg., Flora 65: 332, 1882. PL. 1c

Thallus corticolous, crustose, epiperidermal, tightly attached to the substrate,

smooth to slightly verruculose, pale green, with a white pruina, 200-400 um

thick in cross section, ecorticate; photobiont cells Trentepohlia-like, layer <200

um; medulla whitish with few calcium oxalate crystals, 200-250 um, mostly

endoperidermal; prothallus usually distinct, brownish; ascomata perithecioid,

dot-like to round, solitary or rarely united, aggregated into distinctly elevated

stroma-like structures usually with more than 10 ascocarps, 0.1-0.2 mm in

diameter; stroma round to oval, 0.5-2 mm in diameter; proper exciple dark

brown to completely carbonized, 25-45 um; epihymenium brown, indistinct to

15 um high; hymenium hyaline, clear, 135-140 um high, I+ blue; hypothecium

extending down to the substrate, hard and black, <200 um high; paraphyses

1-1.5 um thick; asci clavate, 8-spored, 80-100 x 9-12 um, I-; ascospores

obovate, hyaline, 3-septate, 30-40 x 3-4 um, I-; locules cylindrical, 7-9 x 1-2

um.

CHEMISTRY: K-, PD-, C-; roccellic acid detected in TLC.

332 ... Joshi & al.

DISTRIBUTION & ECOLOGY: Chiodecton leptosporum is widely distributed

in Asia, Australia, and scattered localities in New Caledonia, Fiji, and Guam

(Thor 1990); in Vietnam, it was luxuriantly growing at 1400-1500 m on hard-

barked trees in an evergreen forest. The other lichens growing in association

were unknown leprose taxa and Cryptothecia.

SPECIMENS EXAMINED: VIETNAM. Lam DONG PROVINCE: Bidoup Nui Ba National

Park, Da Lat city, 12°10’55.4”N 108°40’50.1”E, alt. 1454 m, on tree bark, 7 January

2014, Hur & Oh VN140043, VN140065, VN140067, VN140078, VN140090 (KoLRI);

12°10’38.9"N 108°40’37.9’E, alt. 1426 m, on tree bark, 8 January 2014, Hur & Oh

VN140149 (KoLRI).

REMARKS: Chiodecton leptosporum can easily be confused with another most

commonly distributed C. congestulum Nyl., which differs in a thallus containing

a yellow pigmented medulla and a hymenium with a feeble iodine reaction.

A comparatively rougher thallus and mostly united ascocarps further

distinguish C. congestulum from C. leptosporum.

Graphidastra multiformis (Mont. & Bosch) G. Thor, Opera Bot. 103: 82, 1991.

Pu, Lp

Thallus corticolous, crustose, epiperidermal, tightly attached to the

substrate, cretaceous, pale green to greenish grey, 200-300 um thick in cross

section, epinecral layer resembling cortex is present, 80-85 um; photobiont

cells Trentepohlia-like, layer <120 um; medulla whitish, with few to numerous

calcium oxalate crystals, 100-120 um; prothallus whitish in inner part and

brownish in outer part; ascomata usually lirelliform, but sometimes apothecioid,

solitary; apothecioid and lirelliform structures distinctly elevated with a slightly

to distinctly constricted base, 1-2 x 0.5-1 mm; disc black, 5-10 mm thick;

thalline margin 130-330 um thick; proper exciple thinly carbonized laterally,

15-20 um, basally continuous with hypothecium; epihymenium brown, 25-35

um; hymenium hyaline, clear, 60-85 um, I+ reddish; hypothecium extending

down to the substrate, hard, black, + V-shaped, 0.3-0.5 mm; paraphyses 1-2

um thick; asci clavate, 55-60 x 5-6 um, I+ blue; ascospores hyaline, spermatoid,

3-septate, 35-40 x 2-3 um (including tail of 20-22 um), I-.

CHEMISTRY: K-, PD-(?), C-; roccellic acid and traces of protocetraric acid

detected in TLC.

DISTRIBUTION & ECOLOGY: Graphidastra multiformis has previously been

reported from India, Sri Lanka, The Philippines, Australia, West Samoa, and

Tahiti (Thor 1990); in Vietnam it was collected at ca. 1400 m from thick- and

hard-barked trees, where it was growing luxuriantly along with members of

Pannariaceae and Chiodecton leptosporum.

SPECIMEN EXAMINED: VIETNAM. LAM DONG PROVINCE: Bidoup Nui Ba National

Park, Da Lat city, 12°10°55.4”N 108°40’50.1”E, alt. 1454 m, on tree bark, 7 January 2014,

Hur & Oh VN140075 (KoLRI).

New crustose lichen records (Vietnam) ... 333

REMARKS: Graphidastra multiformis is close to G. byssiseda (Mill. Arg.) G.

Thor in having spermatoid ascospores and a thallus containing roccellic acid,

but G. byssiseda differs in producing bi-clavate larger ascospores and lacking

protocetraric acid.

Pertusaria pycnothelia Nyl., Bull. Soc. Linn. Norm., ser. 2, 2: 70, 1868. PL. 1E

Thallus corticolous, crustose, greenish grey to green or pale green, shiny,

+ rimose, verrucose, <500 um thick in cross section, corticate; cortex 20-25

um; photobiont Trebouxia, layer 85-90 um; medulla 180-320 um; prothallus

blackish; ascomata verruciform, numerous, scattered, sometimes confluent and

forming an almost continuous crust, concolorous with the thallus, flattened,

hemispherical, 0.5-0.8 mm in diam; ostiole conspicuous, brown, dark brown

in apothecioid to blackish in verruciform, 0.3-0.5 mm in diam; thalline

margin 300-370 um wide; proper exciple hyaline to yellowish, 25-50 um wide;

epihymenium pale brown, 45-47 um high; hymenium hyaline, clear, 300-400

um high; subhymenium hyaline, 80-120 um high; paraphyses 1-2 um thick;

asci broadly clavate, 2-spored; ascospores hyaline, ellipsoid to subfusiform,

double walled, wall smooth, 50-75 x 17-25 um, I+ blue.

CHEMISTRY: K-, KC-, C-, PD-; 4,5-dichlorolichexanthone and

2’-O-methylperlatolic acid detected in TLC.

DISTRIBUTION & ECOLOGY: Pertusaria pycnothelia was previously reported

from Australia, New Caledonia, and Papua New Guinea (Archer 1997); in

Vietnam, it was collected at 1400-1500 m, where it was growing in small

patches over tree bark in an evergreen forest.

SPECIMEN EXAMINED: VIETNAM. LAM DONG PROVINCE: Bidoup Nui Ba National

Park, Da Lat city, 12°10°55.4”N 108°40’50.1”E, alt. 1454 m, on tree bark, 7 January 2014,

Hur & Oh VN140096 (KoLRI).

REMARKS: The chemically similar Pertusaria trachyspora A.W. Archer is

distinguished from P. pycnothelia by ascospores with a rough inner wall.

Pertusaria thwaitesii Mill. Arg., Flora 67: 470, 1884. PL. 1F

Thallus corticolous, crustose, epiperidermal, wrinkled, cracked, uneven,

shiny, off-white to pale grey, 100-150 um thick in cross section, corticate; cortex

10-15 um; photobiont Trebouxia, layer 50-80 um; medulla white, 70-90 um;

prothallus jet black; ascomata verruciform, conspicuous, concolorous with the

thallus, numerous, very flattened hemispherical or irregular in outline, 1-2 mm

wide; ostioles numerous, conspicuous, pale yellow becoming black, punctiform,

becoming somewhat sunken, 0.05-0.07 mm in diam., 4-7 per verruca; proper

exciple hyaline to yellowish, indistinct to 30 um wide; epihymenium indistinct;

hymenium hyaline, clear, <430 um high; subhymenium indistinct; asci broadly

334 ... Joshi & al.

clavate, 2-spored, 290-300 x 45-50 um; ascospores elongate ellipsoid, double

walled, inner wall ornamented and outer wall smooth, 140-165 x 35-45 um.

CHEMISTRY: K-, PD+ orange-red, C-; protocetraric acid detected in TLC.

DISTRIBUTION & ECOLOGY: Known from Australia, Papua New Guinea,

Sri Lanka (Archer 1997); in Vietnam, Pertusaria thwaitesii was growing in

association with Graphis at 1700-1800 m in trees in an evergreen forest.

SPECIMEN EXAMINED: VIETNAM. LAM DONG PROVINCE: Mt Langbian, Da Lat city,

12°02’18.6’N 108°25’35.1”E, alt. 1779 m, on tree bark, 9 January 2014, Hur & Oh

VN140281 (KoLRI).

REMARKS: Pertusaria hartmannii Mull. Arg., which superficially resembles

P. thwaitesii, differs in producing smoother ascospores and a thallus containing

norstictic acid (Archer 1997).

Phlyctis uncinata Stirt., J. Linn. Soc., Bot. 14: 464, 1875. PL. 1G

Thallus corticolous, crustose, rough, epiperidermal, subleprose, + rimose

due to substrate, greenish grey, whitish green to greyish green, 100-130 um

thick in cross section, ecorticate; photobiont cells green protococcoid, layer

50-70 um; medulla white, 40-50 um; prothallus white; ascomata round to

irregular, solitary to aggregate, immersed, 0.4-0.5 mm; disc blackish, finely

pruinose, 0.2-0.3 mm in diameter; thalline margin recurved, 165-180 um wide;

proper exciple hyaline to brownish, apically dark, 35-55 um wide; epihymenium

granular, brownish 12-15 um high; hymenium hyaline, clear, 65-80 um high,

I+ wine red; subhymenium <40 um high; asci clavate, 8-spored, 110-165 x

15-25 um, I+ wine red; ascospores hyaline, fusiform, crescent shape,

transversely septate, 50-54 x 5-7 um, 8-locular, I+ wine red.

CHEMISTRY: K+ yellow turning red, PD+ yellow-orange, C-; norstictic acid

detected in TLC.

DISTRIBUTION & ECOLOGY: Known from New Zealand and Thailand

(Galloway 1985, 2007); in Vietnam, the species was collected at 1700-1800 m,

where it was spread in short patches on rough-barked trees.

SPECIMEN EXAMINED: VIETNAM. LAM DONG PROVINCE: Mt Langbian, Da Lat city,

12°02’18.6”N 108°25’35.1”E, alt. 1779 m, on tree bark, 9 January 2014, Hur & Oh

VN140109 (KoLRI).

REMARKS: Phlyctis uncinata is closely related to P. karnatakana S. Joshi & Upreti,

which differs in having small (20-30 um long) ascospores (Joshi et al. 2010).

The thallus of Phlyctis himalayensis (Nyl.) D.D. Awasthi is morphologically

similar but reacts K+ red (Awasthi 1991). Phlyctis, which has poorly delimited

species, needs further revision to clarify the status of P uncinata (Joshi & Upreti

2012; Joshi et al. 2010, 2012).

New crustose lichen records (Vietnam) ... 335

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Prate. 1. New records of lichens from Vietnam. A. Arthonia diorygmae (white arrow indicating

ascomata of Arthonia and black arrow indicating suppressed growth of ascomata in host);

B. Arthonia excipienda (ascomata); C. Chiodecton leptosporum; D. Graphidastra multiformis;

E. Pertusaria pycnothelia; F. Pertusaria thwaitesii; G. Phlyctis uncinata. Scale bars: A, F, G = 0.5 mm;

B, E=1mm;C,D=2 mm.

Acknowledgments

This work was supported by a grant from the National Research Foundation of Korea

(#2011-0031494) and the Korea National Research Resource Center Program. Authors

are grateful to Drs. S.Y. Kondratyuk and L. Lék6s for their valuable comments on the

manuscript. DKU and SJ thank Director, CSIR-National Botanical Research Institute,

Lucknow, India, for providing infrastructure to facilitate the identification of Korean

lichen material.

336 ... Joshi & al.

Literature cited

Aptroot A, Sparrius LB. 2006. Additions to the lichen flora of Vietnam, with an annotated checklist

and bibliography. Bryologist 109(3): 358-371.

http://dx.doi.org/10.1639/0007-2745(2006) 109[358:ATTLFO]2.0.CO;2

Archer AW. 1997. The lichen genus Pertusaria in Australia. Bibliotheca Lichenologica 69. 249 p.

Awasthi DD. 1991. A key to the microlichens of India, Nepal and Sri Lanka. Bibliotheca

Lichenologica 40. 337 p.

Galloway DJ. 1985. Phlyctella Krempelh. 385-389, in: Flora of New Zealand Lichens. Government

Printer, Wellington, New Zealand.

Galloway DJ. 2007. Phlyctis (Wallr.) Flot. 1184-1191, in: Flora of New Zealand Lichens, 2nd ed.,

Vol. 2. Manaaki Whenua Press, Lincoln, New Zealand.

Joshi S, Upreti DK. 2012. Lichen genus Phlyctis (Phlyctidaceae) in India. Geophytology 44(3):

363-369.

Joshi S$, Upreti DK, Mishra GK, Divakar PK. 2010. Two new species of lichen genus Phlyctis in

India. Bryologist 113(4): 724-727. http://dx.doi.org/10.1639/0007-2745-113.4.724

Joshi S, Upreti DK, Nayaka S. 2012. Two new species in the lichen genus Phlyctis (Phlyctidaceae)

from India. Lichenologist 44(3): 363-369. http://dx.doi-org/10.1017/S0024282911000879

Joshi S, Kondratyuk SY, Crisan F, Jayalal U, Oh SO, Hur JS. 2013a. New additions to lichen mycota

of the Republic of Korea. Mycobiology 41(4): 177-182.

http://dx.doi.org/10.5941/MYCO.2013.41.4.177

Joshi S, Upreti DK, Nayaka S. 2013b. A new lichenicolous Arthonia species (Arthoniaceae) on

Diorygma from India. Lichenologist 45(3): 323-327.

http://dx.doi.org/10.1017/S0024282913000042

Joshi S, Jayalal U, Oh SO, Nguyen TT, Dzung NA, Hur JS. 2014. A new species of Graphis and new

lichen records from Vietnam, including a second worldwide report of Sarcographina cyclospora.

Mycobiology 42(1): 17-21. http://dx.doi.org/10.5941/MYCO.2014.42.1.17

Nguyen TTT, Joshi Y, Licking R, Nguyen AD, Wang XY, Koh YJ, Hur JS. 2011. Seven new records

of foliicolous lichens from Vietnam. Mycotaxon 117: 93-99. http://dx.doi.org/10.5248/117.93

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

British Lichen Society, London.

Thor G. 1990. The lichen genus Chiodecton and five allied genera. Opera Botanica 103. 92 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.337

Volume 130, pp. 337-359 April-June 2015

First records of some Asian macromycetes in Africa

PaBLo P. DANIELS’, OUMAROU HAMA?’, ALFREDO JUSTO FERNANDEZ?,

FELIX INFANTE GARCIA-PANTALEON’, MoussA BARAGE?’,

DAHIRATOU IBRAHIM‘, & MARIA ROSAS ALCANTARA!

"Department of Botany, Ecology and Plant Physiology, University of Cordoba,

Ed. Celestino Mutis, Campus Rabanales, Cordoba 14071 Spain

? Department of Plant Production, Faculty of Agronomy, University Abdou Moumouni,

Niamey BP-10960 Niger

* Biology Department, Lasry Biological Science Center, Clark University,

950 Main St., Worcester, MA 01610 USA

* Life Sciences and Earth Department, High School of Education, University Abdou Moumouni,

Niamey BP-10963 Niger

* CORRESPONDENCE TO: ppdaniels@hotmail.com

ABSTRACT — This paper reports and discusses preliminary data on new Asian macromycete

species now recorded on the African continent and collected for the first time in Niger

during sampling conducted in the southwestern region from 2008 to 2012. Descriptions and

comments on chorology, systematics, and closely related species are given for Hymenagaricus

subepipastus, Clitopilus orientalis, Tulostoma evanescens, Termitomyces bulborhizus, and

Volvariella cf. sathei.

Key worps — basidiomycetes, fungi, taxonomy

Introduction

The literature available on the macromycetes of West Africa is generally both

sparse and highly fragmented. Boa (2004) reported that there appeared to be no

data at all for Niger, but since then a handful of references in local publications

and international journals have cited a total of 16 fully identified species for

this country (TABLE 1). Countries bordering this sub-Saharan region (with

the exception of Mali and Chad) have recently been studied by mycologists

who are beginning to publish research on the diversity, systematics, ecology,

ethnomycology and use of macromycetes. As a result, much more information

is now available on fungal species in Benin (De Kesel et al. 2000, 2002; De Kesel

& Yorou, 2000; Yorou et al. 2002a,b; Yorou & De Kesel 2002; Yorou, 2010),

Burkina Faso (Ganaba et al. 2002, Guissou et al. 2002, 2008, Guissou 2005),

338 ... Daniéls & al.

TABLE 1. Macromycete species previously recorded from Niger.

TAXON REFERENCE

Agaricus augustus Fr. Hama et al. 2008

Agaricus bulbillosus Heinem. & Gooss.-Font. Hama et al. 2008

Agaricus subsaharianus L.A. Parra et al. Hama et al. 2010, 2012

Crinipellis glaucospora (Beeli) Pegler Antonin 2013a

Crinipellis pseudosplachnoides (Henn.) Singer Antonin 2013a

Ganoderma colossus (Fr.) C.F. Baker Hama et al. 2008, 2009, 2012

Ganoderma lucidum (Curtis) P. Karst. Hama et al. 2008, 2009

Itajahya rosea (Delile) E. Fisch. Hama et al. 2008

Leucocoprinus brebissonii (Godey) Locq. Hama et al. 2008

Lyophyllum aggregatum (Schaeff.) Kihner Hama et al. 2008

Marasmius atrorubens (Berk.) Mont. Antonin 2013b

Phellinus allardii (Bres.) S. Ahmad Hama et al. 2012

Podaxis pistillaris (L.) Fr. Hama et al. 2008, 2009, 2012

Termitomyces striatus (Beeli) R. Heim Hama et al. 2008, 2009

Trametes cingulata Berk. Hama et al. 2008

Trametes leonina (Klotzsch) Imazeki Hama et al. 2008

Nigeria (Zoberi 1973; Oso 1975, 1977; Adewusi et al. 1993; Osagualekhor &

Okhuoya 2005; Akpaja et al. 2003; Osemwegie et al. 2006; Okhuoya et al. 2010)

and, to a lesser extent, Togo (De Kesel et al. 2008, Gardt et al. 2011) and Ivory

Coast (Koné et al. 2010a,b). In general, many West African ecosystems are very

sparsely explored. However, Yorou et al (2014) recently updated a total list of

about 72 edible macromycetes for West Africa (Benin, Burkina Faso, Ivory

Coast, and Togo). In West Africa, macromycetes are not only taxonomically

poorly documented, but patterns of distributions, ecology, and local use should

be addressed especially in the context where forest and savannah ecosystems

are disappearing at an alarming rate (FAO 2010) along with the fungal species.

As an example, a preliminary Red List of threatened fungi has been published

for Benin (Yorou & De Kesel 2011).

Given the lack of information regarding Niger, the University of

Cordoba submitted to the Spanish Agency for International Development

Cooperation a project to be implemented jointly with the Abdou Moumouni

University, Niamey, entitled “Edible and cultivable Macromycetes of Niger

(Ethnomycology)”. One of the aims of this project was to draw up a classified

inventory of Niger's fungal biodiversity. At an earlier stage, the local project

team had already compiled a preliminary inventory, which was expanded

and completed as part of the current research and served as a basis for the

subsequent ethnomycological survey. A number of collections have been

published recently (Hama et al. 2012, Antonin 2013a,b). The findings of the

present study relate to the fungal diversity portion of the project and focus on

the key taxonomic features of species reported earlier in Asia but which are

considered first records in Africa.

New Asian macromycete records for Africa ... 339

Materials & methods

Material collected between 2008 and 2012 came mainly from the West Sudanian

savannah (White 1986), deemed the most suitable area for a study of national

macromycete biodiversity in view of its abundant vegetation; the study centered

primarily on the deep-soil, wooded savannah and gallery forests bordering rivers of

the W National Park in southwestern Niger. Mushrooms were mostly collected using

routine sampling methods (Halling 1996, Eyi-Ndong et al. 2011); each species was

assigned a collection number and photographed with a Canon 400D, Canon PowerShot

G10, or Olympus U 700 digital camera. Fresh mushroom organoleptic data - colour,

odour, and taste - and information on gross features likely to be modified during

drying and required for identification purposes were entered on field records. Collected

mushrooms were then dehydrated in situ using a Bunsen burner attached to a folding

portable dryer (De Kesel 2001) and then put inside minigrip bags. Exsiccata were kept

at the Abdou Moumouni University (Niamey, Niger) and duplicates were placed in the

fungal section of the COFC herbarium (Thiers 2014). Some exsiccata and type material

from K and BR herbaria were requested as loans to compare with Nigerien samples and

also with protologues.

Colour references were coded following Kornerup & Wanscher (1981). Material was

then studied under a Nikon Labophot2 light microscope fitted with a drawing tube at

a 1000x magnification. Spore measurements were made in 3% KOH mounts (Menzel-

Glaser) in profile position and excluding the hilar appendix and ornamentation (Lm

= mean length; Wm = mean width, Em = Lm/Wm). Unless otherwise indicated, the

specimens were identified by PP. Daniéls. Species citations follow Index Fungorum

(www.indexfungorum.org). Collection localities are placed in alphabetic order. For

each collection, field data are ordered as follows: Country, Administrative Region,

Department, Locality, Site, {Park}, latitude/longitude coordinates, altitude, ecology

and substrate, date, collector and identifier (where appropriate), collection accession

number, and herbarium accession number. Distribution notes have been added when

this was significant compared to other African flora, and observations are included on

nomenclature and taxonomy when these differ from known species descriptions.

Standard methods for DNA extraction, PCR amplification, and DNA sequencing

were applied (e.g., Justo et al. 2011) to check Nigerien Volvariella samples due to the

noteworthy morphological differences found with closely related species. Primer pairs

ITS1F and ITS4 (Gardes & Bruns 1993) were used for both PCR and sequencing. The

remaining sequences used in the analysis were retrieved from GenBank and come from

the studies of Menolli & Capelari (2008), Li et al. (2009), and Justo et al. (2011). GenBank

accession numbers are given under the specimens examined section. Volvariella

bombycina (Schaeff.) Singer and V. volvacea (Bull.) Singer were used as outgroup taxa in

the final dataset. These two species appear to be the sister group to all other Volvariella

species sequenced to date (Vizzini et al. 2011). Sequences were aligned with MAFFT

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

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

deposited in TreeBASE (http://purl.org/phylo/treebase/phylows/study/TB2:S15058). A

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

100 rapid bootstrap replicates.

340 ... Daniéls & al.

Taxonomy

Clitopilus orientalis TJ. Baroni & Watling, Mycotaxon 72: 58, 1999. PLATE 1

MACROCHARACTERS — BASIDIOMATA small, gregarious. PILEUS 1-2.2 cm

diameter, at first convex, then flat to centrally depressed; white coloured [A1]

and fibrillose; margin involute and radially ridged. St1pe central or slightly

eccentric, slightly turbinate, up to 10-19 x 2-4 mm, concolourous with pileus,

surface pruinose, base with white mycelial tufts. LAMELLAE subdecurrent to

decurrent, with lamellulae, 3-4/mm, white at first [A1], then brownish pink

[5A3, 5B5], with whitish crenulate margin. CONTEXT fibrous; odour and taste

not recorded.

1000000000008 |:

cri

SUNY

Pate 1. Clitopilus orientalis (COFC-F 5160):

a) Pileipellis; b) Basal mycelium; c) Pileus context;

d) Spores; e) Basidia; f) Cheilocystidia; g) Stipitipellis showing a secretory hyphae.

New Asian macromycete records for Africa ... 341

MICROCHARACTERS — PILEIPELLIS made up of interlaced 3-5 um wide

elements forming a trichodermis; hyphae hyaline, thin-walled. STiprT1PELLIs

similar to pileipellis but with inner hyphae parallel, 3-13 um wide; hyphae

hyaline, thin-walled, with conspicuous septa. CONTEXT HYPHAE subparallel,

cylindrical, 3-8 um wide, thin-walled, sometimes with vesicular end, 12-14 um

wide, hyaline and thick-walled. MycELi1aL TuFTs with interlaced cylindrical

hyphae, 4-6.5 um wide, hyaline, thin-walled. Secretory hyphae filled with

refringent content, 5-13.5 um wide. CLAMP CONNECTIONS absent in all

structures. CHEILOCYSTIDIA cylindrical to fusiform or subcapitate, 29-37(-42)

x 4-8 um. PLEUROCYsTIDIA absent. Basrp1a claviform, 21-30 x 7-9 um, with

4 sterigmata, thin-walled. Sporss ellipsoid, striate with 10-11 longitudinal

ribs, hyaline, thin to thick-walled, (6.3-)6.8-7.5(-8.5) x (3.7-)4-4.7(-5) um,

[Lm = 7.1 um, Wm = 4.4 um, E = 1.40-1.88; Em = 1.63].

SPECIMEN EXAMINED — NIGER, Dosso, Gaya, Albarkeizé, Saboula, 12°04’55.6”N

3°14’07.5’E, elevation 136 m, fallow land beside the Niger river on a termite mound

under Anacardium occidentale L., 13 September 2008, O. Hama, Hama 132 (COFC-F

5160).

EcoLoGcy & DISTRIBUTION — On termite mounds in India and Malaysia

(Baroni & Watling 1999). Clitopilus orientalis appears not to have been

previously reported in Africa.

ComMEnts — ‘This particular Clitopilus species displays cheilocystidia and

grows on termite mounds (Baroni & Watling 1999). The specimen examined

differs from descriptions of the Asian species only in that cheilocystidia are

abundant and sometimes clustered, rather than scattered. The most closely

related species are C. apalus (Berk. & Broome) Petch, which has subglobose

rather than ellipsoid spores, and C. peri (Berk. & Broome) Petch, which neither

grows on termite mounds nor displays cheilocystidia (Pegler 1977, Baroni &

Watling 1999).

Hymenagaricus subepipastus Heinem. & Little Flower, Bull. Jard. Bot. Nat. Belg.

54: 168, 1984. PLATES 2, 3

MACROCHARACTERS — BASIDIOMATA small, gregarious. PILEUS 0.5-1.2

cm diameter, at first convex, then trapezoid to flattened displaying dark

olive-green [1E7, 30D7] scales on a light-green background [30A4], changes

colour slightly during ageing, the background turns yellow [4A4] while the

scales — though retaining their colour — become less abundant, since they

are easily detached; margin thin, regular. Stipe central, fistulose, cylindrical,

concolourous with pileus, whitish towards the base, up to 22 x 0.6 mm, surface

pruinose to squamulose with scarce evanescent scales below ring zone, base

342 ... Daniéls & al.

PLATE 2. Hymenagaricus subepipastus (COFC-F 5186): Fresh basidiomata. Scale bar = 1 cm.

with white mycelial tufts. RING absent or powdery and evanescent, seen only in

very young specimens. LAMELLAE free, with lamellulae, 3-4/mm, greyish white

at first [B2], then grayish yellow [1B2-3, 2B2-3], and finally brown [5C7], with

paler crenulate margin. Spore deposit brown [5C7]. ConTExtT thin, fibrose, in

stipe; odour and taste not recorded.

MICROCHARACTERS — PILEIPELLIS made up of claviform, pyriform to

spherical elements 7-17 x 7-16 um grouped in chains and tufts forming a

hymeniodermis, with thick walls and encrusted brownish orange pigments;

subpellis made up of cylindrical elements, interwoven to subparallel,

3.5-8 um, also with a thick wall and encrusted brownish orange pigments,

becoming thinner, parallel and hyaline inwards. StT1p1TIpELLis formed by

parallel cylindrical elements, 3-9 um wide, often with chains of doliiform

(barrel-shaped) to short-claviform terminal elements grouped in tufts; hyphae

hyaline to yellowish brown, thin- to thick-walled, somewhat constricted at the

septa. CONTEXT HYPHAE subparallel, cylindrical, 4-12 tm wide, thin-walled.

MYCELIAL TUFTS with parallel cylindrical hyphae of 2-3.5(-5) um wide, hyaline

and thin-walled, sometimes with secretory hyphae, filled with refringent

contents. CLAMP CONNECTIONS absent in all structures. CHEILOCYSTIDIA

lageniform to fusiform, 16-20(-26) x (5-)6-10(-12) um. PLEUROCYSTIDIA

absent. Basip1a widely claviform, 12-15 x 5-7 um, with (2)4 sterigmata,

thin-walled. Spores ovoid to ellipsoid, smooth, brown coloured, thin to

New Asian macromycete records for Africa ... 343

d DodagaaDoaaao lz

6 IDUUDO § cess

PRU

PLATE 3. Hymenagaricus subepipastus (COFC-F 5186):

a) Pileipellis; b) Basal mycelium; c) Pileus context; d) Spores;

e) Basidia; f) Cheilocystidia; g) Stipitipellis.

thick-walled, (4-)4.5-6.3 x (2.7-)3-4(-4.5) um, [Lm = 5.1 um, Wm = 3.4 um,

E = 1.29-1.71; Em = 1.49].

SPECIMEN EXAMINED — NIGER, Niamey, Niamey, Karadjé, guest house at Abdou

Moumouni University, 13°29’25.5”N 2°04’64’E, elevation 200 m, in garden soil, 8

August 2010, P. Daniéls, Hama 383 (COFC-F 5186).

344 ... Daniéls & al.

EcoLocy & DISTRIBUTION — Scattered in the grass or garden soils.

Hymenagaricus subepipastus was first described in India (Heinemann & Little

Flower 1984), but Heinemann (1986) considered its presence in Africa likely.

Our report may be the first record of the species in Africa.

COMMENTS — Our specimen fully matches existing descriptions of

H. subepipastus by Heinemann & Little Flower (1984). This species is related to

H. viridulus Heinem. & Little Flower, but the spores of the studied collection

are larger and ovoid rather than cymbiform (wineskin-shaped). Hymenagaricus

subaeruginosus (Berk. & Broome) Heinem. & Little Flower has slightly smaller,

cymbiform spores and larger basidiomata, with a pileus up to 4.5 cm diameter

(Heinemann & Little Flower 1984). Hymenagaricus epipastus (Berk. & Broome)

Heinem. & Little Flower differs from H. subepipastus only in spore size, and has

been reported in Ceylon (Asia) (Heinemann & Little Flower 1984). For details

see TABLE 2.

TABLE 2. Comparison of pileus and spores from species related to

Hymenagaricus subepipastus.

SPECIES PILEUS DIAM. (cm) Lm Wm _ SPORE SHAPE

H. viridulus type* 0.5-1.5 4.7 3.3 Cymbiform

H. subaeruginosus isotype (K) 1.5-2.5°/ 4.5 47 3.2 Cymbiform

H. subaeruginosus MGF 5088 (BR) 1.5-2.5> 4.5 3.0 Cymbiform

H. epipastus type * 2 42—6 301 Ovoid

H. subepipastus type * 1-1.5 5.0 3.6 Ovoid

H. subepipastus Hama 383 0.5-1.2 5.l- 34 Ovoid

* Heinemann & Little Flower 1984; measurements in exsiccatum

Termitomyces bulborhizus T.Z. Wei, Y.J. Yao, B. Wang & Pegler, Mycol. Res. 108:

1458, 2004. PLATES 4-6

MACROCHARACTERS — BASIDIOMATA medium sized, gregarious. PILEUS

4-12 cm diameter, fleshy, at first convex, then flattened and slightly depressed

with obtuse blunt perforatorium 2-3 mm wide; white, isabella to olivaceous

brown coloured [A1, 4A4, 4B6, 5A3] pallescent towards the margin and with

brownish orange centre [6D5, 5B6-8]; surface smooth and dry or slightly viscose

and radially rugose in wet conditions; margin thin, sinuous to sub-lobate,

slightly involute when young then uplifted when old. St1px central, cylindrical,

straight, white to slightly isabella coloured [A1], (4—)7-11 x 0.5-2 cm, surface

squamulose with evanescent recurved 2-3 mm long scales; bulbose at ground

level of 2-3.5 cm diameter. PSEUDORHIzOID robust, rugose and grooved

longitudinally, orange brown [4B5, 5C5, 6C8, 6B7] to black [5E3, 6F2-5, F1]

downwards, of 5-20 cm long, tapering quickly towards the base. LAMELLAE

New Asian macromycete records for Africa ... 345

Piate 4. Termitomyces bulborhizus: fresh basidiomata:

a) COFC-F-5301; b) COFC-F-5305; c) COFC-F-5306. Scale bar = 1 cm.

free to almost free, with lamellulae, crowded, 8-12/cm, to 6 mm wide, white to

slightly pink [A1, 6A2] with concolourous margin; spore deposit pink [6A2].

ContTEXT solid, fibrous, <1 cm in pileus; odour sometimes of fresh fish (taste

not recorded). Termite mold ellipsoid, 10-20 cm long, grayish isabelline [4A5,

5C2] with irregular 0.5-1.5 cm wide holes.

MACROCHEMICAL REACTIONS — KOH- in pileus; HCl+ dark yellow in

pileus.

MICROCHARACTERS — PILEIPELLIS made up of doliiform to cylindrical

elements 6-17 x 3-9 um, hyaline with thin wall, somewhat constricted at

the septa; hyphal width increasing through context; parallel to interwoven

forming a lax ixocutis-subtrichodermis. PSsEUDORHIZzOID formed by parallel

cylindrical to doliiform elements, 4-15 um wide in the external zone with

often claviform to cylindrical terminal elements; hyphal width increasing

through context to 15-35 um wide, hyphae brown with thick wall, constricted

at the septa. STIPITIPELLIs similar to pseudorhizoid but hyphae hyaline, 3-5

um wide in the external zone and 7-18 um wide through context. CONTEXT

HYPHAE cylindrical, 9-21 um wide with thick 0.5-2 um wall, and mixed with

cylindrical hyphae 4-7 um wide, with thin wall. CLamp CONNECTIONS absent

346 ... Daniéls & al.

QanaanQn0avoals

OOO

PiateE 5. Termitomyces bulborhizus (COFC-F 5306):

a) Spores; b) Basidia; c) Cheilocystidia;

d) Pileus context; e) Pleurocystidia.

in all structures. CHEILocystTip1A claviform, fusiform to pyriform, (25-)

27-42 x (11-)13-24 um. PLEUROCYSTIDIA similar, sometimes with a secondary

septum and/or thick wall, 28-56 x (10-)11-20(-35) um. Basrp1a claviform,

18-24(-27) x 6-8.5 um, with (2-)4 sterigmata and thin to, rarely, thick-walled.

Spores ellipsoid, smooth, hyaline, thin-walled, (5.5-)6-7(-8) x (3-)3.8-4.4(-5)

um, [Lm = 6.4 um, Wm = 4.1 um, E = (1.38-)1.50-1.75(-1.80); Em = 1.56].

SPECIMENS EXAMINED — NIGER, Dosso, Gaya, Gaya, Gorou Bassounga Forest,

11°54’00’N 3°24’25”E, elevation 209 m, savannah, on termite nest, 5 September 2010,

O. Hama, Hama 443b (COFC-F 5302). TILLABERY, Say, Tamou, Haoussa, 12°15’18”N

2°22’14’E, elevation 221 m, wooded Afzelia africana Smith ex Pers. savannah, on

underground termite nest, 6 August 2010, O. Hama, Hama 286 (COFC-F 5301);

12°14’50’N 2°22’14”E, 24 August 2011, O. Hama, Hama 470 (COFC-F 5303); 25

August 2011, O. Hama, Hama 475 (COFC-F 5304); 12°14’51”N 2°22’15”E, wooded

savannah of Afzelia africana, Combretum glutinosum Perr. ex DC., C. collinum Fres.,

with Andropogon gayanus Kunth, Crossopteryx febrifuga (Afzel. ex G. Don) Benth.,

Strychnos spinosa Lam., and Flueggea virosa (Willd.) Royle, on active termite nest, 15

August 2012, O. Hama, Hama 519 (COFC-F 5305); Say, Torodi, Gnaktiré, 12°59’01”N

1°44’32”E, elevation 237 m, cropland, under Sclerocarya birrea (A. Rich.) Hochst., on

termite mound, 25 August 2010, O. Hama, Hama 405 (COFC-F 5095); 12°59’09.4”N

1°44’08.5”E, elevation 226 m, ina field of Pennisetum glaucum (L.) R. Br. and Sorghum

bicolor (L.) Moench, close to Balanites aegyptiaca (L.) Delile, on a termite nest, 19

August 2012, O. Hama, Hama 559 (COFC-F 5306).

EcoLoGcy & DISTRIBUTION — This taxon has been reported growing on

termite nests in southern China (Sichuan, Yunnan) and Thailand (Wei et al.

New Asian macromycete records for Africa ... 347

10 um

Pileipellis

10 um

Pseudorhiza

PLaTE 6. Termitomyces bulborhizus: comparison of pseudorhizoid and pileipellis

from exsiccata COFC-F 5306 (left) and isotype K 109284 (right).

2004; Sawhasan et al. 2011). The associated termite species is identified as

Macrotermes barneyi Light, native to Vietnam and southern China (Wang

et al. 2009) and Hypotermes makhamensis Ahmad in the Thailand collection

(Sawhasan et al. 2011). In this first African finding, the associated termite was

Macrotermes subhyalinus (Rambur) (Koné, pers. comm.).

ComMMENTS — Termitomyces bulborhizus is a large mushroom with a

characteristic black, swollen pseudorhizoid; the stipe surface appears slightly

scaly due to the presence of fine floccules.

Our material differs slightly from the isotype of T: bulborhizus (K 109284)

in the pileipellis structure that can be justified to different development stages

of the fruitbodies. Due to the few notes regarding pileipellis and pseudorhizoid

structure of this species, we consider showing them from both isotype and

COFC-F 5306 (PLATE 6). Descriptions of referred structures in the isotype are

as follows: pileipellis made up of doliiform to cylindrical elements 8-45(-78) x

4-17 um, hyaline with thin wall, somewhat constricted at the septa; hyphal width

increasing through context; parallel to interwoven forming a subtrichodermis

cutis; pseudorhizoid formed by parallel cylindrical to doliiform elements,

15-32 um wide, hyphae brown with thick wall, constricted at the septa. The

pseudorhizoid of the isotype was not clean enough for an accurate analysis so

348 ... Daniéls & al.

we do not know enough to ascertain whether it might also have small hairs or

claviform caulocystidia-like structures, as in the Nigerien sample. The bulbose

stipe width in Nigerien samples was also smaller (<3.5 cm wide) than those

given by Wei et al. (2004). Actually, Asian collections are bigger than African

ones regarding basidioma size, with a pileus of (5—)10-22 cm (Wei et al. 2004).

Our material differs from T! eurrhizus (Berk.) R. Heim in having a dark-

brown color and a swollen pseudorhizoid (Wei et al. 2004). In the most

similar African species (T. umkowaan (Cooke & Massee) D.A. Reid and

T. subumkowaan Mossebo) the stipe surface is smooth and the pseudorhizoid

is not swollen. Also, T. subumkowaan bears two spores per basidium (Mossebo

et al. 2002).

PLATE 7. Tulostoma evanescens (COFC-F 5292): Exsiccatum. Scale bar = 1 cm.

Tulostoma evanescens Long & S. Ahmad, Farlowia 3: 235, 1947. PLATES 7, 8

MACROCHARACTERS — BASIDIOMATA small, gregarious. GLOBOSE HEAD

0.7-0.9 cm diameter. ExoPERIDIUM undefined, hyphal, thin, brown [5B4],

intermixed with sand grains. ENDOPERIDIUM membranous, smooth, white [A1].

PERISTOME circular and not prominent, white [A1] because of the absence of

exoperidium around the mouth. StT1pe cylindrical or tapering upwards, 1-1.3

x 0.2-0.3 cm, surface squamose with evanescent scales and longitudinally

grooved, joined in the base to form a volva-like swollen structure in one

basidioma; sometimes with rhizomorphs. Socket lacerate, with several small

dentate membranes around the stipe. GLEBA brown coloured [6C6]. CONTEXT

firm, fibrous; odour absent and taste not recorded.

New Asian macromycete records for Africa ... 349

000000e0000 |:

; S

PLATE 8. Tulostoma evanescens (COFC-F 5292):

a) Exoperidium; b) Context of stipe scales; c) Endoperidium;

d) Hyphae of rhizomorph; e) Spores; f) Capillitium.

MICROCHARACTERS — EXOPERIDIUM with cylindrical interlaced hyphae,

1.5-4 um wide with, hyaline to brownish, moderate wall. ENDOPERIDIUM

similar, with hyphae 3-6(-8) um wide, hyaline and with thick wall.

CAPILLITIUM 3-8 tm wide, with lumen and thick wall minutely encrusted with

granules, septate and with a few branches. HYPHAE OF STIPE SCALES parallel,

4-6.5 um wide, grouped in tufts; hyphae hyaline to yellowish, with thin wall,

somewhat constricted at the septa. HYPHAE OF RHIZOMORPH 2.5-7 um wide,

hyaline, often minutely encrusted with granules and with moderate wall.

350 ... Daniéls & al.

CLAMP CONNECTIONS absent in all structures. BAsIDIA not seen. SPORES ovoid

to ellipsoid, smooth, brown coloured, thick-walled, some apiculate, (4.7-)

5-5.4(-6) x (3.7-)4-4.5(-5) um, [Lm = 5.2 um, Wm = 4.1 um, E = 1.11-1.38;

Em=126].

SPECIMEN EXAMINED — NIGER, TILLABERY, Say, Tamou, Tamou Total Faunal Reserve

{Tamou Reserve}, 12°34’21”N 2°18’28”E, elevation 240 m, wooded savannah, humus-

containing sandy soil, 7 August 2010, O. Hama, Hama 361 (COFC-F 5292).

ECOLOGY & DISTRIBUTION — ‘This species grows in arid sites and appears

after rains in copious quantities. According to Wright (1987), this species has

been reported in India and Argentina. This appears to be the first report of this

species in Africa.

ComMENTs — The material examined here agrees with earlier descriptions,

including the following features: 1) Clearly-defined circular, tubular, or

flat peristome on a head of <1 cm in diameter; 2) smooth subglobose or

ellipsoid spores measuring about 5-5.4 x 4-4.5 um; 3) inconspicuous hyphal

exoperidium; and 4) capillitium 4-10 um wide.

Tulostoma operculatum Long & S. Ahmad has smaller (4-4.7 um diam.)

spores and a fimbriate mouth. Tulostoma brevistipitatum B. Liu et al. does not

havea volvoid structure at the stipe base, and its spores are slightly longer (5.4-6.1

x 4.3-5 um). Tulostoma fusipes Har. & Pat. is very similar, but its spores are

globose, the capillitium is coloured, and the stipe is longer (5.5 x 0.4 cm)

(Wright 1987).

Volvariella cf. sathei Senthil., Rahul Sharma & S.K. Singh, Mycotaxon 119: 470,

2012: PLATES 9, 10

MACROCHARACTERS — BASIDIOMATA medium sized, gregarious. PILEUS

3.5-12(-15) cm diameter, fleshy, convex; white to isabella [A1l, 4A2]; surface

fibrillose; margin exceeding the lamellae, eroded, or striate with triangular

tufts. Stipe central, fistulose when aged, cylindrical and tapering towards

apex, straight, concolourous with pileus, 5-12 x 1-2 cm, surface glabrous to

fibrillose; slightly bulbose. VoLva membranous, 1-5 x 2-4 cm and 1-1.5 mm

thick, white [A1], sometimes with grayish brown hue [4C3]. LAMELLAE free,

distant from stipe, with lamellulae, crowded, (10-)13-16(-17)/cm, 3-5 mm

wide, at first whitish [Al, 3A2] then pink [9A2, 14A3] with paler, dentate to

fibrillose margin. Spore deposit pink [9A3, 10B5]. CONTEXT solid, fibrous;

odour disagreeable, farinaceous to fruity, and pleasant taste.

MACROCHEMICAL REACTIONS — KOH- or NaOH- in pileus, context or

stipe.

MICROCHARACTERS —PILEIPELLIS made up of cylindrical elements

8-21 um, hyaline with thin to thick wall, somewhat constricted at the septa;

parallel, forming a regular cutis. STIPITIPELLIS similar to pileipellis, hyphae

New Asian macromycete records for Africa ... 351

PLATE 9. Volvariella cf. sathei, fresh basidiomata:

a) COFC-F-5092; b) COFC-F-5036; c) COFC-F-5033. Scale bar = 3 cm.

hyaline, 4-15 um wide in the external zone and up to 35 um wide through

context. CONTEXT HYPHAE cylindrical, 8-21(-28) um wide with thin wall.

VoLvaA composed mostly of cylindrical hyphae, 3-5 um wide with scattered

inflated elements, cylindrical, fusiform or irregularly shaped, up to 20 um wide.

BASAL TOMENTUM formed by interlaced cylindrical elements, 5-22 um wide;

hyaline with thin to thick wall. CLamp CONNECTIONS absent in all structures.

CHEILOCYSTIDIA fusiform to lageniform, 40-88(-95) x 10-38 um, with

hyaline thin wall, sometimes with a basal secondary septum. PLEUROCYSTIDIA

similar, 39-80 x 11-24 um. SUBHYMENIUM cellular, with ellipsoid to doliiform

hyphae, 10-20 um in diameter. Basrp1a claviform, (19-)22-27(-30) x 7-9

um, with (2—)4 sterigmata and thin-walled. Spores globose to ob-triangular,

smooth, brownish pink, thick-walled, (5-)5.6-6(-7) x (4.5-)5.1-5.7(-6.7) um,

[Lm = 5.8 um, Wm = 5.4 um, E = 0.92-1.40; Em = 1.07].

SPECIMENS EXAMINED: NIGER, NiaMeEy, Niamey, Abdou Moumouni University,

Faculty of Agricultural Sciences, experimental garden, 13°30'00.5”N 2°05’24.5”E,

elevation 178 m, in a disturbed area, growing on plant residue under Prosopis juliflora

(Sw.) DC., 6 August 2009, O. Hama, det. A. Justo, Hama 175 (COFC-F 5036); sandy

352 ... Daniéls & al.

99900000000] E

g ie eal

PiatE 10. Volvariella cf. sathei (Fics a-e, g—h from COFC-F 5448; Fia. f from COFC-F 5033):

a) Pileus context; b) Pileipellis; c) Spores; d) Basidia; e) Cheilocystidia;

f) Volva, g) Basal tomentum, h) Stipitipellis.

soil with cattle manure under Prosopis juliflora, O. Hama, Hama 176 (COFC-F

5092); Niamey, Abdou Moumouni University, High School of Education, 13°30’07”N

2°05’29’E, elevation 187 m, in a disturbed area, in sandy soil near Azadirachta indica

A. Juss., 4 August 2010, O. Hama, det. A. Justo, Hama 390 (COFC-F 5088; Genbank

KF926666). TILLABERY, Say, Torodi, Fayra, 13°01’26.7”N 1°45’39.4’E, elevation 232 m,

in arable fields with plant residue of Piliostigma reticulatum (DC.) Hochst., 21 August

New Asian macromycete records for Africa ... 353

2008, O. Hama, Hama 38 (COFC-F 5448); Say, Torodi, Gnaktiré, 12°59’02”N 1°44’49’E,

elevation 222 m, cropland, growing on organic residue near houses with Pennisetum

glaucum and Sorghum bicolor, 16 August 2009, O. Hama, det. A. Justo, Hama 194

(COFC-F 5034; Genbank KF926664); 12°59’01”N 1°44’32”E, elevation 221 m, in arable

fields on heaped-up plant residue with Sorghum and Pennisetum, 15 August 2009, O.

Hama, det. A. Justo, Hama 193 (COFC-F 5035; Genbank KF926663); Say, Tamou,

Mékrou, {W National Park}, 12°15’16”N 2°23’24’E, elevation 218 m, in gallery forests

with Cola laurifolia Mast., Diospyros mespiliformis Hochst. ex A. DC., Mitragyna inermis

(Willd.) Kuntze, on sandy-clay soils, 22 August 2009, O. Hama, Hama 204, det. A. Justo

(COFC-F 5033; Genbank KF926665).

EcoLoGy & DISTRIBUTION — ‘The authentic species, Volvariella sathei, was

recently proposed, based on Indian material (Senthilarasu et al. 2012), and has

not been reported elsewhere.

ComMENTsS — The large white species, V. sathei, differs from V. nivea 'T.H. Li

& Xiang L. Chen in having ovoid to widely ellipsoid spores and larger cystidia

(Senthilarasu et al. 2012).

Our samples agree mostly with the description given by Senthilarasu et al.

(2012) except for: (1) the presence of fusiform to lageniform cystidia (cylindro-

clavate in V. sathei); (2) spores often with a triangular shape; and (3) grayish

hue sometimes present in volva. Molecular analysis indicates that the African

isolates are in the same clade as the Indian V. sathei but that there are some

differences between them (PLATE 11). Additional data are needed to confirm

whether the African isolates represent V. sathei or a closely related taxon.

89°>- HM246492 Volvariella hypopithys TOAV137 Sardinia

'— JF415139 Volvariella sp LOU18924 Spain

af HM246494 Volvariella pusilla Sardinia TOAV139

100 JF415137 Volvariella pusilla AJ51 Spain

98 -—— HM246493 Volvariella strangulata Italy TOAV141

'—— HM562213 Volvariella surrecta AJ55 Spain

95 '—— JF415141 Volvariella terrea LUG 11010 Holotypus France

JF415140 Volvariella dunensis SCM3513 Spain

100 JF415138 Volvariella dunensis RFS 07031003 Spain

| JF415136 Volvariella dunensis TOAV140 Sardinia

86 HM 246491 Volvariella taylorii TOAV142 Sardinia

HM562214 Volvariella lepiotospora AJ155 USA

0.06 |_| -___________ 562211 Volvariella caesiotincta MA54717 Spain

HM246500 Volvariella sp TOAV143 Slovenia

HM562210 Volvariella taylorii AJ54 Portugal

g3)KF 926664 Volvariella cf. sathei Hama194 Niger

KF926663 Volvariella cf. sathei Hama193 Niger

KF926665 Volvariella cf. sathei Hama204 Niger

| erszeee Volvariella cf. sathei Hama390 Niger

JN792550 Volvariella sathei AMH 9436 India

HM562212 Volvariella bombycina AJ244 Spain

FJ379274 Volvariella volvacea VV34

PLaTE 11. Best tree from the Maximum Likelihood analysis of ITS sequences of Volvariella.

Nigerien samples of V. cf. sathei are emphasized. Scale bar indicates nucleotide substitutions/site.

100

354 ... Daniéls & al.

Discussion

The taxa examined here are species previously recorded in Asia; this is the

first time they have been reported on the African continent. Fungal species

common to both continents are frequently reported, perhaps due to climatic

and/or ecological similarities, spore dispersal capacity, and shared geological

history; cases of vicariance and species fragmentation are also found (Berndt

2002, O'Donnell et al. 2011, Geml et al. 2008, Moncalvo & Buchanan 2008).

The fragmentation of biogeographical ranges may sometimes be due to human

intervention and particularly to the introduction of exotic plant species with

their own associated fungal suites (Niveiro et al. 2009, Chen et al. 2006, Read

2000, Pringle et al. 2009); some ectomycorrhizal fungi entered Africa by this

means (Duponnois & Galiana 2007, Garbaye et al. 1988). The climate of the

West Sudanian savannah is similar to that of the Asian savannah of India,

Laos, Thailand, Myanmar, Vietnam, and Cambodia (Young & Solbrig 1993).

There are, however, differences in floral composition, with Dipterocarpaceae

predominating in Asia and Leguminosae in Africa. This can help to discriminate

some tree-restricted ectomycorrhizal species better than saprobes. However,

there are some wide-ranging mycorrhizal fungi, which could also suggest a

lack of fungal sampling in this ecosystem type (Tulostoma evanescens might

represent such a case). Actually, infrequent sampling of the African savannah

may have led to a bias in our knowledge of the biogeographical range, substrate

versatility, spore dispersal capacity, and ecological similarity of these species.

Furthermore, some saprobic species are often unspecific regarding substrate

degradation and sometimes the substrate itself is originated by fire or domestic

herbivore digestion, reducing substrate differences between distant geographic

areas.

Mushrooms associated with termite nests grow only where termites are

found. Several termite genera and species (e.g. Macrotermes, Microtermes,

and Odontotermes) are common to both continents (Cheng and al. 2011) and

this kind of mycological association seems to have originated in West Africa

(Nobré et al. 2011). Clitopilus orientalis is a saprobic fungus with an apparent

preference for termite nests.

Our discoveries provide a promising basis for further research into Niger’s

fungal diversity. Many collections are yet to be reviewed, and their examination

will doubtless enhance our knowledge of fungal diversity in the West Sudanian

savannah as a whole, and also enable a closer appraisal of its similarity with its

Asian counterpart. More samples and DNA-analysis of the studied material are

needed to discern cryptic species, and Volvariella cf. sathei may be a geographic

variety of the Asian species.

New Asian macromycete records for Africa... 355

Acknowledgments

This study was part of the “Edible and cultivable Macromycetes of Niger

(Ethnomycology)” project, financed by the Spanish Agency for International

Development Cooperation (AECID-C/023163/09; D/031488/10; A1/039675/11). The

authors are grateful to the curators of the COFC, K and BR herbaria for the exsiccata

loan management. They would also like to thank the mycologists T.J. Baroni (State

University New York, Cortland) and L.A. Parra (Aranda de Duero) for providing

assistance, comments, and taxonomic information regarding the identification of

certain species; also thanks to Nourou Soulemane Yorou (University of Miinchen)

and Gabriel Moreno (University of Alcala de Henares) for their critical revision of the

manuscript. We are indebted to N’Golo Abdoulaye Koné (University of Abobo-Adjamé)

for the identification of Macrotermes subhyalinus and to M. Robichaud (Ottawa) for the

English revision of the manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/130.361

Volume 130, pp. 361-367 April-June 2015

Contribution to the knowledge of Inonotus baumii

in Thailand

ANON AUETRAGUL’, ORATAI EUATRAKOOL?’,

MarIA LETIZIA GARGANO}, & GIUSEPPE VENTURELLA?

' AnonBiotec Institute, Department of Biotech, The North Chiangmai University,

Talad Thai, Klongsong, Klongluang, Patumthani 12120, Chiangmai, Thailand

? Office of Agricultural Regulation, Department of Agriculture,

Chatuchak, Bangkok, 10900 (Thailand)

° Department of Agricultural and Forest Sciences, University of Palermo,

Viale delle Scienze, Bld. 4, I-90128 Palermo (Italy)

* CORRESPONDENCE TO: giuseppe.venturella@unipa. it

ABSTRACT —Inonotus baumii in Thailand is reported on three new host-plants: Acacia

tomentosa, Shorea robusta, and S. siamensis. A key for the identification of Inonotus species in

Thailand accompanies notes on the taxonomy of I. baumii.

Key worps — Basidiomycota, Hymenochaetales, Phellinus, white rot fungus

Introduction

Fungi are one of the most species-rich groups of organisms but compared to

plants they are still vastly under-investigated (Hawksworth 1997). In particular

there is still a lack of distribution and ecological data on several fungal species

(Venturella et al. 2011). This is even more true for wood-inhabiting fungi,

which have become increasingly important in recent years not only for their

ecological role but also as natural sources of anticancer, antioxidative, and

antibacterial agents (Karaman et al. 2010).

Several wood-decaying species are used in Asian folk medicine (Larsson

et al. 2006). An example is Inonotus baumii, sometimes erroneously named

Phellinus linteus (Berk. & M.A. Curtis) Teng, which is known for its use in

traditional Chinese medicine (Ying et al. 1987, Shon et al. 2003). Larsson et al.

(2006) have researched the perplexing taxonomy of Hymenochaetales, while

Tura et al. (2011) have conducted cultural-morphological investigations on

362 ... Auetragul & al.

I. baumii [sub Phellinus baumii] and P. linteus. According to these authors,

the taxonomic delimitation at the generic and species level in Phellinus s.l. and

Inonotus s.l. is confusing, and P. linteus and I. baumii [sub P. baumii] show

close phylogenetic relationships within the P. linteus species complex.

Wu et al. (2012) investigated the species delimitation of the I. baumii-

I. linteus group (including taxa recorded as P. linteus) and included the

“sanguangh mushroom, a popular polypore used medicinally in China,

Japan, and Korea. They described the “sanguangh mushroom” as a new species

(Inonotus sanghuang Sheng H. Wu et al.) growing solely on Morus sp. in China,

Japan, Korea, and Taiwan. Wu et al. (2012) also reported I. baumii on living

Syringa sp. distributed in temperate NE Asia. Parmasto & Parmasto (2001)

highlighted the need for correctly identifying the host species when collecting

Phellinus species on trees or bushes in tropical and subtropical areas. The effects

of Cambodian P linteus in the wood chemistry and structure of Shorea obtusa

Wall. ex Blume were investigated by Srivilai et al. (2013).

The aim of this paper is to contribute to the knowledge of Inonotus baumii

in Thailand and to report three new host plants.

Materials & methods

The studied basidiomata were collected in different periods of the year in Chiangmai

(Mai Hongson Province) and the northeastern region of Thailand; the morphological

characters of basidiomata kept in the personal collection of one of the authors

(A. Auetragul) were also examined. The morphology of fresh and dried basidiomata

was analyzed in the Laboratory of Mycology of the Department of Agricultural and

Forest Sciences (University of Palermo, Italy). The basidiospore length and width, mean

number of pores per mm, mean size of setae, and diameter of tramal skeletal hyphae

were determined according to Parmasto & Parmasto (2001). Morphological characters

were also evaluated according to the identification keys of Wu et al. (2012) and Tian

et al. (2013), which consider the hymenial surface pore sizes, basidiospore length and

width, basidioma type, pileus surface characters, color of the pileal surface marginal

zone, and host-plants. The host-plants on which we collected the basidiomata were

identified according to Graf (1992). Fungal nomenclature follows MycoBank. The dried

herbarium specimens are kept in the collection of AnonBiotec Institute, Department

of Biotech, The North Chiangmai University, Thailand (DBNC) and the Herbarium,

Department of Agricultural and Forest Sciences, University of Palermo, Italy (SAF).

Taxonomy

Inonotus baumii (Pilat) T. Wagner & M. Fisch., Mycologia 94: 1009 (2002) PLaTE 1

Basidiomata woody, perennial, sessile, semicircular, ungulate or imbricate,

7 x 15 cm, 5-6 cm thick at the base, pileal surface dark grayish brown to

dark gray, densely sulcate, radially rimose and cracking; margin rounded;

hymenophores golden yellow when unripe then dark brown with wide and

Inonotus baumii in Thailand ... 363

PLATE 1. Inonotus baumii basidiomata (photos courtesy of AnonBiotec).

A, from Shorea robusta (DBNC 021, SAF 038); B. from Acacia tomentosa (DBNC 020).

sterile zone along the margin; pores regular, circular 8-10 per mm; tube layers

receding in old basidiomata; context woody. Hyphal system dimitic; generative

hyphae simple-septate, hyaline to golden yellow, 2-2.5 um wide; skeletal

hyphae golden yellow to golden ferrugineous, thick-walled, up to 4-5 um wide;

hymenial setae frequent to scanty, ventricose with an acute apex, thick-walled

at the base, 20-27 x 5-13 Um; basidia clavate, 7-12 x 5-8 um, tetrasporic;

basidiospores broadly ellipsoid to subglobose, hyaline then pale yellowish

brown, thin-walled, 6 x 5 um.

SPECIMENS EXAMINED: THAILAND. NORTHERN THAILAND: CHIANG MAI PROVINCE,

Chiang Mai, UTM 2077469 498244 47Q, 310 m, on living and dead trees of Acacia

tomentosa Willd. (Leguminosae), 20 Oct 2012, A. Auetragul (DBNC 020); Maz Hone

SON ProvincE, Mae Hong Son, UTM 2134467 391785 47Q, 240 m, on living and dead

trees of Shorea robusta C.F. Gaertn. (Dipterocarpaceae), 17 Oct 2013, A. Auetragul

(DBNC 021, SAF 038); LaMpaNG Province, Lampang, UTM 552842 2023449 47Q, 244

m, on living and dead trees of Shorea siamensis Miq. (Dipterocarpaceae), 25 Oct 2013,

A. Auetragul (DBNC 022).

Identification key for Inonotus spp. in Thailand

(partly modified from Wu et al. 2012)

bPorés or hymenial Surface 5/ DANG tee tee eee cote eon «eee I. lonicerinus

LpPoresiol- hymemial-siintace! S5/ Ini .a5 Bae wtt Boe Ae AE ae hin eee ae Ai Bi 2

2. On dead branch of Lonicera spp. Basidiospores mostly <4. 1 x 3.1 um

I. lonicericola

2. Not on Lonicera spp. Basidiospores mostly >4.1 X 3.LUMm.............. 0... eee 3

3. On fallen trunks of Populus spp. Basidiomata resupinate or pileate. Pileus surface

indistinctly or moderately sulcate with furrows <3/cm. Pileus always with a wide

yellowsmarginalzone onthe surtace ss: bu 5 aes es ae cae I. vaninii

3. Not on Populus spp. Basidiomata pileate. Pileus surface densely sulcate with furrows

>3/cm. Pileus usually lacking a wide yellow marginal zone on the surface in ripe

basidioniata: (Asc. tins -ehines Hein aioe eRince tein? Mme + eee + elds om taht om, 4

364 ... Auetragul & al.

4. On Weigela coraeensis. Basidiomata sessile to effused-reflexed. Pileus applanate,

frequently becoming decurrent with more or less effused base, surface with a

thin and black crust, margin usually thin and acute................ I. weigelae

4. Not on Weigela. Basidiomata always sessile. Pileus applanate to concave or

subungulate; surface without a thin crust. Pileus surface sometimes black and

crustose in ripe basidiomata. Pileus margin acute or blunt ................... 5

5. On Morus spp. Basidiomata perennial, sessile, pileate. Pileus applanate or applanate

with an umbo to convex, adaxially flat, slightly convex or slightly concave. Pore

surface golden-yellow, brownish yellow to yellowish brown. Pileus margin

turmine-dark=redewithyi@ Flt ie: he biG aphasia aa I. sanghuang

5. On dead and living trees of Syringa sp., Acacia tomentosa, Shorea robusta, S.

siamensis. Basidiomata always sessile, applanate to subungulate, adaxially more

or less concave, margin acute or blunt, thin crust absent, although pileus surface

may become black and crustose in ripe basidiomata, surface densely sulcate,

with farrows >3/cm. Aged pileus usually lacking a wide yellow marginal zone

on pileus surface. Pileus margin not turning dark-red with KOH. Pore surface

yellowish brown. Pores of hymenial surface >5/mm. Basidiospores more or less

sphiaerical,6im long:and Siti wide ie . casege seeing eae) ees wee I. baumii

Inonotus baumii (Hymenochaetaceae) belongs to the I. baumii-I. linteus group

distributed in temperate NE Asia. It has been recorded on living and dead

trees of Syringa (Wu et al. 2012) in China (Dai 2010), Japan (Ito 1955), Korea

(Lim et al. 2003), and Far East Russia (Parmasto & Parmasto 2001). Phellinus

linteus has been reported to grow on Quercus L., Cassia L., Lonicera L., Morus

L., Symplocos Jacq., and Hemiptelea Planch. (Ahmad 1972, Kang et al. 2002,

Larsen & Cobb-Poulle 1990, Lim et al. 2003). However, Lim et al. (2003), who

investigated basidiomata from China, Korea, Costa Rica and, Mexico, showed

that the temperate East Asian species known as “P. linteus” represents I. baumii.

Wt et al. (2012) proved that specimens collected on Morus species represent

another taxon, I. sanghuang.

Discussion

The difficulties in identifying Inonotus and Phellinus spp. belonging to the

P. linteus species complex fuel a wide debate on the taxonomic status of the

collected basidiomata growing on different wild and ornamental trees. This is

also due to the great interest by scientists and people towards the medicinal

use of the mushrooms identified as P. linteus and/or I. baumii. The worldwide

scientific movement on medicinal mushrooms and the mushroom industry

(mainly in Asian countries) specifically requires an exact identification of

mushroom species (Wasser 2013). According to our investigation the host

range of the P. linteus species complex is not exhaustive. We report Inonotus

baumii for the first time on three new host plants growing in Thailand: Acacia

tomentosa (PLATE 2), Shorea robusta (PLATE 2), and S. siamensis (PLATE 3).

Inonotus baumii in Thailand ... 365

PLATE 2. (A) Acacia tomentosa in Chang Mai (Thailand);

(B) Shorea robusta in Mae Hong Son.

PiaTE 3. Shorea siamensis in Lampang (Thailand).

366 ... Auetragul & al.

These plants are cultivated as ornamentals in Chiang Mai, Mae Hong Son,

and also in Lampang. Depending on its host, different vernacular names are

attributed to I. baumii in Thailand: the local name of Acacia tomentosa is

“Kra-Tin-Piman” and I. baumii on this host is called “Hed Kra-Tin-Piman”

(Hed = mushroom); the local name of Shorea robusta is “Jigg” (North Thailand)

or “Keng” (North-Eastern Thailand) so I. baumii on this host is known as “Hed

Jigg” or “Hed Keng”. Given that I. baumii is collected by a large number of

people attracted by the medicinal properties of the basidiomata, the above

identification key should enable non-specialists also to identity I. baumii.

Acknowledgments

The authors wish to thank Prof. Georgios Zervakis (Greece) and Prof. Omon

Isikhuemhen (USA) for carefully reading the manuscript and for English revision.

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MYCOTAXON

http://dx.doi.org/10.5248/130.369

Volume 130, pp. 369-397 April-June 2015

Taxonomic studies in Chrysoderma, Corneromyces,

Dendrophysellum, Hyphoradulum, and Mycobonia

KAREN K. NAKASONE

Center for Forest Mycology Research, U.S. Forest Service

One Gifford Pinchot Drive, Madison, WI 53726 USA

*CORRESPONDENCE TO: knakasone@fs.fed.us

ABSTRACT — Eight poorly known or unusual crustose and pileate basidiomycete species

were studied. These included the type specimens of three monotypic genera: Chrysoderma

alboluteum from Réunion is conspecific with Cerocorticium molle; Dendrophysellum

amurense from the Russian Far East is a species of Vararia; and Hyphoradulum conspicuum

belongs in Pseudolagarobasidium and is the first representative of the genus from Europe.

Corticium murrillii, from Mexico, is congeneric with Corneromyces kinabalui. New

combinations Vararia amurensis, Pseudolagarobasidium conspicuum, and Corneromyces

murrillii are proposed. Mycobonia flava and M. brunneoleuca are macroscopically similar

species with diagnostically distinct basidiospore shape and size. Mycobonia disciformis is

accepted in Mycothele, and Mycobonia winkleri represents a species of unknown affinities.

Recent molecular phylogenetic studies indicate that Mycobonia is embedded in Polyporus

sensu stricto and is a synonym of Polyporus. Transferring M. brunneoleuca and M. flava

to Polyporus requires the creation of the replacement names, P. polyacanthophorus and

P. epitheloides.

Key worps — Amylocorticiales, cyanophilous basidiospores, Epithele, Favolus curtipes,

Polyporales

Introduction

Tremendous advances in the systematics of basidiomycetes have been

made in the last twenty years since the advent of molecular phylogenetics.

Nevertheless, morphological studies are still essential to advance this discipline.

In this paper, eight species of poorly known crustose or pileate basidiomycetes

are described and illustrated. Type specimens of Chrysoderma alboluteum,

Corticium murrillii, _Dendrophysellum amurense, and Hyphoradulum

conspicuum were examined. In addition, the genus Mycobonia (Polyporales)

is revisited. Species of Mycobonia produce stipitate, pileate basidiomes with

hyphal pegs penetrating a smooth hymenial surface. Four species have been

370 ... Nakasone

included in Mycobonia — M. brunneoleuca, M. disciformis, M. flava, and

M. winkleri (Julich 1976). Mycobonia brunneoleuca is considered a synonym of

M. flava by some authors, and M. disciformis was transferred to the monotypic

genus Mycothele by Jilich (1976). Little is known about M. winkleri, which has

not been studied since it was first described in 1911. In this paper, Mycobonia

brunneoleuca and M. flava are accepted as distinct species and transferred to

Polyporus Adans. The literature concerning M. flava and M. brunneoleuca is

critically examined. All the taxa are described and illustrated.

Materials & methods

Thin, freehand sections from basidiomes were mounted in Melzer’s reagent (Kirk et

al. 2008) or in 1% (weight/volume) aqueous phloxine and 2% (w/v) aqueous potassium

hydroxide. Cyanophily of basidiospore and hyphal walls was observed in 0.1% cotton

blue in 60% lactic acid (Kotlaba & Pouzar 1964; Singer 1986). Basidiome sections were

mounted in freshly prepared sulfovanillin solution (1 g vanillin, 3 ml distilled water,

8 ml concentrated sulfuric acid); a positive reaction was recorded if contents of the

cystidium turned black. Drawings were made with a camera lucida attachment on an

Olympus BH2 compound microscope. Q values were obtained from dividing average

basidiospore length by width (Kirk et al. 2008). Basidiospores are sometimes scarce

in specimens, thus Q values based on less than 30 basidiospores are approximate and

indicated with an asterisk (*). Color codes are from Kornerup & Wanscher (1978) except

that capitalized color names follow Ridgway (1912). Herbarium code designations

follow Index Herbariorum (Thiers 2014). Accepted species names are in boldface.

Taxonomy & discussion

Chrysoderma alboluteum Boidin & Gilles, Cryptog. Mycol. 12: 127. 1991.

FIGS 1-2, 10-11

= Corticium molle Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 336. 1868 [“1869”].

= Cerocorticium molle (Berk. & M.A. Curtis) Jiilich, Persoonia 8: 219. 1975.

= Corticium armeniacum Sacc., Syll. fung. 6: 637. 1888.

= Terana armeniaca (Sacc.) Kuntze, Revis. gen. pl. 2: 872. 1891.

= Corticium ceraceum Berk. & Ravenel, in Massee, J. Linn. Soc., Bot. 27: 150. 1890 [“1891”].

= Cerocorticium bogoriense Henn. & E. Nyman, Monsunia 1: 139. 1900 [“1899”].

= Cerocorticium tjibodense Henn., Monsunia 1: 139. 1900 [“1899”].

= Corticium aureolum Bres., Ann. Mycol. 9: 272. 1911.

BASIDIOME resupinate, widely effused, orbicular at first then confluent, <10

x 3 cm, rarely <1 m long, often breaking up into smaller pieces when dried,

often slightly detached from substrate, <650 um thick, ceraceous to corneous,

smooth to slightly warted, white at first, then dull yellow, light orange (5A4),

brownish orange (6C7), brown [7D(6-7)], Warm Buff, Russet, Tawny, Hazel, or

Snuff Brown, sometimes slightly cracked on drying. CONTEXT white to cream-

colored. MARGIN distinct, abrupt, edges detached or curling slightly away from

Unusual crust and pileate basidiomycetes ... 371

substrate, thickly fibrillose, or thinning out, concolorous with hymenium,

white, or dark yellow.

HYPHAL SYSTEM monomitic with clamped generative hyphae. SuBicuLUM

<400 um thick, a moderately dense, non-agglutinated tissue of more or less

vertical hyphae; subicular hyphae (2-)3-6 um diam, clamped, moderately

branched, walls hyaline, thin to 1.5 um thick, smooth, acyanophilous.

SUBHYMENIUM <75 um thick, a dense, compact but non-agglutinated tissue

of vertically arranged hyphae; subhymenial hyphae 4-5.5 um diam, clamped,

frequently branched, walls hyaline, thin, smooth. HyMENtum <100 um thick, a

dense, non-agglutinated palisade of hyphidia and basidia sometimes embedded

in mucilaginous material HypuHip1A sometimes absent, when present

filamentous with slight constrictions or swellings, sometimes with knobby

outgrowths at apex, 35-70 x 2-4 um diam, clamped at base, rarely branched,

walls hyaline, thin or sometimes basally thickened, smooth. Basip1a narrowly

clavate with a long stalk, (40-)65-85(-100) x 7.5-10(-12) um, clamped at base,

often containing oil-like inclusions, walls hyaline, thin to slightly thickened,

smooth, 4-sterigmate, sterigmata 7-10 x 1.5-2.2 um. BASIDIOSPORES

scarce to numerous, narrowly cylindrical to cylindrical, (13-)15-20(-23) x

4.5-8(-9) um, average of five specimens 16.4-19.4 x 5.7-7.9, Q = (2.1-)2.6-3.1,

often containing oil-like, cyanophilous materials, walls hyaline, thin, smooth,

acyanophilous, not reacting in Melzer’s reagent.

HABITAT & DISTRIBUTION — On bark and wood of various dead

angiosperms, occasionally on or under bark of living trees, throughout tropical

and subtropical areas of North America, South America, Asia, and Africa.

TYPE SPECIMENS EXAMINED — REUNION. Anse des Cascades, sur branche tombée au

bordure de leau, 6 avril 1990, J Boidin (LY 14412, holotype of C. alboluteum). UNITED

STATES. SoutH Caro ina: ad corticeum ramulosque Corni, 1855, H Ravenel — Fungi

Caroliniana, Fasc. III no. 29 (BPI 280736, 280735, syntypes of C. ceraceum).

ADDITIONAL SPECIMENS EXAMINED — BRAZIL. Parana: Curitiba, Capao da Imbuia,

on decayed, dicot trunk, 16 Nov 1992, AAR de Meijer 2367 (CFMR). SAo LEOPOLDO:

in ligno frondoso, 1907, Rick - Fungi Austro-Americani no. 240, as Aleurodiscus

alboroseus Bres. (BPI 280723, as C. ceraceum). MEXICO. Orizaba, Rincon Grande

and El Barrio, 4000 ft, on decorticate branch, 10-14 Jan 1910, WA & EL Murrill 54615

(BPI 280733, as C. ceraceum). PUERTO RICO. LuquILLo Municipio: Sabana barrio,

ridge above chicken farm along Rio Sabana, on decorticated hardwood, 30 Jun 1996,

KK Nakasone (CFMR FP 150011). REUNION. St. Gilles I, en partie vivante Casuarina

equisetifolia L. (bark), 26 avril 1985, J Boidin (LY 11356, as C. alboluteum); St. Gilles II,

ravine, sur branche au sol, 26 avril 1985, J Boidin (LY 11373, as C. alboluteum). SOUTH

AFRICA. No location, on bark, no date, P van der Bijl 13 (BPI 280731). UNITED

STATES. FLoripa: Highlands Hammock, on bark of Quercus sp., 8 Feb 1937, CL Shear

330 (BPI 28038). Loutsi1Ana: Lafayette Parish, on rotten Cornus florida L., 7 May 1887,

AB Langlois 1467 (BPI 289279, as C. armeniacum); St. Martinsville, on bark, 21 Apr

1897, AB Langlois 2389 (BPI 330776, as C. ceraceum). SOUTH CAROLINA: on Cornus sp.,

no date, H Ravenel, Ellis — North American Fungi no. 607 (BPI 280278).

372 ... Nakasone

Bd ?

: ie 3) ; F : >

Ki be z

Fics 1-9. Cerocorticium molle (FP 150011). 1. Basidiome. Chrysoderma alboluteum (LY 11356).

2. Basidiome. Corticium murrillii (isotype HUH 00290581). 3. Basidiome surface; 4. close-up of

basidiome surface; 5. subicular hyphae in Melzer’s reagent; 6. basidiospore with cyanophilous walls;

7. basidiospore with bluish black, amyloid walls in Melzer’s reagent. Dendrophysellum amurense

(holotype TAAM 015561). 8. Basidiome; 9. close-up of hymenial surface. Scale bars: 1, 8 = 10 mm;

2= 2 mm; 3, 4,9 = 1 mm; 5, 6 = 10 um.

Unusual crust and pileate basidiomycetes ... 373

FiGs 10-11. Corticium ceraceum (syntype BPI 0280736). 10. A. Basidiospores; B. basidia;

C. hyphidia. Cerocorticium molle. 11. Basidiospores: A. from BPI 0280731; B. from BPI 0280733.

DESCRIPTIONS & ILLUSTRATIONS — Boidin & Gilles (1991, as C. alboluteum), Burt (1926:

216, as C. ceraceum), Hjortstam (1983), Hohnel & Litschauer (1907, as C. ceraceum),

Julich (1975), Maekawa et al. (2003), Nakasone (2008, as C. aureolum), Talbot (1951, as

C. armeniacum), Trierveiler-Pereira et al. (2009), Wu & Chen (1990).

COMMENTS — Cerocorticium molle is characterized by yellow, orange, or

brown, ceraceous to corneous basidiomes, simple hyphidia, large basidia, and

large, cylindrical to narrowly cylindrical basidiospores. The basidiospores are

variable in shape and size, sometimes within a single specimen. Hohnel’s (1910)

and Julich’s (1975) synonymies of Cerocorticium bogoriense and C. tjibodense,

both from Java, with C. molle are accepted.

374 ... Nakasone

There is no doubt that C. alboluteum is conspecific with C. molle as suggested

by Hjortstam & Larsson (1995). Although hyphidia were not observed in

specimens of C. alboluteum examined, basidiomes and basidiospores are

typical for the species. Boidin & Gilles (1991) noted that in C. alboluteum

the basidiospore walls were pale yellow, producing a mass basidiospore color

of yellowish orange, and calculated the average basidiospore size of three

specimens as 12.8-14.6 x 4.4—5.1 um, Q = 2.8-3.

Corticium murrillii Burt, Ann. Missouri Bot. Gard. 13: 289. 1926. FIGS 3-7, 12

= Corneromyces murrillii (Burt) Nakasone, comb. nov.

MycoBank MB812353

BASIDIOME resupinate, widely effused, <9 x 3.5 cm, loosely attached, <900 um

thick (Burt 1926), soft, fragile, membranous, abhymenial surface fibrous, felty,

brown (7E8), turning black in KOH then fading, sometimes hyphal strands

found under basidiome or in the substrate. HYMENIAL SURFACE smooth, even,

finely farinaceous, orange white to pale orange [5A(2-3)], greyish orange

[5B(4-5)], brownish orange [5C4-6)], yellowish brown (5D6), Cream-Buff, or

Chamois, black in KOH then fading to light brown; cracks occasional. CONTEXT

with a thin, dark brown layer next to substrate and a thicker, cream-colored

upper layer. MARGIN thinning out, fibrillose, loosely attached or detached,

concolorous with hymenium or developing a dark brown edge, sometimes

poorly developed hyphal strands present.

HYPHAL SYSTEM monomitic with clamped generative hyphae. SusicuLuM

<750 um thick, a non-agglutinated tissue composed of brown, rough-walled

subicular hyphae loosely arranged parallel to substrate, then hyphae turning

into hymenium, becoming hyaline; subicular hyphae of two types: (a) next

to substrate 3-5.5 um diam, clamped, moderately branched, walls yellow to

brown, <1.1 um thick, smooth or ornamented with tiny tubercles; (b) in upper

subiculum 3-6 um diam, clamped, moderately branched, walls hyaline, thin,

smooth. SUBHYMENIUM a non-agglutinated, scarcely differentiated tissue of

upright hyphae; subhymenial hyphae 2.2-5 um diam, clamped, moderately

branched, walls hyaline, thin, smooth. HyMENIuM a palisade of hyphidia and

basidia. Hypuip1a scarce, filamentous, obclavate, or narrowly clavate, (25-)

35-65 x (3-)4-6.5(-10) um, clamped atbase, walls hyaline, thin, smooth. BAsIDIA

collapsing soon after ejecting spores, obclavate at first then clavate to cylindric,

usually with a distinct stalk, (35-)40-90(-110) x 10-14 um, clamped at base,

walls hyaline, thin or slightly thickened at base, smooth, 4-sterigmate, sterigmata

Fics 12-13. Corticium murrillii (holotype BPI 0282179). 12. A. Basidiospores; B. basidia;

C. hyphidia. Dendrophysellum amurense (holotype TAAM 015561). 13. A. Basidiospores;

B. basidia; C. gloeocystidia; D. subicular hypha; E. skeletal-like hyphae; EF. dichohyphidia.

Unusual crust and pileate basidiomycetes ... 375

eS 12

376 ... Nakasone

<17 x 3.5 um. Basipiospores abundant, narrowly cylindric to allantoid,

(25-)28-40(-43) x (7.2-)8-11(-11.5) um, averages of three specimens

28.9-29.6 x 8.7-10.4 um, Q = 2.8-3.4, average of isotype at FH 34.4 + 4.2 x 9.9

+ 0.9 um, Q = 3.5 + 0.4, walls hyaline, occasionally pale yellow, thin to 1.3 um

thick, smooth, cyanophilous, dark blue-black in Melzer’s reagent.

HABITAT & DISTRIBUTION — On bark and wood of dead angiospermous

branches and logs in subtropical and tropical Central and South America.

TYPE SPECIMENS EXAMINED — MEXICO. Jalapa, 5000 ft, on bark of decaying log, 12-20

Dec 1909, WA & EL Murrill 182 (BPI 0282179, holotype; NY 00776553, NY 00562612,

and FH HUH-00290581, isotypes).

SPECIMENS EXAMINED — COLOMBIA. Magdalena, Sierra Nevada de Santa Marta,

Reserva Forestal San Lorenzo, on wood, 17-19 June 1978, L Ryvarden 16070 (O, as

“C. kinabalui”). ECUADOR. SucumBios PROVINCE: Reserva Natural de Cuyabeno,

300 m.a.s.l., on hanging branch, 28 June-15 July 1993, M Nunez 306 (O F-902593, as

“C. kinabalui” ).

DESCRIPTIONS & ILLUSTRATIONS — Burt (1926), Liberta (1969).

COMMENTS — Corneromyces murrillii is an uncommon and unusual species

characterized by a soft fragile basidiome, hyphidia, large basidia, and large

narrowly cylindric to allantoid basidiospores with amyloid and cyanophilous

walls. There was a wide range in basidium and basidiospore size among the

specimens examined. Corneromyces murrillii is similar to C. kinabalui Ginns

from Borneo, which has brown aculeate basidiomes, smooth brown subicular

hyphae, and brown basidiospores (Ginns 1976). Basidiospore sizes in the two

species are similar. The average basidiospore size of the isotype of C. kinabalui

at BPI, BPI 0290583, is 27 + 2.9 x 9.1 + 1.1 um, Q=3 + 0.3.

One isotype of C. murrillii (NY 00776553) is a large collection that is

intermixed with small basidiome fragments of Rhizochaete radicata (Henn.)

Gresl. et al. Hjortstam & Ryvarden (2001, 2008) erroneously reported

C. kinabalui from Columbia and Ecuador; two of the voucher specimens were

examined and identified as C. murrillii.

The affinities of Corneromyces Ginns are not known. Ginns (1976) considered

creating a new family for Corneromyces but placed it instead in Coniophoraceae

Ulbr. Citing the strongly amyloid, brown, thick-walled basidiospores, Julich

(1979) proposed the family Corneromycetaceae Jiilich. Recent molecular

phylogenetic studies have identified a new order, Amylocorticiales K.H. Larss.

et al. This order is characterized by varied basidiome habit and hymenophore

configuration, a monomitic nodose-septate hyphal system, and smooth thin-

or thick-walled basidiospores that are amyloid in most species (Binder et al.

2010). Corneromyces may belong in the Amylocorticiales, which is sister to the

Agaricales Underw., instead of with Coniophora DC. and allies in the Boletales

E.-J. Gilbert.

Unusual crust and pileate basidiomycetes ... 377

Dendrophysellum amurense Parmasto, Consp. syst. cortic.: 206. 1968.

FIGs 8-9, 13-15

= Vararia amurensis (Parmasto) Nakasone, comb. nov.

MycoBAnk MB812355

BASIDIOMES resupinate, effused, beginning as circular to irregular colonies,

then coalescing, <5 x 20 mm, adnate, thin, <150 um thick, subceraceous,

smooth to finely granulose over the irregular contours of the substrate, greyish

yellow [4(B-C)(3-4)], greyish orange [5B(3-4)], to brownish orange (5C3),

rimose, exposing white, felty to cottony context; cottony white mycelial tissue

developed in substrate and occasionally beneath basidiome. MARGIN adnate,

thinning out, white, farinaceous.

HYPHAL SYSTEM dimitic with clamped generative hyphae and aseptate

dichohyphidia. MycrELiaL Tissue cottony, white, composed of skeletal-like

hyphae; hyphae 2.2-3.5 um diam, occasionally clamped or appearing aseptate,

sparsely branched, straight, non-staining, walls hyaline, thin to thick, smooth.

SUBICULUM <100 um thick, basal layer next to substrate a compact, agglutinated

tissue of short-celled hyphae arranged more or less parallel to substrate, then

hyphae becoming upright, mostly indistinct and degraded although with

a few, intact, phloxine-stained hyphae; subicular hyphae from basal layer

3-5 um diam, clamped, frequently branched, walls hyaline, thin, smooth;

hyphae of upper subiculum 1.5-2.2 um diam, clamped, moderately branched,

walls hyaline, thin, smooth. SUBHYMENIUM AND HYMENIUM indistinct,

composed of dichohyphidia, gloeocystidia, and basidia intermixed with

abundant, coarse, hyaline crystals. DICHOHYPHIDIA scattered in hymenium,

not enclosed, dendriform, <20 x 20 um, with a main stalk, 2-2.5 um diam,

and multiple, short branches, walls hyaline, slightly thickened, smooth,

cyanophilous, dextrinoid. GLOEOCYSTIDIA scarce, inconspicuous, cylindrical

to narrowly clavate, occasionally papillate, 23-27 x 3-4 um, clamped at base,

protruding <15 um, containing oil-like material, negative in sulfovanillin, walls

hyaline, thin, smooth. Basrip1a suburniform at first, then narrowly cylindrical,

flexuous, about 26 x 5 um, clamped at base, protruding <10 um, 4-sterigmate,

walls hyaline, thin, smooth. BasipiosporREs numerous, agglutinated, often

collapsed, subfusiform to pip-shaped (in face view), 6-8(-9.3) x 3-3.7(-4)

um, x = 7.1 + 0.7 x 3.3 + 0.3 um, Q = 2.1 + 0.3, walls hyaline, thin, smooth,

acyanophilous, faintly amyloid.

HABITAT & DISTRIBUTION — On bark of dead Picea; known from the type

locality in northeastern Asia.

TYPE SPECIMEN EXAMINED — RUSSIA. KHABAROVSK REGION: Selikhino, Kabansopka,

on bark of fallen trunk of Picea jezoensis (Siebold & Zucc.) Carriere, 18 Aug 1961, E

Parmasto (TAAM 015561, holotype).

378 ... Nakasone

COMMENTS — Vararia amurensis is characterized by clamped generative

hyphae, dextrinoid and cyanophilous dendriform dichohyphidia,

gloeocystidia, suburniform basidia, and subfusiform amyloid basidiospores.

A typical catahymenium was not observed. Mature basidia are rare, and

gloeocystidia are inconspicuous and easily overlooked. It is morphologically

similar to V. mediospora var. makokouensis Boidin et al. from Gabon, which

has basidiospores of the same size and shape but lacks clamp connections

(Boidin et al. 1980). Parmasto (1968: 146) placed Dendrophysellum Parmasto

in Corticiaceae subfam. Aleurodiscoideae Parmasto, but it is morphologically

similar to taxa in Vararia sect. Fusamyspora Boidin & Lanq. that have dextrinoid

and cyanophilous dendrohyphidia, smooth or slightly ornamented amyloid

basidiospores, and sulfovanillin-negative gloeocystidia (Boidin & Lanquetin

1975). Thus, the monotypic genus Dendrophysellum is placed in synonymy

under Vararia and D. amurense is transferred to Vararia.

Hyphoradulum conspicuum Pouzar, Ceska Mykol. 41: 26. 1987. FIGS 16-17, 25

= Pseudolagarobasidium conspicuum (Pouzar) Nakasone, comb. nov.

MycoBank MB812356

BASIDIOME resupinate, widely effused, soft, fragile, cretaceous, spinose,

yellowish white (4A2), greyish yellow (4B3), or greyish orange (5B3), no color

change with KOH; cracks absent. HyMENIAL SURFACE composed of fragile,

brittle, soft, cretaceous or chalky aculei, <2 aculei per mm, <3.5 mm long, terete

to conical then gradually tapering to an acute or rounded apex, single at first

then fused at base or along entire length, occasionally reticulate, smooth or

studded with tiny, round tubercles, apices concolorous with base of aculei or

pale yellow, often broken off. MARGIN cream white, byssoid, fibrillose.

HyPHAL SYSTEM monomitic with clamped generative hyphae. ACULEI

composed of non-agglutinated tramal hyphae arranged in a fascicle with

embedded tramal cystidia curving into hymenium, at aculeal apex terminal

hyphae undifferentiated, smooth; tramal hyphae (1.5-)2-3(-4) um diam,

clamped, sparingly branched, even, walls distinct, hyaline, thin, smooth,

sometimes weakly cyanophilous. SusrcuLum 300-700 um thick, a non-

Fics 14-24. Dendrophysellum amurense (holotype TAAM 015561). 14. Dichohyphidium with

cyanophilous walls; 15. dichohyphidium with dextrinoid walls in Melzer’s reagent. Hyphoradulum

conspicuum (isotype PRM 834887). 16. Aculei; 17. close-up of aculei. Polyporus polyacanthophorus

(BPI 0261328). 18. Hyphal pegs from hymenial surface. Favolus curtipes (isotype HUH 00290582).

19. Close-up of poroid surface; 20. poroid surface of basidiome. Mycothele disciformis (isotype

BPI 0261300). 21. Close-up of basidiomes. Polyporus epitheloides (BPI 0261319). 22. Close-up

of apically encrusted hyphal pegs. Mycobonia winkleri (holotype F-15807). 23. Cross-section of

“hyphal peg” or bulbil; 24. close-up of hymenial surface with “hyphal pegs” or bulbils. Scale bars:

14, 15 = 10 um; 16, 18, 19 = 2 mm; 17, 21, 24 = 1 mm; 20 = 20 mm; 22 = 0.5 mm; 23 = 50 um.

Unusual crust and pileate basidiomycetes ... 379

380 ... Nakasone

agglutinated tissue; subicular hyphae 1-5 um diam, clamped, walls hyaline,

thin, smooth. SuBHYMENIUM <40 um thick, composed of short-celled

hyphae in a dense non-agglutinated tissue; subhymenial hyphae 2-3 um

diam, clamped, frequently branched, walls hyaline, thin, smooth. HYMENIUM

<35 um thick, a dense palisade of hyphidia, cystidia, and basidia. HypHip1a

scarce, inconspicuous, filamentous to subulate, 15-23 x 1.5-3 um, clamped

at base, walls hyaline, thin, smooth. Cystip1a of two types: (a) arising from

aculei trama, subiculum, and subhymenium, abundant, embedded, broadly

cylindrical, clavate, or obclavate, often strangulated or moniliform, sometimes

with a lateral lobe, stalked, apex obtuse, occasionally branched, (27—)40-80

x 5-10 um, tapering to 1.5-3 um diam at base, with a basal clamp, often

with honey yellow, refractive contents, walls hyaline, thin, smooth, negative

in sulfovanillin; (b) arising from hymenium, rare, clavate to cylindrical but

sometimes with a small, lateral beak, 11-15 x 4-5 um, with a basal clamp,

walls hyaline, thin, smooth. Basip1a clavate, often with a distinct stalk,

(16-)20-35 x 5-6 um, clamped at base, walls hyaline, thin, smooth, 4-sterigmate.

BASIDIOSPORES ellipsoid with a small apiculus, (4.8-)5-5.8(-6) x 3.5-4.4 um,

average of isotype 5.4+0.4 x 4+0.2 um, Q=1.3 + 0.1, filled with numerous

oil-like globules, walls hyaline, slightly thickened, smooth, cyanophilous, not

reacting in Melzer’s reagent.

HABITAT & DISTRIBUTION — Saprophytic on wood and bark of dead Cornus;

known from the type location, Bohemia.

TYPE SPECIMENS EXAMINED — CZECH REPUBLIC. Bouemica: “Velka hora” hill near

Karl&Stejn, on base of dead Cornus mas L., 2 Oct 1981, Z Pouzar (PRM 834886, holotype;

PRM 834887, isotype).

ComMENTS — Pseudolagarobasidium conspicuum is characterized by soft,

brittle, cretaceous or chalky aculei, strangulated or monilioid cystidia with

honey-yellow contents, and ellipsoid basidiospores with slightly thickened,

cyanophilous walls. Microbinding hyphae were not observed. Pouzar (1987)

noted that old collapsed basidiospores have dextrinoid walls, which was

not observed in this study. This is the first species of Pseudolagarobasidium

J.C. Jang & T. Chen reported from Europe. The description above is based

partially on Pouzar’s (1987) observations. Hyphoradulum conspicuum is

transferred to Pseudolagarobasidium, and Hyphoradulum, a monotypic genus,

becomes a synonym of Pseudolagarobasidium.

This species is most similar to P. pronum (Berk. & Broome) Nakasone

& D.L. Lindner with respect to color and texture of the basidiome.

Fics 25-26. Hyphoradulum conspicuum (isotype PRM 834887). 25. A. Basidiospores; B. basidia;

C. hymenial cystidia; D. tramal cystidia; E. section through hymenium with tramal cystidia and

immature basidia. Polyporus polyacanthophorus (TENN 57579). 26. A. Basidiospores; B. basidia.

382 ... Nakasone

Pseudolagarobasidium pronum develops microbinding hyphae and has slightly

smaller basidiospores (4-5.5 x 3-3.7(-4.3) um) with weakly cyanophilous

walls, whereas P. conspicuum lacks microbinding hyphae and has distinctly

cyanophilous basidiospore walls. In addition, cystidia in P pronum do not

contain refractive, honey-yellow contents. Pseudolagarobasidium conspicuum

is known from Europe, whereas P. pronum is reported from Asia, Australia,

Sierra Leone (Nakasone & Lindner 2012), and a first report from Jamaica

(Runaway Bay, on Calliandra sp., 12 Feb 2006, F Dammrich 8139 at CFMR).

See Nakasone & Lindner (2012) for a description of P pronum and other species

of Pseudolagarobasidium.

The genus Mycobonia

Mycobonia Pat. was erected with the generic type Hydnum flavum Berk.

(Patouillard 1894). Mycobonia was substituted for the illegitimate Bonia Pat.,

which had Bonia papyrina Pat. as its generic type (Patouillard 1892). These

two generic names are not homotypic; thus, Mycobonia was published as a new

genus rather than a replacement name. For a detailed nomenclatural history

of Mycobonia, see Jiilich (1976) and Martin (1939). One important feature of

Mycobonia is the dense stand of hyphal pegs penetrating the smooth hymenial

surface. Hyphal pegs are not restricted to Mycobonia but are produced in a

number of basidiomycete species. For a key to basidiomycete taxa with hyphal

pegs see Nakasone (2013). Mycobonia is readily distinguished from other

taxa with hyphal pegs and non-septate basidia by its substipitate and pileate

basidiomes.

The classification of Mycobonia is controversial because of its unique

combination of morphological features. Donk (1964: 294) placed Mycobonia in

the Stereaceae Pilat even as he noted a close relationship to Pseudofavolus Pat. in

the Polyporaceae Fr. ex Corda. Jiilich (1982: 186) created Mycoboniaceae Jiilich

for Mycobonia, placing the family in the Polyporales. Later, Singer (1986: 171)

reduced Mycoboniaceae to a synonym of Polyporaceae. Other researchers also

noticed striking morphological similarities between Mycobonia and Polyporus

(Corner 1984: 102; Kriiger 2002; Kriiger & Gargas 2010; Ryvarden 1991: 213,

2010: 119) and between Mycobonia and Pseudofavolus (Corner 1984: 36,

Kruger 2002, Kriiger & Gargas 2010, Singer 1986: 171). Authors have generally

agreed that Pseudofavolus is closely related to Polyporus s.s. (Nunez & Ryvarden

1995: 68, Ryvarden 1991: 213), while some consider Pseudofavolus a synonym

of Polyporus (Corner 1984: 35, Kriiger 2002, Kriiger & Gargas 2010).

Molecular phylogenetic studies show that Mycobonia and Pseudofavolus

are sister taxa embedded in the core polyporoid clade (Binder et al. 2013;

Kruger 2002; Kriiger & Gargas 2004, 2010; Sotome et al. 2008), confirming

Unusual crust and pileate basidiomycetes ... 383

morphological observations. Based on phylogenetic studies and morphological

similarities, Kriiger (2002) and Kriiger & Gargas (2010) reduced M. flava to

a subspecies of Polyporus curtipes (Berk. & M.A. Curtis) Ryvarden. While

agreeing with these authors that the molecular and morphological evidence

overwhelmingly indicates that Mycobonia is a synonym of Polyporus, significant

differences in ITS sequences and basidiospore shape and size argue for the

recognition of M. flava and M. brunneoleuca as distinct species. Transfer of

these taxa requires replacement names because both species epithets are

preoccupied in Polyporus.

Mycobonia brunneoleuca (Berk. & M.A. Curtis) Pat., Bull. Soc. Mycol. France 16:

181. 1901 (1900). FIGs 18, 26-27

= Hydnum brunneoleucum Berk. & M.A. Curtis, Trans. Linn. Soc. London

22: 129. 1857, non Polyporus brunneoleucus Berk. 1846.

= Polyporus polyacanthophorus Nakasone, nom. nov.

MycoBank MB812357

ErymMo.Loecy: from the Greek, poly- (many) + acantha (spine) + -phorus (bearing),

referring to the numerous hyphal pegs in the hymenial surface.

BASIDIOME pileate, sessile or substipitate, coriaceous, flabellate, reniform.

PitEus galeaeform or helmet-shaped, <10 x 8 cm; upper surface smooth,

becoming radially rugulose, bright yellow when fresh, drying to dull greyish

orange (5B6), brown (7D7), reddish brown (9E6), Vinaceous-Russet, Pecan

Brown, or Kaiser Brown. Stipe reduced, usually black, 3.5-9 mm diam,

attached to substrate by a thin, circular pad, <8-13 mm diam. CONTEXT

<4 mm thick at base, thinning to 0.3 mm at pileus edge, pale orange (5A3),

coriaceous. HYMENIAL SURFACE appearing smooth but densely covered with

minute, conical hyphal pegs, <330 x 40-80 um, 9-12 pegs per mm, becoming

fimbriate at apex, purplish tan when fresh, drying to greyish orange (5B3),

brownish orange to light brown [6(C-D)5], or Cinnamon Buff.

HYPHAL SYSTEM dimitic with clamped generative and aseptate skeleto-

binding hyphae. HypHaL PEGs a dense fascicle of non-agglutinated, sparsely

branched skeleto-binding hyphae, <330 um long, originating 100-150 um

below hymenium, projecting <180 tm, encrusted at apex with coarse, irregular,

hyaline crystals; hyphae rigid, tapering to acute apex, <3 um, walls hyaline to

light brown, slightly thick, smooth, cyanophilous, weakly dextrinoid. CONTEXT

a densely interwoven, non-agglutinated tissue composed mostly of skeleto-

binding hyphae and some generative subicular hyphae; subicular hyphae 3-6.5

um diam, clamped, strangulated, staining in phloxine, walls hyaline, thin,

smooth; skeleto-binding hyphae (0.5—)1.5-5.5 um diam, occasionally inflated

<12 um diam, then tapering to tips, with a narrow lumen, aseptate, rarely to

384 ... Nakasone

| Ke _

Oy ee. C

(DD 0 10 20 um

Fic. 27. Polyporus polyacanthophorus (NY 00536422). A. Basidiospores; B. thin-walled hyphidia;

C. cystidia; D. hyphidia from hyphal pegs; E. skeleto-binding hyphae from context of NY 00543220.

extensively branched, walls hyaline, <1.5 um thick, smooth, acyanophilous,

not reacting in Melzer’s reagent. SUBHYMENIUM a dense, thickening, non-

agglutinated tissue of upright, short-celled hyphae; subhymenial hyphae

3.5-4.5um, clamped, frequentlybranched, wallshyaline, thin,smooth. HyYMENIUM

a dense palisade of hyphidia, cystidia, and basidia. HypHip1A or two types:

(a) inconspicuous, filamentous, often with short branches at apex, (23-)32-60

Unusual crust and pileate basidiomycetes ... 385

(-80) x 2-4 um, clamped at base, walls hyaline, thin, smooth; (b) numerous,

acicular or narrowly cylindrical then tapering to a subacute or acute tips, 1.5-4

um diam, with a narrow lumen, aseptate, sparsely branched, walls hyaline,

slightly thick to thick, smooth, acyanophilous, not reacting in Melzer’s reagent

(may be interpreted as terminal ends of skeleto-binding hyphae). Cystip1a

of two types: (a) rare, inconspicuous, embedded, subfusiform to cylindrical,

tapering slightly to apex, 45-55 x 6-7.5 um, clamped at base, walls hyaline, thin,

smooth; (b) rare, obclavate, an enlarged bulbous base tapering to apex, 25-45

x (6-)10-15 um, aseptate, walls hyaline, <1 um thick, smooth, acyanophilous,

not reacting in Melzer’s reagent. Basrp1A scarce, clavate with a stalk, <80 x

10-16 um, clamped at base, walls hyaline, thin, smooth, with 4-sterigmata.

BASIDIOSPORES cylindrical to narrowly ellipsoid or broadly subfusiform with

a small, distinct, hyaline apiculus, (13-)16-24(-26.5) x (7.2-)8-11(-12.3) um,

average of three specimens 17.3-21 x 9.4-10.1 um, Q = 1.9-2.2, containing

oil-like material, walls hyaline to light brown, thin to 0.7 um thick, smooth,

acyanophilous or weakly cyanophilous, not reacting in Melzer’s reagent.

HABITAT & DISTRIBUTION — On logs, branches and twigs of dead hardwoods

throughout Central and South America and the Caribbean region.

TYPE SPECIMEN EXAMINED — VENEZUELA. Fendler 129 (FH HUH-00290586, isotype

of H. brunneoleucum).

REPRESENTATIVE SPECIMENS EXAMINED — BRAZIL. Prov. SAO PAuLo: S. Framisco dos

Campos, Dec 1896, F Noark 333 (S F-177421); Caldas, pnu. Miras Gerais, Regnell (S

F-249624). COLUMBIA. Dept. DEL Cauca: Cordillera Central, Hoya del Rio Palo, Ta

Tolda, 1944, J Ceratrecas 19440 (BPI 0261323); Munchique, 2700 m elev., on dead wood,

5 May 1968, AL Welden 7318 (NY 00536422). COSTA RICA. PUNTARENAS PROVINCE:

Monteverde, on recently cut small trees, 8 Jan 1973, AL Welden 3303 (NY 00543220);

Sta. Elena, road to Elena Biological Reserve, on fence post, 17 Mar 1999, RH Petersen

10256 (TENN 57579). SAN JOSE PROVINCE: San Isidro de Coronado, on fallen log, 21 Jun

1972, AL Welden 8338 (NY 00536420); Monté Zurqui, alt. 2000-2500 m., on log, 13 Feb

1926, PC Standley & J Valerio no. 48084 (BPI 261325). Dota, Guadeloupe, Finca Jaular,

km 66 on Interamerican Hwy, 1 Jul 1998, RH Petersen 9471 (TENN 56445). HEREDIA

PROVINCE: Cerro Central de Zurqui, 1600 m., on dead log, 27 Dec 1929, CW Dodge no.

633 (BPI 261320). HONDURAS. Tegucigalpa, Escuela Agricola Panamericana, 31 Dec

1951, AS Mueller (BPI 261322). PANAMA. Cuiriquf PRovINCE: valley of upper Rio

Chiriqui Viejo, 1600-1800 m., 10 Jul 1935, GW Martin 2714 (BPI 261324, as “M. flava”);

6 Jul 1953, GW Martin 2519 (BPI 261238, as “M. flava”); 7 Jul 1935, GW Martin 2631

(NY 00536417, as “M. flava”). PARAGUAY. Asuncion, San Antonii, July 1893, Malme

(S F-249621). PUERTO RICO. Guajataca Community Forest, Verada, Nueva Trail, on

deciduous wood, 26 Jun 1996, L Ryvarden (CFMR PR 5160).

DESCRIPTIONS & ILLUSTRATIONS — Gerlach & Loguercio-Leite (2011), Martin (1939:

248, figs 13-16, as M. flava), Jiilich (1976, figs 1-2 only), Reid (1976: 191, fig. 2).

ComMENTsS — Polyporus polyacanthophorus is a pileate species characterized

by a dense stand of hyphal pegs in the hymenium and large, cylindrical to

386 ... Nakasone

ellipsoid basidiospores. Although considered a synonym of P. epitheloides (as

M. flava) by some mycologists, it can be differentiated by its larger basidia and

broader basidiospores. In addition, the thin-walled, fusiform to cylindrical

cystidia observed in P polyacanthophorus are not present in P. epitheloides.

Reid (1976) postulated that P. polyacanthophorus (as M. brunneoleuca) prefers

higher elevation habitats than P epitheloides (as M. flava).

The description above is based on Martin (1939), Reid (1976), and

personal observations. A replacement name is proposed for H. brunneoleucum

because the species epithet is occupied in Polyporus. Although Martin (1939:

247) considered M. brunneoleuca a synonym of M. flava, the specimens he

referenced from Panama are redetermined here as P. polyacanthophorus.

Similarly, the drawings of M. flava in Julich (1976, Fig. 2) of an Argentinian

specimen probably represent P. polyacanthophorus based on the basidiospore

shape and size.

Mycobonia disciformis G. Cunn., Trans. Roy. Soc. New Zealand 83: 635. 1956.

FIGS 21, 28

= Mycothele disciformis (G. Cunn.) Jiilich, Persoonia 8: 452. 1976.

BASIDIOMES resupinate, scattered to gregarious, disciform, centrally attached

to substrate, orbicular to elongate, 1-10 mm diam, <0.5 mm thick, firm,

membranous to ceraceous, spinose because of numerous hyphal pegs, exterior

surface dark brown. HYMENIAL SURFACE smooth with numerous hyphal pegs

penetrating surface, cream, dull sulfur-yellow, or light brown. HyPHAL PEGS

cylindrical, tapering to a subacute apex, 20-30 per mm. Marcein distinct,

abrupt, free, slightly involute.

HYPHAL SYSTEM monomitic with clamped generative hyphae. HyPHAL PEGS

originating deep in subicular trama, a dense, agglutinated fascicle of hyphae

with an central column of coarse, hyaline crystals, <1000 x 45 um, protruding

<135 um. SuBicuLUM <800 um thick with a basal layer of compact hyphae

arranged parallel to substrate, then hyphae becoming upright, forming a

dense but non-agglutinated tissue; subicular hyphae 2-3 um diam, clamped,

moderately branched, walls hyaline to brown, thin to slightly thick, smooth.

HyMENIuM <70 um thick, a dense palisade of hyphidia and basidia. HypHip1a

filamentous, occasionally branched at apex, 20-40 x 2-2.5 um, clamped at base,

walls hyaline, thin, smooth. Basrp1a fragile, scarce, cylindrical to narrowly

clavate, sometimes with a short, stalk-like base, (38-)45-60 x (7—)9-11(-14)

um, clamped at base, walls hyaline, thin, smooth, 4-sterigmate, sterigmata 7-10

x 1.5-2 um. BASIDIOSPORES scarce, globose to subglobose or broadly ellipsoid,

8-13.5 x (6.5-)7-9(-10) um, filled with oil-like globules, walls hyaline, thin,

smooth, acyanophilous or weakly cyanophilous, not reacting in Melzer’s

reagent.

Unusual crust and pileate basidiomycetes ... 387

C B

any:

0 10 20 um

Aree eee ee eal

Fic. 28. Mycothele disciformis (isotype BPI 0261300). A. Basidiospores; B. basidia; c. hyphidia.

HABITAT AND DISTRIBUTION — On bark of living trunks of Rhopalostylis

sapida H. Wendl. & Drude in New Zealand.

TYPE SPECIMEN EXAMINED — NEW ZEALAND. AucxkLanp: Henderson Valley,

130 m., Sharps Bush, on bark of R. sapida, 5 May 1952, SD Baker PDD 11491 (BPI

261300, isotype). In the original description, the month is mistakenly reported as April

(Cunningham 1956: 636).

SPECIMENS EXAMINED — NEW ZEALAND. AUCKLAND: Titirangi, Atkinson Park, on

R. sapida, 27 Jun 1953, JM Dingley (PDD 12643); Coromandel Peninsula, Camel’s Back,

on R. sapida, Oct 1954, JM Dingley (PDD 14304); Waitakeres, Cascades, on R. sapida, 3

Apr 1954, SD Baker (PDD 14307).

DESCRIPTIONS AND ILLUSTRATIONS — Cunningham (1956, 1963: 173), Julich (1976).

388 ... Nakasone

Comments — Mycothele disciformis is characterized by small fragile disciform

basidiomes with prominent hyphal pegs. Epithele ryvardenii Nakasone is an

effused species that is reminiscent of M. disciformis because it produces brown-

pigmented hyphae and has similarly sized and shaped basidia and basidiospores

(Nakasone 2013). However, M. disciformis is found on bark of living nikau, an

endemic New Zealand palm, whereas E. ryvardenii is reported from Venezuela

on wood. Mycothele disciformis is undoubtedly closely related to Epithele (Pat.)

Pat. Whether it should be transferred to Epithele or retained as a monotypic

genus may be resolved in the future with molecular evidence.

Mycobonia flava (Sw. : Fr.) Pat., Bull. Soc. Mycol. France 10: 77. 1894. Fics 22, 29-30

= Peziza flava Sw. : Fr., Prod.: 150. 1788, non Polyporus flavus Jungh. 1838.

= Hydnum flavum (Sw. : Fr.) Berk., Ann. Mag. Nat. Hist., 10: 380. 1843 [“1842”].

= Bonia flava (Sw. : Fr.) Henn., Hedwigia 36: 192. 1897, as “(Berk.) Pat.”

= Auricularia flava (Sw. : Fr.) Farl., Bibl. Index N. Amer. Fung.: 307. 1905.

= Grandinioides flava (Sw. : Fr.) Banker, Mem. Torrey Bot. Club 12: 179. 1906.

= Polyporus curtipes subsp. flavus (Sw. : Fr.) D. Kriiger, Cryptog. Mycol. 31: 399. 2010.

= Polyporus epitheloides Nakasone, nom. nov.

MycoBank MB812358

EryMo_oey: refers to similarity with the genus Epithele which is characterized by

hyphal pegs.

BASIDIOME pileate, dimidiate, flabelliform or reniform, sessile or substipitate,

coriaceous. PILEus applanate, subdiscoid, or shallowly cucullate, <8 x 4 cm;

upper surface smooth then becoming radially rugulose, when dried dull, pallid

ochraceous, greyish orange (5B5) to brown [6D(7-8)] or purplish chestnut,

darkening at edges to brown (6E8). Stipe short, <5 mm long by 3 mm diam,

often black, attached to substrate by a thin, circular, mycelial pad, <8 mm diam.

ConTEXT 1-6 mm thick at base, thinning to 0.5 mm at pileus edge, light orange

(5A4), thin, coriaceous. HYMENIAL SURFACE appearing smooth but densely

covered with cylindrical hyphal pegs, <400 x 40-60 um, 5-11 pegs per mm,

pale ochraceous when fresh, drying to greyish orange (5B4), Light Ochraceous

Buff, purplish buff, or light rusty buff.

HYPHAL SYSTEM dimitic with clamped generative and aseptate skeleto-

binding hyphae. HyPHAL psGs a dense fascicle of non-agglutinated, sparsely

branched, skeleto-binding hyphae, originating in context, projecting <200

um, heavily encrusted with coarse, irregular, hyaline crystals; hyphae 1-3

um diam, aseptate, rarely branched, walls hyaline, slightly thick to thick,

smooth, cyanophilous, weakly dextrinoid. CONTEXT a densely interwoven,

Fic. 29. Polyporus epitheloides (NY 00536421). A. Basidiospores; B. papillate hyphidia;

C. thin-walled hyphidia; D. thick-walled hyphidia or skeleto-binding hyphae terminating in

hymenium; E. cystidium.

390 ... Nakasone

non-agglutinated tissue composed primarily of skeleto-binding hyphae and

indistinct generative hyphae, in pileus cuticle hyphae similar but compacted

into a cutis; generative hyphae 2.2-4 um diam, clamped, walls hyaline, thin,

smooth; skeleto-binding hyphae 2.2-5 um diam, occasionally inflated <20 um

diam, aseptate, rarely to extensively branched, branches often elongate and

whip-like, walls hyaline, thick, smooth, acyanophilous, not reacting in Melzer’s

reagent. HyMENIUM a dense palisade of hyphidia, cystidia, and basidia.

Hypuip1a of two types: (a) inconspicuous, scarce, filamentous, often with a

few short branches at apex or papillate, 27-52 x 2-4 um, clamped at base, walls

hyaline, thin, smooth; (b) numerous, acicular or narrowly cylindrical then

tapering to a subacute to acute apex, 30-60 x 2-4 um, aseptate, walls hyaline,

slightly thick, smooth, acyanophilous, not reacting in Melzer’s reagent (may be

interpreted as terminal branches of skeleto-binding hyphae). CysTip1A rare,

obclavate, an enlarged bulbous base then tapering to apex, 25-45 x (6-)10-15

um, aseptate, walls hyaline, <1 um thick, smooth, acyanophilous, not reacting

in Melzer’s reagent. Bastp1a clavate, 26-40 x 6-12 um, clamped at base,

walls hyaline, thin, smooth, with 4-sterigmata. BAsip1osPorEs cylindrical to

subfusiform, 14-23(-26) x 5-8(-8.7) um, average of three specimens 15.6-22.5

x 5.2-7.4 um, Q = 3, filled with oil-like material, walls hyaline, thin, smooth,

acyanophilous or weakly cyanophilous, not reacting in Melzer’s reagent.

HABITAT & DISTRIBUTION — On logs, branches and twigs of deadangiosperms

from southeastern United States (Florida, Louisiana) to southeastern Brazil.

REPRESENTATIVE SPECIMENS EXAMINED — ARGENTINA. MIsIONES PROVINCE:

Urugua-i Provincial Park, Wanda, 26 May 2001, D Kriiger 11279 (TENN 59088).

BRAZIL. Rio GRANDE DU SOL PROVINCE: Sao Leopoldo, 1904, Rick (S F-249620); Rick-

Fungi Austro-Americano Exsiccati nr. 141, 1906 (S F-177427, F-177429); Ijuhy, Exped.

Imae Regnelliana no. 345B, 7/4 1893, GA Malme (S F-177430). COLOMBIA. Cauca

District: ad pag. El Tambo, 1700 m, 13 Nov 1937, Kv Snidern (S F-177424). Valle

del Cauca Dept., elev. 1980 m., 23 km from Cali, on Cali-Cisneros, on branch, 30 Aug

1976, KP Dumont & LA Molina, Dumont-Co 7669 (NY). COSTA RICA. PUNTRENAS

PROVINCE: Coto Brus County, San Vito, Hacienda La Amistad, 3 Jul 1998, RH Petersen

9486 (TENN 56514); Tablazo, 1800 m, 8 Aug 1932, M Valerio (S F-177432). CARTAGO

PROVINCE: mountains east of Tulis and north of Plantanillo, 2 Sep 1968, DE Stone 7141

(NY 00536421). CUBA. Fungi Cubensis Wrightiani no. 343, [Wright] (S F-15535);

Monte Verde, Fungi Cubensis Wrightiani no. 237, 7-4 1915, C Wright (S F-15533).

PINAR DEL RIO PROVINCE: San Diego de Los Bajos, on old log, 31 Aug to 3 Sep 1910,

NL Britton, FS Earle & OS Gages, Britton 6823 (NY). JAMAICA. St. ANN ParIsH: near

Reynolds bauxite mines, Lynford P.O., 1200-1400 ft., on dead branch of small tree, 23

Sep 1954, GR Proctor, F1288 (NY 00536418); Moneague to Hollymount via Union Hill,

700-750 m. elev., 6 Aug 1957, AL Welden 558 (NY 00536419). PARAGUAY. ALTro

PARANA STATE: Ciudad del Este Hernandarias, Tati Jupi Reserve on Iaipu Lake, on

Fics 30-31. Polyporus epitheloides. 30. Basidiospores: A. from TENN 56514; B. from TENN

59088. Mycobonia winkleri (holotype F-15807). 31. A. Basidiospores; B. broom-like structures in

pileus context; C. hyphal segments from pileus context.

392 ... Nakasone

hardwood branch, 25 May 2001, RH Petersen (TENN 58933); SAN ANTONIO PROVINCE:

Asuncion, Exped. Imae Regnellian Fungi, July 1893, GA Malme (S F-177425). UNITED

STATES. Fioripa: Florida Caverns State Park, on dead wood, 5 May 1962, AL Welden

4428 (NY 00536416); Miami, on twigs, 15 Jan 1919, WH Long (CFMR FP 30869); Dade

County, Matheson’s Hammock, on twigs, 23 Feb 1922, JA Stevenson 1630 (BPI 261319).

Loutsi1ANa: Plaquemines Parish, Tulane University, F Edward Herbert Center, on dead

wood, 25 Jul 1972, AL Welden 7764 (NY 00536411); St. Martinville, on fallen branches,

20 Aug 1898, AB Langlois 2817 (S F-249626).

DESCRIPTIONS & ILLUSTRATIONS — Burt (1919: 262), Corner (1984: 104), Julich (1976:

450, description only), Patouillard (1894, plate III, fig. 2), Reid (1976: 190, figs 1, 5),

Ryvarden (2010: 119).

ComMENTsS — Polyporus epitheloides is a pileate species characterized by a

dense layer of hyphal pegs in the hymenium and large cylindric to subfusiform

basidiospores. The peculiar thick-walled cystidia seen in this species were

also observed in P. polyacanthophorus and could be interpreted as terminally

differentiated skeleto-binding hyphae. Jilich (1976: 451) refers to these

structures as tramal cystidia and includes an illustration. The description above

is based on Jiilich (1976), Reid (1976), and personal observations. Transfer

of the species to Polyporus requires a new name because the epithet ‘flavus’ is

preoccupied.

Polyporus epitheloides is closely related to P. polyacanthophorus

[= M. brunneoleuca] but can be differentiated by its narrowly cylindrical

to subfusiform basidiospores that are usually <8 um diam, Q = 3. Polyporus

epitheloides has a wider distribution than P polyacanthophorus, extending

into southeastern United States. ITS sequences of P. epitheloides (AY513571 as

M. flava) and P. polyacanthophorus (AY513569, AY513570 also as M. flava)

differ by 5%, confirming their status as distinct species. Another similar

species is Pseudofavolus cucullatus (Mont.) Pat. [= Polyporus curtipes]. It can

be distinguished from P. epitheloides by its angular to hexagonal pores, 1-3 per

mm, and smaller, cylindrical basidiospores (11.5-)13-16 x 4-6 um (Ryvarden

& Johansen 1980: 514). The isotype of Favolus curtipes Berk. & M.A. Curtis

(South Carolina, Santee, Ravenel 378, HUH 00290582) was examined (see

Fics. 19-20 for photographs of the poroid hymenophore).

Kriiger (2002) and Kriiger & Gargas (2010) considered M. flava to be a

subspecies of P. curtipes despite a 4—7% difference in ITS sequences. They noted

that M. flava, with spines, and Ps. cucullatus, with pores, were otherwise similar

in basidiome coloration, hyphal construction, and shape and size of basidia and

basidiospores. Because they are sister taxa embedded in Polyporus sensu stricto

as demonstrated by phylogenetic analyses of the large subunit ribosomal RNA

gene and ITS region, Kriiger (2002) and Kriiger & Gargas (2010) concluded

that M. flava and Ps. cucullatus belong in Polyporus. Following Ryvarden (1991:

Unusual crust and pileate basidiomycetes ... 393

213), they accepted the name P. curtipes for Ps. cucullatus and then reduced

M. flava to a subspecies of P. curtipes.

It should be noted that Kriiger (2002) and Kriiger & Gargas (2010)

misidentified TENN 057579, FB 10256, as M. flava. Based on basidiospore

size, this specimen represents P. polyacanthophorus [= M. brunneoleuca)]. Thus,

ITS (AY513569, AY513570) and LSU sequences (AJ487934) correspond to

P. polyacanthophorus whereas AY513571, ITS sequence of TENN 059088,

FB 11279, is correctly identified as P. epitheloides [= M. flava].

The following descriptions and illustrations should be used with caution, for

the authors did not differentiate between M. flava and M. brunneoleuca or did

not provide basidiospore measurements, so it is not possible determine the

species involved — Banker (1906), Berkeley (1843, plate 10, fig. 8), Gibertoni et

al. (2006), Hennings (1900), Ibanez (1999), and Lloyd (1915). Although Martin

(1939: 247) considered M. brunneoleuca a synonym of M. flava, the specimens

he cited from Panama are redetermined here as P. polyacanthophorus.

Mycobonia winkleri Bres., Ann. Mycol. 9: 551. 1911. FiGs.23—24, 34

BASIDIOME dimidiate-sessile to subresupinate, 17 x 7 mm, 320-400 um

thick, soft, fragile, greyish orange (5B4). PILEUS SURFACE warty to wrinkled;

HYMENIAL SURFACE smooth with warts, 2-3 per mm, with a dark cavity or

reddish brown, mucilaginous material at apex; context cottony, orange white

(5A2).

HYPHAL SYSTEM dimitic with clamped generative and aseptate skeletal

hyphae. Warts consisting of bulbils embedded in context and hymenium,

composed of pseudoparenchymatous tissue enclosing a mass of dark reddish

brown, amorphous material. PILEUS CORTEX a densely agglutinated tissue

composed of irregularly thick-walled hyphae, thick-walled, broom-like

structures, and scattered dichophyses; pileal hyphae 3-8 um diam, clamped,

with numerous short branches, irregular, walls hyaline, thick, smooth; broom-

like structures 17-30 x 3-8 um, with a thick, robust main trunk that gives rise to

branches short and stubby or longer, <12 x 2 um, with knobby protuberances,

walls hyaline, thick, smooth, cyanophilous, not reacting in Melzer’s reagent;

dichophyses with short, slender branches radiating from a central point, walls

hyaline, thick, smooth, cyanophilous, dextrinoid. Beneath cortex a narrow,

moderately dense zone of generative subicular and skeletal hyphae, then a zone

of open, loosely intertwined subicular hyphae and scattered dichophyses, about

140 um thick; subicular hyphae 3-5 um diam, clamped, moderately branched,

walls hyaline, thin to 2 um thick, sometimes irregularly thickened, smooth,

acyanophilous, not reacting in Melzer’s reagent; skeletal hyphae 1.3-2.2 um

394 ... Nakasone

diam, aseptate, sparsely branched, walls hyaline, thick, smooth, cyanophilous,

dextrinoid in Melzer’s reagent. SUBHYMENIUM a dense, agglutinate tissue of

hyphae similar to subicular hyphae. HYMENIUM a dense, agglutinated palisade

of basidia. Basrp1a clavate, 25-28 x 12 um, bisterigmate (Bresadola, 1911).

BASIDIOSPORES scarce, globose, subglobose or broadly lacrymoid, 6-7.5(-7.9)

x (5.5-)6-7.2 um, walls hyaline, thin, smooth, cyanophilous, not reacting in

Melzer’s reagent.

TYPE SPECIMEN EXAMINED — S.O. BORNEO, ad Calamus rotang, 24 Jun 1902, H

Winkler 2605 (S F-15807, holotype).

ComMENts — Mycobonia winkleri is a conundrum. The fungus displays a

bewildering array of morphological features and the name cannot be placed in

any extant genus. In the first place, the unusual structures that resemble hyphal

pegs in M. winkleri (and probably the reason Bresadola placed the species in

Mycobonia) are not composed of hyphae but consist of an amorphous reddish

brown material. Second, the cyanophilous dextrinoid dichohyphidia developed

in M. winkleri resemble those found in some species of Vararia P. Karst., such

as V. minidichophysa Boidin & Lanq., V. microphysa Boidin & Langq., and

V. tropica A.L. Welden. Boidin et al. (1980) described these structures as capillary

dichophyses. Two species of Amyloflagellula Singer, A. verrucosa Agerer &

Boidin and A. inflata Agerer & Boidin, also develop similar dichohyphidia

(Agerer & Boidin 1981). Third, the broom-like structures found in the pileus

cortex recall those produced in the pileipellis and gills of some Marasmius Fr.

species. However, the rather large, dimidiate-sessile basidiome of M. winkleri

is unlike that of Vararia (effused), Amyloflagellula (cup-shaped), or Marasmius

(mushroom-shaped).

It is premature to erect a new genus for M. winkleri at this time. The type

is fragile and somewhat deteriorated. Additional collections are needed to

confirm that the disparate characters are produced by this taxon. The hyphal

peg-like structures are particularly mysterious and in need of further study.

DNA sequence data would be desirable to discover its closest relatives.

Acknowledgments

Curators and assistants of the following herbaria are thanked for arranging specimen

loans: BPI, HUH, LY, NY, O, PDD, PRM, TAAM, TENN and S. Comments and

corrections of Drs. Andrew M. Minnis, James H. Ginns, and Harold H. Burdsall, Jr.,

who critically reviewed this manuscript, are greatly appreciated.

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MY COTAXON

http://dx.doi.org/10.5248/130.399

Volume 130, pp. 399-419 April-June 2015

A new Graphilbum species

from western hemlock (Tsuga heterophylla) in Canada

JAMES REID & GEORG HAUSNER*

Department of Microbiology, University of Manitoba, Winnipeg, MB, R3T 2N2

* CORRESPONDENCE TO: hausnerg@cc.umanitoba.ca

ABSTRACT — A new species of Graphilbum, G. tsugae isolated from Tsuga heterophylla, is

described from British Columbia, Canada. Although rDNA ITS data analysis demonstrates

its close relationship to Graphilbum rectangulisporium, morphologically the unique apical

projections arising from the perithecial necks of G. tsugae easily differentiate it from the

former and all other described Graphilbum and Ophiostoma species. We also correct

the epithet orthography and the parenthetical basionym author citation of the name

G. rectangulisporium.

KEY worps — ophiostomatoid fungi, bark beetles, morphology, taxonomy, ITS rDNA region.

Introduction

During a reassessment of isolates of ophiostomatoid fungi in our culture

collection whose taxonomic placements had never been resolved, we examined

three strains from a single location in southern British Columbia isolated from

two felled logs of Tsuga heterophylla. These logs had shown clear evidence of

having been attacked by bark beetles — many bore holes were evident and

peeling back the bark around the holes revealed well-developed galleries — but

neither beetle adults nor larvae were present and the surrounding sapwood was

only slightly stained. However, hand lens examination did show both remnant

hyphae and possible reproductive structures scattered throughout gallery

interiors.

Slabs of bark-covered wood about 12 inches long, 6-8 inches wide, and

1-3 inches thick were harvested from such areas using a hatchet and a chisel.

The bark was then removed to expose the outer sapwood surfaces and the

inner bark surfaces of these slabs. After the wood slabs and bark pieces were

examined carefully to ensure no adult beetles or larvae were present in any of

the exposed galleries, these materials were placed and sealed in collection bags

for return to the laboratory.

400 ... Reid & Hausner

When we began our detailed study of this species, the Ophiostomatales

accommodated three genera: Ceratocystiopsis H.P. Upadhyay & W.B. Kendr.;

Grosmannia Goid.; and Ophiostoma Syd. & P. Syd., among which species

economically important tree pathogens and blue-stain fungi are found

(Olchowecki & Reid 1974, Wingfield et al. 1993, Hausner et al. 2005).

Based on partial LSU rDNA and 6-tubulin sequences, Zipfel et al. (2006)

provided some support for the circumscription of the above three genera. But

Zipfel et al. (2006, p. 95) and other authors (Hafez et al. 2012, 2013) also noted

that the use of molecular criteria might require the erection of new genera to

accommodate either previously misplaced or subsequently newly discovered

species of Ophiostoma sensu lato.

Recently de Beer & Wingfield (2013), using partial LSU and SSU rDNA

sequences, have begun redefining the Ophiostomatales. They include only one

family, the Ophiostomataceae, in which they recognize six distinct lineages

represented by the genera Ceratocystiopsis H.P. Upadhyay & W.B. Kendr. emend.

Zipfel et al. (2006), Fragosphaeria Shear, Leptographium Lagerb. & Melin sensu

lato (includes Grosmannia), Ophiostoma sensu lato (includes Pesotum and

Sporothrix), Raffaelea Arx & Hennebert emend. T.C. Harr. in Harrington et al.

(2008), and Graphilbum H.P. Upadhyay & W.B. Kendr. emend. Z.W. de Beer

et al. (2013a). They also clearly implied the possible need for further generic

inclusions. Relevant comments are also found in related accompanying papers

(de Beer et al. 2013a & b).

The genera of the Ophiostomataceae include economically important tree

pathogens and blue-stain fungi (e.g., Lagerberg et al. 1927, Olchowecki & Reid

1974, Wingfield et al. 1993, Hausner et al. 2005) whose members lack forcible

ascospore discharge, have deliquescent asci, develop sticky ascospore droplets

at the apex of their perithecial necks, and typically produce slimy/sticky conidia

on various simple or complex conidiophore structures. Species assigned to

Ceratocystiopsis resemble those of Ophiostoma except in Ceratocystiopsis the

small dark perithecia only develop short perithecial necks and produce falcate,

sheathed ascospores (Upadhyay 1981, Zipfel et al. 2006, Plattner et al. 2009).

These species also have a lower tolerance to cycloheximide than do Ophiostoma

species (Harrington 1981, Hausner et al. 1993). Species formerly assigned to

Grosmannia (above) were distinguished by the fact they have Leptographium

states.

As we could not definitively establish the identity of our three isolates using

only morphological criteria, we compared nuclear small subunit rDNA (rSSU)

and rDNA internal transcribed spacer (ITS) region (ITS1, 5.8S gene, and ITS2)

sequences from our isolates with those of other Ophiostoma species. We also

assessed the growth rates and cultural characteristics of our isolates at different

Graphilbum tsugae sp. nov. (Canada) ... 401

temperatures and compared our results with data published for other species.

We did not assess mating type features, as we had earlier when describing

another ophiostomatoid species (Reid et al. 2010), as mating type studies had

not been conducted for any of the species with which we compared our current

isolates.

Based on the revisions discussed above, we believe our results show that our

isolates represent a new taxon within the genus Graphilbum that we describe

here as Graphilbum tsugae.

Materials & methods

Isolation of strains studied, culturing procedures, and

growth-temperature experiment

Laboratory isolations were made from field collected wood and bark samples and

grown on agar plates in a 20°C incubator; these were examined periodically for evidence

of fungi of interest.

We obtained three isolates that, based solely on morphological criteria, appeared

to represent an undescribed Ophiostoma species. Strains examined and/or sequenced

are detailed in TaBLeE 1. Living cultures and dried specimens of our new species have

been deposited in the University of Alberta Microfungus Collection and Herbarium

(UAMH), Edmonton, Canada; the additional DNA sequences retrieved from GenBank

employed in the analyses are also listed in TABLE 1.

Based on the intent of the experiment, strains were grown and characterized

employing either malt extract agar supplemented solely with yeast extract (Reid et

al. 2010) or such agar that also contained wood chips (Reid & Hausner 2010); fertile

perithecia were regularly produced on the latter plates. These were then dried, and

one was selected to be the type specimen. The parent culture was isolate WIN(M)

1391, obtained from a perithecial spore drop produced at the apex of a perithecium

found in the gallery of an unidentified bark beetle. Preparations for microscopy were

mounted in 85% lactic acid and at least 50 measurements were made for each significant

morphological feature. Colour designations are based on Rayner (1970).

In a previous study (Reid et al. 2010) where we had found a group of very similar

isolates growing on pieces of substrate in very close proximity to each other — as was

the case here — we had processed three of them in a temperature growth-rate response

experiment, and they produced very similar growth-temperature files. They and the

other isolates in the group were described as Ceratocystis norvegica J. Reid & Hausner

[sic; = Georg Hausner].

Because of that experience, the precise identity of the gene regions sequenced in

our current three isolates, and the lack of any significant morphological variation noted

between them, we used only the designated holotype isolate, VAMH 11701 [WIN(M)

1391] for determining the temperature growth-rate response of G. tsugae.

We employed the protocols of van Wyk et al. (2006), as modified by Reid et al.

(2010). However, because of the cultures’ relatively slow growth, the first measurements

were not made until the second day after plate inoculation. We used eight replicate

plates at each test temperature. Although initially, the total temperature-growth test

402 ... Reid & Hausner

TABLE 1. The rDNA ITS region sequences evaluated

TAXON STRAINS/ISOLATES/SOURCE! GENBANK

Ceratocystiopsis minuta °CBS 145.59 DQ128173

C. cf. minuta SL-K 70 DQ128175

Grosmannia penicillata RJ-T 0125 AM943882

Ophiostoma abietinum CMW 397 DQ396788

O. dentifundus CMW 13016? AY495434

CMW 13017? AY495435

O. ips CMW 22843 DQ539549

O. grandicarpum Strain D AJ293884

O. microcarpum YCC 459 AB506676

YCC 612 GU134170

O. piceae Ci-37 EF506934

O. pulvinisporum CMW 9026 AY546715

O. rectangulisporium TEM:FPH 7756 AB242825

YCC 617 GU134171

O. cf. rectangulisporium CMW 26261 > EU785451

CMW 26259° EU785450

CMW 26258 > EU785449

C2477 GU129987

C2300 GU393357

253GRJ © JX444645

1313RJP¢ JX444594

1039RJP * JX444590

1037RJP * JX444589

1284RJP ¢ JX444591

1308RJP * JX444593

1306RJP ¢ JX444592

246bGRJ JX444644

CMW 22830 4 DQ539536

CMW 22829 4 DQ539535

CMW 22832 4 DQ539538

CMW 22831 4 DQ539537

1132RJ ¢ (=voucher 1138RJ) JQ289021

O. rostrocoronatum CBS 434.77 AY194509

Graphilbum tsugae WIN(M) 1391 *(=UAMH 11701) KJ661745

WIN(M) 1384 ¢ (=UAMH 11700) =

WIN(M) 1387 *(=UAMH 11699) —

Ophiostoma sp.1 WIN(M) 1602 HM363177

WIN(M) 1603 HM363164

Ophiostoma sp. 2 RJ-0771 ! AM943895

RJ-0704 § AM943894

Ophiostoma sp. 3 PR-2006c (=CMW 27315) DQ674367

Graphilbum tsugae sp. nov. (Canada) ... 403

Pesotum fragrans CMW 22853 DQ539561

C01-014a § AY194513

NLC 348 8 AY194518

CBS 279.54 §(=C1224) AF198248

WIN(M) 1388 8 _—

C990 DQ062976

P cupulatum C1194 AF198230

Sporothrix inflata CMW 12529 AY495428

‘Strain numbers followed by identical superscript letters (* to 8) have identical ITS regions and only one

representative was used in the ITS phylogenetic analysis.

*CBS = CBS-KNAW Fungal Biodiversity Center, Utrecht, Netherlands; WIN(M) = culture collection of

J. Reid, Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada; UAMH = University

of Alberta Microfungus Collection and Herbarium, Devonian Botanic Garden, Edmonton, Alberta,

Canada; CMW, Tree Pathology Co-operative Program, Forestry and Agricultural Biotechnology

Institute, University of Pretoria, South Africa; STE-U, Stellenbosch University, South Africa.

period was to have lasted seven days, we incubated all plates further at 20°C to record

morphological development.

DNA extraction, amplification protocols and DNA sequencing

DNA extraction, purification, and agarose electrophoresis protocols followed

Hausner et al. (1992). Genomic DNA served as the amplification template using the

Invitrogen-Life Technologies PCR System (Buffer and Taq polymerase, Invitrogen,

Fredrick, MD). Primers SSUZ and LSU4 (Hausner et al. 2005) were used to amplify the

ITS regions. The PCR primer sequences, amplification conditions, sizes of the expected

PCR products, and preparation of sequencing templates for fragments have been

described previously (Hausner & Wang 2005, Hausner et al. 2005). DNA sequencing

templates were prepared with the aid of the Promega Wizard SV Gel and PCR clean-

up system (Promega, Madison, WI). Purified double-stranded PCR products were

sequenced in both directions using the cycle-sequencing protocols performed according

to the manufacturer's recommendations (Perkin Elmer Applied Biosystems, Foster

City, CA), and automated Fluorescent DNA sequence analysis was performed using

an ABI Prism 310 Genetic Analyzer system (PEAB at the University of Calgary, DNA

sequencing facility, Calgary, AB).

Analyses of DNA sequence data

To add to sequences generated herein, we used the UAMH 11701 ITS sequence in

a BLAST search to recover additional sequences from NCBI databases for comparison

with other relevant taxa. Fifty ITS rDNA sequences were aligned with CLUSTAL-X

(Thomson et al. 1997) and, when appropriate, modified with the alignment editor

program GeneDoc v2.5.010 (Nicholas et al. 1997, http://www.psc.edu/biomed/genedoc).

Phylogenetic trees were generated using the Molecular Evolutionary Genetic Analysis

program package (MEGA 5.1; Tamura et al. 2011). Trees were generated with the

Maximum parsimony (MP), Neighbor joining (NJ), and Maximum likelihood (ML)

methods. MEGA 5.1 was also used for determining the best-fit DNA models and the

best-fit substitution model for ML analysis. In order to evaluate node support within the

404 ... Reid & Hausner

tree topologies, the bootstrap option (1000 replicates) was selected for all tree-building

programs.

Finally, MrBayes (version 3.1; Ronquist & Huelsenbeck 2003) was used for Bayesian

analysis. The NEXUS file format necessary for the alignment (input) file was generated

with the file converting option available within DAMBE (Xia 2001). The DNA

substitution model setting for Bayesian analysis was chosen based on evaluating the

ITS region alignment with the Modeltest 3.7 program (Posada & Crandall 1998). The

Bayesian inference of phylogenies was initiated from a random starting tree, and four

chains were run simultaneously for 3 million generations; trees were sampled every 100

generations. The first 25% of trees generated were discarded (burn-in); the remaining

trees were used to construct a 50 % majority rule consensus tree and compute the

posterior probability values.

Phylogenetic trees were drawn with the TreeView program (Page 1996) and figure

annotations added using Corel Draw (Corel Corporation and Corel Corporation

Limited).

Results

Cultural and morphological characteristics

The temperature-related growth response of isolate VAMH 11701 (Fie. 1)

depicts a relatively broad growth temperature optimum, i.e. 20-25°C, dropping

off sharply towards both 15 and 30°C. The mycelium amount produced differed

between test temperatures but was always appressed, with only limited sparse,

central aerial mycelium. During the first 7 days, the mycelium remained hyaline

(translucent) and without coloration or reproductive structure development.

All eight plate sets from the various test temperatures were then placed in a

single 20°C incubator.

Mean 90

colony 40

diam

0 0

10 15 20 25 30 35

Temperature ( C)

Fic. 1: Temperature-growth curve for Graphilbum tsugae holotype strain UAMH 11701 incubated

at various temperatures for 7 days.

Graphilbum tsugae sp. nov. (Canada) ... 405

By dayl0, mycelium in all original 20°C and 25°C cultures had reached

Petri dish margins. It was still mostly hyaline and appressed, but with a ring

of slightly more floccose white-colored mycelium around the paler center

(Fic. 2A); these cultures were virtually identical in growth rates and appearance.

The 15°C and 30°C cultures had also grown further; their diameters were

still quite small, but the 10°C and 35°C cultures showed only numerous short

hyphae extending from their inoculation discs; apparently the fungus could

survive a short exposure to the relatively high 35°C temperature.

All plate sets were then returned to the 20°C incubator until day 17.

In all original 20°C cultures, the mycelium had completely covered the

dish surfaces. It was still quite peripherally appressed, more floccose centrally,

and while still primarily white, there were definite scattered amber tints

and a central darkening (Fic. 2B). Very occasionally scattered immature

differentiating perithecia were observed, but only a few showed what might

have been developing neck initials.

On day 17 in the original 25°C cultures, the mycelium had also reached dish

margins. Generally it was still white, but now there were scattered slight amber

tints. Less aerial mycelium was present peripherally than centrally, but that

difference was now less pronounced. Overall there was less aerial mycelium

than in the 20°C plates, but the central dark (Fuscous Black) pigmentation

(Fic. 2C) was more pronounced and perithecial development more advanced.

In six dishes the perithecia were quite numerous but fewer in the remaining

two, suggesting that 25°C may be slightly better for perithecial production.

On day 17 in the original 10°C cultures, all isolates were growing, but no

mycelium had reached a dish margin. Colony diameters averaged 70 mm, the

mycelium was white and peripherally appressed to slightly floccose centrally,

but no perithecial initials were present. In the original 15°C cultures, mycelium,

which reached the dish margins randomly, was white, appressed peripherally,

and slightly floccose centrally.

On day 17 in the original 30°C cultures, mycelium had reached the dish

margins uniformly, but again the amounts of central and peripheral mycelium

differed markedly. The peripheral mycelium was white and appressed while

the central mycelium was more floccose and showing numerous presumptive

perithecium initials. And while all original 35°C cultures were growing, no

mycelium had reached a dish margin. Colony diameters averaged 53 mm,

and in every case the mycelium was white, appressed peripherally and more

floccose centrally, and there was no evidence of perithecia.

After inspection, we retained only the original 20°C and 25°C cultures,

which were thereafter both maintained at 20°C.

By day 24, these retained cultures were similar in appearance. New mycelium

was less dense, older mycelium had collapsed centrally, and scattered amber

406 ... Reid & Hausner

Fic. 2. Graphilbum tsugae (holotype, VAMH 11701 [WIN(M) 1391]). Morphology after growth

at various temperatures. A. Original 20 °C plate at day 10. B. Original 20 °C plate at day 17.

C. Original 25 °C plate at day 17. D. Original 25 °C plate at day 24. E. Original 25 °C plate after

c. 7 weeks; F. Original 20 °C plate after c. 7 weeks.

Graphilbum tsugae sp. nov. (Canada) ... 407

colored patches, which were more common, often had turned pale yellowish-

brown. More dark pigment was present centrally in the agar, and mounds of

floccose white hyphae had formed at the colony margins. Although many

perithecial initials were seen, no mature perithecia were found (Fic. 2D, an

original 25°C culture).

Fic. 2E illustrates changes that occurred in an original 25°C culture over

a further period of approximately 6-8 weeks. Note the increased amount of

marginal mounded mycelium, increased area of central dark pigmentation,

slight further overgrowth of superficial aerial mycelium, and finally, what appear

to be abundant immature perithecia. Fic. 2F shows an even older original 20°C

culture with extensive development of — now collapsed — marginal mounded

mycelium but no apparent superficial mycelial overgrowth; there was also a

restricted central dark pigmentation. Numerous perithecia were produced,

but we only rarely saw any evidence of sporulation. Fics. 2E-F both show an

extensive development of cinnamon-colored pigment throughout.

Because we rarely observed fully mature perithecia exuding spore droplets

on agar dishes, we grew the strains on agar plus wood chips where mature

perithecia were readily produced and their nature could be fully characterized

(see TABLE 2 and Taxonomy section).

TABLE 2. Morphological comparisons between G. rectangulisporium and G. tsugae

CHARACTER

PERITHECIAL BASE

Shape

Width (um)

Height (um)

PERITHECIAL NECKS

Length (um)

Base width (um)

Tip width (um)

OSTIOLAR HYPHAE

Length (um)

Number

Shape (um)

ASCOSPORES (uum)

ANAMORPH

COLONY COLOR

Graphilbum rectangulisporium ¢

Globose

70-110

190-500

including ostiolar hyphae

20-35

10-19

13-43

5-15

Cylindrical with apical taper

2.0-3.5 x 1.0-1.7 with sheath;

rectangular in side and face view

None found

White; remaining so on 2% MEA

*Ohtaka et al. 2006; Kirk et al. 2008; ‘Reid et al. 2010.

Graphilbum tsugae

Obpyriform, ampulliform or globose ?;

75-120(-167.5)

75 -113(-140)

(87-)138-200(-280)

including ostiolar hyphae

27.5-50

1225225

(20-)25-40

Various

Crooked to jagged in outline; at times

irregularly cellular; tapering to a point.

3-5 x 1 with sheath;

oblong to rectangular;

sheath flaring at corners

Conidiophores simple to branched; not

identifiable to genus; Conidia hyaline,

1-celled, oblong, 2-5 x 1-1.5 um.

White initially, coloring (pale yellowish

brown) on aging on 2% MEA + 1%

YES

408 ... Reid & Hausner

DNA sequence analysis

The rDNA SSU sequence (GenBank accession: HQ634824) for isolate

UAMH 11701 — then as WIN(M) 1391 — was previously included in a study

examining rDNA SSU intron distributions among ophiostomatoid fungi and

related taxa (Hafez et al. 2012; p. 102). It showed that although UAMH 11701

was part of the Ophiostomataceae clade, it did not group with representatives

of Grosmannia, Ceratocystiopsis or Ophiostoma sensu stricto. Instead, within

the phylogenetic tree it assumed the most basal position among the sampled

Ophiostomataceae taxa.

Our rDNA sequence analysis of the internal transcribed spacer (ITS) region

(ITS1, 5.88, and ITS2; KJ661745) yielded even more surprising results. Using

this sequence set from isolate UAMH 11701 — again as WIN(M) 1391— in

a BLASTN analysis, revealed that its sequences are similar to those from this

Pesotum fragrans AF1982481°

Ophiostoma microcarpum GU134170

19-9 Pesotum fragrans DQ062976

8% | + Ophiostoma sp. PR-2006c DQ674367

109 Ophiostoma microcarpum GU134170

ig Pesotum fragrans DQ539561

; Ophiostoma cf. rectangulosporium Ja289021°

Grap hilbum S) ® ! Ophiostoma cf. rectangulosporium JX444593 ©

Ophiostoma cf. rectangulosporium JX444644

Ophiostoma cf. rectangulosporium EU785451 B

Ophiostoma sp. RJ-0771 AM943895 f

Ophiostoma rectangulosporium GU134171

* Ophiostoma cf. rectangulosporium GU129987

; Ophiostoma rectangulosporium AB242825

Ophiostoma sp. WIN(M)1602 HM363177

Ophiostoma sp. WIN(M)1603 HM363164

az Ophiostoma cf. rectangulosporium GU393357

a Graphilbum Sugee WIN(M)1391°

Ophiostoma grandicarpum AJ2938

@ , Ceratocystiopsis minuta DQ1281 es

Ceratocystiopsis cf. minuta DQ128175

@ Ophiostoma pulvinisporum AY546715

100

100 Ophiostoma ips DQ539549

es @) Ophiostoma piceae EF506934

ie Pesotum cupulatum AF198230

a7 Ophiostoma rostrocoronatum AY 194509

Ophiostoma abietinum DQ396788

55 Sporothrix inflata AY495428

Ophiostoma dentifundum AY495434"

Grosmannia penicillata AM943882

-——_———__ Node support @ = node support >99 for

0.05 values: MB, NJ, MP, ML

Fic. 3: Phylogenetic tree based on ITS sequence data. Levels of confidence for the major nodes

in the phylogenetic tree are based on posterior probability (PP) values (MrBayes; first value), and

bootstrap support (BS) in NEIGHBOR (second value), DNAPARS (third value), and Maximum

likelihood analysis (fourth value). Nodes that received less than 50% support (BS or PP) were

collapsed. Tree topology and branch lengths are based on Bayesian analysis and are proportional to

the number of substitutions per site. GenBank accession numbers are listed next to species names.

Graphilbum tsugae sp. nov. (Canada) ... 409

region in strains identified as Pesotum fragrans, Ophiostoma rectangulisporium,

Ophiostoma cf. rectangulisporium, and O. microcarpum (Fic. 3, TABLE 1).

Comparisons within this ITS data set also strongly suggest that there are

several yet to be described cryptic species within this clade. In particular

the circumscription of O. rectangulisporium and isolates designated as O. cf.

rectangulisporium need to be reevaluated, as the molecular data obtained failed

to group these taxa into an appropriate clade relative to their assigned names.

All these species are currently expected to be treated as members of the now

holomorph genus Graphilbum (see below), whose species form a monophyletic

group apart from species of Ophiostoma sensu stricto.

Taxonomy

Graphilbum tsugae J. Reid & Georg Hausner sp. nov. FIGS. 4-7

MycoBank MB809811

Differs from all previously described species of Graphilbum by its apical ostiolar

projections being not true ostiolar hyphae but elongated tapering neck cells with jagged

outlines, its rDNA sequences, and its variably solitary or sporodochial anamorphic

conidiophores.

Type: Canada, British Columbia, Highway 1, Annis Mountain, near Salmon Arm,

from a felled log of Tsuga heterophylla (Raf.) Sarg., isolated from spore drop at apex

of a perithecium in unidentified bark beetle gallery, 21 September 1987, collected and

isolated by J. Reid. (Holotype, UAMH 11701 [dried culture on agar plus wood chips],

derived from herein designated isotype culture WIN(M) 1391.

EryMo_oey: tsugae, referring to the host genus Tsuga.

Maximum growth occurring on 2% malt-extract agar amended with yeast

extract over a temperature range from 20°C to 25°C; maximum diameter 60

mm after seven days, i.e., relatively slow growing. No measurable growth at

10°C and below or 35°C and above. Colonies remaining only white over this

period.

Anamorph apparently pleomorphic on wood chip agar plates. Some conidia

developing terminally on short, simple, non-branched to slightly branched

conidiophores. Others on variously branched conidiophore clusters that do

not develop in any consistent branching pattern. Some are initially hyaline

and verticillate, but later secondary cymose branches develop (see Seifert et

al. 2011, fig. 100D) arising from beneath a septum in the parent branch. In

others, branches that have arisen initially from a common point continue to

branch along their length in a cymose fashion. Often these variously branched

forms aggregate in clusters on the substrate, and then appear superficially

as sporodochia until mounted. Conidiogenous cells proliferating in an

imprecisely determined manner, as at times either sympodial or percurrent,

or both presumptive forms can be observed in single mounts. Conidia one-

celled, hyaline, oblong and parallel-sided in plan view (see Olchowecki &

410 ... Reid & Hausner

Fic. 4: Graphilbum tsugae (wood chip agar cultures). A. Simple conidiophores bearing conidia.

B. Conidia in plan view (see Olchowecki & Reid 1974, p.1678 for definition). Scale bars: A = 35

um; B = 8 um.

Reid 1974, p. 1678 for definition); generally with slightly rounded ends, but

in some cases one end flattened when fully abscised; very uniform in size,

cylindrical in cross section (arrow); 3-5(-6) (mean + s.d. = 4.0 + 1.05) um long;

frequently produced in slimy masses that occasionally form around the bases of

perithecia developing on the wood chips. In such masses the more complicated

conidiophores appear.

Mature perithecia black; initials produced abundantly on malt extract

supplemented with yeast extract rarely maturing. Abundant mature perithecia

developing in two months on plates containing wood chips embedded in the

agar. Bases obpyriform, ampulliform, or globose, 75-120 (-168) um wide

(mean + s.d. = 102 +16 um), 75-113 (-140) um high (mean + s.d = 94 + 19 um).

Surface smooth to slightly irregular, often adorned with short dark hair-like

hyphae. Neck black, (87-) 138-200 (-280) um long (mean + s.d. = 166 + 49

um), smooth to occasionally hairy; base 27.5-50 um wide (mean + s.d. = 39.1

+ 6.24 um), tapering to 12.5-25 um (mean + s.d. = 18 + 4.0 um) just below the

point of diverging apical projections; these projections are not ostiolar hyphae,

but elongated terminal neck cells, (20-)25-40 um long (mean + s.d. = 33.5 +

7 um), crooked to jagged cellular extensions that taper to a point, apparently

developing by differentiation from normal terminal neck cells; base 2-5 um

Graphilbum tsugae sp. nov. (Canada) ... 411

Fic. 5: Graphilbum tsugae (wood chip agar cultures). Variously branched conidiophores. A. Single

verticillate conidiophore bearing conidia; arising from beneath septum in a parent branch (arrow).

B. Portion of a compound aggregation of conidiophores showing successive cymose branches

(see Seifert et al. 2011, fig.100D) in a single element (arrows). C. Aggregation of verticillate

conidiophores formed in culture on and around the wood chips. Scale bars: A = 12 um; B = 30

um; C = 65 Um.

412 ... Reid & Hausner

Graphilbum tsugae sp. nov. (Canada) ... 413

wide (mean + s.d = 3.5 + 0.7 um). Ascospores hyaline, one-celled, oblong

to rectangular with slightly rounded ends, rarely slightly constricted in the

middle; sheath present but slightly flared and projecting out at the corners in

plan view; 3-5 (mean + s.d. = 3.8 +. 0.7 x 1.0 um).

ADDITIONAL MATERIAL EXAMINED: CANADA, British CoLumBiaA, Highway 1,

Annis Mountain, near Salmon Arm, from a second felled log of Tsuga heterophylla at

the same location from which the holotype material was obtained, 21 September, 1987.

(1) A dried culture grown from isolate WIN(M)1387 (= UAMH 11699] originally

isolated aseptically from a slab of stained wood found adjacent to the beetle galleries

in the log; (2) A dried culture grown from isolate WIN(M)1384 [= UAMH 11700] that

had been isolated from perithecial ooze extruded from a broken perithecium in the

beetle galleries in a second small slab of wood taken from this second log. Both samples

collected and isolated by J. Reid.

Finally, we correct errors in the citation of:

Graphium rectangulisporium (Ohtaka, Masuya & Yamaoka) Z.W.de Beer & M.J.

Wingf. CBS Biodiversity Series 12: 269. 2013 [as “rectangulosporium (R.W.

Davidson)” ].

= Ophiostoma rectangulisporium Ohtaka, Masuya & Yamaoka, Can. J. Bot. 84: 290. 2006

[as “rectangulosporium”; orthographic correction to comply with McNeill et al. 2012:

Art. 60.8].

The miscitation of “R.W. Davidson” as the basionym author appears to have

been a transcriptional error.

Discussion

Kim (2010) and Kim et al. (2011) drew attention to a sterile, white, fast

growing isolate (GU393357) obtained from Orthotomicus erosus (Wollaston)

from Pinus spp. growing in California that groups closely with our new species

(Fic. 3). However, both its reported growth rate and colour range differ

markedly from those of G. tsugae. We believe both our morphological data

and molecular analysis support our isolates as distinct from all other species

formerly assigned to Ophiostoma. They are, however, linked by molecular

criteria to a small group of other former Ophiostoma species whose anamorphs

had been considered members of the then anamorphic genus Graphilbum.

This observation is significant due to recent changes proposed by de Beer et

al. (2013a, figs. 2 & 3, pp. 4 & 6) and de Beer & Wingfield (2013, figs. 4b &

5b, pp. 29 & 31). In both these papers Graphilbum is used as a holomorph-

generic name in trees, and in de Beer & Wingfield (2013), the species group to

be included in Graphilbum is discussed on page 39.

Fic. 6 (left): Graphilbum tsugae (wood chip agar cultures). A-C. Mature perithecia. A. Single

mature perithecium and a detached perithecial neck. B. Cluster of five mature perithecia.

C. Mature perithecia growing on wood chips in agar medium. Note the droplets in which aggregated

conidiophores are often found. D- F. Apical neck projections. Scale bars: A, B = 110 um; C = 650

um; D = 15 um; E= 17 um; F = 10 um.

414 ... Reid & Hausner

Fic. 7 Graphilbum tsugae (wood chip agar cultures). Ascospores. Scale bars: A = 8 um; B = 7 um.

Graphilbum tsugae sp. nov. (Canada) ... 415

De Beer et al. (2013b, p. 268) formally redefined the generic name as:

Graphilbum H.P. Upadhyay & W.B. Kendr., Mycologia 67: 800; 1975 emend.

Z.W. de Beer, Seifert & M.J. Wingf. [type species Gra. sparsum] = Ceratocystis

Ellis & Halst. section Ips H.P. Upadhyay pro parte, Monogr. Ceratocystis and

Ceratocystiopsis, p. 70. 1981).

On page 269 discussing Graphilbum sparsum H.P. Upadhyay & W.B. Kendr.

(1975), de Beer et al. (2013b), also state: ... “Graphilbum sparsum is the type

species of the genus... re-introduced here to accommodate species previously

treated in the P fragrans complex ... ”

Although this resulted in eight species being formally assigned to the genus

Graphilbum — for only six of which phylogenetic data exists — there is now

a ninth: an anamorphic species placed in Graphilbum by phylogenetic data

(Romon et al. 2014). However, it is clear that de Beer and Wingfield (2013,

p. 39) felt strongly that additional isolates studied by them might well deserve

similar placement when further supportive data becomes available. Some

such potential species are listed with former Ophiostoma spp. discussed in

their species review under the name of Graphilbum (de Beer & Wingfield

2013, p. 39; figs. 4 b, p. 29, 5b, p. 31) and also seen in de Beer et al. (2013a,

fig. 3, p. 6). Using phylogenetic data, three other species represented by new

combinations in Hyalorhinocladiella proposed by Harrington et al. (2010) —

H. ips, H. macrospora, H. tingens — were shown to represent anamorphs of

three different Ophiostoma species (de Beer et al. 2013b).

The foregoing changes were made in conformance with the new rule of fungal

nomenclature (McNeill et al. 2012, Article 59, Note 2), “... all legitimate fungal

names are treated equally... regardless of the life history stage of the type... .”

For a history of the origin of this approach see Hawksworth (2011). Following

this practice, we describe our holomorphic isolates as representing a new

species of the genus Graphilbum, originally an anamorphic name as emended

above, even though there are some significant morphological inconsistencies

among the species now included therein. Our new species introduction will

undoubtedly add to these inconsistencies and to the impetus to erect further

new genera.

Our ITS sequence analysis, which showed Graphilbum tsugae only distantly

related to species accommodated in Ceratocystiopsis, Grosmannia, and

Ophiostoma, grouped it in a clade previously referred to as the “P. fragrans &

O. rectangulosporium clade” (Jankowiak 2012). Earlier studies based on ITS

region analyses also showed that P. fragrans and O. rectangulisporium form a

distinct lineage among the Ophiostomatales (Lu et al. 2009, Paciura et al. 2010,

Jankowiak & Kolarik 2010, Jankowiak & Bilanski 2013ab, Romon et al. 2014),

hinting at possible generic status for these taxa. Based on this history, we feel

the ITS data support placing our new species, at least for now, in Graphilbum.

416 ... Reid & Hausner

Because of our ITS data, we critically compared (TABLE 2) our new species

morphologically only to O. rectangulisporium, as described and named in

Ohtaka et al. (2006). This comparison clearly separates G. tsugae from the

only other described Graphilbum species to which it could possibly have been

assigned.

The morphological character that most sets G. tsugae apart from all

other described Graphilbum species are the projections found at the apex of

the perithecial neck; we call these ostiolar projections, for they are certainly

unlike the more typical ostiolar hyphae that develop in many other species of

Ophiostoma sensu stricto.

Ostiolar hyphae are commonly found at the perithecial neck apices of many,

but not all, Ophiostoma species. However, when present, they are generally

easily seen during careful examinations, though in some species they can be

quite short. They may also be quite long, straight or curved or even spirally

curved, parallel-sided or tapered, convergent or divergent, and septate or

aseptate and sometimes may have discrete inflated bases. They often simply

appear to be extensions of the parallel hyphae comprising the perithecial neck

when the neck is so constructed or modified filamentous extensions of the

terminal neck cells that are distinctly cellular in structure.

In G. tsugae, perithecial necks are apparently composed of relatively thick-

walled, primarily oblong to slightly elongate cells. These may also be irregular

in outline, but all abut in an interlocking manner and form serially as necks

elongate (Fic. 6E).

As the elongation phase of the neck body slows, these cells begin to separate

laterally while continuing to elongate apically and tapering to terminate

in a sharp point. To us, they resemble “Zeus’s thunderbolt.” The individual

appendages so formed do not appear to be septate (Fic. 6E). We have never

previously seen such ostiolar ornamentations.

Graphilbum, as emended by de Beer et al. (2013b), will now contain ten

formally described species and, based on rDNA data, several undescribed ones.

Graphilbum species either lack conidial states (O. rectangulisporium, Ohtaka

et al. 2006) or produce anamorphs ranging between hyalorhinocladiella- to

pesotum-like in appearance (de Beer and Wingfield 2013).

Acknowledgments

The authors express their sincere gratitude to Drs. Walter Gams and Tom Grafenhan

for their critical reviews of the manuscript and gratefully acknowledge funding for this

research in part through an operating grant from the Natural Sciences and Engineering

Research Council of Canada. We also thank both Drs. Shaun Pennycook and Lorelei

Norvell for their very helpful suggestions for improvement made during the editorial

phase of this manuscript.

Graphilbum tsugae sp. nov. (Canada) ... 417

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Biodiversity Centre, Utrecht (The Netherlands). Biodiversity Series 12: 21-46.

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Wingfield MJ. (Eds.), CBS-KNAW Fungal Biodiversity Centre, Utrecht (The Netherlands).

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MY COTAXON

http://dx.doi.org/10.5248/130.421

Volume 130, pp. 421-427 April-June 2015

Molecular analyses reveal a new species in Melanoderma

from tropical China

Ha1-SHENG YUAN * & YU-HE KAN”?

"State Key Laboratory of Forest and Soil Ecology, Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang 110164, P. R. China

? Graduate University of the Chinese Academy of Sciences, Beijing 100049, P. R. China

* CORRESPONDENCE TO: hsyuan@iae.ac.cn

ABSTRACT — A new wood-decaying polypore, Melanoderma disciforme, is described from

tropical China based on morphological characteristics and rDNA ITS sequences. The new

species is characterized by perennial effuse-reflexed basidiocarps, a narrow pileus with a thin

black crust on the pileal surface, a dimitic hyphal system with dextrinoid and cyanophilous

skeletal hyphae, and the presence of fusiform cystidioles and cylindrical basidiospores.

Molecular analyses indicate that the new species and M. microcarpum cluster together with

full support.

Key worps —Basidiomycota, Polyporaceae, phylogeny, taxonomy

Introduction

The genus Melanoderma B.K. Cui & Y.C. Dai was recently established

to accommodate the newly described species M. microcarpum B.K. Cui &

Y.C. Dai (Cui et al. 2011). The genus is characterized by perennial pileate

to effuse-reflexed basidiocarps, a black pileal surface with a thin crust, a

dimitic hyphal system with dextrinoid cyanophilous skeletal hyphae, apically

encrusted cystidia, and the presence of cystidioles and cylindrical thin-walled

basidiospores. Microscopically, Melanoderma resembles Perenniporia Murrill

in having dextrinoid and cyanophilous skeletal hyphae. However, Melanoderma

is distinguished from Perenniporia by its distinctly thin-walled, non-dextrinoid,

and acyanophilous basidiospores (Cui et al. 2011).

China is very rich in wood-inhabiting fungi species, and recently extensive

studies have been conducted on species diversity, taxonomy, ecological

patterns, and phylogeny of wood-inhabiting fungi (Dai 2010, 2011, 2012, Dai

et al. 2007, Ma et al. 2011, Zhang et al. 2009, Zhou & Dai 2012). Two polypore

specimens collected in 2005 from a tropical forest of Yunnan Province were

422 ... Yuan & Kan

subsequently identified and deposited as Melanoderma microcarpum. But

recent comprehensive morphological and molecular analyses suggest that these

specimens represent an undescribed Melanoderma species. Here we clarify its

taxonomic status and describe it as a new species.

Materials & methods

Morphological studies

The specimens studied are deposited in the herbarium of the Institute of Applied

Ecology, Chinese Academy of Sciences, Shenyang, China (IFP). The microscopic

procedure follows He & Dai (2012). Sections were studied at magnifications up to x1000

using a Nikon Eclipse 80i microscope and phase contrast illumination. Measurements

were made from sections stained with CB. In presenting basidiospore size variation, the

5% of the measurements excluded from each end of the range are shown in parentheses.

The meanings of abbreviations are as follows: CB = cotton blue, CB- = acyanophilous,

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

hydroxide, L = mean basidiospore length (arithmetic average of all basidiospores),

W = mean basidiospore width (arithmetic average of all basidiospores), Q = range of

L/W ratios from individual specimens, and n = number of basidiospores measured from

given number of specimens. Special color terms follow Petersen (1996).

Phylogenetic analysis

Total DNA was extracted from specimens using Phire® Plant Direct PCR Kit

(Finnzymes, Finland). Primers ITS5 and ITS4 (White et al. 1990) were used to amplify

ITS sequences following PCR procedure set forth by Stéger et al. (2006). The DNA was

sequenced at Beijing Genomics Institute. The most similar sequences were downloaded

from GenBank NCBI (http://www.ncbi.nlm.gov) using the BLAST tool. Sequences

were aligned using ClustalX (Thomson et al. 1997) and manually edited to allow

maximum alignment and minimize gaps. Maximum parsimony and Bayesian analysis

were applied to the ITS dataset. All characters were weighted and gaps were treated as

missing data. Maximum parsimony analysis were carried in PAUP* (version 4.0b10;

Swofford 2002). Trees were inferred using the heuristic search option with TBR branch

swapping and 1,000 random sequence additions. Max-trees were set to 5000 and no-

increase, branches of zero length were collapsed, and all parsimonious trees were saved.

Nodal support was tested with bootstrap of 1000 replicates using the heuristic search

option (TBR and MULTREES options on) (Felsenstein 1985). Descriptive tree statistics

tree length (TL), consistency index (CI), retention index (RI), rescaled consistency

index (RC), and homoplasy index (HI) were calculated for all trees generated under

different optimality criteria. Bayesian analysis was carried in MrBayes3.1.2 (Ronquist &

Huelsenbeck 2003) and models of evolution were identified using MrMODELTEST2.3

(Posada & Crandall 1998, Nylander 2004). Four simultaneous Markov chains were

run starting from random trees and keeping one tree every 100th generation until the

average standard deviation of split frequencies was below 0.01. The value of burn-in

was set to discard 25% of trees when calculating the posterior probabilities. Bayesian

posterior probabilities were obtained from the 50% majority rule consensus of the trees

kept as support for nodes.

Melanoderma disciforme sp. nov. (China) ... 423

100/1.00 | Melanoderma disciforme Yuan1643

100/1.00 Melanoderma disciforme Yuan1675

100/1.00 HQ678173 Melanoderma microcarpum

HQ678174 Melanoderma microcarpum

100/1.00 } AF511439 Polyporus tubaeformis

HQ604799 Polyporus badius

100/1.00 | JN165002 Datronia mollis

AB587623 Datronia mollis

-/0.90 | KJ140736 Datronia stereoides

79/1.00 JQ673031 Datronia scutellata

100/1.00 ‘ A

KC415178 Datronia stereoides

AB587637 Pseudofavolus cucullatus

AF516570 Polyporus mikawai

JX968555 Pyrofomes demidoffii

JQ861754 Perenniporia amazonica

JX840347 Ganoderma fornicatum

HQ848473 Perenniporia tephropora

HQ654106 Perenniporia tephropora

HQ896245 Dichomitus albidofuscus

100/1.00! EJ340897 Dichomitus albidofuscus

100/1.00

-/0.54

-/0.57 82/0.63

100/1.00 HQ876603 Perenniporia martia

FJ411093 Perenniporia martia

96/0.84 JN411113 Grammothele denticulata

JN411115 Grammothele lineata

100/1.00! Jn1411116 Grammothele lineata

JN411114 Grammothele fuligo

JQ247978 Polyporus tricholoma

100/1.00 | JQ780384 Megasporoporiella subcavernulosa

100/1.00 JQ780383 Megasporoporiella subcavernulosa

100/1.00 | JQ314355 Megasporoporiella rhododendri

JQ780388 Megasporoporiella rhododendri

JQ314377 Megasporporiella lacerata

JQ780412 Dichomitus eucalypti

FN907906 Cinereomyces lindbladii

54/-

100/-

60/0.84

—10.0

Fic. 1. Strict consensus tree showing the phylogeny of Melanoderma disciforme and related species

generated by Maximum Likelihood and Bayesian analysis based on ITS sequences. Parsimony

bootstrap values >50% and Bayesian posterior probabilities >0.50 are shown on the branches.

Phylogenetic results

Two ITS sequences were newly generated in this study (GenBank accession

numbers KM521268 and KM521269). The ITS dataset contains 34 sequences

representing 24 species, among which Cinereomyces lindbladii (Berk.) Jiilich

was selected as outgroup. The alignment comprised 601 characters, of which

297 were constant, 50 were variable but parsimony-uninformative, and 254

were parsimony-informative. Maximum parsimony analysis yielded one

parsimonious tree (CI = 0.505, RI = 0.669, RC = 0.338, HI = 0.495). Bayesian

analysis ran 5 million generations and resulted in average standard deviation

of split frequencies = 0.007737. The 50% majority consensus tree generated by

the Bayesian analysis showed a similar topology with the strict consensus MP

tree. The strict consensus tree is shown in Fic. 1. Both bootstrap support value

(BP) and Bayesian posterior probabilities (BPP) are shown at the nodes. In the

phylogenetic tree, two sequences of M. disciforme were grouped together with

full support (100% BP, 1.00 BPP), and formed a monophyletic lineage with

M. microcarpum fully supported (100% BP and 1.00 BPP).

424 ... Yuan & Kan

Taxonomy

Melanoderma disciforme H.S. Yuan, sp. nov. Fic. 2

MycoBank MB 810703

Differs from Melanoderma microcarpum by its bigger pores, absence of encrusted

cystidia, distinctly smaller cystidioles, and shorter basidiospores.

Type: China. Yunnan Prov., Xishuangbanna, Jinghong County, Nabanhe Nat. Res.,

on fallen angiosperm branch, 15.VHI.2005, Yuan 1675 (holotype, IFP; GenBank ITS,

KM521269).

EryMo_oey: the epithet “disciforme” refers to the disk-like basidiocarps.

BASIDIOCARPS: Perennial, effuse-reflexed, growing as round pulvinate patches

at first, then fusing laterally, margin narrowly reflexed as a pileus, back attached

on substrates, coriaceous when fresh, woody hard upon drying, without odor

or taste when fresh; pileus up to 3 mm broad, 5 cm long, and 3 mm thick at

base. Pileal surface cream to buff when young, becoming black with a thin crust

when aged, concentrically zonate and sulcate, glabrous; margin obtuse, cream

to buff, <0.5 mm wide. Pore surface white to cream when fresh, buff when dry;

pores round to angular, 6-7 per mm, dissepiments thick, entire; sterile margin

distinct, <0.3 mm wide. Context cream to buff, hard corky, <1 mm thick. Tubes

cream to buff, corky, distinctly stratified, about 1 mm long for each layer.

HYPHAL STRUCTURE: Hyphal system dimitic; generative hyphae clamped;

skeletal hyphae dextrinoid and strongly cyanophilous; tissue unchanged in

KOH.

CONTEXT: Generative hyphae scarce, hyaline, thin-walled, rarely branched,

1.5-2.5 um in diam; skeletal hyphae dominant, hyaline, thick-walled to almost

solid, frequently branched, interwoven, 1.5-3 um in diam.

TUBES: Generative hyphae scarce, hyaline, thin-walled, rarely branched,

2.2-3 um in diam; skeletal hyphae dominant, hyaline, thick-walled to almost

solid, occasionally branched, interwoven, 1-3.5 um in diam. Cystidia absent;

cystidioles fusiform, hyaline, thin-walled, 11-14 x 3-4 um. Basidia clavate to

barrel, with four sterigmata and a basal clamp connection, 10-13 x 5-6 um;

basidioles similar in shape to basidia, but slightly smaller. Polygonal crystals

frequently present in trama.

Basiprospores: Cylindrical, hyaline, thin-walled, smooth, IKI-, CB-,

(4.8-)4.9-5.3(-5.5) x (1.9-)2-2.3(-2.5) um, L = 5.05 um, W = 2.17 pm,

Q’= 2.32+2.34 (n = 34/2).

TYPE OF ROT: Causes a white rot in angiosperms.

ADDITIONAL SPECIMENS EXAMINED: Melanoderma disciforme: CHINA. YUNNAN

Prov., Xishuangbanna, Jinghong County, Nabanhe Nat. Res., fallen angiosperm branch,

15.VIII.2005, Yuan 1643 (IFP; GenBank ITS: KM521268).

Melanoderma microcarpum: CHINA. HUNAN PRov., Yizhang County, Mangshan

Forest Park, on fallen angiosperm trunk, 25.VI.2007, Dai 8116 (BJFC, holotype).

Melanoderma disciforme sp. nov. (China) ... 425

es §0000000

a: a

5 um

10 pm

Fic. 2. Melanoderma disciforme microscopic structures (Yuan 1675, holotype). a. Basidiospores;

b. Basidia and basidioles; c. Cystidioles; d. Hyphae from trama; e. Hyphae from subiculum.

Discussion

Melanoderma disciforme represents the second species in the genus. It is

characterized by perennial effuse-reflexed basidiocarps, narrow pileus with

a thin black crust on the pileal surface, fusiform cystidioles, and cylindrical

426 ... Yuan & Kan

basidiospores. The molecular phylogeny and morphological evidences support

it as a new species.

Melanoderma disciforme shares most of the morphological features with

M. microcarpum, the type species of the genus, which differs by smaller pores

(7-9 per mm), apically encrusted cystidia, distinctly larger cystidioles, and

longer basidiospores.

Species in Perenniporia, Megasporoporiella B.K. Cui et al., Datronia Donk,

and Dichomitus D.A. Reid are morphologically similar to M. disciforme by

having a dimitic hyphal system with cyanophilous skeletal hyphae. However,

Perenniporia species have thick-walled basidiospores, Megasporoporiella

species lack a crust on the surface of the pileus, and Datronia and Dichomitus

species have non-dextrinoid skeletal hyphae (Nufez & Ryvarden 2001, Li & Cui

2013). Additionally, phylogenetic analyses based on ITS sequences indicate that

species sampled from these genera are not genetically close to M. disciforme.

Acknowledgements

The authors would like to express their deep thanks to Drs. Juliano Marcon Baltazar

(Universidade Estadual de Maringa, Brazil) and Shuanghui He (Beijing Forestry

University, China) for serving as pre-submission reviewers. This research was financed

by the National Natural Science Foundation of China (Project Nos. 31170022 &

31470148).

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MY COTAXON

http://dx.doi.org/10.5248/130.429

Volume 130, pp. 429-436 April-June 2015

Graphis hongkongensis sp. nov.

and other Graphis spp. new to Hong Kong

WEI GUO *? & JAE-SEOUN HurR‘*

'Korean Lichen Research Institute, Sunchon National University, Suncheon 540-950, Korea

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

* CORRESPONDENCE TO: jshur1@sunchon.ac.kr

ABSTRACT — Eleven Graphis species are newly recorded from Hong Kong. Graphis

hongkongensis is new to science, G. centrifuga and G. hyphosa are new to China, and

G. assimilis, G. descissa, G. duplicata, G. immersella, G. immersicans, G. japonica, G. parallela,

and G. plagiocarpa are new to Hong Kong. Graphis hongkongensis possesses lineola-morph

lirellae, a completely carbonized exciple, small to medium-sized ascospores, and the presence

of stictic acid. A brief description of each species is given.

Key worps — Graphidaceae, lichenized fungi, new species

Introduction

Graphidaceae, with over 2300 species belonging to more than 50 genera,

is considered the largest crustose lichen community (Lucking et al. 2009;

Rivas Plata et al. 2010, 2012; Rivas Plata & Liicking 2013). Graphis, its type

genus, is characterized by lirelliform ascomata; a carbonized proper exciple;

a hymenium inspersed with granules or not; hyaline, transversely septate to

muriform, amyloid ascospores reacting blue or blue-violet in iodine; and the

presence or absence of lichen compounds. There are more than 70 Graphis

species in China (Jia & Wei 2008, 2009, 2011; Jia 2011, Jia et al. 2012), of which

18 taxa have been reported previously in Hong Kong (Thrower 1988, Wei 1991,

Aptroot & Seaward 1999, Aptroot & Sipman 2001, Seaward & Aptroot 2005).

A revision of Graphis from Hong Kong conducted under the frame of the

Flora Lichenum Sinicorum resulted in the discovery of 11 new records for

Hong Kong, including a new species, Graphis hongkongensis, and two new

records for China, G. centrifuga and G. hyphosa.

430 ... Guo & Hur

Materials & methods

Hong Kong, special Administrative Region of the People’s Republic of China, is located

at the southern coast of China with subtropical climate. Material was collected mainly

from Hong Kong in 2011 and is preserved in the Herbarium Mycologicum Academiae

Sinicae-Lichenes (HMAS-L). A Motic SMZ-168 dissecting microscope (Scientific

Instrument Company, Inc., Campbell, USA) and Zeiss Axioskop 2 plus compound

microscopes (Zeiss Scope, Oberkochen, Germany) in part connected to Nikon D50

digital microscope cameras (Nikon, Tokyo, Japan) were used for the morphological

and anatomical examinations. Lugol’s solution, spot tests, and TLC in solvent system C

(toluene : acetic acid = 200 : 30) were used for chemical study (Culberson & Kristensson

1970, Culberson 1972, White & James 1985).

Taxonomy

Graphis assimilis Nyl., Bull. Soc. Linn. Normandie, sér. 2, 2: 109. 1868.

Thallus, off-white. Ascomata, sinuous, immersed to erumpent, simple to

branched, sometimes clustered (lineola-morph); labia, entire proper exciple,

completely carbonized; hymenium, not inspersed; ascospores 8/ascus,

transversely 8-10-locular, hyaline, 35-50 x 7.5-8.75 um, I+ blue.

CuHEMiIstTRy: Norstictic acid detected in TLC.

DISTRIBUTION & ECOLOGY: Eastern palaeotropical (Licking et al. 2009);

new to Hong Kong. This corticolous species was previously known from China

(Jia & Wei 2011) and India (Adawadkar & Makhija 2006, 2007).

SPECIMEN EXAMINED: HONG KONG. LAUTAU ISLAND, the road from Lautau Peak to

Nam Shan, 22°14’N 113°56’E, alt. 384 m, on bark of tree, 15 Dec. 2011,Wei Guo &

Jinxiu Tian HK367.

REMARKS: ‘This species is anatomically and chemically similar to Graphis

intricata Fée, which differs in its deserpens-or centrifuga-morph lirellae and

smaller ascospores (15-30 um long).

Graphis centrifuga Rasanen, Suom. Elain-ja Kasvit. Seuran Van. Tiedon. Péytakirjat

3: 186. 1949.

Thallus, yellowish grey. Lirellae, immersed to sessile, in satellite clusters

(centrifuga-morph); proper exciple, completely carbonized; hymenium,

100-120 um high, inspersed; ascospores, 8/ascus, transversely 6—8-locular,

hyaline, 20-25 x 5-10 um, I-.

CuHEMIsTRY: Norstictic acid detected in TLC.

DISTRIBUTION & ECOLOGY: Eastern palaeotropical (Licking et al. 2009);

new to China. Graphis centrifuga is a corticolous species known also from

Australia (Staiger 2002).

SPECIMEN EXAMINED: HONG KONG, NEw TERRITORIES, Ma On Shan, Mui Tsz Lam,

22°23’N 114°14’E, alt. 187 m, 14 Dec. 2011, Wei Guo HK358.

Graphis hongkongensis sp. nov. (China) ... 431

REMARKS: Graphis centrifuga most closely resembles G. cervinonigra Zahlbr.,

which differs in its short lirellae (coarctata-morph).

Graphis descissa Mill. Arg., Bull. Herb. Boissier 3: 318. 1895.

Thallus, off-white. Labia entire; lirellae immersed (subserpentina-morph);

proper exciple completely carbonized; hymenium 100-120 um high, not

inspersed; ascospores 8/ascus, transversely 8-10-locular, (20-)28-37 x

6.25-8 um, I+ blue.

CHEMISTRY: Stictic acid.

DISTRIBUTION & ECOLOGY: Neotropical and eastern palaeotropical (Liicking

et al. 2009); new to Hong Kong. The species is an inconspicuous, corticolous

species that also occurs in Australia (Archer 2006).

SPECIMEN EXAMINED: HONG KONG, NEw TERRITORIES, Ma On Shan, Mui Tsz Lam,

22°23’N 114°14’E, alt. 187 m, 14 Dec. 2011, Wei Guo HK356-1.

REMARKS: Graphis descissa resembles G. flavovirens Makhija & Adaw. in its

thallus containing stictic acid, but the lirellae in G. flavovirens are very long and

radially branched (centrifuga-morph).

Graphis duplicata Ach., Syn. Meth. Lich.: 81. 1814.

Thallus, whitish grey to grey. Labia striate (striatula-morph); proper exciple

laterally carbonized; ascospores 8/ascus, transversely 8-9-locular, hyaline,

32.5-42.5 x 5-7.5 um, I+ blue.

Cuemistry: No lichen compounds detected in TLC.

DISTRIBUTION & ECOLOGY: Pantropical (Licking et al. 2009); new to Hong

Kong. Graphis duplicata is a widely distributed species, found on tree bark

in China (Jia & Wei 2011), India (Adawadkar & Makhija 2007), and South

America, Philippines, Indonesia, and Australia (Staiger 2002, Archer 2006).

SPECIMEN EXAMINED: HONG KONG, NEw TERRITORIES, Ma On Shan, Mui Tsz Lam,

22°23’N 114°14’E, alt. 70 m, 14 Dec. 2011, Wei Guo HK349-3.

REMARKS: Graphis duplicata can be distinguished from G. tenella Ach. in

having striatula-morph lirellae and from G. striatula (Ach.) Spreng. in having

smaller ascospores.

Graphis hongkongensis Wei Guo & J.S. Hur, sp. nov. PLATE 1

FUNGAL NAME FN570089

Differs from Graphis gloriosensis by its lineola-morph lirellae and its smaller ascospores

with less trans-septation.

TypE- China, Hong Kong, Lautau Island, the road from Lautau Peak to Nam Shan, on

bark, 22°14’N 113°56’E, alt. 384 m, 15 Dec. 2011, W. Guo & J.X. Tian HK365 (Holotype,

HMAS-L 128226).

EryMoLocy- referring to the type locality, Hong Kong.

432 ...Guo & Hur

PLaTE 1. Graphis hongkongensis (holotype, HMAS-L 128226). A, habit; B, cross-section of an

apothecium; C, ascus with ascospores; D, ascospores.

THALLUS pale grayish white to pale white, smooth. Ascomata lirelliform, sub-

immersed to erumpent, simple to sparsely branched, 1-4 mm long, 0.2-0.3

mm wide. Disc concealed (narrow to slightly open), blackish brown to black,

without pruina. Thalline margin lateral, well developed (lineola-morph lirellae).

Proper exciple completely carbonized. Hymenium colourless, inspersed,

90-135 um high, I-. Paraphyses unbranched, dense, thin, 1.25-2.5 um wide,

filiform, warty in tips.

Asci 6-8-spored, 82.5-100 x 17.5-25 um. Ascospores fusiform, transversely

8-12-locular, 25-50 x 7.5-10 um, I+ violet-blue.

CHEMISTRY: Stictic acid detected in TLC.

REMARKS: Graphis hongkongensis closely resembles G. gloriosensis A.W. Archer

& Elix in having entire labia, a lateral thalline margin, a completely carbonized

excipulum, an inspersed hymenium, transversely septate ascospores, and

a thallus containing stictic acid. However, G. gloriosensis is distinguished by

marginata-morph lirellae, comparatively larger ascospores (50-90 x 9-16 um)

with more trans-septation, and being so far known only from Australia

(Licking et al. 2009).

Graphis kelungana Zahlbr., which resembles G. hongkongensis in having

entire labia, an inspersed hymenium, transversely septate and similarly

sized ascospores, a thallus containing stictic acid, and a similar geographical

Graphis hongkongensis sp. nov. (China) ... 433

distribution, is clearly separated by its lateral carbonization and subserpentina-

morph lirellae. Graphis lineola Ach., which has similar lirellae morphology and

shares most of the anatomical characteristics with G. hongkongensis, differs by

its laterally carbonized excipulum and a thallus lacking lichen compounds.

Graphis hyphosa Staiger, Bibl. Lichenol. 85: 235. 2002.

Thallus, light yellow to yellowish grey. Ascomata immersed in

pseudostromata, stellately branched (hyphosa-morph); labia entire; proper

exciple completely carbonized; hymenium clear; ascospores 4-8/ascus,

transversely 6-9-locular, hyaline, 20-37.5 x 7.5-10 um, I+ blue.

CueEmistry: No lichen compounds detected in TLC.

DISTRIBUTION & ECOLOGY: Neotropical (Licking et al. 2009). New to China.

Graphis hyphosa occurs on tree bark in Costa Rica and Brazil (Staiger 2002).

SPECIMEN EXAMINED: HONG KONG, Lautau ISLAND, the road from Lautau Peak to

Nam Shan, 22°14’N 113°56’E, alt. 384 m, 15 Dec. 2011, Wei Guo & Jinxiu Tian HK370.

REMARKS: Graphis intricata, which also has entire labia, a completely carbonized

excipulum, a clear hymenium, and transversely septate ascospores, can easily be

distinguished from G. hyphosa by its deserpens- or centrifuga-morph, smaller

ascospores, and thallus containing norstictic acid.

Graphis immersella Mill. Arg., Bull. Herb. Boissier 3: 319. 1895.

Thallus off-white. Ascomata lirellae, entire labia, lineola-morph; proper

exciple, laterally carbonized; hymenium, not inspersed; ascospores 8/ascus,

transversely 6-11-locular, hyaline, 20-47.5 x 7.5-12.5 um, I+ blue.

CHEMISTRY: Stictic acid detected in TLC.

DISTRIBUTION & ECOLOGY: Palaeotropical (Liicking et al. 2009); new to

Hong Kong. The species is a corticolous species known from China (Jia &

Wei 2011, Zahlbruckner 1930); the Solomon Islands, Christmas Island, and

Vanuatu (Archer 2006); Australia (Archer 1999); and India (Adawadkar &

Makhija 2007).

SPECIMEN EXAMINED: HONG KONG, New TERRITORIES, Tai Mo Shan, 22°24’N

114°54’E, alt. 906 m, 11 Dec. 2011, Meng Liu HK218.

REMARKS: Graphis immersella resembles G. leptogramma Nyl. in ascomatal

anatomy and a thallus producing stictic acid but differs in its slightly smaller

ascospores.

Graphis immersicans A.W. Archer, Aust. Syst. Bot. 14: 262. 2001.

Thallus, whitish grey. Ascomata lirellae, erumpent (lineola-or deserpens-

morph); proper exciple, completely carbonized; hymenium 70-90 um high,

not inspersed; ascospores, 4—6/ascus, transversely 6-12-locular, hyaline, 25-45

x 7.5-10 um, I+ blue.

434 ... Guo & Hur

Cuemistry: No lichen compounds detected in TLC.

DISTRIBUTION & ECOLOGY: Pantropical (Licking et al. 2009); new to Hong

Kong. This corticolous species was previously reported from China (Jia & Wei

2011), Australia (Archer 2006), and Philippines (Licking et al. 2008)

SPECIMEN EXAMINED: HONG KONG, NEw TERRITORIES, Ma On Shan, Mui Tsz Lam,

22°23’N 114°14’E, alt. 187 m, 14 Dec. 2011, Wei Guo HK359.

REMARKS: Graphis immersicans is close to G. immersella in lirellate morphology,

but G. immersella can be distinguished by its laterally carbonized excipulum

and thallus producing stictic acid.

Graphis japonica (Mill. Arg.) A.W. Archer & Liicking, Lichenologist 41: 437. 2009.

Thallus, green to greyish green. Ascomata lirellae, subserpentina-morph;

proper exciple apically to laterally carbonized; hymenium, 140-160 um high,

not inspersed; ascospores, 4/ascus, muriform, 10-13/2-4-locular, hyaline,

30-67.5 x 12.5-20 um, I+ blue.

CHEMISTRY: Stictic acid detected.

DISTRIBUTION & ECOLOGY: Eastern palaeotropical (Licking et al. 2009);

new to Hong Kong. Graphis japonica predominantly grows on trees and has

been previously reported from China (Lamb 1963, Wang-Yang & Lai 1973, Jia

& Wei 2011) and Japan (Nakanishi 1966, Nakanishi et al. 2003).

SPECIMENS EXAMINED: HONG KONG, NEw TERRITORIES, Tai Po Kau Country Park,

Blue Road, 22°25’N 114°10’E, alt. 136 m, 10 Dec. 2011, Jinxiu Tian HK047, HK052; the

road from Nam Chung to Bride Pool, 22°38’N 114°12’E, alt. 41 m, 9 Dec. 2011, Xinli

Wei & Meng Liu HK009.

REMARKS: This species is most similar to Graphis streblocarpa (Bél.) Nyl. in all

morpho-anatomical and chemical characteristics, but G. streblocarpa has larger

ascospores and 1-2-spored asci.

Graphis parallela Mill. Arg., Nuovo Giorn. Bot. Ital. 29: 200. 1892.

Thallus, whitish grey to ivory-white. Ascomata lirellae opegrapha-morph;

labia entire; proper exciple completely carbonized; hymenium, not inspersed;

ascospores 2-4/ascus, transversely 16-23-locular, hyaline, (35-)65-112.5 x

(7.5-)10-15 um, I+ blue.

Cuemistry: No lichen compounds detected.

DISTRIBUTION & ECOLOGY: Eastern palaeotropical (Licking et al. 2009);

new to Hong Kong. This corticolous species was previously reported from

China (Jia & Wei 2011) and Japan (Nakanishi 1966).

SPECIMEN EXAMINED: HONG KONG, New TERRITORIES, Ma On Shan, Mui Tsz Lam,

22°23’N 114°14’E, alt. 184 m, 14 Dec. 2011, Meng Liu HK269.

REMARKS: Graphis parallela is nearest to G. bifera Zahlbr., which differs by its

considerably shorter, unbranched sessile, opegrapha-morph lirellae.

Graphis hongkongensis sp. nov. (China) ... 435

Graphis plagiocarpa Fée, Essai Crypt. Ecorc.: 38. 1825.

Thallus off-white to greyish white. Ascomata lirellae, dussii-morph; labia

entire; proper exciple completely carbonized; hymenium, not inspersed;

ascospores, 2/ascus, muriform, hyaline, (20-)25-28/1-5-locular, 90-150 x

(25-) 40-50 um, I+ blue.

Cuemistry: No lichen compounds detected.

DISTRIBUTION & ECOLOGY: Pantropical (Lticking et al. 2009); new to Hong

Kong. G. plagiocarpa is a corticolous species previously recorded from China

Jia & Wei 2011), Japan (Nakanishi 1966; Yoshimura 1974), Europe (Staiger

2002) and Australia (Archer 2005).

SPECIMENS EXAMINED: HONG KONG, NEw TERRITORIES, Tai Po Kau Country Park,

Blue Road, 22°25’N 114°10’E, alt. 157 m, 10 Dec. 2011, Meng Liu HK041-2; Kadoori

Garden, Kwun Yam Shan, 22°25’N 114°07’E, alt. 550 m, 12 Dec. 2012, Meng Liu HK242,

HK237.

REMARKS: Graphis plagiocarpa is very similar to G. lumbschii (A.W. Archer)

A.W. Archer in lirellate morphology but differs in lacking lichen compounds.

Graphis cleistoblephara Nyl. and G. sorsogona Vain. also have short unbranched

lirellae (dussii-morph), but G. cleistoblephara produces norstictic acid, and G.

sorsogona has a laterally carbonized exciple.

Acknowledgments

This work was supported by a grant from the Korean National Research Resource

Center Program (NRF, 2011-0031494). The authors are indebted to Prof. Dr. J. C. Wei,

in providing precious suggestions and help in this study. The authors are grateful to Dr.

Ze-Feng Jia and Dr. Santosh Joshi for their valuable comments on the manuscript. We

express our thanks to senior technician Ms H. Deng for giving assistance in HMAS-L,

to Dr. X.L. Wei, Ms J. X. Tian, and Mr. M. Liu in helping specimen collection, and to Dr.

H.J. Liu for modification of the graphics.

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MYCOTAXON

http://dx.doi.org/10.5248/130.437

Volume 130, pp. 437-443 April-June 2015

Two new conidial fungi from Chapada Diamantina, Brazil

CAROLINA RIBEIRO SILVA', Luis FERNANDO PASCHOLATI GUSMAO? ,

& RAFAEL F. CASTANEDA-RuIz?

' Depto. de Micologia, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil

*Depto. de Ciéncias Biologicas, Lab. de Micologia, Universidade Estadualde Feira de Santana,

Avenida Transnordestina s/n, bairro Novo horizonte, 44036-900, Feira de Santana, Brazil

*Inst. de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

* CORRESPONDENCE TO: /gusmao@uefs. br

ABSTRACT —Phaeoschizotrichum ramosum gen. & sp. nov. and Pyriculariopsis bicolorata sp.

nov., two new asexual fungi collected on leaves of Calophyllum brasiliense in the Brazilian

semiarid Caatinga region, are described and illustrated. Phaeoschizotrichum ramosum is

characterized by distinctly branched conidiophores, discrete, terminal, sympodially extended

conidiogenous cells, and subulate to narrowly obclavate, 1-4-septate, pale brown conidia.

Pyriculariopsis bicolorata is distinguished by conidia that are obclavate to subulate, 1-septate,

bicolored, and smooth.

Key worps — microfungi, taxonomy, leaf litter

Introduction

During an investigation of conidial fungi associated with decaying leaves

of Calophyllum brasiliense Cambess. (Clusiaceae) in a riparian forest of

Chapada Diamantina (Piata, Bahia) in the semiarid Caatinga region of Brazil,

two interesting fungi were collected. One was remarkably different from all

previously described hyphomycete genera (Seifert et al. 2011) while the other

differed from all Pyriculariopsis species (Iturriaga et al. 2008, Soares et al. 2011).

The two fungi are therefore described here as new.

Materials & methods

During studies in periods September 2013 to July 2014 leaves of C., brasiliense were

collected in Piata, Bahia. The material was taken to the laboratory using the methodology

by Castaneda-Ruiz (2005). Sample leaves of C. brasiliense were placed in plastic bags. In

438 ... Silva, Gusmao, & Castafieda-Ruiz

the laboratory the samples were washed, placed in Petri dish moist chambers, and stored

in a polystyrene box with sterile water plus glycerol for 30 days. Slide 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

microscope (model BX51) equipped with bright field and Nomarski interference optics.

The type specimens are deposited in the Herbarium of Universidade Estadual de Feira

de Santana, Brazil (HUEFS).

Taxonomy

Phaeoschizotrichum C.R. Silva, Gusmao & R.F. Castafieda, gen. nov.

MycoBank MB810712

Differs from Schizotrichum by its mostly branched conidiophores, its percurrent and

sympodially extended small denticulate and slightly melanized conidiogenous cells, and

its pale brown to brown conidia.

TYPE SPECIES: Phaeoschizotrichum ramosum C.R. Silva et al.

Erymo.ocy: Greek, Phaeo-, meaning dark, referring to the pale brown conidial

pigmentation; Latin, -schizotrichum referring to the hyphomycete genus Schizotrichum.

Cotontss hairy, effuse, brown. Mycelium superficial. ConIpIoPHORES distinct,

single, mostly branched above, septate, brown to dark brown. CONIDIOGENOUS

CELLS polyblastic, denticulate, integrated or discrete, enteroblastic percurrent

elongated and sympodial extensions, terminal. Conidial secession schizolytic.

Conliplia solitary, subulate, narrow obclavate to subcylindrical, euseptate, pale

brown to brown, smooth or verruculose.

Note: Phaeoschizotrichum superficially resembles Schizotrichum McAlpine

(Ellis 1971, Seifert et al. 2011), but Schizotrichum has immersed mycelium and

forms erumpent dark brown textura angularis stromata; setae are sometimes

present and unbranched, dark brown, and thick-walled while its conidiophores

are caespitose, unbranched, and bear sympodial elongated conidiogenous cells

and subulate or filiform, smooth, hyaline conidia. Phaeoschizotrichum also

superficially resembles other cercosporoid fungi with pale or dark pigmented

conidia that are known to cause foliar diseases, such as Passalora Fr. and

Pseudocercospora Speg.

Phaeoschizotrichum ramosum C.R. Silva, Gusmao & R.F. Castafieda, sp. nov.

MycoBank MB810713 FIGS 1, 2

Differs from Schizotrichum lobeliae by its superficial mycelium composing an

anastomosed network of hyphae with a textura epidermoidea appearance, its mostly

branched multi-septate ventricose conidiophores, and its pale brown conidia.

Type: Brazil, Bahia, Piata, Serra da Tromba, 13°07’S 41°50’ W, on dead leaf of Calophyllum

brasiliense, 19 XI. 2013, coll. C.R. Silva (Holotype: HUEFS 210423).

ErymMo_oey: Latin, ramosum, referring to the branched conidiophores.

Phaeoschizotrichum gen. & sp. nov. & Pyriculariopsis sp. nov. ... 439

—

10 um

Fic. 1. Phaeoschizotrichum ramosum (ex HUEFS 210423). A-D. Conidia. E-H. Conidiogenous

cells. I-K. Conidiophores and conidiogenous cells.

440 ... Silva, Gusmao, & Castafieda-Ruiz

20 pm 20 pm

—

20 um 20 pm

_—__—

Fic. 2. Phaeoschizotrichum ramosum (ex HUEFS 210423). A-B. Hyphae and conidiophore basal

cells. C. Conidiophore and conidiogenous cells. D-I. Branches with fissured outer cell walls and

conidiogenous cells.

Phaeoschizotrichum gen. & sp. nov. & Pyriculariopsis sp. nov. ... 441

CoLonizs on the natural substratum, effuse, hairy, amphigenous, brown.

Mycelium superficial, composed of septate, pale brown, 2-4 um wide, smooth,

branched hyphae, forming an anastomosed network of textura epidermoidea

appearance. CONIDIOPHORES distinct, single, rarely unbranched, mostly

branched above, erect, cylindrical, smooth near the base, ventricose, closely

septate, with fissured outer cell walls near the apex and branches, with up to

10 enteroblastic percurrent extensions in the main axis and branches, brown

below, subhyaline to pale brown above, 100-200 x 5-10 um. CoNIDIOGENOUS

CELLS polyblastic, cylindrical-subulate, slightly denticulate, weakly melanized

at the conidiogenous loci, subhyaline to pale brown, sympodially elongated

after enteroblastic percurrent extensions, smooth sometimes cracked, furrowed

or leprous near the percurrent elongation, 15-50 x 4-5 um. Conrpia solitary,

subulate, narrowly obclavate, truncate, slightly obscure at the base, rounded

or obtuse at the apex, 1-4-septate, mostly 3-septate, pale brown, 25-68 x

2.5-4 um, smooth, rarely verruculose, dry.

Pyriculariopsis bicolorata C.R. Silva, Gusmao & R.F. Castafieda, sp. nov. Fic. 3

MycoBank MB810714

Differs from Pyriculariopsis spp. by smaller, 1-septate, bicolored conidia.

Type: Brazil, Bahia, Piata, Serra da Tromba, 13°07’S 41°50°W, on dead leaf of Calophyllum

brasiliense, 2 X. 2013, coll. C.R. Silva (Holotype: HUEFS 210424).

Erymo_oey: Latin, bicolorata, referring to the two-colored conidia.

CoLonizs on the natural substratum, effuse, hairy, hypophyllous, brown.

Mycelium superficial and immersed, composed of septate, branched, brown,

1-2 um wide, smooth hyphae. ConrpiopHores distinct, single, develop

as lateral branch of hyphae, unbranched, cylindrical, straight to slightly

flexuous, 4—7-septate, brown dark at the base becoming paler to the apex, with

2-5 percurrent extensions, 75-175 x 2.5-4 um, smooth. Conidial secession

schizolytic. CONIDIOGENOUS CELLS polyblastic, integrated, mostly terminal,

sometimes intercalary, sympodially elongated, slightly denticulate, pale

brown, 15-47.5 x 2.5-4 um, slightly cicatrized at the loci. Conrp1 obclavate,

truncate at the base, obtuse and rounded apex, slightly constricted at the septa,

1-septate, bicolored, brown basal cell, subhyaline apical cell, single, dry, 12-18 x

2.9-4 um.

Note: A key to eight Pyriculariopsis species with conidial illustrations was

provided by Iturriaga et al. (2008); subsequently Soares et al. (2011) named

P. calatheae, which is characterized by cylindrical-fusiform to obclavate,

18-36 x 5-7 um, 1-2-septate, often with a rostrate apical cell, 3-12 x 2-4 um,

pale brown to subhyaline. None of the nine previously described species of

Pyriculariopsis is close to or resembles P. bicolorata.

442 ... Silva, Gusmao, & Castafieda-Ruiz

A i C D E

5 yum

20 um

Fic. 3. Pyriculariopsis bicolorata (ex HUEFS 210424). A-E. Conidia. EF Conidium and

conidiogenous cells. G-H. Conidiogenous cells. I. Conidiophore with external hyphae.

Phaeoschizotrichum gen. & sp. nov. & Pyriculariopsis sp. nov. ... 443

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li and Dr. Raghvendra

Singh for their critical review of the manuscript. The authors are grateful to PPBio

Semiarido and Edital Universal do CNPq for financial support (Proc. 558317/2009-0

and 558317/2009-0). The authors thank the Programa de Pés-Graduagao em Biologia

de Fungos (PPGBF/ UFPE). C.R. Silva and L.EP. Gusmao extend their gratitude to

CNPq for financial support (Proc. 132415/2013-5). RFCR is grateful to Cuban Ministry

of Agriculture and “Programa de Salud Animal y Vegetal’, project P131LH003033

for facilities. We acknowledge the assistance provided by Dr. PM. Kirk and

Drs. V. Robert and A. Decock through the IndexFungorum and MycoBank websites.

Dr. Lorelei Norvell’s editorial and Dr. Shaun Pennycook’s nomenclatural reviews are

greatly appreciated.

Literature cited

Castaneda Ruiz RE. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Ellis MB. 1971. Dematiaceous hyphomycetes. Kew, Commonwealth Mycological Institute.

Iturriaga T, Fernandéz R, Castafieda-Ruiz RF, Minter DW, Heredia GA. 2008. A new anamorphic

fungus from Venezuela: Pyriculariopsis formosa. Mycotaxon 105: 337-342.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes.

CBS Biodiversity Series 9. 997 p. http://dx.doi.org/10.3767/003158511X617435

Soares DJ, Rocha FB, Barreto RW. 2011. Pyriculariopsis calatheae sp. nov., a novel anamorphic

hyphomycete from the Atlantic forest of Brazil causing leaf spots on Calathea longifolia. Mycol.

Progress 10: 315-321. http://dx.doi.org/10.1007/s11557-010-0704-3

MY COTAXON

http://dx.doi.org/10.5248/130.445

Volume 130, pp. 445-449 April-June 2015

Anaexserticlava caatingae, a new conidial fungus

from the semi-arid Caatinga biome of Brazil

TASCIANO DOS SANTOS SANTA IZABEL’, DAvI AUGUSTO CARNEIRO DE ALMEIDA’,

JOSIANE SANTANA MONTEIRO’, MARCOS FABIO OLIVEIRA MARQUES’,

Luis FERNANDO PASCHOLATI GUSMAO”*, & RAFAEL FE CASTANEDA-RUIZ?

Universidade Estadual de Feira de Santana, Departamento de Ciéncias Bioldgicas,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Bahia, Brazil

?Universidade do Estado da Bahia, Departamento de Educagao, Campus VII,

BR407 Km 127, 48970-000, Senhor do Bonfim, Bahia, Brazil

*Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—An interesting fungus collected during investigations of microfungi on dead

plant material in a semi-arid region of northeast Brazil is herein described and illustrated.

Anaexserticlava caatingae gen. & sp. nov. is distinguished by differentiated dark brown

conidiophores, holoblastic percurrent elongated conidiogenous cells that produce several

sessile subcylindrical hyaline separating cells, and clavate to ovoid 5-6-distoseptate brown

conidia.

KEY worDs— asexual ascomycete, systematic, leaf litter, tropics

The semi-arid region of Brazil is located almost exclusively in the northeast

region of the country occupying an area of approximately 900,000 km? (Giulietti

et al. 2006). The Caatinga biome is the predominant vegetation and several new

microfungi have recently been described in this region (Almeida et al. 2013,

2014; Barbosa et al. 2013; Cruz et al. 2012; Fiuza et al. 2014; Leado-Ferreira et

al. 2013; Silva & Gusmao 2013; Silva et al. 2014). During a mycological survey

of conidial fungi from the semi-arid region in Bahia and Piaui States, an

interesting fungus was collected and showed remarkable differences from all

previously described hyphomycete genera (Seifert et al. 2011). The specimen is

therefore described as a new genus.

446 ... Santa Izabel & al.

The samples of dead plant material were placed in paper bags, taken to the

laboratory, and prepared according to Castafeda-Ruiz (2005). Mounts were

prepared in PVL (polyvinyl alcohol, lactic acid, and phenol) and measurements

were made at 1000x magnification. Microphotographs were obtained with

an Olympus BX51 microscope equipped with bright field and Nomarski

interference optics. The type specimen is deposited in the Herbarium of

Universidade Estadual de Feira de Santana (HUEFS) and an additional specimen

collected from another field expedition in the Herbarium of Universidade do

Estado da Bahia (HUNEB/SB), Brazil.

Anaexserticlava T.S. Santa Izabel, R.F. Castafieda & Gusmao, gen. nov.

MycoBank MB810632

Differs from Exserticlava by sessile, subcylindrical, hyaline separating cells and

rhexolytic conidial secession.

TYPE SPECIES: Anaexserticlava caatingae T.S. Santa Izabel et al.

ErymMo.oey: Latin, Anaexserticlava, named after its morphological similarities to the

hyphomycete genus Exserticlava.

Asexual fungi. CONIDIOPHORES distinct, single, unbranched, erect, straight,

septate, smooth, dark brown. CoNIDIOGENOUS CELL holoblastic, integrated,

terminal, with several enteroblastic percurrent extensions; conidial secession

rhexolytic. SEPARATING CELLS sessile, cylindrical to subcylindrical, hyaline.

Conipia solitary, clavate to ovoid, distoseptate, pedicellate, smooth, first pale

brown, later brown.

Anaexserticlava caatingae T.S. Santa Izabel, R.E. Castafieda & Gusmao, sp. nov.

MycoBank MB810633 FIG. 1

Differs from Exserticlava spp. by having sessile, subcylindrical to doliiform, hyaline

separating cells and pedicellate conidia.

Type: Brazil, Piaui, Caracol, Serra das Confusées, 9°13’S, 43°29’W, on decaying leaves of

an unidentified plant, 4.V.2011, coll. D.A.C. Almeida (Holotype: HUEFS173319).

ErymMo_oey: Latin, caatingae, referring to the Caatinga biome.

Co.tonigs on the natural substrate effuse, hairy, brown. Mycelium mostly

immersed, composed of branched, septate, smooth, pale brown hyphae,

1-2 um diam. CoNnIDIOPHORES distinct, single, unbranched, erect, straight

or slightly flexuous, septate, smooth, dark brown, pale brown toward the

apex, 100-200 x 7.5-10 um. CONIDIOGENOUS CELLS holoblastic, integrated,

terminal, with several enteroblastic percurrent extensions, smooth, pale brown.

SEPARATING CELLS sessile, doliiform to subcylindrical, hyaline or subhyaline.

Conidial secession rhexolytic. Conip1 solitary, clavate to ovoid, rounded at

the ends, pedicellate, 5-6-distoseptate, smooth, pale brown at first, brown after

maturation, 20-27.5 x 7.5-12.5 um; pedicels 3-4.5 x 1.5-3 um.

Anaexserticlava caatingae gen. & sp. nov. (Brazil) ... 447

Fic. 1. Anaexserticlava caatingae (holotype HUEFS 173319): A-F, Conidiophores, conidiogenous

cells, separating cells, and conidia. G—J. Conidia. Scale bars = 10 um.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, Banta, Morro do Chapéu, on decaying

leaves of an unidentified plant, 22.V.2008, coll. E.B. Santos (HUNEB/SB1484).

448 ... Santa Izabel & al.

Fic. 2. Exserticlava vasiformis (ex HUEFS 155041): A-C. Conidiophores, conidiogenous cells, and

conidia. Scale bars = 10 um.

Note: Among the hyphomycete genera compiled by Seifert et al. (2011), only

Exserticlava S. Hughes (Hughes 1978) has some similarities to Anaexserticlava,

Anaexserticlava caatingae gen. & sp. nov. (Brazil) ... 449

but Exserticlava vasiformis (Matsush.) S$. Hughes has schizolytic conidial

secession and the conidia are not pedicellate at the base (Fic. 2).

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li (The Connecticut

Agricultural Experiment Station Valley Laboratory, USA) and Dr. Xiu Guo Zhang

(Department of Plant Pathology, Shandong Agricultural University, Taian, China) for

their critical review of the manuscript. The authors thank the Program of Research on

Biodiversity in the Brazilian Semi-arid (CNPq Proc. 558317/2009-0) and “Programa

Ciéncia sem Fronteiras” RFCR is also grateful to the Cuban Ministry of Agriculture

for supporting and facilities through “Programa de Salud Animal y Vegetal’, project

P131LH003033. We acknowledge the assistance provided by Dr. P.M. Kirk and Drs.

V. Robert and A. Decock through the IndexFungorum and Mycobank websites. Dr.

Lorelei Norvell’s editorial and Dr. Shaun Pennycook’s nomenclatural reviews are greatly

appreciated.

Literature cited

Almeida DAC, Cruz ACR, Marques MFO, Gusmao LFP. 2013. Conidial fungi from semi-arid

Caatinga biome of Brazil. New and interesting Zanclospora species. Mycosphere 4: 684-692.

http://dx.doi.org/10.5943/mycosphere/4/4/4

Almeida DAC, Miller AN, Gusmao LFP. 2014. New species and combinations of conidial

fungi from the semi-arid Caatinga biome of Brazil. Nova Hedwigia 98: 431-447.

http://dx.doi.org/10.1127/0029-5035/2013/0162

Barbosa FR, Raja HA, Shearer CA, Gusmao LFP. 2013. Some freshwater fungi from the Brazilian

semi-arid region, including two new species of hyphomycetes. Cryptogamie Mycologie 34:

243-258. http://dx.doi.org/10.7872/crym.v34.iss2.2013.243

Castanieda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Cruz ACR, Gusmao LFP, Castafeda-Ruiz RE, Stadler M, Minter DW. 2012. Zelodactylaria, an

interesting new genus from semi-arid northeast Brazil. Mycotaxon 119: 241-248.

http://dx.doi.org/10.5248/119.241

Fiuza PO, Gusmao LFP, Cruz ACR, Castafieda-Ruiz RE 2014. Conidial fungi from the semiarid

Caatinga biome of Brazil: a new species of Pseudoacrodictys. Mycotaxon 127: 33-37.

http://dx.doi.org/10.5248/127.33

Giulietti AM, Harley RM, Queiroz LP, Rapini A. 2006. To set the scene. 15-19, in: AM Giulietti

et al. (eds). Towards greater knowledge of the Brazilian semi-arid biodiversity. Ministério da

Ciéncia e Tecnologia. Brasilia.

Hughes SJ. 1978. New Zealand Fungi. 25. Miscellaneous species. New Zealand Journal of Botany

16: 311-370. http://dx.doi.org/10.1080/0028825X.1978.10425143

Leao-Ferreira SM, Gusmao LFP, Castafieda-Ruiz RE 2013. Conidial fungi from the semi-arid

Caatinga biome of Brazil. Three new species and new records. Nova Hedwigia 96: 479-494.

http://dx.doi.org/10.1127/0029-5035/2013/0084

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p. http://dx.doi.org/10.3767/003158511X617435

Silva SS, Gusmao LFP. 2013. Conidial fungi from the semi-arid Caatinga biome of Brazil. A new

species of Dictyochaeta. Mycosphere 4: 701-705. http://dx.doi.org/10.5943/mycosphere/4/4/6

Silva SS, Cruz ACR, Gusmao LFP, Castafeda-Ruiz RF. 2014. Diplococcium variegatum, a new

conidial fungus from the semi-arid Caatinga biome of Brazil. Mycotaxon 127: 59-62.

http://dx.doi.org/10.5248/127.59

MY COTAXON

http://dx.doi.org/10.5248/130.451

Volume 130, pp. 451-454 April-June 2015

A new species of Endophragmiella from Guizhou, China

YING-RuI Ma, JI-WEN XIA, & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University, Taian, 271018, China

*CORRESPONDENCE TO: zhxg@sdau.edu.cn, sdau613@163.com

ABSTRACT — Endophragmiella selenosporellaria sp. nov. was collected on dead branches of

an unidentified broadleaf tree from southern China and is characterized by its branched

conidiophores and Selenosporella-like synanamorph developed from the apical cell of the

conidium. It is described, illustrated, and compared with similar species. The specimens are

deposited in Herbarium of Shandong Agricultural University, Plant Pathology (HSAUP) and

Mycological Herbarium, Institute of Microbiology, Chinese Academy of Sciences (HMAS).

KEY worDs — anamorphic fungi, taxonomy

Introduction

Endophragmiella is a genus established by Sutton (1973) with two species,

E. pallescens B. Sutton (the type species) and E. canadensis (Ellis & Everh.)

B. Sutton. Hughes (1979) later emended the genus and gave a very detailed

account of conidiogenesis and generic concepts, describing the genus as

comprising hyperparasites with often frequently branching conidiophores

and conidia that bear a distinct frill. Endophragmiella is mainly characterized

by solitary acrogenous conidia that secede rhexolytically from monoblastic,

integrated, terminal, and determinate or percurrently extending conidiogenous

cells (Sutton 1973, Ellis 1976, Hughes 1979, Wu & Zhuang 2005). At present,

over 90 species are accepted in the genus (MycoBank 2014, Ma et al. 2011, Ren

et al. 2011). The taxonomic classification of Endophragmiella species is based

primarily on morphological characteristics including conidial shape, size,

pigmentation, septation, and presence or absence of a rostrum at the apex and

the frill at the base.

During an investigation of saprobic microfungi on dead branches of

Southern China, an interesting fungus with morphological features typical

of Endophragmiella (Sutton 1973) was collected. It differs significantly from

452 ... Ma, Xia, & Zhang

previously described Endophragmiella species and is proposed here as new to

science.

Materials and methods

Samples of decaying wood were collected from subtropical forests of Guizhou

province, China, placed in separate zip-lock plastic bags, taken to the laboratory, and

then incubated at 27°C for more than 2 weeks in an artificial climate box in 9 cm

diameter plastic Petri dishes containing moistened filter paper. Samples were examined

under an Olympus SZ61 dissection microscope. All microscopic characteristics were

determined on the basis of measurements of 50 mature conidia and 30 conidiophores

mounted in lactophenol at 60x and 100x magnification, and photographed with an

Olympus BX51 microscope.

Endophragmiella selenosporellaria Y.R. Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 812602

Differs from Endophragmiella corticola by its slightly smaller conidia with a subconical

apical elongation and its Selenosporella-like synanamorph and from E. verticillata by its

irregularly branched conidiophores.

Type: China, Guizhou Province: Mount Leigong, on dead branches of an unidentified

broadleaf tree, 9 Sep. 2013, Y.R. Ma (Holotype, HSAUP H4320; isotype, HMAS 243457).

EryMo_oey: refers to the synanamorphic Selenosporella-like conidia.

Co onigs on the natural substrate effuse, brown. Mycelium superficial and immersed,

composed of branched, septate, pale brown to brown, smooth-walled hyphae.

CONIDIOPHORES macronematous, singly or in small groups, straight or flexuous,

multiseptate, smooth, light brown or yellowish, paler towards the apex, determinate

without percurrent proliferations, 48.5-117.5 x 3.0-4.0 um. Branches arising more or

less 45° to 50° from the main stalk, 8.5-32.5 um long, 0-2-septate. CONIDIOGENOUS

CELLS tapered at the apex, monoblastic, terminal, integrated, slightly swollen, pale brown,

subhyaline to hyaline. Conidial secession rhexolytic. Conrp1A solitary, acrogenous,

elongated ellipsoid to obclavate, basal cell brown to dark brown, often with a small basal

frill, apical cell subconical elongated, subhyaline to hyaline, (0-)2-3-septate, slightly

constricted at septa, 14.5-22.0 x 4.5-6.5 um. SYNANAMORPH Selenosporella-like conidia

filiform, acerose, hyaline, aseptate, 8-12 x 1.0-1.5 um, developed from apical cell of the

conidia.

ComMENts - Endophragmiella selenosporellaria possesses conidia most

similar in shape to those of E. corticola, E. verticillata, E. acuta, E. curvata, and

E. cesatii. However, E. corticola differs by its slightly bigger (16.5-25 x 5.5-7.5

um) light-colored conidia without a subconical apical elongation and its lack of

a Selenosporella-like synanamorph (Kirk 1982); E. curvata has predominantly

2-septate conidia and unbranched conidiophores (Hughes 1979); E. cesatii has

wider (11-12.5 um), predominantly 3-septate conidia with subhyaline basal

cell and unbranched conidiophores (Hughes 1979); E. acuta has wider (8-10

um), rostrate, predominantly 3-septate conidia (Wu & Zhuang 2005); and

Endophragmiella selenosporellaria sp. nov. (China) ... 453

99 ee

Fic. 1. Endophragmiella selenosporellaria (holotype, HSAUP H4320).

a. Conidiophores with conidia. b. Conidiogenous cells and conidia. c. Conidia

E. verticillata has shorter (12.5-16.2 um), 3-septate (not constricted) conidia

and verticillately branched conidiophores (Hughes 1978).

Acknowledgments

The authors express gratitude to Dr. R.E Castafieda-Ruiz and Dr. De-Wei Li for

serving as pre-submission reviewers and for their valuable comments and suggestions.

This project was supported by the National Natural Science Foundation of China (Nos.

31093440, 31230001) and the Ministry of Science and Technology of the People’s

Republic of China (Nos. 2006FY120100).

Literature cited

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Surrey, England.

Hughes SJ. 1978. Endophragmiella verticillata. Fungi Canadenses, No. 130.

Hughes SJ. 1979. Relocation of species of Endophragmia auct. with notes on relevant generic names.

New Zealand J. Bot. 17: 139-188.http://dx.doi.org/10.1080/0028825X.1979.10426887

454 ... Ma, Xia, & Zhang

Kirk PM. 1982. New or interesting microfungi IV. Dematiaceous hyphomycetes from Devon.

Trans. Br. Mycol. Soc. 78: 55-74. http://dx.doi.org/10.1016/S0007-1536(82)80077-6

Mycobank. 2014. MycoSearch database search. International Mycological Association, [Accessed:

14 November 2014]. http://www.mycobank.org/Biolomics.aspx?

Ma LG, Ma J, Zhang YD, Zhang XG. 2011. Taxonomic studies of Endophragmiella from southern

China. Mycotaxon 117: 279-285. http://dx.doi.org/10.5248/117.279

Ren SC, Ma J, Zhang XG. 2011. A new species and new records of Endophragmiella from China.

Mycotaxon 117: 123-130. http://dx.doi.org/10.5248/117.123

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycol. Pap. 132.143 p.

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China.

Fungal Divers. Res. Ser. 15. 351 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.455

Volume 130, pp. 455-464 April-June 2015

Peziza succosella and its ectomycorrhiza associated with

Cedrus deodara from Himalayan moist temperate forests

of Pakistan

SANA JABEEN , TAYIBA ASHRAF, & ABDUL NASIR KHALID

Department of Botany, University of the Punjab,

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

" CORRESPONDENCE TO: ectomycorrhizae@gmail.com

ABSTRACT —Ascomata of Peziza succosella and its ectomycorrhiza associated with Cedrus

deodara were collected from the Himalayan moist temperate forests of Pakistan. The fungal

partner in the ectomycorrhiza was identified by sequencing the rDNA internal transcribed

spacer region. The morphology of the taxon is also described. Ascomata are characterized

by grayish brown deep cupulate apothecia while the ectomycorrhiza is characterized by a

thin pseudoparenchymatous mantle with a few emanating elements. Peziza succosella and

its ectomycorrhiza are an addition to the mycoflora of Pakistan and this is the first report of

ectomycorrhizal association of P. succosella with Himalayan cedar.

Key worps — Cedrus deodara, conifers, ITS, Pezizaceae, morphotype

Introduction

Peziza Dill. ex Fr. is the largest genus within Pezizaceae, with approximately

100 accepted species (Kirk et al. 2008) distributed worldwide. Ten Peziza

species have been reported from Pakistan: P badiofusca (Boud.) Dennis,

P. cerea Sowerby, P. gerardii Cooke, P. michelii (Boud.) Dennis, P. micropus Pers.,

P. pakistanica (S. Ahmad) S. Ahmad, P repanda Wahlenb., P succosa Berk.,

P. vesiculosa Bull., and P. violacea Pers. (Ahmad et al. 1997, Ashraf & Khalid

2012, Ashraf et al. 2012).

The genus is characterized by epigeous cupulate to discoid sessile to subsessile

fleshy ascomata ranging from a few millimetres to more than 10 centimetres

in diameter. The majority of the species are considered to be saprotrophs,

although ectomycorrhizal species have also been reported (Maia et al. 1996,

Hansen et al. 2001, Tedersoo et al. 2006, Ashraf et al. 2012, Jabeen et al. 2012,

Jabeen & Khalid 2014). Generally ectomycorrhizae (EcM) of Pezizales possess

456 ... Jabeen, Ashraf, & Khalid

a thin pseudoparenchymatous mantle, infrequent radiating elements, and no

clamp connections (Agerer 1991a, 2001, 2006). Rhizomorphs, if present, do not

connect to the ascomata. Identification of EcM of these fungi is difficult using

morphological and anatomical methods, but molecular methods have proved

to be an efficient identification tool. Combining morphological-anatomical

characters with molecular markers is helpful for precise identification of these

fungi (Tedersoo et al. 2006, Ashraf & Khalid 2012, Ashraf et al. 2012).

Some pezizalean EcM have already been reported from Pakistan, and most

are known to associate with deciduous trees (Ashraf et al. 2012, Jabeen et al.

2012, Jabeen & Khalid 2014). Peziza succosella has been reported previously

from Israel, Denmark, and Italy (Barseghyan & Wasser 2011, Tedersoo et al.

2006, Osmundson et al. 2013). In Pakistan, the species has been collected from

Himalayan moist temperate forests, which extend from Murree hills and Ayubia

to Miandam at 1890 to 2500 m above sea level (a.s.l.) between subtropical pine

forests and sub-alpine forests dominated by Abies pindrow (Royle ex D. Don)

Royle, Cedrus deodara (Roxb. ex D. Don) G. Don, and Pinus wallichiana A.B.

Jacks. along with some deciduous trees, notably species of Populus, Quercus,

and Salix. Cedrus deodara (Himalayan cedar) is an important phytobiont for

many EcM fungi (Wang & Qiu 2006, Vaario et al. 2006, Hibbett & Matheny

2009, Hanif et al. 2012). The present work is the first record of the association of

P. succosella with Himalayan cedar and a new record of this fungus for Pakistan.

Materials & methods

Sampling site

Sampling was carried out during monsoon season from 2010 to 2013. Two sampling

sites in Khyber Pakhtunkhwa (KPK) were selected, Khanspur, Ayubia and Sharan,

Kaghan Valley. These sites are dominated by moist temperate conifer forests. The

areas range from mountains with rugged valleys to undulating and dissected sub-

mountain plateaus and flat mountains at 3000-4000 m a.s.l. (Khan 1999). Mean annual

temperature of these sites is 10°C (Malik & Sukhera 2012), with temperatures down to

3°C curing December and January and up to 26°C during summer; mean annual rainfall

is 1200 mm with 57% humidity (Ahmed et al. 2006). Soil is silt loam to silt clay and non-

calcareous to slightly calcareous (Khan 2004). The pH is slightly acidic (<7) due to high

organic content (3-4%) (Irshad & Khan 2012). Among the conifers Cedrus, Picea, Pinus,

and Taxus dominate, along with some deciduous tree species (Sheikh 1993).

Collection and characterization of ascomata

Specimens were photographed using Nikon D70S digital camera in the field and

were carefully removed. At least three individual fruit bodies were collected from each

sampling site. Morphological features of fresh specimen were recorded, colors were

designated following the Munsell Soil Color Charts (1975), and then the specimens

were dried under fan heater. Specimens were sectioned and mounted on slides, and

anatomical features were examined a microscope (MX4300H, Meiji Techno Co., Ltd.,

Peziza succosella ectomycorrhiza in Pakistan ... 457

Japan). Structural components of the excipulum were studied in free hand sections and

hymenial elements were studied by tearing apart a piece of hymenium using a needle.

Measurements were recorded using Carl Zeiss Jena ocular micrometer and line drawing

were made using Leitz Wetzlar Camera Lucida. The abbreviation n/m/p means n

ascospores were measured from m ascomata and p collections. Voucher specimens were

deposited in the Herbarium, Botany Department, University of the Punjab, Lahore,

Pakistan (LAH).

Collection, isolation, and characterization of EcM

Sampling was carried out very carefully to make sure that the fine roots in the

soil block belonged to the same tree. A 15-cm? soil block was dug with a shovel a few

centimetres from the tree trunk. Five soil cores were taken from each sampling site.

In the laboratory, each soil core was soaked in water for a few hours to loosen the

soil particles and then put on a 2 mm sieve under running water to separate the roots

from the soil. The EcM were carefully sorted into morphotypes under incandescent

light and then under a stereomicroscope (EMZ-5TR, Meiji Techno Co., Ltd., Japan).

The morphotypes were cleaned under the stereomicroscope using a fine brush.

Morphologically identical morphotypes were kept in McCartney bottles in distilled

water for future analysis. Replicates of these morphotypes were kept at 8°C in Eppendorf

tubes containing 2% CTAB buffer for molecular analysis. Morphological characters

(e.g., color, size, ramification, presence or absence of radiating elements) were noted

under the stereomicroscope. Mantle layers (inner and outer) and radiating hyphae were

mounted in trypan blue stain (Agerer 1991b). Hyphae were measured using an ocular

micrometer and drawn using a camera lucida. A voucher specimen was deposited in

LAH Herbarium.

DNA extraction, amplification and sequencing

For molecular analysis, up to 2 mg of ascomatal tissue was placed in one Eppendorf

tube and 2-4 ectomycorrhizal root tips were placed in a separate Eppendorf tube. DNA

was extracted using modified CTAB method (Bruns 1995). Internal Transcribed Spacer

(ITS) regions of nuclear ribosomal DNA (nrDNA) were amplified using universal primer

pair (ITSLF: 5’-cTTGGTCATTTAGAGGAAGT-3’ and ITS4: 5’- TCCTCCGCTTATTGATATGC-3’)

following Gardes & Bruns (1993). The products were sent to Macrogen Inc. (Korea) for

sequencing.

Molecular phylogenetic analysis

Consensus sequences were generated from the obtained sequences and then BLAST

searched at NCBI (http://www.ncbi.nlm.nih.gov/). Sequences with the closest matches

were selected from GenBank to reconstruct a phylogeny. Sequences that showed less

query cover and negative E value were not included. Published sequences of the closest

relatives of the species were also included in the final data set, and Sarcosphaera coronaria

(Jacq.) J. Schrét. was chosen as outgroup (Tedersoo et al. 2006). ClustalW was used to

align sequences in BioEdit software. The sequences were trimmed with the conserved

motifs 5 -(...GAT) CATTA... and...GACCT (CAAA...)-3, and the alignment portions

between them was used to reconstruct phylogeny. Maximum likelihood (ML) analysis

was performed using Jukes-Cantor model in MEGA6 software to test the phylogeny at

1000 bootstraps. Percentage identity and divergence in nrDNA-ITS was analysed using

458 ... Jabeen, Ashraf, & Khalid

MegAlign (DNAStar). Sequences generated in this study were submitted to GenBank

(KM199728, KM199729).

Taxonomy

Peziza succosella (Le Gal & Romagn.) M.M. Moser ex Aviz.-Hersh. & Nemlich,

Israel J. Bot. 23(3): 156 (1974). Pig. 1

APOTHECIA deeply cupulate, sessile to sub-sessile, circular, centrally

attached, initially circular, irregular at older stage, medium-sized (1.5-2.5 cm).

Hymenium smooth, olivaceous grey (10YR6/2), outer surface smooth, pale

grey (5Y7/2). Asci cylindrical, operculate, unitunicate, 8-spored, uniseriate,

strongly amyloid at apex, slightly narrowing towards base, 253-290 x 15-17

um. AscosporEs [50/4/2] ellipsoid, uniguttulate, light brown, ornamented,

(15-)16-18 x 8-11 um (16.93 + 0.815 x 9.66 + 0.59; Qm = 1.75 + 0.12),

ornamentation longitudinal warts to partially reticulate, 1.2-2.0 um high warts.

PARAPHYSES Slender, septate near base, same diam. throughout length, same

length as asci, <4.3-5.0 um wide. ExcIPULUM ectal textura globosa to textura

angularis, slightly brown cells, 13-23 x 15-20 um, globose cells of ectal smaller

as compared to inner angular cells, ectal cells ending in hyphoid extensions that

appear septate, hyaline, blunt ended, 8-11 um wide.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, Ayubia, Khanspur

Village, 2575 m a.s.l, in Himalayan moist temperate forests, on ground, damp soil,

21 August 2010, Tayiba Ashraf; TA-135 (LAH210810); Kaghan Valley, Sharan, 2011

m a.s.l., in Himalayan moist temperate forests, 14 August 2011, Tayiba Ashraf, TA-

221(LAH140811; GenBank KM199729).

Morphological characterization of ECM FIG. 2

ECTOMYCORRHIZAL SYSTEM dichotomous to coralloid, explored at upper

layer of soil, <4 mm long, axis <400 um in diameter, un-ramified ends straight,

not inflated and cylindrical, <400 um in diameter, width of tip base and apex

300 um, grayish brown to dark brown or black at maturity. MANTLE grayish

(5Y7/2) to transparent, distinct, smooth surface, luster matte; host tissue

visible under the mantle surface. RHIZOMORPHS absent. EMANATING HYPHAE

transparent, infrequent, sometimes frequent at few tips, straight, cylindrical

rarely branched.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, Ayubia, Khanspur

Helipad, 2575 m a.s.l., in Himalayan moist temperate forests, mycorrhiza associated

with Cedrus deodara, 21 September 2013, Sana Jabeen; SA116; HP-5 (LAH-EM1-2013;

GenBank KM199728).

Anatomical characteristics of mantle in plan view

Mantle pseudoparenchymatous in all layers, non-gelatinous, angular cells

with blunt corners. OUTER MANTLE LAYER pseudoparenchymatous, cells

angular, (mantle type K, Agerer 1987-2002; Agerer & Rambold 1998), cells

Peziza succosella ectomycorrhiza in Pakistan ... 459

Fic. 1. Peziza succosella (LAH140811). A. Apothecia. B. Asci with ascospores and paraphyses.

C. Ornamented ascospores. D. Part of excipulum. Scale bars: A = 1.5 cm; B, D = 25 um; C = 5 um.

460 ... Jabeen, Ashraf, & Khalid

membranaceously and plasmatically transparent, 11.56 x 10.01 + 9.05 um,

matrix clear. INNER MANTLE LAYER pseudoparenchymatous, angular cells with

blunt corners, (mantle type K, Agerer 1987-2002; Agerer & Rambold 1998),

cells membranaceously and plasmatically transparent, 12.46 x10.05 + 10.07

uum, cell contents clear.

Anatomical characteristics of emanating elements

EMANATING HYPHAE elongated, straight, cylindrical, up to 4.74 um in

diameter, wall up to 0.70 um thick, surface smooth, hyphae septate, septa

frequent, clamps absent, septa 32.70 um apart, contents clear, ramification

rather frequent, Y-shaped, anastomosing; cystidia not observed. RHIZOMORPHS

absent.

Molecular Phylogenetic Characterization

Sequences of the ITS region of nrDNA of fruit body and EcM were BLAST

searched at NCBI. BLAST search revealed that the subject sequences were 99%

identical with P. succosella (JF908535) andaspecimen identified as P. subumbrina

Boud. (JF908552) from Italy with 100% and 96% query cover respectively, and

0.0 E value. Both Pakistani sequences — TA-221 (fruit body) and SA116 (EcM)

— clustered with P succosella from Italy and Denmark (DQ200841) with 99%

boot strap support; these form a sister clade with P infossa and P. succosa in

the phylogenetic tree with 92% bootstrap support (Fic. 3). Both sequences

showed a 99.8% genetic identity with P. succosella (JF908535), a 97.2% with

the questionably identified Italian specimen of P. subumbrina (JF908552), and

a 92.7 % genetic identity with P. succosella (DQ200841) from Denmark with

0 value of genetic divergence. The low value of genetic identity of P. succosella

(DQ200841) is due to low query cover (89%) in BLAST.

Discussion

Pezizalean species constitute an important part of mycobiont diversity

in EcM fungal communities (Tedersoo et al. 2006). Many EcM fungal taxa

belonging to this group have been reported from Himalayan moist temperate

forests of Pakistan (Ashraf et al. 2012, Hanif et al. 2012). ECM morphology

of pezizalean taxa is little studied. Previously published morphological and

anatomical descriptions of pezizalean EcM cover only a few genera. EcM of

P. succosella are grayish brown and showed dichotomous to coralloid type of

ramification, presence of a pseudoparenchymatous thin mantle layer with few

emanating hyphae, and lack of clamped septa. These characters are distinct

from the previously reported EcM of P. michelii having yellow green to olive

green morphotypes (Tedersoo et al. 2006) and cream white with brown to

black older tips (Ashraf et al. 2012). Cells of the mantle layer of P. michelii

differ greatly in size and shape from each other while the cells of mantle layer

of P. succosella showed comparatively less variability in size and shape among

Peziza succosella ectomycorrhiza in Pakistan ... 461

Fic. 2. EcM of Peziza succosella. A. Ectomycorrhizal system. B. Root tip morphology. C. Inner

mantle layer. D. Outer mantle layer. E. Emanating hyphae. Scale bars: A = 0.5 mm; B = 0.25 mm;

C-E= 10 um.

themselves. Sequences of the fruit body and EcM along with morphological

descriptions were found helpful in identification. ITS phylogenetic analysis

revealed that the sequences derived from ascocarp and ectomycorrhizal tissue

462 ... Jabeen, Ashraf, & Khalid

100 p— AF491544 Peziza varia

71 AF491545 Peziza varia

AF491574 Peziza echinispora

100 AF491575 Peziza echinispora

AF491581 Peziza arvernensis

39 99 AF491583 Peziza arvemensis

100 - AF491612 Peziza alcis

AF491611 Peziza alcis

100 p- AF 491620 Peziza nivalis

57 ih AF491619 Peziza nivalis

95 AF491593 Peziza fimeti

96 AF491594 Peziza fimeti

100 ; AF491623 Peziza vesiculosa

AF491624 Peziza vesiculosa

90 AF491622 Peziza ammophila

84 94 AF491621 Peziza ammophila

AF491628 Peziza subcitrina

100 !'AF491627 Peziza subcitrina

JN002180 Peziza ostracoderma

100 + AY818334 Peziza ostracoderma

DQ200839 Peziza michelii

100 ' JF908562 Peziza michelii

a JF908556 Peziza succosa

JF908556 Peziza succosa

DQ200840 Peziza succosa

99 y DQ974817 Peziza infossa

92) L AY830853 Peziza infossa

JF908552 Peziza succosella (as "P subumbrina")

60 | |@ KM199729_Peziza succosella

@ KM199728_Peziza succosella

JF908535 Peziza succosella

DQ200841 Peziza succosella

AF 133172 Sarcosphaera coronaria _]Out group

Fic. 3. Molecular phylogenetic analysis of Peziza succosella and related species. New sequences

generated from Pakistan are marked with @. Genbank accession numbers of all taxa are given.

The percentage of trees in which the associated taxa clustered together at 1000 bootstraps is shown

next to the branches. The tree is drawn to scale, with branch lengths measured in the number of

substitutions per site. The analysis involved 33 nucleotide sequences. There were a total of 885

positions in the final dataset.

clustered with P. succosella from Italy and Denmark with very strong bootstrap

value. Its ectomycorrhiza has previously been undescribed. The molecular

distinction among specimens from Pakistan and other countries is minimal,

placing the ascocarps and EcM of Peziza succosella in the same species, despite

the large geographic distance. This indicated a wide species distribution and a

circum-Mediterranean distribution pattern.

Acknowledgments

Sincere thanks to Dr. Tine Grebenc (Slovenian Forestry Institute, Ljubljana) for

his help in improving the manuscript. We are also thankful to Dr. T.-K. Arun Kumar

Peziza succosella ectomycorrhiza in Pakistan ... 463

(The Zamorin’s Guruvayurappan College, India) and Prof. Donald H. Pfister (Harvard

University Herbaria, Cambridge, MA, USA) for acting as pre submission reviewers.

Thanks to Mr. Hassan Ayub for his help in collecting EcM. The work is financially

supported by Higher Education Commission (HEC)-Pakistan under Indigenous PhD

Fellowships for 5000 Scholars, HEC (Phase-II).

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MY COTAXON

http://dx.doi.org/10.5248/130.465

Volume 130, pp. 465-469 April-June 2015

Cryptocoryneum parvulum, a new species

on Araucaria angustifolia (Brazilian pine)

SILVANA SANTOS DA SILVA', LUIS FERNANDO PASCHOLATI GUSMAO™,

& RAFAEL F. CASTANEDA-RUIZ?

‘Departamento de Ciéncias Biologicas, Laboratorio de Micologia, Universidade Estadual

de Feira de Santana, BR116 KM 03, 44031-460, Feira de Santana, Brazil

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—Cryptocoryneum parvulum sp. nov., collected on decaying needle-like leaves

of Araucaria angustifolia, is described and illustrated. It is distinguished by clavate inflated

conidiophores and small cheiroid conidia with black cap cells and four pendulous arms of

pale brown cells. Two other Cryptocoryneum species, C. condensatum and C. rilstonei, are

newly recorded and illustrated. A key to Cryptocoryneum species found in Brazil is provided.

KEY worDSs— asexual fungi, systematics, subtropical fungi

Introduction

During a mycological survey of fungi associated with litter at a Brazilian

araucaria forest, an interesting Cryptocoryneum specimen was collected

from decaying needle-like leaves. It showed remarkable differences from all

previously described Cryptocoryneum species (Ellis 1971, 1976) and is therefore

described here as new.

Materials & methods

Samples of litter were placed in paper bags, taken to the laboratory, and prepared

according to Castafieda-Ruiz (2005). Mounts were prepared in PVL (polyvinyl alcohol,

lactic acid, and phenol) and measurements were made at a magnification of x1000.

Micrographs were obtained with an Olympus BX51 microscope equipped with bright

field and Nomarski interference optics. The type specimen is deposited in the Herbarium

of Universidade Estadual de Feira de Santana (HUEFS).

466 ... Silva, Gusmao, & Castafieda

Taxonomy

Cryptocoryneum parvulum S.S. Silva, Gusmao & R.F. Castafieda, sp. nov. Fig.1

IF 550828

Differs from other Cryptocoryneum spp. by its inflated conidiophores and conidiogenous

cells and its smaller, cheiroid conidia with dark cap cells and four pendulous, 3-4-septate,

pale brown to subhyaline arms.

Type: Brazil, Rio Grande do Sul State: SAo Francisco de Paula, Floresta Nacional de

Sao Francisco de Paula, 29°25 § 50°23 W, 838 m alt., on decaying needle-like leaves

of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae), 27.V1II.2014, coll. S.S. Silva.

(Holotype, HUEFS 210439).

Erymo oey: Latin, parvulum, referring to the small conidia.

CoNnIDIOMATA sporodochial, pulvinate, 75-140 um diam., scattered, black.

Mycelium superficial and immersed. Hyphae septate, branched, 2.5-3 um

diam., smooth, brown. Conip1iopHoRes distinct, clavate, inflated, single,

erect, straight, close packet, pale brown, 0-2-septate, smooth, 15-25 x 9X16.5

um. CONIDIOGENOUS CELLS monoblastic, clavate to cylindrical or doliiform,

integrated, terminal, determinate. Conip1A solitary, acrogenous, cheiroid,

branched, 12-23 x 10.5-15 um, with black cap cells and four pendulous pale

brown arms, smooth, 3-4-septate, 2.5-5 um wide.

Note: Cryptocoryneum Fuckel (Ellis 1971, 1976, Schoknecht & Crane 1977)

is characterized by monoblastic conidiogenous cells, and cheiroid conidia

composed of a series of dark brown cap cells from which the pendulous arms

develop.

Additional Cryptocoryneum records from Brazil

Cryptocoryneum condensatum (Wallr.) E.W. Mason & S. Hughes ex S. Hughes,

Can. J. Bot. 36: 758 (1958). Fic. 2 A-D

SPECIMEN EXAMINED: BRAZIL, R10 GRANDE DO SUL STATE, Sao Francisco de Paula,

Floresta Nacional de Sao Francisco de Paula, 29°25 S 50°23 W, 838 m alt., on decaying

needle-like leaves of Araucaria angustifolia, 29.1X.2014, coll. $.S. Silva (HUEFS 210440).

Norte: A new record for Brazil.

Cryptocoryneum rilstonei M.B. Ellis, Mycol. Pap. 131: 2 (1972). Fic. 2 E-H

SPECIMEN EXAMINED: BRAZIL, R10 GRANDE DO SUL STATE, Sao Francisco de Paula,

Floresta Nacional de Sao Francisco de Paula 29°25 S 50°23 W, 838 m alt., on decaying

needle-like leaves of Araucaria angustifolia, 21.VIII.2014, coll. S.S. Silva (HUEFS

210441).

Norte: A new record for Brazil.

Cryptocoryneum parvulum sp. nov. (Brazil) ...

20m

20)Lm

Fic. 1. Cryptocoryneum parvulum (holotype, HUEFS 210439): A-B. Sporodochia on natural

substrate. C-E. Sporodochia. FE Conidiophores, conidiogenous cells and conidia. G-H.

Conidiogenous cell and conidia. I. Conidia.

468 ... Silva, Gusmao, & Castafieda

20um

Fic 2. Cryptocoryneum condensatum (HUEFS 210440): A. Sporodochium on the natural

substratum. B-D. Conidia. Cryptocoryneum rilstonei (HUEFS 210441): E. Sporodochium on the

natural substratum. FE. Sporodochium. G-H. Conidia.

Cryptocoryneum parvulum sp. nov. (Brazil) ... 469

Key to Cryptocoryneum species found in Brazil

LeGanidrophores:eylindpical Whi, 6 hen beh n Seth Rhee ao hae hg LaBrie ea, Sn core, Caaf 2

1. Conidiophores clavate, inflated, 15-25 um long; conidia 12-23 x 10,5-15 um,

with dark cap cells and 4 pendulous pale brown arms, 3—-4-septate,

2 DES UE WIME corey Ghee owe dors beh de Fh et F Kigitoye # MG teat BMG HAG ee ES C. parvulum

2. Conidiophores <90 tum long; conidia 40-85 x 20-35 um,

with 6-14 pendulous pale brown to yellowish brown or subhyaline arms,

15-20-septate, 2.5-4.5 um wide .............. eee eee ee eee ee C. condensatum

2. Conidiophores <45 um long; conidia 20-45 x 12-30 um,

with 3-9 pendulous subhyaline or pale brown arms,

BOSE Pate AR 6 PTA! oslo es cls reals wets eats Heals COEli yoo C. rilstonei

Acknowledgments

The authors express their sincere gratitude to Prof. Bryce Kendrick and Dr. De-Wei

Li for their critical review of the manuscript. The authors are grateful to Dr. P. M. Kirk

for his taxonomic commentaries and opinion. The authors are grateful to the CNPq

(proc. 141475/2013-7) for financial support and the “Programa de Pdos-graduacao em

Botanica/UEFS” and ICMBIO for permission to collected microfungi in the “Floresta

Nacional de Sao Francisco de Paula” (proc. 42334-1). The authors acknowledge

the support provided by “Programa Ciéncia sem Fronteiras’. RFCR is grateful to the

Cuban Ministry of Agriculture and “Programa de Salud Animal y Vegetal” (project

P131LH003033) for facilities. We acknowledge the assistance provided by Dr. P.M.

Kirk and Drs. V. Robert and A. Decock through the Index Fungorum and MycoBank

websites. Dr. Lorelei Norvell’s editorial and Dr. Shaun Pennycook’s nomenclatural

reviews are greatly appreciated.

Literature cited

Castanieda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey.

http://dx.doi.org/10.1016/S0181-1584(01)01057-0

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Surrey. http://dx.doi.org/10.5248/109.221

Schoknecht JD, Crane JL. 1977. Revision of Torula and Hormiscium species. Torula occulta,

T. diversa, T. elasticae, T. bigemina and Hormiscium condensatum reexamined. Mycologia 69:

533-546. http://dx.doi.org/10.2307/3758557

MY COTAXON

http://dx.doi.org/10.5248/130.471

Volume 130, pp. 471-478 April-June 2015

Two new species and

a new record of Leptogium from China

Hua-Jrz Liu*, MAN-QING XI,

JIAN-SEN Hu, & QING-FENG Wu

Key Lab of Microbial Diversity Research and Application of Hebei Province,

College of Life Sciences, Hebei University, Baoding, 071002, China

* CORRESPONDENCE TO: liuhuajie@foxmail.com

ABSTRACT — Leptogium taibaiense sp. nov. and L. wangii sp. nov. both have short hairs

composed of cylindrical cells on their lower surfaces and are glabrous and smooth on their

upper surfaces. Leptogium burgessii is a new record for China.

KEY worps —taxonomy, Ascomycota, Peltigerales, Collemataceae

Introduction

Thirty-nine species of the lichen genus Leptogium (Ach.) Gray have been

reported in China, among which 18 hairy species were recorded in Mainland

China (Wei 1991, Wang et al. 2010, Liu & Guan 2012, Liu et al. 2012, Cao et

al. 2012, Xi & Liu 2014). A recent collection of a short-haired Leptogium from

Beijing did not correspond with any named species and is described here as a

new species, L. wangii. Other herbarium specimens of short-haired Leptogium

species studied were found to represent a second new species, described here as

L. taibaiense, and a new record for China, L. burgessii.

Materials & methods

A dissecting microscope (Motic SMZ-140) and light microscope (Motic B2)

were used for the morphological and anatomical studies. Photographs were

taken with BX51 fluorescence microscope. The type specimens of Leptogium

taibaiense and L. wangii are deposited in Herbarium Mycologicum Academiae

Sinicae—Lichenes, Beijing, China (HMAS-L). Other specimens are deposited

in HMAS-L or Herbarium of Kunming Institute of Botany, Academia Sinica,

Kunming, China (KUN).

472 ... Liu & al.

Taxonomy

Leptogium taibaiense H.J. Liu & M.Q. Xi, sp. nov. PL. 1A-E

MycoBank MB 809221

Differs from Leptogium puberulum by its horizontal, entire, broader lobes and its

corticolous habitat.

Type: China. Shaanxi Province, Mt. Taibai, Temple Ping’ansi, 34°01’N 107°48’E, elev.

2700 m, on bark, 8/V1/1963, Jiang-Chun Wei 2604-1 (Holotype, HMAS-L 045039).

ErymMo oey: The species is named after the type locality, Mt. Taibai, Shaanxi, China.

THALLUS foliose, 5-6 cm in diam.; UPPER SURFACE grey green in most

parts when dry, especially near margin, other portions grey-blue to darker,

glabrous, smooth, partially minutely striate, matt; LoBEs orbicular, 5-10 mm

wide, flat, horizontal, the margins entire, downturned; Istp1A and LOBULES

absent; LOWER SURFACE paler, densely hairy, velvety, insertion of apothecia

making distinctive depressions; HAIRS 25-50 um long, composed of a row of

cylindrical cells. APOTHECIA common, + dense, laminal to submarginal, sessile

to subpedicellate, 0.2-0.5 mm in diam.; Disc plane to convex, dark red-brown;

THALLINE EXCIPLE thin, glabrous, smooth, concolorous with the thallus or

slightly paler.

THALLuS 100 um thick; CoRTEX on both sides consisting of a single layer

of subglobose, isodiametrical cells, 4-5 um in diam.; PHOTOBIONT Nostoc in

chains, spherical, 4-6 um in diam.; HYPHAE regularly interwoven. APOTHECIA

420-440 um thick; THALLINE EXCIPLE c. 80 um thick at margin, with a cortex

of a single layer of subglobose cells 4-6 um in diam., and an algal layer of

loosely interwoven hyphae; PROPER EXCIPLE subparaplectenchymatous to

somewhat euthyplectenchymatous at margin, 10-20 um thick, but indistinct

or sometimes euthyplectenchymatous in center; HYMENIUM 130-140 um high;

SUBHYMENIUM 30-40 um high; spores 8 per ascus, submuriform, ellipsoid,

18-25 x 8-12 um, 3-4-septate transversely, 1-septate longitudinally.

Corticolous.

REMARKS: Leptogium taibaiense is characterized by a combination of 1) the

glabrous and smooth upper surface and 2) the velvety lower surface with

short hairs composed of a row of cylindrical cells. Only three other species —

L. puberulum Hue, L. velutinum P.M. Jorg., L. wangii (newly described, below)

— have these characteristics. Leptogium puberulum differs from the new species

by its ascending to erect, marginally sinuous, narrower (2-5 mm) lobes, its lack

of proper exciple, its wider (14-16 um) spores, and its rock and soil substrate

(Dodge 1973, Ovstedal & Smith 2001). Leptogium velutinum differs by its bluer

upper surface, pedicellate apothecia, and larger spores (30-50 x 14-19 um;

Jorgensen 1997). Leptogium wangii (see below) differs by its pulvinate thallus,

often concave and narrower (3-5 mm wide) lobes with sinuous and upturned

Leptogium spp. nov. (China) ... 473

margins, tomentose thalline exciple, euparaplectenchymatous proper exciple,

and substrate of soil covered rocks.

Leptogium wangii H.J. Liu & J.S. Hu, sp. nov. PL. 1F-J

MycoBank MB 809222

Differs from Leptogium velutinum by its narrower lobes, sessile apothecia, and smaller

spores.

Type: China. Beijing City, Xiaolongmen National Forest Park, 39°57’N 115°26’E, elev.

1100 m, on soil covered rocks, VII/2013, Jian-Bin Chen & Xiao-Di Liu 2013-0714

(Holotype, HMAS-L 129380).

Erymo.oey: This species is named in honor of Li-Song Wang M.Sc., a lichenologist

who made numerous Leptogium collections from southwestern China.

THALLUS foliose, circular in outline, 5-6 cm in diam., pulvinate; UPPER

SURFACE grey-blue to brown when dry, glabrous, smooth to partially slightly

striate, without wrinkles, dull; Lopes 3-5 mm wide, often sub-erect, concave

to sometimes flat, the margins sinuous, upturned; Istp1a and LOBULEs absent;

LOWER SURFACE paler, velvety, densely hairy; Hairs white, (25) 40-60 (70) um

long, composed of a row of cylindrical cells. APOTHECIA rare, laminal, 0.5 mm

in diam., sessile; Disc plane to concave, dark red-brown; THALLINE EXCIPLE

smooth, white-tomentose, paler than the thallus.

THALLuS 110-130 wm thick; CoRTEX on upper side c. 10 um thick,

consisting of 1-2 layers of subglobose to + elongated, isodiametrical cells, on

lower side c. 20 um thick, consisting of 2-3 layers of subglobose to + elongated,

isodiametrical cells; PHOTOBIONT Nostoc in chains, spherical, 4-6 um in diam.;

HYPHAE Closely regularly interwoven.

APOTHECIA 300-350 um thick; THALLINE MARGIN 50-60 um thick, with a

cortex of 3-5 layers of subglobose to ellipsoid cells; HyYMENIUM 130 um high;

SUBHYMENIUM 20-30 um high; PROPER EXCIPLE euparaplectenchymatous at

margin, consisting of 3-5 layers of subglobose to ellipsoid cells, 40-60 um thick,

but often poorly developed and difficult to discern in center; LOWER CORTEX

below the apothecium 70-80 um thick, euparaplectenchymatous, consisting

of 6-8 layers of subglobose to ellipsoid cells; ascr clavate, 8-spored; sPORES

submuriform, ellipsoid, 18-25 x 8-11 um, 3 septate transversely, 1-septate

longitudinally, obtuse to slightly acute at both ends.

On soil-covered calciferous rocks.

REMARKS: Leptogium wangii is characterized by 1) a sub-erect, pulvinate

thallus; 2) a smooth, glabrous, and often concave upper surface; 3) sinuous,

ascending lobe margins; 4) short, cylindrical celled hairs on the lower surface;

5) a white tomentose apothecial margin; and 6) its habitat: soil- covered rocks.

In habitus the new species closely resembles L. puberulum, which differs by

its glabrous thalline exciples, broader spores (14-16 um wide), and absence

Leptogium spp. nov. (China) ... 475

of proper exciple (Dodge 1973, Ovstedal & Smith 2001). It also resembles

L. velutinum, which differs by its wider lobes (up to 20 mm), stalked apothecia,

and larger spores (30-50 x 14-19 um; Jorgensen 1997). For its differences from

L. taibaiense, see remarks above.

Leptogium burgessii (L.) Mont., Hist. Nat. Iles Canar. 3(2): 130 (1840). PL. 2

THALLUS foliose, 4-7 cm in diam., circular or irregular in outline; UPPER

SURFACE grey-blue, dark grey-blue to brown when dry, glabrous, smooth, dull;

LOBES horizontal, or often suberect, branched, 2-5 mm wide, rapidly narrowing

towards margin, the margins sinuous, wavy, entire to microphylline; LOwER

SURFACE paler, densely hairy; Hairs white, 10-25 um long, composed of a

chain of spherical cells. APOTHECIA few to abundant, laminal to submarginal,

0.5-5 mm in diam., sessile to subpedicellate; pisc plane to concave, red-brown

or darker; THALLINE EXCIPLE concolorous with the thallus, microphylline;

MICROPHYLLS free, abundant, occasionally fused at base in part and giving the

thalline exciple a “collar” appearance (as in HMAS-L 032520; Fic. 2H), or only

1 or 2 whorls, not forming a “collar” (as in KUN 4846; Fic. 2B).

THALLUuS 50-120 um thick; UPPER CORTEX of a single layer of subglobose,

isodiametrical cells, 5-10 um in diam.; LOWER CORTEX similar to upper cortex,

8-12 um thick; PHOTOBIONT Nostoc in chains, spherical, 3-7 um in diam.;

HYPHAE Closely regularly interwoven. APOTHECIA 400-500 um thick; THALLINE

EXCIPLE euparaplectenchymatous, at the margin consisting of 2-5 layers of

subglobose to ellipsoid cells, 50-60 um thick, medullary tissues thin or absent,

at the base with 6-12 layers of subglobose to ellipsoid, thin-walled cells,

110-150 um thick; PROPER EXCIPLE euparaplectenchymatous and continuous

with the thalline exciple laterally, consisting of 3-4 layers of subglobose to

ellipsoid cells, 40 um thick, soon becoming subparaplectenchymatous towards

center and often indistinct basally; HYMENIUM 130-200 um high; suBHYMENIUM

30-40 um high; asc clavate, 8-spored; spores submuriform, ellipsoid, 20-32 x

10-15 um, 5-6 septate transversely, 1-2-septate longitudinally, obtuse at both

ends.

Corticolous.

PLaTE 1 (left). Leptogium taibaiense (HMAS-L 045039). A. Thallus with apothecia. B. Lower

side of the thallus, showing depressions made by the insertion of apothecia, and short hairs.

C. Thallus cross-section, showing single-layered cortex on both sides, and short hairs on lower

side. D. Apothecium cross-section. E. Cross-section of apothecial margin, showing corticated

thalline margin, marginally subparaplectenchymatous proper exciple, asci and spores. Leptogium

wangii (HMAS-L 129380). E. Thallus, showing the suberect to erect lobes. G. An apothecium with

tomentose apothecial margin. H. Lower side, showing the short hairs. I. Thallus cross-section,

showing cortex, and short hairs on lower side. J. Apothecial cross-section, showing proper exciple

and the corticated thalline exciple. Scale bars: A, F = 1 cm; B = 5 mm; C = 50 um; D, E, I, J = 100

um. G=1mm;H=0.5 mm.

476 ... Liu & al.

24>

Scie

Leptogium spp. nov. (China) ... 477

SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE, Lijiang City, Yulong Snow

Mountain, 27°05’N 100°10’E, on bark, elev. 2400 m, 24/XII/1964, J.C. Wei 2744

(HMAS-L 032520). SICHUAN PROVINCE, Miyi County, Malong, Mt. Beipo, 26°58’N

101°50’E, on bark, elev. 3200 m, 7/VII/1983, L.S. Wang 83-846 (KUN 4846).

REMARKS: Leptogium burgessii is a widespread species, known from South

and North America, Europe, Macaronesia, Africa, New Zealand, and India

(Sierk 1964, Awasthi & Akhtar 1977, Swinscow & Krog 1988, Galloway 1999,

Jorgensen & Nash 2004, Aragon et al. 2005). It is new to China.

The species is characterized by smooth upper surface, short hairy lower

surface, microphylline apothecial margins, and euparaplectenchymatous

thalline exciples. Leptogium mantiqueirense Kitaura & Marcelli, L. digitatum

(A. Massal.) Zahlbr., and L. marginellum (Sw.) Gray have microphylline

appendages on the thalline margins, but can be easily separated from

L. burgessii by their distinctly wrinkled upper surfaces. Leptogium aucklandicum

Zahlbr. also has occasional microphylline outgrowths on the thalline exciples

(Galloway 1999) but differs by its glabrous lower surface.

In the smooth upper surface, microphylline to entire lobe margins, and

microphylline and euparaplectenchymatous thalline exciples, our specimens

of L. burgessii are similar to those from most literatures, especially those from

the Greater Sonoran Desert regions of the US (Jorgensen & Nash 2004). In the

euparaplectenchymatous proper exciple they resemble the type of L. inflexum

Nyl. (= L. burgessii; see Kitaura & Marcelli 2013). However, the Chinese

materials have smaller spores than those from the Greater Sonoran Desert

Region (25-50 x 14-22 um; Jorgensen & Nash 2004), from India (25-45 x

12-16 um; Awasthi & Akhtar 1977), and from Mexico (30-38 x 12-18 um, type

of L. inflexum; Kitaura & Marcelli 2013) and have larger spores than those from

New Zealand (20-25 x 7.5-10 um; Galloway 1999).

Acknowledgements

This study was supported by the National Natural Science Foundation of

China (31093440, 31000239) and Natural Science Foundation of Hebei Province

(C2014201032). The authors are indebted to Li-Song Wang M.Sc. (Kunming Institute

PLATE 2 (left). Leptogium burgessii (KUN 4846). A. Thallus with apothecia. B. Apothecia with

microphylls. C. Thallus cross-section, showing corticated surfaces, and short hairs composed of

globose cells on lower side. D. Vertical section of apothecium, showing microphylls on apothecial

margin. E. Vertical section of the apothecium showing proper exciple euparaplectenchymatous

at margin and subparaplectenchymatous towards center. F. Vertical section of thalline margin

showing the connecting thalline and proper exciples. (HMAS-L 032520). G. Thallus with apothecia.

H. Apothecia with abundant microphylls on margin. I. Thallus cross-section, showing single-

layered cortex on both sides, and short hairs composed of a row of globose cells on lower side.

J. Vertical section of apothecium, showing microphylls on apothecial margin. Scale bars: A = 1 cm;

B =2 mm; C-FE, I= 100 um; G = 2 cm; H = 5 mm; J = 200 um.

A478 ... Liu & al.

of Botany, CAS), Prof. Jiang-Chun Wei, and Ms. Hong Deng (Institute of Microbiology,

CAS) for sending the specimens on loan. The authors are grateful to Dr. Udeni Jayalal

(Department of Natural Resources, Sabaragamuwa University of Sri Lanka) and

Dr. Ze-Feng Jia (College of Life Sciences, Liaocheng University) for reading and

improving the manuscript, and for acting as presubmission reviewers.

Literature cited

Aragon G, Otalora MAG, Martinez I. 2005. New data on the genus Leptogium

(lichenized Ascomycetes) in the Iberian Peninsula. Nova Hedwigia 80(1-2): 199-226.

http://dx.doi.org/10.1127/0029-5035/2005/0080-0199

Awasthi DD, Akhtar P. 1977. The genus Leptogium (sect. Mallotium) in India. Norwegian Journal

of Botany 24: 59-71.

Cao J, Liu HJ, Deng H. 2012. A hairy species of Leptogium new to Mainland China. Journal of

Fungal Research 10(4): 213-215.

Dodge CW. 1973. Lichen flora of the Antarctic continent and adjacent islands. Phoenix Publishing

Co., Canaan, New Hampshire. 399 p.

Galloway DJ. 1999. Notes on the lichen genus Leptogium (Collemataceae, Ascomycota) in New

Zealand. Nova Hedwigia 67(3-4): 317-355.

Jorgensen PM. 1997. Further notes on hairy Leptogium species. Symbolae Botanicae Upsalienses

32: 113-30.

Jorgensen PM, Nash TH III. 2004. Leptogium. 330-350, in: TH Nash III et al. (eds). Lichen flora

of the Greater Sonoran Desert Region. Vol. 2. Lichens Unlimited, Arizona State University,

Tempe, Arizona.

Kitaura MJ, Marcelli MP. 2013. A revision of Leptogium species with spherical-celled hairs (section

Mallotium p.p.). Bryologist 116(1): 15-27. http://dx.doi.org/10.1639/0007-2745-116.1.015

Liu HJ, Guan S. 2012. A new hairy species of Leptogium (Collemataceae) from China. Mycotaxon

119: 413-417. http://dx.doi.org/10.5248/119.413

Liu HJ, Cao J, Guan S, Wu QF. 2012. Three non-hairy species of Leptogium from China. Mycotaxon

122: 483-490. http://dx.doi.org/10.5248/122.483

@Ovstedal DO, Smith RIL. 2001. Lichens of Antarctica and South Georgia: a guide to their

identification and ecology. Cambridge University Press, Cambridge. 411 p.

Sierk HA. 1964. The genus Leptogium in North America, north of Mexico. Bryologist 67(3):

245-317.

Swinscow TDV, Krog H. 1988. Macrolichens of East Africa. British Museum (Natural History).

London. 390 p.

Xi MQ, Liu HJ. 2014. Two species of Leptogium new to Asia. Journal of Fungal Research. 12(2):

71-74.

Wang HY, Ren Q, Li HM, Wang HY, Zhao ZT. 2010. Five lichens of Leptogium new to China.

Mycotaxon 111: 161-166. http://dx.doi.org/10.5248/111.161

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

278 p.

MY COTAXON

http://dx.doi.org/10.5248/130.479

Volume 130, pp. 479-482 April-June 2015

Digicatenosporium polyramosum,

a new hyphomycete from Brazil

SHEILA MIRANDA LEAO-FERREIRA', LUIS FERNANDO PASCHOLATI GUSMAO"™,

Davi AUGUSTO CARNEIRO DE ALMEIDA’, & RAFAEL FE. CASTANEDA-RUIZ?

‘Departamento de Ciéncias Biologicas, Laboratorio de Micologia, Universidade Estadual de

Feira de Santana, BR116 KM03, 44031-460, Feira de Santana, Brazil

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—Digicatenosporium polyramosum gen. & sp. nov., collected on the decaying

bark of an unidentified dicotyledonous plant in Bahia, Brazil, is described and illustrated.

This species is unique in its distinct unbranched conidiophores, monoblastic integrated

determinate conidiogenous cells, and highly digitate branched blastocatenate reddish to

yellowish brown or brown conidia.

KEY worpDs— asexual fungi, systematics, tropical fungi

Introduction

During a mycological survey of fungi associated with leaf litter in the

Brazilian Caatinga biome at Coribe, Bahia State, an interesting fungus was

collected from the decaying bark of an unidentified dicotyledonous plant. It

showed remarkable differences from all previously described hyphomycete

genera (Seifert et al. 2011) and is here proposed as a new genus and species.

Materials & methods

Samples of submerged litter were placed in paper and plastic bags, taken to the

laboratory, and prepared according to Castafieda-Ruiz (2005). Mounts were prepared

in PVL (polyvinyl alcohol, lactic acid, and phenol), and measurements were made at

a magnification of x1000. Photomicrographs were obtained with an Olympus BX51

microscope equipped with bright field and Nomarski interference optics. The type

specimen is deposited in the Herbarium of Universidade Estadual de Feira de Santana,

Bahia, Brazil (HUEFS).

480 ... Ledo-Ferreira & al.

Taxonomy

Digicatenosporium S.M. Leado, Gusmao & R.F. Castafieda, gen. nov.

MycoBank MB810773

Differs from Digitodesmium by its distinct conidiophores and branched blastocatenate

conidia.

TYPE SPECIES: Digicatenosporium polyramosum S.M. Leao et al.

EryMo_oey: From the Latin: digi- (from digitatus for finger shaped) and -catenosporium

for the conidia in chains.

Asexual fungi. COLONIES on natural substratum hairy to granulose,

ferruginous or reddish brown. Mycelium mostly immersed. CONIDIOPHORES

distinct, single, unbranched, erect, septate, pale brown. CONIDIOGENOUS CELLS

monoblastic, integrated, terminal, determinate; conidial secession schizolytic.

ConlipiA digitate or cheiroid, acrogenous, reddish brown to yellowish brown,

verruculose, central arms closely packed, marginal arms divergent, aseptate or

euseptate, subulate to slightly obclavate, some blastocatenate at the apex.

Digicatenosporium polyramosum S.M. Leao, Gusmao & RF. Castafieda, sp. nov.

MycoBank MB810774 Fic. 1

Differs from Digitodesmium spp. by having distinct conidiophores and complex

blastocatenate conidia.

Type: Brazil, Bahia, Coribe, Serra do Ramalho, 13°62’S 44°37’W, 600-800 m alt., on

decaying bark of an unidentified dicotyledonous plant, 4 March 2008, coll. $.M. Leao-

Ferreira (Holotype: HUEFS 210443).

EryMo.Loey: From the Greek poly- for many, numerous and the Latin -ramosum for

branched.

COLONIES on natural substratum hairy to granulose, ferruginous or reddish

brown. Mycelium mostly immersed. CoNIDIOPHORES distinct, single,

unbranched, erect, cylindrical, 1-2-septate, smooth, 15-40 x 3-6 um, pale

brown. CONIDIOGENOUS CELLS monoblastic, cylindrical, integrated, terminal,

determinate; conidial secession schizolytic. CoN1p1A acrogenous, digitate or

cheiroid, with 6-35 arms, overall 20-75 x 15-40 um, basal cells cuneiform,

5-6 x 3-7 um, reddish brown to yellowish brown to brown, arms paler above,

verruculose, cylindrical, subulate to slightly obclavate, 0-4-septate, yellowish

brown to reddish brown below, paler above, individually 15-30 x 5-8 um;

central arms cylindrical, closely packed, loosely incurved or parallel, dark

reddish brown or brown; marginal arms divergent, often producing an apical

secondary conidium, which may later repeatedly undergo the same process,

producing a complex chain.

Note: Digicatenosporium superficially resembles Digitodesmium P.M. Kirk

(Kirk 1981), but Digitodesmium has solitary conidia that have several to many

Digicatenosporium polyramosum gen. & sp. nov. (Brazil) ... 481

Fic. 1. Digicatenosporium polyramosum (HUEFS 210439). A. Conidiophore, conidiogenous cell,

and conidia. B. Conidia in branched chains. C-G. Conidia and secondary conidia.

482 ... Ledo-Ferreira & al.

parallel cylindrical multiseptate arms. Digicatenosporium exhibits an unusual

method for developing conidial chains in that only the apical cells of a random

number of the longer marginal arms produce a secondary conidium from the

original conidia while repetitions of this event at higher levels produce complex

branched chains.

Acknowledgments

The authors express their sincere gratitude to Prof. Bryce Kendrick and Dr. De-Wei

Li for their critical review of the manuscript. The authors are grateful to PPBio

Semiarido (CNPq/MCT]I) for financial support (Proc. 558317/2009-0) and Programa

de Pos-graduacao em Botanica (PPGBot/UEFS). The authors acknowledge the support

provided by”Programa Ciéncia sem Fronteiras”. RFCR is grateful to the Cuban Ministry

of Agriculture and “Programa de Salud Animal y Vegetal” project P131LH003033, for

facilities. We acknowledge the assistance provided by Dr. P.M. Kirk and Drs. V. Robert

and A. Stegehuis through the IndexFungorum and MycoBank websites. Dr. Lorelei

L. Norvell’s editorial and Dr. Shaun Pennycook’s nomenclatural reviews are greatly

appreciated.

Literature cited

Castanieda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Kirk PM. 1981. New or interesting microfungi II]. Dematiaceous hyphomycetes from Esher

Common, Surrey. Trans. Br. Mycol. Soc. 77: 279-297.

http://dx.doi.org/10.1016/S0007-1536(81)80031-9

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p. http://dx.doi.org/10.3767/003158511X617435

MY COTAXON

http://dx.doi.org/10.5248/130.483

Volume 130, pp. 483-488 April-June 2015

Duportella lassa sp. nov. from Northeast Asia

VIACHESLAV SPIRIN ™* & JIRE KOUT ”

‘Botanical Museum, P.O. Box 7, FI-00014 University of Helsinki, Finland

*Department of Biology, Geosciences and Environmental Education, University of West Bohemia,

Klatovska 51, Plzen, CZ-306 19, Czech Republic

* CORRESPONDENCE TO: viacheslav.spirin@helsinki.fi

Asstract — Duportella lassa is described based on 15 collections from East Siberia and

Russian Far East. The new species is characterized by its monomitic hyphal structure, brown-

colored lamprocystidia, and habit on dead branches (both attached and recently fallen) of

angiosperm trees and shrubs.

Key worps — Basidiomycota, Peniophora, Russulales, taxonomy

Introduction

Duportella Pat. (Russulales, Basidiomycota) is a small genus of the so-called

corticioid fungi currently including 13 species, mostly with tropical distribution

(Andreasen & Hallenberg 2009, Hjortstam & Ryvarden 2004). Only two species

of the genus, D. halimi (Boidin & Lang.) Hjortstam and D. malenconii (Boidin

& Lang.) Hjortstam, have been reported from temperate or subtropical areas

of the northern hemisphere (Boidin & Lanquetin 1974, 1977; Chamuris 1987;

Duhem 1990). During the field trips in Khabarovsk Reg., Russian Far East, in

2011-14, the first author collected 14 specimens of an unknown Duportella

species, and an older specimen from Siberia was found in mycological

herbarium of Botanical Museum, University of Helsinki (H). It is described

below as a new species, D. lassa.

Materials & methods

The specimens studied are kept in the herbaria of the Botanical Museum, University

of Helsinki, Finland (H), and the Mycological Herbarium of the Department of Biology,

Geosciences and Environmental Education, University of West Bohemia, Czech

Republic (KBI). The microscopic routine followed Miettinen et al. (2006) and DNA

analytical methods followed Spirin et al. (2013). Measurements were done in Cotton

Blue using phase contrast illumination and oil immersion (with a subjective accuracy

484 ... Spirin & Kout

of 0.1 um; Miettinen et al. 2006). Abbreviations used in the species descriptions (and

calculated for each specimen measured) include L - mean spore length; W - mean spore

width; Q’ - spore length/width ratio; Q - mean spore length/width ratio.

Taxonomy

Two specimens of Duportella lassa (Spirin 5498, 6129) were sequenced with

very similar results, and we found no identical or essentially close sequences

in GenBank. However, their ITS regions show up to 95% homology with some

Peniophora species, thus displaying a vague generic status for Duportella versus

Peniophora Cooke and supporting the same result shown in the complex

molecular study of Boidin et al. (1998). Although more data are needed to

resolve the phylogenetic relationships between the two taxa, for pragmatic

reasons we here accept Duportella as an independent genus distinguishable by its

brown-colored cystidia and hyphae and variably shaped basidiospores (versus

Peniophora s. str.) as well as its lack of dendrohyphidia (versus Dendrophora

(Parmasto) Chamuris); Andreasen & Hallenberg 2009).

Duportella lassa Spirin & Kout, sp. nov. PLATES 1-3

MycoBank MB 808096

D. halimii similis, sed basidiosporis minoribus.

Type: Russia, Khabarovsk Reg., Komsomol'’sk Dist., Boktor, on fallen decorticated

branch of Quercus mongolica Fisch. ex Ledeb., 18 Aug 2013, Spirin 6129 (Holotype, H;

isotype, KBI; GenBank KJ509191).

EryMo_oey: lassus (Lat., adj.): exhausted, tired; referring to the rarely found spores in

most collections of this species.

BASIDIOCARPS perennial, resupinate, covering 1-5(-10) cm, 0.05-0.15 mm

thick, first soft and waxy, then rather tough. HYMENIAL SURFACE smooth, in

juvenile basidiocarps bright ochraceous, continuous, in perennial specimens

ochraceous-brown, cracking with small irregular fissures (rimose), sometimes

with occasional low tubercles, in senescent withered basidiocarps fading to

pale brown. Margin adherent, narrow, slightly paler than hymenial surface, in

older basidiocarps indistinct. Subiculum indistinct.

HyYPHAL STRUCTURE monomitic; hyphae with clamps. Subicular hyphae

hyaline to brownish, thin- or slightly thick-walled, more or less parallel,

some tortuous and short-celled, 3-5(-6) um in diam., in older basidiocarps

producing a distinct layer 15-20 um thick. Subhymenial hyphae thin- to

slightly thick-walled, first hyaline, then brownish, irregularly and rather densely

arranged, tortuose, in subhymenium short-celled, mostly 3-4 um in diam.

LAMPROCYSTIDIA abundant, brownish to dark-brown, thick-walled, apically

strongly encrusted, ampullaceous or bottle-shaped, sometimes bifurcate, in

older specimens with 1-2 secondary septa in the middle part, 19-36 x 3-7

(-8) um. GLogocystip1A thin-walled or with thickened walls, bottle-shaped,

Duportella lassa sp. nov. (Russia) ... 485

yp rf;

SIG A!

PLATE 2. Duportella lassa (holotype), basidiocarp section. Bar = 5 um.

486 ... Spirin & Kout

PLATE 3. Duportella lassa (Spirin 5498), apically encrusted lamprocystidia (SEM). Bar = 1 um.

conical or fusiform, some bifurcate, 23-50 x 6-10.5 tm; clavate, slightly thick-

walled gloeocystidia common especially in senescent hymenium, 24-31.5 x

6.5-10.5 um. Dendrohyphidia absent. Basip1a suburniform, 4-spored, 17.5-

26 x 5-6 um, first hyaline and thin-walled, then basally thick-walled and

brownish. Basrp1ospores first hyaline, later brownish (especially in older

basidiocarps), smooth, thin-walled, thick cylindrical to narrowly ellipsoid

or ovoid, (4.6-)4.8-6.8(-7.0) x (2.7-)2.8-4.1(-4.2) um, L = 5.52, W = 3.32,

Q = (1.4-)1.5-2.1(-2.3), Q = 1.60-1.75 (n = 120/4), ventral side flat or slightly

concave, rarely obscurely convex, inamyloid, indextrinoid, acyanophilous.

ECOLOGY & DISTRIBUTION — Duportella lassa is so far the only boreal species

known in the genus. The records derive mainly from montane old growth

and secondary forests and willow thickets of East Siberia and the Russian Far

East. The holotype specimen was collected from a fallen, decorticated branch

of Quercus mongolica, but D. lassa occurs more commonly on dead but still

attached branches of Salix spp.; a few records cite other angiosperm substrates

(Acer ukurunduense Trautv. & C.A. Mey., Duschekia fruticosa (Rupr.) Pouzar,

Populus tremula L., Rhododendron dauricum L.).

ADDITIONAL SPECIMENS EXAMINED: Duportella lassa - RUSSIA. IRKUTSK REG.:

Slyudyanka Dist., Kultuk, on angiosperm, Sep 1902, Lonnbohm (Herb. P. Karsten 1514,

Duportella lassa sp. nov. (Russia) ... 487

H). KHABAROVSK RgEG.: Khabarovsk Dist., Ilga, Salix abscondita Laksch., 11 Aug 2012,

Spirin 5173 (H), Ulika, Populus tremula, 13 Aug 2012, Spirin 5220 (H), Hologu, Salix

schwerinii E.L. Wolf, 17 Aug 2012, Spirin 5340 (H, KBI), Malyi Kukachan, S. schwerinii,

20 Aug 2013, Spirin 5470 (H), Levyi Ulun, Acer ukurunduense, Salix gracilistyla Migq.,

22 Aug 2012, Spirin 5486 (H, KBI), 5498 (H, KBI; GenBank KJ509190), Rhododendron

dauricum, 24 Aug 2012, Spirin 5580 (H); Solnechnyi Dist., Chalba, Salix sp., 9 Aug

2011, Spirin 4003 (H), Suluk-Makit, Salix sp., 19 Aug 2011, Spirin 4221 (H), Sonakh,

Quercus mongolica, 15 Aug 2014, Spirin 7307, 7321 (H); Verkhnebureinskii Dist.,

Dublikan Nat. Res., Duschekia fruticosa, 19 Aug 2014 Spirin 7570 (H), S. schwerinii, 23

Aug 2014, Spirin 7949 (H).

Duportella halimi - FRANCE. LorrE-ATLANTIQUE: Le Portmain, St. Marie sur mer,

Atriplex halimus L., 31 Aug 1971, Boidin (LY-JB 6635, holotype).

Duportella kuehneri - ETHIOPIA. SHoa: Chilomo, on branch, 10 Jan 1990,

Ryvarden 28078 (H ex O).

Duportella malenconii subsp. americana Chamuris - USA. CALIFORNIA: Contra

Costa, Las Trampas Creek, on Umbellularia californica (Hook. & Arn.) Nutt., 27 Mar

1938, Bonar (H ex UC).

Duportella tristicula (Berk. & Broome) Reinking - PAKISTAN. Lahore, on dead

branches, 27 Nov 1961, Ahmad (H ex GZM).

Peniophora isabellina - USA. VirGINIA: Woodstock, Ribes sp., 20 Sep 1899, Shear

1191 (FH 00290618, holotype).

ComMENts — Duportella lassa belongs with the minority of monomitic

species in the genus. In this group, spore shape is the key character for species

identification (Andreasen & Hallenberg 2009, Boidin et al. 1991). The seemingly

most similar species, D. halima is distinguished by its broadly ellipsoid to ovoid

and larger spores [(4.8-)5.1-6.9(-7.7) x (3.7-)3.8-5.1(-5.6) um, L = 5.86,

W = 4.38, Q = (1.1-)1.2-1.4(-1.5), Q = 1.34 (n = 30/1)] and longer basidia and

cystidia (see also Duhem 1990). Moreover, D. halimi is known thus far only

from the coastal areas of France, where it inhabits dry branches and stems of

Atriplex halimus (Boidin & Lanquetin 1974, Duhem 1990).

The spores in Duportella kuehneri (Boidin & Lang.) Hjortstam are

approximately the same shape and size as in D. lassa, but it is a dimitic species

known from central Africa (Boidin & Lanquetin 1974).

The basidiocarps of Duportella lassa are usually sterile; only four specimens

(holotype, Karsten 1514, Spirin 5340, 5498) have numerous spores. Nonetheless,

it is easily recognizable even when non-fertile due to the ochraceous to brown

rimose basidiocarps, abundant dark-colored lamprocystidia, and irregularly

bifurcate gloeocystidia.

Peniophora isabellina Burt was described as possessing some characters

(basidiocarp colours, spore size, short cystidia; Burt 1925) similar to D. lassa.

Our study of the holotype (FH) revealed avery thin, pale ochraceous basidiocarp

with a smooth (not rimose) hymenial surface. Its spores, while also very scarce,

are bean-shaped (with concave ventral side) and ca. 5.3-6 x 2.8-3.1 um, and

its cystidia are carrot-shaped and encrusted with a hyaline crystalline material.

488 ... Spirin & Kout

In our opinion, these features preclude a close affinity between P. isabellina and

D. lassa.

Acknowledgments

We are very indebted to Dr. J. Vlasak (Ceské Bud&jovice, Czech Republic) for DNA

research work, J. Nebesafova and M. Mergl for the SEM photo, and curators of LY and

FH herbaria for sending us the type specimens of Duportella halimi and Peniophora

isabellina. This study was supported by the project EXLIZ - CZ.1.07/2.3.00/30.0013

(co-financed by the European Social Fund and the government of Czech Republic). The

author VS is grateful to Dr. A. Ermoshkin (Khabarovsk, Russia) for arranging the field

work in Boktor, where the holotype of D. lassa was collected. Eugene Yurchenko (Minsk,

Belarus) and Heikki Kotiranta (Helsinki, Finland) kindly revised our manuscript.

Literature cited

Andreasen M, Hallenberg N. 2009. A taxonomic survey of the Peniophoraceae. Syn. Fungorum

26: 56-119.

Boidin J, Lanquetin P. 1974. Peniophora (sect. Duportella) kuehneri et halimi novae sp.; réflexions

sur les genres Peniophora et Duportella. Bull. Mens. Soc. Linn. Lyon (special issue): 47-60.

Boidin J, Lanquetin P. 1977. Peniophora (sect. Duportella) malengonii [sic] nov. sp. (Basidiomycetes

Corticiaceae), espece méditerranéenne partiellement interstérile avec son vicariant californien.

Revue Mycol., Paris 41: 119-128.

Boidin J, Lanquetin P, Gilles G. 1991. Les Peniophoraceae de la zone intertropicale (Basidiomycetes,

Aphyllophorales). Bull. Soc. Mycol. France 107(3): 91-156.

Boidin J, Mugnier J, Canales R. 1998. Taxonomie moleculaire des Aphyllophorales. Mycotaxon 66:

445-491.

Burt EA. 1925. The Thelephoraceae of North America. 14. Peniophora. Ann. Missouri Bot. Gard.

12213-3957:

Chamuris G. 1987. Notes on stereoid fungi 1. The genus Dendrophora, stat. nova, and Peniophora

malenconii subsp. americana, subsp. nova (“Stereum heterosporum”). Mycotaxon 28: 543-552.

Duhem B. 1990. Etudes de cing Peniophora récoltés sur Pile de Noirmoutier. Doc. Mycol. 20(78):

11-34.

Hjortstam K, Ryvarden L. 2004. Some new genera and species of corticioid fungi (Basidiomycotina,

Aphyllophorales) from tropical areas. Syn. Fungorum 18: 20-32.

Miettinen O, Niemela T, Spirin V. 2006. Northern Antrodiella species: the identity of Antrodiella

semisupina, and type studies of related taxa. Mycotaxon 96: 211-239.

Spirin V, Vlasak J, Niemela T, Miettinen O. 2013. What is Antrodia sensu stricto? Mycologia 105(6):

1555-1576. http://dx.doi.org/10.3852/13-039

MYCOTAXON

http://dx.doi.org/10.5248/130.489

Volume 130, pp. 489-493 April-June 2015

Brachycephala exotica,

a new hyphomycete from Brazil

JOSIANE SANTANA MONTEIRO’, Luis FERNANDO PASCHOLATI GUSMAO’,

& RAFAEL E. CASTANEDA-RUIZ?

‘Universidade Estadual de Feira de Santana, Departamento de Ciéncias Bioldgicas,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Bahia, Brazil

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P.17200

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT —Brachycephala exotica gen. & sp. nov., is described and illustrated. Brachycephala

exotica is characterized by distinct single conidiophores and monoblastic conidiogenous cells

that produce solitary acrogenous 2-3-septate dark subglobose conidia with 1-3 branches that

arise from the basal conidial cell to form scorpioid branched conidiophores.

KEY worps — asexual fungi, systematics, leaf litter, tropical fungi

Introduction

During investigations of microfungi on dead plant material in a semi-arid

region of northeast Brazil an interesting fungus was collected on decaying

leaves of an unidentified plant. It showed remarkable differences from all

previously described hyphomycete genera (Seifert et al. 2011) and is therefore

described here as new.

Materials & methods

Samples of submerged litter were placed in paper bags, taken to the laboratory, and

treated according to Castafteda-Ruiz (2005). Mounts were prepared in PVL (polyvinyl

alcohol, lactic acid, and phenol) and measurements were taken at x1000. Micrographs

were obtained with an Olympus microscope (model BX51) equipped with bright field

and Nomarski interference optics. The holotype is deposited in the Herbarium of

Universidade Estadual de Feira de Santana, Bahia, Brazil (HUEFS).

490 ... Monteiro, Gusmao, & Castafieda-Ruiz

Taxonomy

Brachycephala J.S. Monteiro, Gusmao & R.F. Castafieda, gen. nov.

IF550848

Differs from Monotosporella by its scorpioid, branched conidiophores and rhexolytic

conidial secession and from Brachysporiella and Melanocephala by its scorpioid,

branched conidiophores.

TyPeE SPECIES: Brachycephala exotica J.S. Monteiro et al.

EryMoLoGcy: From the Greek: brachy-, referring to the branched conidiophores +

-cephala, referring to the head-shaped conidia.

Asexual fungi. CoLtonigs on the natural substratum, hairy to funiculose, black.

Mycelium superficial and immersed. ConiDIOPHORES of two types: i) distinct,

single, flexuous, unbranched below, branching at the basal cell of the conidium,

cylindrical, septate, smooth, brown to dark brown; ii) secondary, tertiary,

quaternary, distinct, straight or slightly flexuous to somewhat curved, septate,

smooth, pale brown, arising from the basal cell of each conidium, forming

scorpioid branches. CONIDIOGENOUS CELLS monoblastic, integrated, terminal,

determinate or indeterminate with several percurrent extensions, cylindrical,

pale brown. Conidial secession rhexolytic. Conrp1a solitary, acrogenous,

septate, pyriform to obovoid to globose, brown, dark brown or black, smooth,

often branching at the apex of the basal cell.

Brachycephala exotica J.S. Monteiro, Gusmao & R.E. Castafieda, sp.nov. Figs 1, 2

IF550849

Differs from Brachysporiella, Monotosporella, and Melanocephala spp. by scorpioid

branched conidiophores originating at the basal cell of the conidium.

TyPE: Brazil, Ceara State, Quiixada, Reserva Particular do Patriménio Natural Frazenda

Nao Me Deixes, 4°49 S 38°58 W, on decaying leaves of an unidentified plant, 1 April

2014, coll. D.S. Alfredo & J.O. Souza (Holotype: HUEFS 210446).

ErymMo_oey: From the Latin: exotica, referring to the unusual or uncommon branched

conidiophores.

Cotonigs on the natural substratum, hairy to funiculose, black. Mycelium

superficial and immersed. CONIDIOPHORES of two types: i) distinct, single,

flexuous, unbranched below, but branched at the basal cell of the conidium near

its apex, 75-200 x 4-7 um, cylindrical, inflated at the base, 4-7 septate, smooth,

brown to dark brown below, pale brown toward the apex; ii) secondary, tertiary

or quaternary conidiophores, distinct, straight or slightly flexuous to somewhat

curved, 0-2-septate, smooth, pale brown, 15-50 x 2-4 um, arising from the

basal cell of each conidium, forming scorpioid branches. CONIDIOGENOUS

Fic. 1. Brachycephala exotica (holotype, HUEFS 210446). A. Conidium and conidiogenous cell.

B-L. Conidia. M-O. Conidiophores, conidiogenous cells, and conidia. Scale bars: A-L = 10 um;

M-O = 20 um.

Brachycephala exotica gen. & sp. nov. (Brazil) ... 491

492 ... Monteiro, Gusmao, & Castafieda-Ruiz

Fic. 2. Brachycephala exotica (holotype, HUEFS 210446). A-D. Conidia and branches formed

in the basal cell. E-F. Conidia and secondary conidiophores. G-K. Conidia and secondary and

tertiary conidiophores in scorpioid branches. Scale bars = 10 um.

Brachycephala exotica gen. & sp. nov. (Brazil) ... 493

CELLS monoblastic, integrated, terminal, determinate or indeterminate,

with several percurrent extensions, cylindrical to subulate, 10-20 x 3-6 um,

pale brown. Conidial secession rhexolytic. Conrp1A solitary, acrogenous

1-3-septate, mostly 2-septate. pyriform to obovoid to globose, 20-37 x 8-16

um, dark brown or black at the apical cell, pale brown to brown lower cells,

smooth, often with 1-3 branches arising from the upper part of the basal cell.

Note: The scorpioid branches arising from the basal cell of the conidia of

Brachycephala are unique among hyphomycetes (Seifert et al. 2011) and are

similar only to those of some members of the Mucoraceae (Zygomycota).

Brachycephala superficially resembles Acarocybellina Subram., Brachysporiella

Bat., and Kramabeeja G.V. Rao & K.A. Reddy, but in these genera the branches

arise from the axis of the conidiophores (Seifert et al. 2011).

Acknowledgments

The authors express their sincere gratitude to Prof. Bryce Kendrick and Dr. De-Wei

Li for their critical review of the manuscript. The authors are grateful to the “Programa

de Pesquisa em Biodiversidade” - (PPBio Semi-arid/MCTI/CNPgq) for financial

support. The authors thank the support provided by “Programa Ciéncia sem Fronteiras.”

RFCR is grateful to Cuban Ministry of Agriculture and “Programa de Salud Animal y

Vegetal”, project P131LH003033 for facilities. We acknowledge the assistance provided

by Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the IndexFungorum

and MycoBank websites. Dr. Lorelei Norvell’s editorial and Dr. Shaun Pennycook’s

nomenclatural reviews are greatly appreciated.

Literature cited

Castafieda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia. Brasilia.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes.

CBS Biodiversity Series 9. 997 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.495

Volume 130, pp. 495-498 April-June 2015

A new species of Diplococcium

from the Brazilian semi-arid region

Davi AUGUSTO CARNEIRO DE ALMEIDA’, TASCIANO DOS SANTOS SANTA IZABEL’,

Luis FERNANDO PASCHOLATI GUSMAO?*, & RAFAEL E CASTANEDA-RUIZ?

Universidade Estadual de Feira de Santana, Departamento de Ciéncias Bioldgicas,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Bahia, Brazil

?Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

* CORRESPONDENCE TO: lgusmao@uefs. br

AxsstRacT — An undescribed hyphomycete collected during investigations of microfungi

on dead plant material in a semi-arid region of northeast Brazil is described and illustrated

as Diplococcium heteroseptatum sp. nov. The new fungus is distinguished by differentiated

brown verruculose conidiophores and oblong to cylindrical 0-12-heteroseptate (but mostly

1-septate) brown conidia.

Key worps— biodiversity, taxonomy, tropical microfungi

Introduction

During a mycological survey of conidial fungi from the semi-arid region in

Bahia State, Brazil, a conspicuous fungus was collected that is clearly related to

the genus Diplococcium Grove, introduced by Grove (1885) with D. spicatum

as the type species. The genus comprises 29 species characterized by distinct,

erect or funiculose, branched, pigmented conidiophores and blastocatenate,

unicellular or septate, pigmented conidia produced on polytretic conidiogenous

cells (Silva et al. 2014).

Materials & methods

Samples of bark were collected from March 2007 to November 2008 at Serra

da Fumaca (10°74’S 40°36’W) and Serra da Jibdéia (12°50’S 39°28’W), Bahia state.

Methods for specimen collection, isolation, and morphological examination are

provided in Almeida et al. (2014). Holotype and additional specimens were deposited

in the Herbarium of the State University of Feira de Santana, Bahia, Brazil (HUEFS).

496 ... Almeida & al.

Toa

K L

TOO anos Dob

a 20pm 2m 7m

Fic. 1. Diplococcium heteroseptatum (holotype, HUEFS 210432): A-B. General aspect.

C-D. Conidiophores, conidiogenous cells, and conidia. E-L. Conidia showing the variation in the

number of septa.

Diplococcium heteroseptatum sp. nov. (Brazil) ... 497

Taxonomy

Diplococcium heteroseptatum D.A.C. Almeida, T.S. Santa Izabel,

R.F. Castafieda & Gusmao, sp. nov. Fics 1, 2

MycoBank MB810699

Differs from all other Diplococcium spp. by the wide range of conidial septation.

Type: Brazil, Bahia State, Pindobacu, Serra da Fumaga, on bark of an unidentified plant,

20.X1.2008, coll. D.A.C. Almeida (Holotype: HUEFS 210432).

ErymMo_oey: from the Greek, hetero-, referring to the variable numbers of septa + from

the Latin, -septatum, referring to the septa.

CONIDIOPHORES distinct, single or grouped to profusely fasciculate, erect,

unbranched or rarely branched, straight or flexuous, septate, verrucose or

verruculose below, smooth toward the apex, brown, 120-300 x 4.5-9 um.

CONIDIOGENOUS CELL polytretic, integrated, terminal, smooth, brown.

ConipiA blastocatenate, ellipsoid to oblong or cylindrical, rounded at the ends,

0-12-septate, but mostly 1-septate, constricted at the somewhat thickened

septa, smooth, brown, concolorous, 10-77 x 6-10 um.

ADDITIONAL SPECIMENS EXAMINED: BRAZIL, BAHIA, Serra da Jibdia, on bark of an

unidentified plant, 27.11.2007, coll. T.S. Santa-Izabel (HUEFS 210428; 210429; 210430;

210430); 9.IV.2007, coll. T.S. Santa-Izabel (HUEFS 210431).

Note: Diplococcium heteroseptatum also differs from all other Diplococcium

species by its profusely fasciculate, verrucose or verruculose conidiophores and

the random number of conidial septa observed.

Acknowledgments

The authors thank Dr. Prof. Bryce Kendrick and Dr. De-Wei Li for critically

reviewing the manuscript. The authors thank the PPBIO Semi-arid (MCTI -CNPq/

proc. 554718/2009-0) for financial support. The first and second authors thank the

(CAPES) and (CNPq) for scholarships. LFPG thanks CNPq (proc. 305413/2011-2).

RFCR is also grateful to the Cuban Ministry of Agriculture and “Programa de

Salud Animal y Vegetal’, project P131LH003033, for facilities. We acknowledge the

assistance provided by Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the

IndexFungorum and MycoBank websites. Dr. Lorelei Norvell’s editorial and Dr. Shaun

Pennycook’s nomenclatural reviews are greatly appreciated.

Literature cited

Almeida DAC, Miller AN, Gusmao LFP. 2014. New species and new combinations of

conidial fungi from semi-arid Caatinga biome of Brazil. Nova Hedwigia 98: 431-447.

http://dx.doi.org/10.1127/0029-5035/2013/0162

Grove WB. 1885. New or noteworthy fungi. II. Journal of Botany British and Foreign. 23: 155-168.

Silva SS, Cruz ACR, Gusmao LFP, Castafieda-Ruiz RFE. 2014. Diplococcium variegatum a

new conidial fungi from semi-arid Caatinga biome of Brazil. Mycotaxon 127: 59-62.

http://dx.doi.org/10.5248/127.59

498 ... Almeida & al.

Fic. 2. Diplococcium heteroseptatum (holotype, HUEFS 210432):

A: Conidiophores, conidiogenous cells, and conidia. B: Conidiogenous cell and conidia.

MY COTAXON

http://dx.doi.org/10.5248/130.499

Volume 130, pp. 499-503 April-June 2015

Distophragmia, a new genus of microfungi to accommodate

Endophragmiella rigidiuscula

RAFAEL FE. CASTANEDA-RUIZ', SHEILA MIRANDA LEAO-FERREIRA?,

& Luis FERNANDO PASCHOLATI GUSMAO”*

"Instituto de Investigaciones Fundamentales en Agricultura Tropical ‘Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

?Universidade Estadual de Feira de Santana, Departamento de Ciéncias Biol6gicas,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Bahia, Brazil

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—Distophragmia gen. nov. is established to accommodate Endophragmiella

rigidiuscula based on the type of conidial septa. Descriptions and illustrations are provided.

KEY worDs— asexual fungi, systematics, tropical fungi

Introduction

Endophragmiella B. Sutton (Sutton 1973) is characterized by monoblastic,

integrated, terminal, indeterminate, conidiogenous cells with several or

numerous enteroblastic percurrent extensions and unicellular or euseptate

pigmented conidia that secede rhexolytically. Endophragmiella rigidiuscula

shares these common characters of Endophragmiella species, except that its

conidial septa are distosepta (Castafieda-Ruiz 1988, Ledo-Ferreira et al. 2009).

If species with distoseptate conidia were included in Endophragmiella, the

generic concept would have to be expanded drastically. The distoseptate conidia

provide good reason to regard Endophragmiella rigidiuscula as representing a

separate genus. For that reason we propose Distophragmia as a new genus.

Taxonomy

Distophragmia R.F. Castafieda, S.M. Leaéo & Gusmio, gen. nov.

MycoBank MB810799

Differs from Endophragmiella by its distoseptate conidia.

500 ... Castafteda-Ruiz, Leao-Ferreira, & Gusmao

Type species: Endophragmiella rigidiuscula R.F. Castafeda [= Distophragmia

rigidiuscula]

ErymMo_oey: From the Latin disto- (from distitutus), referring to the type of septa + the

Greek -phragmia, referring to the phragmospore.

Asexual fungi. CoLonigs on the natural substratum hairy, brown to black.

Mycelium mostly immersed. ConipIopHoRES distinct, single, unbranched,

erect, septate, brown. CONIDIOGENOUS CELLS monoblastic, integrated, terminal,

determinate or indeterminate, with several enteroblastic percurrent extensions.

Conidial secession rhexolytic. Conrp1a solitary, acrogenous, ellipsoid, obovoid

to globose, distoseptate, smooth, brown.

Note: Sporidesmiella P.M. Kirk resembles Distophragmia, but although

Sporidesmiella shares conidia with a similar conidial ontogeny that are

disto- and euseptate or only distoseptate, its schizolytic conidial secession

is diagnostic (Kirk 1982). Matsushimiella R.E. Castafeda & Heredia, which

superficially resembles Distophragmia, is separated by sympodial extensions

of its conidiogenous cells and separating cells (Castafteda-Ruiz et al. 2001).

In Distophragmia, sometimes a cupulate remnant of the detached outer layers

remains on the conidiogenous cell walls after each enteroblastic percurrent

extension, a character shared by Endophragmiopsis M.B. Ellis, Melanocephala

S. Hughes, and Phragmocephala E.W. Mason & S. Hughes, genera that are

distinguished from Distophragmia by their euseptate conidia (Seifert et al

2011).

Distophragmia rigidiuscula (R.F. Castafieda) R.F. Castafieda, S.M. Ledo

& Gusmao, comb. nov. Fics 1,2

MycoBank MB810800

= Endophragmiella rigidiuscula R.F. Castafeda, Fungi Cubenses 3: 9 (1988)

COLONIES on the natural substratum, hairy, black. Mycelium mostly immersed,

composed of branched, septate, 2-4 um wide, smooth, brown to dark brown

hyphae. CoNnrIDIOPHORES distinct, single, unbranched, erect, straight,

cylindrical, rigid, 40-100 x 4-6 um, 1-2-septate, smooth, 2-4 percurrent

elongations, brown below, pale brown toward the apex. CONIDIOGENOUS CELLS

monoblastic, cylindrical, integrated, terminal, determinate or indeterminate

with percurrent extensions. Conidial secession rhexolytic. Conrp1A solitary,

acrogenous, ellipsoid, obovoid to globose, 2-3-distoseptate, brown, lumina

dark brown at the distosepta, 20-23 x 13-15 um, smooth, with a cylindrical

basal frill.

SPECIMENS EXAMINED: CUBA, PINAR DEL RIO PROVINCE, Vifiales Valley, 22°36’N

83°42’W, on decaying leaves Byrsonima crassifolia (L.) Kunth, 25 November 1987, coll.

R.E Castaneda-Ruiz (INIFAT C87/320, holotype). BRAZIL. Banta, Santa Terezinha,

Serra da Jibdia, 15°50’S 39°28’W, on decaying leaves of Andira fraxinifolia Benth., 5

April 2005; coll. S.M. Leao-Ferreira (HUEFS 98004).

Distophragmia gen. nov. for Endophragmiella rigidiuscula ... 501

5 um

Fic. 1. Distophragmia rigidiuscula (HUEFS 98004).

A. Conidiophores, conidiogenous cells, and conidia. B. Conidia.

502 ... Castafteda-Ruiz, Leao-Ferreira, & Gusmao

5 um

Fic. 2. Distophragmia rigidiuscula (HUEFS 98004).

A. Conidia. B. Conidiophores, conidiogenous cells, and conidia.

Distophragmia gen. nov. for Endophragmiella rigidiuscula ... 503

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li and Prof. Dr. Ze-Fen

Yu for their critical review of the manuscript. The authors are grateful to the PPBIO

Semi-arid (MCTI - CNPq/proc. 554718/2009-0) for financial support. The authors

acknowledge the support provided by “Programa Ciéncia sem Fronteiras”. RFCR is

grateful to the Cuban Ministry of Agriculture and “Programa de Salud Animal y Vegetal”

project P131LH003033, for facilities. We also acknowledge the assistance provided by

Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the IndexFungorum and

MycoBank websites. Dr. Lorelei Norvell's editorial and Dr. Shaun Pennycook’s

nomenclatural reviews are greatly appreciated.

Literature cited

Castafieda-Ruiz RE. 1988. Fungi cubenses III. Inst. Invest. Fund. Agric. Trop. ‘Alejandro de

Humboldt: 27 p.

Castafieda-Ruiz RF, Heredia G, Reyes M, Arias RM, Decock C. 2001. A revision of the genus

Pseudospiropes and some new taxa. Cryptogamie Mycol. 22: 3-18.

http://dx.doi.org/10.1016/S0181-1584(01)01057-0

Kirk PM. 1982. New or interesting microfungi VI. Sporidesmiella gen. nov. (hyphomycetes). Trans.

Br. Mycol. Soc. 79:479-489. http://dx.doi.org/10.1016/S0007-1536(82)80040-5

Leao-Ferreira SM, Santa-Izabel TS, Gusmao LFP, Marques MFO. 2009. Novas ocorréncias

de fungos conidiais para América do Sul e neotrdpico. Revta Brasil. Bot. 32: 775-780.

http://dx.doi.org/10.1590/S0100-84042009000400015

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycol. Pap. 132.

143 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.505

Volume 130, pp. 505-509 April-June 2015

A new species of Chaetochalara on decaying leaves from Brazil

SILVANA SANTOS DA SILVA‘, CAROLINA RIBEIRO SILVA ”,

Luis FERNANDO PASCHOLATI GUSMAO’ , & RAFAEL FE. CASTANEDA- RUIZ}

' Universidade Estadual de Feira de Santana, Departamento de Ciéncias Biologicas,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Bahia, Brazil

? Departamento de Micologia, Universidade Federal de Pernambuco,

50670-420, Recife, PE, Brazil

*Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt’ (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P.17200

*CORRESPONDENCE TO: lgusmao@uefs. br

AxsstrRact —Chaetochalara mutabilis sp. nov., found on decaying needle-like leaves of

Araucaria angustifolia and leaves of Calophyllum brasiliense, is described and illustrated. The

new species is characterized by erect septate dark brown setae (most of which develop into

distinct single dark brown conidiophores) and cylindrical 1-septate hyaline conidia. A key to

Chaetochalara species is also provided.

KEY worps — asexual fungi, leaf litter, taxonomy, tropical fungi

Introduction

During research on conidial fungi associated with decaying leaves of

Araucaria angustifolia and Calophyllum brasiliense, an interesting fungus

was collected. Its setae, conidiogenesis, and conidial features clearly suggest

placement within the genus Chaetochalara B. Sutton & Piroz. (Nag Raj &

Kendrick 1975, Taylor et al. 2001). However, the fungus shows remarkable

differences from all previously described species of Chaetochalara and therefore

is described as new.

Materials & methods

Samples of decaying leaves of A. angustifolia and C. brasiliense were collected and

placed in paper bags. In the laboratory the samples were placed in Petri dish moist

chambers and stored in a polystyrene box with sterile water plus glycerol at 25°C for 30

days (Castafieda-Ruiz 2005). Mounts of slides were prepared in PVL (polyvinyl alcohol,

506 ... Silva & al.

lactic acid, and phenol), and micrographs were obtained with an Olympus microscope

BX 51. Specimens were deposited in the Herbarium of the State University of Feira de

Santana, Bahia, Brazil (HUEFS).

Taxonomy

Chaetochalara mutabilis C.R. Silva, S.S. Silva, Gusmao &

R.F. Castaneda, sp. nov. Fig. 1

MycoBank MB810746

Differs from all other Chaetochalara spp. by its dimorphic conidiogenous cells and its

changeable setae, most of which develop into distinct unbranched multiseptate dark

brown conidiophores.

Type: Brazil. Bahia State: Piaté, Serra da Tromba, 13°05’S 41°50’W, alt. 1263 m, on

decaying leaves of Calophyllum brasiliense Cambess. (Calophyllaceae), 17.11.2014, coll.

C.R. Silva (Holotype: HUEFS 210434).

ErymMoLoecy: Latin, mutabilis, referring to the changeable setae, which become

conidiophores.

CoLoniEs on the natural substrate effuse, hairy, brown. Mycelium superficial

and immersed. SETAE erect, straight or slightly flexuous, cylindrical or

subcylindrical, 5—14-septate, 150-345 x 4-8 um, dark brown, pale brown

toward the apex, most developing into a distinct, unbranched, dark brown

conidiophore. CONIDIOPHORES distinct, single, erect, straight to slightly

flexuous, unbranched, associated with the base of setae, ampulliform, pale

brown, 1-3-septate, 40-82 x 4-8 um, smooth. CONIDIOGENOUS CELLS

phialidic, dimorphic: i) obclavate to subcylindrical, pale brown, 42-58 um

long; venter subcylindrical, 17—22 x 4—8 um; collarette cylindrical 25-37 x 3

um; gradual transition from venter to collarette; ii) ampulliform, pale brown,

24—57um long, venter subcylindrical, 12-25 x 4-6 um, collarette cylindrical

12-33 x 3 um, gradual transition from venter to collarette, arising from

transformed conidiophores and originating at the apical cell of the setae.

Conipia basocatenate, cylindrical, slightly rounded at the apex and truncate at

the base, 1-septate, hyaline, smooth, 10-15 x 2—2.5 um.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO SUL STATE: Sao

Francisco de Paula, Floresta Nacional de Sao Francisco de Paula 29°25’S 50°23’W,

alt. 838 m, on decaying needle-like leaves of Araucaria angustifolia (Bertol.) Kuntze

(Araucariaceae), 9.1X.2014; coll. S.S. Silva (HUEFS 210433).

Notes: Chaetochalara was erected by Sutton & Pirozynski (1965) with the

type species C. bulbosa B. Sutton & Piroz. The genus, which is distinguished

Fic. 1. Chaetochalara mutabilis (holotype, HUEFS 210434). A-B. Apex of setae. C—-E. Transformed

setae, originating conidiogenous cells at the apex of conidiophores (type ii). F-G. Conidia.

H-I. Setae. J. General aspect of setae, conidiophores and conidiogenous cells (types i & ii).

K-L. Conidiophores. M-N. Conidiogenous cells (type i). Scale bars: A-E=10 um; F-N=20 um.

Chaetochalara mutabilis sp. nov. (Brazil) ... 507

B C D E F

Eas

10 pm

508 ... Silva & al.

from Chalara (Corda) Rabenh. by the presence of setae, currently comprises

eight species (Sutton & Pirozynski 1965, Pirozynski & Hodges 1973, Sutton &

Hodges 1976, Taylor et al. 2001). Chaetochalara ramosa Nag Raj & W.B. Kendr.

and C. setosa (Harkn.) Nag Raj & W.B. Kendr. resemble C. mutabilis in their

conidial septation, but C. ramosa has a marginal conidial frill, C. setosa has

an abrupt transition from venter to collarette, and both species have shorter

(25-38 um) phialides.

Key to Chaetochalara species

I Setae not-developitioainto-conidtophiGres sac. haa Foo oa cd has oh hoa ce hon Sewiaa han A 2

1. Setae mostly developing into distinct cylindrical 5-14-septate conidiophores;

phialides dimorphic, smooth, either ampulliform (42—58 um long)

or obclavate to subcylindrical (24-57 um long); conidia cylindrical,

Isseptate; LOA IS Se ZA DSU... Fain eg dine eda ater d4 herd herrda Hartas-sit C. mutabilis

2, ell conidia im icel MAL of .%. ba dra oesea dees seton dbs gebog dis peked duis debe edit geben dyin prbmog be folourarin felonds 3

Def ll OF SOUPS CONIC IA SED EALE ¢ ne eas orate tare ee oe ote te ane eee Beg eo Bee a -:

3. Phialides narrowly ampulliform, smooth, 21-37 um long;

Gti Rae S HOA SSO, reasteeesera cit esecn taitesecs saat etgarahet ge uate Saat ot C. africana

3. Phialides ampulliform, smooth, 21—37 um long;

TREINNich CMe Moseat LOhy co Ia seneay CRT ey Ai Ran: CSP Ee Semen Or ee oie dea eee C. bulbosa

4. Conidia 0-1-septate, 10-18 x 4—5 um; phialides subcylindrical to obclavate,

SOS 45 Mi LOT OST OO ENT e Bay Bessette lalate sole sd pee Ace Gp igi tcn ees de C. cladii

A LCOMICTASCPLALEN.... aides gine, pour, BAU Ney IG RLY WEN Ny BEOUN RA OL ee epee MO ee a 5

SaPhialideseyertuculose:.t.c! wee tee. eee ne Oe, eR EN Ee nn ee eee a: 6

Dre MVANTACS SHO OTN 9.4: ociea-g upton dbo scders dri ontegstty Peay db Pion chis soaeg cdr sebgecdr hp tanh peteuneyes peeled 7

6. Phialides ampulliform, 48—77 um long; conidia 10-27 x 2.5-4um........ C. aspera

6. Phialides subcylindrical, 70-90 um long; conidia 9-14 x 5-7 um........ C. proteae

7. Phialides arising as branches of short conidiophores, subcylindrical,

25-38 tiimlones Conidia:9 14 3g 3— 4. Sine: iva ea tanta savas beawnrhs, Seavarche C. ramosa

7. Phialides not arising as branches of short conidiophores...................0.06. 8

8. Phialides ampulliform, 25-38 um long; conidia 11-15 x 1.5-2.5um...... C. setosa

8. Phialides long lageniform, 80-100 um long; conidia 16.5—-19 x 2.5-3 um .. C. laevis

Acknowledgments

The authors express their sincere gratitude to Dr. De- Wei Liand Dr. Shambhu Kumar for

their critical review of the manuscript. The authors thank the Cnpq (Proc:141475/2013-7),

“Programa de Pds-Graduacao em Botanica (PPGBot/ UEFS)’,“Programa de Pés-

Graduacgao em Biologia de Fungos (PPGBF/ UFPE)’, “Programa de pesquisa em

Biodiversidade do Semiarido” (PPBIO semi-arid - MCTI/CNPgq proc. 554718/2009-0).

RFCR is grateful to The Cuban Ministry of Agriculture and “Programa de Salud Animal

y Vegetal’, project P131LH003033 for facilities. We acknowledge the facilities provided

Chaetochalara mutabilis sp. nov. (Brazil) ... 509

by Dr. P.M. Kirk and Drs. V. Robert and A. Decock through the IndexFungorum and

MycoBank websites. Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s

nomenclature review are greatly appreciated

Literature cited

Castaneda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in: Anais

do V Congresso Latino Americano de Micologia, Brasilia.

Nag Raj TR, Kendrick B. 1975. A monograph of Chalara and allied genera. Wilfrid Laurier

University Press Waterloo, Ontario, Canada.

Pirozynski KA, Hodges CS. 1973. New hyphomycetes from South Carolina. Can. J. Bot. 51:

157-173. http://dx.doi.org/10.1139/b73-024

Sutton BC, Hodges CS. 1976. Eucalyptus microfungi: some setose hyphomycetes with phialides.

Nova Hedwigia 27(12): 343-352.

Sutton BC, Pirozynski KA. 1965. Notes on microfungi. II. Trans. Br. Mycol. Soc. 48(3): 349-366.

http://dx.doi.org/10.1016/S0007-1536(65)80055-9

Taylor JE, Crous, PW, Palm ME. 2001. Foliar and stem fungal pathogens of Proteaceae in Hawaii.

Mycotaxon. 78: 449-490.

MY COTAXON

http://dx.doi.org/10.5248/130.511

Volume 130, pp. 511-516 April-June 2015

Two new species of Spadicoides and Sporidesmiella

from Yucatan, Mexico

GABRIELA HEREDIA’, MARCELA GAMBOA-ANGULO’,

Rosa M. ARIAS’, & RAFAEL F. CASTANEDA-RUIZ?

‘Instituto de Ecologia A.C., Carretera antigua a Coatepec No. 351,

Congregacion El Haya, 91070 Xalapa, Veracruz, México

*Centro de Investigacion Cientifica de Yucatan A.C.,

Calle 43 No.130, Chuburna, Mérida, 97200, Yucatan, México

?Instituto de Investigaciones Fundamentales en Agricultura Tropical ‘Alejandro de Humboldt' (INIFAT),

Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

*CORRESPONDENCE TO: gabriela. heredia@inecol.mx

ABSTRACT — Spadicoides sylvatica sp. nov. and Sporidesmiella mammillata sp. nov., both

collected on decaying plant material in a Mexican dry tropical forest, are described and

illustrated. Spadicoides sylvatica is distinguished by its ovate to obclavate, 3—4-septate,

smooth, golden brown to brown conidia. Sporidesmiella mammillata is characterized by

conidia that are solitary acrogenous, obclavate to fusiform, sub-rostrate, 4—7-distoseptate,

smooth, and golden brown.

KEY worpDs — asexual fungi, systematics, tropical fungi

Introduction

Saprobic dematiaceous hyphomycetes are highly diverse on plant material

in tropical forests, where many new genera or species have recently been

discovered (e.g., Castaneda Ruiz et al. 2009, 2012; Heredia et al. 2013, 2014;

Zhang et al. 2009, 2011). During a mycological survey of fungi associated with

decaying plant material from a Mexican dry tropical forest, two interesting

fungi were collected that showed remarkable differences from all previously

described Spadicoides and Sporidesmiella species. The two fungi are described

here as new.

Materials & methods

Samples of plant debris (twigs, bark, leaves, and petioles) were collected in paper

bags. In the laboratory the samples were placed in Petri dish moist chambers and stored

512 ... Heredia & al.

in a 170 L polystyrene box with 200 mL sterile water plus 2 mL glycerol at 25°C for 30

days (Castafieda-Ruiz 2005). During this period, the samples were examined for the

presence of conidial fungi. Slides were prepared in polyvinyl alcohol and lactic acid.

Measurements were made at a magnification of x1000. Micrographs were obtained with

a Nikon eclipse 80i microscope equipped with bright field and Nomarski interference

optics. The type specimens are deposited in the Herbarium of the Instituto de Ecologia

A. C., Xalapa, Mexico (XAL).

Taxonomy

Spadicoides sylvatica Heredia, R.F. Castafieda & R.M. Arias, sp. nov. Fic. 1A-c

MycoBank MB812922

Differs from Spadicoides yunnanensis by its ovate to obclavate, 3—4-septate, longer, and

narrower conidia.

Type: Mexico, Yucatan State, Oxkutzcab municipality, Kaxil Kiuic Biocultural Reserve,

20°17’N 89°23’W, dry tropical forest, on decaying twig, 28 January 2014, coll. G. Heredia

(Holotype: XAL CB1687).

Erymo oey: sylvatica, refers to woodland or forest.

CoLonigs on the natural substrate effuse, hairy, dark brown to brown.

Mycelium mostly immersed, composed of septate, branched, brown, smooth

hyphae. Conip1opHores differentiated, mononematous, simple, erect, straight,

5-8-septate, smooth, dark brown, 180-220 x 6-9 um. CONIDIOGENOUS CELLS

polytretic, integrated, terminal and intercalary, cylindrical, brown, 13-28 x

5-6.5 um. ConrpiA solitary, acropleurogenous, ovate to obclavate, obtuse at

the apex, truncate at base, 3-4-septate, smooth, brown, apical cells very pale

brown and verruculose, 22-32 x 7-8 um.

Note: Spadicoides (Hughes 1958) is distinguished by differentiated erect,

straight or flexuous, unbranched conidiophores with polytretic, terminal and/

or intercalary conidiogenous cells that produce solitary, cylindrical, globose,

obclavate to irregular, euseptate, pigmented conidia. Thus far, 36 species have

been accepted in the genus (Xia et al. 2013). Only one species is similar to

S. sylvatica, S. yunnanensis L.G. Ma & X.G. Zhang, which differs by its conidia,

which are obpyriform to ovoid, 2-3-septate, smooth, brown, shorter, and wider

(18.5-28 x 6.5-10 tm) and have rounded or acute, hyaline, smooth apical cells

(Ma et al. 2010).

Fic. 1. Spadicoides sylvatica (holotype, XAL CB1687). a. Conidia. b. Conidiogenous cells and

conidia. c. Conidiogenous cell. Sporidesmiella mammillata (holotype, XAL CB1686). d. Conidia.

e. Conidium and conidiogenous cells. Scale bars = 10 um.

Spadicoides & Sporidesmiella spp. nov. (Mexico) ... 513

514 ... Heredia & al.

Sporidesmiella mammillata Heredia, R.F. Castafieda & R.M. Arias, sp. nov.

MycoBank MB812923 Fics 1D-E, 2D

Differs from Sporidesmiella pseudoseptata by its shorter conidia with 4-7 distosepta

(mostly 6-distoseptate) and a mammillate apical cell.

Type: Mexico, Yucatan State, Oxkutzcab municipality, Kaxil Kiuic Biocultural Reserve,

20°17’N 89°23’W, dry tropical forest, on decaying twig, 28 January 2014, coll. G. Heredia

(Holotype: XAL CB1686).

ErymMo_oey: Latin, mammillata, referring to the breast-like conidial apical cell.

Co.ontgs on the natural substrate effuse, hairy, dark brown to black. Mycelium

immersed, composed of septate, branched, brown, smooth, 1-2.5 um diam.

CONIDIOPHORES differentiated, mononematous, erect, flexuous or slightly

geniculate, with 4-10-enteroblastic percurrent extensions, dark brown or

black, paler at the end, smooth, 80-220 x 5-11 um. CONIDIOGENOUS CELLS

monoblastic, cylindrical, integrated, indeterminate, with several enteroblastic

percurrent extensions, pale brown to brown. Conidial secession schizolytic.

Conipi1a solitary, acrogenous, obclavate to fusiform, mammiform at the apex,

4-7-distoseptate (mostly 6-distoseptate), smooth, golden brown, 23-32 x

6.5-8 um.

Note: Kirk (1982) established Sporidesmiella with S. claviformis as type;

the genus is characterized by conidia that are solitary, acrogenous, mostly

distoseptate (some species with a proximal basal euseptum), pale olivaceous

brown or subhyaline and produced by a monoblastic, terminal, integrated,

indeterminate, enteroblastic percurrent elongated conidiogenous cell. In the

recent key to accepted Sporidesmiella species by Ma et al. (2012), of the six

species that can be compared with S. mammillata — S. archidendri Jian Ma &

X.G. Zhang, S. ciliaspora W.P. Wu, S. fusiformis W.P. Wu, S. machili Jian Ma &

X.G. Zhang, S. pseudoseptata (M.B. Ellis) Subram., S. verruculosa W.P. Wu —

all clearly differ (see TaBLE 1, Fic. 2).

TABLE 1. Comparison of Sporidesmiella mammillata and similar species.

SPECIES CONIDIAL SIZE (jtm) No. OF DISTOSEPTA

S. archidendri 100-160 x 13-17 19-29

S. ciliaspora 45-55 x 8-9 9-19

S. fusiformis 64-80 x 12-13 14-17

S. machili 57-110 x7.5-10 9-15

S. mammillata 23-32 x 6.5-8 4-7

S. pseudoseptata 35-56 x 7-8 5-8

S. verruculosa 85-95 x 15-21 17-18

515

Spadicoides & Sporidesmiella spp. nov. (Mexico) ...

: > mee

nase 3

ooo TOL

ati THE Eg qitas.-

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ae eae

73° @ *

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paths s.

;

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;*.

Fic. 2. Sporidesmiella representative conidia.

a. S. archidendri. b. S. ciliaspora. c. S. fusiformis. d. S. mammillata.

e. S. machili. f. S. pseudoseptata. g. S. verruculosa. Scale bars

= 20 um.

516 ... Heredia & al.

Acknowledgments

The authors express their sincere gratitude to the owners of the Kaxil Kiuic

Biocultural Reserve and to Dr. Luis EP. Gusmao and Dr. Xiu-Guo Zhang for their

critical review of the manuscript. This study was supported by the Instituto de Ecologia

A.C., Mexico. RFCR is grateful to the Cuban Ministry of Agriculture and “Programa

de Salud Animal y Vegetal” (project P131LH003033) for facilities. We acknowledge the

assistance provided by Dr. P.M. Kirk and Drs. V. Robert and G. Stegehuis through the

IndexFungorum and MycoBank websites. Dr. Lorelei Norvell’s editorial and Dr. Shaun

Pennycook’s nomenclatural reviews are greatly appreciated.

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from southern China. Mycoscience 53: 187-193. http://dx.doi.org/10.1007/S10267-011-0152-1

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and record of Pseudoacrodictys from southern China. Mycol. Progress 10: 261-265.

http://dx.doi.org/10.1007/s11557-010-0696-z

MY COTAXON

http://dx.doi.org/10.5248/130.517

Volume 130, pp. 517-532 April-June 2015

Pholiota olivaceophylla, a forgotten name for a common

snowbank fungus, and notes on Pholiota nubigena

NOAH SIEGEL’, NHU H. NGUYEN’, & ELSE C. VELLINGA?*

'25 Prospect Hill Rd, Royalston MA 01368-9206, USA

? Department of Plant Biology, 250 Biological Sciences, University of Minnesota,

1445 Gortner Ave., St. Paul, MN 55108, USA

°111 Koshland Hall #3102, University of California Berkeley, Berkeley CA 94720-3102, USA

* CORRESPONDENCE TO: ecvellinga@comcast.net

ABSTRACT — A name has been found for a common species in Pholiota subg. Flammuloides

fruiting during and soon after snowmelt in the subalpine Abies forests of California: Pholiota

olivaceophylla is characterized by rather pale slime-covered basidiocarps, relatively pale

brown ellipsoid to slightly phaseoliform spores, 6.0-8.5 x 3.5-5.0 um, with an inconspicuous

germ pore, and abundant lageniform pleurocystidia. The ITS sequence of the type collection

of Ph. olivaceophylla matches those of recent collections. From phylogenetic analyses and

morphology, it is clear that the secotioid Nivatogastrium nubigenum [=Pholiota nubigena]

is nested within Pholiota; this species has retained ballistospores and the typical curved

sterigmata for active spore dispersal.

Key worps — Abies magnifica, biodiversity, Strophariaceae

Introduction

The genus Pholiota (Fr.) P. Kumm. is generally characterized by (pale) yellow

to brown basidiocarps with (in most species, notably excepting the type) a

viscid to gelatinous, often scaly, pileus, an annulus, rusty to dark brown smooth

to slightly rough spores with a germ pore, cheilocystidia and pleurocystidia

(in a number of species as chrysocystidia), and typically lignicolous habit (e.g.,

Jacobsson 2009). Smith & Hesler (1968), in their overview of North American

species of Pholiota, used a rather broad genus concept but a narrow species

concept and gave descriptions of 205 species for North America, 163 of which

were described as new. Subsequently, several of these species were shown to

be synonyms of each other or to belong to other genera that even Smith &

Hesler (1968) recognized as distinct, e.g., Agaricus L. and Stropharia (Fr.) Quel.

518 ... Siegel, Nguyen, & Vellinga

(Jacobsson 1990, 1997; Noordeloos 1999; Norvell & Redhead 2000; Redhead

1984a,b). In modern treatments of Pholiota for various regions in Europe with

narrower genus concepts, 24-28 species are recognized (Holec 2001, Jacobsson

2009, Noordeloos 2011, Holec & Kolarik 2014, Holec et al. 2014) [excluding,

among others, Crassisporium Matheny et al., Flammula' (Fr.) P. Kumm.,

Flammulaster Earle, Hemipholiota (Singer) Bon, Hemistropharia Jacobsson &

E. Larss., Kuehneromyces Singer & A.H. Sm., Meottomyces Vizzini, Phaeolepiota

Maire ex Konrad & Maubl., Phaeonematoloma (Singer) Bon, Phaeomarasmius

Scherff., and a number of Tubaria species, comprising 40-50 species]. The

reasons for the discrepancy in species totals between Europe and North America

are multiple; it is due partly to a strong difference in both genus and species

concepts and reliance on poorly documented earlier descriptions in order to be

comprehensive or partly reflects variation in fresh basidiocarp characteristics

leading to multiple descriptions of species. The aspect of the basidiocarps

changes dramatically due to weather - the slimy species are dry during dry

weather; furthermore, in the species of Pholiota subg. Flammuloides A.H. Sm.

& Hesler the pleurocystidia show considerable variation in incrustation, wall

thickness, and coloration among collections of the same species (see below).

As a result species recognition based on morphology alone can be challenging.

Many new species described by Smith & Hesler (1968) were based on only

one collection, and the authors probably underestimated the variability of the

species they described.

A common snowbank species in Pholiota subg. Flammuloides (with a

gelatinous pileus surface and conspicuous lageniform pleurocystidia but

without chrysocystidia) was regularly found in Californias Red Fir (Abies

magnifica A. Murray bis) zone in the Sierra Nevada and Southern Cascade

mountains. For a number of years the fungus was considered undescribed

(Arora 1986), but a scrupulous combing of Smith & Hesler’s 1968 Pholiota

monograph turned up one possible name: Pholiota olivaceophylla. This species

was described based on one collection, occurring ‘on and near very rotten

Shasta fir log, Mt. Shasta, Calif. June 1954’ (Shasta fir is the vernacular name for

A. magnifica var. shastensis Lemmon). Besides being collected at the right time

and place, the morphology also fits our unnamed species, except for the color

of the lamellae. We never found olivaceous tinges in the (young) lamellae of

'The name Flammula (Fr.) P. Kumm., published in 1871, is a later homonym of Flammula

(Webb ex Spach) Fourr. from 1868, a genus in the Ranunculaceae; Donk (1962) considered

the latter to be invalid, but his point of view is not followed by later authors such as

Jacobsson (2012). A proposal for conservation of the fungal genus Flammula has been put

forward by Redhead (2013).

Pholiota olivaceophylla & P. nubigena (U.S.A.) ... 519

our specimens (PLATE 2). Subsequently, the microscopic features of the type of

Ph. olivaceophylla were examined and the ITS sequences of the type and recent

collections were compared. Here, based on a wide sampling from different

locations within California, we present a thorough description of the species

with a photograph, and we place the ITS sequences in the context of the whole

genus to evaluate the taxonomic position of the species.

Pholiota olivaceophylla is considered a snowbank fungus as described by

Cooke (1944, 1955). Snowbank fungi depend on the moisture provided by the

melting snow for the development of their basidiocarps, and many are able

to fruit under the snow. Pholiota olivaceophylla starts fruiting in this way, but

continues fruiting after snow melt. The fungus long known as Nivatogastrium

nubigenum is another example of this ecological group. The snowbank fruiting

habit seems to be restricted to the western North American mountains and has

not been encountered in Europe. Cripps (2009) gave an overview of the more

common snowbank species; with the description of Baeospora occidentalis

L.J. Hutchison & Kropp from Utah (Hutchison et al. 2012), the number of

known snowbank fungi has already increased.

Materials & methods

MorpHOLoGy—Standard methods for describing basidiocarps were applied, using the

terminology of Vellinga & Noordeloos (1999). Color annotations in the macroscopical

descriptions follow Kornerup & Wanscher (1967). Microscopical observations were

made from dried material rehydrated in 10% ammonia. The notation [81, 4, 3] indicates

that measurements were made on 81 spores in four samples in three collections; spores

were observed and measured from deposits on the stipe apex, and at least 20 spores

were measured per collection in side view; also for a few collections, at least 20 spores

per collection were measured in frontal view. The following abbreviations are used:

L = number of lamellae, 1 = number of lamellulae between two lamellae, avl = average

length, avw = average width, Q = quotient of length and width, and avQ = average

quotient. The genus name Pholiota is abbreviated as Ph. Collections are in UC, except

the type collection (in MICH). Herbarium abbreviations are according to Thiers (2015,

continuously updated). [For full collection information (including Genbank accession

numbers) see http://mycotaxon.com/Vvol/130/Siegel_Table1.pdf]

DNA EXTRACTION & SEQUENCING—Collections for this study were made specifically

during the Yosemite National Park fungus survey, and during the National Forest

Service surveys in the southern Cascade Range in California. We extracted DNA using a

modified Sigma Extract-N-Amp kit (Sigma Aldrich, St. Louis, MO) for dried mushrooms

and Whatman™ FTA™ Plant cards (Whatman Ltd) for dried tissue embedded in the

cards following manufacturer’s instructions. The nrITS region was amplified with the

ITS1F/ITS4 primer set using an MJ PTC-100™ thermocycler (Applied Biosystems, Foster

City, CA, USA) under conditions previously described (Gardes & Bruns 1993). PCR

520 ... Siegel, Nguyen, & Vellinga

products were cleaned using 0.5 uL of ExoSAP IT (USB Corp, Cleveland, OH, USA) per

reaction and cycled at 37°C for 45 min, followed by 80°C for 15 min. Sequencing was

performed using Big Dye chemistry and an ABI PRISM 3700 Genetic Analyzer (both

from Applied Biosystems, Foster City, CA, USA). Sequences were edited and contigs

were assembled using Sequencher 4.7 (Gene Codes Corporation, Ann Arbor, MI, USA),

and deposited in GenBank.

ALIGNMENT & PHYLOGENETIC ANALYSES— The nrITS sequences newly produced for this

paper, plus all nrITS1-5.8S-ITS2 sequences of Pholiota, Flammula, and Nivatogastrium

species and their environmental counterparts from GenBank (as of October 2013), were

aligned with the program MAFFT version 7 (Katoh et al. 2002), and visually optimized.

Two sequences in GenBank, GU234152 and GU234164, were excluded as they show

92% BLAST similarity with a Conocybe teneroides collection (JF08598), and are not

close to Pholiota. The aligned sequence database was analyzed by maximum likelihood

method (ML) using the on-line program RAXML version 7.7.1 (Stamatakis et al. 2008);

100 rapid ML bootstraps were performed, and bootstrap values are included in the

ML tree of PLaTE 1. Pholiota lignicola [= Kuehneromyces lignicola (Peck) Redhead] and

Flammula sp. (coll. UC1861000) were chosen as outgroup taxa.

Results

The ITS alignment consists of 131 sequences (25 newly produced and 22

from California collections) and is 801 base pairs long. The resulting maximum

likelihood tree shows that Pholiota olivaceophylla is well supported (100%)

(PLATE 1; see http://mycotaxon.com/vol/130/Siegel_Platel.pdf for a larger

online version). Its closest relatives in our analyses are a collection from

Scandinavia labeled Ph. “spumosa” and the isotype of Ph. ferrugineolutescens

A.H. Sm. & Hesler from northern coastal California. Pholiota olivaceophylla

belongs to a clade that corresponds with the morphologically delineated

subg. Flammuloides to which other common Californian taxa, such as Ph.

velaglutinosa A.H. Sm. & Hesler, Ph. highlandensis (Peck) Quadr. & Lunghini,

and Ph. spumosa (Fr. : Fr.) Singer belong. Nivatogastrium nubigenum, which

forms secotioid basidiocarps, falls in the middle of subg. Flammuloides, close to

a collection of an agaricoid Pholiota from Yosemite National Park and Pholiota

sp. FJ717493 from Canada. Flammula is separate from Pholiota, as already

shown by Gulden et al. (2005), and Matheny et al. (2015) in their work on

various brown-spored groups, and in more wide-ranging works on Agaricales

as a whole (e.g., Matheny et al. 2007, Moncalvo et al. 2002). Subgenera Pholiota

and Flammuloides are clearly separate as well (PLATE 1).

Discussion

The coverage of the genus Pholiota in terms of ITS sequences is far from

complete; some groups are well represented, but as a whole there is obviously

much work to be done at all taxonomic levels. The goal of our phylogenetic

100 Pholiota lucifera Korea

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oliota multicingulata

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oliota mixta UC1

HG007988 Fholiota gallica holotype MPU EUR

100 8 F908583 Pholiota chocenensis MCVE112

G007985 Pholiota chocenensis holotype

KC176331 environmental sample MI

HQ222025 Pholiota castanea type TENN020269 TN

86 Pholiota mixta Mex

holiota carbonaria E

98 UC1999437 CA

Pholiota carbonaria Korea

Pholiota highlandensis PRM888152 EUR

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Pholiota highlandensis EUR

Pholiota highlandensis PRM887.

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oliota nameko Korea

oliota nameko China.

Pholiota adiposa China

Pholiota gummosa MCVE65.

Pholiota caespitosa holotype

Pholiota olivaceodisca ho’ ety

Pholiota tennesseensis holo

Pholiota burkei holotype TE

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environmental sample EUR

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PiaTE 1. ITS maximum likelihood tree of the genus Pholiota. Numbers on branches are ML

bootstrap values >69. Each terminal leaf

is annotated as: GenBank accession; Taxon name;

Herbarium accession; Location. Location abbreviations: AK = Alaska, AL = Alabama, BC = British

Columbia (Canada), CA = California, EUR = Europe, Mex

Mexico, MI = Michigan,

NC = North Carolina, NZ = New Zealand, Ont = Ontario (Canada), TN = Tennessee. Newly

produced sequences are in bold.

522 ... Siegel, Nguyen, & Vellinga

analysis was not to deliver a thorough hypothesis on the evolution and

phylogenetic relationships of Pholiota taxa, but to show the grouping and

position of Ph. olivaceophylla and Ph. nubigena in general terms. As is clear

from PLATE 1, species concepts are in need of reconsideration, as identical

sequences have been produced from collections that had been identified as

different species (e.g., in the Ph. adiposa clade), and the same name had been

applied to collections that belong to different clades (e.g., Ph. spumosa).

Sequencing of type collections, as started by Matheny and co-workers (in

TENN), should be expanded to all species described by Smith & Hesler (1968),

and neotypes and epitypes should be selected and presented with their sequence

data. Holec et al. (2014) also suggested increased sampling and sequencing

efforts to better understand the diversity and variability of the species.

Striking is that Pholiota castanea A.H. Sm. & Hesler (ITS GenBank

HQ222025 from the type collection), described as the only other species in

stirps Olivaceophyllae, from the Great Smoky Mountains National Park, is

not closely related to Ph. olivaceophylla (PLATE 1), but is close to the recently

described European Ph. chocenensis Holec & M. Kolarik. Pholiota castanea and

Ph. olivaceophylla stand out among the species in subg. Flammuloides because

of their non-gelatinized subhymenium (Smith & Hesler 1968).

Based on an analysis of a 6-locus database of the Agaricales as a whole,

the secotioid N. nubigenum is a sister taxon to Pholiota squarrosa (Vahl : Fr.)

P. Kumm., the only agaricoid Pholiota species included in that analysis (Matheny

et al. 2007); it is a sister taxon to Pholiota multicingulata E. Horak in another

study of the Agaricales based on a 4-locus database (Matheny et al. 2015). In

the present ITS analysis, Ph. nubigena (as Nivatogastrium nubigenum in PLATE

1) clearly nests within Pholiota subg. Flammuloides. Redhead (2014) proposed

the new combination of this species in Pholiota.

Taxonomy

Pholiota olivaceophylla A.H. Sm. & Hesler, N. Am. Species Pholiota: 236. 1968.

PLATES 2-6

Piteus (30-)40-80(-100) mm, convex to broadly convex, becoming

applanate, occasionally wavy or lobed in age; margin inrolled when young,

becoming even, undulating in age, often with veil remnants when young; surface

smooth, viscid when wet, becoming tacky, often with debris stuck to it, shiny

when dry, creamy-orange, pale orange-buff to pale orange-tan when young,

becoming creamy-buff to tan, often with darker orange-tan streaks or patches;

slime pale orange. LAMELLAE, L = about 50-60, | = 1-3, close to subdistant,

adnate with decurrent tooth, 7-10 mm wide, pale tan to buff (5B3-5C4) when

young, becoming tan, light brown to pale olive-brown, with even concolorous

Pholiota olivaceophylla & P. nubigena (US.A.) ... 523

PLATE 2. Pholiota olivaceophylla (UC1999285). Photo by Noah Siegel

edges. STIPE 50-90 X 10-18 mm, equal or enlarged at base, whitish when

young, covered with pale orange-buff slimy veil except for apex, becoming off-

white to dingy tan with pale orange streaks, dry and white above veil, becoming

dingy tan in age, at base covered with fuzzy white mycelium that binds to

organic matter, stuffed when young, becoming hollow in age. PARTIAL VEIL a

mucilaginous cortina, breaking up and leaving remnants around pileus margin

and a faint very pale orange ring that becomes brown with spores. CONTEXT

white to pale cream in pileus, watery-white to pale tan in stipe, becoming tan to

pale brown. Opor musty, herbaceous. Taste mild. CHEMICAL REACTIONS: 5%

KOH on pileus pale yellowish orange, on upper stipe orange, little reaction on

lower stipe, on pileus context bright orange, becoming orange-brown, on stipe

context orangish. SPORE PRINT color medium brown.

BASIDIOSPORES in side view [200, 10, 10] 5.9-8.3 x 3.5-4.4 um, avl x avw

= 64-7.3 X 3.9-4.1 um, Q = 1.45-1.95, ellipsoid to oblong, some slightly

phaseoliform, in frontal view [40, 2, 2] 6.0-7.6 x 3.4-4.4 um, avl x avw = 6.4-6.8

x 4.0-4.2 um, Q = 1.4-1.9, avQ = 1.55-1.8, relatively thin-walled; germ pore

and hilar appendage inconspicuous. BASIDIA 17-29 X 5-8 um, 4-spored, with

basal clamp connection. LAMELLA EDGE Sterile. CHEILOCYSTIDIA 25-65 X 8-15

um, with 4-6 um wide neck, lageniform, utriform, or clavate, a few similar in

shape and size to pleurocystidia, variously incrusted, with coarse material or

524 ... Siegel, Nguyen, & Vellinga

PiaTE 3. Pholiota olivaceophylla. a. Cheilocystidia; b. pleurocystidia (both from UC UC1999268).

Scale bars = 10 um. Drawing by ECV.

with some small pieces, in some collections with yellow contents in ammonia.

PLEUROCYSTIDIA 48-91 X 10-17.5 um, with 4-10 um wide neck, lageniform,

very abundant, protruding from hymenium, variable in size and ornamentation,

yellow in ammonia in some collections, with or without an apical covering

of crystalline material, or with amorphous matter inside, or without any

inclusions, with slightly thickened walls in lower part of neck. HYMENOPHORAL

TRAMA not pigmented. SUBHYMENIUM not gelatinized. PILEIPELLIS an ixocutis

of cylindrical non-colored hyphae, 2-5 um wide; subpellis a cutis of cylindrical

to slightly inflated hyphae, 5-10 um wide with brown walls. STIPITIPELLIS a

cutis of cylindrical colourless hyphae, 4-11 um wide, without caulocystidia.

CLAMP CONNECTIONS present in all tissues.

SPECIMENS EXAMINED: USA: CALIFORNIA, Siskiyou Co., Modoc National Forest,

north slope of Black Mountain, near junction of Forest Road 44N33 & 44N12, in Abies

magnifica & A. concolor forest, 29 May 2012, E.C. Vellinga (UC 1999279); in A. magnifica

& A. concolor forest on small branches and thick duff near melting snow, 29 May 2012,

N. Siegel (UC 1999281; GenBank KC122883); in A. magnifica & A. concolor forest on

conifer wood, 29 May 2012, E.C. Vellinga (UC 1999285; GenBank KC122888); in A.

magnifica & A. concolor forest on conifer wood & twigs, 29 May 2012, E.C. Vellinga (UC

1999276); in A. magnifica & A. concolor forest, 29 May 2012, N. Siegel (UC 1999280);

Pholiota olivaceophylla & P. nubigena (U.S.A.) ... 525

g.9 6 Q 6

PLATE 4. Pholiota olivaceophylla. a. Spores; b. basidia; c. cheilocystidia; d. pleurocystidia

(all from UC1999272). Scale bars = 10 um. Drawing by ECV.

Mount Shasta, on and under rotten A. magnifica log, 1 June 1954, W.B. Cooke (MICH

290502, holotype; GenBank KF878381); Mount Shasta, near Clear Creek Trailhead, in

A. magnifica forest, 28 May 2012, E.C. Vellinga (UC 1999277); Trout Creek area, in A.

magnifica and A. concolor forest, on conifer wood and twigs, 28 May 2012, E.C. Vellinga

(UC 1999283; GenBank KC122889); Six Shooter Butte, in A. magnifica forest, on conifer

wood, 27 May 2012, E.C. Vellinga (UC 1999278); Shasta National Forest, Trout Butte

area, 28 May 2012, N. Siegel (UC 1999284; GenBank KC122878); Tuolumne Co.,

Yosemite National Park, Crane Flat, north of gas station, in mixed coniferous forest, on

conifer wood, 4 June 2010, N. Siegel (UC 1999268; GenBank KC122882); Tamarack Flat

Road near Tioga Road, 37.7966°N 119.8655°W, in A. magnifica forest, on conifer wood,

5 June 2010, D. Viess (UC 1999272; GenBank KC122885); Mariposa Co., Yosemite

National Park, Glacier Point Road, 37.67333°N 119.6523°W, in mixed coniferous forest,

30 May 2010, M. Schubert (UC 1998508; GenBank KC122894); Glacier Point Road,

Summit Meadow, 37.6650°N 119.6997°W, in P jeffreyi, P. lambertiana, & Abies forest, on

litter, 30 May 2012, N.H. Nguyen (UC 1998643; GenBank KC122895).

HABITAT & DISTRIBUTION—Solitary, scattered, or in small groups and clusters

on small buried branches, woody debris, thick duff, or well rotted logs, in Abies

concolor (Gordon) Lindl. ex Hildebr. and A. magnifica forests at 1700-2200 m

asl, fruiting near melting snowbanks or areas with snow cover, occasionally

fruiting under snow, May-June. Widespread in the Sierra Nevada and southern

Cascades in California.

526 ... Siegel, Nguyen, & Vellinga

QO@QQAVGDO

b Cc

Pate 5. Pholiota olivaceophylla. a. Spores; b. cheilocystidia; c. pleurocystidia

(all from UC1998643). Scale bars = 10 um. Drawing by ECV.

The type collection was in good condition, and the dried specimens of recent

collections look very similar. The following characters were noted for the type

collection (PLATE 6):

SPorEs [20, 1] 6.4-7.8 x 3.4-4.7 um in side-view, avl x avw = 7.0 X 3.9 um,

Q = 1.67-1.89, avQ = 1.8, elongate, slightly phaseoliform, pale brown, with

inconspicuous germ pore and hilar appendage. Basip1a 20-23 x 5.0-6.5 um,

4-spored, with basal clamp connection. CHEILOCysTIDIA present, difficult

to observe, lageniform. PLEUROCysTIDIA abundant, 63-80 x 11-16 um,

lageniform, with 15-35 um long cylindrical, rarely ventricose neck, often

slightly pinched in near the wider part, with or without yellowish contents,

slightly thick-walled.

ADDITIONAL SPECIMENS EXAMINED:

Flammula sp.: USA: CALIFORNIA, Mariposa Co., Yosemite National Park, White Wolf

campground, 37.869383°N -119.646585°W, in A. concolor and P. contorta forest, on

conifer wood, 18 Aug. 2010, P.G. Werner (UC 1860901; Genbank KC122893); Yosemite

National Park, Glacier Point Road, 37.7065°N -119.5890°W, in Pinus contorta forest, 29

Oct. 2011, L. Rosenthal (UC 1861000; Genbank KC122880).

Pholiota cf. brunnescens: USA: CALIFORNIA, Mariposa Co., Yosemite National Park,

Mariposa Grove loop trail, 37.50568°N -119.60847°W, in mixed coniferous forest,

Pholiota olivaceophylla & P. nubigena (U.S.A.) ... 527

cs lihh

Pate 6. Pholiota olivaceophylla. a. Spores; b. basidia; c. cheilocystidium; d. pleurocystidia

(all from holotype). Scale bars = 10 um. Drawing by ECV.

29 May 2010, N.H. Nguyen (UC 1998624; Genbank KC122891); Tuolumne Co.,

Yosemite National Park, Tamarack Flat Road, 37.7966°N -119.8655°W, in PB. ponderosa,

Pseudotsuga menziesii & Calocedrus decurrens forest, N. Siegel, 5 June 2010 (UC

1999269; Genbank KC122892).

Pholiota cf. decorata: USA: ALAsKa, Anchorage Co., Girdwood, Iditarod Trail, in

forest with Picea sitchensis and Tsuga, 31 Aug. 2012, E.C. Vellinga 5567b (UC 1999442;

Genbank KF878380).

Pholiota highlandensis: USA: CALIFORNIA, Mendocino Co., Mendocino, off Wheeler

Street, on burnt ground under Pinus, 2 Jan. 2013, N. Siegel (UC 1999437; Genbank

KF878376).

Pholiota lenta s.l.: USA: CALIFORNIA, Yuba Co., Bullard’s Bar Recreation Area,

Hornswaggle Campground, 39.4146°N -121.1205°W, mixed conifer forest, on

coniferous wood, 10 Dec. 2011, E.C. Vellinga (UC 1999282; Genbank KC122876).

Pholiota mixta: USA: ALASKA, Kenai Peninsula, Hope, along Palmer Creek Road, 3

Sept. 2012, E.C. Vellinga 5580b (UC 1999441; Genbank KF878377).

Pholiota sp.: USA: ALASKA, Kenai Peninsula, Hope, along Palmer Creek Road, 3 Sept.

2012, E.C. Vellinga 5578b (UC 1999439; Genbank KF878378); CALIFORNIA, Yosemite

National Park, Highway 120, in A. magnifica forest, on conifer wood, 30 Sept. 2011,

D. Rust (UC 1999274; Genbank KC122881); NoRTH CAROLINA, Mount Mitchell State

Park, on conifer wood, 22 Sept. 2012, D. Viess (UC UC 1999438; Genbank KF878379).

528 ... Siegel, Nguyen, & Vellinga

Pholiota spumosa: USA: CALIFORNIA, Alameda Co., Berkeley, along Euclid Avenue,

on Pinus bark chips in garden, 18 March 2012, E.C. Vellinga (UC 1999286; Genbank

KC122886); Mariposa Co., Yosemite National Park, Wawona Meadow Loop Trail,

37.523056°N -119.636111°W, on buried conifer wood, 29 May 2010, E.P. Blanchard

(UC UC 1998527; Genbank KC122879).

Pholiota terrestris: USA: CALIFORNIA, Marin Co., Point Reyes National Seashore,

Olema Trail, 37.9917°N -122.7580°W, in Ps. menziesii & hardwood forest, 9 Dec.

2006, K. Takeoka (UC 1859859; Genbank KC122896); Point Reyes National Seashore,

Bayview Trail, 38.0608°N -122.8564°W, in P. muricata forest, 29 Dec. 2007, Aletheap

(UC 1860187; Genbank KC122887).

Pholiota velaglutinosa: USA: CALIFORNIA, Marin Co., Point Reyes National Seashore,

Mount Vision, 38.1016°N -122.8947°W, in P. muricata forest, 10 Dec. 2005, Murray (UC

1859567; Genbank KC122877).

ComMMENTS— The data noted from the type collection concur with those given

by Smith & Hesler (1968) and fit well the modern data for the collections from

California's mountain ranges. An image of A.H. Smith's original notes can be

found in Mycoportal (http://mycoportal.org).

Pholiota olivaceophylla commonly fruits in May and June during and just

after snowmelt in the Abies forests of the Sierra Nevada and Cascade ranges.

It can easily be distinguished from the other common species in Pholiota

subg. Flammuloides fruiting at the same time in the mountains. For example,

Ph. highlandensis and Ph. brunnescens A.H. Sm. & Hesler grow on recent burns,

have smaller basidiocarps and deeper brown, thick-walled spores. Species in

the Ph. spumosa complex enjoy bright yellow colors in the basidiocarps, and

have brown thick-walled spores with a distinct germ pore, which are similar in

size to those of Ph. olivaceophylla.

The European species Ph. elegans Jacobsson comes close in overall

appearance, but differs in habitat (mainly in virgin, often deciduous, forests and

fruiting in autumn), and slightly smaller, distinctly phaseoliform-amygdaloid

spores, 5.0-6.7(—7.0) um long (Holec 2001).

The North American species in Pholiota subg. Flammuloides are in serious

need of revision, and interpretation of many species described by Smith and

Hesler (1968) is challenging. However, some species yield to interpretation and

appear to be close to Ph. olivaceophylla in their pale colors, but differ in critical

characters: the pale species Ph. flavopallida A.H. Sm. & Hesler has spores 5-6

uum long, and Ph. agglutinata A.H. Sm. & Hesler has a white veil, and was found

in late summer. There are, as far as we know, no other spring-fruiting mountain-

dwelling species described. McClenaghan (1991) in her overview of northern

California species gave a relatively short description of Ph. olivaceophylla based

on one collection (which we have not studied), and stressed the olivaceous

lamellae that turn to dark brown with age, characters we did not observe in the

Pholiota olivaceophylla & P. nubigena (U.S.A.) ... 529

many specimens we collected. Other characters (including the location and the

time of collecting) do fit with our concept of Ph. olivaceophylla.

The pleurocystidia of Ph. olivaceophylla are highly variable in size, in

contents, and in ornamentation among collections (PLATES 3-6); how this is

influenced by age of the basidiocarps and environmental variables is unknown.

There is no overlap in pleurocystidium size among the collections with the

shortest and those with the longest pleurocystidia. Constant are the relatively

thin walls and relatively pale brown color of the spores under the microscope,

and inconspicuous germ pore and hilar appendage. Spore color is a much

better character at species level than pleurocystidium shape and size, but it is a

qualitative character, and thus harder to assess.

Pholiota nubigena (Harkn.) Redhead, Index Fungorum 148: 1. 2014. PLATE 7

= Secotium nubigenum Harkn., Bull. California Acad. Sci. 1(4): 257. 1886.

= Nivatogastrium nubigenum (Harkn.) Singer & A.H. Sm., Brittonia 11: 224. 1959.

CoMMENTS—Pholiota nubigena shares both the hallmarks for subg.

Flammuloides and the habitat with Ph. olivaceophylla. The pleurocystidia are

: OO;

Pate 7. Pholiota nubigena. a. Spores; b. basidia; c. pleurocystidia (all from UC1998744).

Scale bars = 10 um (note different scale for c than in the other figs.). Drawing by ECV.

530 ... Siegel, Nguyen, & Vellinga

numerous and lageniform with a very long neck without yellow inclusions

(chrysocystidia are absent), the spores are larger than those of Ph. olivaceophylla

(based on our observations on collection UC 1998744 [25, 1, 1] 7.8-10.8 x

4,9-6.2 um in side-view), with a medium brown, medium thick wall, a distinct

hilar appendage, and a hardly visible germ pore (PLATE 7a).

The spores are asymmetrical in side view, and the basidia have sterigmata

typical for ballistospore dispersal (PLATE 7b). Pholiota nubigena grows, like

Ph. olivaceophylla, on conifer wood, and is widespread in the western North

American mountains, with records from the southern Sierras northwards into

Idaho and the Rocky Mountains in Montana (http://mycoportal.org retrieved

12 February 2015).

SPECIMEN EXAMINED: USA: CALIFORNIA, Mariposa Co., Yosemite NP, Glacier Point

Road, 37.6687°N 119.6113°W, M.G. Wood, 30 May 2010, M.G. Wood (UC 1998744).

Phylogenetically Ph. nubigena falls in the middle of subg. Flammuloides, and a

relationship with Pholiota — which has been proposed in the past, both prior to

(e.g. Singer & Smith 1959, Smith & Hesler 1968) and subsequent to molecular

analyses (Matheny et al. 2007, 2015) — is again confirmed here.

Acknowledgments

Matthew Foltz and Patricia Rogers at MICH were so kind as to arrange the loan

of the type of Ph. olivaceophylla. Alex Grote and Sara Zlotnik helped produce some of

the DNA sequences. This work was partially funded by the Shasta-Trinity and Lassen

National Forest as part of Northwest Forest Plan strategic surveys. Comments by the

reviewers, Drs Scott A. Redhead and Joe F. Ammirati, and by the nomenclature editor

Dr. Shaun Pennycook, on earlier versions are highly appreciated.

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MY COTAXON

http://dx.doi.org/10.5248/130.533

Volume 130, pp. 533-541 April-June 2015

Leucoagaricus lahorensis,

a new species of L. sect. Rubrotincti

T. Qasim”, T. Amir, R. NAWAZ, A.R. NIAZI, & A.N. KHALID

Department of Botany, University of the Punjab,

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

*CORRESPONDENCE TO: tayyaba.qasim@yahoo.com

ABSTRACT—Leucoagaricus lahorensis sp. nov. is described and illustrated based on morpho-

anatomical and molecular (ITS) characteristics. The new species, which is placed in L. sect.

Rubrotincti, is characterized by an umbonate to plane pileus with central obtuse orange red

disc, a white to cream stipe, ascending annulus, broadly ellipsoid basidiospores, narrowly

clavate to subcylindrical cheilocystidia without crystals at their apex, and straight cylindrical

pileal elements.

KEY worDs—Agaricaceae, macrofungi, phylogenetic analysis, Punjab

Introduction

Leucoagaricus Singer (Agaricaceae, Agaricales) is represented by more than

100 species worldwide (Kirk et al. 2008, Kumar & Manimohan 2009, Liang et

al. 2010, Vellinga et al. 2010, Munoz et al. 2012 & 2013, Malysheva et al. 2013).

The genus is characterized by non-striate pileus margins, free lamellae, spores

that are metachromatic in cresyl blue, and the absence of clamp connections

and pseudoparaphyses (Singer 1986, Vellinga 2001). Leucoagaricus has been

shown to be polyphyletic (Vellinga 2004).

Most Leucoagaricus species have been described from temperate regions in

North America and Europe, and little is known about the genus from tropical

areas (Liang et al. 2010). Representatives of L. sec. Rubrotincti form moderately

fleshy basidiomata with a context that does not change color when damaged

and usually a red brown, pinkish ochre, olive, or orange pileus with radially

arranged hyphae (Singer 1986). Basidiospores are characterized by a smooth

surface and incomplete or absent germ pores.

Lepiotaceous fungi occur widely in Punjab at lower elevations. Previously

only two Leucoagaricus species, L. leucothites (Vittad.) Wasser and L. serenus

534 ... Qasim & al.

(Fr.) Bon & Boiffard, have been reported from Pakistan (Ahmad et al. 1997).

During macrofungal surveys conducted in the monsoon seasons of 2011-13,

a species was encountered that differs macro- and microscopically from other

species of Leucoagaricus. A detailed description and illustrations of this novel

species are presented.

Materials & methods

Morpho-anatomical analysis

Basidiomata of Leucoagaricus lahorensis were carefully collected, photographed, and

vouchered. The dried specimens were characterized morphologically using terminology

of Vellinga & Noordeloos (2001). Colours were designated according to Munsell (1975).

Basidiocarps were sectioned and placed in 5% KOH and stained with phloxine for

microscopic observation. Colour reactions were also noted in Melzer’s reagent. At least

eighty basidiospores from two collections and twenty each of basidia, cheilocystidia,

and pileal and stipe elements were measured using an ocular micrometer. The following

abbreviations are used: avl = average length, avw = average width, Q = length/width of

basidiospores, avQ = the mean Q of all basidiospores. Drawings of these microscopic

features were made using a camera lucida attached to a compound microscope. The

holotype and paratype specimens have been deposited in the Herbarium, Department

of Botany, University of the Punjab, Lahore, Pakistan (LAH).

DNA extraction, PCR amplification, & DNA sequencing

DNA was extracted by the CTAB method (Bruns 1995). The internal transcribed

spacer region (ITS) was amplified using universal primers ITS4 and ITS1F (Gardes

& Bruns 1993). PCR conditions were as described in Gardes & Bruns (1993). The

amplified products were directly sequenced using the same pair of universal primers

(Macrogen, Korea). The nucleotide sequence comparisons were performed through

Basic Local Alignment Search Tool (BLAST) network services using National Centre

for Biotechnology Information (NCBI), USA database. For phylogenetic analysis and

sequence alignment, sequences of closely related species were retrieved from GenBank.

Sequence alignment was done in Muscle Multiple sequence alignment and phylogenetic

analysis was performed using Molecular Evolutionary Genetic Analysis (MEGA 5)

software. Phylogenetic analyses were made by Maximum Likelihood method using

Jukes Cantor Model (Tamura et al. 2011). The nucleotide sequences were submitted to

GenBank.

Results

Leucoagaricus lahorensis Qasim, Amir & Nawaz, sp. nov. FIGs 1, 2

MycoBank MB808587

Differs from Leucoagaricus rubrotinctus by its thin-walled larger broadly ellipsoid to

amygdaloid spores and narrowly subclavate to sub-cylindrical cheilocystidia.

Type: Pakistan, Punjab, Lahore, grounds of University of the Punjab, 217 m a.s.l.,

gregarious on soil, 3 September 2012, A.N. Khalid T25 (Holotype, LAH 10042012;

GenBank KJ701794).

Leucoagaricus lahorensis sp. nov. (Pakistan) ... 535

LIN

Fic. 1: Leucoagaricus lahorensis: basidiomata A-E (A & D-E, Holotype, B, T38; C, CM36) in their

natural habitat. Scale bar: A = 13 mm, B & C= 18 mm, D-E = 20 mm.

536 ... Qasim & al.

Fic. 2: Leucoagaricus lahorensis (Holotype):

A, basidiospores; B, basidia; C, cheilocystidia; D, stipe covering; E, pileus covering.

Scale bar: A-B = 6 um, C = 9 um, D = 47 um, E = 30 um.

Erymo oey: from the type locality Lahore, provincial capital of Punjab, Pakistan.

BASIDIOMATA 3.8-7.3 cm tall. PILEUS 2.5-5.5 cm in diameter, at first umbonate,

then becoming plane with deflexed margins at maturity; central disc obtuse, dark

reddish brown (10 R 3/10); surface squamulose, with radially arranged reddish

brown fibrils, becoming sparse and light brown (7.5R5/16) towards margins

over a white to cream background; context white, fleshy; margins entire, with

cracks developing at maturity. LAMELLAE free, close, thin, membranous, white

to cream (SYR9/2), edges entire; lamellulae in 3-4 tiers. STIPE 2.9-5.6 x 0.3-0.7

Leucoagaricus lahorensis sp. nov. (Pakistan) ... 537

cm, central, equal to sub-equal, cylindrical, slightly attenuating towards pileus,

with shiny white surface (S5YR9/2), hollow, smooth; base slightly bulbous.

ANNULUS present, membranous, white to cream, ascending, superior. VOLVA

absent. TASTE and opourR not recorded.

BasIpD1osPporEs [80/4/4], (7.3)8-10.6(10.9) x (6.1)6.4-7.6(7.8) tum, avl x avw =

8.7 x 6.7 um, Q = 1.2-1.4, avQ = 1.3, broadly ellipsoid to ellipsoid, occasionally

amygdaloid, hyaline with greenish tinge in KOH, thin-walled, smooth,

apiculate, guttulate, dextrinoid in Melzer’s reagent; germ pore absent. BASIDIA

(17.2)17.5-18.9(19.1) x (10.1)10.6-11.6(11.8) um, avl x avw = 17.8 x 11.2 um,

thin-walled, hyaline in KOH, 2-4-spored, broadly clavate to vesicular, smooth.

CHEILOCYSTIDIA (33.7)34.1-38.7(40) x (8)9.1-10.5(11.3) um, avl x avw =

24.2 x 10.7 um, hyaline in KOH, thin-walled, narrowly clavate to sub-

cylindrical, smooth. PLEUROCYSTIDIA not present. STIPITIPELLIS made of cells

11.8 - 23.6 um diam., avw = 17.6 um, thin-walled, hyaline in KOH, parallel

in arrangement, septate, unbranched; clamp connections absent. PILEIPELLIS

hyphae 5.7-11.6 um (av = 7.5 um) diam, light brown in colour. Clamp

connections absent.

ADDITIONAL MATERIAL EXAMINED: PAKISTAN, Punjas, Lahore, University of the

Punjab, 217 maz.s.l., gregarious on rich loamy soil, 18 August 2011, A.N. Khalid MCR84

(LAH 8482011; GenBank KJ701797); on grounds of botanical garden, 21 September

2012, A.N. Khalid & Tayyaba Qasim T38 (LAH 10052012; GenBank KJ701794); Changa

Manga Forest, on rich soil, 28 July 2013, A.R. Niazi & Tooba Amir CM36 (LAH 2481336;

GenBank KJ701795).

Phylogenetic analysis

ITS-nrDNA amplification produced fragments of 700 -742 bp. The initial

BLAST analysis revealed the maximum similarity of L. lahorensis with

L. rubrotinctus. The top 100 sequences in BLAST analysis that represent

Leucoagaricus and other related taxa were retrieved for phylogenetic analysis.

From the final data set comprising 48 sequences, 47 represented the ingroup,

while Cystolepiota seminuda (AY 176350) was selected to root the tree (Fic. 3).

The final alignment comprised 763 nucleotides, of which 316 characters

were conserved and 324 were parsimony informative sites. Gaps were treated

as missing data. Tree topologies remained the same in all three phylogenetic

analyses: Maximum Likelihood (ML), Maximum Parsimony, and Nearest

Neighbour Joining).

In the ML phylogenetic tree two different Leucoagaricus sections were

obtained: L. sect. Rubrotincti and L. sect. Piloselli (Fic. 3). The taxa that

clustered together in different sections also have morphological similarities. All

L. lahorensis sequences fall in L. sect. Rubrotincti, along with the sequences of

L. littoralis, L. rubrotinctus, L. sublittoralis sensu Reid, L. vassiljevae, L. wichanskyi,

and other Leucoagaricus species of Vellinga (2001, 2004).

538 ... Qasim & al.

BB Leucoagaricus lahorensis KJ701794 (T38)

IB Leucoagaricus lahorensis KJ701795 (CM36)

BB Leucoagaricus lahorensis KJ701796 (T25)

i Leucoagaricus lahorensis KJ701797 (MCR84)

Leucoagaricus rubrotinctus JX827166 CHINA

Leucoagaricu rubrotinctus FJ481050 CHINA

Leucoagaricus rubrotinctus JN944081 USA

Leucoagaricus sp AY176432 USA

400 Leucoagaricus rubrotinctus JX133167 RUSSIA

Leucoagaricus sublittoralis AY176442 USA

Leucoagaricus vassiljevae JX133169 RUSSIA

100

69 ||Leucoagaricus vassiljevae JX133170 RUSSIA

Leucoagaricus vassiljevae JX896447 RUSSIA

Leucoagaricus vassiljevae JX896445 RUSSIA

Leucoagaricus vassiljevae JX896446 RUSSIA

Leucoagaricus sp JN907015 CHINA

Lepiotaceae sp EF527332 USA Section Rubrotincti

Leucoagaricus crystallifer AF482863 USA

Lepiotaceae sp EF527334 USA

Lepiotaceae sp EF527338 USA

Leucoagaricus littoralis GQ329051 USA

Leucoagaricus littoralis GQ329060 USA

Leucoagaricus wychanskyi AF482874 USA

100

Leucoagaricus purpureolilacinus AF482869 USA

Leucoagaricus purpureolilacinus GQ329053 USA

Leucoagaricus tener GQ329043 USA

Leucoagaricus sp AY176428 USA

Leucoagaricus rubrobrunneus JX133168 RUSSIA

Leucoagaricus rubrobrunneus JX896448 RUSSIA

Leucoagaricus lateritiopurpureus JX133173 RUSSIA

100

Leucoagaricus lateritiopurpureus JX133174 RUSSIA

Leucoagaricus proximus JX133171 RUSSIA

Leucoagaricus proximus JX133172 RUSSIA

gg Leucoagaricus americanus AF295928 USA

Leucoagaricus bresadolae AF295929 USA

a1 Leucoagaricus barssii GQ329062 USA

37 ——-, Leucoagaricus ionidicolor AY 176415 USA

Leucoagaricus variicolor JX880033 ITALY

Leucoagaricus sp Vellinga AY243637 USA

Leucoagaricus sp Vellinga AY243640 USA

Leucoagaricus birnbaumii U85323 USA

Section Piloselli

100

Leucoagaricus fragilissimus U85324 USA

Leucoagaricus melanotrichus GQ329054 USA

Leucoagaricus nympharum JN944087 USA

Leucoagaricus leucothites JQ683123 USA

Leucoagaricus subcretaceus KF410815 India

82 'Leucoagaricus naucinus EU416308 CHINA

Cystolepiota seminuda AY176350 USA _] outgroup

0.05

Fic. 3: Phylogram obtained from ITS region of nrDNA. The phylogenetic relationship of

Leucoagaricus lahorensis was inferred from nrITS sequences using Maximum Likelihood method.

Bootstrap values more than 50% are present above branches. Leucoagaricus lahorensis sequences

are indicated by #.

Leucoagaricus lahorensis sp. nov. (Pakistan) ... 539

Discussion

Basidiomata representing Leucoagaricus lahorensis were collected frequently

from Lahore during the August and September rainy seasons of 2011-13. This

study is based on four collections representing the new taxon: the holotype T25

and MCR84, T38, and CM36. Preliminary morphological analysis suggested

that all four collections represented the same species, which our molecular

analyses confirmed. The ITS-nrDNA based phylogenetic tree also supports

L. lahorensis as separate from all other Leucoagaricus spp. for which ITS data

are available in Genbank.

Leucoagaricus lahorensis is characterized by a reddish brown pileus covering

with white background, radially arranged fibrils becoming sparse towards

margins, close lamellae with entire margins, a white annulus, broadly ellipsoid

basidiospores (occasionally with an amygdaloid apex), narrowly clavate to

sub-cylindrical cheilocystidia without apical crystals, and cylindrical pileal

elements with clavate to narrowly clavate terminal ends.

Our ML phylogenetic tree revealed two Leucoagaricus sections that agree

with those recognized by Singer (1975). All species in L. sect. Rubrotincti are

characterized by a reddish brown pileus covering, ellipsoid spores, tetrasporic

basidia, no colour change upon bruising, and a negative ammonia reaction

(Singer 1948, 1986; Bon 1993; Vellinga 2001).

All L. lahorensis sequences clustered together in L. sect. Rubrotincti with

72% bootstrap support. Leucoagaricus lahorensis forms a sister clade with

Chinese collections tentatively called L. rubrotinctus (66% bootstrap support).

The morphology of these Chinese collections is not known; L. rubrotinctus

(Peck) Singer is an eastern North American species with an orange-red radially

fibrillose pileus (Vellinga 2001). Leucoagaricus lahorensis is distinguished

by its white annulus, broadly ellipsoid spores, and subclavate to cylindrical

cheilocystidia.

Another closely related species is L. vassiljevae E.F. Malysheva et al.

Both species share the approximately same size of pileus (4.5 cm diam for

L. vassiljevae), red brown radially arranged fibrils on a white background, and

broadly ellipsoid dextrinoid basidiospores. However, the large spores (8-13 x

5-6 um), clavate cheilocystidia, and longer stipe (5-13 cm) of L. vassiljevae

distinguish it from L. lahorensis.

Leucoagaricus sublittoralis (Kihner ex Hora) Singer resembles L. lahorensis

in its pinkish brown pileus with white background, 4-spored basidia, and

dextrinoid spores, but its broadly ellipsoid spores and cheilocystidia without

crystals render it a different taxon. Furthermore, L. sublittoralis is found in clay,

loamy soils (Vellinga 2001), while L. lahorensis is found in loamy soils.

540 ... Qasim & al.

Leucoagaricus rufosquamulosus T.K.A. Kumar & Manim. also shows

similarities with L. lahorensis based on brownish red pileus covering, but the

spores with germ pores set it apart from L. lahorensis. Leucoagaricus glabridiscus

(Sundb.) Wuilb. has similarly long spores (10.3 um) (Vellinga 2001) but is

distinguished from L. lahorensis by its 1-2 tiers of lamellulae.

Acknowledgements

We sincerely pay thanks to Dr. Najam-ul-Sahar Afshan and Dr. Amy Y. Rossman for

their valuable comments and for critically reviewing this article. We are also thankful

to Dr. E.C. Vellings for her valuable suggestions to improve the article. This work was

financially supported by University of the Punjab, Lahore, Pakistan.

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fungi. Plant & Soil 170: 63-73. http://dx.doi.org/10.1007/BF02183055

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes — application

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Kirk PM, Canon PF, Minter DW, Stalpers JA. 2008. Dictionary of the Fungi, 10th ed. Wallingford:

CABI.

Kumar TKA, Manimohan P. 2009. The genera Leucoagaricus and Leucocoprinus (Agaricales,

Basidiomycota) in Kerala State, India. Mycotaxon 108: 385-428.

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Liang JE, Yang ZL, Xu J, Ge ZW. 2010. Two new unusual Leucoagaricus species (Agaricaceae)

from tropical China with blue-green staining reactions. Mycologia 102: 1141-1152.

http://dx.doi.org/10.3852/09-021

Malysheva EF, Svetasheva TY, Bulakh EM. 2013. Fungi in the Russian Far East. I. Leucoagaricus

lateritiopurpureus and new species of Leucoagaricus (Agaricaceae) with reddish brown

basidiocarps. Mikologiya i Fitopatologiya 47(3): 169-179.

Mufioz G, Caballero A, Contu M, Vizzini A. 2012. A new Leucoagaricus species of section Piloselli

(Agaricales, Agaricaceae) from Spain. IMA Fungus 2: 117-123.

http://dx.doi.org/10.5598/imafungus.2012.03.02.03

Mufioz G, Caballero A, Contu M, Ercole E, Vizzini A. 2013 [“2014”]. Leucoagaricus croceobasis

(Agaricales, Agaricaceae), a new species of section Piloselli from Spain. Mycological Progress

13(3): 649-655. http://dx.doi.org/10.1007/s11557-013-0947-x

Munsell™. 1975. Soil color charts. Baltimore.

Murrill WA. 1912. The Agaricaceae of the Pacific coast-I. Mycologia 5: 231-262.

http://dx.doi.org/10.2307/3753448

Nawaz R, Khalid AN, Hanif M, Razaq A. 2013. Lepiota vellingana sp. nov. (Basidiomycota,

Agaricales) a new species from Lahore, Pakistan. Mycological Progress 12: 727-732.

http://dx.doi.org/10.1007/s11557-012-0884-0

Singer R. 1948. Diagnoses fungorum novorum Agaricalium. Sydowia 2: 26-42.

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Singer R. 1975. The Agaricales in modern taxonomy, 3rd ed. Vaduz: J. Kramer.

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Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGAS5: Molecular

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Vellinga EC. 2001. Leucoagaricus. 85-108, in: ME Noordeloos et al. (eds). Flora Agaricina

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MY COTAXON

http://dx.doi.org/10.5248/130.543

Volume 130, pp. 543-548 April-June 2015

Aspicilia volcanica, a new saxicolous lichen

from Northeast China

XB +A

GULBOSTAN ISMAYIL)>?, ABDULLA ABBAS , & SHOU-YU GUO3

1 College of Resource and Environment Sciences &

? Arid land Lichen Research Center of Western China, College of Life Science and Technology,

Xinjiang University, Urumqi 830046, P. R. China

3 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, P R. China

* . .

CORRESPONDENCE TO: “guosy@im.ac.cn; *abdula209@sina.com

ABSTRACT — Aspicilia volcanica from Heilongjiang in Northeast China is described as new

to science. The species is characterized by the thin grayish olive thallus, moniliform branched

anastomosing paraphyses, and the presence of stictic and hypostictic acids. It grows on

volcanic rock in temperate region. ITS nrDNA sequence analysis supports the taxonomic

distinctness of this species.

Key worps — Asia, biodiversity, Megasporaceae, Pertusariales, taxonomy

Introduction

The lichen-forming ascomycete genus Aspicilia A. Massal. (Pertusariales,

Megasporaceae) displays a considerable range of morphological variation

(Sohrabi et al. 2011). Within the genus a number of species have radiating

thalli and elongate, often + diverging and + branched marginal areoles, closely

attached to the substratum (Nordin et al. 2011). Aspicilia s. lat (which in the

traditional sense includes Cicinaria, Lobothallia, Sagedia, and Teuvoa) has a

worldwide distribution and covers a diverse assemblage of 200-250 species

(Kirk et al. 2010, Sohrabi et al. 2013). Forty-four species of Aspicilia s. str. (which

includes Teuvoa) have been previously been reported from China (Wei 1991,

Abbas & Wu 1998, Sohrabi et al. 2010, Li et al. 2013), but only two Aspicilia

species (A. subdepressa, A. transbaicalica) were reported from Northeast China

(Wei 1991, Li et al. 2013).

During a study of the lichen genus Aspicilia in China, some interesting

specimens were collected from the Wudalianchi volcanic mountains,

Heilongjiang, Northeast China. Careful morphological and anatomical

544 ... Ismayil, Abbas, & Guo

examination combined with chemical and nrITS sequence analyses has

confirmed one representative, here described Aspicilia volcanica, as new to

science.

Materials & methods

Specimens were collected from the Wudalianchi Scenic Area, Wudalianchi City,

Heilongjiang Province, China, and are preserved in Lichens Research Center in Arid

Zones of Northwest China, Xinjiang University, Urumqi, Xinjiang, China (XJU). The

lichen specimens were examined morphologically using a Leica Zoom 2000 dissecting

microscope and anatomically with an Olympus CH compound microscope. Sections,

mainly cut by hand, were studied in water. Chemical constituents were identified by

thin-layer chromatography using solvent systems A, B, and C (Orange et al. 2010).

Photos of the thallus and anatomical structures were taken with a Nikon Eclipse E200

stereomicroscope with Canon Digital Camera Powershot A640 and Nikon Digital

Camera D50.

DNA EXTRACTION, PCR AMPLIFICATION, & SEQUENCING: DNA was extracted using

DNAsecure Plant DNA Kit (Tiangen, China). PCR amplification followed Martin et al.

(2003) and the manufacturer’s recommendations (Tiangen, China). The whole nrITS

region (ITS1-5.8S-ITS2) was targeted using the primers ITS1-F (Gardes & Bruns 1993)

and ITS4 (White et al. 1990). Thermocycling protocols were 95°C for 3 min linked to

35 cycles at 94°C for 30 s, 54°C for 30 s, and 72°C for 1 min, with a final extension of

72°C for 10 min. PCR products were screened on 1% agarose gels stained with ethidium

bromide and sequenced by the Genewiz Inc. (Beijing).

PHYLOGENETIC ANALYSIS & SEQUENCE COMPARISON: We generated one ITS sequence

(KM609324) and aligned it with 34 representatives (including Lobothallia alphoplaca as

the outgroup) both by ClustalW and Muscle implemented in MEGA 6 (Tamura et al.

2013), then optimized manually. The alignment matrix of 501 nucleotide positions was

submitted to TreeBase (S16691).

A phylogeny was inferred by the Maximum Likelihood method based on the Tamura-

Nei model in MEGA6 and by using Bayesian inference based on GTR model with

rates=Invgamma in MrBayes (Huelsenbeck & Ronquist 2011). For ML analysis, initial

tree(s) for the heuristic search were obtained by applying the Neighbor-Joining method

to a matrix of pairwise distances estimated using the Maximum Composite Likelihood

(MCL) approach. A discrete Gamma distribution was used to model evolutionary rate

differences among sites (5 categories (+G, parameter = 0.4216)).

Results & Discussion

ITS sequence analysis

The ITS sequence analysis supports A. volcanica as a separate species

(Fic. 1) within the A. cinerea-group with close affinities to the common

species, A. cinerea (query cover 100%, 93% identity and 2% gap). Aspicilia

volcanica is also close to A. laevata (query cover 91%, 90% identity, 1% gap) and

A. indissimilis (query cover 92%, 90% identity, 1% gap).

Aspicilia volcanica sp. nov. (China) ... 545

91/1 JF703117 Aspicilia cinerea MARSSJ:Roux 25013

JF703116 Aspicilia cinerea UPS:Nordin 6311

JF703119 Aspicilia cinerea MARSSJ:Roux 25691

JF703114 Aspicilia cinerea UPS:Nordin 5962

EU057899 Aspicilia cinerea UPS:Hermansson 13275

HQ406799 Aspicilia cinerea UPS:Nordin 5542

JF710311 Aspicilia cinerea MARSSJ:Roux 25015

JF703121 Aspicilia cinerea UPS:Owe-Larsson 8914

$3/0.99 JF703120 Aspicilia cinerea H:Sohrabi 3807

9/0.9§ AF332111 Aspicilia cinerea J15

88/IL. JF703118 Aspicilia cinerea MARSSJ:Roux 23869

97/1 24 0b3 AF332112 Aspicilia cinerea J4

1 AF332110 Aspicilia cinerea J12

90/1! JF703115 Aspicilia cinerea UPS:Nordin 6213

79/0.99 KM609324 Aspicilia volcanica sp. nov. from Northeast China

0/0.99 EU057909 Aspicilia indissimilis UPS:Nordin 5943

EU057910 Aspicilia laevata UPS:Tibell 23659

EU057902 Aspicilia sp. Owe-Larsson 9112

99/1 _,»HQ259268 Aspicilia subradians UPS:Nordin 6370

100/1 HQ259267 Aspicilia subradians UPS:Nordin 5984

HQ259264 Aspicilia fluviatilis UPS:Nordin 6188

55/7] 100/1 HQ259265 Aspicilia granulosa UPS:Nordin 6174

HQ259262 Aspicilia epiglypta UPS:Nordin 6105

97/1 & HQ259261 Aspicilia epiglypta UPS:Nordin 6305

99/1 » EU057923 Aspicilia rivulicola UPS:Nordin 5960

EU057922 Aspicilia rivulicola UPS:Nordin 5957

93/1 96/1 p EU057912 Aspicilia mashiginensis UPS:Nordin 5790

HQ259266 Aspicilia mashiginensis UPS:Tibell 23557

HQ259260 Aspicilia dendroplaca UPS:Nordin 6366

53/0.9 99/1 » EU057940 Aspicilia verruculosa UPS:Owe-Larsson 9007

EU057942 Aspicilia verruculosa UPS:Nordin 5942

59/0.98) HQ259269 Aspicilia verruculosa UPS:Roux s.n.

68/- 99/1 » HQ259270 Aspicilia virginea UPS:Nordin 6017a

HQ259271 Aspicilia virginea UPS:Ebbestad SVL1-1

JQ797515 Lobothallia alphoplaca MS130

57/-

739

——|

0.02

FiGuRE 1. Position of the new species in the ITS phylogenetic tree of Aspicilia with Lobothallia

alphoplaca as outgroup. ML bootstrap support / Bayesian posterior probabilities are shown at

nodes; nodes with ML support <50 and PP <0.90 are collapsed. Branch lengths refer to the number

of substitutions per site. Herbarium acronyms (where available) and specimen numbers, and

isolate numbers are attached to the sequences downloaded from GenBank.

Taxonomy

Aspicilia volcanica Ismayil, A. Abbas & S.Y. Guo, sp. nov. FIGURE 2

FUNGAL NAME FEN 570144

Differs from Aspicilia cinerea by its thin grayish olive thallus, the presence of stictic and

hypostictic acids, and the absence of norstictic acid.

Type: China. Heilongjiang: Wudalianchi scenic area, 48°39.321’N 126°09.311’E, alt. 280

m, 15 July 2011, Gulbostan Ismayil & A. Abbas 20111154 (Holotype, HMAS-L; isotype,

XJU; GenBank KM609324).

546 ... Ismayil, Abbas, & Guo

Erymo.oey: The specific epithet volcanica refers to the substrate of the type specimen.

There are 14 volcanic mounts in Wudalianchi, Heilongjiang, Northeast China.

THALLUS grayish olive to olive, areolate to rimose, thin; margin flat, elongated,

branching and diverging, thin, creamy white or whiter than thallus. AREOLEs

irregular to angular and slightly verrucose, flat to + convex, somewhat +

dispersed; separated by cracks and uneven, 0.2-0.5 mm wide and up to 1.2

mm long. CorTEXx paraplectenchymatous, 26-39 um thick, uppermost part

greenish black, without crystals, K-, covered with an epinecral layer, 5-10

uum thick. MEDULLA lax and K-. HypoTHALtus white or whiter than thallus,

smooth at margins. PHoTosionT chlorococcoid, cells + round, 8-18 um in

diam.

APOTHECIA aspicilioid, usually rather common, 1-2 per areole, irregular

and sometimes round or angular, 0.3-0.7 mm in diam. disc black, concave,

epruinose. THALLINE MARGIN flat to elevated in older apothecia, concolorous

with or whiter than thallus, 0.1-0.2 mm thick. EpftHymeNntum blackish or

deep green, without crystals, K+ pale. HyMEeNrum hyaline, 98-143 um tall,

I+ greenish blue to cupper red, K-. HyPOTHECIUM pale, 52-65 um thick, K-,

I+ persistently ink blue. PaRAPHysES moniliform (uppermost part distinctly

moniliform), upper cells globose, branched and anastomosing. Asci Aspicilia-

type, clavate, 8-spored, 57-104 x 16-31 um. Ascosporgs hyaline, simple,

ellipsoid, (13-)16-23(-26) x (10-)13-16 um, with oil drops. Pycnip1a not

found.

SPOT TESTS —K+ greenish yellow, KC-, C-, I-, PD-, UV-.

SECONDARY METABOLITES —Hypostictic acid, connorstictic acid, stictic

acidiG NIE),

EcoLocy —Aspicilia volcanica grows on black rock from volcanic mountain.

Associated species include Rhizocarpon sp., Rhizoplaca sp. At present, it is only

known from the volcanic mountain region in Northeast China at 280-400 m

elevations.

ADDITIONAL SPECIMEN EXAMINED — CHINA. HEILONGJIANG: Wudalianchi,

Wudalianchi Scenic Area, 48°43.213’N 126°27.220’E, alt. 400 m, 13 Jul. 2011, Gulbostan

Ismayil & A. Abbas 20111143 (XJU).

ComMENts — When first collected in the field, A. volcanica was regarded as

representing the common A. cinerea (type species of the genus), which differs by

its gray to almost white thallus with quite large, sometimes confluent apothecia,

a brown to olive-brown epihymenium, slightly shorter ascospores (usually

<20 um long), pycnidia and conidia present, and the presence of norstictic

acid (Owe-Larsson et al. 2007). Aspicilia volcanica is further distinguished

by its smaller somewhat + dispersed areolae, cortex without crystals, smaller

apothecia, and larger ascospores.

Aspicilia volcanica sp. nov. (China) ... 547

FiGuRE 2. Aspicilia volcanica (holotype). A: general habit; B: Apothecia; C: Section of Apothecium;

D: Anastomosing paraphyses; E: Moniliform paraphysis; F: Asci and ascospores. Scale bars: A = 3 mm;

B= 1 mm; C = 200 um; D = 25 um; E = 10 um; F = 20 um.

Previous studies that have used DNA sequence data for species recognition

in lichens required both diagnostic morphological differences and genetic

distance in single-locus nrITS sequence (e.g., Nordin et al. 2011, Han et al.

2013). Our morphological and molecular data for A. volcanica match these

criteria.

548 ... Ismayil, Abbas, & Guo

Acknowledgements

The authors thank Dr. Jan Vondrak (University of South Bohemia, Ceské Budéjovice,

Czech Republic) and Prof. Liu-Fu Han (Hebei Normal University, Shijiazhuang, China)

for presubmission review. S.Y. Guo was awarded grants by the National Natural Science

Foundation of China (nos. 31370067, 30770012) and A. Abbas was awarded grants by

the Natural Science Foundation of China (Nos. 30960003, 31150003). The valuable

assistance given by colleagues is gratefully acknowledged.

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MYCOTAXON

http://dx.doi.org/10.5248/130.549

Volume 130, pp. 549-554 April-June 2015

Redescription of Mycorrhaphium pusillum,

a poorly known hydnoid fungus

KAISA TERVONEN!, VIACHESLAV SPIRIN?, & PANU HALME!?

"Department of Biological and Environmental Science, University of Jyvaskyla;

PO. Box 35, FIN-40014 University of Jyvaskyla, Finland

? Botanical Museum, Finnish Museum of Natural History;

PO. Box 7, FI-00014, University of Helsinki, Finland

° Natural History Museum, University of Jyvaskyla;

PO. Box 35, 40014 University of Jyvaskyla, Finland

* CORRESPONDENCE TO: panu.halme@jyu.fi

ABSTRACT — The hydnoid basidiomycete Mycorrhaphium pusillum was found in Finland.

There are only a few records of the species globally, and the genus Mycorrhaphium has not

been recorded previously in northern Europe. Here we report a detailed description of the

macro- and microscopy of this poorly known species and publish the first photographs of it,

accompanied with detailed microscopic drawings.

Key worps —Mycoleptodonoides, Steccherinaceae, wood pasture

Introduction

During our extensive fungal surveys on Finnish wooded pastures we found

a hydnoid basidiomycete species, Mycorrhaphium pusillum (Maas Geesteranus

1962), from one site in southern boreal vegetation zone. ‘There are no previous

records of the genus Mycorrhaphium in northern Europe. On three occasions

(August 2012; August 2013; August 2014), a single sporocarp was observed

growing in the very same location. Every time we conducted a very thorough

survey of the surroundings, yielding no additional sporocarps in 2012 and 2013

but finding one additional sporocarp some meters from the original location

in 2014. Here we report our findings. First we describe the growth site to

increase the knowledge of the habitat requirements of this rare fungus. Then we

describe the species in more detail than in the original description and discuss

its systematic position.

550 ... Tervonen, Spirin, & Halme

Figure 1. Mycorrhaphium pusillum. Growth site in Luhanka, Finland.

Photograph by Kaisa Tervonen.

Site description

The site is a nutrient rich herb-rich forest dominated by broadleaf tree

species (Fic. 1); Betula spp. predominate, but Alnus incana (L.) Moench, Salix

caprea L., Pinus sylvestris L., Picea abies (L.) H. Karst., Sorbus aucuparia L., and

Rhamnus frangula L. are also present. The site is influenced by calcareous soil.

For example the presence of Russula aurea Pers. as well as the abundance of

Craterellus sinuosus (Fr.) Fr. and the orchid Listera ovata (L.) R. Br. indicates

high soil lime content (Nitare 2000, Mossberg & Stenberg 2005). The site was

slash-and-burn cultivated in the late 19" century and then utilized as a wood

pasture for cattle, probably until the 1970s when it was abandoned; most

recently the area is protected as a habitat of an endangered bird species, white-

backed woodpecker (Dendrocopos leucotos Bechstein). ‘The site hosts especially

rich community of rare fungi, including some wood-inhabiting polypores

(Halme 2008; Markkanen & Halme 2012) as well as ectomycorrhizal and litter

decaying agarics (Tervonen & Halme, unpublished).

Within a ten meter radius of the growth site the most frequent vascular

plant species were Pteridium aquilinum (L.) Kuhn, Calamagrostis arundinacea

(L.) Roth, Hepatica nobilis Schreb., Alnus incana, Rubus saxatilis L., Vaccinium

myrtillus L., Geranium sylvaticum L., and Cirsium helenioides (L.) Hill. The

Mycorrhaphium pusillum redescribed ... 551

most frequent agaric species producing fruit bodies in the same patch were

Lactarius glyciosmus (Fr.) Fr., Cortinarius casimiri (Velen.) Huijsman, a species

of Cortinarius sect. Anomali Bidaud et al., Laccaria laccata (Scop.) Cooke,

Inocybe geophylla (Bull.) P. Kumm., Psathyrella senex (Peck) A.H. Sm., Galerina

triscopa (Fr.) Kihner, Mycena flavoalba (Fr.) Queél., Mycena galericulata (Scop.)

Gray, and Mycena pura (Pers.) P. Kumm.

Methods

Microscopic measurements were made using Olympus and Leica light microscopes

with up to 1600x magnification. Microscopic techniques follow Miettinen et al. (2012b).

Measurements are based on Finnish specimens of M. pusillum. Voucher specimens

are deposited in the Botanical Museum, University of Helsinki, Finland (H) and the

Herbarium, University of Jyvaskyla, Finland (JYV).

Species description

Mycorrhaphium pusillum (Brot.) Maas Geest., Persoonia 2: 398. 1962. Fics 2, 3

BASIDIOCARPS annual, stipitate, leathery, solitary or fusing together (2-3 caps

on common stem). Caps 12-19 mm in diam., evenly rounded and applanate to

FiGuRE 2. Mycorrhaphium pusillum (JYV, Tervonen 3113). Sporocarp:

A. from below, illustrating the spines and stipe; B. from above, illustrating the cap surface.

Photographs by Kaisa Tervonen.

552 ... Tervonen, Spirin, & Halme

asymmetric-flabelliform, pale cream-colored to pale ochraceous, sometimes

with pale brownish zones; margin sharp, even, fertile. Stem 15-21 mm long

and 1-3 mm thick, central or excentric, pale cream. Spines acute, slender,

sometimes fusing together, 0.8-1.2 mm long, 7-8 per mm, tough, pale cream

to pale ochraceous. No distinct smell.

HYPHAL STRUCTURE dimitic in spines, monomitic in context; hyphae

hyaline, faintly cyanophilous.

ConTEXT. Hyphae relatively loosely arranged, with thickened walls,

simple-septate, with clavate terminal portions forming a layer on pileal

surface (“trichoderm”), 3.2—4.8 um in diam., some inflated up to 7 um in diam.

(n = 10/1).

SpineEs. Tramal skeletal hyphae distinctly thick-walled to subsolid (with

capillary lumen), parallel and densely arranged, some with adventive septa,

(3.1-)3.7-4.9(-5.0) um in diam. (n = 20/1); tramal generative hyphae with

distinct walls, clampless, 2.5-3.5 um in diam. Subhymenium distinct (10-15

uum thick), consisting of thin-walled, short-celled, clampless generative hyphae

2-3 um in diam. Gloeocystidia rare, clavate, of subhymenial origin, embedded

in basidial layer, 10.1-11.3 x 4.3-5.4 um. Hyphal pegs infrequent, 10-15 x

15-20 um, consisting of short-celled clampless hyphae 4.1-5.7 um in diam.

Basidia clavate, 1-2(-4)-sterigmatic, clampless, 9-14 x 3-4 um.

Basiprospores thin-walled, thick cylindrical, with more or less flattened

ventral side, tapering to the apiculus, (2.7-)2.9-3.6(-3.8) x (1.8-)1.9-2.2(-2.3) um,

L = 3.03, W = 2.00, Q’ = (1.3-)1.5-1.7(-1.7), Q = 1.52 (n = 60/2), usually with

a prominent oil-drop, inamyloid, acyanophilous.

Hasitat—In herb-rich forest dominated by Betula spp., on mossy, knotty

mull ground. No certain connection with wood was detected.

SPECIMENS EXAMINED: FINLAND. ETEeLA-HAME: Luhanka, Kuruvuori, on ground,

18.VIII.2012, Mustola 1618 & Toivonen (H); 30.VIII.2013, Tervonen 3113 & Sundstrém

(JYV, H). ITALY. PrepMonrt: Turin, Cantalupa, on wood of Fagus sylvatica, 11.III.1984,

Bernicchia 2400 (H).

Discussion

Mycorrhaphium Maas Geest. is now accepted as a small genus comprising

both stipitate and sessile hydnoid fungi, and it was shown to be a member

of the Steccherinaceae (Miettinen et al. 2012a). According to some sources

(www.indexfungorum.org), Mycorrhaphium is regarded as a synonym of

Mycoleptodonoides Nikol., typified with M. vassiljevae Nikol. (Nikolaeva

1952), an East Asian species with a monomitic hyphal structure and allantoid

basidiospores. The author VS studied the holotype of M. vassiljevae (Herb. LE) as

wellas some additional collections from the Russian Far East. Mycoleptodonoides

vassiljevae is a monomitic species having wide, clamped hyphae (in contrast to

Mycorrhaphium pusillum redescribed ... 553

Figure 3. Mycorrhaphium pusillum (H, Mustola 1618).

Skeletal and generative hyphae, basidia, gloeocystidia, and basidiospores.

Drawing by Viacheslav Spirin.

[The characters match the second collection (Tervonen 3113) from the same site. ]

clearly dimitic basidiocarps of the Mycorrhaphium species); in addition, it has

neither the gloeocystidia nor hyphal pegs characteristic of all Mycorrhaphium

species studied by us (including the generic type, M. adustum (Schwein.)

Maas Geest.). Thus we accept the opinion of Maas Geesteranus (1961, 1962)

that Mycoleptodonoides and Mycorrhaphium are two independent, seemingly

not closely related genera. Here we follow the generic concepts proposed by

554 ... Tervonen, Spirin, & Halme

him and consider all dimitic species (including M. pusillum) as members of

Mycorrhaphium. The taxonomic position of Mycoleptodonoides, which is still

unclear, will be clarified in further DNA-based studies.

Mycorrhaphium pusillum is quite small, but easily recognizable and visible

and probably not readily confused with any other species. Still, our record is

the only one in the whole of northern Europe, and M. pusillum has been found

only a few times in central and southern Europe. The site where we recorded

the species is regionally quite unique regarding the stand quality and species

assembly, suggesting that Mycorrhaphium pusillum may be very demanding in

its environmental requirements. Consequently, we believe that this species is

truly rare instead of just being overlooked.

Acknowledgements

We are grateful to Mika Toivonen and Lotta Sundstrém for field assistance and

Tuomo Niemela for assistance in the identification process. We also acknowledge

Tuomo Niemela and Indrek Sell for presubmission review. This study was funded by the

Finnish Ministry of Environment through the Research Programme of Poorly Known

and Threatened Forest Species (PUTTE).

References

Halme P. 2008. Keski-Suomen vanhojen lehtimetsien lahottajasienet. Inventory report,

Metsahallitus. 49 p. (in Finnish).

Maas Geesteranus RA. 1961. A Hydnum from Kashmir. Persoonia, 1: 409-413.

Maas Geesteranus RA. 1962. Hyphal structures in Hydnum. Persoonia, 2: 377-405.

Markkanen A, Halme P. 2012. Polypore communities in broadleaved boreal forests. Silva Fennica,

46: 317-331. http://dx.doi.org/10.14214/sf.43

Miettinen O, Larsson E, Sjékvist E, Larsson KH. 2012a. Comprehensive taxon sampling reveals

unaccounted diversity and morphological plasticity in a group of dimitic polypores (Polyporales,

Basidiomycota). Cladistics, 28: 251-270. http://dx.doi.org/10.1111/j.1096-0031.2011.00380.x

Miettinen O, Spirin V, Niemela T. 2012b. Notes on the genus Aporpium (Auriculariales,

Basidiomycota), with a new species from temperate Europe. Ann. Bot. Fennici 49: 359-368.

http://dx.doi.org/10.5735/085.049.0607

Mossberg B, Stenberg L. 2005. Suuri Pohjolan kasvio. 2nd edition. Tammi. 928 p. (in Finnish).

Nikolaeva TL. 1952. A new genus of hydnoid fungi. Bot. Mater. Otd. Spor. Rast. 8: 117-121.

(in Russian, with Latin subtitle).

Nitare J (ed.). 2000. Signalarter. Indikatorer pa skyddsvard skog. Flora éver kryptogamer.

Skogsstyrelsens Forlag, Jonk6ping. 392 p. (in Swedish).

MYCOTAXON

http://dx.doi.org/10.5248/130.555

Volume 130, pp. 555-561 April-June 2015

Micropsalliota pseudoglobocystis,

a new species from China

Li Wer’, YONG-HE LI’, KEvIN D. HypDE”34, & RuI-LIN ZHAO™*

'Key Laboratory of Forest Disaster Warning & Control in Yunnan Province, Southwest Forestry University,

Kunming 650224, China

?Institute of Agricultural Science, Agricultural Division 5 of Xinjiang Production & Construction Corps,

Bortala, Xinjiang Province, China

*School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

‘Institute of Excellence in Fungal Research, and School of Science, Mae Fah Luang University,

Chiang Rai 57100, Thailand

*The State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academic of Science,

Beijing 100101, China

* CORRESPONDING AUTHOR: Zhaoruilin@gmail.com

ABstRACT — Micropsalliota pseudoglobocystis sp. nov. from China is introduced. The species

is described and illustrated, and its phylogenetic placement is determined using molecular

data. The new species is compared with the most similar taxa in the genus. This is the first

report of the genus Micropsalliota from China.

KEY worps — Agaricaceae, taxonomy, phylogeny

Introduction

Micropsalliota Hohn. (Agaricaceae) was established by Héhnel (1914) and

later emended by Pegler & Rayner (1969) and Heinemann (1976). Micropsalliota

species are often encountered at the sides of forest trails in tropical areas (Zhao et

al. 2010). Micropsalliota is similar to Agaricus L. as both genera have an annulus

on the stipe, free gill attachment, and brown basidiospores. Phylogenetic

analyses of ITS and LSU sequence data have established that Micropsalliota

is monophyletic (Zhao et al. 2010). Micropsalliota can be distinguished from

other similar genera with dark spore prints by its small to medium sized

fruiting bodies, ellipsoid to cymbiform basidiospores with an apical thickening

in the endosporium, often capitate or subcapitate cheilocystidia, and pileipellis

hyphae encrusted with a pigment that turns olive to green in NH,OH

(Zhao et al. 2010).

556 ... Wei & al.

Heinemann (1980, 1983, 1988, 1989), Heinemann & Little Flower (1983),

and Heinemann & Leelavathy (1991) described about 40 species from the

tropics of Africa, America, India, Indonesia, and Malaysia. Zhao et al. (2010)

have since added eleven new taxa to this genus from northern Thailand. There

are presently no reports of Micropsalliota species from China.

A survey of Agaricaceae has been carried out in southwestern China since

2012. Morphological and molecular studies have revealed some undescribed

species, one of which is formally introduced in this paper as Micropsalliota

pseudoglobocystis.

Materials & methods

Morphological examination

Specimens were collected from Tongbiguan National Natural Reserve located at

Yingjiang County, Yunnan Province, China, on 19 July 2013. Photographs were taken

in the field and specimens were wrapped in aluminum foil and kept separately in a

box in order to avoid mixing or crushing. Macromorphological features were described

and macrochemical reactions were made from fresh samples as soon as possible after

returning from the field. Specimens were dried overnight in a food drier, sealed in plastic

bags, and deposited in the herbarium of Southwest Forestry College, Kunming, China

(SWFC) and Herbarium Mycologicum Academiae Sinicae, Beijing, China (HMAS).

Anatomical features (including the pileipellis, partial veil, basidiospores, basidia, and

cystidia) were recorded from dry specimens following the protocols of Largent (1986).

Dimensions were calculated from at least 20 measurements of anatomical features

(spores, basidia and cystidia) and abbreviated as x = mean length by width + SD;

Q = quotient of basidiospore length and width; and Q. = mean of Q-values + SD.

Molecular experiment and phylogenetic analysis

DNA was extracted from the dried specimens using the E.Z.N.A. Forensic DNA

Extraction Kit (D3591-01, Omega Bio-Tek). ITS regions and 5.8S rDNA were amplified

using primers ITS4 and ITS5 according to Zhao et al. (2010) with some modifications.

PCR products were sent to Shuoyang Biotechnology Company for sequencing.

Newly produced sequence and sequences downloaded from GenBank were aligned

using BioEdit v. 7.1.3.0 (http://www.mbio.ncsu.edu/bioedit/bioedit.html) and ClustalX

2.0 in default setting (Thomson et al. 1997). The alignments have been submitted to

TreeBASE (submission ID 16500).

Maximum Parsimony (unweighted) analysis were performed using PAUP*4.0b10

(Swofford 2004). One thousand heuristic searches were conducted with random

sequence addition, with tree bisection-reconnection (TBR) branch swapping and gaps

treated as missing data. Parsimony bootstrap values were obtained from 1000 bootstrap

replicates, with starting trees obtained via stepwise addition, random sequence addition,

TBR branch swapping, and Max-trees set to 1,000,000.

MrModeltest 2.2 was used to determine the appropriate model of nucleotide

substitution for this data for Maximum likelihood analysis and Bayesian inference

(Nylander 2004). Maximum likelihood (ML) analysis was performed in PAUP*4.0b10

Micropsalliota pseudoglobocystis sp. nov. (China) ... 557

HM436637 M. xanthorubescens

HM436636 M. pleurocystidiata

HM436638 M. xanthorubescens

HM436621 M. furfuraceat

HM436624 M. megaspora

HM436623 M. megaspora

HM436627 M. rubrobrunnescens

HM436625 M. rubrobrunnescens

HM436626 M. rubrobrunnescens

HM436629 M. rubrobrunnescensvar. tibiicystis™

HM436628 M. rubrobrunnescens™

60/9:

HM436630 M. brunneospermavar.cortinata

HM436620 M. megarubescens™

HM436618 M. megarubescens

HM436619 M. megarubescens

KM889913 M. pseudoglobocystis™

KM889911M. pseudoglobocystis

KM889912 M. pseudoglobocystis

HM436634 M. globocystis

HM436633 M. globocystis

a HM436635 M. globocystis

_ HM436632M. globoeystis

HM436642 M. bifida

HM436639 M. bifida

HM436640 M. bifida

HM436641 M. bifida

ponies TL HM 436612.M. subarginea

HM436610M. subarginea

73/98|/ HM436611M. subarginea

-— HM436617M. arginophaea

69187 HM436613 M. arginophaea

HM436614 M. arginophaea

L HM436615M@ arginophaea

L HM436616 M. arginophaea

-—— HM436647M. gracilis

HM436646 M. subalba

HM436645 M. pusillissima™

HM436644 M. albosericea

HM436643 M. pseudoarginea

HM436648 M. allantoideaT™

HM436631 M. lateritiavar. vinaceipes™

100/100

75/10

HM436649 Hymenagaricus epipastus

U85307 Agaricus campestris

0.1

FiGuRE 1 Phylogeny of Micropsalliota generated from Maximum Likelihood analysis of ITS

sequences, rooted with Agaricus campestris and Hymenagaricus epipastus. Parsimony bootstrap

support (BS) and Bayesian posterior probability (PP) values >50% are given at the internodes

(BS/PP). “T” indicates a sequence from the type specimen. The new sequences produced from this

research are in bold.

with a GIR+I+G model of nucleotide substitution, starting trees obtained via stepwise

addition, random sequence addition and TBR branch swapping. Bayesian analysis

was performed using Metropolis-coupled MCMC methods in MrBayes 3.1.2 with a

GTR+I+G model and an inv-gamma distribution rate variation across sites. Chains

were run for two parallel searches from random starting trees for 1 million generations

558 ... Wei & al.

and trees were sampled every 100 generations. Those trees sampled prior to searches

reaching a split deviation frequency value reaching 0.01 were discarded as burn-in

(Huelsenbeck & Ronquist 2001; Huelsenbeck et al., 2001; Ronquist & Huelsenbeck

2003).

Results

The ITS dataset included sequences from 43 specimens representing 22 taxa

of Micropsalliota, with Agaricus campestris and Hymenagaricus epipastus as the

outgroup (Zhao et al. 2010). The dataset had an aligned length of 733 characters

in the dataset, of which 122 characters were excluded from all analyses,

389 characters are constant, 87 variable characters are parsimony-uninformative,

and 135 characters are parsimony-informative. Maximum parsimony analysis

recovered one equally parsimonious tree (L = 497 steps, CI = 0.626, RI = 0.806,

RC = 0.504, HI = 0.374). Maximum likelihood analysis produced a single

topology (-InL = 3155.4316; Fic. 1). Bayesian analysis resulted in a single

topology with an average standard deviation of split frequencies = 0.009789.

The ML and Bayesian topologies are nearly identical, differing only in the

position of M. lateritia var. vinaceipes. The ML tree is shown in Fic. 1.

According to the tree, Micropsalliota species form a monophyletic group

with strong support (100BS/100PP values). The three samples cluster

together with strong support (100BS/100PP values), and form a clade sister to

M. megarubescens and M. globocystis.

Taxonomy

Micropsalliota pseudoglobocystis Li Wei & R.L. Zhao, sp. nov. Fic. 2

MycoBank MB 810860

Differs from Micropsalliota globocystis by its smaller basidiospores and its stipe covered

by heavily fibrillose scales.

Type: China, Yunnan Province, Yingjiang County, Tongbiguan National Natural

Reserve, 19 July 2013, collected by Qing-Hua Yu ZRL201332 (Holotype HMAS; isotype,

SWEC; GenBank, KM889913).

ErymMo.oey: the epithet refers to the morphological and phylogenetic similarity to

Micropsalliota globocystis.

PiLEus 25-35 mm diam., broadly and obtusely conical when young, then

expanding to 45-55 mm in diam., convex and subumbonate; margin recurved,

crenate, splitting with age, rarely uplifted; surface dry, covered with dense

fibrillose-scales, recurved, reddish-brown against the whitish background.

FIGURE 2 Micropsalliota pseudoglobocystis (holotype): A, B. Macrocharacters and discoloring

on cutting and bruising; C, D. Cheilocystidia; E. Basidia; F. Basidiospores; G. Annulus hyphae;

H. Pileipellis hyphae.

Micropsalliota pseudoglobocystis sp. nov. (China) ... 559

BW eS tase

sath . a Le

560 ... Wei & al.

CONTEXT 2 mm thick, white. LAMELLAE free, crowded, with 2-4 series of

lamellulae, 2-4 mm broad, at first white, then grayish white, finally dull brown,

edges even entire. STIPE 42-126 x 5-8(-10) mm, cylindrical, sometimes

with basal rhizomorphs, hollow, surface above annulus glabrous to fibrillose,

below annulus floccose, scaly, white. ANNULUS pendent, single, membranous,

superior, edge entire, persistent, white, 4-6 mm broad, upside smooth, lower

side heavily fibrillose. Odor of seaweed. Staining bright yellow then reddish-

brown when bruised or in exposure.

MACROCHEMICAL REACTION: KOH reaction strongly reddish brown, dark

brown in stipe.

BASIDIOSPORES 4.5—6 x 2.5—-3.2 um [x=5.1+0.3 x 2.8+0.2,Q=1.6-2.1,Q =1.8

+ 0.13, n = 20], ellipsoid, amygdaliform, with apical thickening (endosporium),

no germ pore, brown. Basip1A 15-20 x 5-7 um, clavate, hyaline, 4-spored.

CHEILOCYSTIDIA 40-50 x 10-13 um, irregularly cylindrical to subclavate with

a subcapitate to capitates apex, and often present a long, narrow and flexuous

neck, smooth, hyaline. PLEUROCysTIDIA absent. PILEIPELLIS a cutis composed

of hyphae 7-18 um diam., constricted at the septa, incrusted, contains brown

membranous pigments. ANNULUS composed of hyphae of 5-10 um diam.

smooth, hyaline and branched.

Hasir: gregarious on the bank of a forest trail.

ADDITIONAL MATERIAL EXAMINED: CHINA, YUNNAN PROVINCE, /Yingjiang

County, Tongbiguan National Natural Reserve, 19 July 2013, collected by Qing-Hua

Yu ZRL2013323 (SWFC; GenBank KM889912); ZRL2013335 (SWFC; GenBank

KM889911).

Discussion

Other species of Micropsalliota with reddish-brown or purple brown scales

on the pileus are M. arginophaea Heinem., M. furfuracea R.L. Zhao et al.,

M. megaspora R.L. Zhao et al., M. pleurocystidiata Heinem. & Little Flower,

M. xanthorubescens Heinem., and M. globocystis Heinem. (Heinemann 1976,

1980; Zhao et al. 2010). All of these species differ from M. pseudoglobocystis

morphologically, as well as phylogenetically: M. arginophaea does not discolor

on bruising or cutting and has brown pileus scales; M. furfuracea has larger

pileus scales; M. megaspora has larger basidiospores (6-7 x 3.5-4 um);

M. pleurocystidiata has pleurocystidia; M. xanthorubescens has medium-sized

basidiomata (pileus 40-70 mm diam.); and M. globocystis has distinctly larger

basidiospores (6-8 x 3.5-4.2 um) and its stipe has a tomentose surface. Another

phylogenetically closely related species is M. megarubescens R.L. Zhao et al.,

which differs by being slightly fibrillose on the disc of the pileus and glabrous

elsewhere (Zhao et al. 2010).

Micropsalliota pseudoglobocystis sp. nov. (China) ... 561

Acknowledgements

The authors thank Kanad Das (Botanical Survey of India, Howrah, India) and

Zai-Wei Ge (Kunming Institute of Botany, Chinese Academy of Sciences, China) for

pre-submission review. The National Natural Science Foundation of China (Project ID:

31000013, 31360014 and 31470152 to Rui-Lin Zhao) are acknowledged for financial

support this study.

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Zhao R, Desjardin DE, Soytong K, Perry BA, Hyde KD. 2010. A monograph of Micropsalliota in

Northern Thailand based on morphological and molecular data. Fungal Divers. 45: 33-79.

http://dx.doi.org/10.1007/s13225-010-0050-4

MY COTAXON

http://dx.doi.org/10.5248/130.563

Volume 130, pp. 563-567 April-June 2015

Two new Rosellinia species from Southwest China

QiRuI LI’, JICHUAN KANG”, & KEVIN D. HYDE?

'The Engineering and Research Center for Southwest Bio-Pharmaceutical Resources of National

Education Ministry of China, Guizhou University, Guiyang 550025, PR China

*Centre of Excellence in Fungal Research, and School of Science, Mae Fah Luang University,

Chiang Rai 57100, Thailand

*CORRESPONDENCE TO: bcec.jckang@gzu.edu.cn

ABSTRACT— Two new species of Rosellinia from China are illustrated and described.

Rosellinia sigmoidea differs from other species mainly in having a white entostroma and

broadly rounded ascospores with sigmoid germ slits extending over half of the spore length.

Rosellinia camphorae is unique because of its large ascus apical apparatus and ascospores with

slimy sheaths. The herbarium and living culture are deposited in the Collection of Guizhou

University (GZUC).

Key worps— Ascomycetes, Pezizomycotina, taxonomy, Xylariales, Xylariaceae

Introduction

Rosellinia De Not. (Xylariaceae), proposed in 1844 with R. aquila (Fr.)

De Not. as its type species, is an important cosmopolitan genus. Stromata

of Rosellinia are usually uniperitheciate and embedded in a persistent or

fugacious subiculum. The ascus apical apparatus is well developed, especially

in species with large ascospores. The asexual morph of Rosellinia species can be

assigned to Dematophora R. Hartig, Geniculosporium Chesters & Greenh., or

Nodulisporium Preuss (Greenhalgh & Chesters 1968, Rogers & Malmgren 1977,

Petrini 1992, Stadler et al. 2013, Maharachchikumbura et al. 2015). Conidia

are observed in the subiculum, on immature perithecia or in culture (Petrini

& Petrini 2012). According to Petrini (2003) and Petrini & Petrini (2005),

ascospore morphology is the most stable morphological character to delineate

species. Currently, 142 species are accepted by Petrini (2013). Rosellinia

species have been found on wood, dicotyledonous plants, and (occasionally)

monocotyledonous hosts (Martin 1967; Pande & Rao 1995; Petrini 1992, 2003;

Petrini et al. 1989; Roger 1953, Rogers 1979). Many Rosellinia species cause

plant disease (Whalley 1996).

564 ... Li, Kang, & Hyde

Materials & methods

The fresh specimen was collected from Guizhou Province, China. Asci and ascospores

were examined by light microscopy (BX41, Olympus). Material was mounted in water

and Melzer’s iodine reagent for examination. At least 20 propagules were measured to

establish length and width ranges. Bubbles and spots have been removed from images.

Taxonomy

Rosellinia sigmoidea QR. Li, J.C. Kang, K.D. Hyde, sp. nov. PLATE 1

MycoBank MB 810825

Differs from all other Rosellinia species by its white entostroma and its broadly rounded

ascospores with a sigmoid germ slit extending over half the spore length.

Type: China, Guizhou Prov., Guiyang, on dead wood, March 2014, Qirui Li (Holotype,

GZUHO0105).

Erymo ocy: The epithet refers to the sigmoid germ slit of ascospores.

PLaTE 1. Rosellinia sigmoidea (holotype, GZUH0105). A, B. Stromata on the host. C. Section of

stroma. D-G. Asci. H, I. Urn-shaped, J+, apical apparatus. J-O. Ascospores. Scale bars: B = 500 um;

C = 200 um; D-I = 10 um; J-O = 5 um.

Rosellinia spp. nov. (China) ... 565

SAPROBIC On woody material. Sexual morph: SuUBICULUM evanescent, brown to

black, gradually disappearing, absent in mature material. Srromara black, 0.4-

0.8 mm diam., subglobose to globose, with a central ostiole, solitary or densely

crowded in small groups. Ecrostroma <90 um thick, black. ENrosrromaA

white. AscomaTa 350-600 um high, 310-600 um wide, easy detached.

PARAPHYSES persistent at maturity, hypha-like, tapering slightly towards

the rounded apex. Asci 116.5-224.5 x 10-15 um (mean = 157.1 x 12.1 um,

n = 20), 8-spored, unitunicate, cylindrical, pedicellate, apically rounded, with

an amyloid apical apparatus. APICAL APPARATUS barrel-shaped, 3-5 um high

(mean = 4.3 um, n = 20), upper width 3.5-5 um (mean = 3.9 um, n = 20),

lower width 2.5-4 um (mean = 3 um, n = 20). AscosporEs 12.5-15 x 6-7.5 um

(mean = 13.6 x 6.9 um, n = 30), uniseriate, ellipsoidal, ends rounded, dark

brown to black at maturity, unicellular, smnooth-walled, with sigmoid germ slit

about the half spore length, lacking sheath. Asexual morph: undetermined.

ComMENTS—Rosellinia lakshadweepensis A. Pande & V.G. Rao differs from

R. sigmoidea in its ascospores being surrounded by a slimy sheath; and

R. cibodasae L.E. Petrini differs by its larger ascospores (21.9 x 6.8 um) (Petrini

& Petrini 2005, Petrini 2013).

Rosellinia camphorae Q.R. Li, J.C. Kang, K.D. Hyde, sp. nov. PLATE 2

MycoBAank MB 810826

Differs from Rosellinia procera by its larger apical apparatus and its ascospores lacking

caps at the ends and having a germ slit their entire length.

Type: China, Guizhou Prov., Guiyang, on deadwood of Camphora sp., March 2014,

Qirui Li (Holotype, GZUH0113).

Erymo.oey: The epithet refers to the host genus, Camphora.

SAPROBIC On woody material. Sexual morph: SUBICULUM evanescent, brown

to black, gradually disappearing, absent in mature material. Srromata black,

0.8-1.2 mm high, 1-1.5 mm wide, subglobose to globose, with a central ostiole,

solitary or densely crowded in small groups. ECTOsTROMA up to 60 um thick,

black. ENTosTROMA disappearing at maturity. AscomaTA 500-1000 um diam.,

500-1000um high, black. PARAPHYSES persistent at maturity, hypha-like, tapering

slightly toward the rounded apex. Asci 257-288 x 35-50 um (265 x 41 um,

n = 20), 8-spored, unitunicate, clavate, short pedicellate, apically rounded, with

a large amyloid apical apparatus. APICAL APPARATUS barrel-shaped, 20-28.5 um

high, upper width 9-11 um (9.5 um, n = 20), lower width 14-16 um (15.3 um,

n = 20). Ascospores 89-105 x 14.5-19 um (99.5 x 17.2 um, n = 20), bi-seriate,

fusiform, ends rounded, dark brown to black at maturity, unicellular, smooth-

walled, with a germ slit running the entire length of the spore, possessing a

thin, slimy sheath. Asexual morph: undetermined.

566 ... Li, Kang, & Hyde

mo, M

PLATE 2. Rosellinia camphorae (holotype, GZUH0113). A, B. Stromata on the host. C. Section

of stroma. D. Paraphyses. E-H. Asci. I, J. Urn-shaped, J+, apical apparatus. K-M. Ascospores.

Scale bars: B, C = 500 um; D = 50 um; E-M = 10 um.

ComMMENTS—Rosellinia camphorae is similar to R. procera Syd. & P. Syd.,

R. megalosperma Syd. & P. Syd. R. formosana Y.M. Ju & J.D. Rogers,

R. markhamiae Sivan., and R. saccasii L.E. Petrini in ascospore size. However,

R. procera differs by its smaller ascal apical apparatus (13-15 um high,

8.5-10 um broad) and its ascospores having caps at the ends, but lacking a

germ slit (San Martin & Rogers 1995, Petrini & Petrini 2005, Petrini 2013);

R. megalosperma differs by its ascospores having caps at the ends (Petrini &

Petrini 2005); and R. formosana, R. markhamiae and R. saccasii differ by their

smaller apical apparatus (Petrini 2013).

Acknowledgments

The authors are grateful for pre-submission comments and suggestions provided

by Yucheng Dai and Xiuguo Zhang. This work was funded by the grants from the

Rosellinia spp. nov. (China) ... 567

National Natural Science Foundation of China (NSFC, No. 30870009), the international

collaboration plan of Guizhou Province (No. G [2012]7006) and the innovation team

construction for science and technology of Guizhou province (No. [2012]4007) from

the Science and Technology Department of Guizhou province, China.

Literature cited

Greenhalgh GN, Chesters CGG. 1968. Conidiophore morphology in some British members of the

Xylariaceae. Trans. Br. Mycol. Soc. 51: 57-82. http://dx.doi.org/10.1016/s0007-1536(68)80122-6

Maharachchikumbura SS, Hyde KD, Jones EBG, McKenzie EHC, Huang SK, Abdel-Wahab MA,

Daranagama DA, Dayarathne M, D’souza MJ, Goonasekara ID, Hongsanan S, Jayawardena

RS, Kirk PM, Konta S, Liu JK, Liu ZY, Norphanphoun C, Pang KL, Perera RH, Senanayake

IC, Shang Q, Shenoy BD, Xiao YP, Bahkali AH, Kang JC, Somrothipol S, Suetrong S,

Wen TC, Xu JC. 2015. Towards a natural classification and backbone tree for Sordariomycetes.

Fungal Diversity 72: 199-301.

Martin P. 1967. Studies in the Xylariaceae: Il. Rosellinia and the Primo-Cinerea section of

Hypoxylon. J. South Afr. Bot. 33: 315-328.

Pande A, Rao G. 1995. The genus Rosellinia (Sphaeriales) form Peninsular India. Czech Mycology

48: 177-182.

Petrini LE. 1992. Rosellinia species of the temperate zones. Sydowia 44: 169-281.

Petrini LE. 2003. Rosellinia and related genera in New Zealand. New Zealand J. Bot. 41: 71-138.

http://dx.doi.org/10.1080/0028825X.2003.9512833

Petrini LE. 2013. Rosellinia: a world monograph. Gebruder Borntraeger Verlagsbuchhandlung,

Science Publishers.

Petrini LE, Petrini O. 2005. Morphological studies in Rosellinia (Xylariaceae): the first step towards

a polyphasic taxonomy. Mycol. Res. 109(5): 569-580.

http://dx.doi.org/10.1017/S0953756205002510

Petrini LE, Petrini O. 2012. Rosellinia species (Xylariaceae) from South and Central America.

An annotated list. Kurtziana 37(1): 127-139.

Petrini LE, Petrini O, Francis SM. 1989. On Rosellinia mammaeformis and other related species.

Sydowia 41: 257-276.

Roger L. 1953. Genre Rosellinia De Not. Phytopathologie des Pays Chauds 2: 1286-1297.

Rogers JD. 1979. The Xylariaceae: systematic, biological and evolutionary aspects. Mycologia 71:

1-42. http://dx.doi.org/10.2307/3759218

Rogers JD, Malmgren MM. 1977. Notes on Rosellinia buxi and conidial Xylaria. Can. J. Bot. 55:

1051-1055. http://dx.doi.org/10.1139/b77-123

San Martin GE, Rogers JD. 1995. Rosellinia and Thamnomyces in Mexico. Mycotaxon 53: 115-127.

Stadler M, Kuhnert E, PerSoh D, Fournier J. 2013. The Xylariaceae as model example for a unified

nomenclature following the “One fungus-one name” (1F1N) concept. Mycology 4(1): 5-21

Whalley AJS. 1996. The xylariaceous way of life. Mycol. Res. 100: 897-922.

http://dx.doi.org/10.1016/S0953-7562(96)80042-6

MY COTAXON

http://dx.doi.org/10.5248/130.569

Volume 130, pp. 569-575 April-June 2015

Some new records of Uredinales

from Khyber Pakhtunkhwa, Pakistan

M. Fiaz', H. AHMAD’, N.S. AFSHAN?*, & A.N. KHALID‘4

"Department of Botany & *Department of Genetics, Hazara University, Mansehra, Pakistan

3*Centre for Undergraduate Studies & *Department of Botany,

University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

AsBstTRAcT — Uromyces ferulae and the telial stage of Cerotelium fici were collected from

Khyber Pakhtunkhwa and are new records for Pakistan. Puccinia exhausta on Clematis grata

and P. ustalis on Ranunculus hirtellus are reported as new host records for these rusts in

Pakistan.

Key worps —Oghi forest, Pucciniales, rust fungi

Introduction

During the exploration of Uredinales of Khyber Pakhtunkhwa, Pakistan,

rust-infected plants were collected from different localities and examined

macro-microscopically. Among the specimens, Heracleum cachemiricum was

found infected with Uromyces ferulae, which is a new record for Pakistan. The

telial stage of Cerotelium fici on Ficus carica and F. palmata is an addition to

the already reported stages of this rust from Pakistan. Puccinia exhausta on

Clematis grata and P. ustalis on Ranunculus hirtellus are new host records

for these rusts in Pakistan. This work brings the total of rust taxa recorded

from Khyber Pakhtunkhwa to 174. Previously about 173 species of rust fungi

have been reported from this area (Afshan & Khalid 2008, 2009; Afshan et al.

2008a,b,c,d, 2010; Ishaq et al. 2013).

Materials & methods

Freehand sections & scrape mounts of infected plant materials were made in lactic

acid. The plants were photographed and infected portions were observed under a

stereomicroscope. Twenty spores representing each spore state were examined under

a Nikon YS 100 microscope, and paraphyses and spore were measured using a Zeiss

570 ... Fiaz & al.

eyepiece screw micrometer. Sections, paraphyses and spores were microphotographed

by Digiporo-Labomed. Spores and paraphyses were drawn with the aid of a camera

lucida (Ernst Leitz Wetzlar Germany). Specimens are conserved in the Herbarium,

Hazara University, Dhodial, Pakistan (HUP).

New records

Uromyces ferulae Juel, Bull. Soc. mycol. France 17: 259. 1901. PLaTE 1

SPERMOGONIA and AECIA were not found. UREDINIA and TELIA are

mixed in the same sori, amphigenous and petiicolous, rounded, in dense

clusters, especially on the leaflets, dark brown 0.09-0.26 x 0.25-0.41 mm.

UREDINIOSPORES globoid or subgloboid, 22.4-24.1 x 26.9-27.9 um (mean

23.4-27.6 um), hyaline to golden brown, echinulate; germ pores 2-4, equatorial;

pedicel hyaline, deciduous. TeELIosporeEs globose to subglobose or ovoid to

ellipsoid, 17.7-23.7 x 29.3-37.8 um (mean 22.2-33.4 um), apex rounded; wall

1.4-2.6 um, light brown to chestnut brown, smooth; apex 2.8-5.2 um thick,

germ pore mostly at the apex, pedicel hyaline, deciduous.

bd,

bre Ne 5

so yaeet

PLaTE 1: Uromyces ferulae (HUP MFR-281):

A. Teliospores. B. Urediniospores. Scale bars = 10 um.

Rusts and hosts new for Pakistan ... 571

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWaA, District Mansehra, Oghi,

Khabbal Paien, at 2046 m a.s.l., on Heracleum cachemiricum C.B. Clarke (Umbelliferae),

stages II + III, May 2010, M. Fiaz #FR-208 (HUP MFR-281).

ComMENTts: Uromyces ferulae has been reported on different Ferula species

from North Africa, southern Europe, Australia, and Asia (Iran) (Saccardo

& Saccardo 1905: 248; Mercé 1975; Denchev 1995). In Pakistan, no species

of Uromyces has previously been recorded on Umbelliferae. This is the first

report from Pakistan, and Heracleum cachemiricum represents a new host for

Uromyces ferulae.

Cerotelium fici (Castagne) Arthur, Bull. Torrey Bot. Club 44: 509. 1917. PLATE 2

SPERMOGONIA and AECIA are unknown. UrREDINIA hypophyllous, scattered

over the whole surface, minute, sometimes in large irregular groups, slightly

PLATE 2: Cerotelium fici (HUF MFR-252): C. Subepidermal telium. D. Incurved paraphyses.

E. Urediniospores. Scale bars: C = 9 cm; D = 8.5 um; E = 6 um.

572 ... Fiaz & al.

erumpent, pulverulent, reddish brown, surrounded by peripheral, incurved

and basally united paraphyses. UREDINIOSPORES subglobose or broadly ellipsoid to

obovoid or ovate, densely echinulate, yellowish brown, 16-21.7 x 17-31.2 um,

wall 1-1.6 um thick, germ pores scattered, obscure. TELIA hypophyllous,

subepidermal, scattered, minute, 190-270 um in diam. TELIOSPORES

one-celled, cylindrical, broadly ellipsoid or oblong, smooth, 8.7-11.4 x

14.6-20.8 um, catenulate, in two or three superimposed layers, walls uniformly

thin, colorless.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, District Mansehra,

Bherkund, at 849 m a.s.l., on Ficus carica L. (Moraceae), stages II + III, Nov. 2010, M.

Fiaz #FR-093 (HUP MFR-252); District Mansehra, Gulebagh, at 920 m a.s.l., on Ficus

palmata Forssk., stage IH, Oct. 2009, M. Fiaz #FR-089 (HUP MFR-253).

Comments: The telial stage of Cerotelium fici is a new record for Pakistan. The

uredinial stage of Cerotelium fici has been reported on Ficus carica, FE palmata,

and F. religiosa L. from Lahore, Change Manga, Sangla Hill, Tandojam, Malir

(Karachi), and Rawalpindi (Ahmad 1956a,b, Hasnain et al. 1959, Khan &

Kamal 1968, Ghaffar & Kafi 1968, Kaneko 1993).

Puccinia exhausta Dietel, Bot. Jb. 28: 283. 1900. PLATE 3

SPERMOGONIA are usually epiphyllous, subcutaneous, chestnut-brown,

spherical or ellipsoid, 74-117 x 79-126 um. AEcra and UREDINIA not found.

TELIA amphigenous, small, rounded, scattered or in circular groups, dark

brown to blackish, erumpent or covered by epidermis. TELIOsPORES mostly

ellipsoid, sometimes 1-celled and elongated, conical or rounded at the apex,

base mostly roundish, apical wall may be thickened up to 3 um, sometimes

2-celled, not thickened, slightly constricted or non-constricted at the septum,

smooth, hyaline to yellowish and chestnut brown, 17-24 x 26-47 um (mean

= 21 x 39 um), wall 1.4-2.5 um, germ pore apical in upper cell, adjacent to

the pedicel or basal in lower cell, distinct colorless papilla over the germ pore,

sometimes up to 4 um; pedicels hyaline, deciduous, rarely up to 80 um long.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, District Mansehra,

Dhodial, at 978 m a.s.l., on Clematis grata Wall. (Ranunculaceae), stages 0 + III, Aug.

2010, M. Fiaz #FR-003. (HUP MFR-266).

COMMENTS: Puccinia exhausta is cosmopolitan and reported on different

Clematis species (Hiratsuka & Chen 1991, Hiratsuka et al. 1992, Kobayashi

2007). From Pakistan, it has previously been reported on Clematis montana

Buch.-Ham. ex DC. from the Kawai and Malakandi forests (Kaghan valley) by

Ono & Kakishima (1992) and Ono (1992). Clematis grata represents a new host

record for P. exhausta in Pakistan.

Rusts and hosts new for Pakistan ... 573

PLATE 3: Puccinia exhausta (HUP MFR-266): Teliospores. Scale bar = 10 um.

Puccinia ustalis Berk. Hooker’s J. Bot. Kew Gard. Misc. 6: 207. 1854. PLATE 4

SPERMOGONIA, AECIA and UREDINIA are unknown. TELIA usually

hypophyllous as dark brown to blackish spots, scattered or aggregated in large

groups, compact. TELIOSPORES mostly 2-celled, occasionally 1- or 3-celled

teliospores co-exist, ellipsoid, oblong to cylindrical or irregular, conical or

rounded above and narrowed below, constricted at the septum, yellowish brown,

pale brown basally; 2-celled spores 10-16 x 33-63 um, smooth, 1-1.5 um

thick at side, apically 2-8.6 um thick; germ pore one per cell, apical or sub

apical in distal cell, in basal cell near the septum or obscure; 1-celled spores

8-11 x 17-33 um, at side 1-1.5 um, apices 2—-5.4 um thick; 3-celled spores 10-13

x 48-52 um, 1-1.5 um at sides and 2-6 um at the apices. Pedicel pale yellow to

hyaline, deciduous, 7-10 um long.

57A ... Fiaz & al.

PiatE 4: Puccinia ustalis (HUF MFR-272): Teliospores. Scale bar = 12 um.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, District Mansehra,

Balakot, Nadibanglaw, at 2515 m a.s.l., on Ranunculus hirtellus Royle (Ranunculaceae),

stage III, Jul. 2011, M. Fiaz #FR-005. (HUP MFR-272).

CoMMENTS: Puccinia ustalis is cosmopolitan and reported on different species

of Anemone, Clematis, Aquilegia, Pulsatilla, Ranunculus, Thalictrum, and

Trautvetteria (Hiratsuka et al. 1992). From Pakistan, it has been reported on

Anemone obtusiloba D. Don from Changla Gali and Nathia Gali (Ahmad

1956a,b) and on Ranunculus diffusus DC. from Dunga Gali (Okane et al. 1992)

and the Kaghan Valley (Ono 1992). Ranunculus hirtellus represents a new host

record for Puccinia ustalis in Pakistan.

Acknowledgements

We sincerely thank Dr. Abdul Rehman Niazi (Department of Botany, University

of the Punjab, Lahore, Pakistan) and Dr. Omar Paino Perdomo (Dominican Society

of Mycology, Santo Domingo, Dominican Republic) for their valuable suggestions to

improve the manuscript and acting as presubmission reviewers.

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20(1): 82-87.

Afshan NS, Khalid AN. 2009. New records of Puccinia and Pucciniastrum from Pakistan.

Mycotaxon 108: 137-146. http://dx.doi.org/10.5248/108.137

Rusts and hosts new for Pakistan ... 575

Afshan NS, Khalid AN, Niazi AR. 2008a. New records and distribution of rust fungi from Pakistan.

Mycotaxon 105: 257-267.

Afshan NS, Khalid AN, Niazi AR. 2008b. New records of graminicolous rust fungi from Pakistan.

Pak. J. Bot. 40(3): 1279-1283.

Afshan NS, Khalid AN, Javed H. 2008c. Further addition to the rust flora of Pakistan. Pak. J. Bot.

40(3): 1285-1289.

Afshan NS, Berndt R, Khalid AN, Niazi AR. 2008d. New graminicolous rust fungi from Pakistan.

Mycotaxon 104: 123-130.

Afshan NS, Niazi AR, Khalid AN. 2010. Three new species of rust fungi from Pakistan. Mycol.

Progress 9: 485-490. http://dx.doi.org/10.1007/s11557-010-0655-8

Ahmad S. 1956a. Uredinales of Pakistan. Biologia 2: 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biol. Soc. Pak., Lahore.

Denchev CM. 1995. Bulgarian Uredinales. Mycotaxon 55: 405-465.

Ghaffar A, Kafi A. 1968. Fungi of Karachi. Pak. J. Bot. 4: 195-208.

Hasnain SZ, Khan A, Zaidi AJ. 1959. Rusts and smuts of Karachi. Bot. Dept. Karachi Univ. Mont.

Hiratsuka N, Chen ZC. 1991. A list of Uredinales collected from Taiwan. Trans. Mycol. Soc. Japan

3213-22.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba Shuppankai, Tsukuba.

Ishaq A, Afshan NS, Khalid AN. 2013. New records of Puccinia on Poaceae from Khyber

Pakhtunkhwa, Pakistan. Mycotaxon 126: 177-182. http://dx.doi.org/10.5248/126.177

Kaneko S. 1993. Parasitic fungi on woody plants from Pakistan. Cryptogamic Flora of Pakistan,

Vol. 2 (Eds. T. Nakaike and S. Malik), Nat. Sci. Mus., Tokyo, Japan. pp. 149-168.

Khan SA, Kamal M. 1968. The fungi of South West Pakistan. Part 1. Pak. J. Sci. & Ind. Res. 11: 61-80.

Kobayashi T. 2007. Index of fungi inhabiting woody plants in Japan. Host, Distribution and

Literature. Zenkoku-Noson-Kyoiku Kyokai Publishing Co., Ltd.

Mercé J. 1975. Royas de umbeliferas de Las Sierras Andaluzas. Lagascalia 5: 27-34.

Okane I, Kakishima M, Ono Y. 1992. Uredinales collected in Murree Hills, Pakistan. Cryptogamic

flora of Pakistan 1: 185-196.

Ono Y. 1992. Uredinales collected in the Kaghan Valley, Pakistan. Cryptogamic flora of Pakistan

1: 217-240.

Ono Y, Kakishima M. 1992. Uredinales collected in the Swat Valley, Pakistan. Cryptogamic flora of

Pakistan 1: 197-216.

Saccardo PA, Saccardo D. 1905. Supplementum universale pars VI. Hymenomycetae-

Laboulbeniomycetae. Sylloge Fungorum 17. 991 p.

MY COTAXON

http://dx.doi.org/10.5248/130.577

Volume 130, pp. 577-591 April-June 2015

The genus Allocetraria (Parmeliaceae) in China

Ru1i-FANG WANG??, XIN-LI WEI’, & JIANG-CHUN WEI?

' State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing, 100101, China

? College of Life Sciences, Shandong Agricultural University, Taian, 271000, China

* CORRESPONDENCE TO: weixl@im.ac.cn, weijc2004@126.com

ABSTRACT—Ten species of Allocetraria are reported from China, including the new species

Allocetraria corrugata, which is characterized by strongly rugose upper and lower lobe

surfaces. A phylogenetic analysis based on nrDNA ITS sequences supports the independence

of the new taxon. Diagnostic characters and distribution of the Allocetraria species occurring

in China are given, and a key to the identification of the species is provided.

KEY worps— cetrarioid lichens, chemistry, comprehensive analysis, morphology, taxonomy

Introduction

The lichenized genus Allocetraria Kurok. & M.J. Lai, which was described

in 1991, comprised two species from other genera (the type, A. stracheyi,

and A. ambigua) and one new species, A. isidiigera. The genus was originally

characterized by dichotomously or subdichotomously branched lobes and

foliose to suberect or erect thallus with sparse rhizines, angular to sublinear

pseudocyphellae, a palisade plectenchymatous upper cortex, and the presence

of usnic acid in the cortex (Kurokawa & Lai 1991). Randlane & Saag (1992) later

transferred three additional taxa—A. cucullata, A. nivalis, and A. potaninii—

based on morphological, anatomical, and chemical data. Subsequently,

Karnefelt et al. (1994) transferred A. cucullata and A. nivalis into a new genus

Flavocetraria, distinguished from Allocetraria based on ascus structure and

ascospore morphology.

Thell et al. (1995) described two additional new Allocetraria species and

transferred three species from other genera—A. flavonigrescens, A. sinensis,

A. denticulata, A. globulans, and A. oakesiana—and synonymized A. potaninii

with A. stracheyi. At that time the genus was diagnosed as having a palisade

plectenchymatous cortex, asci with a very broad axial body, globose or

578 ... Wang, Wei, & Wei

subglobose ascospores, and filiform conidia (Thell et al. 1995). Later, Karnefelt &

Thell (1996) transferred two additional species to Allocetraria—A. endochrysea

and A. madreporiformis.

Lai et al. (2007) established a new genus Usnocetraria and transferred

numerous Allocetraria species into the new genus (all but two as invalidly

published combinations). However, Thell et al. (2009) demonstrated that none

of these species was closely related to the type species Usnocetraria oakesiana

[= Allocetraria oakesiana]. Recently, Wang et al. (2014) described a new species

Allocetraria capitata characterized by having capitate soralia on the top of lobes.

During our taxonomic study of Allocetraria, we have identified an additional

new species, which brings to ten the number of species accepted in the genus

Allocetraria.

In its current circumscription Allocetraria is a well-supported monophyletic

group within the cetrarioid clade (Saag et al. 2002, Thell et al. 2009, Nelsen et

al. 2011), and China is the distribution center of the genus, with all ten species

known to occur in China. An overview and a key to all ten Allocetraria species

are provided below.

Materials & methods

Phenotypic study

A total of over 1000 specimens of Allocetraria from mainland China were examined.

A dissecting microscope (Zeiss Stemi SV11) and compound microscope (Zeiss

Axioskop 2 plus) were used to study the morphology and anatomy. All specimens are

conserved in Herbarium Mycologium, Institute of Microbiology, Chinese Academy of

Sciences, Beijing, China (HMAS).

Color test reagents (10% aqueous KOH, saturated aqueous Ca(OCl),, and

concentrated alcoholic p-phenylenediamine) and thin-layer chromatography (TLC,

solvent system C) were used to detect lichen substances (Culberson & Kristinsson 1970,

Culberson 1972).

DNA extraction & PCR amplification

Twenty-seven freshly collected lichen specimens were chosen for DNA extraction

(TABLE 1). The extraction procedure followed a modified CTAB method (Rogers &

Bendich 1988). PCR amplifications were performed using a Biometra T-Gradient

thermal cycler. The primer pairs ITS1 (White et al. 1990) and LR1 (Vilgalys & Hester

1990) were used to amplify the nrDNA ITS region. Reactions were carried out in

25 ul reaction volumes containing 0.5 uL total DNA, 1 uL each primer (10uM), 0.5

uL Taq polymerase (BIOTAQ_ DNA Polymerase, 3U/uL), 2.5 uL dNTP (10uM), 2.5

uL amplification buffer (10x), 2.0 uL MgCl, and 16 uwL ddH,O. Cycling parameters

comprised an initial denaturation at 94°C for 5 min, 35 cycles of denaturation at 94°C

for 50 s, annealing at 50-53°C for 50 s, extension at 72°C for 1 min, and a final extension

at 72°C for 5 min.

Allocetraria in China ... 579

TABLE 1. Specimens of Allocetraria spp. and outgroups used in the phylogenetic

analysis

SPECIES

A. ambigua

A. corrugata

A. endochrysea

A. flavonigrescens

*A. flavonigrescens

*A. globulans

A. madreporiformis

A. sinensis

*A. sinensis

A. stracheyi

*A. stracheyi

*A. ambigua

*A. madreporiformis

*Tuckermanopsis ciliaris

*Usnocetraria oakesiana

Vulpicida juniperina

VOUCHER

Wang BM12059

Zhang Z11136

Zhang Z11139

Zhang Z11144

Zhang Z11145

Wang YK12033

Wang BM12011

Wang YK12003

Wang YK12010

Zhang Z11121

Wei & Chen QH12058

Wei & Chen QH12407

Wei & Chen QH12909

Wei & Chen QH12114

Wang BM12062

Obermayer 08140

Obermayer 08137

Wei 125691

Wang YK12008

Wang YK12025

Obermayer 08148

Wang BM12029

Wang YK12005

Wang YK12009

Wang YK12016

Wang YK12030

Wang DQ12432

Obermayer 143

Obermayer 08141

Obermayer 7746

Cao et al. HY11-243

LOCALITY

China

Austria

Germany

China

GENBANK NO.

KF923756

KF923757

KF923758

KF923759

KF923762

KF923760

KF923763

KF923764

KF923765

KF923768

KF923769

KF923770

KF923771

KF923772

KF923773

AF404127

AF404126

KF923774

KF923775

KF923776

AF404125

KF923777

KF923778

KF923779

KF923780

KF923781

KF923782

JX144031

AF404128

AF416460

HQ650615

EU401757

KF923786

* ITS sequences downloaded from GenBank. (All other sequences were obtained by the authors

from specimens deposited in HMAS-L.)

580 ... Wang, Wei, & Wei

DNA sequencing & phylogenetic analysis

PCR products were sequenced using ABI 3700 Sequencer (Shanghai Boshang

Corporation) and analyzed phylogenetically with software Mega5 (Tamura et al. 2011).

The K2+G model was set according to the lowest BIC (Bayesian Information Criterion)

scores. NrDNA ITS sequences of 27 specimens were generated, and 9 sequences were

downloaded from GenBank (TABLE 1). Three genera included in the cetrarioid clade

(Tuckermanopsis ciliaris (Ach.) Gyeln., Usnocetraria oakesiana (Tuck.) M.J. Lai &

J.C. Wei, Vulpicida juniperina (L.) J.-E. Mattsson & M.J. Lai) were used as outgroup. The

phylogenetic tree was inferred by maximum likelihood (ML), and the reliability of the

inferred tree was tested by 1000 bootstrap replications. The intraspecific and interspecific

genetic distances of the Allocetraria species were also calculated and compared.

Results & discussion

Phylogenetic analyses

Eight species of Allocetraria, including the new species, were included in our

phylogenetic analysis. Allocetraria capitata and A. isidiigera are not included

because we were unable to obtain fresh material. The ITS-based ML tree (Fic. 1)

indicates that the 8 Allocetraria species form a well-supported (88% bootstrap

value) monophyletic clade. Each species for which more than one specimen

was included also formed a monophyletic clade. The backbone of the topology

only received moderate support; additional loci will be necessary to address

the phylogenetic relationships within the genus. However, the genetic distances

(TABLE 2) based on nrDNA ITS sequences within Allocetraria provided more

information. According to Del Prado et al. (2010), the distance based on the

nrDNA ITS sequences of parmelioid lichens in Parmeliaceae (one of the largest

lichenized families) has been shown to serve as a powerful tool for identifying

species complexes. In our analysis of Allocetraria, the intraspecific distance

range was 0-0.010 (most <0.006), while the interspecific distance range was

0.014-0.089. The closest distance of A. corrugata was to A. flavonigrescens

(0.028), which was well separated from other species of Allocetraria.

Morphology & chemistry

The Allocetraria thallus is characterized as foliose to subfruticose, dorsiventral

or with radially symmetric branches, dichotomously or subdichotomously

branched; upper surface yellow, greenish yellow, or brown; lobes flat, suberect

to erect; pseudocyphellae angular or sublinear, marginal along the lower

surface; sorediate or isidiate; lower surface wrinkled, yellowish, brown, or black;

sparsely rhizinate; medulla white, yellowish, or orange yellow. Apothecia rare,

terminal; 8 ascospores per asci, simple, near globose, 4-10 um in diameter;

pycnidia marginal, immersed or on emergent projections; pycnoconidia

Allocetraria in China ... 581

@ A. stracheyi KF923782

@ A. stracheyi KF923777

@ A. stracheyi KF923778

*° 1 @ A. stracheyi KF923781

@ A. stracheyi KF923780

@ A. strachey KF923779

64 A. stracheyi JX144031

@ A. ambigua KF923756

A. ambigua AF 404128

g5|@ A. ambigua KF923762

@ A. ambigua KF923759

@ A. ambiguakF923758

@ A. ambigua KF923757

@ A. endochrysea KF923763

5 |@ A. endochrysea KF923764

@ A. endochryseaKF 923765

@ A. sinensis KF923775

IL A. sinensis AF404125

@ A. sinensis KF923776

A. globulans AF 404126

99, A. Madreporiformis AF416460

@ A. madreporiformis KF923774

@ A. comugata KF923760 “| sp. nov.

@ A. flavonigrescens KF923767

: A. flavonigreseens JX144030

@ A. flavonigrescens KF923766

@ A. flavonigrescens KF923772

ka @ A. flavonigrescens KF923770

@ A. flavonigrescens KF923768

@ A. flavonigrescens KF923773

55 | @ A. flavonigrescens KF923771

@ A. flavonigrescens KF923769

A. flavonigrescens AF404127

@ Vulpicida juniperina KF923766

59 Usnocetraria oakesiana EU401757

Tuckermanopsis ciliaris HQ650615

0.01

Fic. 1. The maximum likelihood tree based on nrDNA ITS region sequences of Allocetraria spp.

The samples marked with ‘®’ were examined by the authors. The numbers at each node represent

bootstrap support values =50.

582 ... Wang, Wei, & Wei

TABLE 2. Intraspecific and interspecific genetic distances range of the

Allocetraria species

SPECIES 1 2 3 4 5 6

1 A. ambigua ee

Yin 0.002

2 A. corrugata 0.045 —

0.024— 0.045- 0.002-

A. h

ia aaa 0.026 0.049 + —-0.006

aval favenmevens 0.030- 0.028- 0.032- 0.002-

8 0.035 0.036 0.042 0.010

0.057- 0.069- 0.057-

A. .07

SF oe 0.059 = "7? g073 0.073

0.038- 0.042- 0.026-

A. j j .032 é

6 A. madreporiformis 0.039 0.03 0.047 Ged 0.069 0

oh avn 0.059- 0.057- 0.059- 0.048- 0.079- = -0.059-

: 0.063 0.061 0.067 0.057 0.081 0.063

0.014- 0.047- 0.024— 0.032- 0.040-

8 A. stracheyi 0.065

ae 0.020 0.051 0.030 0.040 0.045

Notes: The numbers in the first row indicate the different species listed in column 1;

— indicates no range because of single sample.

TABLE 3. Diagnostic characters of Allocetraria species

SPECIES SOREDIA/ MEDULLA CONIDIAL CHEMISTRY

ISIDIA LENGTH

A. ambigua Absent White c. 15 um Usnic, lichesterinic,

protolichesterinic,

secalonic acids

A. capitata Capitate Yellowish c.15 um Usnic,

soralia to yellow fumarprotocetraric,

protocetraric, secalonic

acids

A. corrugata Absent White <30 um Usnic,

fumarprotocetraric,

protocetraric acids

A. endochrysea Absent Yellow c. 15 um Usnic, lichesterinic,

protolichesterinic acids,

dufourin, endochrysin

A. flavonigrescens Absent White c. 15 um Usnic,

fumarprotocetraric,

protocetraric acids

A. globulans Absent White c. 15 um Usnic, lichesterinic,

protolichesterinic,

secalonic acids

A. isidiigera Isidiate White c. 15 um Usnic,

fumarprotocetraric,

protocetraric acids

A. madreporiformis Absent White c. 15 um Usnic, lichesterinic acids

A. sinensis Absent White c. 15 um Usnic, lichesterinic,

protolichesterinic,

secalonic acids

A. stracheyi Absent Yellowish = c. 15 um Usnic, lichesterinic,

to yellow protolichesterinic,

secalonic acids

0.002

0.056- — 0-

0.061 0.004

SUBSTRATE

Terricolous,

corticolous

Terricolous

Saxicolous

Terricolous,

corticolous

Terricolous,

corticolous

Terricolous,

corticolous

Corticolous

Terricolous

Terricolous,

saxicolous

Terricolous,

corticolous,

saxicolous

Allocetraria in China ... 583

filiform, 0.5-2x10-25(-30) um. Both cortices are palisade plectenchymatous

or paraplectenchymatous. Usnic acid is present in the cortex, while lichesterinic

and protolichesterinic fatty acids are common in the medulla; secalonic acid

occurs in several, and dufourin, endochrysin, fumarprotocetraric acid or

protocetraric acid in some species.

The new species, A. corrugata, is diagnosed as having long conidia, numerous

black spots on the margin and upper surface, and strongly rugose upper and

lower lobe surfaces. It has the same secondary metabolites as A. isidiigera but

differs in its flat lobes, an obviously rugose upper surface, and longer conidia

(TABLE 3).

All Allocetraria species contain usnic acid, and most contain lichesterinic

and protolichesterinic acids. Allocetraria endochrysea is characterized by

containing dufourin and endochrysin in the medulla. The fumarprotocetraric

acid chemosyndrome occurs in A. corrugata, A. flavonigrescens and A. isidiigera.

Secalonic acid is found in A. ambigua, A. capitata, A. globulans, A. sinensis and

A. stracheyi.

Key to the ten species of Allocetraria in China

L:cThallussubituticose.to-near-columiiary: i. cee foe eh beeen Sa oe aa 4 SES See pe eB a 2

LThallpstoligset.* .asede carpe a tet 3s A Ree ee Mad inn Rohe eee 8 cena S 3

Pe Cro hot aids (og Reenter MALE Mae cia Cub i bk.) Mel et eS CMRI ePieg 22 A A. madreporiformis

ZF MSC IA eNOW: 25 25 Wor ts eeasee st greets ot cepa <2 tees hoes oO Pie 2 oe ts ee A. endochrysea

Dc ISIGIAIOTSOTECHASPROSOIE mb fs fs tat ac ae As Elie eB Morand wae Bhcteerns, coibnne sacepesh pent aces sok nent ae b

Se ESsicinpaniyl SOTedia aDSCHT «4 ot. k tie oA eh gk Doles teckgeree poet Ade hak terete bare oath Ar os eet 5

4. Wwidiarpresent, medullaris. 2 A spMasege opel he esate h inher tek petits ads Stic! A. isidiigera

4 Capitate sordlig: present, tried P D> $a sry yee Such « ree rye Hiker ARE ot Se oe by A. capitata

SPIGA SIDE ctttve 5 ccnebe he, Hae RO Psdgals Dor. gona te tye gs kM oilige oe Botham ht Blidge? A. flavonigrescens

saa Rea b lL Fis 2S em omen SVE Sa, ie, Die ROOKIES MRS Ad ee Ee vs Ra 6

6-Gonidiaup te 25-30 none .o cies tadink ehh cao eeee dod ehee end oleae A. corrugata

G Conicia py LOpl Sup ROO. ek Dae, ccs Ae GR nahn ale nos Ms, Sa nn om, 7

Plsidielike: LObules: present: 40a sy.dye wee oh eee Lats ee oa pe are. ne ee tee epee oe A. stracheyi

Poleidiaclike TOBULeS A Set Nos eo wos sgplh tolt wus ge (ete pect NA eas ak PE cee ol he ete shel es 8

8. leaweranr ace avelowishic of. stews Soka ee hod Pate te ee pe ade bak ee eth Ae A. ambigua

Ss Kower-suttacebrowit(O-DIAGKi xy oe ch. were wore ke Becker ne oes oto ee Ve coeietest eee de pend oe oe 9

STAMOS CESCE Sy Bere a He Ae, op, Banta ee Sab Tien oe ges heal erat At a Rec iee ed A. globulans

9. Tinalliscprocuimbente. «405 5 aK karte cs Bet oh ae Rima PEAS aE 0 eel rattle te A. sinensis

584 ... Wang, Wei, & Wei

FERRERS ER RCRA SSR RENEE R TERRE PEER R RRR e eee

4 :

oe | ce] D

= Vi

7 TRRURRGRRROR TERRA ERROR ERE EE, ii ii iii iiiiiil —EEO—e=>E—vz——see

>» ‘tp x pay. |

"% >

PLATE 1. Habit of Allocetraria species. A. A. ambigua (HMAS-L 015005); B. A. capitata (isotype);

C. A. endochrysea (HMAS-L 127390); D. A. isidiigera (holotype); E. A. madreporiformis

(HMAS-L125691); F. A. sinensis (HMAS-L 127405); G. A. stracheyi (HMAS-L 015149);

H. A. flavonigrescens (HMAS-L127395); I. A. flavonigrescens (HMAS-L 069642); J. A. globulans

(HMAS-L015177). Scale in mm.

Allocetraria in China ... 585

Species

Allocetraria ambigua (C. Bab.) Kurok. & M.J. Lai, Bull. Natl. Sci. Mus.,

Tokyo, B 17: 62, 1991. PLATE 1A

Characterized by thin flat channeled lobes and a yellowish lower surface,

A. ambigua is similar to A. stracheyi, which is distinguished by thick convex

lobes and isidia-like lobules.

CHEMISTRY— Usnic, lichesterinic, protolichesterinic, and secalonic acids.

SUBSTRATE—On soil or bark.

DISTRIBUTION—China (Gansu, Shaanxi, Sichuan, Tibet and Yunnan

provinces), India, Nepal.

SPECIMENS EXAMINED: CHINA: Gansu PRov.: Mt. Minshan, alt. 4200 m, 23 August

1937, T.P. Wang 7582 (HMAS-L 014999); SHAANXI PRov.: Mt. Taibaishan, alt. 3200-

3700 m, 12-13 July 1988, X.Q. Gao 3158 (HMAS-L 015003); X.Q. Gao 3185 (HMAS-L

015004); C.H. Ma 236 (HMAS-L 015000); C.H. Ma 238 (HMAS-L 015002); C.H. Ma

375 (HMAS-L 015005); alt. 3610 m, 23 July 2011, T. Zhang Z11136 (HMAS-L 127368);

T. Zhang Z11139 (HMAS-L 127370); T. Zhang Z11144 (HMAS-L 127367); T. Zhang

Z11145 (HMAS-L 127369); SICHUAN PRov.: Mt. Gonggashan, alt. 4200 m, 29 July 1982,

X.Y. Wang et al. 9046 (HMAS-L 015009); TrsET: Nyalam County, 12 June 1966, J.C. Wei

& J.B. Chen 1441 (HMAS-L 015137); YUNNAN PrRov.: Degen County, Mt. Baibaxueshan,

alt. 4560 m, 16 September 2012, R.E Wang BM12059 (HMAS-L 127372).

Allocetraria capitata R.F. Wang, Li S. Wang & J.C. Wei, Mycosystema 33: 21, 2014.

PLATE 1B

Characterized by having capitate soralia, A. capitata is similar to A. isidiigera,

which is distinguished by its absence of soredia and presence of isidia.

CHEMISTRY— Usnic, fumarprotocetraric, protocetraric, and secalonic acids.

SUBSTRATE—On soil, often over moss.

DIsTRIBUTION—China (Sichuan Province).

SPECIMENS EXAMINED: CHINA: SICHUAN PRov.: Dege Country, Manigangge villages,

Mt Queershan, 31°55’N 98°56’E, alt. 4510 m, on moss, 30 August 2007, L.S. Wang et al.

07-28259 (KUN—holotype; HMAS-L—isotype).

Allocetraria corrugata R.F. Wang, X.L. Wei & J.C. Wei, sp. nov. PLATE 2

FUNGAL NAME FN 570087

Differs from Allocetraria isidiigera by its longer conidia, numerous black spots on the

margin and upper surface, and strongly rugose upper and lower lobe surfaces.

TypE—China, Yunnan Prov., Deqin Co., Meli village, Meili snow mountain, on rock,

28°38’N, 98°37’E, alt. 4400 m, 10 Sep. 2012, R.E Wang 12033 (Holotype, HMAS-L).

EryMoLoGy—tThe epithet corrugata refers to the corrugated lobe surfaces.

Thallus foliose, adnate, dorsiventral; lobes narrow, 1-4 mm wide, slightly

inflated, 200-450 um thick, sublinear-elongate, sub-dichotomous to irregular

branches, lobules irregularly branched; upper surface greenish yellow to

586 ... Wang, Wei, & Wei

PLATE 2. Allocetraria corrugata (holotype). A. Habit; B. The numerous black spots on the margin

and upper surface; C. Longitudinal section of the thallus; D. Conidia; E. Longitudinal section of

the thallus with black spots, hyphae inside and the arrow pointing at the location of one black spot;

E Magnification of the black spot pointed by the arrow in E. Scale bars: A = 1 cm; B= 1 mm; C, D

= 20 um; E, F=5 um.

Allocetraria in China ... 587

green, strongly rugose, dark brown and whitish pruinose at the apices of

lobes; spots dark brown to black, abundant, flush to emergent, scattered to

crowded, mostly covered by white pruina; lower surface gray-white to pale

brown, conspicuously rugose, rhizines marginal and laminal, concolorous

with lower surface, sparse, unbranched, 1-4 mm long; pseudocyphellae rare,

white, located on the ridges of the underside or on outgrowths; medulla white.

Upper cortex indistinct palisade plectenchymatous, 20-30 um

thick, composed of 3-6 layers; algal layer 25-50 um; lower cortex

paraplectenchymatous, 15-20 um thick, composed of 3-4 layers; black spots

at the upper surface not separated, composing of hyphae, the algal layer

continuous. Apothecia not seen. Pycnidia frequent, both immersed and

elevated on black projection, also at the top of isidia-like structures on the

upper surface and margin, conidia colorless, filiform, one end slightly swollen,

(12.5-)15-25(-30)x0.5-1.5 um.

CHEMISTRY—Cortex: K-, C-, KC+ yellow, PD-; medulla: K-, C-, KC-,

PD+ yellow to red; containing usnic, fumarprotocetraric, and protocetraric

acids (TLC).

SUBSTRATE—On rock, overgrowing mosses.

DisTRIBUTION—Currently known only from Yunnan Province, China.

Allocetraria endochrysea (Lynge) Karnefelt & A. Thell, Nova Hedwigia 62: 507,

1996. PLATE 1C

Characterized bya subfruticose thallus with a yellow medulla, A. endochrysea

is similar to A. madreporiformis, which is readily distinguished by its less-

branched lobes, white medulla, and absence of dufourin and endochrysin.

CHEMISTRY— Usnic, lichesterinic, and protolichesterinic acids, dufourin,

endochrysin.

SUBSTRATE—On soil, overgrowing mosses.

DIsTRIBUTION—China (Sichuan and Yunnan provinces).

SPECIMENS EXAMINED: CHINA: SICHUAN PRov.: Mt. Gonggashan, alt. 4400 m, 21

October 1999, L.H. Chen 990051 (HMAS-L 127380); L.H. Chen 990087-1 (HMAS-L

127382); YUNNAN PrRov.: Degen County, Mt. Baibaxueshan, alt. 4350-4440 m, 16

September 2012, R.R Wang BM12011 (HMAS-L 127385); R.E Wang BM12040

(HMAS-L 127388); 22 September 2012, Q.M. Zhou et al. DQ12432-1 (HMAS-L

127387); alt. 4800 m, 10 September 2012, R.E Wang YK12001 (HMAS-L 127384); R.F.

Wang YK12003 (HMAS-L 127391); RE Wang YK12010 (HMAS-L 127390); R.E Wang

YK12013 (HMAS-L 127389).

Allocetraria flavonigrescens A. Thell & Randlane, Flechten Follmann Contr. Lich.:

3593. 1995. PLATE 1H, I

Characterized by long narrow channeled lobes, white medulla, and absence

of pseudocyphellae, A. flavonigrescens is similar to A. isidiigera, which is

distinguished by the presence of isidia.

588 ... Wang, Wei, & Wei

CHEMISTRY— Usnic, fumarprotocetraric, and protocetraric acids.

SUBSTRATE—On soil and corticolous on branches of Rhododendron.

DISTRIBUTION—China (Qinghai, Shaanxi, Sichuan, Tibet, and Yunnan

provinces), Nepal.

SPECIMENS EXAMINED: CHINA: QINGHAI PRov.: Maqin County, alt. 3970 m, 11 August

2012, X.L. Wei & K. Chen QH12058 (HMAS-L127395); X.L. Wei & K. Chen QH12407

(HMAS-L 128221); X.L. Wei & K. Chen QH12909 (HMAS-L 127447); Banma County,

alt. 4299 m, 4 August 2012, X.L. Wei & K. Chen QH121141 (HMAS-L127394); SHAANXI

Prov.: Mt. Taibaishan, alt. 3200 m, 12 July 1988, C.H. Ma 146 (HMAS-L 015094); alt.

3300 m, X.Q. Gao 3059 (HMAS-L 015091); X.Q. Gao 3051 (HMAS-L 015089); X.Q.

Gao 3055 (HMAS-L 015092); alt. 3480 m, 23 July 2011, T. Zhang Z11121 (HMAS-L

128223); alt. 3360 m, 4 August 2005, J. Yang YJ218 (HMAS-L 069642); SICHUAN PRov.:

Mt. Zheduoshan, alt. 4200 m, 27 September 2007, L.S. Wang 07-28991 (KUN); TIBET:

Mt. Dongdalashan, alt. 4950 m, HMAS-L 032363; YUNNAN PRov.: Mt. Baibaxueshan,

alt. 4560 m, R.E Wang BM12062 (HMAS-L127396); R.E Wang YK12036 (HMAS-L

126786); R.E Wang YK12060 (HMAS-L 128222).

Allocetraria globulans (Nyl.) A. Thell & Randlane, Flechten Follmann Contr. Lich.:

360, 1995. PLATE 1J

A corticolous species characterized by long narrow channeled flat lobes

with corrugated margins, A. globulans is similar to A. stracheyi, which is

distinguished by convex lobes.

CHEMISTRY— Usnic, lichesterinic, protolichesterinic, and secalonic acids.

SUBSTRATE—On bark.

DIsTRIBUTION—China (Yunnan Province), Nepal.

SPECIMENS EXAMINED: CHINA: YUNNAN PRov.: Mt. Yulongxueshan, alt. 3820 m, 6

August 1981, X.Y. Wang et al. 6854 (HMAS-L015180); alt. 3380 m, 6 August 1981, X.Y.

Wang et al. 6550 (HMAS-L015179); alt. 3950 m, 6 August 1981, X.Y. Wang et al. 5110

(HMAS-L015178); Mt. Baimaxueshan, alt. 2700 m, 12 July 1981, X.Y. Wang et al. 7781

(HMAS-L015177).

Allocetraria isidiigera Kurok. & M. J. Lai, Bull. Natl. Sci. Mus.,

Tokyo, B 17: 62, 1991. PLATE 1D

Characterized by long narrow channeled lobes, a white medulla, and the

absence of pseudocyphellae, A. isidiigera is similar to A. flavonigrescens, which

is distinguished by the presence of isidia and slightly smaller conidia.

CHEMISTRY— Usnic, fumarprotocetraric, and protocetraric acids.

SUBSTRATE—Corticolous on Rhododendron branches.

DIsTRIBUTION—China (Tibet), Nepal.

SPECIMENS EXAMINED: CHINA: XIZANG (TIBET): Nyalam County, on Rhododendron

stem, alt. 3910 m, J.C. Wei & J.B. Chen 1857 (Holotype, HMAS-L).

Allocetraria in China ... 589

Allocetraria madreporiformis (Ach.) Karnefelt & A. Thell, Nova Hedwigia 62: 508,

1996. PLATE 1E

Characterized by a subfruticose thallus, erect lobes, and a white medulla,

A. madreporiformis is similar to A. endochrysea, which is distinguished by a

yellow medulla and the presence of dufourin and endochrysin.

CHEMISTRY— Usnic and lichesterinic acids.

SUBSTRATE—On soil.

DiIsTRIBUTION—China (Xinjiang Uygur Autonomous Region), Mongolia,

Turkistan, West of North America, Central Europe.

SPECIMENS EXAMINED: CHINA: XINJIANG UyGUR AUTONOMOUS REGION: Mt.

Tianshan, alt. 3200 m, 20 July 1978, X.Y. Wang 0950 (HMAS-L006687); 28 May 2011, J.C.

Wei s.n. (HMAS-L125691); alt. 3000 m, 3 July 1997, X.Y. Wang 367 (HMAS-L006685).

Allocetraria sinensis X.Q. Gao, Flechten Follmann Contr. Lich.: 365, 1995. PLATE 1F

Characterized by an erect thallus, long narrow channeled lobes, and a glossy

lower surface, A. sinensis resembles A. ambigua, which can be distinguished by

its yellowish lower surface.

CHEMISTRY— Usnic, lichesterinic, protolichesterinic, and secalonic acids.

SUBSTRATE—On soil.

DISTRIBUTION—China (Shaanxi, Sichuan, Yunnan, and Tibet), Nepal.

SPECIMENS EXAMINED: CHINA: SHAANXI PRov.: Mt. Taibaishan, alt. 3400 m, on

ground, X.Q. Gao 3052 (Holotype, HMAS-L); alt. 3550 m, 13 July 1988, X.Q. Gao

3178 (HMAS-L 014992); SICHUAN PRov.: Mt. Gonggashan, alt. 2850 m, 24 June 1982,

X.Y. Wang et al. 8260 (HMAS-014986); Mt. Balangshan, alt. 4300 m, 18 August 1982,

X.Y. Wang et al. 9544 (HMAS-014990); X.Y. Wang et al. 9496 (HMAS-L 014988);

TIBET: Zuogong County, alt. 4500 m, 8 October 1982, J.J. Su 5381 (HMAS-L014995);

YUNNAN Prov.: Mt. Yulongxueshan, alt. 3970 m, 6 August 1981, X.Y. Wang et al. 7076

(HMAS-L 014996); Mt. Baimaxueshan, 16 September 2012, R.E Wang YK12006

(HMAS-L 127405); R.R. Wang YK12008 (HMAS-L 127410); R.E. Wang YK12025

(HMAS-L 127406).

Allocetraria stracheyi (C. Bab.) Kurok. & MJ. Lai, Bull. Natl. Sci. Mus.,

Tokyo, B 17: 62, 1991. PLATE 1G

Characterized by thick raised lobes and a yellowish lower surface, A.

stracheyi is similar to A. ambigua, which differs in its thin flat lobes, and A.

capitata, which differs in the presence of soredia.

Chemistry.—Usnic, lichesterinic, protolichesterinic, and secalonic acids.

SUBSTRATE—On soil and branches of shrubs.

DIsTRIBUTION—China (Shaanxi, Sichuan, Xinjiang Uygur Autonomous

Region, Yunnan, and Tibet), India, Nepal, North America.

SPECIMENS EXAMINED: CHINA: SHAANXI Prov.: Mt. Taibaishan, alt. 3600 m, 13 July 1988,

X.Q. Gao 3151 (HMAS-L 015174); StcHuAN PrRov.: Mt. Gonggashan, alt. 3700-3800 m,

590 ... Wang, Wei, & Wei

27 June 1982, X.Y. Wang et al. 8354 (HMAS-L 015161); Mt. Balangshan, alt. 4300 m, 18

August 1982, X.Y. Wang et al. 9488 (HMAS-L 015168); TrBET: Zuogong County, alt. 4400

m, 8 October 1982, J.J. Su 5387 (HMAS-L015155); YUNNAN PrRov.: Mt. Baimangxueshan,

alt. 4300 m, 29 August 1981, X.Y. Wang et al. 7546 (HMAS-L 029482); Mt. Daxueshan,

alt. 4250 m, 8 September 1981, X.Y. Wang 7209 (HMAS-L015154); alt. 4330-4800 m,

8 August 1982, J.J. Su 5410 (HMAS-L 015149); Degen County, 16 September 2012,

R.E Wang BM12029 (HMAS-L 127423); R.E Wang YK12005 (HMAS-L 127428);

R.E Wang YK12009 (HMAS-L 127418); R.E Wang YK12016 (HMAS-L 127427);

R.E Wang YK12030 (HMAS-L 127430); RE Wang DQ12432 (HMAS-L 127424);

XINJIANG UyGuR AUTONOMOUS REGION: Mt. Tianshan, alt. 3200 m, 20 July 1978, X.Y.

Wang 949 (HMAS-L 007697).

Acknowledgements

We are indebted to Prof. H. Thorsten Lumbsch and Prof. Jae Seoun Hur for giving

valuable comments on the manuscript. This project was supported by the National

Natural Science Foundation of China (31200018) and Main Direction Program of

Knowledge Innovation of Chinese Academy of Sciences (KSCX2-EW-Z-9). We thank

Ms. H. Deng for giving considerable assistance during the studies in HMAS-L.

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Culberson CF, Kristinsson H. 1970. A standardized method for the identification of lichen products.

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Del Prado R, Cubas P, Lumbsch HT, Divakar PK, Blanco O, Amo De Paz G, Molina MC, Crespo

A. 2010. Genetic distances within and among species in monophyletic lineages of Parmeliaceae

(Ascomycota) as a tool for taxon delimitation. Molecular Phylogenetics and Evolution 56:

125-133. http://dx.doi.org/10.1016/j.ympev.2010.04.014

Karnefelt I, Thell A. 1996. A new classification for the Dactylina/Dufourea complex. Nova Hedwigia

62: 487-511.

Karnefelt I, Thell A, Randlane T, Saag A. 1994. The genus Flavocetraria Karnef. & ‘Thell

(Parmeliaceae) and its affinities. Acta Botanica Fennica 150: 79-86.

Kurokawa S, Lai MJ. 1991. Allocetraria, a new genus in the Parmeliaceae. Bulletin of the National

Sciences Museum, Tokyo, B 17: 59-65.

Lai MJ, Qian ZG, Xu L. 2007. Synopsis of the cetrarioid lichen genera and species (Parmeliaceae,

lichenized Ascomycotina) in China. Journal of the National Taiwan Museum 60: 45-61.

Nelsen MP, Chavez N, Sackett-Hermann E, Thell A, Randlane T, Divakar PK, Rico VJ, Lumbsch,

HT. 2011. The cetrarioid core group revisited (Lecanorales: Parmeliaceae). Lichenologist 43:

537-551. http://dx.doi.org/10.1017/S0024282911000508

Randlane T, Saag A. 1992. New combinations of some cetrarioid lichens (Parmeliaceae). Mycotaxon

44; 491-493.

Rogers SO, Bendich AJ. 1988. Extraction of DNA from plant tissues. 1-10, in: SB Gelvin, RA

Schilperoort (eds). Plant Molecular Biology Manual, vol. A6. Kluwer Academic Publishers,

Boston.

Saag A, Randlane T, Thell A, Obermayer W. 2002. Phylogenetic analysis of cetrarioid lichens with

globose ascospores. Proceedings of the Estonian Academy of Sciences, Biology, Ecology 51:

103-123.

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Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: Molecular

evolutionary genetics analysis using maximum likelihood, evolutionary distance, and

maximum parsimony methods. Molecular Biology and Evolution 28: 2731-2739.

http://dx.doi.org/10.1093/molbev/msr121

Thell A, Randlane T, Karnefelt I, Gao XQ, Saag A. 1995. The lichen genus Allocetraria (Ascomycotina,

Parmeliaceae). 353-370, in: FJ Daniels et al. (eds). Flechten Follmann. Contributions to

Lichenology in Honour of Gerhard Follmann, University of Cologne, Germany.

Thell A, Hégnabba F, Elix JA, Feuerer T, Karnefelt 1, Myllys L, Randlane T, Saag A, Stenroos S,

Ahti T, Seaward MRD. 2009. Phylogeny of the cetrarioid core (Parmeliaceae) based on five

genetic markers. Lichenologist 41: 489-511. http://dx.doi.org/10.1017/S0024282909990090

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238-4246.

Wang RF, Wang LS, Wei JC. 2014. Allocetraria capitata sp. nov. (Parmeliaceae, Ascomycota) from

China. Mycosystema 33: 19-22. http://dx.doi.org/10.13346/j.mycosystema.130084

White TJ, Bruns T, Lee S, Taylor JW. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR protocols: a guide to

methods and applications. Academic Press, San Diego, California.

MYCOTAXON

http://dx.doi.org/10.5248/130.593

Volume 130, pp. 593-598 April-June 2015

Book reviews and notices

ELSE C. VELLINGA (GUEST BOOK REVIEW EDITOR)

"861 Keeler Avenue, Berkeley CA 94708 U.S.A.

" CORRESPONDENCE TO: bookreviews@mycotaxon.com

ABSTRACT—Books reviewed include: Ascomycota—Catalogue of discomycetes referred

to the genera Helotium Pers. and Hymenoscyphus Gray (Lizon & Kucera 2014);

Basidiomycota—FunkKey: an interactive guide to the macrofungi of Australia. Key to

agarics (May & al. 2014); CHECKLIstT—Kommentierter Katalog der Flechtenbewohnenden

Pilze Bayerns (von Brackel 2014).

Two different types of checklists are reviewed in this instalment of the book

reviews. The first, which is of nomenclatorial interest, deals with the genera

Helotium and Hymenoscyphus; the second checklist lists the fungi living

on lichens in Bavaria. The third review looks at an interactive key for the

genera of agarics in Australia. I foresee more of the latter type of publication

in the future!

ASCOMYCOTA

Catalogue of Discomycetes referred to the genera Helotium Pers. and

Hymenoscyphus Gray. By P. Lizon & V. Kucera. 2014. Institute of Botany,

Slovak Academy of Sciences, Dubravska cesta 9, SK-84523 Bratislava, Slovakia.

<www.ibot.sav.sk> ISBN 978-80-971837-4-5. 144 pp. Price not known.

Since 1801 (or at least from 1849 onwards), more or less stalked, non-hairy

and non-gelatinous inoperculate discomycetes were generally classified in the

* Book reviews or books for consideration for coverage in this column should be sent to the

address above OR to the Editor-in-Chief at editor@mycotaxon.com or 6720 NW Skyline,

Portland OR 97229 USA.

594 ... Vellinga (editor)

genus Helotium Pers., essentially paving the way for the genus to become an

enormous heterogeneous collection of taxa. When in 1962 R.W.G. Dennis

argued that this genus should be split up into a (small) genus Helotium and a

(large) genus Hymenoscyphus Gray, M.A. Donk demonstrated that Helotium

Pers. was a later homonym of the basidiomycete generic name Helotium Tode,

thereby soon ending the currency of the discomycete genus Helotium. Since

then, the genus Hymenoscyphus has increased considerably, by both new

combinations and newly described taxa. It is beyond doubt that mycologists

working on this group cannot avoid taxa once classified in Helotium and/or

Hymenoscyphus, but they must make their way without world monographs of

these species-rich genera.

Therefore Lizon & Kucera have filled a gap by making a catalogue (checklist)

of no less than 1183 specific and infraspecific epithets known to have at one

time been attributed to Helotium and/or Hymenoscyphus. This number includes

all validly published and a few unpublished or invalidly published names

used in literature. Each entry comprises the Helotium and/or Hymenoscyphus

name, the authority, the basionym, data about type substrate, country and

specimen(s), the current name according to the authors or other authorities,

and important taxonomic and/or nomenclatural remarks (e.g., about

nomina nova or superfluous names). One entry does not necessarily present

the complete synonymy of the taxon concerned. Where there is insufficient

taxonomic information available or where there are different taxonomical

opinions, a bold-faced current name is omitted. A short introductory chapter

on the taxonomy and nomenclature of both genera and their common order

(Helotiales) and family (Helotiaceae) precedes the actual catalogue. The book

ends with an index of additional epithets mentioned within the entries.

It is evident that this publication has brought together a huge amount of

relevant nomenclatural and taxonomical information about the taxa within

the genera concerned. It also makes clear which taxa are, as yet, insufficiently

known well enough for accurate classification.

Some critical remarks have certainly to be made here. In addition to many

mistakes in spelling, typography (boldfaced instead of regular font), and

grammar, the catalogue contains numerous inconsistencies. Abbreviations of

author names, books, and periodicals are said to follow ‘generally’ (why not

always?) the well-known standards, but the number of exceptions is fairly large:

e.g., always “Thind’ instead of ‘K.S. Thind’ and sometimes ‘Buchwald’ for ‘N.E.

Buchw, “W. Phill? for “W. Phillips; “Ell? for “Ellis; “Sharma’ or ‘P.M. Sharma’ for

‘M.P. Sharma, ‘B. Declercq’ for “Declerq; ‘J. Hengstm’ for “Hengstm.,, ‘P. Karsten’

for ‘P. Karst., ‘Monogr. Discomyc. Boh? (Velenovsky) for “Monogr. Discomyc.

Book Reviews ... 595

Bohem,, and ‘Bidr. Kanned. Finl. Nat. Folk’ for ‘Bidrag Kannedom Finlands

Natur Folk’ etc.

The way in which the authors use the equality sign (=), indicating

heterotypic synonymy, and the triple bar (=), indicating homotypic synonymy,

is disputable. Combinations of Helotium or Hymenoscyphus and the epithet

of an entry are not preceded by such a triple bar. If the basionym concerned

or the current name, if homotypic, does not belong to one of these genera,

it is preceded by a triple bar. However, sometimes the basionym is wrongly

preceded by an equality sign (e.g., Allophyllaria filicum) or by no sign at all

(e.g., Phialea bicolor). A nomen novum or recombination that is homotypic

with the replaced name should be preceded by a triple bar, at any rate not by

an equality sign as mostly is done in this book (e.g., under destructor, robergei,

and vernale). It would have been more logical to precede not only the basionym

but all its recombinations and nomina nova by a triple bar, as is usually done.

As far as possible, in each entry the current name is indicated and printed

in bold (see above). Unfortunately this has been done rather inconsistently. For

example, the entries imberbe and pileatum (P. Karst.) each contain two ‘current’

names. Under cupularum, Hym. imberbis is mentioned as the current name,

but under obliquum its homotypic synonym, Phaeohelotium imberbe, is flagged

as current. In several entries a current name is indicated which, according

to the entry of the epithet concerned, is Not the current name. For example,

under hymenulus and ostruthii, Hym. herbarum is mentioned as current name,

while under herbarum it is stated that Calycina herbarum is the current name.

Similarly this applies to Hym. epiphyllus in the entries gyalectoides and lunatum

resp. epiphyllum. Under phiala, an invalidly published name is even marked as

the ‘current’ name.

The authors appear to have paid attention to the inclusion of orthographic

errors that, as usual, are placed between single quotation marks and brackets

behind the corrected form. However, many orthographic errors remain, even

in the labels of the entries, e.g., aspidiicolum, caulicolum, and pinicolum.

The conclusion may be that this catalogue is certainly an asset for all

mycologists who are engaged with helotioid discomycetes. Unfortunately the

many inaccuracies, which could have been prevented by a more thorough final

editing, detract from the quality. Therefore this catalogue is not the dependable

standard work for both genera that it might have been.

JAN HENGSTMENGEL

Naturalis Biodiversity Center, P.O. Box 9517

2300 RA Leiden, The Netherlands

j.hengstmengel@zonnet.nl

596 ... Vellinga (editor)

BASIDIOMYCOTA

FunKey: an Interactive Guide to the Macrofungi of Australia. Key to Agarics.

version 1. By T. W. May, K. Thiele, C. W. Dunk, S. H. Lewis. 2014. Identic,

Brisbane and ABRS, Canberra. <http://shop.lucidcentral.org/>. ISBN 978-0-642-

56877-9. 1200+ images (numerous in colour) for 159 taxa and 383 character states.

Price: $50.73 (USB 2.0 Key).

FunKey is also available as an “app” for mobile devices using Apple or Android

systems. The app provides the interactive key and fact sheets for genera and

characters. However, the app does not include the extensive introductory material

on genera and characters or the glossary that is present on the USB version. In the

app, the best function is available, but not other Lucid key functions such as subsets

or ranked sort. Purchase direct from either the iTunes store:

https://itunes.apple.com/WebObjects/MZStore.woa/wa/viewS oftware? id=958085767&mt=8

or Google Play:

https://play.google.com/store/apps/details?id=com.lucidcentral.mobile.funkey&hl=en

According to its introduction, “FunKey: Key to Agarics” is an interactive key

and information system for the agaric genera occurring in Australia.

FunKey is available on a USB key or, in a slimmed down version, as an app for

mobile devices. I reviewed the USB version. The system requirements for this

program are an Internet browser with installed Java plug-ins. Once installed,

it feels like working on-line, even though all data are present on the USB key.

The main part is formed by the interactive keys where you can start with

whatever character you wish; as you select a character from a list of characters

and character states, groups of taxa or genera are left in the in-group while

others move to the discarded group. For instance, if you choose white spores,

all species with coloured spores are immediately eliminated from the list of

possible candidates. The final selections can be further examined, as the fact-

sheets for every species/species group provide extensive information.

The keys work well; I tested them with Macrolepiota clelandii and an

Agaricus. However, users should be well aware that in many cases the keys do

not lead you to a species but to a genus, species group, or section of a genus. A

second caveat is that for many Australian species, names are not yet available,

so that can be frustrating as well.

What makes this so valuable is all the background information. There is a

fact sheet for each genus or group with notes on characteristics, distribution

in Australia, photos, a list of references; there is also extensive discussion and

argument for generic placements, a glossary for all the terms used in the keys

and descriptions, and a compiled list with all references.

Book Reviews ... 597

These resources make this a very useful work for people in other countries.

FunKey is also an example how to make data on fungi more easily accessible

than is possible in traditional publishing formats such as dichotomous keys or

written descriptions without keys. Of course, as version 1 is only the beginning,

Iam looking forward to expanded updates with keys to all the named species.

CHECKLIST

Kommentierter Katalog der Flechtenbewohnenden Pilze Bayerns. By W. von

Brackel, 2014. BrisLiIoTHECA LICHENOLOGICA Volume 109. J. Cramer, Johannesstr.

3A, 70176 Stuttgart, Germany, mail@schweizerbart.de, <www.borntraeger-cramer.

de> ISBN 1436-1698. 476 pages, 13 illustrations. Price 119.00 €

Since 2004, von Brackel has focused on the lichenicolous fungi of Bavaria, of

which he has documented and identified 1900 collections. The present work

is the result of that very labour intensive research, conducted mostly in the

author's evenings and weekends.

In all, von Brackel has identified (and here reports on) 403 species in 141

genera, including 372 obligate lichenicolous species and 31 others that are

partly lichenized, facultatively lichenicolous, or have a taxonomy and ecology

that are not well known. Five species new for science are introduced, and one

new combination is proposed.

The book starts with a short introduction outlining the history of the

discovery and cataloguing of the lichenicolous fungi in Bavaria—a history that

goes back to the early 1800s. The author emphasizes that wherever there are

lichens, there are fungi growing on them. However, it is important to remember

that the ecological requirements of host and fungal symbiont are not always the

same, and that their sensitivity to air pollution might differ.

The methods section explains how the data were collected in the field, where

the collections are conserved, and how the book is organized.

The most important part follows: an alphabetical enumeration of genera

and species that is over 400 pages long. For each genus a short introduction

on taxonomic position, morphological characters, and ecology is given. The

species are treated more in depth: name, synonyms, hosts, literature, comments,

distribution (based on literature), distribution in Germany, finds in Bavaria

(literature), and the author’s own observations and collections are enumerated.

Acknowledgements followed by a list of references (almost 60 pages long)

complete this work.

598 ... Vellinga (editor)

The ‘catalogue’ is a fascinating and important contribution to our

knowledge of an often-neglected group of fungi. With this overview for

Bavaria, comparisons with the checklist of lichenicolous fungi in Great Britain

and Ireland (Hawksworth 2003) can be made, leading to better insight in

distribution patterns and ecology.

Iam nota specialist in this group, so I cannot comment on the nomenclature,

and nomenclatorial issues always arise while compiling a book like this. While

browsing through its pages, however, I noticed only one typographical error.

The author can be congratulated on such an important and thorough work.

I hope that this printed book will also be available in an electronic version,

which would make it so much easier to update and to consult than the hard-

copy edition.

Hawksworth DL, 2003. The lichenicolous fungi of Great Britain and Ireland: an overview

and annotated checklist. The Lichenologist 35: 191-232.

Book ANNOUNCEMENTS

Hypocrealean lineages of industrial and phytopathological importance. By

L. Lombard, J.Z. Groenewald & PW. Crous (Eds). 2015. Srupizs in Mycotoey 80.

<www.cbs.knaw.nl>. ISSN 0166-0616. 245 pp., full colour. Price: € 70.00(free download).

Philatelic Mycology: Families of fungi. By W.EO. Marasas, H.M. Marasas,

M.J. Wingfield & P.W. Crous. 2014. CBS BioprversiTy SERIES 14. <www.cbs.knaw.nl>.

ISBN 978-90-70351-99-1. 107 pp., full colour. Price: € 40.00.

MY COTAXON

http://dx.doi.org/10.5248/130.599

Volume 130, pp. 599-600 April-June 2015

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 130(2)

Allocetraria corrugata R.F. Wang, X.L. Wei & J.C. Wei, p. 585

Anaexserticlava T.S. Santa Izabel, R.E Castafieda & Gusmao, p. 446

Anaexserticlava caatingae T.S. Santa Izabel, R.E Castahfeda & Gusmao, p. 446

Aspicilia volcanica Ismayil, A. Abbas & S. Y. Guo, p. 545

Brachycephala J.S. Monteiro, Gusmao & R.E. Castaneda, p. 490

Brachycephala exotica J.S. Monteiro, Gusmao & R.E. Castaneda, p. 490

Chaetochalara mutabilis C.R. Silva, S.S. Silva, Gusmao & R.E Castaneda, p. 506

Corneromyces murrillii (Burt) Nakasone, p. 374

Cryptocoryneum parvulum S.S. Silva, Gusmao & R.F. Castafieda, p. 466

Digicatenosporium S.M. Leao, Gusmao & R.E. Castaneda, p. 480

Digicatenosporium polyramosum S.M. Leao, Gusmao & R.F. Castaneda, p. 480

Diplococcium heteroseptatum D.A.C. Almeida, T.S. Santa Izabel, R.E Castafeda &

Gusmao, p. 497

Distophragmia R.F. Castaneda, $.M. Leao & Gusmao, p. 499

Distophragmia rigidiuscula (R.F. Castafieda) R.E. Castafieda, S.M. Leao &

Gusmao, p. 500

Duportella lassa Spirin & Kout, p. 484

Endophragmiella selenosporellaria Y.R. Ma & X.G. Zhang, p. 452

Graphilbum tsugae J. Reid & Georg Hausner, p. 409

Graphis hongkongensis Wei Guo & J.S. Hur, p. 431

Leptogium taibaiense H.J. Liu & M.Q. Xi, p. 472

Leptogium wangii H.J. Liu & J.S. Hu, p. 473

Leucoagaricus lahorensis Qasim, Amir & Nawaz, p. 534

Matsushimiella paraensis J.S. Monteiro, R.F. Castafieda & Gusmao, p. 312

Melanoderma disciforme H.S. Yuan, p. 424

Micropsalliota pseudoglobocystis Li Wei & R.L. Zhao, p. 558

Neosporidesmium diaoluoshanense Xiang Y. Li & X.G. Zhang, p. 308

600 ... MyCcoTAXON 130(2)

Phaeoschizotrichum C.R. Silva, Gusmao & R.E. Castaneda, p. 438

Phaeoschizotrichum ramosum C.R. Silva, Gusmao & R.E. Castaneda, p. 438

Polyporus epitheloides Nakasone, p. 388

= Peziza flava Sw. : Fr., 1788

non Polyporus flavus Jungh., 1838

Polyporus polyacanthophorus Nakasone, p. 383

= Hydnum brunneoleucum Berk. & M.A. Curtis, 1857,

non Polyporus brunneoleucus Berk., 1846

Pseudolagarobasidium conspicuum (Pouzar) Nakasone, p. 378

Pyriculariopsis bicolorata C.R. Silva, Gusmao & R.E. Castaneda, p. 441

Rosellinia camphorae Q.R. Li, J.C, Kang & K.D. Hyde, p. 565

Rosellinia sigmoidea Q.R. Li, J.C, Kang & K.D. Hyde, p. 564

Spadicoides sylvatica Heredia, R.F. Castafeda & R.M. Arias, p. 512

Sporidesmiella mammillata Heredia, R.F. Castafeda & R.M. Arias, p. 514

Vararia amurensis (Parmasto) Nakasone, p. 377