Iv ... MYCOTAXON 130(1)

MY COTAXON

VOLUME ONE HUNDRED THIRTY (1) — TABLE OF CONTENTS

COVER SECTION

EPA inch on okt il sna eRe Meebo oy Aicch OORT AL Sey Dente cA Mea ina At eae RR apna vale 4 vi

ROVICWICTS ENE ry oh OE eee cE EEE Rh Eth Ae hee ee EAS Oke ete es vii

SOI TSSIOMDTOCCHUMES 3 25 vi Peurizh date ths Red ng er a Ma meri decease 8 Ie Viii

ERO TE PEEL Mosty BF Soy inn Pie de On ay wan, A hey nme MERE ix

RESEARCH ARTICLES

Pseudoplectania lignicola sp. nov. described from central Europe

S. GLEJDURA, V. KUCERA, P. LIZON, & V. KUNCA

Additions to rust and chytrid pathogens of Turkey

CuMALI OzasLAN, MAKBULE ERDOGDU, ELsAD HUSEYIN, & ZEKIYE SULUDER

Dentipellicula austroafricana sp. nov. supported by morphological

and phylogenetic analyses —‘Jta-JIA CHEN, Lu-Lu SHEN, & YU-CHENG Dal

A new species of Terriera (Rhytismatales, Ascomycota) on

Photinia villosa Quine Li, YuaN Wu, Dan-Dan Lu, YA-Fer Xu, & YING-REN LIN

New records of one Amygdalaria and three Porpidia species

(Lecideaceae) from China Lu-Lu ZHANG, XIN ZHAO & Line Hu

Anungitea guangxiensis and Ellisembia longchiensis, two new species

from southern China JI-WEN X1A, YING-Ru1 Ma, & X1u-GUo ZHANG

Fistulina subhepatica sp. nov. from China inferred from

morphological and sequence analyses

JE SonG, MEI-LING HAN, & BAo-Kat CuI

Antherospora sukhomlyniae, a new species of smut fungi on

Hyacinthella in Crimea (Ukraine) KyryLo G. SAVCHENKO

Tuber xanthomonosporum, a new Paradoxa-like species from China

YUAN QING, SHU-HONG LI, CHENG-YI Liu, LIN Li, MEI YANG,

XIAO-LEI ZHANG, XIAO-LIN LI, LIN-YONG ZHENG, & YUN WANG

Distance1D - a protein profile analytical program designed for

fungal taxonomy

DuSAN MATERIC, BILJANA KUKAVICA, & JELENA VUKOJEVIC

Coccomyces prominens sp. nov. (Rhytismataceae) on Rhododendron

coeloneurum in China Ya-FeI Xu, YUAN Wu, YAN-QIONG MENG,

SHI-JUAN WANG, & YING-REN LIN

Type studies on Amauroderma species described by J.D. Zhao et al.

and the phylogeny of species in China MENG-Jrz Li & Hal-SHENG YUAN

JANUARY-MARCH 2015... V

Cladonia corymbescens consists of two species

TEUVO AHTI, RAQUEL PINO-Bopas, & SOILISTENROOS 91

Morphology and phylogeny of four new Lactarius species

from Himalayan India

KANAD Das, ANNEMIEKE VERBEKEN, & JORINDE NUYTINCK 105

Synonymy of two species of Bipolaris from aquatic crops of Poaceae

ZI-LAN XIAO, KEVIN D. HYDE, & JING-ZE ZHANG 131

Myxomycetes of Sonora (Mexico) 6. Central plains of the Sonoran Desert

Marcos LIZARRAGA, GABRIEL MORENO, MARTIN ESQUEDA,

CYNTHIA SALAZAR-MARQUEZ, & MARTHA L. CORONADO 145

New taxa of Ambomucor (Mucorales, Mucoromycotina)

from China X1A0-YONG Liu & RU-YONG ZHENG 165

Perenniporia koreana, a new wood-rotting basidiomycete

from South Korea YEONGSEON JANG, SEOKYOON JANG,

YOUNG WOON LIM, CHANGMU KIM, & JAE-JIN Kim 173

New records of corticolous myxomycetes from Turkey

R. BATUR ORAN & C. CEM ErGut 181

A new species of Lophodermium with variously branched paraphyses

Hat-Lin Gu, YA-FEI Xu, DAN-DAN LU, SHI-JUAN WANG, & YING-REN LIN 191

Bertia hainanensis sp. nov. (Coronophorales) from southern China

LARISSA N. VASILYEVA, HaI-X1A Ma,

ALEKSEY V. CHERNYSHEV, & STEVEN L. STEPHENSON 197

Distribution of Alternaria species among sections. 1.

Section Porri PHILIpp B. GANNIBAL 207

Morchella galilaea, an autumn morel from Turkey

HATIRA TASKIN, HASAN HUtsryvIn DoGAN, & SAADET BUYUKALACA 215

Hyphoderma hallenbergii, a new corticioid species from India

MANINDER Kaur, AVNEET P. SINGH, & G.S. DHINGRA 223

Xylaria thailandica- a new species from southern Thailand

NATTHAPACH SRIHANANT, VASUN PETCHARAT, & LARISSA N. VASILYEVA 227

Rosellinia brunneola sp. nov. and R. beccariana new to China

WEI LI & Lin Guo 233

Phyllachora hainanensis sp. nov. from China

Na Liu, LE Wana, Gu Huang, & LIN Guo 237

Two new species of Zasmidium from Nepal RAVINDRA NATH KHARWAR,

ARCHANA SINGH, RAGHVENDRA SINGH, & SHAMBHU KuMAR 241

The lichen genus Leiorreuma in China

X1A0-Hua WANG, LI-LI XU, & ZE-FENG JIA 247

VI ... MYCOTAXON 130(1)

Periconiella liquidambaricola sp. nov. - a new Chinese hyphomycete

Uwe BrAuN, STEFFEN BIEN, Lyp1a HONIG, & BETTINA HEUCHERT 253

First record of Erysiphe syringae-japonicae in Turkey

ILGaz AKATA & VASYL P. HELUTA 259

AFLP characterization of three argentine Coprotus species

ARACELI MARCELA RAMOs, LuIS FRANCO TADIC,

NAHUEL POLICELLI, LAURA INES FERREYRA, & ISABEL ESTHER CINTO 265

Verticicladus hainanensis, a new aquatic hyphomycete

MiNG-TIAN Guo, MIN QIAO, JIAN- YING Li, WEI WANG, & ZE-FEN YU 275

Geographic distribution of Sarcoporia polyspora and

Sarcoporia longitubulata sp. nov.

JOSEF VLASAK, JOSEF VLASAK JR., JUHA KINNUNEN, & VIACHESLAV SPIRIN 279

Coccomyces neolitseae sp. nov. from Zhangjiajie, China

QinG Li, YU-X1A CHEN, CHUN-TAO ZHENG, DAN-DAN LU, & YING-REN LIN 289

Datronia ustulatiligna sp. nov. (Agaricomycetes) from India

HARPREET KAUR, GURPREET Kaur, & G.S. DHINGRA 295

Three new species of Xylaria from China

Gu Huana, RuisHa WANG, LIN Guo, & Na Liu 299

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 130-1 305

ERRATA FROM PREVIOUS VOLUMES

VOLUME 129

p-293, line 2 FOR: HASAN HUSEYIN DOGAN READ: HASAN HUSEYIN DOGAN

p.296, line 32 FOR: Lado, C. READ: Lado C.

p.455, line 28 FOR: DM100 READ: DM1000

p.457, line 3 FOR: Spores ornamented with long ridges, arranged in an incomplete banded

reticulum.

READ: Spores were decorated with long ridges sometimes forming an

incomplete banded reticulum.

p.457, line 17 FOR: (ca. 9-12 um diam.) and unbranched and thicker

READ: (ca. 9-12 um diam.) and thicker

p.458, line 9 FOR: National Basic Research Program of China (3140010180)

READ: National Basic Research Program of China (3140010180) and National

Science and Technology Foundation Project (2014FY210400)

JANUARY-MARCH 2015 ...

REVIEWERS — VOLUME ONE HUNDRED THIRTY (1)

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

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

prepared for this volume.

André Aptroot

N.S. Atri

Z.M. Azbukina

Gerald L. Benny

Lina Bettucci

Uwe Braun

Ana Rosa Burgaz

Lei Cai

Matteo Carbone

Lori Carris

Rafael FE Castaneda- Ruiz

Julia Checa

Gilberto Coelho

Roger T.A. Cook

Bao-Kai Cui

Yu-Cheng Dai

Alejandra Teresa Fazio

Ricardo Galan

Tatiana Baptista Gibertoni

Shouyu Guo

Ian Robert Hall

Nils Hallenberg

Tsutomu Hattori

Vasyl Heluta

Bo Huang

Santosh Joshi

M. Kakishima

Mitko Karadelev

Bryce Kendrick

Daniel P. Lawrence

Hua-jie Liu

Xiao- Yong Liu

Dimuthu S. Manamgoda

Milan Matavulj

Eric H.C. McKenzie

V.A. Melnik

David W. Mitchell

Wieslaw Mulenko

Lorelei L. Norvell

Aysun Peksen

Shaun R. Pennycook

Liliane Petrini

Marcin Piatek

Michele D. Piercey-

Normore

Chaninun Pornsuriya

Adam Rollins

Daniel J. Royse

B.M. Sharma

Roger G. Shivas

Mirjana Stajic

Steven L. Stephenson

Guangyu Sun

Joanne E. Taylor

Michal TomSovsky

Yuri Tykhonenko

Larissa Vasilyeva

Xiang-Hua Wang

Zheng Wang

A.J.S. Whalley

Ming Ye

Alessandra Zambonelli

Xiu-Guo Zhang

Li-Wei Zhou

PUBLICATION DATE FOR VOLUME ONE HUNDRED TWENTY-NINE (2)

MYCOTAXON for OcTOBER-DECEMBER 2014, (I-VI + 215-495)

was issued on January 8, 2015

VII

vill ... MYCOTAXON 130(1)

FOUR STEPS TO SUCCESSFUL MYCOTAXON PUBLICATION IN 2015

Prospective MycotTaxon authors should download instructions PDF, review and

submission forms, and other helpful templates by clicking the ‘file download page’ link

on our INSTRUCTIONS TO AUTHORS page before preparing their manuscript. Below is a

summary of our “4-step’ publication process.

1—PEER REVIEW: Email formatted text and illustration files with a2014 MycoTaxon

Reviewer Comments Form to 2-3 experts for peer review. Authors should (i) ask

peer reviewers to return revisions and comment forms to BOTH authors and Editor-

in-Chief <editor@mycotaxon.com> and (ii) follow reviewer suggestions before

sending revised files to the Nomenclature Editor for nomenclatural review.

2—NOMENCLATURAL REVIEW: Email all text-based files (with phylotrees but No

photos/drawings) to the Nomenclature Editor <PennycookS@LandcareResearch.co.nz>

for accession and pre-submission review. Place “MycoTaxon’ + first author

surname on the subject line; list all peer reviewer names+Email addresses in the

message. The Nomenclature Editor will assign accession numbers and return

annotated files with a list of needed corrections to the authors and Editor-in-Chief.

3—FINAL SUBMISSION: Consult experts, revise and thoroughly proof manuscripts,

and prepare error-free text and image files ready for immediate publication.

Then send the (i) completed 2015 Mycotaxon submission form; (ii) separate

text files for main text, tables, and legends; and (iii) art files to the Editor-in-Chief

<editor@mycotaxon.com>. The Editor-in-Chief usually Emails all coauthors and

expert reviewers within two weeks of final submission, but please wait at least

14 days before sending a follow-up query (without attachments); this helps us

keep Email traffic to a minimum during Mycotaxon publication deadlines or

temporary closures of the editorial office.

4—FINAL EDITORIAL REVIEW & PRESS PREPARATION: Files not ready for publication

will be rejected or returned to authors for further revision; the Editor-in-Chief

gives tentatvely approved manuscripts a final grammatical and scientific review

before converting all files into publishable format. The PDF proof, bibliographic

citation, and nomenclatural entries are sent to all coauthors for final inspection

prior to publication. Thereafter, the Editor-in-Chief corrects ONLY processing or

editorial errors prior to publication but will list corrections of author errors in the

ERRATA of a subsequent volume for no charge. Authors are expected to arrange

payment of page charges and optional open access fees with the Business Manager

<subscriptions@mycotaxon.com> at this time.

MyYcOTAXON LTD— www.mycotaxon.com

The Mycoraxon Webmaster <mycotaxon@gmail.com> posts general and

subscription information, important announcements, and author forms and templates

on the official MycoTaxon site. The server also hosts the regional mycobiota webpage

for free download of distributional annotated species lists.

MYCOTAXON ONLINE— www.ingentaconnect.com/content/mtax/mt

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

subscription articles are offered.

JANUARY-MARCH 2015 ... IX

FROM THE EDITOR-IN-CHIEF

WELCOME TO THE YEAR OF MyYCOTAXON 130! — For several decades we have

determined MycoTaxon volume size by the number of pages (~512/volume). In

recent years, however, our condensed format has — for most papers — produced

pages containing far more words than those found in our older volumes, so that it has

taken longer to fill a volume.

Beginning with Mycoraxon 130, we move to the publication of one volume

(issued quarterly) per year. Pagination will run consecutively through all four issues,

but we will continue to list expert reviewers, errata, and nomenclatural novelties

in each quarterly for immediate reference. Although the subscription period now

coincides with the calendar year, those subscribing to our journal only for 2015 will

still be able to access MycoTAxoONn 130 papers in future years for no extra access fee.

Our new ‘nomenclatural’ change brings us in line with other scholarly journals, most

of which also assess annual subscription rates per ‘volume:

We are pleased to announce that with this move, we are FINALLY back on schedule.

The 2015 JANUARY-MARCH quarterly (this issue) presents all papers submitted to

the Editor-in-Chief as of March 31. No longer tied to a targeted number of pages per

volume, we will be able to deliver future quarterlies promptly and on time. Librarians

(and authors) rejoice!

MyYcoTAXON 130(1) contains 37 research papers by 132 authors (representing 19

countries) and revised by 63 expert reviewers.

Within its pages are 32 species and one variety new to science representing

Ambomucor, Anungitea, Bertia, Coccomyces, Ellisembia, Fistulina, Leiorreuma,

Lophodermium, Periconiella, Phyllachora, Rosellinia, Terriera, Tuber, Verticicladus,

and Xylaria from China; Datronia, Hyphoderma, and Lactarius from India;

Dentipellicula from South Africa; Perenniporia from Korea; Pseudoplectania from

Slovakia and the Czech Republic; Sarcoporia from the Canary Islands and the U.S.A.;

Xylaria (again) from Thailand, and Zasmidium from Nepal. We also offer one new

name in Periconiella, one new Cladonia combination (resulting from the taxonomic

clarification of C. corymbescens), and phylogenetic and pathogenicity confirmation of

the synonymy of Bipolaris zizaniae with B. oryzae.

In addition to range extensions for previously named taxa of myxomycetes (for

Turkey and Mexico) and chytrids, mildews, morels, and rusts (for Turkey), we present

a type study of Amauroderma species from China, a key to Chinese Leiorreuma

species, and a complete list of Alternaria species assigned to A. sect. Porri. The

applicability of Distance 1D protein profile analysis for fungi and AFLP analyses in

Coprotus are also evaluated.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

13 April 2015

MY COTAXON

http://dx.doi.org/10.5248/130.1

Volume 130, pp. 1-10 January-March 2015

Pseudoplectania lignicola sp. nov.

described from central Europe

S. GLEJDURA™, V. KUCERA?, P. LIZON?, & V. KUNCA!?

'Faculty of Ecology and Environmental Sciences, Technical University in Zvolen,

T. G. Masaryka 24, Zvolen 960 53, Slovakia

?Institute of Botany, Slovak Academy of Sciences,

Dubravska cesta 9, Bratislava 845 23, Slovakia

* CORRESPONDENCE TO: glejdura@gmail.com

ABSTRACT — A new species from Slovakia and the Czech Republic, Pseudoplectania lignicola,

is described and illustrated. It is distinguished from other members of the genus by a centrally

arranged globose membranous sheath surrounding the spores, thick ectal excipulum of

oblong cells at the apothecial base, and growth on less specific biotopes. Comparisons with

similar species and the diagnostic significance of membranous sheath surrounding the

ascospores are also discussed.

Keyworps — biodiversity, Sarcosomataceae, typification

Introduction

The genus Pseudoplectania was established by Fuckel (1870) for two fungi,

Peziza nigrella Pers. and Peziza fulgens Pers. Pseudoplectania nigrella was

selected by Seaver (1913) as the lectotype of the genus, and the species itself

was neotypified by Carbone & Agnello (2012). Peziza fulgens is today treated

in the genus Caloscypha Boud.

Eleven species are currently accepted in the genus: Pseudoplectania affinis,

P. carranzae, P. ericae, RP. kumaonensis, P. melaena, P. nigrella, P. ryvardenii,

P. sphagnophila, P. stygia, PR tasmanica, and P. vogesiaca (Carbone et al. 2013,

2014; Iturriaga et al. 2012).

The first finding of the new Pseudoplectania reported here was associated

with Leucobryum glaucum and tentatively identified in situ as P. sphagnophila.

The new fungus was later collected on Abies alba wood buried in soil as well

as among Sphagnum bogs. The species was mostly collected on conifer wood.

2 ... Glejdura & al.

Materials & methods

The study is based on the morphological examination of 11 collections from three

localities in Slovakia and one in the Czech Republic. The macro-morphological characters

were observed in fresh material. The micro-morphological structures were studied in

fresh and dried material using a light microscope with oil immersion lens (x1000).

Vertical sections of material were examined in 5% KOH, tap-water, Melzer’s reagent,

and a Congo Red + ammonia solution. Values of micro-morphological characters

were estimated as average plus and minus a standard deviation of 30 measurements

for each taxon (with 10 and 90 percentiles presented in parentheses). All descriptions

are based on studied specimens. Localities are georeferenced and the coordinates are

in WGS 84 system. Examined specimens are kept in herbarium BRA, SAV, and HR.

The abbreviations of herbaria are cited in accordance with Index Herbariorum (Thiers

2014).

Taxonomy

Pseudoplectania lignicola Glejdura, V. Kucera, Lizon, Kunca, sp. nov. FIGs 1, 2

MycoBank MB 805097

Differs from Pseudoplectania nigrella by ascospores surrounded by a centrally arranged

globose membranous sheath and by elongated cells in the outer ectal excipulum layer

forming a palisade perpendicular to the surface.

Type: Slovakia, Nizke Tatry Mts., ca. 3 km N of Hiadefl village, 48°50’7.38”N

19°18’39.84’E, alt. 700 m, SW exposure, 20° slope, dystric cambisol, biotite granodiorite,

in a 100-year old managed forest: Picea abies (L.) H. Karst. 77%, Abies alba Mill., 17%,

Larix decidua Mill. 6%; on wood of Abies alba, 21.IV.2010, leg. S. Glejdura (Holotype,

BRA CR-19347).

ErymMo.oey: lignicola (Latin) = living on wood. Most specimens were collected from

wood.

APOTHECIA at first hemispherical, later cup shaped, plane at full maturity,

1-2.7 cm diam, substipitate or sessile. Hymenium smooth, dark brown to

blackish brown. Margin composed of thick-walled cylindrical or slightly

clavate hairs 15-110 um long, 5.5-6.1 um broad, with 1-3 septa and 1.5-2.3 um

thick walls. Receptacle densely tomentose, covered with thick-walled flexuose,

coiled, distantly septate (60-185 um between septa), black-brown to black

hairs, <600 um x 4.6-5.5 um, walls 0.9-1.5 um thick.

ECTAL EXCIPULUM 180-200 um thick, outermost zone 2-4 cells thick, cells

dark brown, thick-walled, subglobose to elongated 20-27 x 14-18 um, inner

layer of subhyaline elongated cells forming a palisade perpendicularly oriented

to the surface, cells 34-92 x 15-29 um.

MEDULLARY EXCIPULUM embedded in a gelatinous matrix, outer layer a

180-200 um thick hyaline textura intricata, cells cylindrical, allantoid, narrowly

pyriform 40-60 x 12-22 um. Inner layer a 500-600 um thick hyaline densely

packed textura intricata, hyphae 9-13 um diam. SUBHYMENIUM 30-40 um

Pseudoplectania lignicola sp. nov. (Slovakia) ... 3

FiGurRE 1. Pseudoplectania lignicola (Holotype BRA CR-19347): A. Ectal excipulum, outermost

thick-walled cells. B. Inner layer of the ectal excipulum, elongated cells perpendicularly arranged

to the surface. C. Distantly arranged cells of medullary excipulum. Scale bar = 50 um.

thick, composed of light brown densely arranged cylindric, globose and

ellipsoid cellular hyphae 3-4.6 um diam.

ASCI narrowly cylindrical, tapered below to thin flexuous aporhynchous

base (simple-septate), apex J-, operculate, 8-spored, 259-299 x 12.3-13.7 um.

4 ... Glejdura & al.

o 8

Cc oS

FiGuRE 2. Pseudoplectania lignicola (Holotype BRA CR-19347). A. Ascus with ascospores from

living apothecium; cytoplasmic granules concentrated symmetrically in a limited space by

membranaceous sheath (arrowed). B. Ascospores with centrally arranged membranaceous sheath,

from living apothecium. Scale bar = 10 um.

ASCOSPORES (mature) globose, thick-walled, with granulate or small guttulate

non-oleaginous content (negative reaction in SUDAN 4), (10.2-)11-12.2(-12.3)

Pseudoplectania lignicola sp. nov. (Slovakia) ... 5

um. Mature ascospores surrounded by a membranous cover (sheath) 15.5-19.5

um (including the sheath). The spores are visible in the center of the sheath in

almost all mature ascospores, and in exsiccatae sheath remnants are at least

visible on several spores. Ascospores within membranous sheaths are separated

by cytoplasmic granules laid crosswise in the ascus. Spore walls usually swollen

in 5% KOH, with the membranous sheaths dissolving in the same medium.

PARAPHYSES cylindric, septate, brownish in the upper part and often

agglutinated with an amorphous substance, sometimes branched and

anastomosed in basal and medial parts, 2.2-2.8 um diam, apex mostly straight,

often lobed or diverticulate, with lateral proliferations in upper part, enlarged

to 2.8-3.6 um diam. Irregularly cylindrical paraphyses can be observed in

young or senescent apothecia. Light brown aseptate paraphyses with slightly

thicker walls (so-called hymenial hairs) also present in all stages of apothecial

development.

ECOLOGY & DISTRIBUTION: The species is known from Slovakia and the

Czech Republic and possibly also from Norway (Eckblad 1968, see comment

below). All available collections come from sub-montane and montane moist

coniferous forests (dominated by Picea abies) on acid soils.

ADDITIONAL SPECIMENS EXAMINED: SLOVAKIA, NizkeE Tatry Mrs., ca. 3 km N of

Hiade! village, buffer-zone of Nizke Tatry National Park, 48°50’7.38”N 19°18’39.84’E,

alt. 700 m, in the forest, SW exposition, 20° slope, sporadically rocky, dystric cambisol,

biotite granodiorite, two-storeyed 100-year old production forest, 1“ storey: Picea abies

77%, Abies alba 17%, Larix decidua 6%; 2‘ storey: Fagus sylvatica L. 100%, among moss

Leucobryum glaucum Schimp., 20.I1V.2008, leg. P. Stefanovie (SAV F-10755); on buried

wood of Picea abies, 23.III.2010, leg. S. Glejdura (SAV F-10756); on wood of P. abies,

29.V.2011, leg. S. Glejdura (SAV F-10757); VEPORSKE VRCHY MTs., ca. 1 km SW of

Sihla village, Nature Reserve Vrchslatina, 48°39’18.96”N 19°37’58.50’E, alt. 900 m, in

the forest, flat, 0° slope, dystric histosol (organic soil), diluvial organic sediments (upland

moor) on biotite tonalites and granodiorites, three-storeyed ca 65-year old protective

forest, 1“ storey: Picea abies 100%, 24, 3rd storey: Picea abies 80-90 %, Alnus incana Mill.

20-10%, Sphagno palustris-Piceetum, well-preserved wetland to peatbog biocenoses

with occurrence of specific ecotype of spruce (resonance wood), on buried wood of P.

abies, 28.V.2010, leg. S. Glejdura & V. Kunca (SAV F-10758); on buried wood of P. abies,

leg. S. Glejdura & V. Kunca (SAV F-10759); 2.V1.2010 (SAV F-10760); VYsoKE TAaTRY

Mts., Ticha dolina valley, ca. 6 km NNE of Podbanské village, National Nature Reserve

Ticha dolina, 49°12’1.20’N 19°55’33.24’E, alt. 1140 m, in the forest, NW exposition,

35° slope, typical podzol soil, diluvial sediments on porphyric granitoids and granites,

one-storeyed 55-year old protective forest, Picea abies 65%, Sorbus aucuparia L. 35%, on

dead Sphagnum sp. and needles of P. abies, 18.V.2011, leg. S. Glejdura (SAV F-10761); on

buried wood of Picea abies, among Sphagnum sp. bog, 18.V.2011, leg. S. Glejdura (SAV

F-10762); on cone of Picea abies, 18.V.2011, leg. S. Glejdura (SAV F-10763).

CZECH REPUBLIC, Hraprec KrALove, Orlicka tabule, Chvojnicka plogina,

50°10’45”N 15°54’15”E, alt. 260 m, among moss Leucobryum glaucum in mixed forest

of Picea abies and Pinus sylvestris, 1.1V.2011, leg. Z. Egertova & M. Sochor (HR 89756).

6 ... Glejdura & al.

Discussion

Pseudoplectania is a morphologically and genetically well-delimited genus in

the Sarcosomataceae Kobayasi (Carbone et al. 2013, 2014). Identification of taxa

is based on the paraphysis morphology, ascospore and apothecium size, and

substrate preferences. As we have presented in this paper, another character—

the shape of the gelatinous (mucilaginous) sheath of the spore—must also be

considered an important diagnostic character. Its nature distinguishes similar

species: P. nigrella, P. melaena, and P. lignicola. It is easily detected in fresh

material, less in dry herbarium specimens, although in the type specimen of

P. lignicola the gelatinous sheath was still visible as late as three years after

collection. Spores of P. lignicola lie at the centre of a spherical sheath, whereas

in P. nigrella spores lie to one side of the surrounding sheath (Fics 2, 3). The

hemispherical sheath of spores of P. melaena does not surround the spores but

lies laterally adjacent to the spore surface (Fic 4), a phenomenon that has not

been reported before.

Aside from the spore sheath morphology, Pseudoplectania nigrella (Pers.)

Fuckel differs from P. lignicola by the thickness and textura of the ectal

excipulum as well as by habitat and substrate preferences (on soil in coniferous

forests, rarely on wooden substrates). Also, the ectal excipulum at the base of

its apothecium is thinner (80-100 um) and composed of smaller cells (17-25

um diam) having a textura angularis-globulosa. Eckblad (1968: 119, fig. 65d)

illustrated a specimen of P. nigrella from Norway that has centrally positioned

spores in a membranous sheath. We suspect that this could well represent

P lignicola, so that our new species may have a wider distribution than currently

recorded.

Pseudoplectania melaena (Fr.) Sacc., which has bigger (20-60 mm diam)

stipitate apothecia and apically hooked paraphyses, grows on Abies alba

branches or logs buried in wet soil. Pseudoplectania sphagnophila (Pers.) Kreisel,

associated with Sphagnum sp., has smaller apothecia (10-15 mm diam), and its

paraphyses, which may or may not have lateral branches on the upper parts,

are apically hooked, as in Otidea (Kreisel 1962, Bauer 1999). Pseudoplectania

ericae Donadini, associated with Ericaceae in the Mediterranean area, produces

astipitate apothecia <10 mm diam. and paraphyses that are 2-3-branched at the

apex (Donadini 1987).

Pseudoplectania stygia (Berk. & M.A. Curtis) Sacc. differs mainly by its small

(ca 13 mm diam) apothecia and straight cylindrical paraphyses (Carbone 2013),

while P. ryvardenii Iturr. et al. is distinguished from all other Pseudoplectania

species by its very small (<7 mm diam) apothecia and ornamented ascospores.

Finally, PB kumaonensis Sanwal has distinctly agglutinated paraphyses, and

P. carranzae (Calonge & M. Mata) M. Carbone et al. is separated from other

Pseudoplectania lignicola sp. nov. (Slovakia) ... 7

FiGuRE 3. Pseudoplectania nigrella (SAV F-10764). A. Ascus with ascospores from living apothecium;

cytoplasmic granules concentrated asymmetrically in a limited space by membranaceous sheath

(arrowed). B. Ascospores with eccentrically arranged membranaceous sheath, from living

apothecium. Scale bar = 10 um.

members of the genus mainly by the presence of a tomentum or subicular

hyphae and smaller asci (170-200 x 10-14 um; Calonge & Mata 2002).

8 ... Glejdura & al.

QO;

ao ho

FiGuRE 4. Pseudoplectania melaena (SAV F-10516). A. Ascus with ascospores from living

apothecium; cytoplasmic granules concentrated asymmetrically in a limited space by

membranaceous sheath (arrowed). B. Ascospores with laterally arranged membranaceous sheath,

from living apothecium. Scale bar = 10 um.

Based on our comparison of paraphysis tips in herbarium material of

Pseudoplectania lignicola, P. nigrella, and P. sphagnophila with available literature

data, we conclude that paraphysis shape is not helpful for distinguishing

species. We observed that paraphyses are generally cylindrical, straight, and

unbranched in young apothecia, whereas paraphyses in mature and senescent

apothecia are branched, anastomosed, and often curved at the tip.

Pseudoplectania lignicola sp. nov. (Slovakia) ... 9

The nature of the membranous sheath of the spore, however, is unique

for each Pseudoplectania taxon for which this feature has been observed.

The spores of P lignicola lie at the center of this sheath, and its apothecia are

<27 mm diameter. The new species probably has a wide ecological amplitude,

given its growth on Sphagnum sp., wood, Leucobryum glaucum, and bare soil.

Pseudoplectania species usually occur on the same site for years (when the

conditions allow), and we have registered and collected P lignicola annually

from 2009 to 2013.

In Slovakia P lignicola is rare and possibly endangered (due to lodging and

biotope changes) and should be included in the next edition of the Red List

of Slovak fungi. The fungus has probably been overlooked and incorrectly

identified in the past, but its occurrence in suitable habitats is currently not

seriously threatened.

Acknowledgements

We would like to thank Peter Stefanovie for help in researching the type site of

Pseudoplectania lignicola (Hiadef village), Vaclav Kautman for assistance in the field,

Ginter Bauer for providing material of P. sphagnophila, and Gerard Thijsse and the staff

of the herbarium at Leiden for the opportunity to study Persoon’s herbarium specimens.

Matteo Carbone (Genova, Italy) and Ricardo Galan (Madrid, Spain), who read and

commented on the manuscript, and the support of Shaun Pennycook (Auckland, New

Zealand) are acknowledged. This work was supported by the VEGA [02/0088/13,

02/0150/12] for Kucera and Lizon and by the VEGA [1/1190/12, 1/0362/13] for Glejdura

and Kunca.

Literature cited

Bauer G. 1999. Pseudoplectania sphagnophila (Pers.: Fr.) Kreisel (Ascomycota, Pezizales,

Sarcoscyphaceae) erstmals in Bayern nachgewiesen. Mycol. Bavar. 3: 44—49.

Berkeley MJ. 1875. Notices of North American fungi. Grevillea 3: 145-160.

Calonge FD, Mata M. 2002. Plectania carranzae sp. nov. (Ascomycota) from Costa Rica. Mycotaxon

81: 237-241.

Carbone M. 2013. A type study of Pseudoplectania stygia (Pezizales). Ascomycete.org 5(1): 33-38.

Carbone M, Agnello C. 2012. Studio e tipificazione di Pseudoplectania nigrella. Study and

typification of Pseudoplectania nigrella. Ascomycete.org 4(4): 79-93.

Carbone M, Agnello C, Alvarado P. 2013. Phylogenetic studies in the family Sarcosomataceae

(Ascomycota, Pezizales). Ascomycete.org 5(1): 1-12.

Carbone M, Agnello C, Alvarado P. 2014. Phylogenetic and morphological studies in the genus

Pseudoplectania (Ascomycota, Pezizales). Ascomycete.org 6(1) : 17-33.

Donadini JC. 1987. Etude des Sarcoscyphaceae ss. Le Gal (1). Sarcosomataceae et Sarcoscyphaceae

ss. Korf. Le genre Pseudoplectania emend. nov. P. ericae sp. nov. (Pezizales). Mycol. Helv. 2:

217-246.

Eckblad F-E. 1968. The genera of operculate discomycetes. A re-evaluation of their taxonomy,

phylogeny and nomenclature. Nytt Mag. Bot. 15. 191 p.

Fuckel L. 1870. Symbolae mycologicae. Beitrage zur Kenntnis der rheinischen Pilze. Jahrb. Nass.

Ver. Naturk. 23-24. 459 p.

10 ... Glejdura & al.

Iturriaga T, Mardones M, Urbina H. 2012. A new species of Pseudoplectania (Sarcosomataceae,

Pezizales) from Venezuela. Kurtziana 37(1): 73-78.

Kreisel H. 1962. Pilze der Moore und Ufer Norddeutschlands. III. Pseudoplectania sphagnophila

(Fr. pro var.) Kreisel nov. comb. Westfal. Pilzbriefe 3(5): 74-78.

Saccardo PA. 1889. Sylloge Discomycetum et Phymatosphaeriacearum. Sylloge Fungorum, vol.

8. Padova. 1143 p.Sanwal BD. 1953. Contributions towards our knowledge of the Indian

discomycetes I. Sydowia 7: 191-199.

Seaver FJ. 1913. The genus Pseudoplectania. Mycologia 5(6): 299-302.

http://dx.doi.org/10.2307/3753586

Thiers B. 2014 [continuously updated]. Index Herbariorum: a global directory of public

herbaria and associated staff. New York Botanical Garden's Virtual Herbarium. Available:

http://sweetgum.nybg.org/ih/ [accessed October 2014]

Van Vooren N, Moyne G, Carbone M, Moingeon J-M. 2013. Pseudoplectania melaena (Pezizales):

taxonomical and nomenclatural note. Ascomycete.org 5(1): 47-52.

MYCOTAXON

http://dx.doi.org/10.5248/130.11

Volume 130, pp. 11-15 January-March 2015

Additions to rust and chytrid pathogens of Turkey

CUMALI OZASLAN”, MAKBULE ERDOGDU’,

ELSAD HUSEYIN? & ZEKIYE SULUDERE?

'Dicle University, Faculty of Agriculture, Department of Plant Protection, Diyarbakir-Turkey

?Ahi Evran University, Faculty of Science and Literature, Department of Biology,

Kursehir-Turkey

°Gazi University, Faculty of Science, Department of Biology, Ankara- Turkey

*CORRESPONDENCE TO: cumali.ozaslan@dicle.edu.tr

Asstract — Uromyces bornmuelleri on Bongardia chrysogonum and Physoderma maculare

on Alisma plantago-aquatica have been recently discovered in Turkey. Morphological data

obtained by light and scanning electron microscopy of identified fungi are presented.

Key worps — new records, Chytridiomycota, Basidiomycota

Introduction

The Pucciniaceae are the largest family within the Uredinales, parasitising

almost all major angiosperm orders, with some primarily herbaceous plant

families such as Asteraceae and Poaceae being most commonly infected (van

der Merwe et al. 2007). Uromyces (Link) Unger was raised to generic rank by

Unger (1832), and is typified by U. appendiculatus on Phaseolus. Seventy-four

species of Uromyces have been registered on 227 species of higher plants in

Turkey (Bahcecioglu & Kabaktepe 2012).

All known representatives of the genus Physoderma Wallr.

(Blastocladiomycetes, Physodermataceae) are obligate parasites of vascular

plants (Olson et al. 1980). The generalized life cycle of Physoderma is composed

of two distinct and separate phases, and the ephemeral epibiotic sporangium

with an endobiotic rhizoidal system is confined to a single host cell (Sparrow

& Johns 1965). Physoderma maculare, the type species of the genus, on Alisma

confirms Clinton's (1902) account of the epibiotic stage being produced from

resting zoospores. The latter on mature host tissue may also give rise to the

endobiotic stage, which bears resting spores. On seedlings, however, resting

zoospores produce only epibiotic sporangia (Sparrow 1964).

12 ... Ozaslan & al.

Fics 1-7. Uromyces bornmuelleri. 1: Deformation of host plant. 2: Telia on leaf, in situ. 3: Teliospores.

4-6: Telia and teliospores (SEM). 7. Teliospores (SEM).

Materials & methods

Plant specimens infected with microfungi were collected from Adiyaman and

Siirt provinces, Turkey and prepared according to established herbarium techniques.

Host plants were identified using the Flora of Turkey and East Aegean Islands (Davis

1965-85). The fungal specimens were prepared from the host plants by obtaining thin

sections. Measurements were made from tissues mounted in 5% KOH or tap water.

Microscopical features were examined and microphotographs were made using a Leica

Uromyces and Physoderma spp. new for Turkey... 13

DM E light microscope. Thirty spores were measured for each sample. The microfungi

were identified using relevant literature (De Toni 1888, Saccardo & Sydow 1902,

Gonzalez Fragoso 1918, Kuprevich & Ulijanishchev 1975, van der Merwe et al. 2007).

Species names follow Index Fungorum (2014). The examined specimens have been

deposited in the mycological collection of the Department of Plant Protection, Faculty

of Agriculture, Dicle University, Diyarbakir, Turkey (DUF-M).

For scanning electron microscopy (SEM), 8-10 mm squares of infected leaves were

mounted on SEM stubs with double-sided adhesive tape, coated with gold using a

Polaron SC 502 Sputter Coater, and examined with a Jeol JSM 6060 scanning electron

microscope at 5-10 kV in the Electron Microscopy Unit, Faculty of Science, Gazi

University (Turkey).

Taxonomy

Uromyces bornmuelleri Magnus, Verh. Ges. Deutsch. Naturf. 65: 151.1893. Fics 1-7

SPERMOGONIA, AECIDIA and UREDINIA unknown. TELIA amphigenous,

chestnut brown, in dense groups or scattered, 0.2-3 mm diam., causing leaf

deformation, at first covered by the epidermis, later becoming erumpent,

pulverulent. TELrospores yellow or yellowish-brown, ovoid, globoid,

sometimes oblong, 21-30 x 18-21.5 um, rounded at the apex, rounded or

sometimes attenuate at the base; wall 2-4 um thick, pedicels hyaline, short,

fragile.

SPECIMEN EXAMINED — TURKEY, ADIYAMAN PROVINCE, Center, Kahta, Gerger, Sincik,

Celikhan, Samsat, in wheat field ecosystem, on the leaves of Bongardia chrysogonum (L.)

Spach (Berberidaceae), 12.V.2009, C. Ozaslan CO2029 (DUF-M).

Uromyces bornmuelleri has been reported from Azerbaijan on Leontice

chrysogonum |= Bongardia chrysogonum] (Tranzschel 1939), and from Iraq and

Cyprus on Bongardia chrysogonum (Georghiou 1957, Mathur 1972). Uromyces

bornmuelleri is reported for the first time from Turkey.

Physoderma maculare Wallr., Fl. Crypt. Germ. 2: 192. 1833. Fries 8-13

RESTING SPORANGIA amphigenous, mostly hypophyllous and on leaf

petioles, concentrated on leaves along the nerves, scattered, covered by the

unbroken epidermis, ellipsoid, ovoid, sometimes punctiformis, 1-2 mm diam.,

chestnut brown. RESTING sporEs chestnut brown, globoid, ovoid to ellipsoid,

content granular with more refractive globules, (23.5-)25.5-32 x 21.5-29 um;

with two distinct walls, the inner wall thin and colorless, the outer wall chestnut

brown, 1-2 um thick, smooth. ZoosPorREs not seen.

SPECIMEN EXAMINED — TURKEY, SurtT PROVINCE, Karaca Village, 38°07 09 N

42°01 59 E, in rice field ecosystem, on the leaves of Alisma plantago-aquatica L.

(Alismataceae), 8.V1.2012, C. Ozaslan CO2035 (DUF-M).

The genus Physoderma and P. maculare are reported for the first time from

Turkey. This fungus has been reported from Canada (Conners 1967), Ireland

14 ... Ozaslan & al.

Ae 1 Avr

We.

xSBB SE find ie 41-668.

Figs 8-13. Physoderma maculare. 8: Resting sporangia on leaf, in situ. 9: Resting spores. 10: Resting

sporangia on leaf (SEM). 11-13: Resting spores (SEM).

(Muskett & Malone 1984, as Cladochytrium alismatis), Poland (Czeczuga et

al. 2007), Siberia (De Toni 1888, as Uredo alismatis), Spain (Gonzalez Fragoso

1918, as U. alismatis), and U.S.A. (Farr et al. 1989).

Acknowledgements

The author thanks Prof. Dr. Z.M. Azbukina (Vladivostok, Russia) and Dr. Yuri

Tykhonenko (Kiev, Ukraine) for critically reading the manuscript and serving as

presubmission reviewers. Especially we are grateful to Dr. Shaun Pennycook (Auckland,

Uromyces and Physoderma spp. new for Turkey... 15

New Zealand) for detailed linguistic help and nomenclatural review of the manuscript.

We would like thank DUBAP (Dicle University Research Projects Coordinator) for

financial support of this Project (DUBAP 12ZF71). We also thank Prof. Dr. Mecit Vural

(Gazi University, Ankara) and Prof. Dr. A. Selcuk Ertekin (Dicle University, Diyarbakir)

for his help with the identification of the hosts.

Literature cited

Bahcecioglu Z, Kabaktepe $. 2012. Checklist of rust fungi in Turkey. [Mycotaxon 119: 494.]

http://www.mycotaxon.com/resources/checklists/Bahcecioglu-v119-checklist.pdf

Clinton GP. 1902. Cladochytrium alismatis. Botanical Gazette 33: 49-61.

Conners IL. 1967. An annotated index of plant diseases in Canada and fungi recorded on plants

in Alaska, Canada and Greenland. Research Branch, Canada Department of Agriculture,

Publication 1251. 381 p.

Czeczuga B, Muszynska E, Godlewska A, Mazalska B. 2007. Aquatic fungi and straminipilous

organisms on decomposing fragments of wetland plants. Mycologia Balcanica 4: 31-44.

Davis PH (ed.). 1965-85. Flora of Turkey and East Aegean Islands. Vols 1-9. Edinburgh University

Press, Edinburgh.

De Toni GB. 1888. Sylloge ustilaginearum et uredinearum. Sylloge Fungorum 7(2): 449-882.

Farr DF, Bills GF, Chamuris GP, Rossman AY. 1989. Fungi on plants and plant products in the

United States. APS Press, St Paul MN. 1252 p.

Georghiou GP, Papadopoulos C. 1957. A second list of Cyprus fungi. Government of Cyprus,

Department of Agriculture.

Gonzalez Fragoso R. 1918. La roya de los vegetales. Enumeracion y distribucion geografica de los

Uredales. Conocidos hasta hoy en la Peninsula Iberica e Islas Baleares. Trabajos del Museo

Nacional de Ciencias Naturales, Serie Botanica 15: 1-267.

Index Fungorum. 2014. http://www.indexfungorum.org/names/names.asp. Accessed 26 May 2014.

Kuprevich VF, Ulijanishchev V. 1975. Key to the rust fungi in SSSR. Minsk, Belarus: Nauka i

Tekhnika.

Mathur RS. 1972. Checklist of Iraqi Uredinales. Iraq Natural History Museum, Publication 29:

1=32;

Muskett A, Malone J. 1984. Catalogue of Irish fungi - V. Mastigomycotina and Zygomycotina.

Proceedings of the Royal Irish Academy, B. 84: 83-102.

Olson LW, Edén UM, Lange L. 1980. The endobiotic thallus of Physoderma maydis, the causal agent

of Physoderma disease of maize. Protoplasma 103: 1-16.

Saccardo PA, Sydow P. 1902. Supplementum universale, pars V. Sylloge Fungorum 16. 1291 p.

Sparrow FK. 1964. Observations on chytridiaceous parasites of phanerogams. XIII. Physoderma

maculare Wallroth. Archiv fir Mikrobiologie 48: 136-149.

Sparrow FK, Johns RM. 1965. Observation on chytridiaceous parasites of phanerogams XVI.

Notes on Physoderma from Scirpeae. Archiv fiir Mikrobiologie 51: 351-364.

Tranzschel VG. 1939. Compendium of rusts of the USSR. Botanicheskii Institut Akademii Nauk

SSSR. Moscow. 426 p.

Unger F. 1832 [“1833”]. Die Exantheme der Pflanzen. Wien. 422 p.

van der Merwe M, Ericson L, Walker J, Thrall PH, Burdon JJ. 2007. Evolutionary relationships

among species of Puccinia and Uromyces (Pucciniaceae, Uredinales) inferred from protein

coding gene phylogenies. Mycological Research 111: 163-175.

MY COTAXON

http://dx.doi.org/10.5248/130.17

Volume 130, pp. 17-25 January-March 2015

Dentipellicula austroafricana sp. nov. supported by

morphological and phylogenetic analyses

J1A-JIA CHEN*, Lu-LU SHEN* & YU-CHENG DalI*

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

*CORRESPONDENCE TO: daiyucheng2013@gmail.com

ABSTRACT — Dentipellicula austroafricana sp. nov. is described and illustrated from South

Africa based on morphological characters and rDNA sequence data. It is characterized by

an annual growth habit, resupinate basidiocarps, dense soft spines, a monomitic hyphal

structure with non-amyloid, non-dextrinoid and acyanophilous generative hyphae, absence

of cystidia, presence of cystidioles, and tiny rough basidiospores (2.4-2.9 x 2-2.2 um). A

molecular study based on the combined ITS (internal transcribed spacer region) and nLSU

(the large nuclear ribosomal RNA subunit) dataset supports the new species in Dentipellicula.

A key to accepted species of Dentipellicula is provided.

Key worps — Basidiomycota, Hericiaceae, hydnoid fungi, Russulales, taxonomy, wood-

inhabiting fungi

Introduction

Dentipellis Donk (Russulales, Basidiomycota), typified by D. fragilis (Pers.)

Donk, was introduced for species characterized by an annual growth habit,

hydnoid basidiocarps, soft spines, a monomitic hyphal structure with clamp

connections and cyanophilous hyphae, and amyloid rough basidiospores

(Dai et al. 2009, Ginns 1986, Zhou & Dai 2013). Zhou & Dai (2013), who

demonstrated that Dentipellis was polyphyletic, segregated Dentipellis leptodon

(Mont.) Maas Geest. and Dentipellis tatwaniana Sheng H. Wu from Dentipellis

based on ITS and nLSU rDNA sequences. They proposed Dentipellicula Y.C.

Dai & L.W. Zhou as a new genus for these two hydnoid fungal species, which

are distinguished from Dentipellis by their lack of cyanophilous hyphae.

Dentipellicula taiwaniana (Sheng H. Wu) Y.C. Dai & L.W. Zhou (the generic

type), and D. leptodon (Mont.) Y.C. Dai & L.W. Zhou currently comprise the

entire genus.

* Jra-JiA CHEN and Lu-Lu SHEN contributed equally to this work and share first-author status.

18 ... Chen, Shen, & Dai

During studies on the hydnoid fungi, an undescribed Dentipellicula species

from South Africa was identified based on morphological characters and

phylogenetic analysis of ITS and nLSU sequences. We provide an illustrated

description and include an identification key to all three species representing

Dentipellicula.

Materials & methods

Morphological studies

Sections were studied microscopically according to Dai et al. (2010) at

magnifications <1000x using a Nikon Eclipse 80i microscope with phase contrast

illumination. Drawings were made with the aid of a drawing tube. Microscopic

features, measurements, and drawings were made from sections stained with Cotton

Blue and Melzer’s reagent. Spores were measured from sections cut from the tubes. To

present spore size variation, the 5% of measurements excluded from each end of the

range are given in parentheses. Basidiospore spine lengths were not included in the

measurements. Abbreviations include IKI = Melzer’s reagent, IKI- = negative in Melzer’s

reagent, KOH = 5% potassium hydroxide, CB = Cotton Blue, CB+ = cyanophilous,

CB- = acyanophilous, L = mean spore length (arithmetic average of all spores),

W = mean spore width (arithmetic average of all spores), Q = the L/W ratio, and n

= number of spores measured from given number of specimens. Special color terms

follow Petersen (1996). The studied specimens are deposited in the herbaria as cited

below; collection abbreviations follow Thiers (2014).

DNA extraction and sequencing

A CTAB rapid plant genome extraction kit (Aidlab Biotechnologies, Beijing) was

used to obtain PCR products from dried specimens, according to the manufacturer's

instructions with some modifications (Chen & Cui 2014). The DNA was amplified with

the primers: ITS5 and ITS4 for ITS (White et al. 1990), and LROR and LR7 for nLSU

(Vilgalys & Hester 1990). The PCR procedure for ITS was as follows: initial denaturation

at 95°C for 3 min, followed by 35 cycles at 94°C for 40 s, 54°C for 45 s and 72°C for 1

min, and a final extension of 72°C for 10 min. The PCR procedure for nLSU was as

follows: initial denaturation at 94°C for 1 min, followed by 35 cycles at 94°C for 30 s,

50°C for 1 min and 72°C for 1.5 min, and a final extension of 72°C for 10 min. The PCR

products were purified and sequenced in Beijing Genomics Institute, China with the

same primers.

Phylogenetic analysis

New sequences, deposited in GenBank (TaBLE 1), were aligned with additional

sequences retrieved from GenBank (TaBLE 1) using BioEdit (Hall 1999) and ClustalX

(Thompson et al. 1997). Bondarzewia podocarpi Y.C. Dai & B.K. Cui and B. sp. were

used as outgroup (Zhou & Dai 2013). Prior to phylogenetic analysis, ambiguous

regions at the start and the end of the alignment were deleted and gaps were manually

adjusted to optimize the alignment. The edited alignment was deposited at TreeBase

(http://purl.org/phylo/treebase; submission ID 15859).

Dentipellicula austroafricana sp. nov. (China) ... 19

TABLE 1. Specimens used in ITS and nLSU sequence analyses.

GENBANK ACCESSION NO.

SPECIES SAMPLE NO. LOCALITY

ITS nLSU

B. podocarpi Dai 9261 China KJ583207 KJ583221

Bondarzewia sp. DAOM F-415 Canada DQ200923 DQ234539

Dentipellicula austroafricana Dai 12580 South Africa KJ855274 KJ855275

D. leptodon GB 011123 Uganda EU118625 EU118625

D. taiwaniana Dai 10867 China JQ349115 JQ349101

Cui 8346 China JQ349114 JQ349100

Dentipellis coniferarum Cui 10063 China JQ349106 JQ349092

Yuan 5623 China JQ349107 JQ349093

D. dissita NH 6280 Canada AF506386 AF506386

D. fragilis Dai 12550 China JQ349110 JQ349096

Dai 9009 China JQ349108 JQ349094

D. microspora Cui 10035 China JQ349112 JQ349098

D. parmastoi Cui 8513 China JQ349113 JQ349099

Hericium abietis NH 6990 Canada AF506456 AF506456

H. alpestre NH 13240 Russia AF506457 AF506457

H. americanum DAOM F-21467 Canada AF506458 AF506458

H. cirrhatum Tubingen F794 Germany AF506385 AF506385

H. coralloides NH 282 Sweden AF506459 AF506459

H. erinaceus NH 12163 Russia AF506460 AF506460

Laxitextum bicolor NH 5166 Sweden AF310102 AF310102

Phylogenetic analysis followed Li & Cui (2013). Maximum parsimony (MP) analysis

was performed in PAUP* version 4.0b10 (Swofford 2002). All characters were equally

weighted and gaps were treated as missing data. Trees were inferred using the heuristic

search option with TBR branch swapping and 1000 random sequence additions. Max-

trees were set to 5000, branches of zero length were collapsed and all parsimonious trees

were saved. Clade robustness was assessed using a bootstrap (BT) analysis with 1000

replicates (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 each maximum parsimonious tree (MPT) generated.

Phylogenetic trees were visualized using Treeview (Page 1996).

MrModeltest 2.3 (Nylander 2004) was used to determine the best-fit evolution

model of the combined dataset for Bayesian inference (BI). BI was calculated with

MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003) with a general time reversible (GTR)

20 ... Chen, Shen, & Dai

model of DNA substitution and an invgamma distribution rate variation across sites.

Four Markov chains were performed for 2 runs from random starting trees for 1 million

generations of the combined ITS and nLSU dataset, and trees were sampled every 100

generations. The burn-in was set to discard the first 25% of the trees. A majority rule

consensus tree of all remaining trees was calculated. Nodes that received BT support

>75% and Bayesian posterior probabilities (BPP) 20.95 were considered as significantly

supported.

To determine if the dataset was significantly conflicted, the partition homogeneity

test option in PAUP 4.0b was used between the loci using 1000 replicates and the

heuristic general search option. This test randomly shuffles phylogenetically informative

sites between two paired loci: if the dataset is compatible, shuffling sites between the

loci should not produce summed tree lengths that are significant greater than those

produced by the observed data (Farris et al. 1995; Huelsenbeck et al. 1996).

Molecular phylogeny

Partition homogeneity test showed no conflicts for the two-gene combined

loci (P 20.01). Therefore, in this study, ITS and nLSU were combined into a

single analysis. The combined ITS and nLSU dataset included sequences from

20 fungal collections representing 17 species. The dataset had an aligned length

of 1720 characters, of which 1246 characters are constant, 131 are variable and

parsimony-uninformative, and 343 are parsimony-informative. MP analysis

yielded 2 equally parsimonious trees (TL = 902, CI = 0. 708, RI = 0.786,

RC = 0.557, HI = 0.292). The best model for the combined ITS and nLSU

sequences dataset estimated and applied in the BI was GTR+I+G. BI resulted

in a similar topology with an average standard deviation of split frequencies

= 0.005368 to MP analysis, and thus only the MP tree was provided. Both BT

values (=>50%) and BPPs (20.95) are shown at the nodes (Fic. 1).

The newly sequenced specimen from South Africa was embedded in the

lineage of Dentipellicula (100% MP and 1.00 BPPs) and was closely related to

D. taiwaniana.

Taxonomy

Dentipellicula austroafricana Jia J. Chen, L.L. Shen & Y.C. Dai, sp. nov. Fic. 2

MycoBank MB 811108

Differs from other Dentipellicula species by its exclusively resupinate basidiocarps,

dense spines, absence of gloeoplerous hyphae and cystidia, presence of cystidioles, and

tiny rough basidiospores.

Type — South Africa, KwaZulu-Natal Province, Durban, on charred wood of Ficus,

1.X.2011, Dai 12580 (holotype, BJFC; isotype, IFP; GenBank KJ855274, KJ855275).

ETyMOLoGy — austroafricana (Lat.): referring to South Africa.

FruiTBopy — Basidiocarps annual, resupinate, inseparable, hard corky upon

drying, up to 6 cm long, 2 cm wide, 3 mm thick at center, without odor or taste.

Dentipellicula austroafricana sp. nov. (China) ... 21

87/100 Hericium americanum DAOM F-21467

97/1.00| Hericium alpestre NH 13240

a EF Hericium erinaceus NH 12163

Hericium abietis NH 6990

100/1.00

Hericium cirrhatum Tibingen F794

60/1.00

Hericium coralloides NH 282

Laxitextum bicolor NH 5166

100/0.98| Dentipellis fragilis Dai 12550

99/1.00! ° Dentipellis fragilis Dai 9009

67). Dentipellis dissita NH 6280

Dentipellis parmastoi Cui 8513

100/1.00 | Dentipellis coniferarum Cui 10063

Dentipellis coniferarum Yuan 5623

100/1.00

Dentipellis microspora Cui 10035

86/0.96| Dentipellicula taiwaniana Dai 10867

100/0.99 Dentipellicula tatwaniana Cui 8346

100/1.00 Dentipellicula austroafricana Dai 12580

Dentipellicula leptodon GB 011123

Bondarzewia sp. DAOM F-41

Bondarzewia podocarpi Dai 9261

Fic. 1. Strict consensus tree illustrating the phylogenetic position of Dentipellicula austroafricana,

generated by maximum parsimony method based on ITS+nLSU sequence data. Branches are

labeled with parsimony bootstrap values 250% and Bayesian posterior probabilities >0.95.

Fresh spines soft, white to cream, when dry fragile, cream to buff, up to 2 mm

long, 8-10 per mm across base. Margin cottony, buff to clay-buff, <1 mm wide.

Subiculum very thin, soft corky, buff to reddish brown, <1 mm thick.

HyYPHAL STRUCTURE — Hyphal system monomitic; generative hyphae with

clamp connections, IKI-, CB-; tissues unchanged in KOH.

22 ... Chen, Shen, & Dai

SUBICULUM — Generative hyphae colorless, thick-walled, frequently

branched, interwoven, 4-6 um in diam. Gloeoplerous hyphae absent.

HYMENOPHORAL TRAMA — Generative hyphae colorless, slightly thick-

walled to thick-walled, moderately branched, more or less subparallel along

the spines, 2-5.5 um in diam. Gloeoplerous hyphae absent.

HyMENIUM — Cystidia absent; fusoid cystidioles present, hyaline, thin-

walled, 16.5-18 x 1-4 um; basidia clavate with four sterigmata and a basal

clamp connection, 18-23 x 3.5-5 um; basidioles similar to basidia in shape, but

smaller than basidia. Basidiospores ellipsoid to broadly ellipsoid, thick-walled,

colorless, minutely rough, strongly amyloid, CB+, 2.4-2.9(-3) x (1.8-)2-2.2

(-2.4) um, L = 2.83 um, W = 2.12 um, Q = 1.29-1.39 (n = 60/1).

OTHER SPECIES EXAMINED — Dentipellicula leptodon: CANADA. ONTARIO PROVINCE,

Algonquin Nat. Res., on Betula, 22.X.1966 (DAOM F-158439).

Dentipellicula taiwaniana: CHINA. HAINAN PROVINCE, Ledong County, Jianfengling

Nat. Res., on fallen angiosperm trunk, 12.V.2009, Dai 10872 (BJFC 5114).

Key to accepted species of Dentipellicula

PS BasidiOSPOFES = Sior UI LONG cs firs ade deko ech Ase bet lle & wen ke ¥ wereld Berets Baa D. leptodon

Ty BasidiospOrese<t5ro7 UY TOME Ae eta acto Wie doles hte A heated tet dha ee aa 4 2

2. Spities! 5—/. per Mins -Cystidia, presents #4 6.0% Fwhctus Warhol Whee! hee D. austroafricana

2. Spines 8-10 per mm; cystidia absent ................. 00... eee eee D. taiwaniana

Discussion

Phylogenetically, Dentipellicula austroafricana is recognized in the

Dentipellicula clade and distant from Dentipellis, Hericium Pers., and

Laxitextum Lentz based on the combined ITS and nLSU sequences (Fie. 1).

Morphologically, D. austroafricana is characterized by an annual growth habit,

resupinate basidiocarps, soft and dense spines, a monomitic hyphal structure

with non-dextrinoid generative hyphae, presence of cystidioles, and tiny rough

basidiospores. Both morphology and rDNA sequence data confirmed that

D. austroafricana is a new species in Dentipellicula.

According to the ITS and nLSU-based phylogeny (Fic. 1), Dentipellicula

austroafricana is closely related to D. taiwaniana, which also has annual

basidiocarps and soft spines but which is distinguished by its effused-reflexed

basidiocarps, more scattered spines (5-7 per mm), narrower (2-3.5 um)

generative hyphae in the subiculum, presence of gloeocystidia, and longer

basidiospores (2.8-3.4 x 2.1-2.4 um; Dai et al. 2009, Wu 2007).

Dentipellicula leptodon also has annual basidiocarps and soft spines but

differs from D. austroafricana in its narrower (2-4 um) generative hyphae

in the subiculum, presence of gloeopleurous hyphae in the subiculum and

gloeocystidia, and larger basidiospores (3.6-4.2 x 2.8-3.3 um; Dai et al. 2009,

SSS

——

Lak

Wee

QIFR

OK ial be

eT"

Go a

— f we

Mm I ng a

24 ... Chen, Shen, & Dai

Ginns 1986). Moreover, the two species are separated in the ITS and nLSU-

based phylogenetic analysis (Fie. 1).

Our phylogenetic tree was overall consistent with that presented by Zhou &

Dai (2013): for now, Dentipellis remains polyphyletic based on ITS and nLSU

rDNA sequences, while the new segregate genus Dentipellicula is monophyletic

(Fic. 1). A fully resolved phylogeny for Dentipellis and its related genera requires

evolutionary information from wider taxa samplings and more conserved gene

markers.

Acknowledgements

We express our gratitude to Drs. Li- Wei Zhou (Institute of Applied Ecology, Chinese

Academy of Sciences, China) and Michal TomSovsky (Mendel University in Brno, Czech

Republic) who reviewed the manuscript. The research was financed by the Fundamental

Research Funds for the Central Universities (No. BLYJ201403).

Literature cited

Chen JJ, Cui BK. 2014. Phlebiporia bubalina gen. et. sp. nov. (Meruliaceae, Polyporales) from

southwest China with a preliminary phylogeny based on rDNA sequences. Mycol. Prog. 13:

563-573. http://dx.doi.org/10.1007/s11557-013-0940-4

Cui BK, Zhao CL, Dai YC. 2011. Melanoderma microcarpum gen. et sp. nov. (Basidiomycota) from

China. Mycotaxon 116: 295-302. http://dx.doi.org/10.5248/116.295

Dai YC, Xiong HX, Wu SH. 2009. Notes on Dentipellis (Russulales, Basidiomycota). Mycosystema

28: 668-671.

Dai YC, Cui BK, Liu XY. 2010. Bondarzewia podocarpi, a new and remarkable polypore from

tropical China. Mycologia 102: 881-886. http://dx.doi.org/10.3852/09-050

Farris JS, Kallersj6 M, Kluge AG, Bult C. 1995. Testing significance of incongruence. Cladistics 10:

315-319. http://dx.doi.org/10.1111/j.1096-0031.1994.tb00181.x

Felsenstein J. 1985. Confidence intervals on phylogenetics: an approach using bootstrap. Evolution

39: 783-791. http://dx.doi.org/10.2307/2408678

Ginns J. 1986. The genus Dentipellis (Hericiaceae). Windahlia 16: 35-45.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucleic Acids Symp. Ser. 41: 95-98.

Huelsenbeck JP, Bull JJ, Cunningham E. 1996. Combining data in phylogenetic analysis. Trends

Ecol. Evol. 11: 152-158. http://dx.doi.org/10.1016/0169-5347(96)10006-9

Li HJ, Cui BK. 2013. Taxonomy and phylogeny of the genus Megasporoporia and its related genera.

Mycologia 105: 368-383. http://dx.doi.org/10.3852/12-114

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University.

Page RDM. 1996. TreeView: application to display phylogenetic trees on personal computers.

Comput. Appl. Biosci. 12: 357-358.

Petersen JH. 1996. The Danish Mycological Societys colour-chart. Foreningen til

Svampekundskabens Fremme, Greve. 6 p.

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Swofford DL. 2002. PAUP*: Phylogenetic analysis using parsimony (*and other methods). Version

4.0b10. Sinauer Associates, Sunderland.

Dentipellicula austroafricana sp. nov. (China) ... 25

Thiers, B. 2014 [continuously updated]. Index Herbariorum: A global directory of public herbaria

and associated staff. New York Botanical Garden's Virtual Herbarium.

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

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTAL_X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Res. 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. J. Bacteriol. 172: 4238-4246.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds).PCR Protocols: a guide to

methods and applications. Academic Press, San Diego.

Wu SH. 2007. Three new species of corticioid fungi from Taiwan. Bot. Stud. 48: 325-330.

Zhou LW, Dai YC. 2013. Taxonomy and phylogeny of wood-inhabiting hydnoid species in

Russulales: two new genera, three new species and two new combinations. Mycologia 105:

636-649. http://dx.doi.org/10.3852/12-011

MY COTAXON

http://dx.doi.org/10.5248/130.27

Volume 130, pp. 27-31 January-March 2015

A new species of Terriera (Rhytismatales, Ascomycota)

on Photinia villosa

QinG Lr, YUAN Wu’, DAN-DAN LU’, YA-FEI Xu’, & YING-REN LIN”

" School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT — A new Terriera species, T: aequabilis on fallen leaves of Photinia villosa from

the Mount Sangingshan National Park, Jiangxi Province, China, is described, illustrated,

and discussed. This taxon is distinguished from T. cladophila by smaller ascomata that are

not associated with conidiomata, black thin zone lines, a well-developed excipulum, and

unbranched paraphyses. The type specimen is deposited in the Reference Collection of Forest

Fungi of Anhui Agricultural University, China (AAUF).

KEY worps — taxonomy, morphology, Rhytismataceae, Rosaceae

Introduction

Eriksson (1970) established the genus Terriera based on T. cladophila

(Lév.) B. Erikss. (© Hysterium cladophilum Lév.), which produces a different

paraphysial type and opening mechanism from genera Lophodermium Cheval.

and Sporomega Corda. Minter (1996) provided a detailed description for

T. cladophila. Johnston (2001), who systematically studied monocotyledon-

inhabiting Lophodermium, divided that heterogeneous genus into five groups,

one of which was transferred to Terriera. IndexFungorum (2014) lists 25

specific and two varietal epithets in Terriera.

Here, Terriera aequabilis on fallen leaves of oriental photinia from Jiangxi

Province, China, is described as a new species.

Materials & methods

Mature ascomata were selected from the collected specimens. Under the dissecting

microscope with a 10—50x magnification, the macroscopic appearance of ascomata and

zone lines was observed. After rehydration of reference materials in water for 15 min.,

vertical transverse 10—15um thick sections were sliced from ascomata using a freezing

microtome and mounted in 0.5% (w/v) cotton blue in water for observing ascomatal

28 ... Li & al.

outlines. Gelatinous sheaths surrounding ascospores and paraphyses were examined

in water or 0.1% (w/v) lactophenol-cotton blue. The color of internal structures and

ascospore contents were observed in water. Squash mounts were prepared in 5% KOH

solution for measurements of asci, ascospores, and paraphyses. Figures of the external

shapes and internal structures of the fruitbodies were drawn using a microscopic

painting tube. The type specimen is deposited in the Reference Collection of Forest

Fungi of Anhui Agricultural University, China (AAUF).

Taxonomy

Terriera aequabilis Qing Li & Y.R. Lin, sp. nov. Fires 1-5

MycoBaAnk MB 808818

Differs from Terriera cladophila by smaller ascomata not associated with conidiomata,

frequent black and thin zone lines, a much thinner covering stroma, unbranched

paraphyses swollen conspicuously at the apex, and ascospores with a visible mucous

sheath.

Type: China, Jiangxi, Mount Sanqingshan National Park, Bingyudong, alt. 1550 m,

on fallen leaves of Photinia villosa (Thunb.) DC. (Rosaceae), 21 August 2012, Y.R. Lin,

S.J. Wang & L. Zhang 2657 (Holotype, AAUF 68765).

ErymMo.oey: aequabilis (Latin = uniform), referring to the single ascospore, which has

an equal diameter throughout its length.

Cotontgs on both sides of leaves, forming distinct subcircular or irregular,

yellow-brown bleached areas each with an obvious edge, 6-10 mm diam.,

which tend to coalesce into larger irregular shapes.

ZONE LINES usually frequent, dark brown or black, thin, entirely or partly

surrounding the paler areas.

CONIDIOMATA not observed.

AscomarTa developing on both sides of leaves, predominantly on the lower

side, scattered over the pale areas, with two ascomata sometimes coalescent. In

surface view, ascomata 320-720 x 250-430 um, elliptical to subcircular, straight

or slightly curved to one side, ends rounded. Whole surface of ascoma black,

except sometimes for a paler region at each end, with a clearly defined edge,

slightly shiny, moderately raising above the substrate surface but somewhat

sunken near the split, opening by a single longitudinal split extending most

of the length of the ascoma. Immature ascomata appearing as a single dark

brown protrusion, more or less rotund in outline, not forming a pale zone

along the future line of opening. In median transverse vertical section,

ascomata subepidermal with epidermal cells becoming filled with fungal tissue

as ascoma develops, 165-190 um deep. COVERING STROMA 15-20 um thick

near the opening, becoming thinner towards the edge and connecting to the

basal stroma, composed mainly of textura angularis with blackish-brown to

dark brown, thick-walled cells 3-5.5 um diam. Lip cells absent. Along the edge

of the ascoma opening, there is a short extension, 10-18um thick, adjacent

Terriera aequabilis sp. nov. (China) ... 29

\\ ASS YTES

SURE F) WISSSEY

ORDO US a St

see OS SoS

Fics 1-5. Terriera aequabilis (holotype, AAUF 68765) on Photinia villosa 1. Habit on a leaf. 2.

Detail of ascomata and a zone line. 3. Ascoma in transverse vertical section. 4. Portion of ascoma

in transverse vertical section. 5. Paraphyses, asci, and ascospores.

30 ... Li&al.

to the top of the covering stroma, which is comprised of strongly carbonized

tissue with no obvious cellular structure. ExcrpuULUM well-developed, 10-15

uum thick, arising from the inner layer of the basal stroma, septate, comprised

of colorless textura porrecta-intricata with hyphae ca 0.8 um diam., sometimes

a reticular structure formed by the interlaced hyphae at the upper part. BASAL

STROMA 6-10 um thick, dark brown, consisting of 2(-3) layers of 3-5 um diam.,

angular, thick-walled cells. Colorless to grayish-brown textura prismatica with

cell 3-8.5 um diam., 20-45 um thick, existing in the triangular area, visible in

sections between the covering and basal stroma at each edge of the ascoma.

SUBHYMENIUM 15-22 um thick, rather flat, consisting of hyaline textura-

angularis and textura intricata. PARAPHYSES 1-1.3 um wide, filiform, sparsely

septate, not branched, gradually or suddenly swollen to 2.8-5 um at the apex,

extending ca 10 um beyond asci, immersed in a ca 0.5 um thick gelatinous

matrix, hyphal bridges no observed at the base. Asc1 maturing sequentially,

cylindric-clavate, 75-105 x 4.5-5.5 um, short-stalked, thin-walled, apex round

or slightly obtuse, J-, 8-spored. Ascosporgs arranged fasciculately, with a

uniform diameter throughout the single ascospore, 55-78 x 0.8-1 um, filiform,

hyaline, aseptate, ends rounded, covered by a 0.3-0.5 um thick gelatinous

sheath.

ADDITIONAL SPECIMENS EXAMINED — On dead leaves of Photinia villosa: CHINA,

J1ANGx1: Mount Sanqingshan National Park, Huaiyushan, alt. ca 600 m, 18 August

2012, S.J. Wang & Y.R. Lin 2632 (AAUF 68740); Sanmucun, alt. ca 850 m, 20 August

2012, F. Zhou & L. Zhang 2645 (AAUF 68753); Yulinfeng, alt. ca 810 m, 21 August 2012,

Y.R. Lin, S.J. Wang & L. Zhang 2704 (AAUF 68812).

ComMMENTsS — Terriera aequabilis is somewhat similar to the type species

T. cladophila in the shape of ascomata, asci, and ascospores. However,

T. cladophila occurs on dead or living twigs and has larger (350-900 x

300-600 um) ascomata that are associated with conidiomata, a much thicker

(<40 um) covering stroma composed of textura globulosa or angularis,

paraphyses with frequently and irregularly branched and irregularly twisted

swollen upper parts, sometimes with hyphal bridges at the base between

adjacent paraphyses, ascospores without mucous sheath, and brown diffuse

zone lines occasionally present (Minter 1996).

Terriera minor (Tehon) P.R. Johnst. [= Lophodermium minus (Tehon) P.R.

Johnst.], the most widely distributed species, differs from the new species in

oblong to oblong-elliptic, paraphyses branching 2-3 times in upper 30-40um,

extend 20-30 um beyond asci and sometimes irregularly swollen at the apices,

longer and wider (100-130 x 6-7 um) asci that are tapering abruptly to small,

rounded apex, wider (1.5-2 um) ascospores slightly tapering towards both

ends and 0-1-septate, and a poorly developed subhymenium (Johnston 1989).

In addition, Tehon’s (1918) observation that a thin, hyaline epithecium was

formed by apically coiled paraphyses differs from Johnston's description.

Terriera aequabilis sp. nov. (China) ... 31

The similar Terriera simplex Y.L. Lin et al. is distinguished from T. aequabilis

by larger (650-1000 x 350-480 um), sometimes triangular ascomata with

conidiomata, a very poorly developed excipulum, wider (1.2-1.5 um)

paraphyses occasionally swollen to 2.5-3 um near the apex, synchronously

maturing asci with truncate to obtuse apices, and ascospores tapering to the

rounded base (Gao et al. 2012).

Acknowledgements

The authors are grateful for the pre-submission comments and suggestions provided

by Dr Z. Wang (Yale University, USA) and Dr M. Ye (Hefei University of Technology,

China) and to F Zhou and L. Zhang for the field investigations. This study was supported

by the National Natural Science Foundation of China (No. 31270065, 31170019).

Literature cited

Eriksson B. 1970. On Ascomycetes on Diapensiales and Ericales in Fennoscandia. I. Discomycetes.

Symbolae Botanicae Upsalienses 19(4): 1-71.

Gao XM, Zheng CT, Lin YR. 2012. Terriera simplex, a new species of Rhytismatales from China.

Mycotaxon 120: 209-213. http://dx.doi.org/10.5248/120.209

IndexFungorum. 2014. [www.indexfungorum.org (viewed online on 1 May 2014)].

Johnston PR. 1989. Lophodermium (Rhytismataceae) on Clusia. Sydowia 41: 170-179.

Johnston PR. 2001. Monograph of the monocotyledon-inhabiting species of Lophodermium.

Mycological Papers 176: 1-239.

Minter DW. 1996. Terriera cladophila. IMI Descriptions of Fungi & Bacteria,. no. 1296.

Tehon LR. 1918. Systematic relationship of Clithris. Botanical Gazette 65: 552-555.

http://dx.doi.org/10.1086/332288

SpeciesFungorum. 2014. [www.speciesfungorum.org (viewed online on 1 May 2014)].

MYCOTAXON

http://dx.doi.org/10.5248/130.33

Volume 130, pp. 33-40 January-March 2015

New records of one Amygdalaria and

three Porpidia taxa (Lecideaceae) from China

Lu-Lu ZHANG, XIN ZHAO, & LING Hu’

Key Laboratory of Plant Stress Research, College of Life Sciences, Shandong Normal University,

Jinan, 250014, P. R. China

* CORRESPONDENCE TO: lichenhuling@gmail.com

AxBstRAcT —Four lichen taxa of Lecideaceae, Amygdalaria consentiens var. consentiens,

Porpidia carlottiana, P. lowiana, and P. tuberculosa, are reported for the first time from China.

Key worps —Asia, Lecideales, lichens

Introduction

The family Lecideaceae Chevall. contains about 23 genera and 547 species,

of which the largest genus is Lecidea Ach., containing about 427 species (Kirk

et al. 2008, Fryday & Hertel 2014). In China, twenty-three species of the other

genera in Lecideaceae have been reported, including two each of Amygdalaria

Norman, Bellemerea Hafellner & Cl. Roux, and Immersaria Rambold &

Pietschm.; one each of Lecidoma Gotth. Schneid. & Hertel, Paraporpidia

Rambold & Pietschm., and Stenhammarella Hertel; and 16 species of Porpidia

Korb. (Hertel 1977, Wei 1991; Aptroot & Seaward 1999; Aptroot 2002; Aptroot

& Sparrius 2003; Obermayer 2004; Guo 2005; Zhang et al. 2010, 2012; Wang et

al. 2012; Ismayi & Abbas 2013; Hu et al. 2014).

Amygdalaria and Porpidia are obviously very closely related. Both have

large halonate ascospores, a high hymenium, Porpidia-type asci, and a dark

pigmented hypothecium and are (with a few exceptions) restricted to lime-free,

silicate rocks. However Amygdalaria can be best distinguished from Porpidia

by the presence of cephalodia, the higher hymenium (over 130 um), the larger

ascospores (generally 20-35 x 10-16 um) with conspicuous, rather compact

epispores, and a tendency toward a brownish or yellowish pink thallus (Inoue

1984, Brodo & Hertel 1987, Gowan 1989, Smith et al. 2009).

34 ... Zhang, Zhao, & Hu

During our research on the lecideoid taxa of Lecideaceae in China, we

have identified four taxa new to the country: Amygdalaria consentiens vat.

consentiens, Porpidia carlottiana, P. lowiana, and P. tuberculosa.

Materials & methods

The studied specimens are preserved in the Lichen Section of Botanical Herbarium,

Shandong Normal University, Jinan, China (SDNU); Herbarium Mycologicum

Academiae Sinicae - Lichenes, Beijing, China (HMAS-L); and the Herbarium of

the Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China

(KUN). The morphological and anatomical characters were examined under a stereo-

microscope (COIC XTL7045B2) and a polarizing microscope (OLyMpus CX41). The

thalli and medullae were tested for identification with K (10% aqueous KOH solution),

C (saturated aqueous NaClO solution), I (a 10% aqueous KI solution), P (a saturated

p-phenylenediamine solution in 95% ethyl alcohol). The lichen substances were

identified using standardized thin layer chromatography techniques (TLC) with solvent

system C (Orange et al. 2010). Photos of these lichens were taken under Olympus

SZX16 and BX61 with DP72.

Taxonomic descriptions

Amygdalaria consentiens (Nyl.) Hertel, Brodo & Mas. Inoue, J. Hattori Bot. Lab. 56:

322 (1984) var. consentiens Pret

MorPHoLocy — THALLUS continuous, cracked-areolate, usually thick,

light orange to brownish gray, often becoming oxidated orange; medulla I-;

prothallus black, thin and conspicuous; cephalodia mostly flush with thallus

surface, almost the same colour as the thallus, but occasionally pinker or gray,

containing Stigonema; soredia absent. APOTHECIA entirely sunken in thallus,

(0.35-)0.45-0.9 mm diam., usually one or two per areole; disc dark brown to

black, margin slightly to moderately rusty orange pruinose. Excipie black-

brown, 50-70 um wide, without crystals; epihymenium brownish, with orange

crystals; hymenium hyaline, 150-200 um tall; subhymenium colorless, 45-60

um tall; hypothecium blackish brown, usually thin, bowl-shaped; paraphyses

strongly anastomosed and branched, almost moniliform at the apical part,

with +swollen apices. Asci clavate, Porpidia-type; ascospores hyaline, simple,

ellipsoid, 28-32(-35) x 14-17(-19) um, halonate. Pycnip1A immersed, conidia

bacilliform, 6-9 x c. 1 um.

CHEMISTRY — Thallus and medulla K-, C-, KC-, P-. No lichen substances

were detected by TLC.

DISTRIBUTION — Amygdalaria consentiens has been reported from Asia,

Europe, and North America (Inoue 1984, Brodo & Hertel 1987, Smith et al.

2009). New to China.

SPECIMEN EXAMINED: CHINA. YUNNAN, Luquan, Mt. Jiaozixueshan, alt. 3800 m, on

rock, 26 Oct. 2008, Z.J. Ren 20108488 (SDNU).

Amygdalaria & Porpidia species new to China... 35

Fic. 1 Amygdalaria consentiens var. consentiens (Ren 20108488, SDNU). A: thallus; B: prothallus;

C: apothecium section; D: epihymenium with crystals; E: amyloid reaction of ascus; F: ascus and

ascospores; G: ascospores; H: paraphyses.

COMMENTS —Amygdalaria consentiens var. consentiens is morphologically

similar to A. consentiens var. japonica and A. continua, but A. consentiens var.

japonica contains stictic and constictic acids, while A. continua has a smooth to

finely rimose thallus which is rarely cracked-areolate.

Porpidia carlottiana Gowan, Bryologist 92: 39 (1989) Fia. 2

MorPHoLocy — THALLUS crustose, finely cracked to subrimose, 0.15-0.25

mm thick, whitish gray; medulla I-; prothallus continuous between thallus

36 ... Zhang, Zhao, & Hu

20 pm 20 pm

Eb ad

Fic. 2 Porpidia carlottiana (Wang 20127123, SDNU). A: thallus; B: apothecium section; C: amyloid

reaction of ascus; D: ascospores; E: exciple and epihymenium without crystals.

patches, black, thin; soredia absent. APOTHECIA scattered or contiguous,

sunken in thallus when young, becoming sessile on a broad base when mature,

0.5-1.1(-1.5) mm diam.; disc black, plane to weakly convex, usually with

heavy whitish pruina; margin bare, distinct, regular to flexuose. ExcrpLe dark

brown to black at margin, brown within, 90-105 um wide, without crystals;

epihymenium yellowish brown or olive brown, without crystals; hymenium

hyaline, 90-105(-130) um tall; subhymenium 20-25 um, hypothecium

blackish brown; paraphyses strongly anastomosed and apically branched. Asci

clavate, Porpidia-type; ascospores hyaline, simple, ellipsoid, 17-20 x 7-10 um,

halonate. Pycnip1a not observed.

CHEMISTRY — ‘Thallus and medulla K-, C-, KC-, P-. 2’-O-methyl-

superphyllinic acid was detected by TLC.

DISTRIBUTION — Porpidia carlottiana has been reported from North

America (Gowan 1989). New to China.

SPECIMEN EXAMINED: CHINA. YUNNAN, Lijiang, Mt. Laojunshan, alt. 3800 m, on rock,

5 Nov. 2009, H.Y. Wang 20127123 (SDNU).

ComMENts — Porpidia carlottiana is closely related to P. rugosa, which also

produces pruinose apothecia and 2’-O-methylsuperphyllinic acid. However,

P. rugosa always has soredia, and the apothecia rarely seen.

Amygdalaria & Porpidia species new to China... 37

Porpidia lowiana Gowan, Bryologist 92: 49 (1989) Fic. 3

MorPHOoLocy — THALLUS crustose, continuous, finely cracked to rimose-

areolate, 0.2-0.5 mm thick, yellowish gray to whitish, sometimes dark gray

or greenish gray; medulla I-; prothallus absent; soredia absent. APOTHECIA

scattered to contiguous, soon becoming broadly sessile, 0.5-1.3 mm diam.;

disc black, non-pruinose to moderately pruinose; margin bare, distinct,

even to weakly crenulate, regular to flexuose. ExcrpLe mostly black and

carbonaceous at exciple margin, hyaline within, 55-90 um wide, with crystals

that are soluble in K and C; epihymenium olive brown, without crystals;

hymenium hyaline, 70-110 um tall; subhymenium 18-26 um, hypothecium

blackish brown; paraphyses strongly anastomosed and apically branched. Asci

clavate, Porpidia-type; ascospores hyaline, simple, ellipsoid or tear-drop-shape,

14-16(-25) x (4.5-)6-8 um, halonate. Pycnip1A not observed.

CHEMISTRY — ‘Thallus and medulla K-, C-, KC-, P-. Confluentic,

2’-O-methylmicrophyllinic, and 2’-O-methylperlatolic acids were detected by

TLE.

DISTRIBUTION — Porpidia lowiana has been reported from Europe and

North America (Gowan 1989, Smith et al. 2009). New to China.

Fic. 3 Porpidia lowiana (Ren 20102097, SDNU). A: thallus; B: apothecium section; C: amyloid

reaction of ascus; D: ascospores; E: exciple with crystals.

38 ... Zhang, Zhao, & Hu

SPECIMENS EXAMINED: CHINA. HELONGJIANG, Tahe, Mt. Mengkeshan, alt. 555 m, on

rock, 14 Aug. 2009, Q. Ren 20102097 (SDNU). INNER Monco ia, Mangui, alt. 900 m,

on rock, 13 Sep. 1977, J.C. Wei, 074581 (HMAS-L); SicHUAN, Dukou, Mt. Dabaoding,

alt. 1950 m, on rock, 21 Jun. 1983, L.S. Wang 83-184 (KUN); YUNNAN, Xinping, Mt.

Mopan, alt. 2000 m, on rock, 20 Dec. 2008, L.S. Wang 08-30003 (KUN); Chuxiong, Mt.

Zixi, alt. 2060 m, on rock, 31 Aug. 2005, L.S. Wang 05-25263 (KUN); Lincang county,

Matai village, alt. 1650 m, on rock, 20 Oct. 2003, L.S. Wang 03-22921, 03-22922 (KUN);

Yimen, alt. 1600 m, on rock, 10 Dec. 1990, L.S. Wang 11707 (KUN).

ComMENts — Porpidia lowiana is closely related to P cinereoatra, from

which it differs mainly in having a thinner, yellower thallus, thicker proper

margin (c. 0.05 mm in P. cinereoatra), and sessile apothecia (mainly sunken in

P. cinereoatra).

Porpidia tuberculosa (Sm.) Hertel & Knoph, Beih. Nova Hedwigia 79: 438 (1984)

Fic. 4

MorPHOLOGY — THALLUS crustose, continuous, subrimose to distinctly

rimose-areolate, 0.2-0.3(-0.5) mm thick, medium gray or sometimes rusty

orange; medulla I+ violet; prothallus usually present when abutting another

lichen, black; soralia present, 0.3-0.7 mm diam., round to irregular, scattered

to crowded, shallowly erose or abraded, usually with slightly raised rim;

soredia farinose to granular, white, pale gray or flecked blue-gray or blackish,

I+ violet. APOTHECIA occasional, scattered, soon becoming sessile, 0.9-1.5

(-1.7) mm diam.; disc black, plane to subconvex, weakly to moderately pruinose;

margin bare, distinct, regular to flexuose. ExcIPLE greenish to brownish black

at exciple margin, dark brown within, 90-140 um wide, without crystals;

epihymenium olive brown, without crystals; hymenium hyaline, 90-105

um tall; subhymenium 35-50 um, hypothecium blackish brown; paraphyses

strongly anastomosed and apically branched. Ascr clavate, Porpidia-type;

ascospores hyaline, simple, ellipsoid, 17.5-22 x 7-8 um, halonate. PycNnip1A

immersed, conidia bacilliform, 7.5-10 x c. 1 um.

CHEMISTRY — ‘Thallus and medulla K-, C-, KC-, P-. Confluentic,

2’-O-methylmicrophyllinic, and 2’-O-methylperlatolic acids were detected by

TES

DISTRIBUTION — Porpidia tuberculosa has been reported from Asia,

Macaronesia, Europe, and North & South America (Hertel 1984, Smith et al.

2009). New to China.

SPECIMENS EXAMINED: CHINA. HusBEI, Shennongjia, Mt. Dashennongijia, alt. 3000 m,

on rock, 27 Jun. 1984, J.C. Wei, 067994, 068005, 074292 (HMAS-L). YUNNAN, Luquan,

Mt. Jiaozixueshan, alt. 3800 m, on rock, 27 Oct. 2008, H.Y. Wang 20081180, 20082302

(SDNU).

ComMENtTs — Porpidia tuberculosa, P. rugosa and P. soredizodes all have

a thallus with soralia. However, P rugosa has a more verrucose thallus,

Amygdalaria & Porpidia species new to China... 39

100 um

Fic. 4 Porpidia tuberculosa (Wei 067994, HMAS-L). A: thallus; B: apothecium section; C: amyloid

reaction of ascus; D: conidia; E: exciple and epihymenium without crystals.

I- medulla, and contains 2’-O-methylsuperphyllinic and glaucophaeic acids.

Porpidia soredizodes has an I- medulla as well as an overall smaller thinner

(<0.2 mm) darker thallus that contains stictic acid.

Acknowledgements

The authors thank Dr. A. Aptroot (ABL Herbarium, Soest, the Netherlands) and

Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing, China) for presubmission reviews. This study

was supported by Program for Scientific Research Innovation Team in Colleges and

Universities of Shandong Province, the National Natural Science Foundation of China

(31170187), Foundation of Key Laboratory, CAS (KLBB-201306), and the Scientific

Research Foundation of Graduate School of Shandong Normal University (BCX1406).

Literature cited

Aptroot A. 2002. Corticolous and saxicolous lichens from Xishuangbanna, southern Yunnan,

China. http://www.nhm.uio.no/botanisk/lav/Yunnan.

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

57-101.

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

AO ... Zhang, Zhao, & Hu

Brodo IM, Hertel H. 1987. The lichen genus Amygdalaria (Porpidiaceae) in North America.

Herzogia 7: 493-521

Fryday AM, Hertel H. 2014. A contribution to the family Lecideaceae s. lat. (Lecanoromycetidae inc.

sed., lichenized Ascomycota) in the southern subpolar region; including eight new species and

some revised generic circumscriptions. Lichenologist 46(3): 389-412.

http://dx.doi.org/10.1017/S0024282913000704

Gowan SP. 1989. The lichen genus Porpidia (Porpidiaceae) in North America. Bryologist 92: 25-59.

Guo SY. 2005. Lichens. 31-82, in: WY Zhuang (ed.). Fungi of northwestern China. Mycotaxon Ltd.,

Ithaca, New York.

Hertel H. 1977. Gesteinsbewohnende Arten der Sammelgattung Lecidea (Lichenes) aus Zentral-,

Ost- und Siidasien. Khumbu Himal, Ergebnisse des Forschungsunternehmens Nepal Himalaya.

6: 145-378.

Hertel H. 1984. Uber saxicole, lecideoide Flechten der Subantarktis. Beihefte zur Nova Hedwigia.

79: 399-499.

Hu L, Zhao X, Sun LY, Zhao ZT, Zhang LL. 2014. Four lecideoid lichens new to China. Mycotaxon

128: 83-91. http://dx.doi.org/10.5248/128

Inoue M. 1984. Japanese crustose lichen genera formerly reported under Lecidea sensu lato. 1.

Amygdalaria Norman. Journal of the Hattori Botanical Laboratory 56: 321-330.

Ismayi G, Abbas A. 2013. Bellemerea alpina — new lichen species record from China. Plant Science

Journal 31(4): 333-335.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th edition. CABI

Bioscience: CAB International. 771 p.

Obermayer W. 2004. Additions to the lichen flora of the Tibetan region. Bibliotheca Lichenologica

88: 479-526.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA (eds). 2009. The

lichens of Great Britain and Ireland. Natural History Museum Publications, in association with

The British Lichen Society.

Wang XY, Zhang LL, Joshi Y, Wang HY, Hur JS. 2012. New species and new records of the

lichen genus Porpidia (Lecideaceae) from western China. Lichenologist 44(5): 619-624.

http://dx.doi.org/10.1017/S0024282912000242

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

Zhang LL, Wang HY, Sun LY, Zhao ZT. 2010. Four lichens of the genus Lecidea from China.

Mycotaxon 112: 445-450. http://dx.doi.org/10.5248/112.445

Zhang LL, Wang LS, Wang HY, Zhao ZT. 2012. Four new records of lecideoid lichens from China.

Mycotaxon 119: 445-451. http://dx.doi.org/10.5248/119.445

MYCOTAXON

http://dx.doi.org/10.5248/130.41

Volume 130, pp. 41-46 January-March 2015

Anungitea guangxiensis and Ellisembia longchiensis,

two new species from southern China

JI-WEN XIA, YING-RuI Ma, & XIU-GUO ZHANG

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

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

ABSTRACT — Two new hyphomycete species, Anungitea guangxiensis and Ellisembia

longchiensis, are described and illustrated from specimens collected on dead branches in

southern China. Anungitea guangxiensis is characterized by conidiogenous cells that are

polyblastic, integrated, terminal, smooth, and denticulate (with cylindrical denticles) and

conidia produced in acropetal chains. Ellisembia longchiensis is characterized by distinct,

single, unbranched, erect conidiophores, monoblastic conidiogenous cells, and distoseptate

conidia that have a small, globose, mucilaginous apical sheath.

Key worps — conidial fungi, taxonomy

Introduction

Anungitea was established by Sutton (1973) as a monotypic genus with

A. fragilis B. Sutton as the type species. The genus is characterized by conidia borne

on conspicuous denticulate loci of the terminal or intercalary conidiogenous

cells and which form acropetal, unbranched chains. A further 19 names have

been proposed in Anungitea (Castafeda 1986; Castafieda & Kendrick 1990,

1991; Castaneda et al. 1997; Crous et al. 1995, 2014; Kirk 1983; Matsushima

1975; Rambelli et al. 2008, 2009, 2010; Sutton & Hodges 1978), but subsequently

A. globosa B. Sutton & Hodges was synonymised with A. fragilis (Kirk 1982;

Matsushima 1987; Castaneda Ruiz et al. 1996), and A. triseptata R.F. Castafieda

& W.B. Kendr. was transferred to Anungitopsis (Castaheda & Kendrick 1991).

The genus Ellisembia was introduced by Subramanian (1992) to accommodate

Sporidesmium-like species with determinate or irregularly percurrently

extending conidiogenous cells that produce distoseptate conidia. Wu & Zhuang

(2005) merged Imicles Shoemaker & Hambl. (Shoemaker & Hambleton 2001)

into Ellisembia and expanded the generic concept to include species that

produce typically lageniform, ovoid, or doliiform percurrently extending

conidiogenous cells.

42 ... Xia, Ma, & Zhang

-—-«.

+. a &

.. >

iv.

Peal fe

. -

gue

ad 6

———-.

& . =

al os

“& BLOM a. ee +

lsat Ye

ae = + ae

Fic. 1. Anungitea guangxiensis (holotype, HSAUP H6351).

A,C. Conidiophores with conidia. B. Apex of conidiophore.

D. Conidia. E,F. Apex of conidiophores with conidia.

Many taxa of hyphomycetes have been already described from China (Wu

& Zhuang 2005, Zhang et al. 2009a,b, 2011, 2012, Ma et al. 2010, 2012a,b,c, Ren

Anungitea & Ellisembia spp. nov. (China) ... 43

et al. 2012). In our studies on hyphomycetes from deciduous stems and rotten

wood in southern China, we collected two hyphomycetes that possess the

morphological characteristics of Anungitea and Ellisembia. Both are proposed

herein as new species.

Anungitea guangxiensis J.W. Xia & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 811157

Differs from Anungitea heterospora by its much larger conidia; and from A. longicatenata

by its longer, cylindrical conidia.

Type: China, Guangxi Province: Mount Dayao, on dead stems of unidentified broad-

leaved tree, 26 Oct 2012, J.W. Xia (Holotype, HSAUP H6351; isotype, HMAS 243458).

EryMo_oey: in reference to the province where the type was found.

Colonies on natural substrate effuse, pale brown to brown, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of septate,

pale brown, smooth, 1-2 um wide hyphae. Conidiophores distinct, single,

unbranched, erect, straight or slightly flexuous, cylindrical, smooth, thick-

walled, pale brown to brown, 6-11-septate, 70-160 x 2.5-4.5 um. Conidiogenous

cells polyblastic, integrated, terminal, sympodial, hyaline to pale brown, 10-18 x

2.5-4.5 um, with 1-6 short denticles. Conidia acrogenous, cylindrical, in

unbranched acropetal chains, dry, subhyaline to hyaline, 1-septate, truncate at

the base and rounded at the apex, 16-22 um long, 3-4 um wide.

ComMENTs - Among the known species, Anungitea guangxiensis bears some

resemblance to A. heterospora P.M. Kirk and A. longicatenata Matsush. in

conidial shape. However, the primary conidia of A. heterospora are smaller

(8-9.5 x 3-4 um, l-septate) than those of A. guangxiensis and occasionally

slightly constricted at the septum (Kirk 1983). The conidia of A. longicatenata

differ by being shorter (11-18 x 2.5-4 um, 0-2-septate), narrowly obovoid to

cylindrical, and tapering at both ends (Matsushima 1975).

Ellisembia longchiensis J.W. Xia & X.G. Zhang, sp. nov. Fic. 2

MycoBank MB 811159

Differs from Ellisembia mucicola by its longer, narrower, obclavate conidia with more

numerous distosepta; and from E. suttonii by its longer, narrower conidia and its

conidiogenous cells lacking percurrent proliferations.

Type: China, Sichuan Province: national forest park of Longchi, on dead stems of

unidentified broad-leaved tree, 17 Apr 2012, J.W. Xia (Holotype, HSAUP H6246-1;

isotype, HMAS 243459).

EryMo_oey: in reference to the type locality.

Colonies on natural substrate effuse, pale brown to brown, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of septate, pale

brown, smooth, 2-5 um wide hyphae. Conidiophores distinct, single, erect,

44 ... Xia, Ma, & Zhang

A : B

20um

14> — 1 eo

aed .

(Ory) Gs

Fic. 2. Ellisembia longchiensis (holotype, HSAUP H6246-1).

A. Conidiophores with conidia. B. Conidia.

straight or slightly flexuous, cylindrical, smooth, thick-walled, pale brown

to brown, 2-4-septate, 55-75 x 3.5-5 um. Conidiogenous cells monoblastic,

integrated, terminal, pale brown to brown, 13.5-15 x 3.5-5 um. Conidial

Anungitea & Ellisembia spp. nov. (China) ... 45

secession schizolytic. Conidia holoblastic, solitary, acrogenous, straight or

slightly curved, obclavate, smooth-walled, pale brown to brown, paler towards

the apex where it is hyaline, 9-12-distoseptate, 55-86 um long, 6-7 um thick

in the broadest part, 3-4.5 um wide at the truncate base, with a small, globose,

mucilaginous apical sheath.

Comments - Ellisembia longchiensis is closely related to E. mucicola W.P. Wu

and E. suttonii W.P. Wu. Ellisembia mucicola differs by its shorter, wider,

ellipsoid to fusiform conidia (42-52 x 2.5-4 um, 8-9-septate); and E. suttonii

differs by its shorter, wider conidia (45-52 x 9-12 um, 10-12-septate) and its

percurrently proliferating conidiogenous cells (Wu & Zhuang 2005).

Acknowledgments

The authors express gratitude to Dr. Eric H.C. McKenzie and Dr. Bryce Kendrick 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

Castaneda Ruiz RE 1986. Fungi cubense. Instituto de Investigaciones Fundamentales en Agricultura

Tropical “Alejandro de Humboldt’, C. Habana. 20 p.

Castaneda Ruiz RE, Kendrick WB. 1990. Conidial fungi from Cuba: II. University of Waterloo

Biology Series 33: 1-61.

Castafieda Ruiz RE, Kendrick WB. 1991. Ninety-nine conidial fungi from Cuba and three from

Canada. University of Waterloo Biology Series 35: 1-132.

Castafieda Ruiz RF, Gené J, Guarro J. 1996. Litter hyphomycetes from La Gomera (Canaries).

Mycotaxon 59: 203-215.

Castaneda Ruiz RF, Kendrick WB, Guarro J. 1997. Notes on conidial fungi. XIV. New hyphomycetes

from Cuba. Mycotaxon 65: 93-105.

Crous PW, Wingfield MJ, Kendrick WB. 1995. Foliicolous dematiaceous hyphomycetes from

Syzygium cordatum. Can. J. Bot. 73: 224-234.

Crous PW, Groenewald JZ, Shivas RG. 2014. Fungal Planet 222 - 10 June 2014. Anungitea

eucalyptorum Crous & R.G. Shivas, sp. nov. Persoonia 32: 198-199.

Kirk PM. 1982. New or interesting microfungi. IV. Dematiaceous hyphomycetes from Devon.

Transactions of the British Mycological Society 78: 55-74.

Kirk PM. 1983. New or interesting microfungi. IX. Dematiaceous hyphomycetes from Esher

Common. Transactions of the British Mycological Society 80: 449-467.

Ma LG, Ma J, Zhang YD, Zhang XG. 2010. A new species of Spadicoides from Yunnan, China.

Mycotaxon 113: 255-258. http://dx.doi.org/10.5248/113.255

Ma LG, Ma J, Zhang YD, Zhang XG. 2012a. Spadicoides camelliae and Diplococcium livistonae, two

new hyphomycetes on dead branches from Fujian Province, China. Mycoscience 53: 25-30.

http://dx.doi.org/10.1007/s10267-011-0138-z

Ma J, Zhang YD, Ma LG, Ren SC, Castafieda Ruiz RE, Zhang XG. 2012b. Three new

species of Solicorynespora from Hainan, China. Mycological Progress 11: 639-645.

http://dx.doi.org/10.1007/s11557-011-0775-9

46 ... Xia, Ma, & Zhang

MaJ, Zhang YD, Ma LG, Castaneda Ruiz RE, Zhang XG. 2012c. Three new species of Sporidesmiella

from southern China. Mycoscience 53: 187-193. http://dx.doi.org/10.1007/s10267-011-0152-1

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Matsushima, Kobe. 209 p.

Matsushima T. 1987. Matsushima Mycological Memoirs 5: 1-100.

Rambelli A, Venturella G, Ciccarone C. 2008. Dematiaceous hyphomycetes from Pantelleria

Mediterranean maquis litter. Flora Mediterranea 18: 441-467.

Rambelli A, Venturella G, Ciccarone C. 2009. More dematiaceous hyphomycetes from Pantelleria

Mediterranean maquis litter. Flora Mediterranea 19: 81-113.

Rambelli A, Tempesta S, Venturella G, Ciccarone C. 2010. Dematiaceous hyphomycetes from

Pantelleria Mediterranean maquis litter. Flora Mediterranea 20: 211-233.

Ren SC, Ma J, Ma LG, Zhang YD, Zhang XG. 2012. Sativumoides and Cladosporiopsis,

two new genera of hyphomycetes from China. Mycological Progress 11: 443-448.

http://dx.doi.org/10.1007/s11557-011-0759-9

Shoemaker RA, Hambleton S. 2001. “Helminthosporium” asterinum, Polydesmus elegans, Imimyces,

and allies. Canadian Journal of Botany 79: 592-599. http://dx.doi.org/10.1139/cjb-79-5-592.

Subramanian CV. 1992. A reassessment of Sporidesmium (hyphomycetes) and some related taxa.

Proceeding of the Indian National Science Academy, B 58: 179-190.

Sutton BC. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycological Papers

132: 1-143.

Sutton BC, Hodges CS. 1978. Eucalyptus microfungi. Chaetendophragmiopsis gen. nov. and other

hyphomycetes. Nova Hedwigia 29: 593-607.

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

Diversity Research Series 15: 1-351.

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

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

Zhang YD, Ma J, Ma LG, Castafieda Ruiz RF, Zhang XG. 2011. New species of Phaeodactylium and

Neosporidesmium from China. Sydowia 63: 125-130.

Zhang YD, Ma J, Ma LG, Zhang XG. 2012. Two new species of Taeniolina from southern China.

Mycological Progress 11: 71-74. http://dx.doi.org/10.1007/s11557-010-0729-7

MY COTAXON

http://dx.doi.org/10.5248/130.47

Volume 130, pp. 47-56 January-March 2015

Fistulina subhepatica sp. nov. from China

inferred from morphological and sequence analyses

Jz Sonc, MEI-LING HAN, & BAo-Kai Cur

Institute of Microbiology, Beijing Forestry University, Beijing 100083, China

Beijing Key Laboratory for Forest Pest Control, Beijing Forestry University, Beijing 100083, China

* CORRESPONDENCE TO: cuibaokai@yahoo.com

ABSTRACT — Fistulina subhepatica sp. nov. is described from Yunnan Province, southwestern

China. The main characters of F subhepatica are its pileate basidiomata with rose to reddish

brown pileal surface and white pore surface, individual and easily separable pores (6-9

per mm), a monomitic hyphal structure with clamped generative hyphae, and ellipsoid

basidiospores (4-6 x 3-4 um). Phylogenetic analysis inferred from the combined ITS

(internal transcribed spacer region) and nLSU (the nuclear large subunit ribosomal RNA)

dataset support F. subhepatica as a distinct new species in Fistulina.

Key worps — Agaricales, Agaricomycetes, Fistulinaceae, phylogeny, rDNA, taxonomy

Introduction

Fistulina Bull. was established by Bulliard (1790: t.464) and typified by

F. buglossoides Bull. [= E hepatica (Schaeff.) With.]. It is a small but

cosmopolitan genus and causes a brown rot of hardwoods. Morphologically,

the genus is characterized by an annual growth habit, reddish to brown pileus

surface, separate but closely packed tubes, a monomitic hyphal system, and

ovoid basidiospores (Gilbertson & Ryvarden 1986, Nunez & Ryvarden 2001,

Ryvarden & Gilbertson 1993).

Previous multi-gene phylogenetic studies showed that Fistulina fell into the

euagarics clade and was closely related to Schizophyllum Fr., Auriculariopsis

Maire, and Porodisculus Murrill (Binder et al. 2005, Bodensteiner et al. 2004).

Taxonomic and phylogenetic studies focusing on brown-rot fungi in China

have been carried out recently, and some new species have been described (Cui

2013, Cui & Dai 2013, Cui & Li 2012, Cui et al. 2011, Li & Cui 2013a, Li et al.

2013, Shen et al. 2014, Zhou & Dai 2012, Zhou & Wei 2012). As a continuation

of these surveys, one undescribed species of Fistulina was discovered based on

48 ... Song, Han, & Cui

morphological characters and phylogenetic analysis of the ITS and nLSU rRNA

gene regions.

TABLE 1. List of species, vouchers, and GenBank accession numbers for sequences

used in phylogenetic analysis. New sequences are indicated by bold font.

GENBANK ACCESSION NO.

SPECIES VOUCHER nLsU ITS

Boletus satanas Lenz Bs 2 AF336242 DQ534567

TDB 1000c DQ533973

Coniophora olivacea (Fr.) P. Karst. 402 AF098376

Favolaschia andina Singer KG 0025 HM246679 HM246678

F. calocera R. Heim PDD 70689 AY572006

PDD 71528 AY572007

SR.KEN. 346 AF261417

E cf. calocera JM 98/372 AF261419

EF, peziziformis (Berk. & M.A. Curtis) Kuntze PDD 67440 AY572008

Favolaschia sp. KG 0027 HM246682 HM246681

Fistulina antarctica REG 550 AY571002

CBS 701.85 AY293181 DQ486702

CIEFAP 115 AY571003

EF. hepatica DSH 93-183 AY293182

REG 593 AY571004 AY571038

EF. pallida CBS 508.63 AY571005 AY571039

E. subhepatica Dai 11714 KJ925057

Dai 12416 KJ925053 KJ925058

Cui 11130 KJ925054 KJ925059

Dai 13216 KJ925055 KJ925060

Dai 13244 (HT) KJ925056 KJ925061

Jaapia argillacea CBS 252.74 GU187581 GU187524

Mycena epipterygia (Scop.) Gray GLM 45969 AY207249

M. galericulata (Scop.) Gray GLM 45970 AY207251

TENN 7495 AF261412

RV 87/14.01 AF042636

M. pura (Pers.) P. Kumm. CBH 371 FN394630 KF913023

IS 10/11/2000 FN394634 FN394611

CBH 367 FN394631 KF913022

CBH 358 FN394629 FN394608

M. rosea Gramberg CBH 097 FN394628 FN394556

TL 12393 FN394641 FN394555

UP 2 FN394632 FN394550

M. sanguinolenta (Alb. & Schwein.) P. Kumm. 430360 AB512401 AB512311

Panellus edulis Y.C. Dai et al. HMJAU 7066 GQ219739 GQ219731

HMJAU 7214 GQ219738 GQ219730

Fistulina subhepatica sp. nov. (China) ... 49

Paxillus involutus (Batsch) Fr. UBC F16285 EU486436

Poromycena manipularis (Berk.) R. Heim JM 98/217 AF261423

Porodisculus pendulus HHB-15376-sp AY293204

DAOM 198417 AF261593

Poromycena sp. JM 98/128 AF261429

Suillus cavipes (Opat.) A.H. Sm. & Thiers TDB-646 AF071535

Schizophyllum amplum (Lév.) Nakasone FCUG 1803 AF141873

S. commune Fr. NBRC 4928 AB733339 AB733163

NBRC 6502 AB733340 AB733164

NBRC 30496 AB733341 AB733165

NBRC 30749 AB733342 AB733166

S. fasciatum Pat. CBS 267.60 AF261589 L43385

S. umbrinum Berk. FL 02.1 AF261590 AF249391

HT = holotype

Materials & methods

Morphological studies

The studied specimens are deposited at the herbarium of the Institute of

Microbiology, Beijing Forestry University (BJFC). Microscopic examinations followed

Dai et al. (2010). Sections were studied at magnifications up to 1000x using a Nikon

E 80i microscope and phase contrast illumination (Nikon, Tokyo, Japan). Drawings

were made with the aid of a drawing tube. Microscopic features, measurements, and

drawings were made from slide preparations stained with cotton blue and Melzer’s

reagent. Basidiospores were measured from sections cut from the tubes. In presenting

the variation in the basidiospore size, 5% of measurements that were excluded from

each end of the range are given in parentheses. In the text, the following abbreviations

were used: IKI = Melzer’s reagent, IKI- = negative in Melzer’s reagent, KOH = 5%

potassium hydroxide, CB = cotton blue, CB- = acyanophilous, CB+ = cyanophilous,

L = mean spore length (arithmetic average of all spores), W = mean spore width

(arithmetic average of all spores), Q = variation in the L/W ratios among the specimens

studied, n = number of spores measured from given number of specimens. Special color

terms follow Petersen (1996).

Molecular procedures and phylogenetic analysis

The Phire Plant Direct PCR Kit (Finnzymes, Vantaa, Finland) was used to obtain PCR

products from dried specimens, according to the manufacturer's instructions. A small

piece of dried fungal specimen was lysed in 30 ul dilution buffer for DNA extraction.

After incubating for 3 min at room temperature, 0.75 ul of the supernatant was used

as template for a 30 ul PCR reaction. The internal transcribed spacer (ITS) regions

were amplified with the primers ITS4 and ITS5 (White et al. 1990), and the nuclear

large subunit (nLSU) ribosomal RNA gene regions with the primers LROR and LR7

(http://www. biology.duke.edu/fungi/mycolab/primers.htm). The PCR procedure for

ITS was as follows: initial denaturation at 95°C for 3 min, followed by 34 cycles at 94°C

for 40 s, 54°C for 45 s, and 72°C for 1 min, and a final extension of 72°C for 10 min. The

50 ... Song, Han, & Cui

PCR procedure for nLSU was as follows: initial denaturation at 94°C for 1 min, followed

by 34 cycles at 94°C for 30 s, 50°C for 1 min and 72°C for 1.5 min, and a final extension

of 72°C for 10 min. The PCR products were purified and directly sequenced in Beijing

Genomics Institute, China, with the same primers. All newly generated sequences were

submitted to GenBank (TABLE 1).

Sequences generated for this study were aligned with additional sequences

downloaded from GenBank (TaBLE 1) using BioEdit (Hall 1999) and ClustalX

(Thompson et al. 1997). Prior to phylogenetic analysis, ambiguous positions at the

start and the end were deleted and gaps were manually adjusted to optimize alignment.

Sequence alignment was deposited at TreeBASE (http://purl.org/phylo/treebase;

submission ID 15918).

Phylogenetic analysis was done as in Li & Cui (2013b). Maximum parsimony

analysis and Bayesian inference (BI) was applied to the combined ITS and nLSU dataset.

Jaapia argillacea Bres. was used as outgroup to root trees (Bodensteiner et al. 2004). The

maximum parsimony tree was constructed in PAUP* version 4.0b10 (Swofford 2002).

All characters were equally weighted and gaps were treated as missing data. Trees were

inferred using the heuristic search option with TBR branch swapping and 1000 random

sequence additions. Max-trees were set to 5000, branches of zero length were collapsed

and all parsimonious trees were saved. Clade robustness was assessed using a bootstrap

(BT) analysis with 1000 replicates (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 each maximum parsimonious tree

(MPT) generated.

MrMODELTEST2.3 (Nylander 2004, Posada & Crandall 1998) was used to

determine the best-fit evolution model for BI. BI was calculated with MrBayes3.1.2

(Ronquist & Huelsenbeck 2003) according to the calculated model. Four Markov chains

were run for 2 runs from random starting trees for 1.5 million generations, and trees

were sampled every 100 generations. The first one-fourth generations were discarded as

burn-in. A majority rule consensus tree of all remaining trees was calculated. Branches

that received bootstrap support for maximum parsimony (MP) 275% and Bayesian

posterior probabilities (BPP) 20.95 were considered as significantly supported.

Results

The combined ITS and nLSU dataset include sequences from 49 fungal

collections. The dataset has an aligned length of 1388 characters including

gaps, of which 812 characters are constant, 135 are parsimony-uninformative,

and 441 are parsimony-informative. Maximum parsimony analysis yielded

9 equally parsimonious trees (Length = 1373, CI = 0.650, RI = 0.867, RC = 0.564,

HI = 0.350), and a strict consensus tree of these trees is shown in Fic. 1. The

best model for the combined ITS and nLSU partition is a GTR+I+G model. BI

produced a topology similar to that from MP analysis with an average standard

deviation of split frequencies = 0.007406.

In the phylogenetic tree (Fic. 1), the unknown Chinese Fistulina collections

grouped in a well-supported lineage (MP = 88%, BPP = 1.00), which had a

Fistulina subhepatica sp. nov. (China) ... 51

Favolaschia calocera SR.KEN. 346

a Soins Favolaschia calocera PDD 70689

97/1.00 deleiay Favolaschia calocera PDD 71528

-/0.98| © Favolaschia cf. calocera JM 98/372

IN Favolaschia sp. KG 0027

78/1.00] Favolaschia andina KG 0025

Favolaschia peziziformis PDD 67440

Poromycena manipularis JM 98/217

59/- ||; Mycena galericulata TENN 7495

Mycena galericulata GLM 45970

Mycena epipterygia GLM 45969

Mycena rosea CBH 097

98/0.97 ae Mycena rosea UP 2

Mycena rosea TL 12393

LK) Mycena pura CBH 371

100/1.00 Mycena pura CBH 367

Mycena pura CBH 358

Mycena pura I§ 10/11/2000

89/1.00 Mycena sanguinolenta 430360

Poromycena sp. JM 98/128

Mycena galericulata RV 87/14.01

100/1.00 Panellus edulis HMJAU 7066

Panellus edulis HMJAU 7214

Fistulina subhepatica Dai 11714

Fistulina subhepatica Dai 13244

100/-

99/1.00

88/1.00) Fistulina subhepatica Dai 13216

87/0.99 100/1.00 Fistulina subhepatica Cut 11130

Fistulina subhepatica Dai 12416 Fi stu lin a

100/1.00 99/199 Fistulina hepatica REG 593

Fistulina hepatica DSH 93-183

Fistulina antarctica REG 550

100/1.00 Fistulina antarctica CBS 701.85

98/0.99 100/1.00 64/- Fistulina antarctica CIEFAP 115

Porodisculus pendulus HHB-15376-sp

Porodisculus pendulus DAOM 198417

93/0.99 L_. Fistulina pallida CBS 508.63

Schizophyllum commune NBRC 30749

90/- Schizophyllum commune NBRC 30496

58/- 89/0.99 pelzepioiiuin commune NBRC 4928

Schizophyllum commune NBRC 6502

Schizophyllum fasciatum CBS 267.60

100/1.00

Schizophyllumumbrinum FL 02.1

79/- © Schizophyllumamplum FCUG 1803

66/0.99 — Boletus satanas Bs 2

91/0.98]L Boletus satanas TDB 1000c

89/1.00 Paxillus involutus UBC F16285

Suillus cavipes TDB-646

Coniophora olivacea 402

Jaapia argillacea CBS 252.74

100/-

Figure 1. The phylogenetic position of Fistulina subhepatica based on ITS+nLSU sequence data.

Topology is from maximum parsimony analysis. Bootstrap values >50% and Bayesian posterior

probabilities 20.95 are indicated at the nodes.

sister relationship with E hepatica [= F buglossoides, the generic type]. The new

species is described on the next page.

52... Song, Hany-& Cui

FiGurRE 2. Fistulina subhepatica (holotype), basidiomata in situ. Scale bar = 2 cm.

Taxonomy

Fistulina subhepatica B.K. Cui & J. Song, sp. nov. FIGs. 2-3

MycoBank MB 809111

Differs from Fistulina hepatica by its thin- to slightly thick-walled tramal generative

hyphae with clamp connections and larger basidiospores.

Type: China, Yunnan Province, Jingdong County, Ailaoshan Nature Reserve, on

dead tree of Lithocarpus (Fagaceae), 12.V1I.2013, Dai 13244 (holotype, BJFC 014732;

GenBank KJ925056, KJ925061).

EryMoLoecy: subhepatica (Lat.): refers to the morphological similarity to Fistulina

hepatica.

FruiTBopy — Basidiomata annual, pileate, fleshy, and readily exuding a

reddish blood-like sap when squeezed or bruised when fresh, leathery when

dry. Pileus sessile to laterally substipitate, dimidiate to reniform or subcircular,

projecting up to 25 cm, 30 cm wide, and 6 cm thick at base. Pileal surface rose

to reddish brown when fresh, becoming fuscous to black upon drying, faintly

radially furrowed when fresh; margin acute, concolorous. Pore surface white

when fresh, turning darker when bruised and becoming cinnamon-buff to

reddish brown when dry; pores 6-9 per mm, consisting of individual, crowed

but easily separable tubes. Context clay-buff to black, leathery when dry, up

to 5 cm thick. Tubes concolorous with pore surface, leathery, up to 1 cm long.

Fistulina subhepatica sp. nov. (China) ... 53

iabaateiae aes

10 pm 10 ym

SSS

10 ym

FiGure 3. Fistulina subhepatica (holotype). a: Basidiospores. b: Basidia and basidioles.

c: Cystidioles. d: Cystidial elements from dissepiment edges. e: Hyphae from trama.

f: Hyphae from context. Scale bars = 10 um.

54 ... Song, Han, & Cui

HYPHAL STRUCTURE — Hyphal system monomitic; generative hyphae with

clamp connections, IKI-, CB-, more or less dissolving in KOH.

CONTEXT — Generative hyphae hyaline, thin-walled, rarely branched,

interwoven, 5-12 um in diam, but with inflated portions up to 18 um,

gloeoplerous hyphae present.

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

rarely branched, parallel along the tubes and densely in sections from dried

specimens, 3.5-8 um in diam. Basidia clavate with four sterigmata and a basal

clamp connection, 15-32 x 5-7 um; basidioles in shape similar to basidia, but

slightly smaller. Cystidial elements present in the dissepimental edges, hyaline,

thin-walled, 75-90 x 5-7 um.

Spores — Basidiospores ellipsoid, hyaline, thick-walled, smooth,

IKI-, CB+, 4-6 x 3-4(-4.5) um, L = 4.82 um, W = 3.37 um, Q = 1.36-1.52

(n = 150/5).

ADDITIONAL SPECIMENS EXAMINED — CHINA. YUNNAN PROVINCE, LINCANG,

Xiaodaohe Forest Farm, on dead tree of Castanopsis (Fagaceae), 10.V1I.2013, Dai 13216

(BJFC 014706; GenBank KJ925055, KJ925060); NANHUA County, Dazhongshan Nature

Reserve, on fallen angiosperm trunk, 15.VII.2013, Cui 11130 (BJFC 015245; GenBank

KJ925054, KJ925059); CHuxIoNG, Zixishan Forest Park, on dead tree of Lithocarpus,

28.VUI.2010, Dai 11714 (BJFC 008827; GenBank KJ925057); 11.VI.2011, Dai 12416

(BJFC 010696; GenBank KJ925053, KJ925058).

OTHER SPECIMEN EXAMINED — Fistulina hepatica. FINLAND. VANTAA, Tammosto

Nature Reserve, on dead tree of Quercus (Fagaceae), 8.1X.2012, Dai 12874 (BJFC

013153).

Discussion

The previous reports of Fistulina hepatica in China (Dai 2012, Dai et al.

2011, Wang et al. 2011, Yuan & Dai 2008) were from identifications based only

on morphological characters. According to the current phylogenetic analysis

and more careful morphological examinations, these Chinese samples are

newly described as F. subhepatica. Macroscopically, FE hepatica is quite similar

to FE. subhepatica in its pinkish brown to more reddish or purplish brown pileal

surface, white pore surface when fresh, and individual pores; microscopically,

however, F. hepatica differs from F. subhepatica by having smaller basidiospores,

simple septate and thin-walled trama generative hyphae (Gilbertson &

Ryvarden 1986, Nunez & Ryvarden 2001, Ryvarden & Gilbertson 1993).

Moreover, F. hepatica forms a distinct lineage separate from F. subhepatica

(Fic. 1). In addition, E hepatica is widely distributed in temperate Europe and

grows mainly on Quercus and Castanea, whereas F. subhepatica is found in

subtropical China, occurring mostly on Castanopsis and Lithocarpus. We did

not find FE hepatica on Quercus in northern China, although the big and old

oak trees are suitable for the fungus.

Fistulina subhepatica sp. nov. (China) ... 55

Fistulina antarctica Speg., closely related to F. subhepatica in the phylogenetic

analysis, differs morphologically from FE subhepatica by its longer basidiospores

(5-7 x 3-4 um, Spegazzini 1887).

Fistulina pallida Berk. & Ravenel resembles F. subhepatica in its brownish

pore surface and individual pores; however, it produces smaller basidiomata and

wood-brown pileus surface (Gilbertson & Ryvarden 1986). Phylogenetically,

E pallida falls outside the “core Fistulina clade” and groups with Porodisculus

pendulus (Fr.) Murrill.

Acknowledgments

The authors are grateful to Prof. Yu-Cheng Dai (BJFC, China) for collecting

specimens and improving the text. We express our gratitude to Drs. Li-Wei Zhou

(China) and Tatiana B. Gibertoni (Brazil), who reviewed the manuscript. The research

was financed by Beijing Higher Education Young Elite Teacher Project (YETP0774) and

the National Natural Science Foundation of China (Project No. 31170018).

Literature cited

Binder M, Hibbett DS, Larsson KH, Larsson E, Langer E, Langer G. 2005. The phylogenetic

distribution of resupinate forms across the major clades of mushroom-forming fungi

(Homobasidiomycetes). Systematics and Biodiversity 2: 113-157.

http://dx.doi.org/10.1017/s1477200005001623

Bodensteiner P, Binder M, Moncalvo JM, Agerer R, Hibbett DS. 2004. Phylogenetic relationships

of cyphelloid Homobasidiomycetes. Molecular Phylogenetics and Evolution 2: 501-515.

http://dx.doi.org/10.1016/j.ympev.2004.06.007

Bulliard JBE. 1790. Herbier de la France 10: t.433-480.

Cui BK. 2013. Antrodia tropica sp. nov. from southern China inferred from morphological

characters and molecular data. Mycological Progress 12: 223-230.

http://dx.doi.org/10.1007/s11557-012-0829-7

Cui BK, Dai YC. 2013. Molecular phylogeny and morphology reveal a new species of

Amyloporia (Basidiomycota) from China. Antonie van Leeuwenhoek 104: 817-827.

http://dx.doi.org/10.1007/s10482-013-9994-1

Cui BK, Li HJ. 2012. A new species of Postia (Basidiomycota) from Northeast China. Mycotaxon

120: 231-237. http://dx.doi.org/10.5248/120.231

Cui BK, Li HJ, Dai YC. 2011. Wood-rotting fungi in eastern China 6. Two new species of

Antrodia (Basidiomycota) from Mt. Huangshan, Anhui Province. Mycotaxon 116: 13-20.

http://dx.doi.org/10.5248/116.13

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Cui BK, Liu XY. 2010. Bondarzewia podocarpi, a new and remarkable polypore from

tropical China. Mycologia 102: 881-886. http://dx.doi.org/10.3852/09-050

Dai YC, Cui BK, Yuan HS, He SH, Wei YL, Qin WM, Zhou LW, Li HJ. 2011. Wood-inhabiting fungi

in southern China 4. Polypores from Hainan Province. Annales Botanici Fennici 48: 219-231.

http://dx.doi.org/10.5735/085.048.0302

Felsenstein J. 1985. Confidence intervals on phylogenetics: an approach using bootstrap. Evolution

39:783-791,

Gilbertson RL, Ryvarden L. 1986. North American polypores 1. Fungiflora, Oslo.

56 ... Song, Han, & Cui

Hall TA. 1999. Bioedit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.

Li HJ, Cui BK. 2013a. Two new Daedalea species (Polyporales, Basidiomycota) from South China.

Mycoscience 54: 62-68. http://dx.doi.org/10.1016/j.myc.2012.07.005

Li HJ, Cui BK. 2013b. Taxonomy and phylogeny of the genus Megasporoporia and its related genera.

Mycologia 105: 368-383. http://dx.doi.org/10.3852/12-114

Li HJ, Han ML, Cui BK. 2013. Two new Fomitopsis species from southern China based

on morphological and molecular characters. Mycological Progress 12: 709-718.

http://dx.doi.org/10.1007/s11557-012-0882-2

Nunez M, Ryvarden L. 2001. East Asia polypores 2. Polyporaceae s. lato. Synopsis Fungorum 14:

341-342.

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology

Centre, Uppsala University.

Petersen JH. 1996. Farvekort. The Danish Mycological Society's colour-chart. Foreningen til

Svampekundskabens Fremme, Greve. 6 p.

Posada D, Crandall KA. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics

14: 817-818. http://dx.doi.org/10.1093/bioinformatics/14.9.817

Ronquist F, Huelsenbeck JP. 2003. MRBAYES 3: bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574.

Ryvarden L, Gilbertson RL. 1993. European polypores 1. Synopsis Fungorum 6: 372-374.

Shen LL, Cui BK, Dai YC. 2014. A new species of Postia (Polyporales, Basidiomycota) from

China based on morphological and molecular evidence. Phytotaxa 162: 147-156.

http://dx.doi.org/10.11646/3529

Spegazzini C. 1887. Fungi Patagonici. Boletin de la Academia Nacional de Ciencias en Cordoba 11:

5-64. http://dx.doi.org/10.5962/bhl.title.4071

Swofford DL. 2002. PAUP*: phylogenetic analysis using parsimony (*and other methods). version

4.0b10. Sinauer Associates, Sunderland, Massachusetts.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal_X windows

interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Wang B, Cui BK, Li HJ, Du P, Jia BS. 2011. Wood-rotting fungi in eastern China 5.

Polypore diversity in Jiangxi Province. Annales Botanici Fennici 48: 237-246.

http://dx.doi.org/10.5735/085.048.0304

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols: a guide to

methods and applications. Academic, San Diego.

Yuan HS, Dai YC. 2008. Polypores from northern and central Yunnan Province, Southwestern

China. Sydowia 60: 147-159.

Zhou LW, Dai YC. 2012. Progress report on the study of wood-decaying fungi in China. Chinese

Science Bulletin 57: 4328-4335. http://dx.doi.org/10.1007/s11434-012-5457-8

Zhou LW, Wei YL. 2012. Changbai wood-rotting fungi 16. A new species of Fomitopsis

(Fomitopsidaceae). Mycological Progress 11: 435-441.

http://dx.doi.org/10.1007/s11557-011-0758-x

MYCOTAXON

http://dx.doi.org/10.5248/130.57

Volume 130, pp. 57-59 January-March 2015

Antherospora sukhomlyniae, a new species of smut fungi on

Hyacinthella in Crimea (Ukraine)

KYRYLO G. SAVCHENKO*

Department of Plant Pathology, Washington State University,

Pullman, WA 99164, United States

* CORRESPONDENCE TO: kyryll.savchenko@wsu.edu

AsBstTRACT — Antherospora sukhomlyniae, a new species of smut fungus parasitic on

Hyacinthella sp. in Crimea (Ukraine), is described and illustrated.

Key worps — anther smut, biodiversity, mycobiota, Ustilaginomycetes

Introduction

During a collecting trip for smut fungi in Crimea (Ukraine), carried out

by the author in 2009, several specimens of smut fungi were collected. Based

on the study of the specimen from anthers of Hyacinthella sp. (Asparagaceae),

I propose here a new smut fungus, Antherospora sukhomlyniae, that was

collected in Karadag reserve in the southeastern part of the Crimean peninsula.

Materials & methods

Sorus and spore characteristics were studied using dried herbarium specimens.

Spores were dispersed in a droplet of lactic acid on a microscope slide, covered with

a cover glass, gently heated to boiling point to rehydrate the spores, cooled, and then

examined using a Carl Zeiss Axiostar light microscope (LM) at 1000x magnification.

For scanning electron microscopy (SEM), spores were attached to specimen holders

by double-sided adhesive tape and coated with gold. The surface structure of spores

was observed at 15 kV and photographed with a scanning electron microscope JEOL

JSM-6700EF.

The holotype specimen is conserved in the Herbarium, Department of Biodiversity

and Biotechnology of Fungi, Institute of Evolution, University of Haifa, Israel (HAI).

Taxonomy

Antherospora sukhomlyniae K.G. Savchenko, sp. nov. FIG. 1

MycoBAnk 808616

58 ... Savchenko

FiGurE 1. Antherospora sukhomlyniae (holotype) in flowers of Hyacinthella sp.: spores. a = LM;

b-d = SEM. Bars: a, b= 10 um, c = 2 um, d= 1 um.

Differs from all other Antherospora species by its host specialization on Hyacinthella and

from A. albucae and A. urgineae by its smaller spores, from A. eucomis by its irregularly

shaped spores, and from A. tourneuxii by its thicker spore walls.

Type: Ukraine, Autonomous Republic of Crimea, Karadag reserve, in flowers of

Hyacinthella sp., 5.V.2009, leg. K.G. Savchenko (Holotype HAI 6521).

EryMo_oey: named in honor of the Ukrainian mycologist Maryna M. Sukhomlyn.

Parasitic on Hyacinthella. Sori in all inner floral organs of deformed and

swollen flowers, producing blackish brown, powdery mass of spores, for a long

time enclosed by the outermost floral envelopes. Infection systemic, with all

flowers of the plant infected. Spores ovoid, ellipsoidal, subglobose to irregular,

(6.5-)7-10 x 7.5-13(-14) um, yellowish brown. Spore wall even, ca. 0.8 um

thick, finely densely punctate-verrucose, with finely wavy spore profile.

Antherospora sukhomlyniae sp. nov. (Ukraine) ... 59

DISTRIBUTION - Known only from the type locality in Crimea, on

Hyacinthella sp.

COMMENTS -— Previous studies have demonstrated that while all species of

Antherospora attack hosts in the Asparagaceae, they display a high level of

specificity, with each species restricted to host plants from a single genus, or in

some cases, from a single species (Bauer et al. 2008, Piatek et al. 2011, 2013).

As no Antherospora species has been reported on Hyacinthella, 1 propose a new

species here.

Antherospora sukhomlyniae belongs to the group of Antherospora species

with sori destroying all inner floral organs of infected flowers (Vanky 2012),

which includes A. albucae (Syd. & P. Syd.) R. Bauer et al., A. eucomis Vanky, A.

tourneuxii (A.A. Fisch. Waldh.) R. Bauer et al., and A. urgineae (Maire) R. Bauer

et al. However, these species can be distinguished from A. sukhomlyniae: A.

albucae by its larger spores (7-14.5 x 9.5-22.5 um; Vanky 2012); A. urgineae by

its larger spores (7-12 x 9.5-17.5 um; Vanky 2012); A. eucomis by its regularly

shaped spores with thinner spore walls (0.5 um; Vanky 2009); and A. tourneuxii

by its thinner spore walls (0.5 um; Vanky 2012).

Acknowledgments

The author thanks Lori M. Carris and Vasyl P. Heluta for peer-reviewing the

manuscript, Shaun Pennycook for useful corrections, and Vitalii Sapsai for help with

the SEM microscopy.

Literature cited

Bauer R, Lutz M, Begerow D, Piatek M, Vanky K, Bacigalova K, Oberwinkler F. 2008. Anther smut

fungi on monocots. Mycological Research 112: 1297-1306.

http://dx.doi.org/10.1016/j.mycres.2008.06.002

Piatek M., Lutz M., Chater A.O. 2013. Cryptic diversity of the Antherospora vaillantii complex on

the Muscari species. IMA Fungus 4: 5-19.

Piatek M., Lutz M., Smith P.A., Chater A.O. 2011. A new species of Antherospora supports the

systematic placement of its host plant. IMA Fungus 2: 135-142.

Vanky K. 2009. Taxonomic studies on Ustilaginomycetes - 29. Mycotaxon 110: 289-324.

http://dx.doi.org/10.5248/110.289

Vanky K. 2012. Smut fungi of the world. APS Press, USA.

MY COTAXON

http://dx.doi.org/10.5248/130.61

Volume 130, pp. 61-68 January-March 2015

Tuber xanthomonosporum,

a new Paradoxa-like species from China

YUAN QING", SHU-HONG LI’, CHENG-YI Liv?, LIN Li’, MEI YANG},

XIAO-LEI ZHANG’, XIAO-LIN L14, LIN- YONG ZHENG?* 4, & YUN WANG®®

' Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, School of Life

Science, Sichuan University, 24 (South part) First Ring Road, Chengdu, Sichuan, China

? Biotechnology & Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences,

9 Xueyun Road, Kunming, Yunnan, China

Sichuan Panzhihua Academia of Agriculture and Forestry,

1719 Panzhihua Road, Panzhihua, Sichuan, China

‘Sichuan Academy of Agricultural Sciences, 20 Jingjusi Road, Chengdu, Sichuan, China

‘Department of Light Chemical Engineering, Xichang College,

Mapingba Road, Xichang, Sichuan, China

°Yunnan Institute for Tropical Crop Research, 99 Xuanwei Road, Jinghong, Yunnan, China

* CORRESPONDENCE TO: “zly6559@126.com "wangynz@yahoo.com

ABSTRACT — A new species, Tuber xanthomonosporum, is described based on specimens

collected under Pinus yunnanensis in Panzhihua, Sichuan Province, China. Tuber

xanthomonosporum invariably has only one spore per ascus. It can be distinguished from

the three other Chinese Paradoxa-like species — T: gigantosporum, T: sinomonosporum, and

T. glabrum — by its whitish to yellow-brownish gleba, two-layered peridium, and spiky

cystidia. Molecular analysis also supports T: xanthomonosporum as a unique species.

KEY worps — taxonomy, Pezizales, Yunnan

Introduction

The genus Paradoxa was erected by Mattirolo in 1935 in the Tuberaceae

to accommodate Paradoxa monospora Mattir., a species which, invariably,

has one-spored asci (Montecchi & Sarasini 2000; Leessoe & Hansen 2007).

For the following 50 years the genus remained monotypic until Wang & Hu

(2008) transferred Tuber gigantosporum into Paradoxa. Wang & Li (1991)

originally described T’ gigantosporum based on a single specimen collected

from Huidong County, Sichuan, China, in 1988. The species was characterized

by its extremely large elliptical ascospores and one-spored asci, a feature that

was eventually recognized as being important when a second T: gigantosporum

62 ... Qing & al.

collection was made in Yongren County, Yunnan, China, in 2002. Since then

three other Paradoxa-like species have been reported: Tuber sp. 1 (Kinoshita

et al. 2011), Paradoxa sinensis (Fan et al. 2012), and Tuber glabrum (Fan et

al. 2014). Recent molecular phylogenetic analysis of truffle species supported

Paradoxa within Tuber (Kinoshita et al. 2011, Fan et al. 2013), and subsequently

Fan et al. (2014) transferred P sinensis into Tuber with the replacement name

T. sinomonosporum.

In 2012 during a survey in Panzhihua in the southwestern corner of Sichuan,

China, we found truffles that invariably had one-spored asci. Here we describe

this Paradoxa-like truffle as a new species of Tuber and briefly review the

molecular relationships of truffle species with single-spored asci.

TABLE 1: Origin of the fungal ITS sequences. (New sequences are indicated

in bold font.)

TAXON

Choiromyces meandriformis

Gymnohydnotrya

australiana

Tuber sp. 1

Tuber sp. 2

T. borchii

T. canaliculatum

T. excavatum

T. foetidum

T. fulgens

T. glabrum

T. indicum

T. latisporum

VOUCHER OR CODE

isolate RH691

isolate GB285

isolate OSC130601

isolate K201

isolate K215

clone Bo6

strain Z43

Unknown

JT12670

JT23942

TE-M

isolate zb3281_215

TE-WD

isolate B-2452

isolate B-2489

isolate y466

isolate zb3386_202

isolate M2435

strain T39

strain T47

strain T49

HKAS 44315

HKAS 42380

GEOGRAPHICAL ORIGIN

USA

Japan

Italy

France

USA

Poland

Germany

Poland

Hungary

Germany

USA

China

GENBANK NO.

HM485330

HM485331

JQ925629

AB553344

AB553356

AF106890

AF250291

AF132505

GQ221455

GQ221456

KC330228

HM152020

KC330227

AJ557543

AJ557544

HM152014

HM152024

HM485358

KFO002731

KF002732

JQ638997

JQ639005

JQ639007

DQ898183

DQ898184

Tuber xanthomonosporum sp. nov. (China) ... 63

T. liui HKAS 48269 China DQ898182

T. macrosporum isolate JT 19458 USA HM485372

isolate $7510 Germany JF926121

T. maculatum clone Macl Italy AF106889

1967 New Zealand EU753269

T. melanosporum strainl015 Israel AF167096

LHB-mic-Tmel-Qc-09-1 Spain GU810153

LHB-mic-Tmel-Qr-09-1 Spain GU810152

T. panzhihuanense strain DXJ260 China JQ978644

strain DXJ276 China JQ978650

T. pseudoexcavatum isolate CJ408 USA HM485381

strain E China JQ638958

strain Q China JQ638982

strain T48 China JQ639006

T. rufum 1798 Italy EF362473

1447 Italy EF362477

T. sinomonosporum BJTCFAN150 China KF002729

T. umbilicatum HKAS48267 China GU979032

HKAS44316 China GU979031

isolate T104 China FJ797879

isolate T117 China FJ797880

T. xanthomonosporum YAAS 13185 China KJ162154

YAAS L3186 China KJ162155

YAAS L3187 China KJ162156

T. zhongdianense HKAS 45388B China DQ898186

wang0299 China DQ898187

Materials & methods

Morphological studies

Specimens of a truffle with asci containing a single spore were collected from

Panzhihua, Sichuan, China, in 2012. Specimens are deposited in the Herbarium of

Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural

Sciences, Kunming, Yunnan, China (YAAS).

The samples were examined at the Biotechnology and Germplasm Resources

Institute, Yunnan Academy of Agricultural Sciences, Kunming, China and Sichuan

Panzhihua Academy of Agriculture and Forestry, Panzhihua, Sichuan, China.

Macroscopic characters were described from fresh specimens and microscopic features

described from dried specimens following the methods of Yang & Zhang (2003).

Photographs were made under a Nikon E400 microscope. For scanning electron

microscopy (SEM), spores were scraped from the dried gleba onto double-sided tape

and mounted directly on a SEM stub. They were then coated with gold-palladium and

examined and photographed with a JEOL, JMS-5600LV SEM.

64 ... Qing & al.

DNA amplification & sequence analysis

DNA was extracted using CTAB (Doyle 1987) modified by adding 200 wL 5M

potassium acetate after 4 x CTAB treatments. The primers ITSIF (Gardes & Bruns

1993) and ITS4 (White et al. 1990) were used to amplify the ITS-rDNA region. PCR

reaction solution and cycling parameters used by Chen & Liu (2007) were adopted.

The amplification products were electrophoresed on a 1% agarose gel and purified with

Sangons purification kit. Sequencing was performed with a BigDye® Terminator v3.1

Cycle Sequencing Kit on an ABI 3730XL automatic sequencer.

In order to determine whether the specimens represented a new species, we compared

our Tuber xanthomonosporum ITS-rDNA sequences with 48 ITS-rDNA sequences

downloaded from NCBI. An ITS-rDNA sequence of Gymnohydnotrya australiana B.C.

Zhang & Minter was also included as outgroup. Sequence alignment and phylogenetic

analysis protocols followed Chen & Liu (2007).

Results

Phylogenetic analysis

A total of 52 ITS sequences were included in a phylogenetic analysis

(TABLE 1, PLATE 2). Of the 712 total characters analyzed, 248 are constant,

51 parsimony-uninformative, and 413 parsimony-informative (L = 1979,

CI = 0.481, RI = 0.812, RC = 0.390). ITS sequence analysis revealed that the

three Paradoxa-like species (Tuber sp. 1, T: sinomonosporum, T. glabrum) form

a well-supported subclade (bootstrap support = 98%) within Clade I, which

also included T: macrosporum, T. canaliculatum, and Tuber sp. 2. In contrast,

T. xanthomonosporum does not cluster with the other Paradoxa-like species

but forms a new Subclade I in Clade II (with low bootstrap support) alongside

Subclade II (/melanosporum) and Subclade III (/rufum).

Taxonomy

Tuber xanthomonosporum Qing & Y. Wang, sp. nov. PLATE 1

MycoBAnk MB 807668

Differs from other Tuber species with single-spored asci by its whitish to yellow-

brownish gleba and two-layered peridium with spiky cystidia.

TyPE: China, Sichuan Province, Panzhihua, 26°29’N 102°01’E, under Pinus yunnanensis

Franch., 15 Nov. 2012: Y. Qing Pan1201 (Holotype, YAAS L3185; GenBank KJ162154).

EryMoLoecy: xanthomonosporum from Latin refers to the yellow brownish gleba and

single spored asci.

PiaTE 1. Tuber xanthomonosporum (YAAS L3185, holotype): 1, 2. Fresh ascomata showing

surface features and cut sections. 3. Cross section of peridium showing two layers of pseudo-

parenchymatous and interwoven tissue with some cystidia. 4. Cross section of outer layer of

peridium showing two kinds of cystidia: talland tapered (arrows), and short and obtuse (arrowhead).

5-7. Ascospores and asci (invariably with only one spore). 8. Ascospores showing ornamentations.

Scale bars: 1, 2 = 1 cm; 3, 8 = 20 um; 4, 7 = 50 um; 5 = 100 um; 6 = 40 um.

66 ... Qing & al.

Ascomata subglobose or irregular and much lobed with deep furrows, firm

and a little rubbery, whitish to yellow-brownish, up to 3.0 cm broad, rough

with hairs. Opor slightly garlicky when mature. PERIDIuM 160-200 um

thick, composed of two layers; the outer layer complex, 50-70 um thick,

pseudoparenchymatous, composed of irregular globose to rectangular cells,

19-32 x 13-15 um diam and thin-walled; the outmost cells giving rise at the

surface to cystidia of two types; one tall, thin, tapered, reaching 50-90 x 2-4 um

and sometimes with a small teat on the wall, hyaline; the other short, obtuse,

1-2-septate, 15-20 x 4-5 um; the inner layer 110-130 um thick, composed of

mainly interwoven, colourless hyphae of 2-4 um diam with some big cells of

2-4 um diam, that gradually merge into the gleba tissue. GLEBA solid, whitish

when young, becoming yellow-brownish at maturity, marbled with distinct,

white and meandering veins, merging at many points with the peridium. Asc1

60 x 70 um, globose to subglobose, sessile, slightly thick walled 2-3 um thick,

invariably 1-spored and randomly dispersed in glebal tissue. AscospoREs

globose, 41-43 x 43-46 um; spore walls 2.5 um thick, brown at maturity,

ornamented with a regular alveolate-reticulum, 2-3 um deep, constituted of

mostly hexagonal meshes 8-15 x (5-)7-13 um, 4-5 across the spore width.

ECOLOGY & DISTRIBUTION — Hypogeous, solitary or in groups in calcareous

soils with pH 6.8-7.6 under Pinus yunnanensis and forming ectomycorrhizae

with its roots, alt. 1500-2000 m, fruiting from autumn to winter. Known only

from Sichuan, China.

ADDITIONAL SPECIMENS EXAMINED: CHINA, SICHUAN PROVINCE, Panzhihua,

26°29’N 102°01’E, under Pinus yunnanensis, 15 Nov. 2012, C.Y. Liu Pan-No-10 (YAAS

L3186; GenBank KJ162155); C.Y. Liu Pan-No-40 (YAAS L3187; GenBank KJ162156).

Discussion

The new Tuber species, T: xanthomonosporum from Sichuan, produces only

one spore per ascus, the diagnostic character of the now defunct genus, Paradoxa

(Mattirolo 1935; Fan et al. 2014). Recent phylogenetic studies of Paradoxa-like

species (Kinoshita et al. 2011; Fan et al. 2014) found that they group with Tuber

species. Our phylogenetic analysis groups T. xanthomonosporum with Tuber

species but not with other Paradoxa-like species, despite sharing the common

character of having only one spore per ascus. Our work therefore supports

the conclusion by Fan et al. (2014) that Paradoxa-like truffle species do not

form a monophyletic group. While Tuber sinomonosporum [= P. sinensis],

T. glabrum, and T. sp. 1 (the Japanese Paradoxa-like new species) do group

together, they also group with T! macrosporum and T. canaliculatum in Clade

I (/macrosporum). These species in Clade I share the same morphological

feature of having red-brown gleba. Tuber xanthomonosporum groups in Clade

II with the /melanosporum and /rufum subclades but with very low bootstrap

support (<50%). Morphologically, T. xanthomonosporum differs greatly from

Tuber xanthomonosporum sp. nov. (China) ... 67

Gymnohydnotrya australiana JQ925629

98 Tuber sp. 1 AB553344

T. sinomonosporum KF002729

100» 7. glabrum KF002731

100 T. glabrum KF002727 Clade I

Tuber sp. 2 AB553356 macrosporum

100) 7. canaliculatum GQ221455

T. canaliculatum GQ221456

100-- 7. macrosporum HM485372

T. macrosporum JF926121

T. xanthomonosporum KJ162154

T. xanthomonosporum KJ162155 Subclade I

T. xanthomonosporum KJ162156

T. indicum JQ638997

T. indicum JQ639005

100 T. indicum JQ639007

T. melanosporum AF167096

100

08 ye T. melanosporum GU810153 Subelade II

T. melanosporum GU810152 melanosporum

100r 7. pseudoexcavatum HM485381 Clade II

100 T. pseudoexcavatum JQ638958

& 100 7. pseudoexcavatum JQ638982

T. pseudoexcavatum JQ639006

T. rufum EF362473

100 T. rufum EF362477

T. rufum JF926123

T. umbilicatum GU979032_ | Subclade TIT

T. umbilicatum GU979031 rufum

go) 2. umbilicatum FJ797879

95! 7. umbilicatum FJ797880

T. borchii AF106890

T. borchii AF250291

83 T. borchii AF132505

100 T. liui DQ898182

T. zhongdianense DQ898186

1001 7, zhongdianense DQ898187

100) 7. latisporum DQ898183

100 T. latisporum DQ898184

109 T. panzhihuanense JQ978644

100! 7. panzhihuanense JQ978650

100) 7. foetidum AJ557543

100 T. foetidum AJ557544

100 7. maculatum AF106889

T. maculatum EU753269

T. excavatum KC330228

loop 7. excavatum HM152020

T. excavatum KC330227

T. fulgens HM152014

100 T. fulgens HM152024

T. fulgens HM485358

100 Choiromyces meandriformis HM485330

Choiromyces meandriformis HM485331

100 100

100

_10

PLATE 2. One of eight most parsimonious trees constructed with ITS sequences of Tuber

xanthomonosporum and related species. MP Bootstrap values greater than 50% are indicated at

nodes.

the species in these two subclades in its whitish to yellow-brownish gleba and

reticulate spores. Given the few species and low support for the species now

placed in Clade I, the phylogenetic relationships need further study with more

samples and gene loci.

Acknowledgements

We are grateful to Dr. Ian Hall and Dr. A. Zambonelli for reviewing the paper. We

also thank Prof. Fan Li and Mr. H.K. Xiong for providing Paradoxa-like specimens for

our study. This study was supported financially by the National Science Foundation

of China (No.31160010) and the Science and technology support program of Sichuan

Province (No.2012NZ0003 and No.2013NZ0029).

68 ... Qing & al.

Literature cited

Chen J, Liu PG. 2007. Tuber latisporum sp. nov. and related taxa, based on morphology and DNA

sequence data. Mycologia 99: 475-481. http://dx.doi.org/10.3852/mycologia.99.3.475

Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure from small quantities of fresh leaf

tissues. Phytochemical Bulletin 19: 11-15.

Fan L, Cao JZ, Li Y. 2012. Tuber microsphaerosporum and Paradoxa sinensis spp. nov. Mycotaxon

120: 471-475. http://dx.doi.org/10.5248/120.471

Fan L, Feng S, Cao JZ. 2014. The phylogenetic position of Tuber glabrum sp. nov. and T. sino-

monosporum nom. nov., two Paradoxa-like truffle species from China. Mycological Progress

13(2): 241-246. http://dx.doi.org/10.1007/s11557-013-0908-4

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes-

application to the identification of mycorrhizae and rusts. Molecular Ecology 2(2): 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Kinoshita A, Sasaki H, Nara K. 2011. Phylogeny and diversity of Japanese truffles (Tuber

spp.) inferred from sequences of four nuclear loci. Mycologia 103(4): 779-794.

http://dx.doi.org/10.3852/10-138

Lzessoe T, Hansen K. 2007. Truffle trouble: what happened to the Tuberales? Mycological Research

111(9): 1075-1099. http://dx.doi.org/10.1016/j.mycres.2007.08.004

Mattirolo O. 1935. Catalogo ragionato dei funghi ipogei raccolti nel Canton Ticino e nelle provincie

Italiane confinanti. Beitrage zur Kryptogamenflora der Schweiz 8: 1-53.

Montecchi A, Sarasini M. 2000. Fungi ipogei d’Europa. A.M.B. Fondazione Centro Studi

Micologici. Vicenza.

Wang Y, Hu HT. 2008. New species of Paradoxa gigantospora comb. nov. from China. Mycotaxon

106: 199-202.

Wang Y, Li ZP. 1991. A new species of Tuber from China. Acta Mycologica Sinica 10(4): 263-265.

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, New York.

Yang ZL, Zhang LE. 2003. Type studies on Clitocybe macrospora and Xerula furfuracea var. bispora.

Mycotaxon 88: 447-454.

MY COTAXON

http://dx.doi.org/10.5248/130.69

Volume 130, pp. 69-72 January-March 2015

Distance1D - a protein profile analytical program

designed for fungal taxonomy

DuSAN MATERIC??’, BILJANA KUKAVICA}, & JELENA VUKOJEVIC4

'Faculty of Agriculture & Teachers Training Faculty, University of East Sarajevo,

Semberskih ratara bb, 76300 Bijeljina, Bosnia and Herzegovina

*Faculty of Science, The Open University, Walton Hall, MK7 6AA Milton Keynes, UK

*Faculty of Natural Sciences and Mathematics, University of Banja Luka,

Mladena Stojanovica 2, 78000 Banja Luka, Bosnia and Herzegovina

‘Faculty of Biology, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia

* CORRESPONDENCE TO: dusan.materic@gmail.com

ABSTRACT —Taxonomic analysis of macromycete fruiting bodies is a challenging task

that utilizes morphological, biochemical, and molecular methods. Many biochemical and

molecular methods have been developed to test or confirm identifications or phylogenetic

positions independently of morphological data. SDS electrophoresis has been shown to be a

good biochemical method for protein separation. Although protein profiles can be analyzed

by commercially available software, there is no software designed specifically for fungal

taxonomic research. We have developed an open source portable program that uses protein

profiles of fungal fruiting bodies to calculate relative differences between species for use in

generating to generate more accurate phylogenetic trees.

Key worps — biochemistry, fungi, Perl, proteome

Introduction

The challenging task of macromycete taxonomy uses morphological,

biochemical, ecological, and molecular data (Guarro et al. 1999, Blackwell et

al. 2006, Korabecna 2007, Lutzoni & Vilgalys 1995). Sodium dodecyl sulfate

(SDS) electrophoresis is a good biochemical method for protein separation

and biomarker discovery, which when applied to the proteomes of fungal

fruiting bodies gives a good number of separated proteins, revealing small (or

sometimes large) differences in protein expression (Materi¢ 2012). The method,

which is robust and reliable, could serve as an independent taxonomic tool for

fungi (Guarro et al. 1999, Tyrrell 1969).

70 ... Materi¢, Kukavica, & Vukojevi¢é

The aim of this work was to develop a user-friendly open source program

using electrophoresis data for fungal taxonomy. Such a program would compare

and calculate relative differences between taxa based on results from SDS gel

electrophoresis and similar techniques (e.g., western blot, zymography).

Materials & methods

We sampled and identified 21 fruiting bodies representing differently related

species. Proteins were separated according to the method described in Materic (2012),

and protein profiles were obtained for each species. The quality of protein profiles

depends upon many factors during sample homogenization, protein extraction and

electrophoresis, and an optimized protocol can be found in Materic¢ et al. (2012). In this

work we have chosen to show the results of only seven species: however, more detailed

dendrograms are available in Materi¢ (2012).

Protein profile data are stored in .csv format and examples of files can be found along

with the source-code in Materi¢ (2013). Protein profile files include two sets of data:

(1) molecular weights (MW) of proteins; and (2) relative abundance. MW and relative

abundances can be obtained using specialized software such as TotalLab (Phoretix,

Newcastle, United Kingdom). However, abundances should be recalculated as relative

abundances with range of 1 (least abundant) to 5 (most abundant). In order to create

a distance matrix table, protein profiles are compared by distancelD. The distance

matrix tables are transformed into dendrograms using the program package PHYLIP

(Felsenstein 2002).

Results & discussion

Protein electrophoresis, which has been widely used in solving taxonomic and

systematic problems, produces results that are independent of morphological

and molecular data (Guarro et al. 1999, Tyrrell 1969). Thus, it is crucial to

have appropriate software that deals with taxonomic issues rather than with

pure biochemical properties. Apart from distance1D, there is no such software

specifically designed to interpret fungal taxonomy from protein profile data.

Distance1D is a program designed to calculate relative differences between

two protein profiles, which could then be used to generate dendrograms

(Fic. 1). The program is designed and written specifically for use in fungal

taxonomy (mainly for analyzing protein profiles of fungal fruiting bodies), but

its usage could be wider. The program is meant to be open source, user-friendly,

and portable. Portability results from the program's being written in Perl, which

can be run on any operating system. As the program uses the Tk library for

generating widgets (Windows-like interface), Perl/Tk should be installed along

with Perl.

Our previous work produced good protein profiles of fungal fruiting bodies

from SDS gel electrophoresis (Materi¢ 2012) that are stored in separate files

(Fie. 1):

Distance 1D for analyzing fungal profiles ... 71

[ SDS Electrophoresis

Fruiting bodies (Boletus edulis)

distancelD 0.1

Image analysis and storing

Species 1: | /GELS2-3-2Be.lin protein profiles in a .csv files

Species 2: /GELS/2-3-3Sg.lin

TOTAL DIF: 150.7

(mwa):

(abd):

Za > 2 ;

pa Boletus edulis

Boletus luridus

Distance matrix yee

Distances obtained using distance1D

Boletusedulis _|000.0[093.9]217.5] 134.6] 170.0]188.1]150.7

Boletus luridus | 093.9]000.0|182.8]151.0/169.6]1903]161.8) Neighbour joining

Lactarius piperatus — |217.5|182.8/000.0/ 113.0] 133.3]131.0]166.1

Suillus granulatus

Russula cyanoxantha

Tree drawing using

Lactarius piperatus

Russula foetens

Russula emetica

FIGURE 1. Diagram showing stages of an experiment used in order to gain biochemically based

phylogenetic trees. A: Fruiting bodies. B: Result of SDS electrophoresis (channels: M = molecular

mass marker; 1 = Boletus luridus; 2 = B. edulis; 3 = Suillus granulatus; 4 = Russula foetens;

5 = Lactarius piperatus; 6 = R. cyanoxantha; 7 = R. emetica; Materi¢ et al 2012). C: Interface of

the program distance1D. D: Distance matrix table generated by the program. E: Phylogenetic tree

drawn by PHYLIP program package (similar results were obtained by analyzing ITS II regions of

rDNA as control; Materi¢ 2012).

The program as an input takes the paths of two files (two file names) where

protein data are stored in .csv format. An algorithm searches for optimal pairing

of proteins by looping through the parameters, such as MW sensitivity. After

alignment and optimization, the following differences are scored: (1) presence

of new protein lines (new protein), (2) molecular weight difference of protein

pairs, and (3) abundance difference of protein pairs. The scoring system,

which could be adapted according to user requirements, is set up as follows:

“10” scored for each new protein line, “1” scored for each 1kDa MW shift, and

“2” scored for abundance difference. If DNA/RNA gel data are going to be used,

the abundance difference for each fragment should be set the same.

There are several important advantages to using this program: the program

generates distances that give accurate phylogenetic trees (more accurate than

a single DNA sequence such as rDNA ITS1; Materi¢ 2012); the parameters

can be changed to suit particular research; the program can be easily adapted

72 ... Materié, Kukavica, & Vukojevi¢

for use with any protein/DNA/RNA electrophoresis data; and the program is

suitable for all operating systems and is open source and free.

Acknowledgments

The authors thank Milan Matavulj (University of Novi Sad, Serbia) and Mirjana

Stajic (University of Belgrade, Serbia) for presubmission review.

Literature cited

Blackwell M, Hibbett DS, Taylor JW, Spatafora JW. 2006. Research Coordination Networks: a

phylogeny for kingdom Fungi (Deep Hypha). Mycologia 98: 829-837.

http://dx.doi.org/10.3852/mycologia.98.6.829

Felsenstein, J. 2002. PHYLIP (Phylogeny Inference Package) version 3.6 a3.

Guarro J, Gené J, Stchigel AM. 1999. Developments in fungal taxonomy. Clinical Microbiology

Reviews 12: 454-500.

Korabecna M. 2007. The variability in the fungal ribosomal DNA (ITS1, ITS2, and 5.8S rRNA

gene): its biological meaning and application in medical mycology. 783-787 in Communicating

current research and educational topics and trends in applied microbiology (A Méndez- Vilas,

ed.). Formatex, Spain.

Lutzoni F, Vilgalys R. 1995. Integration of morphological and molecular data sets in estimating

fungal phylogenies. Canadian Journal of Botany 73: 649-659.

http://dx.doi.org/10.1139/b95-307

Materi¢ D. 2012. Biohemijska, molekularna i bioinformaticka analiza taksona gljiva podcarstva

Dikarya. Master's thesis, Bosnia and Herzegovina, University of Banja Luka.

Materi¢ D. 2013. Distance1D - source code. GitHub. https://github.com/dusanac/distance1D

Materi¢ D, Kukavica B, Boroja M, Vukojevic¢ J. 2012. Optimizacija protokola za ekstrakciju proteina

iz plodonosnih tijela gljiva (vrste rodova: Boletus, Russula, Lactarius i Agaricus) za SDS-

elektroforezu. Skup 4(1):36-41.

Tyrrell D. 1969. Biochemical systematics and fungi. Botanical Review 35: 305-316.

http://dx.doi.org/10.1007/BF02858875

MY COTAXON

http://dx.doi.org/10.5248/130.73

Volume 130, pp. 73-77 January-March 2015

Coccomyces prominens sp. nov. (Rhytismataceae)

on Rhododendron coeloneurum in China

YA-FEI Xu’, YUAN Wu’, YAN-QIONG MENG’,

SHI-JUAN WANG?, & YING-REN LIN”

' School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

ABSTRACT — A fungus found on leaves of Rhododendron coeloneurum from Garze prefecture

(Sichuan Province), China, is described as Coccomyces prominens. The new species is similar

to C. urceoloides but differs by its much larger intraepidermal ascomata and the presence of

conidiomata. The type specimen is deposited in the Reference Collection of Forest Fungi of

Anhui Agricultural University, China (AAUF).

Key worps — foliicolous fungus, morphological character, taxonomy, Ericaceae

Introduction

Coccomyces De Not. is the second-largest genus of Rhytismataceae

(Rhytismatales, Leotiomycetes, Ascomycota), represented by 116 species recorded

by Kirk et al. (2008) plus a few recently added. Members of this genus are

widely distributed and occur on leaves, twigs, bark, or wood of vascular plants,

especially Ericaceae, Fagaceae, and Lauraceae (Sherwood 1980). Twenty-eight

Coccomyces species have been reported from China (Korf & Zhuang 1985; Lin

1998; Hou et al. 2006, 2007; Jia et al. 2012; Zheng et al. 2012; Wang et al. 2013;

Yang et al. 2013) since Teng (1934) first recorded C. dentatus (J.C. Schmidt)

Sacc. on Quercus and Castanea and C. delta (Kunze) Sacc. on Lauraceae.

Here, Coccomyces prominens on leaves of Rhododendron coeloneurum from

Luding county, Garze prefecture, Sichuan Province, China, is described as a

new species.

Materials & methods

Mature fruiting bodies were selected from the collected specimens. External shapes,

size, color, ascomatal and conidiomatal openings, and zone lines were observed under

74 ... Xu & al.

a dissecting microscope at 10—-50x magnification. After rehydration in water, 10-20

tum thick vertical transverse sections of fruiting bodies were made using a freezing

microtome and mounted in 0.7% (w/v) cotton blue in water. Gelatinous sheaths

surrounding ascospores and paraphyses were observed in water or 0.1% (w/v) cotton

blue in phenol glycerin. The colors of internal structures and paraphysis contents were

observed in water. Measurements were made using material in 5% KOH and from ca

30 paraphyses, asci, ascospores, conidiogenous cells, and conidia for each specimen.

Figures were drawn of the external and internal structures of the fruiting bodies using a

microscopic drawing tube. The type collection is deposited in the Reference Collection

of Forest Fungi of Anhui Agricultural University, Hefei, China (AAUF).

Taxonomy

Coccomyces prominens Y.F. Xu & Y.R. Lin, sp. nov. Fics 1-6

MycoBank MB 808960

Differs from Coccomyces urceoloides by its much larger, intraepidermal ascomata with

an obvious pre-formed dehiscence mechanism, its wider ascospores enveloped in a

gelatinous sheath, and its production of conidiomata.

Type: China, Sichuan, Garze prefecture, Luding, Hailuogou, alt. ca 3400m, on fallen

leaves of Rhododendron coeloneurum Diels (Ericaceae), 22 June 2009, Y.G. Liu & Y.R. Lin

2768 (Holotype, AAUF 68876).

ETYMOLOGY: prominens (Latin = prominent), referring to the ascomata, which are

noticeable on the substratum.

ZONE LINES usually frequent, brown or dark brown, thin to wide, somewhat

diffused, nearly always partly surrounding the bleached spots.

ConipiomMara in similar positions to ascomata on the substratum, scattered

to clustered, sometimes several coalescent. In surface view, conidiomata

150-220 um diam., rounded or subrounded, with a clearly marked outline,

black-brown, moderately raising the leaf surface, discharging spores through

an apical ostiole. In vertical transverse section, conidiomata intraepidermal,

lenticular to semicircular. UPPER WALL 6.5—12 um thick, almost not attenuating

towards the base, composed of disparately sized angular cells. BASAL WALL

35-50 um thick in the middle, becoming 13-20 um thick at periphery,

consisting of black-brown, angular and elongate, thick-walled cells 3.5-8 um

diam. SUBCONIDIOGENOUS LAYER 5-8 um thick, composed of colorless, thin-

walled angular cells 1.2-2 um diam. CONIDIOGENOUS CELLS 10-14 x 1.5-2.2

um, cylindrical but tapered towards the apex, hyaline, proliferating sympodially

and percurrently. Conrp1a 4.5-7 x 0.5-0.8 um, cylindrical, straight or slightly

curved to one side, ends rounded or bluntly pointed, hyaline, aseptate, smooth-

walled. TRICHOGYNES 35-45 x 2-3 um, cylindrical, tapering towards the apex,

1—2-septate near the base.

Ascomarta epiphyllous, scattered or occasionally coalescent, in subcircular

or irregular pale areas with obvious edges. In surface view, ascomata 800-1400

Fs)

Coccomyces prominens sp. nov. (China) ...

Pee

ory

6. Coccomyces prominens (ex holotype, AAUF 68876) on Rhododendron coeloneurum.

Fics 1

and a zone line observed

ascomata,

under a dissecting microscope. 3. Ascoma in vertical transverse section. 4. Portion of ascoma in

1. A leaf bearing fruiting bodies and zone lines. 2. Conidiomata,

asci, and ascospores. 6. Conidioma in vertical transverse

vertical transverse section. 5. Paraphyses,

section.

76 ... Xu &al.

um diam., quadrilateral to octagonal, black-brown to black, not shiny or

slightly shiny, edge defined, moderately raising the substrate surface, with

an obvious pre-formed dehiscence mechanism, opening by 4-8 radial splits

nearly extend to the edge of ascoma. Lips absent. In median vertical transverse

section, ascomata intraepidermal. COVERING sTROMA 55-65 um thick near

the opening, decreasing to 25-30 um thick adjacent to the base, composed

of black-brown textura angularis-epidermoidea with thick-walled cells 4—6.5

um diam., connecting to the basal stroma. Adjacent to the top of the covering

stroma, a flimsy structure develops consisting of angular, thin-walled, almost

colorless cells staining with cotton blue. BasaL stroma 18-22 um thick,

comprised of 2—3(—4) layers of dark brown, angular, thick-walled cells 4-6 um

diam. ExcrpuLUM well-developed, 45-60 um thick, arising from the marginal

paraphyses and the internal matrix of stroma, consisting of closely agglutinated

septate hyphae 2—5 um diam., mostly short-celled, with swollen greyish-brown

apices. INTERNAL MATRIX OF STROMA moderately developed, 20-50 um thick,

comprising hyaline, gelatinised textura intricata. SUBHYMENIUM colorless,

10-15 um thick, composed of textura angularis. PARAPHYSES 160-190 x

1.8—2.2 um, filiform, 0-1-septate, occasionally branched, yellowish-brown

in the upper quarter, surrounded by a 0.6—0.8 um thick gelatinous matrix,

gradually irregularly swollen to 3-5 um and containing disparately-sized oil

drops near the apex, forming a 35-50 um thick epithecium inserted with a

number of hyaline, refractive, sub-oblong solid gels 12-32 x 8-14 um. AscI

ripening sequentially, 110-170 x 12—15 um, cylindrical-clavate, short-stalked,

thin-walled, apex bluntly pointed to subtruncate-conical, J-, 8-spored.

ASCOSPORES arranged fasciculately, 80-120 x 2-2.4 um, filiform, hyaline,

aseptate, enveloped in a 0.6—0.9 um thick gelatinous sheath.

HOST SPECIES, HABITAT AND DISTRIBUTION: Producing ascomata and

conidiomata on fallen leaves of Rhododendron coeloneurum. Known only from

the type locality, Sichuan Province, China.

COMMENTS — Coccomyces prominens resembles C. urceoloides Spooner in

the well-developed excipulum with more or less brown hyphal tips, gradually

swollen paraphyses at the apex, and refractive solid gels in the epithecium.

However, C. urceoloides has smaller, hypophyllous, subepidermal ascomata

(350-550 um diam.) not associated with conidiomata; much smaller, narrowly

cylindrical asci (120-135 x 5.5—-6 um); and smaller ascospores (80-98 x ca 1

um) with a mucilaginous cap at the proximal end (Spooner 1990).

Coccomyces sinensis Y.R. Lin & Z.Z. Li is easily distinguished from

C. prominens by its triangular to pentagonal ascomata, smaller asci (108-130 x

5.3—-6.0 um), smaller ascospores (54—90 x 1—1.2 um), subhymenium consisting

of textura porrecta-intricata, and distinct paraphyses that are abruptly swollen

Coccomyces prominens sp. nov. (China) ... 77

to subfusoid-ventricose with a subcylindrical mucro near the apex (Lin et al.

2001).

The similar C. monticola Sherwood differs in the smaller and orbicular

ascomata (500-800 um diam.), a thinner covering stroma (20 um thick),

slightly swollen paraphysial apices (2.5 um wide), and much narrower asci (8-9

um wide; Sherwood 1980).

Acknowledgements

We are grateful to Dr J.E. Taylor and Dr M. Ye for serving as pre-submission

reviewers, and to Dr Y.G. Liu for the field investigations. This study was supported by

the National Natural Science Foundation of China (No. 31270065, 31170019).

Literature cited

Hou CL, Piepenbring M. 2007. Two new species of Rhytismataceae on twigs of conifers from

Yunnan Province, China. Mycotaxon 102: 165-170.

Hou CL, Kirschner R, Chen CJ. 2006. A new species and new records of Rhytismatales from

Taiwan. Mycotaxon 95: 71-79.

Jia GJ, Lin YR, Hou CL. 2012 [“2011”]. A new species of Coccomyces (Rhytismatales, Ascomycota)

from Mt. Huangshan, China. Mycotaxon 118: 231-235. http://dx.doi.org/10.5248/118.231

Kirk PM, Cannon PF, Minter DW, Stalpers JA (eds). 2008. Dictionary of the fungi, 10th ed. CAB

International. Wallingford. 771 p.

Korf RP, Zhuang WY. 1985. Some new species and new records of Discomycetes in China.

Mycotaxon 22: 483-514.

Lin YR. 1998. Studies on Coccomyces de Notaris and Neococcomyces gen. nov. in China. [Ph.D.

thesis; in Chinese]. Northeast Forestry University. Harbin. 98 p.

Lin YR, Li ZZ, Xu ZS, Wang JR, Yu SM. 2001. Studies on the genus Coccomyces from China IV

[in Chinese]. Mycosystema 20: 1-7.

Sherwood MA. 1980. Taxonomic studies in the Phacidiales: The genus Coccomyces (Rhytismataceae).

Occasional Papers of the Farlow Herbarium of Cryptogamic Botany 15: 1-120.

Spooner BM. 1990. Coccomyces and Propolis (Rhytismatales) from Mt Kinabalu, Borneo. Kew

Bulletin 45: 451-484. http://dx.doi.org/10.2307/4110513

Teng SC. 1934. Notes on Discomycetes from China. Sinensia 5: 431—465.

Wang SJ, Lin YR, Tang YP, Li K. 2013 [“2012”]. A new species of Coccomyces (Rhytismatales,

Ascomycota) on Ilex elmerrilliana. Mycotaxon 122: 287-291. http://dx.doi.org/10.5248/122.287

Yang MS, Lin YR, Zhang L, Wang XY. 2013 [“2012”]. Coccomyces hubeiensis, a new fungus of

Rhytismatales from China. Mycotaxon 122: 249-253. http://dx.doi.org/10.5248/122.249

Zheng Q, Lin YR, Yu SM, Chen L. 2012 [“2011”]. Species of Rhytismataceae on Lithocarpus spp.

from Mt Huangshan, China. Mycotaxon 118: 311-323. http://dx.doi.org/10.5248/118.311

MY COTAXON

http://dx.doi.org/10.5248/130.79

Volume 130, pp. 79-89 January-March 2015

Type studies on Amauroderma species described by

J.D. Zhao et al. and the phylogeny of species in China

MENG-JIE Li”? & Ha1l-SHENG YUAN”

"State Key Laboratory of Forest and Soil Ecology, Institute of Applied Ecology,

Chinese Academy of Sciences, Shenyang 110164, P. R. China

? University of Chinese Academy of Sciences, Beijing 100049, P. R. China

* CORRESPONDENCE TO: hsyuan@iae.ac.cn

ABSTRACT — Holotypes of eight Amauroderma species described by Ji-Ding Zhao and

co-workers, together with 93 additional specimens collected from China, were examined.

Four names are demoted to synonymy (A. amoiense and A. wuzhishanense = A. rugosum;

A. dayaoshanense = Pyrrhoderma sendaiense; A. fujianense = Ganoderma fornicatum), two

represent accepted species (A. austrosinense, A. yunnanense), and two are of uncertain

application (A. longgangense, A. jiangxiense) because of the poor or sterile condition of the

holotype material. Illustrated descriptions of A. austrosinense and A. yunnanense are given

based on the holotypes. Four other species reported from China (A. perplexum, A. rugosum,

A. subresinosum, A. yunnanense) were sequenced for molecular analysis, and a phylogenetic

tree was constructed by maximum parsimony and Bayesian analyses of the nuclear

internal transcribed spacer of ribosomal DNA (ITS rDNA) sequences. A key to accepted

Amauroderma species in China is provided.

KEY worDs — Ganodermataceae, morphology, phylogenetics, taxonomy

Introduction

Amauroderma Murrill was established by Murrill and typified by A.

regulicolor (Berk. ex Cooke) Murrill (Murrill 1905). This genus is mainly

characterized by sessile or stipitate (mesopodal, pleuropodal) basidiocarps

with a laccate or dull pileus; a trimitic hyphal system; and ellipsoid, subglobose

to globose bitunicate basidiospores with an asperulate inner wall (Furtado

1981, Ryvarden 2004). Macroscopically, Amauroderma shares with Ganoderma

P. Karst. a similar basidiocarp shape of central or lateral stipe and a laccate or

dull surface. Ganoderma can be separated from Amauroderma by its distinctly

truncate basidiospores, and most Ganoderma species grow on dead wood,

while most Amauroderma species grow in the ground from buried roots

(Ryvarden 2004).

80 ... Li & Yuan

Amauroderma is widely distributed in the tropics and subtropics south of

25°N in China within Guangdong, Guangxi, Yunnan, and Hainan Provinces.

Amauroderma species are regarded as economically valuable because of their

famous medicinal properties and pathogenicity (Dai et al. 2007, 2009; Jiao et al.

2013; Chan et al. 2013). Index Fungorum (www.indexfungorum.org) has listed

126 names under this genus. In China, taxonomic research on Amauroderma

began with Teng, who recorded 10 species (Teng 1936, 1939, 1963). Later Zhao

et al. (1979, 1983, 1984) and Zhao & Zhang (1986, 1987a,b, 2000) reported

20 species, nine of which were new. The types of these new species [except

Amauroderma guangxiense J.D. Zhao & X.Q. Zhang (not traced)] are preserved

in the Herbarium Mycologicum Academiae Sinicae (HMAS), the Institute of

Microbiology, Chinese Academy of Sciences. Tai (1979) and Fan & Liu (1990)

have also contributed to Amauroderma records in China.

In this study we investigated the type specimens of Amauroderma described

by Ji-Ding Zhao and his colleagues and 93 other recently collected specimens

from southern China based on morphological characters and rDNA sequence

analyses. The taxa are presented below in alphabetical order according to the

original names.

Materials and Methods

Morphology

The studied specimens were deposited in the Herbaria of the Institute of Applied

Ecology, Chinese Academy of Sciences (IFP); the Institute of Microbiology, Chinese

Academy of Sciences (HMAS); and the Institute of Microbiology, Beijing Forestry

University (BJFC). Microscopic procedures followed He & Dai (2012). Sections were

studied at magnifications up to 1000x using a Nikon Eclipse 80i microscope with phase

contrast illumination. Measurements were made from sections stained with Cotton

Blue. In presenting basidiospore size variation, 5% of measurements are excluded

from each end of the range and given in parentheses. The following abbreviations are

used: CB = Cotton Blue, CB+ = cyanophilous, 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 = variation in the L/W ratios between

the specimens studied, and n = number of basidiospores measured from given number

of specimens. Special color terms follow Petersen (1996).

Molecular phylogeny

Amplification of ITS sequences from herbarium specimens was conducted using

Phire® Plant Direct PCR Kit (Finnzymes, Finland) with ITS5 and ITS4 primers (White et

al. 1990). PCR procedure was as follows: initial denaturation at 98 °C for 5 min, followed

by 39 cycles at 98 °C for 5 s, 59 °C for 10 s and 72 °C for 5 s, and a final extension at

72 °C for 2 min. PCR amplification was confirmed on 1% agarose electrophoresis gels

Amauroderma type studies (China) ... 81

TABLE 1. ITS sequences of Amauroderma and Tomophagus species used in the

molecular analyses.

SPECIES VOUCHER LOCALITY GENBANK No.

A. perplexum Cui 6496 Hainan, China KJ531650

Dai 10811 Hainan, China KJ531651

Wei 5562 Hainan, China KJ531652

A. rugosum Cui 6285 Hainan, China KJ531656

Zhou 153 Guangxi, China KJ531657

Dai 7862 Hainan, China KJ531658

Zhou 136 Guangxi, China KJ531659

Zhou 347 Guangxi, China KJ531660

Zhou 66 Guangxi, China KJ531661

Yuan 4554 Hainan, China KJ531662

Dai 9904 Hainan, China KJ531663

Cui 9011 Guangdong, China KJ531664

Cui 9012 Guangdong, China KJ531665

Wei 5234 Hainan, China KJ531666

Cui 8882 Guangdong, China KJ531667

Dai 9553 Hainan, China KJ531668

Dai 9566 Hainan, China KJ531669

Dai 4345 Hainan, China KJ531670

Dai 10040 Hainan, China KJ531671

Cui 4078 Fujian, China KJ531672

Dai 10307 Hainan, China KJ531673

Zhou 523 Guangxi, China KJ531674

Zhou 547 Guangxi, China KJ531675

Dai 12324 Yunnan, China KJ531676

Dai 12390 Yunnan, China KJ531677

Cui 9006 Guangdong, China KJ531678

THP 30 Unknown HM 480835

A. subresinosum Wei 5569 Hainan, China KJ531649

ML 288 Malaysia JQ409358

A. yunnanense Cui 7974 Yunnan, China KJ531653

Dai 13021 Yunnan, China KJ531654

Yuan 2253 Yunnan, China KJ531655

T. colossus CGMCC 5.763 Philippines JQ081068

stained with ethidium bromide (Stéger et al. 2006). The PCR products were purified and

directly sequenced in Beijing Genomics Institute, China, with the same primers. The

most similar sequences (TABLE 1) were retrieved from GenBank (http://www.ncbi.nlm.

gov) using the BLAST option. Sequences were aligned using ClustalX (Thompson et al.

82 ... Li & Yuan

1997). Alignment was manually adjusted to allow maximum alignment and minimize

gaps, and deposited in TreeBASE (http://treebase.org/treebase-web/; submission ID:

17050). Identity/similarity between two sequences was calculated using the “pairwise

alignment, calculation of the similarity/identity” option of BioEdit v. 7.0.5 (Hall 2005).

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 (PAUP* version 4.0b10) was used (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 auto-increase, branches of zero length were collapsed,

and all parsimonious trees were saved. Clade stability was assessed using a bootstrap (BT)

analysis with 1,000 replicates (Felsenstein 1985). Descriptive tree statistics, including

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.

JModelTest (Posada 2008) was used to determine the best-fit evolution model

for each data set for Bayesian inference (BY). Bayesian inference was calculated with

MrBayes3.1.2 with a general time reversible (GTR) model of DNA substitution and a

gamma distribution rate variation across sites (Ronquist & Huelsenbeck 2003). Four

simultaneous Markov chains were run with 3 million generations, and trees were

sampled every 100 generations. Trees generated for the first one-fourth generations

were discarded as burn-in. A 50% majority rule consensus tree of all remaining trees

was calculated. Branches that received maximum parsimony bootstrap support (BSS)

and Bayesian posterior probabilities (BPP) respectively greater than or equal to 75% and

0.95 were considered as significantly supported.

Taxonomy

We studied eight species of Amauroderma (Ganodermataceae) described

from China by Ji-Ding Zhao et al. Four of the eight names are synonymized,

two are accepted, and two are of uncertain application.

Amauroderma amoiense J.D. Zhao & L.W. Hsu, Acta Mycol. Sinica 2: 164. 1983.

Ho.otype: China. Fujian Prov., Xiamen, sandy land, 13.V1.1976, Li Hui-zhong &

Zhang Xiao-qing 28 (HMAS 42784).

= Amauroderma rugosum (Blume & T. Nees) Torrend, Brotéria, Sér. Bot. 18: 127. 1920.

The type material is sterile, but all other characters fit A. rugosum well. For a

detailed description of A. rugosum, see Ryvarden & Johansen (1980).

Amauroderma austrosinense J.D. Zhao & L.W. Hsu, Acta Mycol. Sinica

3: 20. 1984. Fig 1

Hototype: China. Hainan Prov., Bawangling, ground, 20.1V.1977, Han Shu-jin 902

(HMAS 42695).

BASIDIOCARPS annual, stipitate, umbelliform. PILEus single, suborbicular to

circular, glabrous, dull, clay-buff to fulvous brown, densely concentrically zoned

Amauroderma type studies (China) ... 83

Fic. 1. Amauroderma austrosinense (holotype): basidiospores.

and slightly irregularly radially furrowed; MARGIN obtuse and deflexed when

dry; STIPE sub-central or eccentric, concolorous with pileus, cavitate, up to 12

cm long and 1 cm thick. CONTEXT cinnamon, dense and hard, homogenous, up

to 4mm thick. PORE SURFACE cream white when fresh, turning brown with age;

PORES circular, entire, 6-8 per mm. TUBES woody, up to 3 mm long.

HYPHAL SYSTEM trimitic; GENERATIVE HYPHAE with clamp connections,

hyaline, thin-walled, 2.8-4 um in diam; SKELETAL HYPHAE pale yellow to almost

colorless, sparsely branched, 3.7-5.2 um in diam; BINDING HYPHAE hyaline,

branched, tortuous, 1.5-2.2 um in diam; all the hyphae IKI-, CB+; tissues

darkening in KOH. Basipiospores subglobose to globose, faintly yellowish,

IKI-, CB+, doubled-walled, exospore smooth, endospore with conspicuous

echinule, (6.5-)6.8-7.6(-8) x (5.9-)6.2-7(-7.3) um, L = 7.34 um, W = 6.58 um,

Q = 1.12 (n = 30/1).

ADDITIONAL SPECIMEN EXAMINED: CHINA, HAINAN PRov., Changjiang County,

Bawangling Nat. Res., ground, 20.IV.1977 (HMAS 42695).

REMARKS: Amauroderma austrosinense is an accepted species, remarkable for

its umbelliform basidiocarps with distinct concentric rings, small pores, and

basidiospores. A. camerarium (Berk.) J.S. Furtado is similar to A. austrosinense

by sharing a clay-colored, densely concentrically zoned, and radially furrowed

pileus. However, A. camerarium differs in its larger basidiospores (12-15 x

10-13 um; Ryvarden 2004).

Amauroderma leucosporum Corner may be confused with A. austrosinense

by having similar-sized subglobose to globose basidiospores and also not

turning blood red on bruising, but A. leucosporum can be distinguished by its

finely to distinctly villous pileus and stipe (Corner 1983).

84 ... Li & Yuan

Amauroderma dayaoshanense J.D. Zhao & X.Q. Zhang, Acta Mycol. Sinica

6551987

Ho .ortyPe: China. Guangxi Auto. Reg., Dayaoshan, dead wood, 1982, Wei Bing-gang

70 (HMAS 48282).

= Pyrrhoderma sendaiense (Yasuda) Imazeki, Trans. Mycol. Soc. Japan 7: 4. 1966.

The type is sterile but has the characteristics of P sendaiense (see Nunez &

Ryvarden (2000) for a detailed description).

Amauroderma fujianense J.D. Zhao, L.W. Hsu & X.Q. Zhang, Acta Microbiol. Sinica

T9279. 1979:

Ho.otype: China. Fujian Prov., Sanming County, rotten wood, 28.VI.1976, Zhang

Xiao-qing 68 (HMAS 37919).

= Ganoderma fornicatum (Fr.) Pat., Bull. Soc. Mycol. Fr. 5: 71. 1889.

The holotype is sterile but represents a juvenile specimen of G. fornicatum (see

Patouillard (1889) for a detailed description).

Amauroderma jiangxiense J.D. Zhao & X.Q. Zhang, Acta Mycol. Sinica

6: 206. 1987.

Ho .ortyPe: China. Jiangxi Prov., Jian County, rotten wood, 15.11.1983, Yang Ren-gen

152 (HMAS 50418).

The basidiocarp is umbelliform and the pileus is black, laccate, and shiny, but

the specimen is completely sterile. It probably represents a Ganoderma species,

but the application of the name is uncertain.

Amauroderma longgangense J.D. Zhao & X.Q. Zhang, Acta Mycol. Sinica

Sr227 L986,

Ho.otype: China. Guangxi Auto. Reg., Nonggang, on rotten wood, 1979, Wei Bing-

gang 790111-G8 (HMAS 47621).

Basidiospores subglobose, 6-8.5 x 5-8 um, with spinules 0.8-1.2 um on

sporoderm. However, the single-walled basidiospore obviously differs from

the double-walled spore of other Amauroderma species. The application of the

name is uncertain.

Amauroderma wuzhishanense J.D. Zhao & X.Q. Zhang, Acta Mycol. Sinica

6: 208. 1987.

Ho.otype: China. Hainan Prov., Wuzhishan, on rotten wood, 22.VII.1956, Jiang

Guang-zheng (HMAS 19311).

= Amauroderma rugosum (Blume & T. Nees) Torrend, Brotéria, Sér. Bot. 18: 127. 1920.

The holotype is in accordance with Amauroderma rugosum, with its pileus

obscurely zonate, radially quite rugose, and warted. For a detailed description

of A. rugosum, see Ryvarden & Johansen (1980).

Amauroderma type studies (China) ... 85

Fic. 2. Amauroderma yunnanense (holotype): basidiospores.

Amauroderma yunnanense J.D. Zhao & X.Q. Zhang, Acta Mycol. Sinica,

Suppl. 1: 268. 1987 [“1986”]. FIG 2

Ho.otype: China. Yunnan Prov., Xichou County, ground, Xiaogiaogou, 14.V.1959,

Wang Qing-zhi 89 (HMAS 48231).

BASIDIOCARPS annual, centrally to laterally stipitate, coriaceous to corky,

brittle when dry. PiLeus single, flabelliform, suborbicular, umbilicate, PILEAL

SURFACE pale yellowish brown to dark brown, adpressed velutinate, with distinct

concentric zones at the margin and fine furrows in the middle; MARGIN inflexed,

acute or subacute, entire or slightly lacerated and thin; stipe concolorous with

the pileus, up to 7 cm long, inflated at base. ConTExT homogeneous, white

to pale yellow. PoRE SURFACE white when fresh, pale straw yellow when dry,

bruising unchanged; porgs angular, 2-3 per mm; TuBEs concolorous with pore

surface, up to 5 mm thick.

HYPHAL SYSTEM trimitic; GENERATIVE HYPHAE with clamps, hyaline,

thin-walled, 2-4.3 um in diam; SKELETAL HYPHAE thick-walled, pale yellow,

arboriform, 3.5-5.5 um in diam; BINDING HYPHAE hyaline, branched, tortuous,

1.8-2.5 um in diam; all the hyphae IKI-, CB+; tissues darkening in KOH.

Basipiospores broadly ellipsoid to subglobose, pale yellow brown, IKI-,

CB+, doubled-walled, exospore smooth, hyaline, endospore with conspicuous

echinule, (8.3—)8.9-10.3(-10.7) x (6.3-)6.7-8(-8.8) um, L = 9.35 um, W = 7.32

um, Q = 1.22-1.34 (n = 120/4).

ADDITIONAL SPECIMENS EXAMINED: CHINA, YUNNAN PRov., Chuxiong, Zixishan

Forest Park, ground, 8.[X.2006, Yuan 2253 (IFP 013207); Ailaoshan, 5.[X.2007, Yuan

3367 (IFP 013182); Kunming, Qiongzhusi Park, 21.X.2009, Cui 7974 (BJFC 006463);

Xiaoshao Village, 22.1X.2012, Dai 13021 (IFP 019123).

REMARKS: Amauroderma yunnanense is an accepted species characterized by a

brown tomentose pileus and stipe.

Amauroderma perplexum Corner and A. yunnanense both have a velutinate

pileus and stipe as well as stipes that dilate at the base; A. perplexum can be

86 ... Li & Yuan

separated by larger basidiospores, longer tubes, and a pore surface that changes

color when bruised (Corner 1983).

Amauroderma partitum (Berk.) Wakef., which may be confused with

A. yunnanense as they share similar angular pores that are 2-3 per mm, can

be distinguished by its glabrous, slightly shiny pileus and larger basidiospores

(12-15 x 8-10 um; Ryvarden 2004).

Molecular phylogeny

The nuclear ITS rDNA dataset contains 32 sequences representing

A. perplexum, A. rugosum, A. subresinosum (Murrill) Corner, and A. yunnanense,

with Tomophagus colossus (Fr.) Murrill as outgroup (TasBLe 1). After the

ambiguous sites at both ends and the 5.8S region were trimmed off, the

alignment comprised 561 characters, of which 462 were constant, 22 were

variable but parsimony-uninformative, and 77 were parsimony-informative.

Maximum parsimony analysis yielded 6 equally parsimonious trees

(CI = 0.918, RI = 0.952, RC = 0.874, HI = 0.082). A strict consensus tree of these

trees is presented in Figure 3. The 50% majority consensus tree generated by

the Bayesian analysis showed a similar topology with the strict consensus MP

tree, and only the topology from MP analysis is presented, while both bootstrap

support value (BSS) and Bayesian posterior probabilities (BPP) are shown at

the nodes (Fic. 3).

In the phylogenetic tree, each sampled Amauroderma species was resolved

as a strongly supported clade (100% BSS and 1.00 BPP). Identity/similarity

calculation result shows that identities of Amauroderma sequences are

between 0.98 and 1. We failed to obtain sequences from the type specimens of

Amauroderma described by Ji- Ding Zhao from China as the materials were too

old to extract integrated DNA.

Key to species of Amauroderma in China

i. Pileal surface appressed welutinate:.o iiss talee ge hues Aol h eva heh Wale Thee 2

Te Pileal ‘surface: PlaDEOUS.§ 4p seven aybscton teh acbea dring.seg rhea son dhe soa dle puon toby baud he fobs deh fro 9 3

2. Pores 2-3 per mm, pore surface straw yellow, not reddening on bruising,

spores (8.3—)8.9-10.3(-10.7) x (6.3-)6.7-8(-8.8) um. .......... A. yunnanense

2. Pores 4-6 per mm, pore surface cream white, reddening on bruising,

spores (10.3-)10.8-14(-14.8) x (8.9-)9.2-12(-13.1) um......... A. perplexum

3. Pileus surface laccate, spores ovoid to broadly ellipsoid,

(13.7-)14.6-17.1(-17.5) x (8.3-)8.9-10.2(-11.3) um.......... A. subresinosum

SaPileal Sut acevcUlle., £ reste dove. tdan ove len aortas Sahy eeselger estes ace aeearatenat hts gt mmaie ates -:

4, Pileal clay-buff to fulvous brown, pores 7-8 per mm,

spores (6.5—)6.8-7.6(-8) x (5.9-)6.2-7(-7.3) um .........0.00. A. austrosinense

4. Pileus taupe, dark brown to black, pores 4-6 per mm,

spores (8.1—)8.4-11.7 (-12) x (7-)7.5-9.6(-10.3) um. ............. A. rugosum

Amauroderma type studies (China) ... 87

100/1.00 rAmauroderma subresinosum Wei 5569

Amauroderma subresinosum JQ 409358

Amauroderma yunnanense Cui 7974

ROUEOe Amauroderma yunnanense Dai 13021

100/1.00 Amauroderma yunnanense Yuan 2253

Amauroderma perplexum Cui 6496

100/08 Amauroderma perplexum Wei 5562

‘Amauroderma perplexum Dai 10811

Amauroderma rugosum Cui 6285

52/0.52 Amauroderma rugosum Zhou 153

Amauroderma rugosum Dai 7862

Amauroderma rugosum Zhou 136

Amauroderma rugosum Zhou 347

Amauroderma rugosum Zhou 66

62/0.97 Amauroderma rugosum Yuan 4554

Amauroderma rugosum Dai 9904

Amauroderma rugosum HM 480835

Amauroderma rugosum Cui 9011

Amauroderma rugosum Cui 9012

Amauroderma rugosum Wei 5234

Amauroderma rugosum Cui 8882

100/1.00 Amauroderma rugosum Dat 9553

Amauroderma rugosum Dai 9566

Amauroderma rugosum Dai 4345

Amauroderma rugosum Dai 10040

Amauroderma rugosum Cui 4078

Amauroderma rugosum Dai 10307

Amauroderma rugosum Zhou 523

Amauroderma rugosum Zhou 547

Amauroderma rugosum Dai 12324

Amauroderma rugosum Dai 12390

Amauroderma rugosum Cui 9006

Tomophagus colossus JQ 081068

Liooorsiets

0.0 8.0

Fic. 3. Strict consensus tree obtained from Maximum Parsimony and Bayesian analyses of ITS

sequences of Amauroderma. Tomophagus colossus was used as outgroup. Numbers at branches

indicate parsimony bootstrap values and Bayesian posterior probabilities values higher than 50%.

88 ... Li & Yuan

Acknowledgements

The authors are grateful for presubmission review and helpful comments by Bao-

Kai Cui (Institute of Microbiology, Beijing Forestry University) and Xiao-Yong Liu

(Institute of Microbiology, Chinese Academy of Sciences). Special thanks are due to Dr.

Katherine F. LoBuglio (Harvard University Herbaria, USA) for checking and improving

the manuscript. This research was financed by the National Natural Science Foundation

of China (Project Nos. 31170022 & 31470148).

Literature cited

Chan PM, Kanagasabapathy G, Tan YS, Sabaratnam V, Kuppusamy UR. 2013. Amauroderma

rugosum (Blume & T. Nees) Torrend: nutritional composition and antioxidant and potential

anti-inflammatory properties. Evid.-Based Compl. Alt. Med. 2013, Article ID 304713, 10 pages.

http://dx.doi.org/10.1155/2013/304713

Corner EJH. 1983. Ad Polyporaceas I. Amauroderma and Ganoderma. Beheift. Nova Hedwigia 75.

183 p.

Dai YC, Cui BK, Yuan HS, Li BD. 2007. Pathogenic wood-decaying fungi in China. Forest Pathol.

37:105-120. http://dx.doi.org/10.1111/j.1439-0329.2007.00485.x

Dai YC, Yang ZL, Cui BK, Yu CJ, Zhou LW. 2009. Species diversity and utilization of medicinal

mushrooms and fungi in China. Int. J. Med. Mushr. 11: 287-302.

http://dx.doi.org/10.1615/Int)MedMushr.v11.i3.80

Fan L, Liu B. 1990. A new species of Amauroderma. Acta Micol. Sinica 9: 202-205.

Felsenstein J. 1985. Confidence intervals on phylogenetics: an approach using bootstrap. Evolution

39: 783-791.

Furtado JS. 1981. Taxonomy of Amauroderma (Basidiomycetes, Polyporaceae). Mem. New York

Bot. Gard. 34. 109 p.

Hall T. 2005. BioEdit: biological sequence alignment editor for Win95/98/NT/2 K/XP. Ibis

Therapeutic, Carlsbad, CA.

He SH, Dai YC. 2012. Taxonomy and phylogeny of Hymenochaete and allied genera of

Hymenochaetaceae (Basidiomycota) in China. Fungal Divers. 56: 77-93.

Jiao CW, Xie YZ, Yang XL, Li HR, Li XM, Pan HH, Cai MH, Zhong HM, Yang BB. 2013. Anticancer

activity of Amauroderma rude. PLOS ONE 8(6): e66504.

http://dx.doi.org/10.1371/journal.pone.0066504

Murrill MA. 1905. The Polyporaceae of North America XI. A synopsis of the brown pileate species.

Bull. Torrey Bot. Club 32: 353-371.

Nujez M, Ryvarden L. 2000. East Asian polypores. Synopsis Fungorum 13. 168 p.

Patouillard NT. 1889. Le genre Ganoderma. Bulletin de la Société Mycologique de France 5: 64-80.

Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve.

Posada D. 2008. jModelTest: Phylogenetic model averaging. Mol. Biol. Evol. 25: 1253-1256.

http://dx.doi.org/10.1093/molbev/msn083

Ronquist F, Huelsenbeck JP. 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. http://dx.doi.org/10.1093/bioinformatics/btg180

Ryvarden L. 2004. Neotropical polypores Part 1. Syn. Fung. 19: 38-69.

Ryvarden L, Johansen I. 1980. A preliminary polypore flora of East Africa. Fungiflora, Oslo.

Stdger A, Schaffer J, RuppitschW. 2006. A rapid and sensitive method for direct detection of Erwinia

amylovora in symptomatic and asymptomatic plant tissues by polymerase chain reaction.

J. Phytopathol. 154: 469-473.

Amauroderma type studies (China) ... 89

Swofford DL. 2002. PAUP*. Phylogenetic analysis using parsimony (*and other methods). Version

4.0b10. Sinauer Associates, Sunderland, Massachusetts.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Peking (in Chinese).

Teng SC. 1936. Additional fungi from China III. Sinensia. 7:529-569.

Teng SC. 1939. A contribution to our knowledge of the higher fungi of China. National Institute of

Zoology & Botany, Academia Sinica.

Teng SC. 1963. Fungi of China. Science Press, Beijing (in Chinese).

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal_X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Res. 25:876-4882.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis MA et al. (eds). PCR protocols, a guide to

methods and applications. Academic, San Diego.

Zhao JD, Zhang XQ. 1986. Taxonomic studies on Ganodermataceae of China V. Acta Mycol. Sinica

Br 219F2 25;

Zhao JD, Zhang XQ. 1987a [“1986”]. Taxonomic studies on Ganodermataceae of China VII - the

genus Amauroderma. Acta Mycol. Sinica, Suppl. 1: 258-272.

Zhao JD, Zhang XQ. 1987b. Taxonomic studies on Ganodermataceae of China VII. Acta Mycol.

Sinica 6: 199-210.

Zhao JD, Zhang XQ. 2000. Flora Fungorum Sinicorum, vol. 18. Ganodermataceae (in Chinese).

Sciences Press, Beijing.

Zhao JD, Xu LW, Zhang XQ. 1979. Taxonomic studies on the subfamily Ganodermoideae of China

(in Chinese). Acta Microbiol. Sinica 19: 265-279.

Zhao JD, Xu LW, Zhang XQ. 1983. Taxonomic studies on Ganodermataceae of China II. Acta

Mycol. Sinica 2: 159-167.

Zhao JD, Xu LW, Zhang XQ. 1984. Taxonomic studies on Ganodermataceae of China III. Acta

Mycol. Sinica 3: 15-23.

MY COTAXON

http://dx.doi.org/10.5248/130.91

Volume 130, pp. 91-103 January-March 2015

Cladonia corymbescens consists of two species

TEUVO AHTI, RAQUEL PINO-BODAS, & SOILI STENROOS

Botanical Museum, Finnish Museum of Natural History, University of Helsinki,

PO. Box 7, FI-00014 Helsinki, Finland

* CORRESPONDENCE TO: teuvo.ahti@helsinki.fi

Asstract — The lichen Cladonia corymbescens (Cladoniaceae, Ascomycota) has been

reported to have two main ranges, one in Melanesia and Australasia and another in the

Himalayas and surrounding areas. When specimens from New Caledonia, Bhutan, and

Thailand were subjected to molecular sequencing (ITS rDNA and RPB2), two distinct clades

were detected. The clades were also supported by morphological and geographical differences

supporting recognition of two distinct species. Cladonia corymbescens s. str. is present in

the Philippines, Melanesia, and Australasia, while the South East Asian mainland material is

referred to C. pseudofissa, a new combination at species level.

Key worps — Cladonia furcata group, lichens, molecular systematic, phylogeny, taxonomy

Introduction

One of the most common fruticose species of Cladonia in the mountains of

western Melanesia is Cladonia corymbescens. The nomenclature of this species

and its similarities with other species was thoroughly discussed by Stenroos

(1988), who also reported it from China and Nepal. However, Abbayes (1974)

was the first lichenologist to recognize this species from the Himalayas. His

opinion was also accepted by Awasthi & Ahti (2007) and Rai et al. (2014), who

stated that the species is widespread in the mountains in India and Nepal. In

his world monograph, Vainio (1887) could not give a definite opinion on the

type material of C. corymbescens from New Caledonia, and he referred the only

Himalayan specimen representing that species that he apparently examined to

C. erythrosperma var. thomsonii.

Several studies using molecular data have shown that many morphological

Cladonia species are not monophyletic (Kotelko & Piercey-Normore 2010,

Fontaine et al. 2011, Pino-Bodas et al. 2011, Steinova et al. 2013). The re-

examination of the morphology based on molecular data in lichenized fungi

has revealed that in several cases some phenotypical differences have been

overlooked (Argitiello et al. 2007, Nufez-Zapata et al. 2010, Parnmen et al.

92 ... Ahti, Pino-Bodas, & Stenroos

2013, Muggia et al. 2014). The aim of this study was to use DNA sequence

data to assess whether Cladonia corymbescens is a genetically homogeneous

species and to revise its classification based on the phylogenetic results and

morphological differences.

Materials & methods

Material for DNA analyses

Altogether 39 specimens of 29 species of Cladonia representing the supergroup

Cladonia in the provisional classification of Stenroos et al. (2002) were selected for a

phylogenetic analysis in this study. These specimens are preserved in the herbaria of

University of Helsinki (H), University of Turku (TUR) or Complutense University,

Biology Faculty (MACB); details of the collections are shown in TaBLE 1. Four specimens

of Cladonia corymbescens, two from New Caledonia and two from the Himalayas, were

available for DNA analyses, while 60 sequences had been generated in previous studies

(Stenroos et al. 2002; Pino-Bodas et al. 2010a, 2010b, 2012a, 2012b, 2012c, 2013a,

2013b). Cladonia rangiformis Hoffm. was selected as outgroup based on previous results

(Stenroos et al. 2002).

DNA extraction, PCR, and sequencing

Total DNA was extracted using DNeasy Plant Mini Kit (QUIAGEN, Germany)

following the manufacturer's instructions. In this study two genetic regions were

chosen: internal transcribed spacer of ribosomal DNA (ITS rDNA) and the second

largest subunit of RNA polymerase IT (RPB2). Although the ITS rDNA region has

been chosen as standard barcoding for fungi (Schoch et al. 2012), in Cladonia the

ITS rDNA frequently fails in the identifications (Kelly et al. 2011; Pino-Bodas et al.

2013b) and RPB2 gives better results (Pino-Bodas et al. 2013b). The primers used to

amplify the nuclear ITS nrDNA were ITS1F (Gardes & Bruns 1993) and ITS4 (White

et al. 1990), and for RPB2 they were RPB2dRaq/RPB2rRaq (Pino-Bodas et al. 2010a)

or CLRPB25F/CLRPB27R (Yahr et al. 2006). The amplification programs are listed in

Pino-Bodas et al. (2013a). PCR was carried out with Ready-to-Go-PCR Beads (GE

Healthcare Life Sciences, UK). Amplifications were prepared for a 25 uL final volume.

PCR was performed using a Mastercycler ep Gradient S (Eppendorf, Westbury, NY).

PCR products were purified with ExoSAP-IT (USB Corporation, USA) and sequenced

in Macrogen Europe (Amsterdam, Netherlands).

Alignments and phylogenetic analyses

The forward and reverse strands were assembled and edited in Sequencher™ v. 4.1.4

(Gene Codes Corporation, Inc., Ann Arbor, Michigan, USA). BLAST searches were

used to confirm that the obtained sequences belonged to Cladonia. The sequences were

manually aligned in BIOEDIT (Hall 1999). Nine ambiguous positions were manually

delimited and removed from the ITS rDNA alignment. The RPB2 alignment did not

have ambiguous positions.

Every region was separately analyzed by Maximum Likelihood (ML), with 500

bootstrap replicates to assess the support of each node. The conflict between the loci

was tested according Lutzoni et al. (2004). This method considers the existence of

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 93

TABLE 1. List of Cladonia specimens used in the molecular study with voucher

specimen information and GenBank accession numbers.

(The new sequences and the new combination are in bold font.)

SPECIES COLLECTION CODE ITS rDNA RPB2

C. acuminata USA, Ahti 63278 (H) 1ACUMI JN621932 JN621965

C. apodocarpa USA, Harris 54250 (H) 1APODO KC526127 KC526068

C. borbonica New Caledonia, Dennetiére 109 (H) AT686 AF455214 KP732364

C. cariosa Spain, Burgaz (MACB 94208) 5CARI JN621909 JN621941

C. cartilaginea Brazil, Stenroos 4926 (TUR) LK45 AF455212 —

C. ceratophylla Brazil, Stenroos 5081 (TUR) LK37 AF455171 —

C. conista Spain, Burgaz (MACB 92796) 2HUMIL JF926613 JF926567

Russia, Zolotuchin 26a (H) 1CONIST JF926633 JF926568

C. corsicana Spain, Burgaz (MACB 100763) SP1 JE288797 JF288833

Spain, Burgaz (MACB 101074) SP2 JF288798 JF288834

C. corymbescens New Caledonia, Dennetiére 45 (TUR) AT680 AF455235 KP732366

New Caledonia, Christenhusz 6169d (H) CL166 KP732363 KP732367

C. cyathomorpha Spain, Burgaz (MACB) 1CYATH KC415941 KC525275

Norway, Ahti 68660 & Tonsberg (H) 3CYATH KC415943 = KC525276

C. firma Spain, Burgaz (MACB 91619) 1FIRM FM205907 FM207568

Spain, Burgaz (MACB 90655) 7FIRM FM205910 FM207576

C. foliacea Portugal, Burgaz (MACB 90506) 1FOL FM205894 FM207569

C. furcata USA, Ahti 58283 (TUR) AT638 AF455220 KP732369

C. humilis Spain, Burgaz (MACB 95931) 9HUMIL JF926615 JF926576

Croatia, Burgaz (MACB 101103) 20HUMIL JF926621 JF926580

C. multiformis Canada, Ahti 57065 (H) LK70 AF455213 KP732370

C. nana Brazil, Stenroos 4940 (TUR) LK34 AF455211 —

C. neozelandica New Zealand, Wirth 28180 (TUR) AT590 AF455206 —

C. petrophila USA, Ahti 56654 (H) LK68 AF455222. —

C. peziziformis USA, Stenroos 5198 (TUR) AT631 AF455221 —

USA, Ahti 56670 (H) LK38 AF455182 —_

C. pityrophylla Brazil, Stenroos 5096 (TUR) LK40 AF455238 —

C. pseudofissa Thailand, Parnmen 285 (H) 1CORYMB KP732362 KP732365

Bhutan, Sgchting 9206 (H) LK35 AF455239 KP732368

C. pulvinata Spain, Burgaz (MACB 91646) 4PUL FM205911 =FM207579

C. pulvinella USA, Ahti 69191 (H) CLCAL4 KC415963 KC525287

C. pyxidata Greenland, Hansen (H) 17PYXI KC415983 = KC525269

C. rangiformis Netherlands, Van der Goes et al. (H) 5RANG JN811400 JN811429

Sweden, Skytén (H) 6RANG JN811401 JN811430

C. scabriuscula Canada, Ahti 56969 (H) LK11 AF455217 KP732371

C. subcariosa USA, Moore (H) 1SUBCARI JN621936 JN621969

C. subconistea Korea, Moon 7188 (H) 1SUBCONI KC415949 KC525296

C. subulata Spain, Burgaz (MACB 93151) 1SUBU FN86566 HM243210

C. turgida Canada, Lendemer (H) 1TURG JF288801 KC526089

94 ... Ahti, Pino-Bodas, & Stenroos

incongruity whenever a clade was supported with a bootstrap of more than 75% in a

locus, while in other loci the individual sequences of this clade were part of another

clade with bootstrap support =75%. The trees were checked manually. No conflicts

were detected and the datasets were combined. The combined dataset was analyzed by

Maximum Parsimony (MP), ML, and Bayesian analyses. MP analysis employed PAUP

version 4.0.b.10 (Swofford 2002), using heuristic searches with 1000 random taxon-

addition replicates with TBR branch swapping and MulTrees option in effect, and equally

weighted characters and gaps treated as missing data. Bootstrap with 1000 replicates

was used as the confidence analysis, using the heuristic option. The ML and Bayesian

analyses were run considering four partitions (ITS rDNA and each codon position of

RPB2). Maximum Likelihood analyses (ML) were done in RAXML 7.04 (Stamatakis

2006), assuming the model GTRGAMMA. A fast bootstrap with 500 replicates was

implemented to assess the support of each node. Bayesian analysis was carried out using

the program MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001). The models of evolution

were selected with MrModeltest (Nylander 2004) under AIC criterion. The models

selected were GTR+I+G for ITS rDNA and SYM+1+G for RPB2. Posterior probabilities

(pp), approximated by sampling trees using Markov Chain Monte Carlo (MCMC), of

each branch were calculated by counting the frequency of trees that were visited during

the course of the MCMC analysis. Model parameters were estimated in each analysis for

5,000,000 generations sampled in four simultaneous chains, and every 1000th was saved

into a file. When the analysis was finished we checked that standard deviation between

the runs was <0.005. The program AWTY (Nylander et al. 2008) was used to determine

when the chains reached the stationary stage. The first 1000 trees were deleted as the

“burn in” and the 50% majority-rule consensus tree was calculated using the “sumt”

command of MrBayes.

Morphology and chemistry

About 63 herbarium specimens of Cladonia corymbescens s. lat. deposited in Helsinki

(H) were morphologically re-evaluated after the phylogenetic analyses. Authors Ahti

and Stenroos had previously studied many additional specimens in various herbaria

[e.g., Geneva (G), Leiden (L), Lucknow (LWG, LWG-LWU, LWG-AWAS)] as well as

material borrowed from Edinburgh (E); see also the specimen lists in Stenroos (1988)

and Rai et al. (2014).

The secondary metabolites were analyzed by thin layer chromatography (TLC)

according to standardized procedures (Orange et al. 2001) using the solvents A (Toluene:

dioxane: acetic acid, MERCK) and B (Hexane: diethylether: formic acid, MERCK).

Results

In this study we generated 10 new sequences (2 of ITS rDNA and 8 of RPB2).

The combined dataset contained 1332 unambiguous characters (597 of ITS

rDNA and 735 of RPB2), 267 of which were parsimony informative (152 of ITS

rDNA and 115 of RPB2). The MP analysis generated two equally parsimonious

trees of 886 steps. The ML analyses of the combined dataset generated a tree

with likelihood value of -LnL = 6355.82, while the mean of likelihood of the

Bayesian analysis was —LnL = 6589.51.

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 95

77 --C. apodocarpa 1APODO

C. petrophila LK68

C. borbonica AT686

100"— ©. corsicana SP 1

C. corsicana SP2

1por— C. cyathomorpha 1CYATH

C. cyathomorpha 3CYATH

C. conista 1CONIST

C. conista 2HUMIL

C. subconistea 1SUBCONI

C. pulvinella CLCAL4

9gr— C. cartilaginea LK45

87 C. neozelandica AT590

85 C. nana LK34

100(— C. peziziformis AT631

C. peziziformis LK38

C. furcata AT638

C. multiformis LK70

100 C. humilis 2OHUMIL

C. humilis 9HUMIL

C. scabriuscula LK11

100[— ©. corymbescens AT680 (New Caledonia)

C. corymbescens CL166 (New Caledonia)

gi1-— C. ceratophylla LK37 A

C. pityrophylla LK40

1por— C. pseudofissa comb. nov. 1CORYMB (Thailand)

C. pseudofissa comb. nov. LK35 (Bhutan)

100/— ©. firma 1FIRM

C. firma 7FIRM

78[—- C. acuminata 1ACUMI

C. cariosa 5CARI

C. foliacea 1FOL

100

100 g9f— C. pulvinata 4PUL

95 C. subcariosa 1SUBCARI

le C. subulata 1SUBU

C. turgida 1TURG

C. pyxidata 17PYXI|

C. rangiformis 5RANG

C. rangiformis BRANG

Fic. 1. Cladonia spp. phylogeny: consensus tree of the two most parsimonious trees based on the

concatenated matrix of ITS rDNA and RPB2. The numbers on the branches represent bootstrap

values (>75%).

In all the analyses the specimens from the Himalayan region and the

specimens from New Caledonia formed two clades; however, the relationships

96 ... Ahti, Pino-Bodas, & Stenroos

om Lome C. porbonics RvO8S

¢. corsicana

74 re ose i fe CYATH

0.99/82] 1/100 . cyatnomorpna

C. cyathomorpha 3CYATH_

1/100 C. cartilaginea LK45

-_C, neozelandica AT590

C. nana LK34 .

C. peziziformis AT631

C. peziziformis LK38

C. furcata AT638

C. multiformis LK70

1/100 - C. humilis ZOHUMIL

C. humilis 9HUMIL

0.97/77 1/100 -C. conista 1CONIST

1/88 C. conista 2HUMIL

C. subconistea_1SUBCONI

C. pulvinella CLCAL4

C. scabriuscula

1/88 C. ceratophylla LK37

ane eee aes eS pityrophylla LK40

C. pseudofissa comb. nov. 1CORYMB (Thailand)

C. pseudofissa comb. nov. LK35 en

1/100; C. corymbescens AT680 (New Caledonia

C. corymbescens CL166 (New Caledonia)

al Se eC apodocarpa 1APODO

C. petrophila LK6

C. pulvinata 4PUL

C. subcariosa 1SUBCARI

C. subulata 1SUBU

- C. turgida 1TURG

C. acuminata 1ACUMI

C. cariosa 5CARI

14100 - C. rangiformis SRANG

C. rangiformis BRANG

Fic. 2. Cladonia spp. phylogeny: 50% consensus majority tree of Bayesian analysis based on the

concatenated matrix of ITS rDNA and RPB2. Posterior probability of bayesian analysis (>0.95) and

bootstrap values of ML analyses (>75%) are shown on the branches.

were not the same in all the analyses. In the MP analysis (Fie. 1) the specimens

from the Himalayan region (in the tree C. pseudofissa) are closely related with

C. pityrophylla Nyl. and C. ceratophylla (Sw.) Spreng. (Fic. 1, clade A), but this

relationship was not well supported (bootstrap <70%). The specimens from

New Caledonia cluster in a clade with specimens representing the C. furcata

group, C. humilis group, C. corsicana (Rondon & Vézda) Pino-Bodas et al.,

C. borbonica Nyl., C. cartilaginea Mill. Arg., and C. neozelandica Vain. (Fie. 1,

clade B). The ML and Bayesian analyses generated trees with the same topology,

and only the 50% consensus majority-rule tree from Bayesian analyses is

shown (Fic. 2). The specimens from New Caledonia (C. corymbescens) and the

specimens from the Himalayan region (C. pseudofissa) formed two clades that

are closely related, although the relationship was not supported in any of the

analyses.

Other differences between the MP tree and the Bayesian tree are that

C. pyxidata (L.) Hoffm. was shown as basal in the MP analysis, while the

Bayesian analysis showed C. cariosa (Ach.) Spreng. and C. acuminata (Ach.)

Norrl. in the basal clade. Additionally, the clade of C. conista (Nyl.) Robbins,

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 97

C. subconistea Asahina, and C. pulvinella S. Hammer is closely related to a clade

of C. cyathomorpha Stirt. ex Walt. Watson, C. corsicana, and C. borbonica in the

MP analysis, but not in the Bayesian analysis.

Taxonomy

Cladonia corymbescens Ny]. ex Leight., Ann. Mag. Nat. Hist., ser. 3, 18: 407.

1866. Fig. 3A

TyPeE (Abbayes 1974: 112): New Caledonia, Mont de M’bée, 1855-1860 E. Vieillard 1785

(PC, lectotype, G, H-NYL 38414, PC-Lenormand, isolectotypes). [The lectotypification

by Abbayes was overlooked by Ahti in Stenroos (1988: 126), Ahti (1993) and other

recent authors, but the type collection cited is the same].

= Cladonia novoguineensis Zahlbr., Bot. Jahrb. Syst. 62: 455. 1929.

Type: Papua New Guinea [“Nordéstliches Neu-Guinea, Kaiser Wilhelmsland”], Morobe

Prov., Sarawaged Mts. [“Saruwaged-Gebirg”], 3600-4000 m, C. Keysser 66 [“68”] (W,

holotype).

Primary thallus 0.5-1 x 1.5-2 mm, soon disappearing. Podetia (2—)3-7 cm tall,

0.7-1.5 mm thick, pale to dark brown, medulla hardly melanotic at base, erect,

dichotomously, trichotomously or irregularly richly branched, axils open, at

least some podetia stout, sterile podetia have thin, subulate tips, fertile podetia

becoming thicker, forming characteristic lateral fissures and perforations,

branches often flattened. Surface highly discontinuously corticated, especially

towards the base, in part sparsely microsquamulose or with larger squamules,

sometimes verruculose, apical parts hardly pruinose. Podetial wall 120-220 um

thick, with indistinct cortex 10-20 um, medulla 150-200 um, and stereome

50-60 um, hard. Pycnidia terminal or sometimes lateral on podetia, rarely

on basal squamules, containing hyaline slime, 1.5 mm in diam, conidia 6-7

x 1 um. Apothecia 0.8-1 mm wide, terminal, on every tip on fertile podetia,

sometimes deformed, brown, spores not observed.

CHEMISTRY—K+ yellow, PD+ red, rarely PD-. CHEMOTYPE 1: Atranorin

(major), fumarprotocetraric acid (major), protocetraric acid (minor),

confumarprotocetraric acid (minor), rarely rangiformic and norrangiformic

acids, occasionally 1-4 unknown minor substances and 1-3 terpenoids.

CHEMOTYPE 2: Atranorin only, the fumarprotocetraric acid complex lacking

(then PD-) (Stenroos 1988). If chemotype 2 proves to represent a distinct taxon,

the name Cladonia novoguineensis is applicable for that (no fresh material for

DNA analysis was available).

HasitatT— Terrestrial in bogs, grasslands, road banks, rarely on tree stumps,

at 1500-4400 m in Papua New Guinea (Stenroos 1988: 126).

DISTRIBUTION—Australia (New South Wales, Queensland), Indonesia

(Java), New Caledonia, New Zealand (North Island), Papua New Guinea,

Philippines (Luzon).

98 ... Ahti, Pino-Bodas, & Stenroos

ADDITIONAL SPECIMENS EXAMINED (See also Stenroos 1988) — NEW CALEDONIA,

between Yaté and Goro, 1966, Hill 11741 (BM, H). PAPUA NEW GUINEA, EAsTERN

HIGHLANDS, Mt. Wilhelm, Pindaunde Lake, 3540 m, 1965, A. C. Jermy 5327 (BM, H;

richly fertile!)}; MOROBE PRov., Mt. Sarawaket Southern Range, 4 km NEE of Lake

Gwam, 2850 m, 1981, T. Koponen 31662 (H, LAE); SOUTHERN HIGHLANDS, Mt. Giluwe,

3500 m, 1982, H. Streimann 24151 (CANB, H, LAE; chemotype 2). PHILIPPINES,

Luzon, Mountain Prov., Alab on road Baguio to Bontoc, 850 m, 1976, W. S. Gruezo

2030 (CAHP, H).

Cladonia pseudofissa (Asahina) Ahti, Pino-Bodas & S. Stenroos, stat. et

comb. nov. Fic. 3B

MycoBank MB 809612

= Cladonia rangiformis var. pseudofissa Asahina, Fl. E. Himalaya: 595. 1966.

TyPE: India, Sikkim, Jongri, 4000 m, 1960, M. Togashi 157 (TNS, holotype).

= Cladonia erythrosperma var. thomsonii Vain., Acta Soc. Fauna FI.

Fenn. 4: 376. 1887, syn. nov. [non Cladonia thomsonii Ahti].

Type: India, "India orientalis’, [1847-51], J. D. Hooker & T. Thomson 2145 (PC,

holotype).

= Cladonia rangiformis var. incurva Mull. Arg., Flora 74: 72.

1891, syn. nov. [non Cladonia incurva Ahti].

Type: India, Himachal Pradesh (?), Damodar [“Damdar”] Valley, 4200 m, J. F Duthie

(BM, holotype).

The earliest names for C. pseudofissa are C. erythrosperma var. thomsonii and C.

rangiformis var. incurva, but they cannot be adopted at species level because these

epithets are already occupied in Cladonia by C. thomsonii Ahti (1978) and C. incurva

Ahti (1961).

Primary thallus rarely visible, soon disappearing, consisting of minute

squamules. Podetia 1-4(-5) cm tall, to 1 mm thick, almost white, ashy grey

or more rarely dark brown, medulla clearly melanotic at base; often very

delicate, ascendant to erect, usually somewhat curly in appearance, without any

stouter sterile podetia, repeatedly subcorymbosely dichotomously branched,

apices subulate, ascyphose, axils usually perforated. Fertile podetia somewhat

thicker. Surface very matt, minutely fibrous or pruinose, often very smooth and

almost continuously corticate, with a few very small squamules, longitudinally

somewhat fissured, esorediate. Podetial wall 150-160 um thick, with distinct

cortex 10-15 um, medulla 75-100, stereome 25-50 um thick, softish rather

than cartilaginous. Pycnidia always at tips of podetia, shortly stalked, pyriform,

black, c. 2 x 1 mm; conidia or colour of slime not observed. Apothecia dark

brown, 0.5-1 mm, subglobose, on tips of flattened branchlets, spores not

observed.

CHEMISTRY—K+ yellow, PD+ orange red. Contains atranorin and the

fumarprotocetraric acid complex.

HasitatT—On ground, often on thin soil over rocks.

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 99

Fic. 3. A: Cladonia corymbescens (Papua New Guinea, T. Koponen 31662, H).

B: Cladonia pseudofissa (Nepal, P. Ozenda 147, H). Scale bar = 1 cm.

DISTRIBUTION—Bhutan, China (Sichuan, Tibet, Yunnan), India (Arunachal

Pradesh, Himachal Pradesh, Kerala, Sikkim, Tamil Nadu, Uttarkhand, West

Bengal), Malaysia (Sabah), Nepal, Thailand.

ADDITIONAL SPECIMENS EXAMINED — BHUTAN, Paro District: below Jaley la

Dzong, 2500-3000 m, 1998, U. Sochting 9206 (C, H; DNA voucher, Stenroos et al. (2002)

100 ... Ahti, Pino-Bodas, & Stenroos

and GenBank, as C. corymbescens). CHINA, SICHUAN: Miyi Co., 3200 m, 1983, L. S.

Wang 83-834 (H, KUN). TrpeT: Kang Ding Co., 1976, Z. Mu 5304 (H, KUN); YUNNAN:

Lijiang Co., 1985, L. S$. Wang 85-140 (H, KUN). INDIA, Srxxim: North Sikkim

District, Phune-Yakche, 3000 m, 1996, G. P. Sinha 1103 (ASSAM, H). UTTARAKHAND:

Pithoragarh District, Satgarh-Dhawj, 2700 m, 1989, D. K. Upreti L-18438 (H, LWG).

[For details of further Indian collections, see Rai et al. 2014, as C. corymbescens.] NEPAL,

FARWESTERN REGION: Seti, Dadachaur, Nhuna Khola, 2700 m, 1973, P. Ozenda 147C

(Gy Eh).

Discussion

Cladonia pseudofissa was included by Abbayes (1974) in C. corymbescens.

He also thought that C. perfossa Nuno, described from Taiwan, belongs

here. However, as discussed by Ahti & Lai (1979) and Stenroos (1988: 126),

C. perfossa is probably a distinct species (podetia short and thick; a topotype

re-examined in H), although still very poorly known.

Asahina (1966) clearly included the present species in C. rangiformis because

he found that it contains atranorin in addition to fumarprotocetraric acid;

such a strain is known in C. rangiformis. In habit C. pseudofissa does resemble

C. rangiformis to some extent, but C. rangiformis is not known to range east of

northwestern Iran (Litterski & Ahti 2004: 233). They are distinguished by the

longitudinally split podetia and melanotic part at the base of C. pseudofissa.

The two segregated species generally differ in habit and colour,

C. corymbescens being taller, erect, brownish (much resembling C. furcata

(Huds.) Schrad.) while C. pseudofissa is smaller, thinner, not readily erect,

and with an ashy grey or whitish tint. In addition, C. corymbescens is hardly

blackening (melanotic) at base, whereas C. pseudofissa is clearly blackening.

The phylogenetic analyses by Stenroos et al. (2002) demonstrated that

C. corymbescens was not closely related to the species of the C. furcata group

(C. farinacea (Vain.) A. Evans, C. furcata, C. multiformis G. Merr., and

C. scabriuscula (Delise) Nyl.); it seemed to be related to C. petrophila R.C.

Harris and C. apodocarpa Robbins instead. The phylogenetic study of Cladonia

in Thailand by Parnmen et al. (2008) grouped one specimen corresponding to

C. pseudofissa with C. rudis Ahti & Parnmen. Morphologically this species

is closely related to the species of C. furcata (Ahti et al. 2008). Our present

phylogenetic analyses do not allow conclusions on the phylogenetic

relationships of C. corymbescens and C. pseudofissa. Until further Cladonia

species are included in the phylogenetic analyses, the placement of these species

will remain uncertain. The ML and Bayesian analyses suggest that they might be

related, but this relationship is not supported, and the branches in both analyses

are very long. The MP analysis groups Cladonia pseudofissa with C. ceratophylla

and C. pityrophylla, but this relationship is not well supported either. A denser

sampling of taxa is needed to clarify the phylogenetic relationships between

the species.

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 101

As far as is known, the geographic ranges of C. corymbescens and C. pseudofissa

do not overlap; C. corymbescens is primarily Australasian to Melanesian and

C. pseudofissa is Himalayan in distribution. However, the Cladonia flora of

Indonesia is very poorly known.

Various data support C. pseudofissa asa species distinct from C. corymbescens.

The phylogenetical analyses based on two independent loci separate Cladonia

pseudofissa and C. corymbescens into two well-supported clades, clades that

are also supported in the single gene analyses (data not shown). According to

the genealogical concordance in phylogenetic species recognition (Taylor et al.

2000), these clades represent different species. In addition, morphological and

distributional differences confirm that they are two different taxa.

Acknowledgments

We are grateful for fresh material of Cladonia collected by Dr. Maarten Christenhusz

(London) in New Caledonia. We are also grateful to Dr. Philippe Clerc (Geneva), who

sent our herbarium (H) a large collection of Cladonia from the Himalayas. Dr. Jaana

Haapala helped us in photographing. Our sincere thanks to Dr. A. R. Burgaz and

Dr. M. Piercey-Normore for their valuable comments to improve the manuscript.

Literature cited

Abbayes H des. 1974. Cladonia du Népal. Lichenes, Cladoniaceae (Flechten des Himalaya 9).

Khumbu Himal 6(2): 111-116. Innsbruck, Miinchen: Universitatsverlag Wagner.

Ahti T. 1961. Taxonomic studies on reindeer lichens (Cladonia, subgenus Cladina). Ann. Bot. Soc.

Zool. Bot. Fenn. “Vanamo’ 32(1): 1-160.

Ahti T. 1978. Two new species of Cladonia from western North America. Bryologist 81: 334-338.

Ahti T. 1993. Names in current use in the Cladoniaceae (lichen-forming ascomycetes) in the ranks

of genus to variety. Regnum Vegetabile 78: 59-106.

Ahti T, Lai MJ. 1979. The lichen genera Cladonia, Cladina and Cladia in Taiwan. Ann. Bot. Fenn.

16: 228-236.

Ahti T, Parnmen S, Mongkolsuk P. 2008. Three new species of Cladonia from Thailand. Sauteria

i Regal Ese be

Argitello A, Del Prado R, Cubas P, Crespo A. 2007. Parmelia quercina (Parmeliaceae, Lecanorales)

includes four phylogenetically supported morphospecies. Biol. J. Linn. Soc. 91: 455-467.

http://dx.doi.org/10.1111/j.1095-8312.2007.00810.x

Asahina Y. 1966. Lichens. 592-610, in: H Hara (ed.). The flora of Eastern Himalaya. University of

Tokyo.

Awasthi DD, Ahti T. 2007. Cladonia P. Browne (Cladoniaceae). 79-116, in: DD Awasthi (ed.).

A compendium of the macrolichens from India, Nepal and Sri Lanka. Dehra Dun: Bishen

Singh Mahendra Pal Singh.

Fontaine KM, Ahti T, Piercey-Normore MD. 2010. Convergent evolution in Cladonia gracilis and

allies. Lichenologist 42: 323-338. http://dx.doi.org/10.1017/S0024282909990569

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for Basidiomycetes.

Application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucleic Acids Symp. Ser. 41: 95-98.

102 ... Ahti, Pino-Bodas, & Stenroos

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogenetic trees.

Bioinformatics 17: 754-755. http://dx.doi.org/10.1093/bioinformatics/17.8.754

Kelly LJ, Hollingsworth PM, Coppins BJ. 2011. DNA barcoding of lichenized fungi demonstrated

high identification success in a floristic context. New Phytol. 191: 288-300.

http://dx.doi.org/10.1111/j.1469-8137.2011.03677.x

Kotelko R, Piercey-Normore MD. 2010. Cladonia pyxidata and C. pocillum; genetic evidence to

regard them as conspecific. Mycologia 102: 534-545. http://dx.doi.org/10.3852/09-030

Litterski B, Ahti T. 2004. World distribution of selected European Cladonia species. Symb. Bot.

Ups. 34(1): 205-236.

Lutzoni E, Kauff F, Cox C, McLaughlin D, Celio G, Dentinger B, Padamsee M, Hibbett D, James

TY, Baloch E, Grube M, Reeb V, Hofstetter V, Schoch C, Arnold AE, Miadlikowska J, Spatafora

J, Johnson D, Hambleton S, Crockett M, Shoemaker R, Sung G-H, Liicking R, Lumbsch T,

O’Donnell K, Binder M, Diederich P, Ertz D, Gueidan C, Hansen K, Harris RC, Hosaka K,

Lim Y-W, Matheny B, Nishida H, Pfister D, Rogers J, Rossman A, Schmitt I, Sipman H, Stone J,

Sugiyama J, Yahr R, Vilgalys R. 2004. Assembling the fungal tree of life: progress, classification,

and evolution of subcellular traits. Am. J. Bot. 91: 1446-1480.

http://dx.doi.org/10.3732/ajb.91.10.1446

Muggia L, Pérez-Ortega S, Fryday A, Spribille T, Grube M. 2014. Global assessment of genetic

variation and phenotypic plasticity in the lichen-forming species Tephromela atra. Fungal

Diversity 64: 233-251. http://dx.doi.org/10.1007/s13225-013-0271-4

Nufez-Zapata J, Divakar PK, Del-Prado R, Cubas P, Hawksworth DL, Crespo A. 2010. Conundrums

in species concepts: the discovery of a new cryptic species segregated from Parmelina tiliacea

(Ascomycota: Parmeliaceae). Lichenologist 43: 603-616.

http://dx.doi.org/10.1017/S002428291100051X

Nylander JAA. 2004. MrModelTest 2.1. Program distributed by the author. Uppsala: Evolutionary

Biology Centre, Uppsala University.

Nylander JAA, Wilgenbusch DL, Warren, JC, Swofford DL. 2008. AWTY (Are we there yet?):

A system for graphical exploration of MCMC convergence in Bayesian phylogenetics.

Bioinformatics 24: 581-583. http://dx.doi.org/10.1093/bioinformatics/btm388

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London.

Parnmen S, Rangsiruji A, Mongkolsuk P, Ahti T. 2008. Phylogenetics of lichens in the genus

Cladonia (Cladoniaceae) in northern and northeastern Thailand. Sauteria 15: 385-402.

Parnmen S, Leavitt SD, Rangsiruji A, Lumbsch HT. 2013. Identification of species in the Cladia

aggregata group using DNA barcoding (Ascomycota: Lecanorales). Phytotaxa 115: 1-14.

http://dx.doi.org/10.11646/phytotaxa.115.1.1

Pino-Bodas R, Burgaz AR, Martin MP. 2010a. Elucidating the taxonomic rank of Cladonia subulata

versus C. rei (Cladoniaceae). Mycotaxon 113: 311-326. http://dx.doi.org/10.5248/113.311

Pino-Bodas R, Martin MP, Burgaz AR. 2010b. Insight into the Cladonia convoluta-C. foliacea

(Cladoniaceae, Ascomycota) complex and related species, revealed through morphological,

biochemical and phylogenetic analyses. Syst. Biodivers. 8: 575-586.

http://dx.doi.org/10.1080/14772000.2010.532834

Pino-Bodas R, Burgaz AR, Martin MP, Lumbsch HT. 2011. Phenotypical plasticity and homoplasy

complicate species delimitation in the Cladonia gracilis group (Cladoniaceae, Ascomycota).

Org. Divers. Evol. 11: 343-355. http://dx.doi.org/10.1007/s13127-011-0062-2

Pino-Bodas R, Burgaz AR, Martin MP, Lumbsch HT. 2012a. Species delimitations in

the Cladonia cariosa group (Cladoniaceae, Ascomycota). Lichenologist 44: 121-135.

http://dx.doi.org/10.1017/S002428291100065X

Cladonia pseudofissa comb. nov. (Southeast Asia) ... 103

Pino-Bodas R, Ahti T, Stenroos S, Martin MP, Burgaz AR. 2012b. Cladonia conista and C. humilis

(Cladoniaceae) are different species. Bibl. Lichenol. 108: 161-176.

http://dx.doi.org/1436-1698/2012/108-161

Pino-Bodas R, Martin MP, Burgaz AR. 2012c. Cladonia subturgida and C. iberica (Cladoniaceae)

form a single, morphologically and chemically polymorphic species. Mycol. Prog. 11: 269-278.

http://dx.doi.org/10.1007/s11557-011-0746-1

Pino-Bodas R, Ahti T, Stenroos S, Martin MP, Burgaz AR. 2013a. Multilocus approach to species

recognition in the Cladonia humilis complex (Cladoniaceae, Ascomycota). Am. J. Bot. 100:

664-678. http://dx.doi.org/10.3732/ajb.1200162

Pino-Bodas R, Martin MP, Burgaz AR, Lumbsch HT. 2013b. Species delimitation in Cladonia

(Ascomycota): a challenge to the DNA barcoding philosophy. Mol. Ecol. Resour. 13: 1058-1068.

http://dx.doi.org/10.1111/1755-0998.12086

Rai H, Khare R, Upreti DK, Ahti T. 2014. Terricolous lichens of India: taxonomic keys and

description. 17-294, in: H Rai, DK Upreti (eds). Terricolous lichens in India 2. Springer,

Heidelberg. http://dx.doi.org/10.1007/978-1-4939-0360-3_2

Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque A, Chen V, Fungal Barcoding

Consortium. 2012. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal

DNA barcode for Fungi. Proc. Natl. Acad. Sci. USA 109: 6241-6246.

http://dx.doi.org/10.1073/ pnas.1117018109.

Stamatakis A. 2006. RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

http://dx.doi.org/10.1093/bioinformatics/btl446

Steinova J, Stenroos S, Grube M, Skaloud P. 2013. Genetic diversity and species delimitation of

the zeorin-containing red-fruited Cladonia species (lichenized Ascomycota) assessed with ITS

rDNA and £-tubulin data. Lichenologist 45: 665-684.

http://dx.doi.org/10.1017/S0024282913000297

Stenroos S. 1988. The family Cladoniaceae in Melanesia. 3. Cladonia sections Helopodium, Perviae

and Cladonia. Ann. Bot. Fenn. 25: 117-148.

Stenroos S, Hyvonen J, Myllys, L, Thell, A, Ahti T. 2002. Phylogeny of the genus Cladonia s. lat.

(Cladoniaceae, Ascomycetes) inferred from molecular, morphological, and chemical data.

Cladistics 18: 237-278. http://dx.doi.org/10.1006/clad.2002.0202

Swofford DL. 2002. PAUP*: phylogenetic analysis using parsimony (*and other methods), version

4.0b10. Sinauer, Sunderland, Massachusetts, USA.

Taylor JW, Jacobson DJ, Kroken S, Kasuga T, Geiser DM, Hibbett DS, Fisher MC. 2000.

Species recognition and species concepts in fungi. Fungal Genet. Biol. 29: 21-32.

http://dx.doi.org/10.1006/fgbi.2000.1228

Vainio EA. 1887. Monographia Cladoniarum universalis 1. Acta Soc. Fauna et Flora Fenn. 4: 1-509.

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, Orlando FL.

Yahr R, Vilgalys R, DePriest PT. 2006. Geographic variation in algal partners of Cladonia subtenuis

(Cladoniaceae) highlights the dynamic nature of a lichen symbiosis. New Phytol. 171: 847-860.

http://dx.doi.org/10.1111/j.1469-8137.2006.01792.x

MY COTAXON

http://dx.doi.org/10.5248/130.105

Volume 130, pp. 105-130 January-March 2015

Morphology and phylogeny of four new Lactarius species from

Himalayan India

KANAD Das", ANNEMIEKE VERBEKEN’, & JORINDE NUYTINCK”?

"Botanical Survey of India, Cryptogamy Unit, Central National Herbarium,

Howrah 711103, W.B., India

?Ghent University, Department of Biology, Research Group Mycology,

K.L. Ledeganckstraat 35, BE 9000, Gent, Belgium

*Naturalis Biodiversity Center, Section National Herbarium of the Netherlands,

PO. Box 9517, 2300RA Leiden, The Netherlands

* CORRESPONDENCE TO: daskanadbsi@gmail.com

ABSTRACT —Four new species of Lactarius are described from Himalayan India. Lactarius

olivaceoglutinus, L. pyriodorus, and L. yumthangensis belong to L. subg. Piperites and

L. indochrysorrheus is closely related to some representatives of L. subg. Russularia. An ITS

based phylogeny confirms the phylogenetic placement of the four new species, although the

monophyly of neither Lactarius subg. Piperites nor L. subg. Russularia can be confirmed.

The ITS data also suggest that the Indian species are closely related to some European and

American species.

Key worps — ectomycorrhizal fungi, macrofungi, Russulaceae, Sikkim

Introduction

After segregation of the well-known ectomycorrhizal milkcaps into Lactarius

Pers. and Lactifluus (Pers.) Roussel (Buyck et al. 2008, 2010), Lactarius sensu

novo comprises three subgenera (Verbeken & Nuytinck 2013): L. subg. Piperites

(Fr. ex J. Kickx f.) Kauffman, L. subg. Russularia (Fr. ex Burl.) Kauffman, and

L. subg. Plinthogali (Burl.) Hesler & A.H. Sm. Lactarius subg. Piperites can

be characterized as follows: pileus sticky to slimy/glutinous (more rarely dry

and shiny), often hairy and/or zonate or with watery spots; stipe dry or sticky,

often scrobiculate (Heilmann-Clausen et al. 1998, Basso 1999). In contrast,

representatives of L. subg. Russularia typically have dry (rarely somewhat

sticky) caps and stipes and colours that are predominantly orange or various

tinges of brown (Heilmann-Clausen et al. 1998).

106 ... Das, Verbeken, & Nuytinck

Sikkim, a small (0.22% of the land surface) state in India, lies in Eastern

Himalaya, which is part of the Himalaya Hotspot, one of the 34 Global

Biodiversity Hotspots (www.biodiversityhotspots.org). Within a very small

geographical area it is substantially diverse in flora, fauna, and mycobiota, the

latter seriously underexplored. During mycological expeditions to different

temperate to subalpine areas of the North District of Sikkim in 2009 (KD &

AV), 2011 (KD), and 2012 (KD), we collected ectomycorrhizal (ECM) fungi,

with Lactarius s.l. one of the dominant ECM genera in Dombang, Shingba

Rhododendron Sanctuary, and Zema. Dombang is a subalpine coniferous to

mixed (coniferous & broadleaf) forest dominated by Picea spinulosa, Abies

densa, Tsuga dumosa, and Larix griffithii. Shingba Rhododendron Sanctuary

contains subalpine coniferous and mixed forests distributed in Yumthang

valley and its adjoining areas; apart from several Rhododendron species, this

protected area is dominated by Abies densa, Picea spinulosa, Tsuga dumosa,

Larix griffithii, Magnolia globosa, M. campbellii, Acer pectinatum, and Betula

utilis. Zema is a subalpine coniferous forest dominated by Abies densa.

Although Sikkim harbours a large number of russulaceous taxa, only 32

species (out of ca 210 taxa recorded from India) have been reported (Berkeley

1852; Das 2009; Das et al. 2010, 2013; Das & Verbeken 2011, 2012; Van de Putte

et al. 2012). We describe here three new species of Lactarius subg. Piperites

(L. olivaceoglutinus, L. pyriodorus, and L. yumthangensis) and one new species

of L. subg. Russularia (L. indochrysorrheus).

Materials & methods

Morphological study

Macromorphological characters were described from the fresh basidiomata in

daylight. Colour codes and terms follow the COLOUR IDENTIFICATION CHART OF

BRITISH FuNGus FLora (Henderson et al. 1969, here prefixed by “a:”) or the METHUEN

HANDBOOK OF CoLourR (Kornerup & Wanscher 1981, here prefixed by “b:”). Spore

print colour codes follow Kranzlin (2005, here prefixed by “c:”).

Micromorphological structures such as basidia, hymenial cystidia, pileipellis,

stipitipellis, etc. were observed from free-hand sections of dry samples mounted in

a mixture of 5% KOH, 30% Glycerol, Phloxine, and Cotton Blue using an Olympus

CX41 compound microscope. Spores and spore ornamentation were studied in Melzer’s

reagent. Drawings were made from SEM images obtained in different magnifications.

Spore measurements were calculated based on 20 basidiospores per specimen;

dimensions represent minimum-mean-maximum length x minimum-mean-

maximum width, and Q = length/width ratio. Herbarium names are after Holmgren et

al. (1990).

DNA extraction, PCR amplification and sequencing

DNA was extracted from dried fruiting bodies according to Nuytinck & Verbeken

(2003) with slight modifications (Van de Putte et al. 2010). The internal transcribed

Lactarius spp. nov. (India) ... 107

TABLE 1. Collections of Lactarius and allied species used for molecular analyses.

SPECIES

Lactarius akahatsu

L. albocarneus

alboscrobiculatus

aspideus

atroviridis

auriolla

indochrysorrheus

azonites

Reis ia fot Enea

brunneoviolaceus

caespitosus

camphoratus

chichuensis

Ea itech

chrysorrheus

L. aff. chrysorrheus

L. citriolens

L. controversus

L. crassiusculus

L. cyathuliformis

L. flexuosus

L. formosus

L. fuliginosus

L. helvus

L. lilacinus

L. luridus

L. montoyae

L. necator

L. olivaceoglutinus

L. pallescens

L. peckii

L. pseudouvidus

VOUCHER

Verbeken 04-141

Verbeken 98-071

Verbeken 98-080

Le 175

Walleyn 3815

Verbeken 05-306

Walleyn 1601

KD 11-002 (Holotypus)

Verbeken 00-124

Verbeken 04-220

Verbeken 04-249

Walleyn 1605

Eberhardt 24.08. 04-8

Oberwinkler 46773

Wang 1236

Nuytinck 01-089

D’Hooge 08-020

Eberhardt 04.10. 02-8

Verbeken 04-212

Verbeken 05-359

Eberhardt 20.09.04-3

Verbeken 00-117

Le 369

Eberhardt 04.09.04-3

Walleyn 2136

Walleyn 3178

Eberhardt 06.09.02-1

Le 382 (Holotypus)

Basso 97-24

Eberhardt 08.09. 04-1

Walleyn 3774

Taylor 2003066

Eberhardt 17.09.04-3

Eberhardt 10.10. 04-4

Berteloot 11-011

Berteloot 11-012

Taylor 2004254

Walleyn 1455

KD 1065 (Holotypus)

Verbeken 04-231

KD 11-103 (Holotypus)

Nuytinck 04-020

Eberhardt 24.08. 04-14

Eberhardt 24.08. 04-13

HERBARIUM

GENT

CMU, SFSU, GENT

GENT

BSHC, GENT

GENT

UPS

TUB

HKAS

GENT

UPS

GENT

UPS

GENT

CMU, SFSU

UPS

GENT

URS

CMU, SFSU, GENT

priv. herb.

UPS

GENT

UPS

GENT

UPS

GENT

BSD

GENT

BSHC, GENT

GENT

UPS

ORIGIN

Thailand

France

Thailand

Czech Rep.

USA

Sweden

India

Belgium

France

Sweden

Germany

China

Belgium

France

Italy

USA

Sweden

Italy

Thailand

Sweden

Czech Rep.

Sweden

Thailand

Sweden

Belgium

Sweden

Belgium

Sweden

Belgium

India

France

India

USA

Sweden

GENBANK

KF133269

KJ742389

KF241545

EF141538

KJ742390

KF133270

KF133257

KJ742391

KF241540

KJ742392

KJ742393

KJ742394

KJ742395

FJ845421

AY606945

KF475766

KJ742396

KJ742397

KF133261

KJ742398

KJ742399

DQ422003

KF241544

EF560684

KF133266

KJ742400

KJ742401

DQ421992

EF141549

JQ446111

KF133263

KF133275

KJ742402

KJ742403

KJ742404

KF241547

KJ742405

KJ742406

KJ742407

EF560673

KF133276

KJ742408

DQ974747

KF133277

KJ742409

KJ742410

108 ... Das, Verbeken, & Nuytinck

L. pubescens Eberhardt 15.09.02-2 UPS Sweden DQ421996

L. pyriodorus KD 11-027 (Holotypus) | BSHC, GENT India KJ742411

L. quieticolor Eberhardt 10.09.04-1 UPS Sweden DQ422002

L. quietus Eberhardt 16.09.04-6 UPS Sweden KF133264

L. rufus Nuytinck 02-008 GENT Norway KF241543

L. sphagneti Walter 083 TUB Germany KJ742412

L. subdulcis Vervisch 06-024 GENT Belgium KF133279

L. subplinthogalus Verbeken 04-219 GENT USA KF241539

L. subsericatus Eberhardt 11.10.04-8 UPS Sweden DQ422011

L. thyinos Voitk 23-08-04 priv. herb. Canada KF133271

L. torminosus Walleyn 3183 GENT Czech Rep. KF133281

L. trivialis Van de Putte 10-011 GENT Russia KJ742413

Walleyn 3179 GENT Czech Rep. KJ742414

Eberhardt27.08. 02-17a UPS Sweden DQ421991

L. uvidus Van de Putte 10-027 GENT Russia KF241546

Nuytinck 01-033 GENT Finland KJ742415

Walleyn 1237 GENT France KJ742388

Walleyn 2119 GENT Sweden KJ742416

Eberhardt 28.08. 02-24 UPS Sweden KJ742417

L. vietus Eberhardt 11.10.04-1 UPS Sweden KF133267

L. vinaceorufescens Nuytinck 07-018 GENT Canada KF241542

L. yumthangensis KD 11-147 (Holotypus) | BSHC, GENT India KJ742418

Lactifluus volemus Eberhardt 09.08. 04-5 UPS Sweden DQ422008

Lactifluus vellereus Eberhardt 20.09.04-22 UPS Sweden DQ422034

Gace x Buyck 02-107 PC USA DQ421984

M. zonaria Desjardin 7442 SFSU, PC, BBH Thailand DQ421990

Russula cyanoxantha Eberhardt 29.09. 02-2 UPS France DQ422033

R. nigricans Eberhardt 20.09.04-7 UPS Sweden DQ422010

spacer region of the nuclear ribosomal DNA (ITS) was amplified and sequenced using

primers ITS1-F and ITS4 (White et al. 1990, Gardes & Bruns 1993). PCR amplification

protocols follow Le et al. (2007); sequencing was conducted with an ABI 3730XL or

ABI 3700 by MacroGEN (Amsterdam, The Netherlands). Sequences were assembled

and edited with Sequencher™ v4.9 (GeneCodes Corporation, Ann Arbor, Michigan,

US.A.). Specimens and sequences used in the phylogenetic analysis are shown in

TABLE l.

Alignment and phylogenetic analyses

Alignment was conducted with the on-line version of MAFFTv7 (Katoh & Standley

2013), applying the E-INS-I strategy, a very slow method recommended for fewer than

200 sequences with multiple conserved domains and long gaps. The alignment was

manually refined in BioEdit v7.0.9.0 (Hall 1999). Ambiguously aligned positions were

detected using Gblocks v0.91b (Castresana 2000), specifying less stringent conditions

than default. The minimum number of sequences for a conserved position and for a

flank position was set to half the number of sequences, the number of contiguous non-

conserved positions was set to 10, and the minimum length of a block after gap cleaning

to 5 and positions with gaps were not treated differently from other positions. The ITS

Lactarius spp. nov. (India) ... 109

L. pallescens DQ874747 USA

L. brunneoviolaceus AV04,220 FRA

L. brunneoviolaceus AV04,249 FRA

L. brunneoviolaceus RW1605 SWE

L. brunneoviolaceus UE24.08.04.8 SWE

se L. luridus AT2003066 SWE

L. luridus UE17.09.04.3 SWE

SrA |e. luridus UE10.10.04.4 SWE

L, luridus OB11.012 BEL

88 L. luridus OB11.011 BEL

L. luridus AT2004254 SWE

98] [IL iuridus RW1455 BEL

L. pyriodorus KD11.027 IND

L. aspideus RW3815 CZE

L. pseudouvidus UE24.08.04.14 SWE

egbL. pseudouvidus UE24.08.04.13 SWE

L. uvidus RW1237 FRA

L. wvidus RW2119 SWE

7 L. uvidus JNO1.033 FIN

92) L. uvidus UE28,08,02.24 SWE

L. wvidus KVP 10.027 RUS

L formosus LTH382 THA

L. olivaceoglutinus KD11.103 IND

L. caespitosus FJ845421

L. albocameus AV98.071 FRA

964 L. albocarneus AV98.080 FRA

L. alboscrobiculatus LTH175 THA

64 L, torminosus RW3183 CZE

L. pubescens UE15.09.02.2 SWE

L.controversus AV00.117 ITA

Ey L.quieticolor UE10.09.04.1 SWE

L. akahatsu JNO4.141 THA

84 L. thyinos AV23.08.04 USA

L. auriolla RW1601 SWE

0b L. citriolens UE20.09.04.3 SWE

L quietus UE 16.09.04.6 SWE

58 L. helws UE08.09.04.1 SWE

L. chichuensis XW1236 CHI

81 L. camphoratus FO46773 GER

L. cyathuliformis UE04.09.04.3 SWE

L, subdulcis JV06.024 BEL

3 L. subsericatus UE11.10.04.8 SWE

86 96

7 L. sphagneti LW083 GER

8 L. rufus JNO2,008 NOR

77 86) L. chrysonheus JNO1.089 BEL

100 fl. chrysorheus ED08.020 FRA

73 Lchrysortheus UE04.10.02.8 ITA

= 100 L. aff. chrysormheus AVO4.212 USA

L. aff. chrysomheus AVO5.359 USA

63|7—L. indochrysorrheus KD11.002 IND

64 L. vinaceorufescens JNO7.018 CAN

L. peckii JNO4.020 USA

L. yumthangensis KD11.147 IND

L, fexuosus RW2136 SWE

L flexuosus RW3178 CZE

SIL. flexuosus UE0S.09.02.1 SWE

L. trivalis UE27.08.02-17a SWE

L. trivalis KVP10.010 RUS

L. trivialis RW3179 CZE

100!

L lilacinus RW3774 BEL

L. atroviridis AVO5.306 USA

L. necator AV04.231 FRA

L. vetus UE11.10.04.1 SWE

100 ‘L. azonites AVO0.124 BEL

97) L. fuliginosus MTB97-24 SWE

L. subplinthogalus AV04,219 USA

77] 76 L. montoyae typus IND

L. crassiusculus LTH369 THA

100, 'M. ochricompacta BB02.107 USA

M. zonaria DED7442 THA

R. cyanoxantha UE29.09.02.2 SWE

cy R nigricans UE20.09.04.7 SWE

LE. volemus 09.08.04.5 SWE

% Lt vellereus UE20.09.04-22 SWE

“1

0.05

PLATE 1. The obtained ML topology based on ITS sequences of Lactarius, Multifurca, Russula, and

Lactifluus species. Bootstrap values >50% are indicated. Names in bold are the new Indian species

described in this paper. The scale bar represents the number of nucleotide changes per site.

sequences were partitioned into 5 partitions: the ribosomal genes 18S, 5.88, and LSU

and the spacer regions ITS1 and ITS2.

Maximum Likelihood (ML) analysis was performed in RAXML v7.0.3 (Stamatakis

2006), combining a ML search with the Rapid Bootstrapping algorithm for 1000

replicates. The model GTRGAMMA was estimated for each partition separately.

Results

Phylogeny

PLaTE 1 shows the obtained ML topology with bootstrap support (BS)

values >50% displayed. The tree shows a well-supported genus Lactarius (97%

BS). Within Lactarius, L. subg. Plinthogali receives a high support value (100%

BS), but monophyly of neither Lactarius subg. Piperites nor L. subg. Russularia

(sensu Heilmann-Clausen et al. 1998) is supported. Lactarius pyriodorus,

L. olivaceoglutinus, and L. yumthangensis are closely related to species

110 ... Das, Verbeken, & Nuytinck

traditionally placed in L. subg. Piperites. Lactarius indochrysorrheus is closely

related to the European L. chrysorrheus Fr. and the North American L. vinaceo-

rufescens A.H. Sm. and L. aff. chrysorrheus, which are assigned to L. subg.

Russularia in morphology-based taxonomy.

Species delimitation for this paper is mainly based on morphological

features. We were not able to include enough specimens/sequences from the

newly proposed species to test species delimitation in the phylogenetic tree.

Taxonomy

Lactarius indochrysorrheus K. Das & Verbeken, sp. nov. PLATES 2, 3, 6A,B,E

MycoBank MB804887

Differs from Lactarius chrysorrheus by its more viscid pileus with its pileipellis an

ixocutis covered by a thick gelatinous layer.

Type: India. Sikkim: North District, Dombang, 27°43’35.2”N 88°45’15.2”E, alt. 2920 m,

18.VIII.2011, K. Das, KD 11-002 (Holotype, BSHC; isotype, GENT).

EryMo_oey: an Indian look-alike of the European Lactarius chrysorrheus.

PiLEus 25-70 mm diam., convex with slightly pubescent and inrolled margin

when young, gradually planoconvex to applanate with slightly depressed centre,

rarely with a central papilla, sometimes becoming widely infundibuliform;

margin decurved with maturity, becoming very irregularly wavy; surface

smooth to greasy, viscid (sticky), shiny, brownish orange, pinkish buff to salmon

(a: 45) or orange (b: 6A6) (cinnamon (a: 10) to somewhat rusty after maturity),

gradually apricot cream to paler up to yellow (a: 5E), towards margin distinctly

zonate with several zones over the whole diameter but most dense in the center;

zones consisting of darker and watery spots; margin very faintly and shortly

striate. LAMELLAE subdecurrent, crowded (18-22/cm at pileus margin), pale

yellow (a: 4D) when young, gradually pink-spotted, finally becoming brown

to reddish brown, with lamellulae in 9 series; edge entire, concolorous. STIPE

45-65 x 6-8.5 mm, slender, subcylindrical to cylindrical or slightly widened

towards base; surface smooth, slightly greasy, strigose (hairy) at base, much

paler than the pileus, especially in the upper part (very pale pinkish), very pale

salmon to vinaceous, gradually darker up to rust to rusty tawny or pale brick

red. CONTEXT hollow in stipe, pinkish yellow (a: 4D) to pale salmon, turning

lemon yellow (a: 54) to greenish yellow (a: 57) after cut, changing to salmon

(a: 45) with FeSO, and greenish with Guaiac. LaTex abundant, white, turning

quickly to greenish yellow. Taste first mild, then becoming bitter and acrid.

Opour not distinctive. SPORE PRINT pale cream (c: 10 Y).

BASIDIOSPORES 6.4—7.4-8.5 x 5.6-6.2-6.9 um, (Q = 1.08-1.18-1.33),

subglobose to ellipsoid; ornamentation amyloid, <0.9 um high, composed

of conical to spine-shaped warts, with short or medium, irregular to regular

Lactarius spp. nov. (India) ... 111

PLATE 2: Lactarius indochrysorrheus (Holotype KD 11-002).

A. Fresh basidiomata showing lamellae and lamellulae. B. Basidiospores.

C. Marginal cells. D. Basidia. Scale bars: B = 5 um; C, D = 10 um.

112 ... Das, Verbeken, & Nuytinck

ridges which are aligned or connected and forming a partial to incomplete

reticulum; some isolated small warts present; plage sometimes distinct and

amyloid. Basrp1a 30-44 x 9-13 um, 4-spored, subclavate to ventricose;

sterigmata 2.5-4 x 1.5-2 um. PLEUROMACROCYSTIDIA 32-70 x 7.5-11 um,

fairly abundant, emergent <10-30 um, narrowly clavate, cylindric with tapered

apex or fusoid, often with mucronate to subcapitate apex, slightly thick-walled

(wall <0.5 um); content refractive. PLEUROPSEUDOCYSTIDIA filamentous, 3-5

um wide. LAMELLAR EDGE fertile with basidia, cystidia and marginal cells.

CHEILOMACROCYSTIDIA 35-55 x 8-9 um, moderately abundant, subclavate

to narrowly clavate or ventricose to fusoid sometimes with mucronate apex;

content refractive. CHEILOPSEUDOCYSTIDIA filamentous. MARGINAL CELLS

11-20 x 7-11 um, mostly clavate to subclavate, often multiseptate.

HYMENOPHORAL TRAMA with lactifers. PILEIPELLIS an ixocutis, <170 um thick,

with gelatinous layer mostly extended 10-15 um beyond the hyphal layer;

hyphae repent to suberect, <3.5 um wide, branched, septate. STIPITIPELLIS

an ixocutis, composed of repent hyphae mostly in parallel pattern; hyphae

<3.5 um wide, branched, septate. STIPE TRAMA mostly with numerous nested

sphaerocytes. CLAMP CONNECTIONS absent.

EcoLocy & DISTRIBUTION — Gregarious under Abies densa and Picea

spinulosa in subalpine coniferous or mixed (coniferous and broadleaf) forest.

August. Fairly common.

ADDITIONAL SPECIMENS EXAMINED: INDIA. SIKKIM: North District, Dombang,

27°43'35.2”N 88°45'15.2’E, alt. 2920 m, 14.VIII.2009, A. Verbeken, K. Das & K.V. Putte,

AV-KD-KVP 032 (BSHC, GENT); North District, Shingba Rhododendron sanctuary,

27°43'42.1”N 88°44'58.8”E, alt. 2889 m, 24.VIII.2011, K. Das, KD 11-082 (BSHC);

27°44’03.5”N 88°44’23.3’E, alt. 3208 m, 26.VIII.2012, K. Das, KD 11-107 (BSHC).

Notes — Zonate species with similar cap colours and milk that stains quickly

sulphur to greenish yellow are European L. chrysorrheus, North American

L. vinaceorufescens, and an undescribed American species [labeled L. aff.

chrysorrheus in the phylogenetic tree and TABLE 1]. ITS sequence comparison

suggests the new species described here is close to the three species but not

conspecific.

Lactarius chrysorrheus is less sticky, which is reflected in the pileipellis

structure, a loosely interwoven cutis without distinct slime layer (Heilmann-

Clausen et al. 1998). Lactarius vinaceorufescens differs by a pileipellis with a

relatively thin ixocutis and the lack of ixocutis in the stipitipellis (Hesler &

Smith 1979). Lactarius xanthogalus Verbeken & E. Horak, described from

Papua New Guinea (Verbeken & Horak 2000), has a zonate orange to paler cap

and latex that becomes immediately sulphur yellow but clearly differs by the

zebroid ornamented spores.

aks,

Lactarius spp. nov. (India) ...

Pleuromacrocystidia. B. Radial section through pileipellis.

PLATE 3. Lactarius indochrysorrheus (Holotype KD 11-002).

C. Pleuropseudocystidia. D. Cheilomacrocystidia. Scale bars = 10 um.

Pes

114 ... Das, Verbeken, & Nuytinck

Lactarius olivaceoglutinus K. Das & Verbeken, sp. nov. PLATES 4, 5, 6C,D,F

MycoBaAnk MB 804888

Differs from Lactarius albocarneus by its olivaceous pileus and its watery white latex that

turns faintly pinkish on bruised or cut gill tissue.

Type: India. Sikkim: North District, Shingba Rhododendron sanctuary, 27°44’19.5”N

88°44’25.9’E, alt. 3252 m, 26.VIII.2011, K. Das, KD 11-103 (Holotype, BSHC; isotype,

GENT).

EryMoLoey: named after the olive and extremely glutinous pileus of the basidiocarps.

Pireus 40-68(-120) mm diam., at first convex, gradually becoming

planoconvex with slightly depressed centre; surface smooth, viscid to highly

glutinous, olive yellow (b: 3D8) to olive brown (b: 4E8, 4F6) or golden brown

(b: 5D7) to greenish grey, grey-olive or darker towards centre and paler margin,

azonate; margin non-striate, incurved when young, decurved with maturity.

LAMELLAE adnexed to broadly adnate, crowded (17-18/cm at margin), with

lamellulae in 5-6 series, pale cream to cream; edge entire, concolorous. STIPE

40-75 x 12-17 mm, clavate to subclavate (broader towards base) or ventricose

(narrower towards apex and base); surface smooth or sometimes with shallow

depressions (but without scrobicules), sticky, pale salmon to pale yellow (b:

3A3 to 4A3), gradually whitish towards base; base strigose, pale cream to

white, sometimes with ochraceous spots. CONTEXT firm, stuffed to hollow in

stipe, cream (a: 4D) to pale cream, unchanging when exposed or with KOH,

changing to olivaceous (a: 62) to leaf green (a: 59) with Guaiac and pale green

with FeSO,. LATEX whitish, very watery, turning faintly pinkish on bruised or

cut lamellae. Taste mild and agreeable or slightly acrid. ODour indistinct.

SPORE PRINT pale yellow (c: 30Y, 2M).

BASIDIOSPORES 8.0-9.3-10.1(-11) x 6.7-7.3-7.8(-8.4) um, [Q = 1.16-1.28-

1.33(-1.44)], broadly ellipsoid to ellipsoid; ornamentation amyloid, <1.2 um

high, composed of narrow, low and high ridges aligned and parallel, forming

zebroid pattern, some connected with adjacent ones (but never forming a true

reticulum), with each high ridge bearing elongated to conical (with rounded

apex) warts; plage indistinct. Bastp1a 40-60 x 11.5-13 um, 2-4-spored,

ventricose; sterigmata 6-9 x 2-2.5 um. PLEUROMACROCYSTIDIA 57-88 xX

8-12 um, abundant, subcylindric or subclavate to fusiform with rounded,

mucronate to moniliform (rarely) apex, emergent <41 um; content dense.

PLEUROPSEUDOCYSTIDIA cylindrical to tortuous, rare, never emergent, 4-5

um wide; content refringent. LAMELLAR EDGE sterile with marginal cells

and some cystidia. CHEILOMACROCYSTIDIA 32-57 x 7-9 um, subcylindrical

to subfusiform, mostly with rounded apex. MARGINAL CELLS 13-23 x 6.5-9

um, narrowly clavate to clavate or subcapitate, slightly thick-walled, hyaline.

HYMENOPHORAL TRAMA with lactifers. PILEIPELLIS an ixotrichoderm, very

thick (often variable in thickness), 220-700 um thick, composed of erect,

Lactarius spp. nov. (India) ... 115

PLATE 4. Lactarius olivaceoglutinus (Holotype KD 11-103).

A. Fresh basidiomata showing lamellae and lamellulae. B. Basidiospores.

C. Basidia. D. Marginal cells. Scale bars: B = 5 um; C, D = 10 um.

116 ... Das, Verbeken, & Nuytinck

oe ST oy

103).

Pleuromacrocystidia. B. Radial section through pileipellis.

PLATE 5. Lactarius olivaceoglutinus (Holotype KD 11-

C. Cheilomacrocystidia. D. Pleuropseudocystidia. Scale bars = 10 um.

Lactarius spp. nov. (India) ... 117

PiateE 6. Lactarius indochrysorrheus. A. Fresh basidiomata. B. Latex oozing from cut lamellae.

E. Basidiospores (SEM). Lactarius olivaceoglutinus. C, D. Fresh basidiomata. FE. Basidiospores

(SEM). Scale bars: E, F = 5 um.

branched multi-septate hyphae (<3.5 um wide) and few lactifers embedded

in a slime layer of variable thickness underlying somewhat repent hyphae.

STIPITIPELLIS an ixotrichoderm, <120 um thick, composed of erect branched

118 ... Das, Verbeken, & Nuytinck

septate hyphae (<3 um wide) embedded in a slime layer. STIPE TRAMA with

numerous nested sphaerocytes. CLAMP CONNECTIONS absent.

EcoLoGy & DiIsTRIBUTION — Gregarious under Abies densa in subalpine

coniferous or mixed (coniferous and broadleaf) forest. August-September.

Fairly common.

ADDITIONAL SPECIMENS EXAMINED: INDIA. Sikkim: North District, Shingba

Rhododendron sanctuary, 27°45’11.1”N 88°43’47.5’E, alt. 3580 m, 29.VIII.2011, K.

Das, KD 11-144 (BSHC); North District, Dombang, 27°44’53.1”N 88°44’58.8”E, alt.

3058 m, 03.IX.2011, K. Das, KD 11-191 (BSHC); 27°44’51”N 88°46'32’E, alt. 2940 m,

15.VIII.2009, A. Verbeken, K. Das & K.V. Putte, AV-KD-KVP 09-049 (BSHC, GENT).

Notes — The combination of the viscid to glutinous pileus and stipe and the

ixotrichoderm pileipellis clearly places the present species in L. sect. Pallidini.

Lactarius olivaceoglutinus can be recognized by its distinctly glutinous grey-olive

to greenish grey pileus, pale salmon to pale yellow sticky stipe, whitish watery

latex that turns the bruised lamellae faintly pinkish, large spores with mostly

zebroid patterns of ridges (that carry warts), and its occurrence under Abies

sp. ITS sequences and morphology (mainly the thick pileipellis ixotrichoderm)

suggest a close relationship with the European L. albocarneus Britzelm.

[= L. glutinopallens FH. Moller & J.E. Lange] and North American L. caespitosus

Hesler & A.H. Sm. The pileus of L. albocarneus however, is dominantly cream

to at most olivaceous buff and its latex slowly turns to sulfur-yellow (Heilmann-

Clausen et al. 1998, Kranzlin 2005) and its spore ridges never bear elongated

to conical warts. Lactarius caespitosus also has a viscid cap with similar grey to

olive brown colours, but its spores are slightly larger with a distinctly low and

non-zebroid ornamentation (Hesler & Smith 1979).

In Sikkim L. elaioviscidus K. Das & Verbeken is another species with a

glutinous olivaceous pileus and watery white latex (Das & Verbeken 2011);

however, it is distinguished from L. olivaceoglutinus by the presence of a papilla,

its greyish yellow to yellowish white stipe without pinkish tinges, latex changing

to yellow-yellowish green after long exposure, and smaller basidiospores

without zebroid ornamentation (7.0-8.1-8.7 x 6.0-6.7-7.1 tm).

Lactarius pyriodorus K. Das & Verbeken, sp. nov. PLATE 7-9, 13A,B,F

MycoBank MB 804889

Differs from Lactarius formosus by lacking bundles of glutinous hairs on the pileus.

Type: India. Sikkim: North District, Dombang, 27°44’08.7”'N, 88°45’58.3”, alt. 2975 m,

19. VIII.2011, K. Das, KD 11-027 (Holotype, BSHC; isotype, GENT).

EryMo oy: in reference to the distinct pear-like odor of the fresh basidiocarps.

PILEus 35-77 mm diam., at first convex with slightly depressed centre, gradually

becoming planoconvex with depressed centre and finally broadly to widely

infundibuliform, rarely with a small, sometimes blunt central papilla; surface

Lactarius spp. nov. (India) ... 119

Pate 7. Lactarius pyriodorus (Holotype KD 11-027).

A. Fresh basidiomata showing lamellae and lamellulae. B. Basidiospores.

C. Basidia. D. Cheilomacrocystidia. Scale bars: B = 5 um; C, D = 10 um.

120 ... Das, Verbeken, & Nuytinck

smooth to finely rugulose under the slimy surface, viscid to highly glutinous

and slimy, mostly uniformly colored but sometimes with some indistinct

fading zones, bay (a: 19) to umber (a: 18), often sienna (a: 11) towards margin

with distinct rust (a: 13) to dark brick (a: 20) or brown vinaceous (a: 25), with

broader zonations towards centre and narrower zones towards margin; margin

non-striate to very finely and shortly striate, incurved when young, decurved

with maturity, becoming very irregularly wavy to undulate. LAMELLAE broadly

adnate to decurrent, crowded (15-16/cm at margin), sometimes forked near

the stipe, with lamellulae in 5 series, whitish cream to pale cream, turning livid

vinaceous (a: 77) to lilac (a: 79) or darker violet (b: 15D4-5, 15E5) after bruising;

edge entire, concolorous. STIPE 35-72 x 10-18 mm, cylindrical or broadest in

the middle, narrower towards base, often curved and locally swollen; surface

smooth, slightly viscid or sticky, not as viscid as pileus, dingy white to pale

cream, buff (a: 52) to vinaceous buff with maturity; base usually paler, strigose.

CONTEXT firm, rather thick in pileus center and thin in outer half, very soon

hollow in stipe, white to pale cream, changing lilac (a: 77; b: 13C3-4) after

exposure, leaf green with Guaiac, unchanging with FeSO, and KOH. LaTex

white to pale cream, watery, slowly staining the lamellae or context livid

vinaceous (a: 77) to lilac (a: 79), unchanging without the contact of lamellae

or context. TasTE mild at first, becoming slightly bitter and astringent. ODOUR

fruity, very sweet, like pears (as in Inocybe corydalina Quél.). SPORE PRINT pale

cream (c: 10Y).

BASIDIOSPORES 8.0-8.9-10.5 x 6.9-7.5-8.8 um, (Q = 1.11-1.18-1.32),

subglobose to broadly ellipsoid, rarely ellipsoid; ornamentation amyloid, <1.1

um high, composed of broad ridges and isolated warts forming a partial to an

incomplete reticulum; plage inamyloid or distally amyloid. Bastp1a 45-52 x

10.5-12 um, 4-spored, rarely 2-spored, clavate to ventricose; sterigmata 6-7

x 2-2.5 um. PLEUROMACROCYSTIDIA 55-80 Xx 7.5-11 um, abundant, fusiform

with mucronate to moniliform apex, emergent <33 um; content slightly dense.

PLEUROPSEUDOCYSTIDIA cylindric to slightly tortuous, never emergent, 3.5-4

um wide; content refringent. LAMELLAR edge sterile. CHEILOMACROCYSTIDIA

28-43 x 6.5-9 um, fusiform to lageniform, mostly with moniliform apex.

MARGINAL CELLS 15-35 x 7-10 um, cylindric to clavate, slightly thick-walled,

hyaline. SUBHYMENIUM $20 um thick. HYMENOPHORAL TRAMA with abundant

lactifers. PILEIPELLIS an ixocutis to ixotrichoderm, <320 um thick, composed

of few repent hyaline hyphae and abundant erect hyphae (2-4 um broad).

CLAMP CONNECTIONS absent.

EcoLocy & DISTRIBUTION — Gregarious to caespitose under Abies densa

and Tsuga dumosa in subalpine mixed (coniferous and broadleaf) forest.

August. Fairly common.

Lactarius spp. nov. (India) ... 121

ars.

10 um.

Pate 8. Lactarius pyriodorus (Holotype KD 11-027).

A. Pleuromacrocystidia. B. Marginal cells. Scale bars

122 ... Das, Verbeken, & Nuytinck

PLATE 9. Lactarius pyriodorus (Holotype KD 11-027):.

A. Radial section through pileipellis. Scale bar = 10 um.

Lactarius spp. nov. (India) ... 123

ADDITIONAL SPECIMENS EXAMINED: INDIA. SIKKIM: North District, Dombang,

27°44'08.7”'N 88°45’58.3’E, alt. 2975 m, 14. VIII.2009, A. Verbeken, K. Das & K.V. Putte,

AV-KD-KVP 050 & 051 (BSHC, GENT); alt. 2945 m, 23.VIII.2011, K. Das, KD 11-068

(BSHC); North District, Zema, 27°47’20.0’N 88°32’56.1”E, alt. 3079 m, 31.VIII.2012, K.

Das, KD 12-223, KD 12-226 (BSHC).

Notes — The viscid to glutinous subzonate pileus, ixocutis to ixotrichoderm

nature of the pileipellis, and latex that stains the context and lamellae distinctly

lilac to violet place this species in L. sect. Uvidi. Only one representative of

L. subg. Piperites with lilac milk has been described from Asia (northern

Thailand)—L. formosus H.T. Le & Verbeken (Le et al. 2007). Lactarius formosus

clearly differs from L. pyriodorus because its pileus is completely covered with

bundles of glutinous hairs.

In the ITS-generated phylogenetic tree, the resolution in the group of lilac

staining species is very low, and the Indian species is not distinctly separated

from a cluster with the European species Lactarius luridus (Pers.) Gray and

L. brunneoviolaceus M.P. Christ. We have previously observed that in this

group the morphological variation appears much higher than the ITS

molecular variation. Since we observe clear morphological differences

between L. pyriodorus and the European taxa (also not supported in this tree,

although generally accepted as separate species), we choose to present these

Indian specimens as a new species. Its darker colours and subzonate aspect

are shared with L. luridus, which typically associates with broadleaf trees and

can be separated by its pileus that is spotted but never with thick gluten (as

in L. pyriodorus). Also, in L. luridus, the stipe base is never strigose and the

distinctive sweetish pear-like odour is absent (Heilmann-Clausen et al. 1998).

Lactarius brunneoviolaceus and L. pseudouvidus Kihner are two arctic-alpine

species associated with Salix that differ greatly from our Indian species in their

very small habit. Lactarius brunneoviolaceus is further distinguished by its

cedar-oil odour and rather large spores (8.8-11.9 x 6.7-8.6 um) (Heilmann-

Clausen et al. 1998, Basso 1999). Lactarius pseudouvidus differs microscopically

by its distinctly lower (<0.3 um) spore ornamentation and differently shaped

hymenial cystidia (cylindric to subfusiform with rounded apex) (Heilmann-

Clausen et al. 1998). Lactarius pyriodorus is superficially similar to the European

L. uvidus (Fr.) Fr., which can also occur under coniferous trees, but it never has

such dark colours (Heilmann-Clausen et al. 1998), and its spores have rounded

warts and ridges forming a more incomplete reticulum.

This is for the first time that distinct sweetish odour reminiscent of pears

(also known from the European Inocybe corydalina) has been encountered in

Lactarius.

124 ... Das, Verbeken, & Nuytinck

Lactarius yumthangensis K. Das & Verbeken, sp. nov. PLATE 10-12, 13C,D,F

MycoBank MB 804890

Differs from Lactarius trivialis by its lighter spore print, smaller spores, and pileipellis

ixocutis.

Type: India. Sikkim: North District, Yumthang valley of Shingba Rhododendron

sanctuary, 27°46'51.9”N 88°42’37.6’E, alt. 3586 m, 30.VIIL.2011, K. Das, KD 11-147

(Holotype, BSHC, isotype, GENT).

EryMo_oey: after the name of the locality from where the type specimen was collected

PiLEus 37-98 mm diam., at first convex with inrolled margin, becoming

planoconvex to applanate with depressed indented centre or funnel shaped

at maturity, surface smooth, viscid when dry, slimy when moist, vinaceous

grey (a: 80) or slightly paler (purplish grey), mostly with distinct darker

spots forming zonations; margin non-striate, paler, decurved with maturity.

LAMELLAE broadly adnate to decurrent, close to medium crowded (9-11/cm

at margin), some forked, with lamellulae in 5 series, cream-yellow to yellow

(a: 6F) or ochraceous, sometimes with rusty spots after maturity, unchanging

when bruised; edge entire, concolorous. STIPE 21-50 x 9-22 mm, cylindrical to

ventricose, often constricted below the juncture of the lamellae, surface slightly

longitudinally venose, whitish on constriction, then vinaceous to vinaceous grey

(a: 80), cream yellow towards base, with ochraceous areas/spots towards base

after maturity. CONTEXT yellowish white to cream, hollow, multi-chambered

in stipe, becoming greenish with FeSO, and orange-yellow with KOH. LaTex

chalky white, changing to orange-yellow with KOH, turning pale cream (a: 3C)

after some time, slowly (on drying) greenish yellow (a: 57) after 2 hours (on cut

lamellae). TasTE acrid. ODour fruity. SPORE PRINT pale yellow (c: 30Y, 2M).

BASIDIOSPORES 6.4—7.1-8.0 x 5.3-6.0-6.9 um, (Q = 1.06-1.18-1.32),

subglobose to ellipsoid; ornamentation amyloid, <1 um high, composed of

rather regular, narrow and acute ridges mostly arranged in parallel groups

forming zebroid pattern, short ridges and isolated irregular warts present

between ridges, but, never forming reticulum; plage amyloid distally but

often not distinguishable. Bastp1a 38-58 x 9-11 um, 2-4-spored, clavate to

ventricose; sterigmata long, 5-9 x 2-2.5 um. PLEUROMACROCYSTIDIA 44-90

x 7.5-10 um, abundant, clavate to fusiform (sometimes with mucronate apex),

emergent <36 um; content slightly dense but never needle-like, slightly thick-

walled (wall up to 0.6 um). PLEUROPSEUDOCYSTIDIA irregularly tortuous

with rounded apex, 2.5-4 um wide; contents dense. LAMELLAR EDGE Sterile.

CHEILOMACROCYSTIDIA 27-55 x 8-9 um, subclavate or narrowly clavate to

fusoid, emergent <25 um, slightly thick walled (wall <0.7 um). MARGINAL

CELLS forming chains of elements, terminal cells 11-18 x 5-8 um, subcylindric

to subclavate or clavate, hyaline. SuBHYMENIUM <28 um thick, cellular.

Lactarius spp. nov. (India) ... 125

PLATE 10:. Lactarius yumthangensis (Holotype KD 11-147).

A. Fresh basidiomata showing lamellae and lamellulae. B. Basidiospores.

C. Pleuropseudocystidia. D. Marginal cells. Scale bars: B = 5 um; C, D = 10 um.

126 ... Das, Verbeken, & Nuytinck

Aan

Pea)

2% VV

PLATE 11. Lactarius yumthangensis (Holotype KD 11-147).

A. Pleuromacrocystidia. B. Cheilomacrocystidia. C. Basidia. Scale bars = 10 um.

Lactarius spp. nov. (India) ... 127

PLATE 12. Lactarius yumthangensis (Holotype KD 11-147).

A. Radial section through pileipellis. Scale bar = 10 um.

PILEIPELLIS an ixocutis, composed of narrow repent hyaline hyphae and broad

(<7 um) septate hyphae embedded in a layer of slime. CLAMP CONNECTIONS

absent.

EcoLoGy & DISTRIBUTION — Gregarious to caespitose under Betula utilis

in subalpine mixed (coniferous and broadleaf) forest. August-September.

Uncommon.

ADDITIONAL SPECIMENS EXAMINED: INDIA. Sikkim: North District, Yumthang

valley of Shingba Rhododendron sanctuary, 27°46’51.9”N 88°42’37.6’E, alt. 3586 m,

29.VIII.2011, K. Das, KD 11-150 (BSHC); 27°45’11.1”N 88°43’47.5’E, alt. 3580 m,

01.1X.2011, K. Das, KD 11-172 (BSHC).

Notes —The viscid to slimy zonate pileus and the ixocutis to ixotrichoderm

nature of the pileipellis undoubtedly place the present species in L. subg.

128 ... Das, Verbeken, & Nuytinck

ee 5 I

PLATE 13. Lactarius pyriodorus. A. Fresh basidiomata. B. Latex oozes out from cut lamellae.

E. Basidiospores (SEM). Lactarius yumthangensis. C. Fresh basidiomata. D. Basidiomata showing

constricted stipe-apex. FE Basidiospores (SEM). Scale bars: E = 5 um; F = 3 um.

Piperites. Lactarius yumthangensis can be recognized by its vinaceous grey to

purplish grey pileus, venose stipe with a constricted apex, chalky white latex

Lactarius spp. nov. (India) ... 129

that changes to orange-yellow with KOH and becomes greenish yellow after

long exposure, and growth under Betula sp.

Morphologically, L. flexuosus (Pers.) Gray and L. trivialis (Fr.) Fr. (both

reported from Europe) appear quite similar. However, both can be distinguished

from L. yumthangensis by the slightly darker spore print (c: 40Y, 5M). Moreover,

the white latex of L. flexuosus is unchanging (never becoming greenish yellow

after exposure), the stipe is not constricted at the apex (Kranzlin 2005), and

some cheilomacrocystidia have a moniliform apex (Heilmann-Clausen et al.

1998). In L. trivialis, spores are distinctly larger (7.3-10 x 5.9-7.8 um) and the

pileipellis is an ixocutis to ixotrichoderm (Heilmann-Clausen et al. 1998).

Acknowledgments

The authors are thankful to the Director, Botanical Survey of India, Kolkata (India)

and Department of Forest, Environment and Wild Life Management, Government

of Sikkim, for providing all kinds of facilities during this study. They are grateful to

Prof. N.S. Atri (India) and Dr. Xiang-Hua Wang (China) for reviewing the manuscript

and giving valuable suggestions. Thanks are also due to Ursula Eberhardt, who kindly

provided the sequences for the specimens. Assistance (during macrofungal surveys

to Dombang, Shingba Rhododendron sanctuary, and Zema) rendered by S.K. Rai,

S. Pradhan, R. Giri, R.K. Ram and P. Tamang (BSI, SHRC, Gangtok) is duly acknowledged.

Literature cited

Basso MT. 1999. Lactarius Pers. Fungi Europaei, vol. 7. Mycoflora, Alassio.

Berkeley MJ. 1852. Decades of Fungi: Decades XXXIX., XL. Sikkim and Khassya Fungi. Hooker’s

J. Bot. 4: 130-142.

Buyck B, Hofstetter V, Eberhardt U, Verbeken A, Kauff F. 2008. Walking the thin line between Russula

and Lactarius: the dilemma of Russula sect. Ochricompactae. Fungal Diversity 28: 15-40.

Buyck B, Hofstetter V, Verbeken A, Walleyn R. 2010. Proposal to conserve Lactarius nom. cons.

(Basidiomycota) with a conserved type. Taxon 59(1): 295-296.

Castresana J. 2000. Selection of conserved blocks from multiple alignments for their use in

phylogenetic analysis. Mol. Biol. Evol. 17(4): 540-552.

http://dx.doi.org/10.1093/oxfordjournals.molbev.a026334

Das K. 2009. Mushrooms of Sikkim 1: Barsey Rhododendron Sanctuary. Sikkim State Biodiversity

Board, Gangtok and Botanical Survey of India, Kolkata.

Das K, Verbeken A. 2011. Three new species of Lactarius from Sikkim, India. Cryptogamie

Mycologie 32(4): 365-381. http://dx.doi.org/10.7872/crym.v32.iss4.2011.365

Das K, Verbeken A. 2012. New Species of Lactarius Subg. Plinthogalus and new records of Lactifluus

subg. Gerardii (Russulaceae) from Sikkim, India. Taiwania 57(1): 37-48.

Das K, Van de Putte K, Buyck B. 2010. New or interesting Russula from Sikkim Himalaya (India).

Cryptogamie Mycologie 31(4): 373-387.

Das K, Atri NS, Buyck B. 2013. Three new species of Russula (Russulales) from India. Mycosphere

4(4): 707-717. http://dx.doi.org/10.5943/mycosphere/4/4/9

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for Basidiomycetes -

application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

130 ... Das, Verbeken, & Nuytinck

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.

Heilmann-Clausen J, Verbeken A, Vesterholt J. 1998. The genus Lactarius. Fungi of Northern

Europe, vol. 2. The Danish Mycological Society.

Henderson DM, Orton PD, Watling R. 1969. British fungus flora: agarics and boleti: colour

identification chart. Royal Botanic Garden Edinburgh, UK.

Hesler LR, Smith AH. 1979. North American species of Lactarius. USA, Ann Arbor, University of

Michigan Press.

Holmgren PK, Holmgren NH, Barnett LC. 1990. Index Herbariorum. Part 1: Herbaria of the world,

86" ed. Bronx: New York Botanical Garden, USA.

Katoh K, Standley DM. 2013. MAFFT multiple sequence alignment software version 7:

improvements in performance and usability. Molecular Biology and Evolution 30: 772-780.

http://dx.doi.org/10.1093/molbev/mst010

Kornerup A, Wanscher JH. 1981. Methuen handbook of colour. UK, London, EyreMethuen Ltd.

Reprint.

Kranzlin, F. 2005. Fungi of Switzerland. Volume 6, Russulaceae. Verlag Mykologia Luzern,

Switzerland.

Le HT, Nuytinck J, Verbeken A, Desjardin DE, Lumyong S. 2007. Lactarius in Northern Thailand: 1.

Lactarius subg. Piperites. Fungal Diversity 24: 173-224.

Nuytinck J, Verbeken A. 2003. Lactarius sanguifluus versus Lactarius vinosus molecular and

morphological analyses. Mycological Progress 2: 227-234.

http://dx.doi.org/10.1007/s11557-006-0060-5

Stamatakis A. 2006. RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

http://dx.doi.org/10.1093/bioinformatics/btl446

Van de Putte K, Nuytinck J, Stubbe D, Le HT, Verbeken A. 2010. Lactarius volemus sensu lato

(Russulales) from northern Thailand: morphological and phylogenetic species concepts

explored. Fungal Diversity 45: 99-130. http://dx.doi.org/10.1007/s13225-010-0070-0

Van de Putte K, Nuytinck J, Das K, Verbeken A. 2012. Exposing hidden diversity by concordant

genealogies and morphology—a study of the Lactifluus volemus (Russulales) species complex

in Sikkim Himalaya (India). Fungal Diversity 55: 171-194.

http://dx.doi.org/10.1007/s13225-012-0162-0

Verbeken A, Horak E. 2000. Lactarius (Basidiomycota) in Papua New Guinea. 2. Species in tropical-

montane rainforest. Austr. Syst. Bot. 13: 649-707. http://dx.doi.org/10.1071/SB99023

Verbeken A, Nuytinck J. 2013. Not every milkcap is a Lactarius. Scripta Botanica Belgica 51:

162-168.

White TJ, Bruns T, Lee SS, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal

RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR Protocols: a guide to

methods and applications. Academic Press, New York.

MYCOTAXON

http://dx.doi.org/10.5248/130.131

Volume 130, pp. 131-143 January-March 2015

Synonymy of two species of Bipolaris

from aquatic crops of Poaceae

ZI-LAN XIAO’, KEVIN D. Hype’, & JING-ZE ZHANG"

"Institute of Biotechnology, College of Agriculture & Biotechnology, Zhejiang University,

Hangzhou 310058 China

?Institute of Excellence in Fungal Research, School of Science, Mae Fah Luang University,

Chiang Rai 57100 Thailand

*CORRESPONDENCE TO: jzzhang@zju.edu.cn

ABSTRACT— Our morphological data indicate that conidia of Bipolaris zizaniae (a pathogen

causing brown spot on leaves of Zizania latifolia) are similar to those of B. oryzae. Our

sequences of the rDNA ITS, GPDH, and EF1-a gene regions from B. zizaniae and blast

searches revealed a 99-100% similarity with sequences of B. oryzae. Phylogenetic studies

also cluster B. zizaniae isolates with B. oryzae in a clade with 100% bootstrap support.

Pathogenicity testing also confirmed that B. zizaniae does infect both Zizania latifolia and

rice, causing brown spots.

Key worps—graminicolous fungi, molecular phylogeny, taxonomy

Introduction

Zizania latifolia (Griseb). Turcz. ex Stapf is a perennial grass parasitized by

the smut fungus Ustilago esculenta Henn., which causes swelling of its upper

culms. The swollen culms are used as an aquatic vegetable, commonly called

jiaobai (white bamboo) in China (Zhang et al. 2012). Due to its unique flavor

and delicacy, the demand for jiaobai has steadily increased in tandem with

the rapid economic development of China. Zhejiang Province has become

the largest Z. latifolia cultivation area in China (Xu et al. 2009). Brown spot

is one of the most severe leaf spot diseases of this crop plant. With large-scale

cultivation and changed growing conditions, brown spot incidence has become

more severe in the fields, and the disease has become a severe problem in

crop production. The pathogen causes a large number and areas of coalescing

spots (especially during seedling production in greenhouses), and these spots

expand or coalesce to form large lesions, eventually resulting withering and

death of entire leaves (Fic. 1A-D).

132 ... Xiao, Hyde, & Zhang

The pathogen causing this disease was first described as Helminthosporium

zizaniae by Nisikado (1929). Although H. zizaniae resembles H. oryzae

morphologically and by infecting Oryza sativa L. [rice], Nisikado (1929)

differentiated the two species based on the shape of the conidial base, conidial

dimensions, and conidial hilum. Shoemaker (1959) transferred H. zizaniae and

H. oryzae to Bipolaris, a placement later accepted by Shoemaker (1966) and

Sivanesan (1987). Sivanesan (1987) used conidial dimensions as an important

character in his dichotomous key to Bipolaris; he distinguished B. zizaniae

as having >150 um long conidia compared with the <150 um long conidia

in B. oryzae. However, conidial dimensions are usually variable in Bipolaris

(Sivanesan et al. 1987; Manamgoda et al. 2012), and the conidial length in

H. oryzae is not stable and can be more than 150 um in some strains (Chang

1978). Other studies showed that B. oryzae is interfertile with B. zizaniae and

that their isolates from rice and Zizania are reciprocally infective (Chang 1974;

Tsuda & Ueyama 1975, 1976). Bipolaris zizaniae and B. oryzae are now regarded

as conspecific (Manamgoda et al. 2014).

Accurate identification and precise naming of species are crucial, for species

names provide an ideal framework for storage and information retrieval for

effective disease management (Inderbitzin et al. 2011, Manamgoda et al. 2012).

Bipolaris zizaniae is an important pathogen on Z. latifolia, but due to limited

distribution, there is little information on its effects; even the morphological

descriptions of the pathogen are inadequate (Nisikado 1929, Sivanesan 1987). In

contrast, B. oryzae [sexual state previously known as Cochliobolus miyabeanus]

causes brown spots on leaves and glumes of rice, leading to serious losses in

yield (Scheffer 1997). Thus this pathogen and rice disease have been studied

worldwide (Kulkarni et al. 1980, Kubo et al. 1989, Amadioha 2002, Manandhar

et al. 1998, Xio et al. 1991). Nonetheless, the relationship between B. zizaniae

and B. oryzae is unresolved. With the use of molecular taxonomy, DNA

sequence analyses allow testing the morphological species circumscriptions

and providing new insights into species relationships, especially for complexes

of closely related, morphologically similar species (Manamgoda et al. 2012,

2014; Udayanga et al. 2012; Hyde et al. 2014). Studies of Bipolaris species have

already revealed relationships between morphological species circumscriptions

and molecular data, but there has been no study of B. zizaniae (Manamgoda et

al. 2011, 2012, 2014; Hyde et al. 2014).

We used sequences from the ITS (internal transcribed spacer), GPDH

(glyceraldehyde-3-phosphate dehydrogenase), and EFl-a (translation

elongation factor 1-a) gene regions to investigate the phylogenetic relationships

of six Bipolaris cultures isolated from Zizania latifolia in southeast China.

The objectives were to review morphological similarities in B. zizaniae and

B. oryzae and to resolve their relationship.

Bipolaris zizaniae confirmed as a synonym of B. oryzae ... 133

FiGurE 1 Symptoms of Bipolaris brown spot on Zizania latifolia and Oryza sativa. A-D. On

nature host of Z. latifolia. A. Early spots. B. Severe spots. C. Removed leaves with severe brown

spots from seedlings in the greenhouse. D. Leaf blight. E-E Symptom variability on backs of

leaves of Z. latifolia after artificial inoculation. E. Five days after inoculation. F Eighteen days

after inoculation. G-I. Symptom variability on leaves of O. sativa after artificial inoculation.

G. Brown spots eight days after inoculation. H. Brown spots on rear of leaf nine days after

inoculation. I. Brown spots twelve days after inoculation.

134 ... Xiao, Hyde, & Zhang

66S997X[

STOTOON[

€ZOTOON

PEOLOON[

SIOTOON[

86S997X[

96S997X[

PIOTOON[

€LOTOON

TTOTOONI

OLOTOONL

S00TOON[

LOOTOONL

900T09NL

COOTOON

TCOTOON[

78688910

18688911

08688914

62688914

82688910

LL68891M

985997X[

T8S99TX[

G8S99TX[

610TO9N{

LIOTOON[

18S997X[

Y-THH

SPPOLTX[

086009NI

6L6009N{

8Z6009N{

9Z6009N[

€PPOLTX[

IPP9OLTX[

0Z6009N[

696009N{

Z96009N{

996009NI

P96009NI

€96009N{

Z96009N{

9EOIOONL

LL6009N{

92688910

CL688910

FL68891M

€L688910

7L688910

1Z688914

T€v9L7Xx[

87POLTX[

OLP9L7X{

SZ6009N{

€Z6009N{

PL6009NI

LCOPOLTX[

Hdd)

‘ON ANVGNAD

"PLOT ‘TLOT ‘TLOTZ ‘Je 19 epostueueyy Wor ore soduanbas Jay}0 {UO} pjog ut poyuasaid are sasuanbas MaN ‘somnyno adéq-xa = ,

€€P9STX[

S8E76INI

L8E76IN[

O8E76INI

ICOTOON

IEP9STX[

67P9STXI

O8€Z6INI

6LET6INI

O€OTOONI

6ZOTOONI

OLET6INI

GLET6INI

€LET6INI

9ZOLOONI

PEOTOON(

02688910

69688910

89688910

9688910

99688910

9688910

LIP9STX[

€1P9STX[

9TP9STX[

€LOTLOON{

SLET6INI

ZTEOLOONI

TIPOSTXI

SLI

erpu]

eiyersny

ureds

elyersny

pueyreqy,

vsa

eiyersny

WSN Tremey

elyersny

ueystyeg

uede/

WIeUaIA,

eiyersny

eury)

puepeyy,

puepey],

puepeyy],

viyperjsny

puryesz MaN

NOILLVOOT

sADUL DIZ

syusaf srjoqosods

DADS DZKIC

snj10juos uosodoiajaH

puvdos s1ojy)

DADS DZKIC

DUALIV] X10)

DADS DZKIC

uojAjovp uopoudy

vy oft] *Z

vy Of] *Z

vyOofiD] *Z

vy Oso] *Z

vy Of] *Z

pyofiqy] vIUvzZIZ

DAIDS ‘C

DADS DZKIC

saplodys vuavjosnyy

uojAjavp uopoudy

ISO]

«x £9 9FT SAO

SLecl ddd

CS'PLE SAO,

Solel dud

c88sl did

9020-01 DONTAW.

«x 96: 0€Z SAD

09°9ST SdD

« 16 V8¢ SAO

L601 ddd

x LS ¢LT SAO

eogtec dud

c9'€6T SAO

x 60 C6I SAO

LS°CL1 SAO

x O6ZCI ddd

9TS0-€T DON TAN

STS0-€T OONTAW

VISO-€T OONTAWN

€1S0-€T OONTAW

cIs0-€T OONTAW

TIS0-€1 DON TAW

€€Z0-01 DON TAN

690-01 DONTAW.

« STZ0-0T OONTAW

85°09T SAD

el9STt dra

x 16:08¢ S€D

8c19 dWOl

ALV TOS]

vyvjnr4aqny *D

s1uosod14q *D)

viafinids ‘5

MaUaavs “7D

DIOINADAO 7)

bywuny *D

pyd1ouowoy “>

siuosodo.sajay *D

sisuanvmvy “D

DIONUIMIDAS “D

usa ‘OD

$19109 °D

sIsuUayDAJSND *D

sisuauvisasad ‘g¢

avzA1o “J

vsojnpou “g

avUavIOLIIUL “gg

puvaffoo ‘g

SHIOTdS

‘sisAjeue sduanbas WNC 9Ud8-1][NUT Ul pasn saye[Os! VLYINAIND pure s1ivjodig *| TTaVI,

Bipolaris zizaniae confirmed as a synonym of B. oryzae ... 135

Materials & methods

Collection and isolation of fungi

Diseased leaves of Zizania latifolia with brown spots growing in fields or greenhouses

were collected and returned to the laboratory. Tissue pieces (each approximately 5 x 5

mm) were cut from the margins of brown spots, between healthy and infected parts of

leaves, which were previously surface disinfected with 0.5% NaOCl. The pieces were

plated on to PDA medium and incubated at 25°C for 7 days. Mycelia from the infected

samples were transferred to fresh PDA medium. Single-spore isolation was performed

as described by Chomnuntiet al. (2014). Agar discs (with mycelium on the surface, each

approximately 5 x 5 mm) from these isolates were stored in 1.5 mL cryotubes with 20%

glycerol at -70°C.

Herbarium materials are deposited at MFLU herbarium of Mae Fah Luang University,

Chiang Rai, Thailand (MFLU). The cultures are maintained at Mae Fah Luang University

Culture Collection (MFLUCC) and Institute of Biotechnology, Zhejiang University,

Zhejiang Province, China (ZJU).

DNA extraction

Genomic DNA was extracted using a modified protocol of Zhang and Li (2009).

Fungal isolates were cultured on PDA at 25°C for 7 days. Fresh mycelia were scraped

from the surfaces of PDA plate and grounded to a fine powder in liquid nitrogen. 500 mg

power was transferred to a 1.5 mL microcentrifuge tube and 700 uL of preheated (60°C)

2x CTAB extraction buffer (2% hexadecyltri-methlammonium bromide(w/v), 100 mM

Tris-HCl, 1.4 M NaCl, 30 mM EDTA, pH 8.0) was added. The solution was incubated

in a water bath at 65°C for 60 min with a gentle swirling. Following a phenol/chloroform

extraction, the genomic DNA was precipitated by isopropanol in the presence of sodium

acetate. The precipitate was rinsed with 70% ethanol centrifuged at 12,000 rpm and

genomic DNA was resuspended in 50 uL TE buffer and stored in —20°C.

PCR amplification

The DNA fragments were amplified in an automated thermal cycler (Eppendorf AG,

Germany). Amplification was performed in a 50 uL reaction volume which contained

5 wL 10x PCR buffer, 1 uL of each primer (10 uM), 2 uL template DNA, and 0.5 uL Taq

DNA polymerase. Primers ITS6 and ITS4 (Zhang & Li. 2009) were used to amplify

the ITS and 5.8 S regions. The thermal cycling program was performed with 35 cycles

after an initial denaturation at 95°C for 4 min. Each cycle included a denaturation step

at 95°C for lmin, an annealing step at 55°C for lmin, and an extension step at 72°C

for 1.5 min. The primers gpd1/ gpd2 (Berbee et al. 1999) were used for amplifying the

GPDH gene with the changes of annealing temperature 52°C. Similarly, the EF-la gene

was amplified using primers EF 983/2218R (Schoch et al. 2009) with the changes of

annealing temperature 54°C.

Sequence alignment and phylogenetic analysis

The purified PCR products were submitted to Sunny Biotechnology Company

Limited (Shanghai, China) for sequencing in both directions. Sequence files were

assembled and edited using BioEdit software (Hall 1999). A blast search was performed

136 ... Xiao, Hyde, & Zhang

for searching the highly similar sequences in the GenBank database and related

sequences were downloaded (TABLE 1). The sequences were aligned with Clustal X 25

(Thompson et al. 1997). Sequences were analyzed phylogenetically in PAUP v4.0b10

(Swofford 2002) and trees were visualized with TreeView (Page 1996). Phylogenetic

trees were inferred from the ITS, GDPH and EF1-a sequence data set using parsimony

analysis with all characters weighted equally and all gaps treated as missing data.

Morphological studies

Conidia and conidiophores taken directly from nature hosts were examined in 3%

KOH, using a Zeiss Axiophot 2 microscopy with Axiocam CCD camera and AxioVision

digital imaging software (AxioVision Software Release 3.1.v.3-2002; Carl Zeiss Vision

Imaging Systems). Cultural characteristics and morphology were determined on PDA,

and water agar and wheat straw (WSA) [2 % water agar (10 g/L) with autoclaved wheat

straw placed onto the medium] and incubated for 7-10 days at 25°C under 12/12 h

alternation of near-UV light. 20 uL conidial suspension (10* conidia/mL) were dropped

on the WA (water agar) medium at 25°C for 24 h and characteristics of conidia

germination were observed.

Scanning electron microscopy

The 0.5 x 0.5 mm’ pieces from diseased spots with conidia and conidiophores were

placed in glutaraldehyde (2.5% vol/vol) for 48 h at 4°C. Samples were washed three times

in 0.1 M sodium phosphate buffer (pH 7.0) and post-fixed in osmium tetroxide (1% wt/

vol) for 1.5 h at 20°C. Samples were rinsed thoroughly with 0.2 M phosphate buffer (pH

6.8) and dehydrated in a graded ethanol series (30, 50, 70, 80, 90, 95, and 100%). Pieces

were embedded in Spurr’s epoxy resin. The sites containing conidia were identified

by light microscopy and ultrathin sections were cut with a Reichert-Jung Ultracut E

Ultramicrotome with a diamond knife. Ultrathin sections were collected on Formvar-

coated slot copper grids; after drying, the grids were stained with uranyl acetate and

lead citrate and examined using a transmission electron microscope (Hitachi S-3000N,

Tokyo, Japan).

Pathogenicity Tests

Healthy field cuttings of Z. latifolia with 3-5 nodes were cultivated in a 25-cm diam.

barrel with cultivated soil and placed in greenhouse at 26-28°C for one month. Oryza

sativa seedlings grown in seedbeds were transplanted into a 20-cm diam. barrel and

placed in greenhouse under the same conditions for one month.

An aqueous inoculation suspension (1 x 10° conidia/mL) with 1% vol/vol Tween-20

was prepared from 8-10-day old cultures. Pathogenicity tests were conducted on healthy

leaves of Zizania latifolia and Oryza sativa. A 50 uL conidial suspension was dropped on

two sides of each leaf and then inoculated plants were covered by large plastic bags and

incubated at 27-28°C for 48 h after which the plastic bags were removed. Control leaves

were inoculated with 50 uL sterile water. Symptoms and sizes of diseased spots were

observed and recorded each day. The fungus was re-isolated by cutting small portions

from the margin of diseased spots; these were surface sterilized and placed on PDA

plates. The experiments were repeated twice to confirm the results.

Bipolaris zizaniae confirmed as a synonym of B. oryzae ... 137

Results

Phylogenetic analysis

Isolates of Bipolaris were obtained from Z. latifolia and purified, and six

single-spore isolates were selected for study. Six strains of Bipolaris from

Z. latifolia had identical ITS and GPDH gene sequences, but in the EF-la

gene sequence there was one base difference between the two strains from

Hangzhou (MFLUCC13-0511, MFLUCC13-0512) and the other four strains

(MFLUCC13-0513, MEFLUCC13-0514, MFLUCC13-0515, MFLUCC13-0516). A

GenBank search showed that there were no sequence data submitted for B.

zizaniae, but a blast search of our ITS, GPDH, and EFl-a sequences from

Bipolaris nodulosus CBS 160.58

65 Bipolaris peregianensis BRIP 12790

400 99 | Bipolaris microlaenae CBS 280.91

Bipolaris microlaenae BRIP 15613

Bipolaris cynodontis |CMP 6128

Bipolaris oryzae MFLUCC 10-0715

Bipolaris oryzae MFLUCC 10-0694 | ftom rice

100| Bipolaris oryzae MFLUCC 10-0733

100 Bipolaris oryzae MFLUCC 13-0511

Bipolaris oryzae MFLUCC 13-0512

Bipolaris oryzae MFLUCC 13-0513 |om zizania

61 Bipolaris oryzae MFLUCC 13-0516

/7' | Bipolaris oryzae MFLUCC 13-0515

67 | Bipolaris oryzae MFLUCC 13-0514

Curvularia graminicola BRIP 23186a

86 Curvularia homomorpha CBS 156.60

Curvularia coicis CBS 192.29

Curvularia tripogonis BRIP 12375

92 Curvularia ovariicola BRIP 15882

ius Curvularia ravenelii BRIP 13165

Curvularia heteropogonis CBS 284.91

400 pCurvularia lunata MFLUCC 10-0706

Curvularia lunata CBS 730.96

60 Curvularia australiensis CBS 172.57

“ Curvularia ellisii CBS 193.62

Curvularia Spicifer CBS 274.52

400 r- Curvularia hawaiiensis BRIP 10971

Curvularia hawaliensis CBS 173.57

Curvularia tuberculata CBS 146.63

_10

FiGuRE 2. Phylogram generated from parsimony analysis based on ITS, GPDH and EF 1-a

sequence data of Bipolaris and Curvularia species. Data were analysed with random sequence

addition, unweighted parsimony and by treating gaps as missing data. Bootstrap values =>50% are

indicated. Species names are followed by the number of the deposited voucher. The tree is rooted

with Curvularia tuberculata.

138 ... Xiao, Hyde, & Zhang

B. zizaniae showed a 99-100% similarity with those of B. oryzae lodged in

GenBank. Of the 63 reliable or voucher B. oryzae ITS sequences in GenBank,

11 were identical with B. zizaniae and 41 differed by 1-2 bases. Similarly, of

the 16 reliable or voucher B. oryzae GPDH sequences in GenBank, one was

identical with B. zizaniae and 15 differed by 1-2 bases. Finally, of five B. oryzae

EF1-a sequences (Manamgoda et al. 2012), four were identical with B. zizaniae

and one differed by 1-2 bases.

A parsimony tree was constructed from the ITS, GPDH, and EF1-a data

from 29 fungal isolates (TABLE 1). The four Bipolaris isolates from Z. latifolia

with identical sequences grouped together with another two Bipolaris isolates

from Z. latifolia (with one base difference) with 61% bootstrap value support

(Fic. 2). This group clustered with three B. oryzae isolates with a 100% bootstrap

support. Fic. 2 clearly shows that the Bipolaris isolates from Z. latifolia are

inseparable from B. oryzae. Sequence variation between Bipolaris isolates

from Z. latifolia and B. oryzae isolates was less than interspecific variation.

Six Bipolaris isolates from Z. latifolia and three B. oryzae isolates clustered

with B. coffeana Sivan. to form a clade with a 70% bootstrap value support

(Fic. 2). However, the B. coffeana subclade (based on a single culture) and the

B. oryzae/B. zizaniae subclade were separated by >40 base differences, indicating

that they are not conspecific.

The description below is based on our collections of Bipolaris isolates from

Z. latifolia. Except for size variation, these characters are almost identical to

those from the type specimen from the same host from Japan (Nisikado 1929).

The pathogen causes brown spots on leaves of Zizania latifolia and infects

almost all aerial parts of the plant.

Bipolaris oryzae (Breda de Haan) Shoemaker, Can. J. Bot. 37: 883 (1959) Fig23

= Helminthosporium oryzae Breda de Haan, Bulletin Inst. Bot. Buitenzorg 6: 11 (1900)

= Ophiobolus miyabeanus S. Ito & Kurib., Ann. Phytopath. Soc. Japan 2(1):

7 (1927)

= Cochliobolus miyabeanus (S. Ito & Kurib.) Drechsler ex

Dastur, Indian. J. Agr.Res. 12: 733 (1942)

= Helminthosporium zizaniae Y. Nisik., Ber. Ohara Inst.

Landw. Forsch. Kurashiki 4: 122 (1929)

= Bipolaris zizaniae (Y. Nisik.) Shoemaker, Can. J. Bot. 37: 885 (1959)

Leaf spots dark brown with a paler centre 0.3-0.5 x 0.3-0.6 cm. On a natural

host, conidiophores single or occasionally fasciculate in small groups, rarely

branched, geniculate or straight in sterile part, then becoming slightly to

distinctly geniculate, cicatrized with scars often inflated at base, medium

olivaceous-brown below, paler at the apex, multiseptate, 106-395 um long,

swollen to 7-9 um diam at base, then narrowing to 6-7 um diam (middle)

Bipolaris zizaniae confirmed as a synonym of B. oryzae ... 139

FiGurE 3. Bipolaris oryzae (isolates from Z. latifolia). A-D. Light microscope micrographs.

A. Conidia. B. Conidia on WSA. C. Conidiophore. D. Germinated conidium with one germ tube at

each end. Basal germ tube direction of growth (left below) is semiaxial, close to hilum. E-G. SEM

micrographs. E. Conidia. F. Hilum. G. Germinated conidium with two polar germ tubes. Scale bars:

A, B, D(lower) = 10 um; C, D(upper) = 20 um; E= 15 um; F=5 um; G= 25 um.

and 3-5 um (apex). Conidia (65-)78-110(-131) x (15-)17-28(-34) um,

6-10 distoseptate, olivaceous brown or yellowish brown, obclavate, navicular,

straight or slightly curved, tapering towards the apex, hilum dark, conspicuous,

sometimes slightly protruding. Conidia germinate by the production of two

polar germ tubes. The sexual morph is not found in nature.

On WSA, conidiophores 212-646 x 5.5-10.5 um, olivaceous-brown, paler

at the apex. Conidia brown or dark brown, obclavate, fusoid, often curved,

(73-)80-125(-130) x (1-)18-28(-31) um, 6-11-distoseptate, hilum dark,

conspicuously protruding or truncate.

MATERIALS EXAMINED: CHINA, ZHEJIANG PROVINCE, Hangzhou Suburbs,

September 2012, Zilan Xiao (MFLU13-0092; cultures MFLUCC13-0511 = ZJU0101,

MFLUCC13-0512 = ZJU0102); ANHUI PROVINCE, Yuexi County, September 2012,

Zilan Xiao (MFLU13-0093; cultures MFLUCC13-0513 = ZJU0201, MFLUCC13-0514

= ZJU0202); Fus1AN PROVINCE, Putian city, October 2012, Zilan Xiao (MFLU13-0094;

cultures MFLUCC13-0515 = ZJU0301, MFLUCC13-0516 = ZJU0302).

140 ... Xiao, Hyde, & Zhang

TABLE 2. Synopsis of conidial size and number of distosepta in Bipolaris oryzae from

Zizania latifolia from different origins.

ISOLATE LENGTH (um) WIDTH (um) DISTOSEPTA

ON HosTS MEFLU 13-0092 99 +11.5 25 + 3.0 (5-)7-9

MEFLU 13-0093 87 + 8.7 20 + 3.6 (5-)7-10

MEFLU 13-0094 92 + 10.6 2343.5 (5-)6-10

On WSA MFLUCC 13-0511 96 +11 24+3.8 6-9(-11)

MFLUCC 13-0513 86 + 7.8 19+5.4 6-10

MFLUCC 13-0515 90 + 5.6 2146.7 6-10(-11)

CoMMENTS—Classification in the genus Bipolaris was previously based

on morphology, with conidial characters being considered as important in

species delimitation. In this study conidia of Bipolaris isolates from Z. latifolia

are shorter (78-110 um) and wider (17-28 um) than those reported in the

protologue (25-166 x 12-13 um; Nisikado 1929). Although there is some

variation in conidial size and distosepta in Bipolaris isolates from Z. latifolia

from different locations in China (TaBLE 2), the morphological differences are

not significant. We found it impossible to distinguish B. oryzae from B. zizaniae

based on morphological characters.

Morphology and molecular phylogeny confirm that Bipolaris zizaniae and

B. oryzae are conspecific. Bipolaris oryzae is the correct name for this expanded

taxon because the epithet oryzae (published in 1900) has priority over zizaniae

(published in 1929). We follow Manamgoda et al. (2012, 2014) in preferring

Bipolaris to the earlier generic name Cochliobolus (proposed for the sexual

state).

Pathogenicity testing

The isolates of Bipolaris isolated from Z. latifolia infected the leaves of

Z. latifolia. At 48 h after inoculation, Bipolaris isolates from Z. latifolia induced

small flecks or expanded spots on the lower surface of Z. latifolia leaves.

Symptoms were similar to those observed in the wild (Fic. 1£,F), and no

disease was found on upper surface of inoculated or control leaves. Five days

after inoculation, diseased spots increased in size (Fic. 1E). Symptoms of leaf

blight occurred 18 days after inoculation (Fic. 1F). During the same period,

disease symptoms did not develop on the upper surface or on control leaves.

Similarly, isolates of Bipolaris isolated from Z. latifolia also infected rice

leaves. Small flecks occurred on the upper surface of inoculated leaves after

five days and similar flecks occurred on the lower surface of leaves after six

days. After 8-9 days, diseased spots increased more rapidly in size on the upper

surface of leaves than on the lower surface of leaves (Fic. 1G, H). The diseased

spots were approximately 0.5 cm in diameter 12 days after inoculation (Fie. 11).

Bipolaris zizaniae confirmed as a synonym of B. oryzae... 141

Discussion

Zizania latifolia, which originated in the northern region of China, was

once used as an important grain in ancient times. Cultivated Z. latifolia was

disseminated to Taihu Lake from north to south, and the Taihu Lake basin

is the region associated with the origin of jiaobai (Xu et al. 2008). Jiaobai

cultivation was introduced to eastern and southeastern Asia, including the

Russian Far East, Japan, and Korea (Thrower & Chan 1980, Guo et al. 2007).

It is possible that brown spot was also introduced to the jiaobai-growing areas

with the planting material.

Zizania latifolia commonly grows in shallow water and can be found at the

edges of lakes, ponds, wetlands, and flooded fields (Thrower & Chan 980, Guo

et al. 2007). In some cases, Z. latifolia is able to grow in deeper water (Yamasaki

& Tangel981), because it has a well-developed ventilation system (Yamasaki

1984, 1987). Thus, Z. latifolia growing conditions are similar to that of rice.

In the jiaobai-growing areas, Z. latifolia is usually grown in paddy fields or

in rotation with rice. The fact that pathogenic brown spot of Z. latifolia also

infects rice may be a result of long-term evolution. In morphological taxonomy,

phenotypic plasticity and homoplasy may influence fungal species delimitation

(Pino-Bodas et al. 2011). Bipolaris zizaniae and B. oryzae have previously been

delineated based on morphology, although species boundaries are not clear

(Chang 1974; Tsuda & Ueyama 1975, 1976). Bipolaris zizaniae can be crossed

with B. oryzae to form the sexual state, and both species can infect Zizania

(Chang 1974, Tsuda et al. 1975). The data presented here and in previous studies

suggest that B. oryzae and B. zizaniae are synonymous. Our gene sequence data

analysis shows unequivocally that B. oryzae and B. zizaniae are conspecific.

Acknowledgments

The authors are grateful to Dimuthu $. Manamgoda (School of Science, Mae Fah

Luang University, Thailand) and Lei Cai (State Key Laboratory of Mycology, Institute

of Microbiology, Chinese Academy of Sciences, People’s Republic of China) for their

helpful comments on this manuscript. The research was funded by the National Natural

Science Foundation of China (No. 31070123) and the Special Fund for Agro-scientific

Research in the Public Interest of China (No. 201003004). KD Hyde thanks MFLU grant

No. 56101020032 for funds to study Dothideomycetes.

Literature cited

Amadioha AC. 2002. Fungi toxic effects of extracts of Azadirachta indica against Cochliobolus

miyabeanus causing brown spot disease of rice. Archives of Phytopathology and Plant

Protection 35: 37-42. http://dx.doi.org/10.1080/032354002 1000009597

Berbee M, Pirseyedi M, Hubbard S. 1999. Cochliobolus phylogenetics and the origin of known,

highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase

gene sequences. Mycologia 91: 964-977. http://dx.doi.org/10.2307/3761627

142 ... Xiao, Hyde, & Zhang

Chang HS. 1974. Intercross fertility between Helminthosporium oryzae, H. zizaniae and an

unidentified Helminthosporium sp. on Zizania aquatica. Botanical Bulletin of Academia Sinica

15: 103-111.

Chang HS. 1978. A elongated conidium strain and mating type distribution of Cochliobolus

miyabeanus. Botanical Bulletin of Academia Sinica 19: 139-144.

Chomnunti P, Hongsanan S, Aguirre-Hudson B, Tian Q, PerSoh D, Dhami MK, Alis AS,

Xu J, Liu X, Stadler M, Hyde KD. 2014. The sooty moulds. Fungal Divers 66:1-36

http://dx.doi.org/10.1007/s13225-014-0278-5

Guo HB, Li SM, Peng J, Ke WD. 2007. Zizania latifolia Turcz. cultivated in China. Genetic Resources

and Crop Evolution 54: 1211-1217. http://dx.doi.org/10.1007/s10722-006-9102-8

Hall TA. 1999. Bioedit: a user-friendly biological sequence alignment editor and analysis program

for Windows 95/98/NT. Nucleic acids symposium series 41: 95-98.

Hyde KD, Nilsson RH, Alias SA, Ariyawansa HA, Blair JE, Cai L, de Cock AWAM, Dissanayake

AJ, Glockling SL, Goonasekara ID, Gorczak M, Hahn M, Jayawardena RS, van Kan JAL,

Laurence MH, Lévesque CA, Li X, Liu JK, Maharachchikumbura SSN, Manamgoda DS,

Martin FN, McKenzie EHC, McTaggart AR, Mortimer PE, Nair PVR, Pawlowska J, Rintoul

TL, Shivas RG, Spies CFJ, Summerell BA, Taylor PWJ, Terhem RB, Udayanga D, Vaghefi N,

Walther G, Wilk M, Wrzosek M, Xu JC, Yan JY, Zhou N. 2014. One stop shop: backbones

trees for important phytopathogenic genera: I (2014). Fungal Divers. 67(1): 21-125.

http://dx.doi.org/10.1007/s13225-014-0298-1

Inderbitzin P, Bostock RM, Davis RM, Usami T, Platt HW, Subbarao KV. 2011. Phylogenetics and

taxonomy of the fungal vascular wilt pathogen Verticillium, with the descriptions of five new

species. PLoS ONE 6: e28341. http://dx.doi.org/10.1371/journal.pone.0028341

Kubo Y, Tsuda M, Furusawa I, Shishiyama J. 1989. Genetic analysis of genes involved in

melanin biosynthesis of Cochliobolus miyabeanus. Experimental mycology 13: 77-84.

http://dx.doi.org/10.1016/0147-5975(89)90010-8

Kulkarni S$, Ramakrishnan K, Hegde RK. 1980. Ecology, epidemiology, and supervised control of

rice brown leaf. International Rice Research Newsletter 5: 13-14.

Manamgoda DS, Cai L, Bahkali AH, Chukeatirote E, Hyde KD. 2011. Cochliobolus: an overview

and current status of species. Fungal Diversity 51: 3-42.

http://dx.doi.org/10.1007/s13225-011-0139-4

Manamgoda DS, Cai L, McKenzie EHC, Crous PW, Madrid H, Chukeatirote E, Shivas RG, Tan YP,

Hyde KD. 2012.A phylogenetic and taxonomic re-evaluation of the Bipolaris - Cochliobolus -

Curvularia complex. Fungal Diversity 56: 131-144.

http://dx.doi.org/10.1007/s13225-012-0189-2

Manamgoda DS, Rossman AY, Castlebury LA, Crous PW, Madrid H, Chukeatirote E, Hyde KD.

2014. The genus Bipolaris. Studies in Mycology 79: 221-288.

http://dx.doi.org/10.1016/j.simyco.2014.10.002

Manandhar HK, Jorgensen HJL, Mathur SB, Smedegaard-Petersen V. 1998. Suppression of rice

blast by preinoculation with a virulent Pyricularia oryzae and the non-rice pathogen Bipolaris

sorokiniana. Phytopathology 88: 735-739. http://dx.doi.org/10.1094/PHY TO.1998.88.7.735

Nisikado Y. 1929. Studies on the Helminthosporium diseases of Gramineae in Japan. Berichte des

Ohara Instituts fir Landwirthschaftliche Forschungen in Kurashiki 4: 111-126.

Page RDM. 1996. Treeview. An application to display phylogenetic trees on personal computer.

Computer Applications in the Biological Sciences 12: 357-358.

Pino-Bodas R, Rosa Burgaz A, Martin MP, Lumbsch HT. 2011. Phenotypical plasticity and homoplasy

complicate species delimitation in the Cladonia gracilis group (Cladoniaceae, Ascomycota).

Organisms Diversity & Evolution 11: 343-355. http://dx.doi.org/10.1007/s13127-011-0062-2

Bipolaris zizaniae confirmed as a synonym of B. oryzae... 143

Scheffer RP .1997. The nature of disease in plants. Cambridge University Press.

Schoch C, Crous PW, Groenewald J, et al. 2009. A class-wide phylogenetic assessment of

Dothideomycetes. Studies in Mycology 64: 1-15. http://dx.doi.org/10.3114/sim.2009.64.01

Shoemaker RA. 1959. Nomenclature of Drechslera and Bipolaris, grass parasites segregated from

Helminthosporium. Canadian Journal of Botany 37: 879-887. http://dx.doi.org/10.1139/b59-073

Shoemaker RA. 1966. A pleomorphic parasite of cereal seeds, Pyrenophora semeniperda. Canadian

Journal of Botany 44: 1451-1456. http://dx.doi.org/10.1139/b66-160

Sivanesan A. 1987. Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohilum and

their teleomorphs. CAB International.

Swofford D. 2002. PAUP 4.0 b10: Phylogenetic analysis using parsimony. Sinauer Associates,

Sunderland.

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997 The CLUSTAL_X windows

interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.

Nucleic Acids Research 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Thrower LB, Chan YS. 1980. Gau sun: a cultivated host-parasite combination from China.

Economic Botany 34: 20-26. http://dx.doi.org/10.1007/BF02859552

Tsuda M, Ueyama A. 1975. Identity of Helminthosporium-leaf spot fungi attacking Oryzoideae

plants growing in Japan Islands (preliminary note). Transaction of the Mycological Society of

Japan 16: 93-94.

Tsuda M, Ueyama A. 1976. Distribution of two mating types of Cochliobolus miyabeanus in field

and a laboratory attempt to produce hybrids between isolates of Helminthosporium oryzae from

the U.S.A. and Japan. Annals of the Phytopathological Society of Japan 42: 7-11.

http://dx.doi.org/10.3186/jjphytopath.42.7

Udayanga D, Liu X, Crous PW, McKenzie EH, Chukeatirote E, Hyde KD. 2012. A multi-

locus phylogenetic evaluation of Diaporthe (Phomopsis). Fungal Diversity 56: 157-171.

http://dx.doi.org/10.1007/s13225-012-0190-9

Xio JZ, Tsuda M, Doke N, Nishimura S. 1991. Phytotoxins produced by germination spores of

Bipolaris oryzae. Phytopathology 81: 58-64. http://dx.doi.org/10.1094/Phyto-81-58

Xu C, Hu MH, Guo DP. 2009. Current status and prospects of aquatic vegetable industry in

Zhejiang province. ChangJiang Veget 16: 106-109.

Xu X, Ke W, Yu X, Wen J, Ge S. 2008. A preliminary study on population genetic structure

and phylogeography of the wild and cultivated Zizania latifolia (Poaceae) based on ADH1A

sequences. Theoretical and Applied Genetics 116: 835-843.

http://dx.doi.org/10.1007/s00122-008-0717-3

Yamasaki S. 1984. Role of plant aeration in zonation of Zizania latifolia and Phragmites australis.

Aquatic Botany 18: 287-297. http://dx.doi.org/10.1016/0304-3770(84)90070-6

Yamasaki S. 1987. Oxygen demand and supply in Zizania latifolia and Phragmites australis. Aquatic

Botany 29: 205-215. http://dx.doi.org/10.1016/0304-3770(87)90016-7

Yamasaki S, Tange I. 1981. Growth responses of Zizania latifolia, Phragmites australis

and Miscanthus sacchariflorus to varying inundation. Aquatic Botany 10: 229-239.

http://dx.doi.org/10.1016/0304-3770(81)90025-5

Zhang JZ, Li MJ. 2009. A new species of Bipolaris from the halophyte Sesuvium portulacastrum

in Guangdong Province, China. Mycotaxon 109: 289-300. http://dx.doi.org/10.5248/109.289

Zhang JZ, Chu FQ, Guo DP, Hyde KD, Xie GL. 2012. Cytology and ultrastructure of interactions

between Ustilago esculenta and Zizania latifolia. Mycological Progress 11: 499-508.

http://dx.doi.org/10.1007/s11557-011-0765-y

MY COTAXON

http://dx.doi.org/10.5248/130.145

Volume 130, pp. 145-164 January-March 2015

Myxomycetes of Sonora (Mexico) 6.

Central plains of the Sonoran Desert

Marcos LIZARRAGA ', GABRIEL MORENO ?,, MARTIN ESQUEDA 3,

CYNTHIA SALAZAR-MARQUEZ 3, & MARTHA L. CORONADO 4

' Dpto. Ciencias Quimico Biologicas, Instituto de Ciencias Biomédicas, Univ. Autonoma de Ciudad Judrez,

Anillo Envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32300, México

? Dpto. Ciencias de la Vida, Edificio de Biologia, Univ. Alcala, Madrid, 28805, Espana

> Centro de Investigacion en Alimentacion y Desarrollo,

A.C. Apartado Postal 1735, Hermosillo, Sonora, 83304, México

‘Univ. Estatal de Sonora. Apartado Postal 11, Admon. 11, Hermosillo 83000, Sonora, México

* CORRESPONDENCE TO: gabriel.moreno@uah.es

ABSTRACT — Myxomycetes from 10 localities in the central plains of the Sonoran desert were

studied using the moist chamber culture technique. Thirty-three species have been identified,

of which five (Comatricha mirabilis, Cribraria confusa, Didymium eremophilum, Perichaena

luteola, and P. stipitata) represent new records for Sonora, and Comatricha mirabilis is cited

for the first time from Mexico. Light and scanning electron microphotographs illustrate the

most diagnostic characters of these species, which are rare or scarcely mentioned in the

literature.

Key worps — Amoebozoa, chorology, myxobiota, slime moulds, taxonomy

Introduction

The Sonoran desert, which lies in northwestern Mexico and the southwestern

United States, covers a 260,000-km2 area, with the largest portion occurring in

Sonora state (Mexico) and the rest in southwestern Arizona and southeastern

California (United States). This is the warmest and the most arid North

American desert due to its low elevation (<600 m) and 22-24 °C average annual

temperature. The bi-seasonal (winter and summer) rainfall averages 200-400

mm per year (Hernandez 2006).

The desert vegetation is characterized by trees and shrubs, including

Parkinsonia florida (A. Gray) S. Watson, Olneya tesota A. Gray, Prosopis

glandulosa Torr., Fouquieria macdougalii Nash, and Bursera spp. The majority

of the cacti are large, with Carnegiea gigantea (Engelm.) Britton & Rose the

146 ... Lizarraga & al.

TABLE 1. Studied Sonoran localities and types of vegetation

LOCALITY LATITUDE N LONGITUDE W ALTITUDE VEGETATION

Town of Benjamin Hill

1. Las Animas 30°12’37.95” 111°18’55.04” 770m MDS

Town of Opodepe

2. Tuape 30°02’52.89” 111°00’22.05” 670m M

Town of Guaymas

3. Cafién de Nacapule 27°59'08.54” 111°02’06.40” 81m SC

4. El Tigre 28°06'31.14” 111°01’21.11” 139m SC

5. El Apache 28°19'25.33” 111°14’27.74” 43m M

6. Maytorena 28°13'32.49” 110°48’50.28” 40m SC

Town of Carbo

7. San Luis 29°33’42.38” 111°04’52.79” 458 m MDS

Town of Hermosillo

8. La Primavera 28°48’06.37” 111°09’09.62” 167 m MDS

9. La Pintada 28°39'56.35” 110°57'15.64” 233 m SC

10. El Papalote 29°12'56.14” 111°02’39.43” 348 m M

Types of vegetation: MDS = microphyllous desert scrub; SC = sarcocaule scrub; M = mezquital

most representative (Hernandez 2006). Myxomycetes can develop in arid

environments due to decomposition of and water retained by succulent plants

such as Opuntia spp. These provide an excellent microhabitat with enough

moisture for optimal development. Although the Sonoran desert has a high

level of biological diversity, here have been only a few studies of its myxobiota.

The Sonoran desert records of Evenson (1961) and Blackwell & Gilbertson

(1980a,b; 1984) were obtained mainly in Arizona (U.S.A.) in the United States.

Although there have been various diverse studies of the myxobiota in Sonora

(Mexico), only twenty-seven reports are known for the biosphere reserve

El Pinacate and Gran Desierto de Altar (Esqueda et al. 2013, Moreno et al.

2004); most studies have been centered in pine-oak forests and lowland areas

(Pérez-Silva et al. 2001; Moreno et al. 2006; Lizarraga et al. 2007, 2008a,b).

The catalogue of myxomycetes for Sonora state (Moreno et al. 2007) has

been increased from 77 species to 108 species by Lizarraga et al. (2007, 2008a,b)

and Esqueda et al. (2013). The present contribution brings the total number to

Lis,

Materials & methods

Four surveys were conducted in 2009, one each season of the year, in 10 localities

of the central plains of the Sonoran desert (TABLE 1). Specimens were gathered or

cultured in moist chamber cultures. Permanent slides were prepared in Hoyer’s liquid

for microscopic study. Collections have been deposited in the collection of fungi of the

Myxomycetes of Sonora 6 (Mexico) ... 147

Universidad Estatal de Sonora, Hermosillo, Sonora, México (UES), with a small part in

the herbarium of the Universidad de Alcala, Alcala de Henares, Madrid, Spain (AH).

The spore ornamentation was examined under SEM using the material deposited in

AH. Collector names are abbreviated as follows: AG = A. Gutiérrez; ML = M. Lizarraga;

CS = C. Salazar; and AS = A. Sanchez. New species records for Sonora are marked with a

single asterisk, and the species recorded as new for Mexico is marked with two asterisks.

Taxonomy

Arcyria cinerea (Bull.) Pers.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 4, on Forchhammeria watsonii Rose,

leg. ML, CS, AS & AG, 19-VIII-2009, obtained from moist chamber, 30-IX-2009 (UES

8539); obtained from moist chamber, 9-X-2009 (UES 8541).

ComMENTS— Previously cited for Sonora by Lizarraga et al. (2007, 2008b).

Arcyria denudata (L.) Wettst.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on decomposing wood, leg. AG & AS,

22-V-2009, obtained from moist chamber, 11-VI-2009 (UES 8106).

CoMMENTS—Previously recorded for Sonora by Pérez-Silva et al. (2001), and

Lizarraga et al. (2008a,b).

Badhamia gracilis (T. Macbr.) T. Macbr.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Cylindropuntia sp., leg. AS & AG,

9-III-2009, obtained from moist chamber, 20-IV-2009 (UES 7858); Loc. 2, on Opuntia

sp., leg. AS & AG, 14-II-2009, obtained from moist chamber, 15-IV-2009 (UES 7854);

Loc. 3, on Stenocereus thurberi (Engelm.) Buxb., leg. AS & AG, 26-V-2009, obtained

from moist chamber, 30-VI-2009 (UES 8206); Loc. 4, on Lophocereus schottii (Engelm.)

Britton & Rose, leg. CS, AS & AG, 7-XI-2009, obtained from moist chamber, 4-XII-2009

(UES 8660); Loc. 5, on Parkinsonia sp., obtained from moist chamber, 20-IV-2009 (UES

7856); Loc. 6, on Cylindropuntia versicolor (Engelm.) EM. Knuth, leg. AG & AS, 12-

III-2009, obtained from moist chamber, 6-IV-2009 (UES 7845); Loc. 7, on Stenocereus

thurberi, obtained from moist chamber, 14-IV-2009 (UES 7962); Loc. 8, on Lophocereus

schottii, leg. AS & AG, 14-III-2009, obtained from moist chamber, 14-IV-2009 (UES

7847); Loc. 9, on Cylindropuntia arbuscula (Engelm.) EM. Knuth, leg. AG & AS 26-V-

2009, obtained from moist chamber, 11-VI-2009 (UES 8110); Loc. 10, on Parkinsonia

sp. leg. AS & AG, 12-III-2009, obtained from moist chamber, 20-IV-2009 (UES 7860).

COMMENTS— We follow the taxonomy of Moreno & Oltra (2010), who regarded

Badhamia melanospora Speg. as separate from B. gracilis. The two species differ

mainly in spore morphology, with the spores >15-17 um diam., polyhedral,

and densely warty in B. melanospora and <12-14 um diam, polyhedral, and

with scattered warts with intervening spaces in B. gracilis. Both species fruit

mainly on succulent plants (Castillo et al. 1996). Some studied collections were

mistakenly identified as B. melanospora by Moreno et al. (2006) and Esqueda

et al. (2013).

148 ... Lizarraga & al.

Badhamia spinispora (Eliasson & N. Lundg.) H.W. Keller & Schokn. FIGS 1, 5-6

Fructifications sessile, subglobose to short plasmodiocarps, white to

grey, growing grouped together. Sporotheca 0.2-1 x 0.2-0.5 mm. Peridium

double, calcareous, with irregular dehiscence. Hypothallus inconspicuous.

Pseudocolumella irregular, calcareous. Capillitium scarce and joined by

strands consisting of irregular calcareous nodules. Spores (9-)10-12 x 13-15

um, globose to subglobose, black in mass, violaceous with a paler area under

LM, ornamented with large spines. SEM shows that the spore ornamentation is

formed by irregularly scattered or grouped <0.5 um high warts.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 9, over hare manure, leg. AG & AS, 26-V-

2009; obtained from moist chamber, 30-VI-2009 (UES 8116, AH 42974).

ComMENTS— This species is characterized by its generally coprophilous habitat

(although it does occasionally grow on tree bark), cespitose habit, capillitium

that varies from physaroid to badhamioid, and spores that are dark in mass,

spiny, and with a paler area. A study of B. spinispora, Physarum apiculosporum

Hark., and Badhamia verrucospora G. Moreno & al. was published by Moreno

et al. (2011).

Cited from Sonora by Esqueda et al. (2012, 2013).

Comatricha elegans (Racib.) G. Lister

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AG & AS, 22-V-

2009, obtained from moist chamber, 3-VI-2009 (UES 8120); Loc. 7, leg. ML, CS, AS &

AG, 17-VIII-2009, obtained from moist chamber, 7-IX-2009 (UES 8563); Loc. 9, 20-

VIII-2009, obtained from moist chamber, 9-IX-2009 (UES 8621); Loc. 10, decomposing

wood, 21-V-2009, obtained from moist chamber, 11-VI-2009 (UES 8122).

CoMMENTS—Lizarraga et al. (2005) published a study (including SEM

photomicrographs) of C. elegans from Chihuahua.

Previously recorded from Sonora by Lizarraga et al. (2007, 2008a).

**Comatricha mirabilis R.K. Benj. & Poitras Figs 7-12

Fructifications stipitate, 0.7-0.9 mm tall. Sporotheca subglobose to

subcylindrical, 0.4-0.6 mm diam. Peridium fleeting and persistent at the

base, resembling a necklace. Stipe short, dark, 0.3-0.6 mm high. Hypothallus

membranous, hyaline, as common in Comatricha. Columella crossing the

sporotheca and forking into several branches at the apex. Capillitium parallel

along the columella, dark brown with free ends, dichotomous, widened and

forked at the tips that protrude at the periphery of the sporotheca. Spores dark

brown in mass, 9-12 um diam., globose to subglobose, reticulate, with walls

1-2 um high. SEM observation reveals that the spore is reticulate and the high

banded reticulum has entire and unperforated walls.

Myxomycetes of Sonora 6 (Mexico) ... 149

Ficures 1-6. Badhamia spinispora (AH 42974): 1. Plasmodiocarps. 5-6. Spores (SEM). Cribraria

confusa (AH 42976): 2-4. Sporocarps. Bars: 1 = 1 mm; 2-4 = 0.5 mm; 5-6 = 2 um.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 4, on bark of Forchhammeria watsonii,

leg. AG & AS, 27-V-2009, obtained from moist chamber, 30-VI-2009 (UES 8129, AH

42975).

CoMMENTS—Comatricha mirabilis is characterized by its capillitium that

extends parallel along the columella without forming a network with bifurcated

ends that protrude from the sporotheca and fully reticulate globose spores. It

was described growing over goat manure in USA in a moist chamber (Benjamin

& Poitras 1950). Later, Mandell & Blackwell (2008) gathered abundant material

from tree barks of Tamarix arabica, Prosopis juliflora, Phoenix dactylifera, and

Cordia myxa from a Bahrain desert in the Kingdom of Bahrain archipelago

near Saudi Arabia, which they developed in a moist chamber. Andrade-Bezerra

et al. (2008) described material developed on Brazilian cottontail (Sylvilagus

brasiliensis L.) dung in a moist chamber, Poulain et al. (2011) gathered it from

France on wood debris, and Schnittler et al. (2013) identified the species from

arid areas in China. We obtained three sporocarps on Forchhammeria watsonii

bark in a moist chamber. This is the first record for Mexico.

150 ... Lizarraga & al.

Comatricha tenerrima (M.A. Curtis) G. Lister

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AS & AG, 22-V-

2009, obtained from moist chamber, 15-VI-2009 (UES 8124); Loc. 9, on decomposing

wood, leg. CS, AS & AG, 6-XI-2009, obtained from moist chamber, 11-XII-2009 (UES

8685); Loc. 10, on decomposing wood, leg. AG & AS, 21-V-2009, obtained from moist

chamber, 6-VI-2009 (UES 8130).

CoMMENTS— Lizarraga et al. (2007) published a description with SEM

micrographs of C. tenerrima from Sonora. It has also been cited from other

natural protected areas in Sonora (Lizarraga et al. 2008a,b; Esqueda et al. 2013).

*Cribraria confusa Nann.-Bremek. & Y. Yamam. Fras. 2-4

Fructifications small (0.3-0.9 mm high), stipitate. Sporotheca globose to

subglobose, 0.1-0.2 mm diam. Stipe 0.2-0.7 mm long, dark brown, with debris.

Peridium evanescent, forming a small discoid calyculus at the base beneath

an open network of irregular flattened nodules covered by dictydine granules.

Hypothallus membranous. Spores 7-8 um diam., yellow in mass, light yellow

under LM, globose, with a warty ornamentation.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 3, on palm fibers, leg. ML, CS, AS &

AG, 19-VIII-2009, obtained from moist chamber, 5-I[X-2009 (UES 8570); obtained

from moist chamber, 7-IX-2009 (UES 8759, AH 42976 with Arcyria cinerea and

Paradiacheopsis sp.); on Guaiacum coulteri A. Gray, obtained from moist chamber, 3-X-

2009 (UES 8571).

ComMMENtTS—Cribraria confusa has been recorded only from Chihuahua on

decomposing wood of Pinus sp. by Lizarraga et al. (2003), who presented LM

and SEM micrographs.

Cribraria violacea Rex

STUDIED MATERIAL: MEXICO. Sonora: Loc. 10, on palm fibers, leg. AG, 21-V-2009,

obtained from moist chamber, 15-VI-2009 (UES 8135).

CoMMENTS— Moreno et al. (2001) published an SEM study of the spore

ornamentation on material from Baja California.

Previously cited from Sonora by Moreno et al. (2006) and Lizarraga et al.

(2008b).

Dictydiaethalium plumbeum (Schumach.) Rostaf.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on decomposing wood, leg. ML, CS, AS

& AG, 17-VIII-2009, obtained from moist chamber, 9-IX-2009 (UES 8572); Loc. 5, leg.

AG & AS, 14-III-2009, obtained from moist chambers, 1-V-2009 (UES 7880).

CoMMENTS—Previously cited from Sonora by Moreno et al. (2006) and

Lizarraga et al. (2008b).

Myxomycetes of Sonora 6 (Mexico) ... 151

FIGURES 7-17. Comatricha mirabilis (AH 42975): 7. Sporocarp. 8. Capillitium. 9. Spores.

10. Capillitium endings (detail, SEM). 11-12. Spores by SEM. Didymium eremophilum (AH

42977): 13. Sporocarps. 14. Sporocarp (detail). 15-17. Variability of spore ornamentation (SEM).

Bars: 7 = 0.1 mm; 8 = 50 um; 9 = 10 um; 13-14 = 0.5 mm; 15 = 5 um; 16-17 = 2 um.

152 ... Lizarraga & al.

Didymium dubium Rostaf.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AS & AG,

9-III-2009, obtained from moist chamber, 20-IV-2009 (UES 7888); Loc. 2, on Prosopis

glandulosa, leg. ML, CS, AS & AG, 19-VII-2009, obtained from moist chamber, 9-IX-

2009 (UES 8755); Loc. 5, on Prosopis glandulosa, leg. AS & AG, 14-III-2009, obtained

from moist chamber, 9-IV-2009 (UES 7886); Loc. 6, on Prosopis glandulosa, leg. AG

& AS 12-III-2009, obtained from moist chamber, 25-IV-2009 (UES 7889); Loc. 8, on

Cylindropuntia sp., leg. AG & AS, 14-III-2009, obtained from moist chamber, 14-IV-

2009 (UES 7882); Loc. 9, on Cylindropuntia arbuscula, leg. CS, AS & AG, 6-XI-2009,

obtained from moist chamber, 26-XI-2009 (UES 8692).

ComMMENTS—Moreno et al. (2001) published a SEM study of the spore

ornamentation. Previously cited from Sonora by Moreno et al. (2006).

*Didymium eremophilum M. Blackw. & Gilb. Fics 13-17

Fructifications stipitate, gregarious, 0.2-0.9 mm tall. Sporotheca globose to

subglobose, 0.1-0.3 mm diam., whitish with lighter zones, with a dark disc

at the base. Stipe robust, 0.1-0.5 mm tall, reddish-brown to darkish brown,

wider and darker at the base. Hypothallus membranous, discoid, concolorous

with stipe. Peridium double, with the outer membrane formed by calcareous

crosshairs, the inner membranous with iridescent blue tones. Capillitium and

columella absent. Spores globose to subglobose, black in mass, violaceous

by LM, 9-10 um diam., with a reticulate hemisphere and the opposite warty

(as confirmed by SEM; see Moreno & Mitchell 2013).

STUDIED MATERIAL: MEXICO. Sonora: Loc. 4, leg. CS, AS & AG, 7-IX-2009, obtained

from moist chamber, 4-XII-2009 (UES 8698); Loc. 8, on Lophocereus schottii, leg. AG

& AS, 21-V-2009, obtained from moist chamber, 11-VI-2009 (UES 8141, AH 42977);

obtained from moist chamber, 15-VI-2009 (UES 8142); obtained from moist chamber,

19-VI-2009 (UES 8143); obtained from moist chamber, 22-VI-2009 (UES 8144);

obtained from moist chamber, 30-VI-2009 (UES 8145); leg. CS, AS & AG, 6-XI-2009,

obtained from moist chamber, 30-XI-2009 (UES 8699); obtained from moist chamber,

11-XII-2009 (UES 8700); obtained from moist chamber, 12-XII-2009 (UES 8701);

obtained from moist chamber, 12-XII-2009 (UES 8702); Loc. 9, on cactus remains, leg.

AG & AS, 26-V-2009, obtained from moist chamber, 15-VI-2009 (UES 8146).

ComMMENTS—Didymium eremophilum is a succulenticolous species adapted to

arid environments (Blackwell & Gilbertson 1980b), characterized by its small

size, stipitate sporocarps, crystalline lime, lack of capillitium, partly reticulate

verrucose spores, and succulenticolous habitat (Moreno & Mitchell 2013).

Didymium eremophilum specimens, very difficult to find due to their small

size, have thus far been found only from moist chamber. Previously recorded in

Mexico only from Puebla (Estrada-Torres et al. 2009), our study provides a first

record for the Sonoran mycobiota.

Myxomycetes of Sonora 6 (Mexico) ... 153

Didymium squamulosum (Alb. & Schwein.) Fr.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 4, on deer manure, leg. AG & AS 12-

III-2009, obtained from moist chamber, 1-V-2009 (UES 7881); Loc. 7, on Stenocereus

thurberi, leg. AG & AS, 11-II-2009, obtained from moist chamber, 6-IV-2009 (UES

7890).

CoMMENTS— Lizarraga et al. (1999a) published a SEM study of the spore

ornamentation. Widely cited for Mexico (Moreno et al. 2007), D. squamulosum

has been previously cited for Sonora by Moreno et al. (2006) and Lizarraga et

al. (2008b).

Didymium sturgisii Hagelst. Figs 18-23

STUDIED MATERIAL: MEXICO. Sonora: Loc. 7, on Prosopis glandulosa, leg. AG & AS,

11-III-2009, obtained from moist chamber, 8-IV-2009 (UES 7884, AH 42978).

ComMMENTS—This species is characterized by its dirty white to whitish grey

flattened to subglobose fructifications (occasionally with small superficial

orifices) and sporocarp interiors with erect pillars or internally traversing

trabeculae. Moreno et al. (2001) published an SEM study of the capillitium,

pillars, and spores.

Didymium sturgisii has been recorded from Baja California (Moreno et al.

2007), Chihuahua (Salazar-Marquez et al. 2014), Puebla (Estrada-Torres et al.

2009), and Sonora (Esqueda et al. 2012, 2013).

Echinostelium apitectum K.D. Whitney

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Prosopis glandulosa leg. AS & AG,

9-III-2009, observed in moist chamber, 28-III-2009, 4-IV-2009 (UES 7892); Loc. 3, on

Stenocereus thurberi, 19-VIII-2009, obtained in moist chamber, 2-IX-2009, 5-IX-2009

(UES 8589); Loc. 4, on Olneya tesota, leg. AG & AS, 3-VI-2009, obtained from moist

chamber, 17-VI-2009 (UES 8149); Loc. 5, on decomposing wood, leg. ML, CS, AS &

AG, 18-VIII-2009, obtained in moist chamber, 2-IX-2009, 5-IX-2009 (UES 8581); Loc.

6, on Prosopis glandulosa, leg. AG & AS, 12-III-2009, observed in moist chamber, 28-

III-2009, 2-IV-2009 (UES 7902); Loc. 7, on Prosopis glandulosa, leg. AG & AS, 22-V-

2009, observed in moist chamber, 3-VI-2009, 22-VI-2009 (UES 8151); Loc. 9, Carnegiea

gigantea, leg. ML, CS, AS & AG, 20-VIII-2009, observed in moist chamber, 2-IX-2009,

7-IX-2009 (UES 8584); Loc. 10, on Prosopis glandulosa, leg. CS, AS & AG, 3-XI-2009,

observed in moist chamber, 9-XI-2009, 30-XI-2009 (UES 8703).

CoMMENTS—Previously cited from Sonora by Lizarraga et al. (2007, 2008a).

Echinostelium arboreum H.W. Keller & T.E. Brooks

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on decomposing wood, leg. AG & AS,

22-V-2009, obtained from moist chamber, 11-VI-2009 (UES 8106 with Arcyria cinerea).

154 ... Lizarraga & al.

ComMENtTS—Moreno et al. (2001) published a SEM study of the sporocarp and

spores. Previously cited from Sonora by Esqueda et al. (2013), E. arboreum has

been widely reported from Mexico (Moreno et al. 2007).

Echinostelium colliculosum K.D. Whitney & H.W. Keller

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AS & AG,

9-III-2009, 28-III-2009, obtained from moist chamber, 4-I[V-2009 (UES 7891); Loc.

5, on Guaiacum coulteri, 14-III-2009, obtained from moist chamber, 4-IV-2009 (UES

7893); on Olneya tesota, observed in moist chamber, 4-IV-2009 (UES 7894); on Prosopis

glandulosa, obtained from moist chamber, 8-IV-2009 (UES 7895); on Parkinsonia sp.,

leg. AG & AS, 26-V-2009, obtained from moist chamber, 11-VI-2009 (UES 8152); Loc.

6, on Cylindropuntia arbuscula, leg. AS & AG, 27-V-2009, 3-VI-2009, obtained from

moist chamber, 11-VI-2009 (UES 8153); on Parkinsonia sp., leg. ML, CS, AS & AG, 19-

VIII-2009, in moist chamber, 2-IX-2009, 5-IX-2009 (UES 8588); Loc. 7, on Parkinsonia

sp., leg. AS & AG, 11-III-2009, in moist chamber, 9-IV-2009 (UES 7901); on Encelia

farinosa A. Gray ex Torr., 22-V-2009, 3-VI-2009, in moist chamber, 11-V1I-2009 (UES

8150); on decomposing wood, leg. ML, CS, AS & AG, 17-VIII-2009, in moist chamber,

2-IX-2009, 5-IX-2009 (UES 8591); Loc. 9, on Guaiacum coulteri, 11-III-2009, 28-III-

2009, in moist chamber, 4-IV-2009 (UES 7896); on Olneya tesota, in moist chamber,

4-IV-2009 (UES 7898); on Cylindropuntia arbuscula, in moist chamber, 4-IV-2009 (UES

7899); leg. ML, CS, AS & AG, 20-VIII-2009, in moist chamber, 2-IX-2009, 7-IX-2009

(UES 8583); Loc. 10, on Cylindropuntia sp., leg. AS & AG, 12-III-2009, 4-IV-2009 (UES

7900); leg. ML, CS, AS & AG, 18-VIII-2009, in moist chamber, 9-IX-2009 (UES 8582);

Loc. 10, on Cylindropuntia sp., leg. AS & AG, 12-III-2009, 4-IV-2009 (UES 7900).

ComMMENTS— his species is characterized by its (60-)75-90 um high stipitate

sporocarps, globose 9-13 um diam. pale yellow spores that have a spine-like

ornamentation and articular spore wall. A LM study of sporocarps and spores

by Lizarraga et al. (2003) and SEM study by Moreno et al. (2001) have been

published.

Echinostelium colliculosum has been recorded for Baja California Sur and

Chihuahua (Moreno et al. 2007), Oaxaca and Puebla (Estrada-Torres et al.

2009), and Sonora (Esqueda et al. 2013).

Echinostelium minutum de Bary

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Prosopis glandulosa, leg. ML, CS, AS

& AG, 17-VIII-2009, in moist chamber, 2-IX-2009, 7-IX-2009 (UES 8580); Loc. 3, on

Stenocereus thurberi, leg. ML, CS, AS & AG, 17-VHI-2009, 7-IX-2009 (UES 8590); Loc. 7,

on Prosopis glandulosa, leg. AG, 22-05-2009, obtained from moist chamber, 11-06-2009

FIGURES 18-27. Didymium sturgisii (AH 42978): 18. Fructifications flattened with some calcium

carbonate granules. 19. Sporocarp (detail of small superficial orifices). 20. Capillitium and

erect pillars. 21-22. Spores (SEM). 23. Spore ornamentation (detail, SEM). Licea denudescens

(AH 42979): 24. Sporocarps. 25. Spore (SEM). Licea succulenticola (AH 42980): 26. Sporocarps.

27. Spore (SEM). Bars: 18-19 = 0.5 mm; 20 = 0.25 mm; 21-22 = 2 um; 23 = 1 um.

Myxomycetes of Sonora 6 (Mexico) ... 155

156 ... Lizarraga & al.

(UES 8154); Loc. 9, on Carnegiea gigantea, 9-I[X-2009 (UES 8585); Loc. 10, on Prosopis

glandulosa, leg. CS, AS & AG, 3-XI-2009, in moist chamber, 9-XI-2009, II-2009 (UES

8704).

ComMMENTS— Moreno et al. (2001) published a SEM study of the sporocarp

and spores. Previously reported from Sonora by Moreno et al. (2006).

Enerthenema papillatum (Pers.) Rostaf.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 4, on Forchhammeria watsonii, leg. ML,

CS, AS & AG, 19-VIII-2009, obtained in moist chamber, 5-IX-2009 (UES 8758).

CoMMENTS— Lizarraga et al. (1999a) published a SEM study of the spores from

collections from Baja California, Mexico. Widely recorded in Mexico (Moreno

et al. 2007), E. papillatus was recorded previously from Sonora by Moreno et

al. (2006).

Fuligo cinerea (Schwein.) Morgan

STUDIED MATERIAL: MEXICO. Sonora: Loc.10, on sheep manure, leg., AG, 21-V-2009,

obtained from moist chamber, 30-VI-20009 (UES 8118).

COMMENTS—Sometimes similar in size as some Badhamia species (e.g.,

B. spinispors, B. verrucospora G. Moreno et al.), Fuligo cinerea is separated by

its ovoid spores covered by small warts that join together to form a reticulum.

Cited previously for Sonora by Braun & Keller (1986) and Lizarraga et al.

(2007).

Fuligo septica (L.) EH. Wigg.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 9, on soil, leg. ML, 11-II-2009 (UES

8677).

CoMMENTS— Lizarraga et al. (1999a) published a SEM study of the spores.

Fuligo septica has a wide distribution in Mexico (Moreno et al. 2007) and has

been cited for Sonora by Pérez-Silva et al. (2001) and Lizarraga et al. (2007,

2008a,b).

Licea biforis Morgan

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AG & AS, 9-III-

2009, obtained in moist chamber, 6-IV-2009 (UES 7905); Loc. 2, on Prosopis glandulosa,

leg. AS & AG, 14-III-2009, obtained in moist chamber, 13-IV-2009 (UES 7908); Loc. 3,

on Acacia sp., 26-V-2009, obtained in moist chamber, 15-VI-2009 (UES 8159); Loc. 5,

on Guaiacum coulteri bark, leg. AS & AG 14-III-2009, obtained in moist chamber, 20-

IV-2009 (UES 7913); Loc. 6, on Prosopis glandulosa, leg. AG & AS, 27-V-2009, obtained

in moist chamber, 15-VI-2009 (UES 8162); Loc. 9, on Olneya tesota, leg. AG & AS, 11-

III-2009, obtained in moist chamber, 1-V-2009 (UES 7915); Loc. 10, leg. CS, AS & AG,

3-XI-2009, obtained in moist chamber, 12-XII-2009 (UES 8716).

Myxomycetes of Sonora 6 (Mexico) ... 157

CoMMENTS— Lizarraga et al. (1998, 1999b) published SEM studies of the

spores. Widely distributed in Mexico (Moreno et al. 2007), Licea biforis was

previously cited from Sonora by Moreno et al. (2006).

Licea denudescens H.W. Keller & T.E. Brooks Fics 24-25

Fructifications sessile, globose to subglobose, 0.1-0.3 mm diam., 0.1 mm

height, brown to blackish. Peridium membranous with some iridescent tones,

internally ornamented with small buds. Spores violaceous in mass, yellowish

brown by LM, 10-11 um diam., globose to subglobose, walls thick with pale

(thinner) areas, smooth by SEM.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AS & AG, 9-III-

2009, obtained in moist chamber, 8-IV-2009 (UES 7916, AH 42979); Loc. 5, on Olneya

tesota, leg. CS, AS & AG, 6-XI-2009, obtained in moist chamber, 12-XII-2009 (UES

8711); Loc. 7, on Parkinsonia sp., 11-III-2009, obtained in moist chamber, 20-IV-2009

(UES 7918); obtained in moist chamber, 1-V-2009 (UES 7917); Loc. 8, on Acacia greggii

A. Gray, leg. ML, CS, AS & AG, 18-VIII-2009, obtained in moist chamber, 3-X-2009

(UES 8605); Loc. 9, on cactus remains, 11-III-2009, observed in moist chamber, 15-IV-

2009 (UES 7919).

ComMMENTS—Licea denudescens is diagnosed by its thick-walled spores with

thinner areas and papillae found on the inside of the peridium. It has been

cited in Mexico from Puebla (Estrada-Torres et al. 2009), Chihuahua (Salazar-

Marquez et al. 2014), and Sonora (Esqueda et al. 2013).

Licea kleistobolus G.W. Martin

STUDIED MATERIAL: MEXICO. Sonora: Loc. 8, on Acacia greggii bark, leg. ML, CS, AS

& AG, 18-VIII-2009, observed in moist chamber, 7-IX-2009 (UES 8608).

ComMENTS—Moreno et al. (2001) published a SEM study of Licea kleistobolus,

which is characterized by its circular to subglobose fructifications with a

characteristic operculum and warted spores with groups of denser warts. Cited

previously from Sonora by Moreno et al. (2006) and Lizarraga et al. (2007).

Licea succulenticola Mosquera, Lado, Estrada & Beltran-Tej. Fics 26-27

Fructifications sessile, yellowish to dark-yellow at mature, dispersed to

gregarious, subglobose, oval to elongated, 0.1-0.3 mm long, 0.1-0.2 mm

diam., and 0.1 mm tall, with an apical and longitudinal dehiscence. Peridium

membranous, ornamented on its inner part with small warts. Spores yellow in

mass, light yellow by LM, globose to subglobose, (11-)12-14 um diam., with

a lightly paler area, with a slightly warty ornamentation. SEM examination

shows that the spore ornamentation is formed by small croziers more or less

uniformly distributed over the in spore surface.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 2, on Opuntia sp., 5-XI-2009, observed in

moist chamber, 14-XII-2009 (UES 8712); Loc. 3, on Cylindropuntia sp., leg. AS & AG,

158 ... Lizarraga & al.

14-III-2009, obtained in moist chamber, 28-III-2009, 20-IV-2009 (UES 7924); obtained

in moist chamber, 25-IV-2009 (UES 7925, 7926); Loc. 4, on Lophocereus schottii, leg.

AS & AG, 12-III-2009, obtained from moist chamber, 20-IV-2009 (UES 7923); Loc.

6, on Lophocereus schottii, leg. CS, AS & AG, 7-XI-2009, observed in moist chamber,

9-X1-2009, 9-XII-2009 (UES 8726); Loc. 7, on Lophocereus schottii, leg. AS & AG, 11-

III-2009, observed in moist chamber, 28-III-2009, 20-IV-2009 (UES 7927); obtained in

moist chamber, 1-V-2009 (UES 7928, 7929); on Stenocereus thurberi, leg. ML, CS, AS &

AG, 17-VIII-2009, observed in moist chamber, 2-IX-2009, 7-IX-2009 (UES 8747, AH

42980); obtained in moist chamber, 14-IX-2009 (UES 8602); obtained in moist chamber,

21-IX-2009 (UES 8604); obtained in moist chamber, 21-[X-2009 (UES 8603); Loc. 8, on

Lophocereus schottii, leg. ML, CS, AS & AG, 18-VIII-2009, observed in moist chamber,

2-IX-2009, 9-IX-2009 (UES 8594); obtained in moist chamber, 14-IX-2009 (UES 8595);

obtained in moist chamber, 24-IX-2009 (UES 8596); obtained in moist chamber, 30-

IX-2009 (UES 8597); obtained in moist chamber, 5-X-2009 (UES 8598, AH 42981); on

Stenocereus thurberi, leg. CS, AS & AG, 6-XI-2009, observed in moist chamber, 9 -XI-

2009, 14-XII-2009 (UES 8725); Loc. 10, on Cylindropuntia arbuscula, leg. AG & AS,

12-III-2009, obtained in moist chamber, 20-IV-2009 (UES 7920); on cactus remains,

obtained in moist chamber, 20-IV-2009 (UES 7921); obtained in moist chamber, 1-V-

2009 (UES 7922); on Lophocereus schottii, leg. ML, CS, AS & AG, 18-VIII-2009, obtained

in moist chamber, 2-IX-2009, 17-IX-2009 (UES 8599); obtained in moist chamber, 30-

IX-2009 (UES 8600); obtained in moist chamber, 30-IX-2009 (UES 8601).

ComMMENTS—Licea succulenticola resembles L. pumila G.W. Martin & R.M.

Allen (Martin & Alexopoulos 1969) in sporothecal form and spore colour.

(see Mosquera et al. 2003 for differences). Previously recorded from Hidalgo,

Morelos, Puebla, and Tlaxcala (Mosquera et al. 2003), Puebla (Estrada-Torres

et al. 2009), Sonora (Esqueda et al. 2013), and Chihuahua (Salazar-Marquez et

al. 2014).

Paradiacheopsis fimbriata (G. Lister & Cran) Hertel ex Nann.-Bremek. Fics 28-30

STUDIED MATERIAL: MEXICO. Sonora: Loc. 3, on Cylindropuntia sp., leg. AG,

14-III-2009, observed in moist chamber, 28-III-2009, 13-IV-2009 (UES 7943,

AH 42982).

COMMENTS— Widely distributed in Mexico (Moreno et al. 2007), P. fimbriata

was reported from Sonora by Moreno et al. (2006).

Perichaena depressa Lib.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on rabbit manure, leg. ML, CS, AS &

AG, 17-VIII-2009, obtained in moist chamber, 9-X-2009 (UES 8628); Loc. 9, on horse

manure, 20-VIII-2009, obtained in moist chamber, 5-X-2009 (UES 8629).

CoMMENTS— Lizarraga et al. (1998) and Moreno et al. (2001) published SEM

studies of spores and capillitium, comparing type material with samples from

Baja California and Sinaloa. Widely distributed throughout Mexico (Moreno

et al. 2007), P. depressa was previously cited for Sonora by Moreno et al. (2006).

Myxomycetes of Sonora 6 (Mexico) ... 159

FIGURES 28-36. Paradiacheopsis fimbriata (AH 42982): 28. Sporocarp. 29. Sporotheca (detail).

30. Fiber stalk (detail). Perichaena stipitata (AH 42983): 31-32. Stipitate sporocarp (detail).

33. Peridial inner surface (detail). 34. Capillitium with irregular outline. 35-36. Spores (SEM).

Bars: 28-29 = 0.1 mm; 30 = 0.02 mm; 31-32 = 0.5 mm; 33, 35-36 = 1 um; 34 = 5 um.

*Perichaena luteola (Kowalski) Gilert

STUDIED MATERIAL: MEXICO. Sonora: Loc. 10, on cow manure, leg. ML, CS, AS &

AG, 18-VIII-2009, obtained in moist chamber, 27-IX-2009 (UES 8749); 3-X-2009 (UES

8627).

ComMMENTS—The above collections have 10-13 um diam. globose to

subglobose spores that are yellow in mass (lighter yellow by LM) and with a

verruculose episporium and a yellow 2-3um diam. capillitium with smooth

surface. Lizarraga et al. (1999b) published a SEM study of material from Baja

California. Perichaena luteola is widely distributed in Mexico (Moreno et al.

2007).

160 ... Lizarraga & al.

*Perichaena stipitata Lado, Estrada & D. Wrigley Figs 31-36

Fructifications short-stipitate, solitary to gregarious, 0.3-0.8 mm tall. Stipe

robust, cylindrical, black, 0.1-0.2 mm long. Sporotheca globose to subglobose,

rarely obpyriform, yellow, 0.2-0.5 mm diam. Peridium brownish on the base,

inner surface ornamented with small rings by LM and SEM. Hypothallus dark,

membranous and irregular. Capillitium sparse, formed by yellowish branched

filaments of approximately 2 um diam., flexible, of irregular outline. Spores

yellow in mass, light yellow by LM, 12-15 um diam., globose, warted. SEM

examination shows that the spore is ornamented by warts with smooth apices

and which sometimes join into small groups.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 9, on cactus remains, leg. AG & AS, 26-

V-2209, obtained in moist chamber, 30-VI-2009 (UES 8210, AH 42983); on Lophocereus

schottii, leg. ML, CS, AS & AG, 20-VIII-2009, obtained in moist chamber, 17-IX-2009

(UES 8762); obtained in moist chamber, 24-IX-2009 (UES 8761); obtained in moist

chamber, 3-X-2009 (UES 8760, AH 42984); obtained in moist chamber, 9-X-2009 (UES

8643); Loc. 10, on Lophocereus schottii, leg. ML, CS, AS & AG, 18-VII-2009, obtained

in moist chamber, 5-X-2009 (UES 8642).

COMMENTS—Stipitate fructifications and an inner peridium ornamented by

small rings characterize this succulenticolous species. First described as new

to science by Estrada-Torres et al. (2009) from Puebla and Queretaro, Mexico,

P stipitata was later described from Arizona (USA) by Moreno & Mitchell (2013),

who published a colour plate showing its development and morphological

characteristics. A new record for Sonora.

Perichaena vermicularis (Schwein.) Rostaf.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Carnegiea gigantea, leg. CS, AS

& AG, 5-XI-2009, obtained in moist chamber, 11-XII-2009 (UES 8734); Loc. 2, on

Cylindropuntia arbuscula, leg. AS & AG, 22-V-2009, obtained in moist chamber, 25-VI-

2009 (UES 8191); Loc. 3, on Cylindropuntia sp., leg. AS & AG 14-III-2009, obtained in

moist chamber, 15-IV-2009 (UES 7953); Loc. 4, on Forchhammeria watsonii, leg. AG &

AS, 12-III-2009, obtained in moist chamber, 3-IV-2009 (UES 7944); Loc. 6, Parkinsonia

sp., 19-VIH-2009, obtained in moist chamber, 24-IX-2009 (UES 8633); Loc. 7, on

Cactaceae remains, 11-III-2009, obtained in moist chamber, 8-IV-2009 (UES 7947);

obtained in moist chamber, 20-IV-2009 (UES 7948); Loc. 8, on Cactaceae remains,

obtained in moist chamber, 18-VIII-2009, 17-IX-2009 (UES 8631).

ComMENtTs—Moreno et al. (2001) published a SEM study of spores and

capillitium in collections from Baja California that grew cespitose in moist

chamber over different substrates. Widely distributed in Mexico (Moreno et

al. 2007), P. vermicularis was previously reported from Sonora by Moreno et

al. (2006).

Myxomycetes of Sonora 6 (Mexico) ... 161

Physarum album (Bull.) Chevall.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 7, on Prosopis glandulosa, leg. AG & AS,

17-VIUI-2009, obtained in moist chamber, 9-IX-2009 (UES 8651); Loc. 10, on Prosopis

glandulosa, leg. ML, CS, AS & AG, 18-VIII-2009, obtained in moist chamber, 7-IX-2009

(UES 8650).

CoMMENTS—Lizarraga et al. (1999b) published a SEM study of spore

ornamentation in collections from Baja California. Widely distributed in

Mexico (Moreno et al. 2007) and previously reported for Sonora (Lizarraga

et al. 2007, 2008b), Physarum album frequently grows on many different

substrates when cultivated in moist chamber.

Physarum decipiens M.A. Curtis

Fructifications sessile, dispersed to grouped, subglobose or forming

short plasmodiocarp. Sporotheca yellow-greenish, 0.3-1.2 x 0.2-0.6 mm,

membranous, peridium encrusted with calcium carbonate. Capillitium

physaroid. Spores 11-13 um diam., subglobose to globose, violaceous and

slightly warty.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Prosopis glandulosa, leg. AG & AS,

9-III-2009, obtained in moist chamber, 6-IV-2009 (UES 7967); 22-V-2009, obtained in

moist chamber, 11-VI-2009 (UES 8193); Loc. 2, on Prosopis glandulosa, leg. AS & AG 14-

III-2009, obtained in moist chamber, 8-IV-2009 (UES 7970); Loc. 4, on Forchhammeria

watsonii, leg. ML, CS, AS & AG, 19-VIII-2009, obtained in moist chamber, 7-IX-2009

(UES 8649).

ComMENtTsS—Morphologically, P decipiens is highly variable. Previously cited

from Chihuahua (Lizarraga et al. 2003), Oaxaca and Puebla (Estrada-Torres et

al. 2009), Tlaxcala (Rodriguez-Palma et al. 2002), and recently Sonora (Esqueda

et al. 2012, 2013).

Physarum leucophaeum Fr.

STUDIED MATERIAL: MEXICO. Sonora: Loc. 1, on Olneya tesota, leg. AS & AG, 22-V-

2009, obtained in moist chamber, 25-VI-2009 (UES 8194); Loc. 5, on Parkinsonia sp.,

leg. AS & AG 14-III-2009, obtained in moist chamber, 1-V-2009 (UES 7966); Loc. 6, on

Encelia farinosa, 27-V-2009, obtained in moist chamber, 22-VI-2009 (UES 8198); Loc.

8, on Larrea tridentata Coville, leg. AS & AG, 21-V-2009, obtained in moist chamber,

30-VI-2009 (UES 8202); Loc. 10, on Guaiacum coulteri, leg. ML, CS, AS & AG, 18-VIII-

2009, obtained in moist chamber, 9-X-2009 (UES 8653).

ComMMENTS— Moreno et al. (2001) published a SEM study of spore

ornamentation in collections from Baja California. The recently named

species from arid regions of Eurasia and North and South America, Physarum

pseudonotabile Novozh. et al. (Novozhilov et al. 2013), is very close to P. notabile

162 ... Lizarraga & al.

T. Macbr. and P leucophaeum; distinguishing the three species is complex

and additional ecological, morphological, and molecular studies are needed.

Physarum leucophaeum is widely distributed in Mexico (Moreno et al. 2007),

common when cultured in moist chamber. Previously cited from Sonora by

Lizarraga et al. (2007, 2008b).

Acknowledgments

This research was made possible by support from the following projects: CONABIO

(Proyecto GT016). We wish to express our gratitude to Dr. S.L. Stephenson (University

of Arkansas, U.S.A.) and Mr. D.W. Mitchell (Upper Hartfield, U.K.) for reviewing the

manuscript and providing useful comments. We also extend our thanks to Mr. A. Priego

and Mr. J.A. Pérez (Electron Microscopy Service, University of Alcala de Henares)

for their invaluable help with the SEM. We also thank Luis Monje (Department of

Drawing and Scientific Photography, University of Alcala de Henares) for his help in

digitally preparing the photographs, and we are grateful to Dr. J. Rejos, curator of the

AH herbarium for his assistance with the specimens examined in the present study.

M. Lizarraga extends his gratitude to Dr. Alejandro Martinez Martinez (Jefe del Dpto.

de Ciencias Quimico Bioldgicas, ICB-UACJ) for his assistance in obtaining financial

support. C. Salazar thanks CONACYT for her scholarship to obtain a master’s degree.

Literature Cited

Andrade-Bezerra MF, Teofilo da Silva WM, Cavalcanti LH. 2008. Coprophilous myxomycetes of

Brazil: first report. Revista Mexicana de Micologia 27: 29-37.

Benjamin RK, Poitras AW. 1950. An addition to the myxomycetes genus Comatricha. Mycotaxon

42: 514-518.

Blackwell M, Gilbertson RL. 1980a. Sonoran desert Myxomycetes. Mycotaxon 11:139-149.

Blackwell M, Gilbertson RL. 1980b. Didymium eremophilum: a new myxomycete from the Sonoran

Desert. Mycologia 72: 791-797. http://dx.doi.org/10.2307/3759772

Blackwell M, Gilbertson RL. 1984. Distribution and sporulation phenology of myxomycetes in the

Sonoran desert of Arizona. Microbial Ecology 10: 369-377.

http://dx.doi.org/10.1007/BF02015561

Braun KL, Keller HW. 1986. Myxomycetes of Mexico III. Revista Mexicana de Micologia 2: 25-39.

Castillo A, Illana C, Moreno G. 1996. Badhamia melanospora Speg. A species wrongly interpreted.

Mycotaxon 57: 163-170.

Esqueda M, Lizarraga M, Gutiérrez A, Coronado ML, Valenzuela R, Raymundo T, Chacon §,

Vargas G, Barredo-Pool F. 2012. Diversidad fungica en planicies del desierto central Sonorense

y centro del desierto Chihuahuense. CIAD-UACJ-CESUES-IPN-INECOL-CICY. Informe

Final SNIB-CONABIO Proyecto GT016. México, D.F.

Esqueda M, Coronado ML, Gutiérrez A, Lizarraga M, Raymundo T, Valenzuela R. 2013. Hongos

de Reserva de la Biosfera El Pinacate y Gran Desierto de Altar. Centro de Investigacion en

Alimentacion y Desarrollo, A.C. México. 106 p.

Estrada-Torres A, Wrigley De Basanta D, Conde E, Lado C. 2009. Myxomycetes associated with

dryland ecosystems of the Tehuacan-Cuicatlan Valley Biosphere Reserve, Mexico. Fungal

Diversity 36: 17-56.

Evenson AE. 1961. A preliminary report of the Myxomycetes of Southern Arizona. Mycologia 53:

137-144. http://dx.doi.org/10.2307/3756232

Myxomycetes of Sonora 6 (Mexico) ... 163

Hernandez HM. 2006. La vida en los desiertos mexicanos. Fondo de la Cultura Econdémica,

México, DF. 188 p.

Lizarraga M, Moreno G, Illana C. 1997. The Myxomycetes from Baja California Mexico). I.

Mycotaxon 63: 287-300.

Lizarraga M, Illana C, Moreno G. 1998. First records of Myxomycetes in the state of Sinaloa, Mexico.

Micologia e Vegetazione Mediterranea 13: 167-176.

Lizarraga M, Illana C, Moreno G. 1999a. SEM studies of the Myxomycetes from the Peninsula of

Baja California Mexico), I. Arcyria to Fuligo. Annales Botanici Fennici 35: 287-306.

Lizarraga M, Illana C, Moreno G. 1999b. SEM studies of the Myxomycetes from the Peninsula of

Baja California Mexico), II. Hemitrichia to Trichia. Annales Botanici Fennici 36: 187-210.

Lizarraga M, Moreno G, Singer H, Ilana C. 2003. Myxomycetes from Chihuahua, México.

Mycotaxon 88: 409-424.

Lizarraga M, Moreno G, Illana C, Singer H. 2005. Myxomycetes from Chihuahua, Mexico III.

Mycotaxon 93: 75-88.

Lizarraga M, Moreno G, Esqueda M, Sanchez A, Coronado M. 2007. Myxomycetes from Sonora

Mexico. 3: National Forest Reserve and Wildlife Refuge Ajos-Bavispe. Mycotaxon 99: 291-301.

Lizarraga M, Moreno G, Esqueda M, Coronado M. 2008a. Myxomycetes of Sonora, México. 4:

Sierra de Alamos-Rio Cuchujaqui Biosphere Reserve. Mycotaxon. 103: 153-170.

Lizarraga M, Moreno G, Esqueda M, Coronado M. 2008b. Myxomycetes of Sonora, México. 5: Ajos-

Bavispe National Forest Reserve and Wildlife Refuge and Sierra de Alamos-Rio Cuchujaqui

Biosphere Reserve. Mycotaxon 104: 423-443.

Mandell QA, Blackwell M. 2008. Rare or rarely collected? Comatricha mirabilis from the desert of

Bahrain. Mycologia 100(5): 742-745. http://dx.doi.org/10.3852/07-154

Martin GW, Alexopoulos CJ. 1969. The Myxomycetes. University of Iowa Press. 561 pp.

Moreno G, Mitchell DW. 2013. Three rare species of Myxomycetes from Arizona and New Mexico,

USA. Boletin Sociedad Micolégica de Madrid 37: 77-83.

Moreno G, Oltra M. 2010. Notes on the genus Badhamia, Badhamiopsis and Craterium

(Myxomycetes) in Spain. Boletin Sociedad Micoldgica de Madrid 34: 161-197.

Moreno G, IIlana C, Lizarraga M. 2001. SEM studies of the Myxomycetes from the Peninsula of Baja

California (Mexico), III. Additions. Annales Botanici Fennici 38: 225-247.

Moreno G, Illana C, Esqueda M, Castillo A, Pérez-Silva E. 2004. Notes on Myxomycetes from

México. II: Boletin Sociedad Micoldégica de Madrid 28. 55-63.

Moreno G, Lizarraga M, Esqueda M, Pérez-Silva E, Herrera T. 2006. Myxomycetes de Sonora

México. II: Reserva Forestal Nacional y Refugio de Fauna Silvestre Ajos-Bavispe. Revista

Mexicana de Micologia 22: 13-23.

Moreno G, Lizarraga M, Illana C. 2007. Catalogo de los Myxomycetes de México. Boletin Sociedad

Micoldgica de Madrid 31: 187-229.

Moreno G, Mitchell DW, Novozhilov YK. 2011. A new species of Badhamia (Myxomycetes)

confused with other species with similar morphology. Boletin Sociedad Micoldgica de Madrid

35395. 102:

Mosquera J, Lado C, Estrada-Torres A, Beltran-Tejera E, Wrigley De Basanta D. 2003. Description

and culture of a new myxomycete, Licea succulenticola. Anales Jardin Botanico de Madrid

60(1): 3-10.

Novozhilov YK, Okun MV, Erastova DA, Shchepin ON, Zemlyanskaya IV, Garcia-Carvajal E,

Schnittler M. 2013. Description, culture and phylogenetic position of a new xerotolerant

species of Physarum. Mycologia 105(6): 1535-1546. http://dx.doi.org/10.3852/12-284

164 ... Lizarraga & al.

Pérez-Silva E, Herrera T, Esqueda M, Illana C, Moreno G. 2001. Myxomycetes de Sonora. I.

Mycotaxon 77: 181-192.

Poulain M, Meyer M, Bozonnet J. 2011. Les myxomycétes tomes 1-2. Fédération mycologique et

botanique Dauphiné-Savoie, Sevrier, France.

Rodriguez-Palma M, Varela-Garcia A, Lado C. 2002. Corticolous Myxomycetes associated with

four tree species in México. Mycotaxon 81: 345-355.

Salazar-Marquez C, Lizarraga M, Moreno G. 2014. Myxomycetes de matorral xer6filo del municipio

de Juarez, Chihuahua, México. Boletin Sociedad Micolégica de Madrid 38: 67-77.

Schnittler M, Novozhilov YK, Carvajal E, Spiegel FW. 2013. Myxomycete diversity in the

Tarim basin and eastern Tian-Shan, Xinjiang Prov., China. Fungal Diversity 59: 91-108.

http://dx.doi.org/10.1007/s13225-012-0186-5

MY COTAXON

http://dx.doi.org/10.5248/130.165

Volume 130, pp. 165-171 January-March 2015

New taxa of Ambomucor (Mucorales, Mucoromycotina)

from China

XIAO- YONG Lru* & RU- YONG ZHENG

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing

100101, China

* CORRESPONDENCE TO: liuxiaoyong@im.ac.cn

ABSTRACT — A new species and a new variety, A. ovalisporus and A. seriatoinflatus var.

longior, are described from soils in northwest China. A key to all known taxa of Ambomucor

accompanies descriptions and line drawings of the two new taxa. The maximum growth

temperature used as an auxiliary criterion in classification of Mucorales is also discussed.

KEY worps — morphology, taxonomy, Mucoraceae, Zygomycota

Introduction

The genus Ambomucor (Mucorales), typified by A. seriatoinflatus R.Y.

Zheng & X.Y. Liu, was introduced recently (Zheng & Liu 2014), together

with A. seriatoinflatus var. brevior R.Y. Zheng & X.Y. Liu and A. clavatus

R.Y. Zheng & X.Y. Liu. These taxa were found in northwest China. Recently,

two new Ambomucor taxa were found in the northwest regions (Tibet and

Shaanxi Province) of the country and are described here as A. ovalisporus

and A. seriatoinflatus var. longior. All Ambomucor species and varieties are

characterized by simultaneously having two kinds of sporangia: 1) fertile

sporangia containing a columella and sporangiospores and 2) aborted

sporangia that lack spores and a columella and which proliferate in chains. The

criteria for differentiating the taxa of the enlarged Ambomucor are almost the

same as previously reported (Zheng & Liu 2014). A key to all Ambomucor taxa

is presented.

Materials & methods

Isolation

A small piece of moss, humus, or dung was soaked and stirred with sterilized distilled

water. Different dilutions were poured with PDA media to make plates containing the

166 ... Liu & Zheng

antibiotic streptomycin. The isolation plates were cultivated in the basement (5-10°C,

dark) and at room temperature (about 20°C, natural solar day/night cycle). When

colonies were visible under stereomicroscopes, single colonies were transferred to PDA

slant tubes for subsequent purification and identification.

Cultures

Vouchers of the two strains studied are preserved in culture collections and the

herbarium of the Institute of Microbiology, Chinese Academy of Sciences, Beijing,

China: living cultures in the Culture Collection of the State Key Laboratory of Mycology

(Am-) and the China General Microbiological Culture Collection Center (CGMCC);

and dried cultures in the Herbarium Mycologicum Academiae Sinicae (HMAS).

Media & cultivation

PDA (pH 7) and modified SMA (dextrose 20 g, asparagine 2 g, KH,PO, 0.5 g,

MgSO,°7H,O 0.25 g, thiamine chloride 0.5 mg, agar 20 g in 1000 mL distilled water,

pH 7; Hesseltine & Ellis 1973) were used for morphological studies. PDA was also used

for the mating experiments (with 3% lecithin added) and the detection of maximum

growth temperatures. Cultivation durations and temperatures were: 1) 5-7(-10) days

at 18°C for describing morphological features; 2) 4-7 days at 25-35°C for determining

maximum growth temperatures; and 3) 7-20 days at 18°C for observing sexual reactions.

Seven-day cultures in test tubes were used to measure the height of colonies.

Taxonomy

Ambomucor ovalisporus X.Y. Liu & R.Y. Zheng, sp. nov. FIG. 1

MycoBank MB518427

Differs from all other Ambomucor spp. by its higher colonies, its wider hyphae, its fewer

sporangiophore types, its pyriform to obovoid apophysate fertile sporangia, and its

globose to oval non-vacuolate sporangiospores.

Type: China, Tibet, Nyingchi, Bomé, on wild animal dung, 17 July 2004, Xiao-yong Liu

(Holotype, HMAS 98956-1; isotypes, HMAS 98956-2,-3,-4,-5,-6,-7,-8; ex-type cultures,

Am-14, CGMCC 3.06806).

EtryMo_oey: ovalisporus, referring to the oval shape of the sporangiospores.

CoLonigs on SMA and PDA at first granulate and then floccose, 9 cm diameter

in 7 days at 18°C, up to 2.5 cm high, at first dirty white and then ‘Pinkish

Buff’ (Ridgway XXIX), reverse at first dirty white and then ‘Pinkish Cinnamon’

(Ridgway XXIX). HyPHAE 21.5-66.5(-80.5) um diameter, remaining equal

or narrowed at septa, hyaline to yellow-brown. STOLONs absent. RHIZOIDS

scanty, arising from various parts of the sporangiophores and from fertile and

aborted sporangia. SPORANGIOPHORES erect or recumbent, arising directly

from substrate or hyphae, forming both fertile and aborted sporangia always

in a sympodial pattern, (37-)322-1416(-3476) um long, 5.5-79 um diameter,

usually equal or subequal in width throughout, always constricted just beneath

the terminal fertile sporangia, hyaline to brown, with or without granular

contents, always verrucate; main axes mostly simple, single, rarely with 2-3

Ambomucor sp. & var. nov. (China) ... 167

FiGurE 1. Ambomucor ovalisporus (CGMCC 3.6806). a, Sporangiophore apex triple branches

with an aborted sporangium at the point of separation; b, Single sporangiophore with a fertile

top sporangium (in black); c-d, Rhizoids; e-f, Aborted sporangia showing different sizes;

g-i, Columellae with distinct collars; j, Sporangiospores. Scale bars: a-f = 100 um; g-i = 25 um;

j= 10um.

verticillate branches. SeEpTA appearing frequently in the hyphae and the

main axes of sporangiophores, sporadically in the sporangiophore branches.

ABORTED SPORANGIA proliferating repeatedly on the sides of sporangiophores,

up to 30 or more, transferring their contents forward and forming a short

or long chain of aborted sporangia and terminating in a normal fertile

sporangium, pyriform to obovoid and 20-67.5(-83) x 16-58.5(-—76) um, or

globose to subglobose and (12.5-)24-85 um diameter, verrucate, subhyaline.

FERTILE SPORANGIA borne terminally on the main axes or the branches of

sporangiophores, apophysate, thin-walled, verrucate, globose to subglobose,

168 ... Liu & Zheng

26-163.5 um diameter, dark brown when mature, breaking in larger ones,

persistent in smaller ones. COLUMELLAE forming in the fertile sporangia

only, well developed, verrucate, with a distinct collar, very regular in shape,

hyaline; larger ones subglobose and 102.5-186.5 um diameter, or oblate and

75.5-92(-179) x 55-73.5(-122) um; smaller ones applanate and 34.5-39.5 x

26.5 um. SPORANGIOSPORES forming in fertile sporangia only, released through

breaking or moving backward to the sporangiophores, smooth, mostly oval and

6-12.5 x 4.5-9 um, sometimes globose or subglobose and (5.5-)6.5-9(-10) um

diameter, hyaline. CHLAMYDOSPORES absent. ZYGOSPORES unknown.

ComMENTs: The maximum growth temperature for Ambomucor ovalisporus

Am-14 was determined as 27°C (results of two tests). Crosses of Ambomucor

ovalisporus with A. clavatus and A. seriatoinflatus var. seriatoinflatus, var.

brevior, and var. longior all failed to produce zygospores.

Ambomucor seriatoinflatus var. longior X.Y. Liu & R.Y. Zheng, var. nov.

MycoBank MB518428 Fic. 2

Differs from other Ambomucor seriatoinflatus varieties by its higher colonies, its

abundant aerial hyphae, its longer narrower sporangiophore branches, and its later

sporulation.

Type: China, Shaanxi Province, Taibai Mountain, Shangbansi Temple, in moss and

humus on rock, 11 October 2002, Xue-wei Wang (Holotype, HMAS240174-1; isotypes,

HMAS 240174-2,-3,-4,-5,-6,-7,-8; ex-type cultures, Am-16, CGMCC 3.014179).

Erymo_oey: longior, referring to the sporangiophore primary branches being longer

than those of Ambomucor seriatoinflatus var. seriatoinflatus.

Cotonizs on SMA and PDA floccose, zonate after 3 days, reaching 9 cm

diameter in 6-7 days at 18°C, about 3-5 mm high, at first white, then grayish

white, at last grayish white on the edge and ‘Dark Purplish Gray’ (Ridgway LHI)

in the middle, reverse ‘Olive-Buff’ (Ridgway XL). HypHaz branched, nonseptate

when young, septate in age, 3-7(-8) um diameter in substrate, abundant in

air and (1-)1.5-5(-8) um diameter. SroLons absent. RHIZOIDs scanty, not

opposite sporangiophores, arising from various parts of the sporangiophores

or from the aborted sporangia. SPORANGIOPHORES erect or recumbent, arising

directly from substrate or aerial hyphae, forming fertile or aborted sporangia, of

three main types: 1) bearing normal fertile sporangia only, 2) aborted sporangia

only, or 3) both simultaneously; main axes of the sporangiophores rarely

simple, usually branched, 2.9-7.5 um diameter from substrate, 2-3(-3.5) um

diameter from aerial hyphae; primary branches 1-3, single, in pairs, more often

monopodially or pseudoverticillately rebranching, sometimes sympodially and

branching no more than three times, (15-)60-543(-1600) um long, 2-8.5 um

diameter; main axes and branches more or less straight, or subcurved, usually

equal or subequal in width throughout, hyaline to pale greenish brown, with

Ambomucor sp. & var. nov. (China) ... 169

FiGuRE 2. Ambomucor seriatoinflatus var. longior (CGMCC 3.14179). a-e, Upper portions of

sporangiophores showing various structures: aborted sporangia (blank) and fertile sporangia

(shadowed); f, Rhizoids; g-h, Chlamydospores; i-q, Columellae with distinct collars;

r, Sporangiospores. Scale bars: a~h = 100 um; i-q = 25 um; r= 10 um.

or without granular contents, becoming vacuolate in age. SEPTA in main axes

and branches of the sporangiophores, usually present at the place of branching,

sometimes without definite position. FERTILE SPORANGIA appearing after three

days, borne terminally on the main axes or branches of the sporangiophores,

globose to slightly depressed globose, (21.5-)24.5-47(-53) um diameter in

larger ones on the sporangiophores from substrate, 7-17 um diameter in smaller

ones on the sporangiophores from aerial hyphae, non-apophysate, breaking or

slowly dissolving, thin-walled, dark brown when mature, pale grayish brown

in broken pieces of wall. ABORTED SPORANGIA borne terminally on the main

170 ... Liu & Zheng

axes and branches of sporangiophores, or repeatedly proliferating terminally or

laterally, forming intercalary sporangiophores between two aborted sporangia,

no more than three times, transferring their contents forward until a fertile and

normal sporangium is formed, non-apophysate or slightly apophysate, ovoid

and 10-24 x 7-19 um, globose and 13-22(-26) um diameter, granulate when

young, vacuolate in age, subhyaline. COLUMELLAE formed in fertile sporangia

only, well developed, very regular in shape, ovoid to ellipsoid-ovoid, depressed-

globose to sub-applanate, 6.5-21 x 5-16.5 um, hyaline, some with grayish to

brownish content, smooth, regularly with a distinct collar. SPORANGIOSPORES

forming in fertile sporangia, ellipsoid to broadly oblong-ellipsoid and 3.5-7 x

2-3.5 um, (1-)2-guttulate, hyaline, grayish in mass, smooth, becoming vacuolate

in old cultures. CHLAMYDOSPORES rare, single, or in short chains, subglobose,

ovoid or irregular, 5-18.5 x 3-14 um, hyaline to yellow. ZyGosporEs unknown.

ComMENTs: The maximum growth temperature for Ambomucor seriatoinflatus

var. longior Am-16 was determined as 30°C (results of two tests). Crosses of

Ambomucor seriatoinflatus var. longior with A. clavatus, A. ovalisporus, and

A. seriatoinflatus var. seriatoinflatus and var. brevior all failed to produce

zygospores.

Key to the taxa of Ambomucor

(revised from Zheng & Liu 2014)

1. Colonies up to 25 mm high; hyphae 21.5-66.5 um diameter; both fertile and aborted

sporangia borne on the same sporangiophore; fertile sporangia apophysate;

sporangiospores globose to oval, avacuolate; maximum growth temperature

Dp A stag shy bo tueee cbse shag Er Sabah ht hal bh Ptah obs Fb hy Sea he ral ob hy Pom okt Aa A. ovalisporus

1. Colonies 1-5 mm high; hyphae 1.5-14 «um diameter; fertile or aborted sporangia

or both borne on the same sporangiophore; fertile sporangia non-apophysate;

sporangiospores ellipsoid to oblong-ellipsoid, vacuolate; maximum growth

temperature no less than B0°C ens 5 te ss Ue ae stews a hw saws OE a ee oe Z

2. Aborted sporangia oval to ellipsoidal; columellae of the fertile sporangia mostly

applanate, rarely globose to subglobose; aborted outgrowths frequent, often

spirally twisted; maximum growth temperature 33°C ............. A. clavatus

2. Aborted sporangia globose, subglobose to ovoid; columellae of the fertile sporangia

usually not as above; aborted outgrowths absent or rare, not spirally twisted

when present; maximum growth temperature 30°C ...................0008 3

3. Aborted sporangia borne in chains of up to 20; aborted outgrowths usually absent

and not in a zigzag manner; main axes and branches more or less straight, rarely

SHOE VACUEVED 1 tp. Bess Nak vaso kmh skies 2 A. seriatoinflatus var. seriatoinflatus

3. Aborted sporangia usually not observed exceeding chains of 6; aborted outgrowths

when present formed sympodially; main axes and branches subcurved and

fately: straieht ta: Sub-Strar int! Far 55: Face nas: Bape Pape oP aps eB Pees Pee BSc le Bate 4

Ambomucor sp. & var. nov. (China) ... 171

4. Colonies 3-5 mm high; aerial hyphae abundant; fertile sporangia appearing after

three days; aborted sporangia globose when diameter is less than 26 um;

columellae less than 21 um in width ............ A. seriatoinflatus var. longior

4. Colonies 1-3 mm high; aerial hyphae scarce; fertile sporangia appearing after one

day; aborted sporangia globose, up to 66 um in diameter; columellae 42 um or

TOTS WIGS Pe te coal ei cu Bea teva Mh eva Men td tn At own A A. seriatoinflatus var. brevior

Discussion

Based on the results of studies on several genera of Mucorales, such as

Actinomucor (Zheng & Liu 2005), Cunninghamella (Zheng & Chen 2001),

Pilaira (Zheng & Liu 2009), Rhizomucor (Zheng et al. 2009), and Rhizopus

(Zheng et al. 2007), the range of maximum growth temperatures (MGT) is an

important criterion for helping circumscribe genera and distinguish species;

however, it is not a decisive characteristic and sometimes there is overlap

among taxa. The MGTs of the three Ambomucor species (A. ovalisporus 27°C;

A. seriatoinflatus 30°C; A. clavatus 33°C) occupy a narrow range that supports

their generic afhnity, but with significant differences that support the species

delimitation.

Acknowledgments

The study was partially supported by the National Natural Science Foundation of

China (Nos. 31070019 and 31370068), the Ministry of Science and Technology of China

(No. 2012FY111600) and the Foundation of the Knowledge Innovation Program of the

Chinese Academy of Sciences (No. KSCX2-EW-J-6). We greatly appreciate Associate Prof.

Xue-wei Wang of this department for collecting and isolating the type strain of Ambomucor

seriatoinflatus var. longior. We also thank Ms. Hong-mei Liu of this research group for

preparing the media and plates, helping with isolation, doing mating experiments,

and testing the maximum growth temperatures. Finally we thank Dr. Gerald L. Benny

(University of Florida, Gainesville, U.S.A.) and Dr. Bo Huang (Anhui Agricultural

University, Hefei, P.R. China) for critical comments and presubmission review.

Literature cited

Hesseltine CW, Ellis JJ. 1973. Mucorales. 187-217, in: GC Ainsworth, et al. (eds). The fungi, vol. 4b.

Academic Press, New York.

Zheng RY, Chen GQ. 2001. A monograph of Cunninghamella. Mycotaxon 80: 1-75.

Zheng RY, Liu XY. 2005. Actinomucor elegans var. meitauzae, the correct name for A. taiwanensis

and Mucor meitauzae (Mucorales, Zygomycota). Nova Hedwigia 80: 419-431.

http://dx.doi.org/10.1127/0029-5035/2005/0080-0419

Zheng RY, Liu XY. 2009. Taxa of Pilaira (Mucorales, Zygomycota) from China. Nova Hedwigia 88:

255-267. http://dx.doi.org/10.1127/0029-5035/2009/0088-0255

Zheng RY, Liu XY. 2014 [“2013”]. Ambomucor gen. & spp. nov. from China. Mycotaxon 126:

97-108. http://dx.doi.org/10.5248/126.97

Zheng RY, Chen GQ, Huang H, Liu XY. 2007. A monograph of Rhizopus. Sydowia 59: 273-372.

Zheng RY, Liu XY, Li RY. 2009. More Rhizomucor causing human mucormycosis from China:

R. chlamydosporus sp. nov. Sydowia 61: 135-147.

MY COTAXON

http://dx.doi.org/10.5248/130.173

Volume 130, pp. 173-179 January-March 2015

Perenniporia koreana, anew wood-rotting basidiomycete

from South Korea

YEONGSEON JANG’, SEOKYOON JANG’, YOUNG WOON LIM’,

CHANGMU KIM? & JAE-JIN K1m’*

‘Division of Environmental Science & Ecological Engineering,

College of Life Sciences & Biotechnology, Korea University,

5-1 Anam-dong, Seongbuk-gu, Seoul, 136-701, Korea

School of Biological Sciences, Seoul National University, Seoul, 151-747, Korea

°National Institute of Biological Resources, Environmental Research Complex,

Incheon, 404-708, Korea

*CORRESPONDENCE TO: jae-jinkim@korea.ac.kr

ABSTRACT — A new Perenniporia species, P. koreana, is described based on morphological

and molecular data from specimens collected from Gyeonggi-do and Gangwon-do, Korea.

It is characterized by annual resupinate basidiocarps that are very pale brown to yellow

when dry. Microscopically, it has a dimitic hyphal system with dextrinoid and cyanophilous

skeletal hyphae, and its basidiospores are ellipsoid, hyaline, thick-walled, smooth, dextrinoid,

cyanophilous, (5.9-)6-7(-7.5) x (3.7-)3.9-5.2(- 5.7) um.

Key worps —phylogeny, Polyporaceae, polypore, taxonomy

Introduction

Perenniporia Murrill is a polypore genus of wood-rotting fungi. The

characteristics of this genus are: (1) annual to perennial, resupinate to pileate

basidiocarps; (2) dimitic to trimitic hyphal system with clamped generative

hyphae and non-dextrinoid to strongly dextrinoid or amyloid vegetative

hyphae; and (3) usually thick-walled, ellipsoid to truncate, variably dextrinoid

or amyloid basidiospores (Gilbertson & Ryvarden 1987; Zhao & Cui 2013a,b).

There are about 90 Perenniporia species reported in the world and this genus

was shown to be polyphyletic when evolutionary relationships were inferred

from the representative Perenniporia species including the type, P. medulla-

panis (Jacq.) Donk (Robledo et al. 2009; Zhao et al. 2013; Zhao & Cui 2013a,b).

174 ... Jang & al.

In Korea, nine Perenniporia species have been recorded: P. fraxinea (Bull.)

Ryvarden, P. fraxinophila (Peck) Ryvarden, P. medulla-panis, P. minutissima

(Yasuda) T. Hatt. & Ryvarden, P. ochroleuca (Berk.) Ryvarden, P. ohiensis (Berk.)

Ryvarden, P. subacida (Peck) Donk, P. tephropora (Mont.) Ryvarden, and P

truncatospora (Lloyd) Ryvarden (Lee & Jung 2005; Park & Lee 2011). During

research on indigenous fungi in South Korea, we found eight specimens that

represent Perenniporia but had characteristics that deviated from the previously

reported species. Based on morphological and molecular analyses, we describe

them as a new species.

Materials & methods

Collection and morphological examination

Eight Perenniporia specimens collected from Gyeonggi-do and Gangwon-do

in Korea were used in this study. We examined them macro- and microscopically

according to Jang et al. (2013). Spores were measured in Melzer’s reagent and 5% of

the measurements from both ends are presented in parentheses (Zhao & Cui 2013a).

Color codes follow Munsell (2009). The studied specimens have been deposited at the

Herbarium of National Institute of Biological Resources, Korea (KB).

Phylogenetic analysis

Genomic DNA was extracted from the specimens according to Jang et al. (2012).

PCR reactions were primed with ITSIF (Gardes & Bruns 1993) / ITS4 (White et al.

1990) for internal transcribed spacer (ITS) region and with LROR/LR7 (Vilgalys &

Hester 1990) for nuclear large subunit ribosomal DNA (nLSU) region according to Jang

et al. (2012). DNA was sequenced by Macrogen DNA sequencing service (Seoul, Korea),

and the sequences were deposited in GenBank, NCBI. The ITS and nLSU sequences of

closely related species were retrieved from GenBank. The sequences were aligned with

MAFFT 7.130 (Katoh & Standley 2013) using the L-INS-i method for each dataset. For

Bayesian analysis, the best-fit model for each dataset was calculated by MrModeltest 2.3

(Nylander 2004) under AIC criterion. The two datasets were combined and the selected

models were applied. The 50% majority rule consensus tree was calculated by MrBayes

3.2 (Ronquist et al. 2012) according to Jang et al. (2013).

Taxonomy

Perenniporia koreana Y. Jang & J.J. Kim, sp. nov. Fic. 1

MycoBank MB 807626

Differs from other Perenniporia species by its annual resupinate basidiocarps with grayish

orange pore surface, lack of rhizomorphs in most specimens, a dimitic hyphal system

with dextrinoid skeletal hyphae, and ellipsoid non-truncate dextrinoid basidiospores.

Fic. 1. Perenniporia koreana (KB NIBRFG107080, holotype). A. Basidiocarp. Scale bar = 1 cm.

B. Microscopic features: a, basidioles and basidia; b, cystidioles; c, basidiospores; d, generative

hyphae from trama; e, generative hyphae from context; f, skeletal hyphae from trama; g, skeletal

hyphae from context.

koreana sp. nov. (South Korea) ... 175

iporia

Perenn

176 ... Jang & al.

Type: Korea. Kyeonggi-do, Dongducheon-si, Mt. Soyo, 37°56’44”N 127°04’08’E, on

the branch of Quercus sp., 17 May 2008, Jae-Jin Kim KUC20080517-15 (Holotype, KB

NIBRFG107080; GenBank KJ156301, KJ156309).

Erymo.oey: The locality (Korea) of the type specimen.

BASIDIOCARPS annual, resupinate, adnate, ca. 8.5 cm or more in longest

dimension, ca. 4 cm in widest dimension. Sterile margin felty, orange gray

(10YR8/2), up to 1 cm wide, usually without rhizomorphs (present only

in KB NIBRFG107396). Pores round to angular, 5-6 per mm; dissepiments

thin, entire; pore surface grayish orange (1OYR8/3-8/4) to brownish orange

(10YR7/6-7/8) in dried condition. Context corky, cream to buff, up to 0.5 mm

thick. Tubes corky, concolorous with pore surface, up to 2 mm thick.

HyPHAL SYSTEM dimitic; generative hyphae with clamp connections; skeletal

hyphae dextrinoid, cyanophilous.

CONTEXT generative hyphae hyaline, thin-walled, 1.5-3 um in diameter;

skeletal hyphae dominant, hyaline, thick-walled, with a distinct lumen,

interwoven, 3.5-6 um in diameter.

TUBES generative hyphae hyaline, thin-walled, 1.5-2.5 um in diameter;

skeletal hyphae dominant, hyaline, thick-walled, with a narrow lumen to

subsolid, rarely branched, interwoven, 1.5-4.5 um in diameter. Cystidia

absent, fusoid cystidioles present, 19.5-23 x 5-6.5 um; basidia barrel-shaped,

4 sterigmate, 21-23 x 7.5-9 um.

Basiprospores ellipsoid, hyaline, thick-walled, smooth, dextrinoid,

cyanophilous, (5.9-)6-7(-7.5) x (3.7-)3.9-5.2(-5.7) um, L = 6.56, W = 4.45

(n = 120/4).

ECOLOGY & DISTRIBUTION — Korea, on hardwoods causing white rot.

ADDITIONAL SPECIMENS EXAMINED: KOREA, KYEONGGI-DO, Uijeongbu-si, Mt.

Dobong, on fallen wood branch, 30 October, 2009, Yeongseon Jang KUC20091030-

32 (KB NIBRFG113564; GenBank KJ156305, KJ156313); Yongin-si, Mt. Gwanggyo,

37°21'05"'N 127°03’14’E, on hardwood, 14 August 2012, Yeongseon Jang KUC20120814-

17 (KB NIBRFG125415; GenBank KJ156306, KJ156314); Yangju-si, Mt. Goryeong, on

wood, 11 July 2009, Yeongseon Jang KUC20090711-70 (KB NIBRFG113613; GenBank

KJ156304, KJ156312); Dongducheon-si, Mt. Soyo, 37°56’44”N 127°04’08’E, on the

branch of hardwood, 17 May 2008, Jae-Jin Kim KUC20080517-02 (KB NIBRFG107071;

GenBank KJ156300, KJ156308); GANGWON-DO, Heongseong-gun, Mt. Chiak, on wood,

4 April 2009, Young Woon Lim KUC20090404-27 (KB NIBRFG113243; GenBank

KJ156303, KJ156311); 37°20’16”N 128°03’27”E, on hardwood, 2 October 2008, Jae-Jin

Kim KUC20081002J-02 (KB NIBRFG107396; GenBank KJ156302, KJ156310); Wonju-

si, Mt Chiak, 37°21’43”N 128°02’36’E, on the branch of Quercus sp., 10 May 2008, Jae-

Jin Kim KUC20080510-01 (KB NIBRFG107760; GenBank KJ156299, KJ156307).

Phylogeny

The combined ITS+nLSU dataset contained 56 taxa and 2016 characters, of

which the ITS dataset was 667 characters and nLSU was 1349 characters. By the

Perenniporia koreana sp. nov. (South Korea) ... 177

1 Perenniporiella chaquenia MUCL 47648 (FJ411084/FJ393856)

1 Perenniporiella pendula MUCL 46034 (FJ411081/FJ393853)

258 Perenniporiella micropora MUCL 43581 (FJ411086/FJ393858)

0.96 Perenniporia minor Cui 5782 (HQ883475/HQ6541 15)

1p Perenniporia lacerata Cui 7220 (JX141448/JX141458)

Perenniporia macropora Zhou 407 (JQ861746/JQ861 762)

968 Perenniporia tibetica Cui 9457 (JF706326/JF706332)

Perenniporia nanlingensis Cui 7620 (HQ848477/HQ848486)

0.97 Perenniporia japonica Cui 7047 (HQ654097/HQ6541 11)

0.79 Perenniporia ochroleuca Dai 11486 (HQ654105/JF706349)

0.82 4 Perenniporia detrita MUCL 42649 (FJ411099/FJ393866)

0.94 Perenniporia ohiensis Cui 5714 (HQ654103/HQ654116)

1 Abundisporus sclerosetosus MUCL 41438 (FJ411101/FJ393868)

Abundisporus violaceus MUCL 38617 (FJ411100/FJ393867) |

1 Perenniporia substraminea Cui 10177 (JQ001852/JQ001 844)

0.66 4 Perenniporia medulla-panis MUCL 43520 (FJ411087/FJ393875)

O87 Perenniporia hainaniana Cui 6364 (JQ861743/JQ861759)

Perenniporia straminea Cui 8718 (HQ876600/JF 706335)

222 093 Perenniporia piceicola Dai 4184 (JF706328/JF 706336)

Perenniporia russeimarginata Yuan 1225 (JQ861749/JQ861 765)

1p Perenniporia truncatospora Cui 6987 (JN048778/HQ6541 12)

Perenniporia pyricola Dai 10265 (JN048761/JN048781)

0.51 9 Perenniporia tephropora Cui 6331 (HQ848473/HQ848484)

Perenniporia subtephropora Dai 10962 (JQ861752/JQ861 768)

0.99 Perenniporia aridula Dai 12396 (JQ001854/JQ001846)

Perenniporia tenuis Wei 2783 (JQ001858/JQ001848)

4 Perenniporia maackiae Cui 5605 (JN048760/JN048780)

Perenniporia corticola Dai 7330 (HQ654094/HQ654108)

Perenniporia koreana KUC20090711-70 (KJ156312, KJ156304)

°97) Perenniporia koreana KUC20080517-15 (KJ156309, KJ156301) - holotype

Perenniporia koreana KUC20080517-02 (KJ156308, KJ156300)

ip Perenniporia koreana KUC20090404-27 (KJ156311, KJ156303)

1 1] | Perenniporia koreana KUC20081002J-02 (KJ156310, KJ156302)

Perenniporia koreana KUC20120814-17 (KJ156314, KJ156306)

Perenniporia koreana KUC20091030-32 (KJ156313, KJ156305)

4 Perenniporia koreana KUC20080510-01 (KJ156307, KJ156299)

1p Perenniporia luteola Harkonen 1308b (JX141457/JX141467)

; 0.99 Perenniporia luteola Harkonen 1308a (JX141456/JX141466)

1 Perenniporia rhizomorpha Dai 7248 (JF706330/JF706348)

Perenniporia rhizomorpha Cui 7507 (HQ654107/HQ654117)

1p Perenniporia bannaensis Cui 8562 (JQ291728/JQ291 730) | |

Perenniporia bannaensis Cui 8560 (JQ291727/JQ291729)

yeh 4 Perenniporia subacida MUCL 31402 (FJ411103/FJ393880)

Perenniporia narymica Dai 10510 (HQ654101/JF 706346)

1 Perenniporia subadusta Cui 8459 (HQ876606/HQ654113)

O74 Microporellus violaceo-cinerascens MUCL 45229 (FJ411106/FJ393874)

oes Perenniporia martia MUCL 41677 (FJ411092/FJ393859)

1--Perenniporia fraxinea Cui 8871 (JF706329/JF706345)

i Perenniporia robiniophila Cui 9174 (HQ876610/JF706343)

4 Perenniporia vicina MUCL 44779 (FJ411095/FJ393862)

7 Perenniporia formosana Dai 5245 (HQ876612/JX941590)

051 Perenniporia tianmuensis Cui 2648 (JX141453/JX141463)

1 Perenniporia contraria Knudsen 04-111 (JQ861737/JQ861755)

Perenniporia fergusii Gilbertson 16116 (HQ876607/JF 706337)

1 Pyrofomes demidoffii MUCL 41034 (FJ411105/FJ393873)

Donkioporia expansa MUCL 35116 (FJ411104/FJ393872)

0.01

Fic. 2. 50% majority consensus tree of Perenniporia koreana and allied species inferred from

combined ITS+nLSU region sequences. Posterior probabilities over 0.5 are shown above branches.

GenBank accession numbers are presented in parentheses (ITS/nLSU). Perenniporia koreana is

indicated by bold type.

model selection, GTR+I+G model was chosen for both ITS and nLSU datasets.

Pyrofomes demidoffii (Lév.) Kotl. & Pouzar (MUCL 41034) and Donkioporia

expansa (Desm.) Kotl. & Pouzar (MUCL 35116) were used as outgroup (Zhao

& Cui 2013a,b; Zhao et al. 2013). The phylogenetic tree (Fic. 2) provides

the sequence information for the included taxa. Our analysis supported two

main groups, I and II. Group I, which contained Perenniporia species with

typically truncate basidiospores, included P. medulla-panis (the type species)

as well as Abundisporus sclerosetosus Decock & Laurence, A. violaceus (Wakef.)

Ryvarden, Perenniporiella chaquenia Robledo & Decock, P pendula Decock &

Ryvarden, and P. micropora (Ryvarden) Decock & Ryvarden. Group IJ included

178 ... Jang & al.

Perenniporia species with non-truncate basidiospores (except for P fergusii

Gilb. & Ryvarden with ellipsoid to slightly truncate basidiospores; Gilbertson

& Ryvarden 1987), and Microporellus violaceocinerascens (Petch) A. David &

Rajchenb. Perenniporia koreana was in group II and monophyletic with high

posterior probability value (1.0 p.p.). Its sister taxa were P. luteola B.K. Cui &

C.L. Zhao and P. rhizomorpha B.K. Cui et al., and these three taxa clustered

with P bannaensis B.K. Cui & C.L. Zhao, P. subacida, and P. narymica (Pilat)

Pouzar in a clade with high support (1.0 p.p.).

Discussion

Morphologically, Perenniporia koreana is very similar to its sister taxa,

P. luteola and P. rhizomorpha, which are also characterized by a resupinate

basidiocarp, yellowish pore surface, dextrinoid skeletal hyphae, and dextrinoid

non-truncate basidiospores. However, P. luteola differs by its perennial habit

and slightly wider basidiospores (6.1-6.9 x 5.1-5.4 um; Zhao & Cui 2013a),

and P. rhizomorpha differs by having typical rhizomorphs, slightly longer

basidiospores (5.3-6.5 x 4.1-5.2 um), contextual skeletal hyphae encrusted

with fine crystals and lacking cystidia and cystidioles (Cui et al. 2007).

Perenniporia tenuis (Schwein.) Ryvarden and P. bannaensis are similar to

P. koreana in their annual resupinate basidiocarps, yellowish pore surface,

dimitic hyphal system with dextrinoid skeletal hyphae, fusoid cystidioles, and

dextrinoid basidiospores. However, P. tenuis differs in its larger pores (3-5/mm)

and small ellipsoid to truncate basidiospores (4.5-6 x 3.5-4.5 um; Ryvarden

& Gilbertson 1994), while P bannaensis differs in its small pores (6-8/mm),

entire to lacerate dissepiments, and shorter basidiospores (5.2-6 x 4-4.6 um;

Zhao et al. 2013).

Phylogenetically (Fic. 2), the species relationships in our analysis differ

substantially from those of Zhao et al. (2013) and Zhao & Cui (2013a,b). Our

analysis clustered Perenniporia luteola, PR. rhizomorpha, and P. bannaensis in

group II, a relationship also observed when P. koreana was excluded from

the phylogenetic analysis (not shown). In contrast, P rhizomorpha and

P. bannaensis were monophyletic with P. medulla-panis in Zhao et al. (2013);

and P. luteola, P. rhizomorpha, and P. bannaensis clustered with P medulla-panis

in Zhao & Cui (2013a,b). Further research with additional loci would help to

resolve the species relationships of Perenniporia.

In conclusion, we propose Perenniporia koreana as a new species with only

minute morphological distinction but with distinct genetic differences.

Acknowledgments

This research was supported by Basic Science Research Program through the

National Research Foundation of Korea (NRF) funded by the Ministry of Education

(NRF-2013R1A1A2A10011390) and was supported by the Indigenous Species Survey

Perenniporia koreana sp. nov. (South Korea) ... 179

and Investigation project from the National Institute of Biological Resources (NIBR)

under the Ministry of Environment, Republic of Korea. We are much obliged to Dr.

Bao-Kai Cui and Dr. Tsutomu Hattori for their valuable suggestions on the manuscript.

Literature cited

Cui BK, Dai YC, Decock C. 2007. A new species of Perenniporia (Basidiomycota, Aphyllophorales)

from eastern China. Mycotaxon 99: 175-180.

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes — application

to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.

http:/dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Gilbertson RL, Ryvarden L. 1987. North American polypores, vol. 2. Fungiflora, Oslo.

Jang Y, Choi HE, Lim Y W, Lee JS, Kim J-J.2012. The first report of Ceriporia lacerata (Phanerochaetaceae,

Basidiomycota) in Korea. Mycotaxon 119: 397-403. http://dx.doi-org/10.5248/119.397

Jang Y, Lee SW, Lim YW, Lee JS, Hallenberg N, Kim J-J. 2013. Hypochnicium pini, a new corticioid

basidiomycete in East Asia. Mycotaxon 124: 209-217. http://dx.doi.org/10.5248/124.209

Katoh K, Standley DM. 2013. MAFFT Multiple sequence Alignment Software Version

7: Improvements in Performance and Usability. Mol. Biol. Evol. 30: 772-780.

http://dx.doi.org/10.1093/molbev/mst010

Lee JS, Jung HS. 2005. List of recorded Korean Aphyllophorales. Kor. J. Mycol. 33: 38-53.

Munsell color. 2009. Munsell soil color charts with genuine Munsell color chips. Grand Rapids,

Michigan, U.S.A

Nylander JAA. 2004. MrModeltest v2. Evolutionary Biology Center, Uppsala University, Uppsala,

Sweden.

Park W, Lee J. 2011. New wild Fungi of Korea. Kyohak Publishing, Korea.

Robledo GL, Amalfi M, Castillo G, Rajchenberg M, Decock C. 2009. Perenniporiella chaquenia

sp. nov. and further notes on Perenniporiella and its relationships with Perenniporia (Poriales,

Basidiomycota). Mycologia 101: 657-673. http://dx.doi.org/10.3852/08-040

Ronquist F, Teslenko M, Mark P van der, Ayres D, Darling A, Hohna S, Larget B, Liu L, Suchard MA,

Huelsenbeck JP. 2012. MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice

across a large model space. Syst. Biol. 61: 539-542. http://dx.doi.org/10.1093/sysbio/sys029

Ryvarden L, Gilbertson RL. 1994. European polypores 2. Synopsis fungorum 7. Fungiflora, Oslo.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified

ribosomal DNA from several Cryptococcus species. J. Bacteriol. 172: 4238-4246.

White TJ, Bruns TD, 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, New York.

Zhao CL, Cui BK. 2013a. Three new Perenniporia (Polyporales, Basidiomycota) species

from China based on morphological and molecular data. Mycoscience 54: 231-240.

http://dx.doi.org/10.1016/j.myc.2012.09.013

Zhao CL, Cui BK. 2013b. Morphological and molecular identification of four new resupinate

species of Perenniporia (Polyporales) from southern China. Mycologia 105: 945-958.

http://dx.doi.org/10.3852/12-201

Zhao CL, Cui BK, Dai YC. 2013. New species and phylogeny of Perenniporia based on morphological

and molecular characters. Fungal Divers. 58: 47-60. http://dx.doi.org/10.1007/s13225-012-0177-6

MY COTAXON

http://dx.doi.org/10.5248/130.181

Volume 130, pp. 181-190 January-March 2015

New records of corticolous myxomycetes from Turkey

R. BATUR ORAN! & C. CEM ERGUL"

Department of Biology, Faculty of Arts & Sciences, Uludag University,

16059, Bursa, Turkey

*CORRESPONDENCE TO: ergulc@uludag.edu.tr

ABSTRACT- Eight myxomycete species from the Marmara region are newly recorded for

Turkey: Amaurochaete tubulina, Comatricha longipila, Diderma cinereum, D. umbilicatum,

Didymium lenticulare, Physarum serpula, Stemonitis inconspicua, and Symphytocarpus

confluens.

Keyworps- Quercus bark, moist chamber technique

Introduction

The Marmara region of Turkey is located in the Euro-Siberian and the

Mediterranean phytogeographical regions of Turkey. The western part of the

region is subjected to Mediterranean climate and is covered with pine forests

and shrubs, whereas the eastern and northern parts have a continental climate

and are rich in oak forests (Atalay 2002).

Turkey is an important world distribution area for Quercus diversity (Gtinal

1997). There are 18 species of oak trees in Turkey (Segmen et al. 1995), of which

11 occur in the Marmara region.

The first myxomycetes for Turkey were reported by Harkénen & Uotila

(1983) and Harkonen (1987). Although the number of myxomycete species

worldwide is about 1000 (Lado 2001), the reported numbers for Turkey

are relatively low, around 232 (Sesli & Denchev 2014). With this paper we

contribute further knowledge on the myxomycetes in the country by presenting

eight species collected on bark of Quercus trees in the Marmara region, all new

records for Turkey.

Materials & methods

Field excursions were carried out in a randomised manner during June and October

of 2005-2008. All material was collected from the bark of Quercus species in different

182 ... Oran & Ergiil

provinces of Turkey’s Marmara region. Myxomycete fructifications were obtained from

moist chamber cultures in the laboratory as described in Stephenson & Stempen (1994).

The cultures were moistened with distilled water and kept under diffuse sunlight within

a temperature range of 22-25°C.

The cultures were examined under a dissecting microscope every day for a 4-week

period. Whenever developing fructifications were found, the bark pieces on which they

were found were removed from the Petri dish, allowed to dry slowly, and then placed

in herbarium boxes. The same chambers were then rewetted for another 4-week period

and examined as before. The specimens are stored in the authors’ personal collections

in the Department of Biology, Uludag University, Bursa, Turkey. Species have been

identified according to Castillo et al. (1997), Farr (1976, 1981), Ing (1999), Keller &

Brooks (1977), Lado & Pando (1997), Martin & Alexopoulos (1969), Mitchell (2013),

Nannenga-Bremekamp (1991), Stephenson & Stempen (1994), and Thind (1997).

Nomenclature follows Lado (2001).

Species recorded

Amaurochaete tubulina (Alb. & Schwein) T. Macbr. PLATE 1

Sporocarps aethaliate. Aethalia pulvinate, somewhat depressed. Sporothecae

covered by a thin, translucent cortex. Hypothallus thin, shining, prominent.

Capillitium branched, irregularly sprouting out from the base, branching and

anastomosing freely. Spore mass black. Spores pale olivaceous, 12-15 um

diam., finely warted.

SPECIMEN EXAMINED - ‘TURKEY, BILECIK/MERKEZ; Karadere-Sitliik road,

40°05’35.3”N 29°49’51.9”E, alt. 977 m, on bark of Quercus frainetto in oak forest,

22.7.2007, Oran 303-1.

COMMENTS—Amaurochaete is a typical genus in the order Stemonitales

characterized by a dendroid capillitium. Previously only a single species,

A. atra (Alb. & Schwein.) Rostaf., has been reported from Turkey (Sesli &

Denchev 2014). An examination of published records of A. tubulina indicates

that this species is frequently isolated from conifer bark, whereas our specimen

was recovered from Quercus bark in a moist chamber culture. Our specimen

appears to represent the first aethaliate structure of an Amaurochaete obtained

from a moist chamber culture.

{

e °

@ vr

PiaTE 1. Amaurochaete tubulina. I- aethalium; II- a. capillitium, b. spores; III- spores.

Corticolous myxomycetes new for Turkey ... 183

Amaurochaete tubulina somewhat resembles A. atra and A. comata G. Lister

& Brandza. It differs from A. atra, which is clearly distinguished by an early

fugacious aethelial wall, in its capillitial structure that widens in the axils and

is connected to the columella but not connected to neighbouring sporangia.

Amaurochaete tubulina is distinguished from A. comata by its sparsely

anastomosing free ends, spiny capillitia, and spores that are less strongly warted

on one side.

PLATE 2. Comatricha lenpipita:| I- sporocarps; I- a. are columella; III- a. columella; b.

capillitium; c. spinose capillitial ends; [V- a. columella, b. capillitium, c. spore; d. capillitial ends;

V- a. capillitium, b. reticulate spores.

Comatricha longipila Nann.-Bremek. PLATE 2

Sporocarps in small groups or solitary, 1.5-2.0 mm tall, nearly erect.

Hypothallus discoid or continuous under the group, brown and inconspicuous.

Stalk up to 33% of the total height, with a net of fibres at the base, often opaque

above. Sporotheca cylindrical or obovoid, 0.3-0.6 mm diam., brown. Peridium

usually completely fugacious but sometimes persistent as a collar around the

stalk apex. Columella merging into the capillitium just below the apex of the

sporotheca. Capillitium primary branches perpendicular to the columella,

hardly anastamosing, dichotomously branched with many, rather straight,

long, free ends at the periphery which are sometimes slightly swollen. Spores

pale red-brown, 6.0-7.0 um diam., finely warted.

SPECIMEN EXAMINED — TURKEY, KIRKLARELI/MERKEZ; Uskiip—Beypinari road 2 km,

41°45’05.7”N 27°26'32.4’E, alt. 403 m, on bark of a solitary Quercus pubescens tree in

agricultural land, 15.6.2006, Oran 91-2.

184 ... Oran & Ergiil

COMMENTS—Our specimen, as was the case for the type specimen, appeared

in a moist chamber culture prepared with a bark sample from Quercus. The

peridium occasionally remains as a permanent collar around the stalk. As

noted by Nannenga-Bremekamp (1991), “C. longipila is characterized by the

lax internal capillitium, with long free ends at the periphery and its small

spores.” However, Ing (1999) indicated that its small spores, lack of an outer

capillitial network, and long free ends separate Comatrica longipila from small

specimens of C. laxa Rostaf.

(3) ®e ®

oy 9°

7) » io)

@ @ e

..° (s)

PLATE 3. Diderma cinereum. I- individual sporocarps;

I- a. capillitium, b. peridium, c. spores; III- verruculose spores.

Diderma cinereum Morgan PLATE3

Sporocarps sessile, gregarious, subglobose, somewhat depressed, 0.3-0.5

mm diam., pearl-grey. Hypothallus inconspicuous. Peridium single, thin,

smooth, crustose. Capillitium of slender, dark threads, sparsely branched,

easily detached from the columella. Spore-mass black. Spores dark violet-grey,

9-11 um diam., verruculose.

SPECIMEN EXAMINED — TURKEY, BALIKESIR/MERKEZ; Konakpinar-Sépk6y road 5

km, 39°25’00.8”N 27°50’06.5”E, alt. 331 m, on bark of Quercus cerris, 28.7.2006, Oran

242-2.

ComMMENTS— There are various reports of the presence of a single or double

peridium in this species, but our specimen clearly has a single peridium. Ing

(1999) observed that D. cinereum is similar to D. spumarioides (Fr.) Fr. but

differs in having a single peridium, deciduous capillitium, and dark grey spores.

Diderma umbilicatum Pers. PLATE 4

Sporocarps short-stalked or sessile, pearl-grey. Peridium double, the outer

layer calcareous or occasionally limeless, adhering closely to the membranous

inner layer. Dehiscence irregular, leaving a ragged cup which shows white on

the inside. Columella prominent, pale, subglobose, smooth or rugose. Stalk

stout, reddish-brown, covered with white lime. Spores dark brown in mass,

purple brown in transmitted light, densely verruculose, 10-12 um diam.

SPECIMENS EXAMINED — TURKEY, CANAKKALE/YENICE; around Gonen dam,

39°57’41.1”N 27°25'52.2’E, alt. 191 m, on bark of Quercus cerris in composite forest,

Corticolous myxomycetes new for Turkey... 185

PiateE 4. Diderma umbilicatum. I- individual sporocarp;

II- a. peridial fragments, b. capillitium, c. spores; II- a. capillitium, b. spores.

29.7.2006, Oran 253-1; SAKARYA/GEYVE; Bozcagiz-Koru road, 4 km to Koru village,

40°3133”N 30°26’49’E, alt. 350 m, on bark of Quercus virgiliana in oak—-beech forest,

7.10.2007, Oran 266-1.

ComMENtTS—Diderma umbilicatum is included in Diderma subg. Leangium

(Link) Macbr. and has often been considered a variety of D. radiatum

(L.) Morgan (Nannenga-Bremekamp 1991). Ing (1999) suggested that

D. umbilicatum resembles both D. montanum (Meyl.) Meyl. and D. radiatum,

especially with respect to peridial characteristics. Hence, it was considered to

be synonymous with D. radiatum until quite recently. However, it is actually

quite different, having a number of features (e.g., irregular dehiscence, non-

cartilaginous peridium, pale columella, stout stalk) that are not present in

D. radiatum. Under a hand lens, it may resemble D. montanum, from which

it can be distinguished by its thicker and reddish short stalk, large columella,

and usually larger spores. Ing (1999) noted that its large pale sporangia,

conspicuous pale columella, and thick stalk easily separate D. umbilicatum

from other Diderma taxa.

Didymium lenticulare K.S. Thind & T.N. Lakh. PLATE 5

Sporocarps stipitate, up to 2 mm tall, scattered to loosely gregarious.

Hypothallus dark brown, rotate. Stalk 1-1.8 mm long, erect, grooved, tapered,

dark brown below, lighter and translucent above, scantily charged with lime

crystals, particularly at the base. Sporotheca erect or nodding, strongly

discoid, deeply umbilicate at the base, white, 0.5-0.8 mm diam. Peridium

membranous, hyaline, densely covered with lime deposits which form very

irregular ridges and plates. Columella absent. Capillitium of stout, subhyaline

threads, sparingly branched and anastomosed and with bead-like thickenings.

Spore-mass black. Spores deep violaceous brown by transmitted light, globose,

distinctly verrucose, also marked by conspicuous clusters of darker and bigger

warts, 9-11 um diam.

SPECIMEN EXAMINED — TURKEY, KIRKLARELI/PINARHISAR; Cayirdere—Akoren road

2 km, 41°42’19.3’N 27°31'15.7’E, alt. 269 m, on bark of Quercus frainetto in an oak

stand, 25.7.2006, Oran 179-2.

186 ... Oran & Ergiil

PiateE 5. Didymium lenticulare. I- individual sporocarp;

II- a. stipe, b. pseudocolumella, c. capillitial threads; d. spores;

III- a. crusted lime, b. capillitium, c. spores.

ComMMENTS—Didymium lenticulare very closely resembles D. clavus (Alb. &

Schwein.) Rabenh. but differs in a number of features, including its hyaline,

crustose peridium with thick ridges, longer stipe, more dispersed lime crystals

(especially in the lower region), lack of a columella, and spores that are larger,

lighter in color, and distinctly verrucose (Thind 1977).

This species also resembles D. squamulosum (Alb. & Schwein.) Fr. in its

possession of a hyaline peridium and similar spores but differs in its densely

thick calcareous deposits on the peridium, strongly discoid sporotheca, and

lack of a columella. In addition, its stipe is much longer and darker than that

typically observed in D. squamulosum. ‘The stipe of D. lenticulare is non-

calcareous, although it is sprinkled with lime crystals.

The stipe also differs from that of D. iridis (Ditmar) Fr., which has strongly

discoid sporangia that are deeply umbilicate below. The extraordinarily small

lime crystals of D. lenticulare also serve to distinguish it from allied species

(Thind 1977).

eS 3 ¢

‘ge fe. 6

PLatE 6. Physarum serpula. I- plasmodiocarp;

II- a. lime node, b. spores, c. peridium; III- a. lime node, b. spores.

Physarum serpula Morgan PLATE 6

Plasmodiocarpous, forming lines, rings, or a simple network, 0.2-0.4

mm wide, sometimes fusing laterally to form a broad fruiting surface, often

interspersed with globose sporocarps, dull yellow or ochraceous, sessile.

Hypothallus diffuse. Peridium single, thin, of closely encrusted lime globules,

Corticolous myxomycetes new for Turkey ... 187

without calcareous scales, fragile, membranous, persistent. Capillitium dense,

calcareous, the nodes numerous, large, angular, branching, pale yellow or

whitish, connected by short, hyaline threads or broad limy strands and then

almost badhamioid. Spore-mass dull black. Spores dark brown, verruculose,

with a paler and smoother area on one side, 10-13 um diam.

SPECIMEN EXAMINED - TURKEY, Bursa/NILUFER; around Alaaddinbey village,

40°12’10.7”N 28°53’45.2”E, alt. 78 m, on bark of a solitary Quercus robur tree in

agricultural land, 16.9.2007, Oran 312-2.

CoMMENTS—Physarum serpula was treated by Ing (1999) as part of a trio with

P. auriscalpium Cooke and P. decipiens M.A. Curtis, and Martin & Alexopoulos

(1969) observed that these three morphologically similar species were often

confused with one another. Physarum serpula is frequently confused with

P. auriscalpium, which differs by having a peridium dotted with prominent

glossy lime scales rather than the peridium evenly impregnated with lime in

P. serpula. Ing (1999) observed that P decipiens, which is more sporangiate than

P. auriscalpium or P. serpula, has a more badhamioid capillitium.

PLATE 7. Stemonitis inconspicua. 1) sporocarps; II- individual sporocarp; II- apical end of

sporotheca; IV- basal end of sporotheca; V-— inner net of capillitium; VI- surface net.

Stemonitis inconspicua Nann.-Bremek. PLATE 7

Sporocarps clustered on a common hypothallus, sometimes gregarious, total

height 2-8 mm. Hypothallus thin, silvery, red-brown by light microscope. Stalk

approximately 0.5 mm long, thin, black, opaque, shining. Sporothecae brown,

blunt cylindrical, 1.5-2.5 mm tall. Peridium fugacious, except sometimes at

the base, where red-brown fragments show a structure similar to that of the

spores. Columella tapered, dissipating just below the apex. Capillitium forming

approximately 3 meshes on the radius, consisting of slender threads variously

188 ... Oran & Ergiil

widened at the junctions, becoming extremely fine towards the periphery, the

surface net slender, irregular, fragmentary, with spiny free ends, dissipating

towards the apex. Spore mass dark brown. Spores pale red-brown, prominently

banded-reticulate with 2-3 meshes across the hemisphere, 8-9 um diam.

excluding the bands, which form a border approximately 0.5 um high in optical

section.

SPECIMENS EXAMINED — TURKEY, BILEcIK/MERKEZ; Karadere-Sitliik road, 3 km

to Siitltik, 40°05’35.3”N 29°49’51.9’E, alt. 977 m, on bark of Quercus frainetto in an

oak stand, 22.7.2007, Oran 3-1; CANAKKALE/MERKEZ; Can-Canakkale road, Kocalar

turnout, 40°02’03.7”N 26°46’57.1’E, alt. 469 m, on bark of Quercus frainetto in an oak

stand, 6.7.2005, Oran 303-4.

COMMENTS—Stemonitis inconspicua can be distinguished from the similar

S. foliicola Ing by its small size and banded-reticulate spores (Ing 1999).

os a

ae Ow

PLATE 8. Symphytocarpus confluens. I- pseudoaethalium; II- silver coloured hypothallus;

II- a. hypothallus and stipe, b. capillitium; IV- a. enlargements on capillitium, b. spores; V) spores.

Symphytocarpus confluens (Cooke & Ellis) Ing & Nann.-Bremek. PLATE 8

Pseudoaethalium 2-3 mm tall and 0.5-4 cm diam., deep black. Sporothecae

approximately 0.5 mm diam. Hypothallus silvery. Peridium fugacious except

for a number of more or less rounded plates, smooth and connected to the

capillitium. Columella absent or, if present, opaque, dissipating below the apex.

Capillitium a wide-meshed reticulum of thick, dark purple threads with small

axillary membranes, looped at the periphery. Spores dark purple-brown in

transmitted light, (10-)11-13 um diam., distinctly warted.

Corticolous myxomycetes new for Turkey ... 189

SPECIMEN EXAMINED — TURKEY, KIRKLARELI/MERKEZ; Derek6y-Kula road, around

Gegitagzi village, 41°57’34.9"N 27°21'43.4”E, alt. 624 m, on bark of Quercus petraea,

16.6.2006, Oran 110-1.

CoMMENTS—Symphytocarpus confluens, which is close to S. impexus Ing &

Nann.-Bremek, differs in its shorter, more elongated, deep black sporangia,

which occasionally possess an opaque columella that dissipates below the

apex of the sporangium (although it is not uncommon for the columella to be

absent). Hence, the sporangia of S. confluens are much deeper black in color

and have much larger spores that are at least 10 (usually =12) um in diameter

(Nannenga-Bremekamp 1991).

Discussion

Bark from Quercus trees sampled from 11 cities and six provinces in the

Marmara region were the source of our new records for Turkey: Balikesir

province (Diderma cinereum), Bilecik province (Amaurochaete tubulina,

S. inconspicua), Canakkale province (D. umbilicatum, S. inconspicua),

Bursa province, (P. serpula), Kirklareli province (C. longipila, D. lenticulare,

S. confluens), and Sakarya province (D. umbilicatum). As the distribution

shows, two species (Didymium umbilicatum, Stemonitis inconspicua) were

recovered from two different provinces.

The character of the tree bark structure is an important factor for myxomycete

distribution and productivity. Harkonen et al. (2004) concluded that a smooth

basic bark has a low water holding capacity and does not support a high

diversity of myxomycetes, bryophytes, and lichens. McHugh (1998) noted that

individual Quercus species and Quercus forests are the most productive sources

for myxomycetes. Bark from oak trees offers a suitable structure for sheltering

myxomycete spores. Stephenson (1989) noted that the considerable variation in

bark structure offers good potential microhabitats for corticolous myxomycetes.

High water holding capacities may benefit corticolous myxomycetes directly as

their active plasmodia are highly dependent on the availability of liquid water.

Acknowledgements

We thank Prof. S.L. Stephenson (University of Arkansas) and D.W. Mitchell for their

invaluable suggestions to our manuscript. We offer also sincere thanks for the effort and

meticulous attention to the nomenclature provided Dr. S.R. Pennycook. We thank also

AJE for article editing process.

Literature cited

Atalay I. 2002. Ecoregions of Turkey. Meta Basimevi, Izmir, Turkey.

Castillo A, Ilana C, Moreno G. 1997. A critical study of some Stemonitales. Mycological Research

101(11): 1329-1340. http://dx.doi.org/10.1017/S0953756297004103

Farr ML. 1976. Flora Neotropica, Monograph No:16. N.Y. Bot. Garden.

Farr ML. 1981. How to know the true slime molds. Wm. C. Brown Company Publishers. lowa USA.

190 ... Oran & Ergiil

Giinal N. 1997. Tiirkiyede Baslica Agag Tirlerinin Cografi Yayilislari, Ekolojik ve Floristik

Ozellikleri. Cantay Kitabevi, Istanbul, Turkey.

Harkonen M. 1987. Some additions to the knowledge of Turkish myxomycetes. Karstenia 27: 1-7.

Harkonen M, Uotila P. 1983. Turkish myxomycetes developed in moist chamber cultures. Karstenia

de Tord be

Harkonen M, Rikkinen J, Ukkola T, Enroth J, Virtanen V, Jaaskalainen K, Rinne E, Hiltunen L,

Piippo S, He X. 2004. Corticolous myxomycetes and other cryptogams on seven native tree

species in Hunan Province, China. Systematics and Geography of Plants 74: 189-198.

Ing B. 1999. The myxomycetes of Britain and Ireland. The Richmond Publishing Co. Ltd. England.

Keller HW, Brooks, TE. 1977. Corticolous myxomycetes VII: contribution toward a monograph of

Licea, five new species. Mycologia 69: 667-684. http://dx.doi.org/10.2307/3758857

Lado C. 2001. Nomenmyx: a nomenclatural taxabase of myxomycetes. Cuadernos de Trabajo de

Flora Micoldgica Ibérica 16.

Lado C, Pando E. 1997. Flora Mycologica Iberica. Vol. 2. CSIC, Madrid, Spain.

Martin GW, Alexopoulos CJ. 1969. The myxomycetes. Univ. of Iowa Press, Iowa.

McHugh R. 1998. Corticolous myxomycetes from Glen Mhuire, Co. Wicklow. Mycologist 12:

166-168. http://dx.doi.org/10.1016/S0269-915X(98)80072-4

Mitchell DW. 2013. Inventory of all-world myxomycetes, dichotomous and synoptic keys, etc.

Privately published by the author. DVD [updated].

Nannenga-Bremekamp NE. 1991. A guide to temperate myxomycetes. Biopress Limited. Bristol.

Segmen O, Gemici Y, Gérk G, Bekat L, Leblebici E. 1995. Tohumlu Bitkiler Sistematigi. Ege

Universitesi Fen Fakiiltesi Kitaplar Serisi. No. 116. Izmir, Turkey

Sesli E, Denchev CM. 2014. Checklists of the myxomycetes, larger ascomycetes, and larger

basidiomycetes in Turkey. 6th edn. Mycotaxon Checklists Online

(http://www.mycotaxon.com/resources/checklists/sesli-v106-checklist.pdf): 1-136.

Stephenson SL. 1989. Distribution and ecology of myxomycetes in temperate forest. II. Patterns

of occurrence on bark surface of living trees, leaf litter, and dung. Mycologia 81: 608-621.

http://dx.doi.org/10.2307/3760136

Stephenson SL, Stempen, H. 1994. Myxomycetes: a handbook of slime molds. Timber Press,

Portland, Oregon USA.

Thind KS. 1977. The myxomycetes of India. ICAR, New Delhi.

MY COTAXON

http://dx.doi.org/10.5248/130.191

Volume 130, pp. 191-196 January-March 2015

A new species of Lophodermium with

variously branched paraphyses

Hal-LIn Gu’, YA-FEI Xu, DAN-DAN Lv’,

SHI-JUAN WANG’, & YING-REN LIN?*

' School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

AxsstrRact — Lophodermium heterocladum, a new fungus developing on leaves of

Rhododendron cavaleriei, was collected from the Mount Sanqingshan National Park of Jiangxi

Province, China. Description, illustration, and comments are provided for this taxon. The

type collection is deposited in the Reference Collection of Forest Fungi of Anhui Agricultural

University, China (AAUF).

Key worps — foliicolous fungus, taxonomy, fungal diversity, Ericaceae

Introduction

During research on leaf-inhabiting fungi on members of the Ericaceae

in the Mount Sanqingshan National Park of China, one specimen of the

ascomycete family Rhytismataceae was obtained. Examination of that material

and comparison with previously described species (Lin et al. 2001a,b) led to

the conclusion that the specimen represents a new species of Lophodermium

Chevall. Fifty-seven Lophodermium species have previously been reported

from China (Hou et al. 2009, Luo et al. 2010, Shi et al. 2010, Chen et al. 2012,

Lin et al. 2012, Wang et al. 2012, Chen et al. 2013, Gao et al. 2013, Wang et al.

2014, Lu et al. 2015).

Materials & methods

The conidiomata and ascomata were examined macroscopically with a dissecting

microscope at 10—50x magnification. After rehydration in water for ca 10 min, fruiting

bodies were cut into 10-15 um thick sections using a freezing microtome. The sections

were mounted in water, KOH solution, Melzer’s reagent, cotton blue in water, or

lactophenol-cotton blue and examined microscopically. The colors of various structures

192 ... Gu &al.

were observed in water. Gelatinous sheaths surrounding ascospores and paraphyses

were examined in 0.1% (w/v) lactophenol-cotton blue. Measurements were made using

material in 5% KOH and from more than 30 paraphyses, asci, ascospores, conidiogenous

cells, and conidia for each specimen. Point and line integrated illustrations of external

status and internal structures of fruiting bodies were drawn using a microscopic drawing

device. The type collection is deposited in the Reference Collection of Forest Fungi of

Anhui Agricultural University, Hefei, China (AAUF).

Taxonomy

Lophodermium heterocladum H.L. Gu & Y.R. Lin, sp. nov. Figs 1-8

MycoBank MB 808959

Differs from Lophodermium implicatum by its variably shaped paraphyses that branch

1-2(-3) times, the presence of periphysoids and hyphal bridges, and a well-developed

subhymenium forming a strongly concave bowl shape.

TYPE: China, Jiangxi, Mount Sangingshan National Park, Huaiyushan, alt. ca 600 m, on

fallen leaves of Rhododendron cavaleriei H. Lév. (Ericaceae), 18 August 2012, Y.R. Lin, F.

Zhou & L. Zhang 2604 (Holotype, AAUF 68712).

Erymo oey: heterocladum, referring to variably branched paraphyses of the fungus.

ZONE LINES absent.

CoONIDIOMATA developing on upper side of leaves scattered in pale sandy

brown, subcircular or irregular, pale areas with an obvious edge. In surface

view, conidiomata 100-170 um diam., rounded or subrounded, black-

brown in the centre and the perimeter line of the conidioma, grey-brown

elsewhere, slightly raising the substratum surface, discharging spores through

an inconspicuous central ostiole. In median vertical section, conidiomata

intraepidermal, lenticular. UPPER WALL extremely poorly developed, composed

of a small amount of tiny angular cells. BASAL WALL dark brown, consisting of

1-3 layers of 3-5 um diam., thick-walled angular cells. SUBCONIDIOGENOUS

LAYER 4-10 um thick, comprised of colorless, thin-walled angular cells 1-3

um diam. TRICHOGYNES 32-38 x 2.5-3.2 um, cylindrical, tapering towards the

rounded apex, 1-3-septate in the lower half. ConrpIoPHORES not observed.

CONIDIOGENOUS CELLS 10-14 x 1.8-2.4 um, ampullaceous, colorless, usually

proliferating sympodially and percurrently. Conrip1A 4-6 x 0.8-1 um, hyaline,

aseptate, cylindrical, sometimes slightly curved, obtuse or round at the ends,

smooth-walled.

Ascomarta in similar positions to conidiomata on the host, scattered to clustered,

occasionally coalescent. In surface view, ascomata 350-680 x 170-360 um,

elliptical, ends round or obtuse, black, slightly shiny, with a clearly marked

outline, moderately raising the substratum surface, but slightly sunken

near the split, opening by a single longitudinal split which extends almost

to the ascomatal edge. Lips absent. In median vertical section, ascomata

intraepidermal. COVERING STROMA 12-20 um thick, mainly composed of

Lophodermium heterocladum sp. nov. (China) ... 193

Fics 1-7. Lophodermium heterocladum (holotype) on Rhododendron cavaleriei. 1. Habit on a leaf.

2. Detail of fruiting bodies. 3. Ascoma in median vertical section. 4. Portion of ascoma in median

vertical section. 5. Paraphyses, asci and ascospores. 6. Conidioma in vertical section. 7. Conidia.

194 ... Gu &al.

Fic. 8. Lophodermium heterocladum (holotype): variously branched paraphyses.

Lophodermium heterocladum sp. nov. (China) ... 195

textura angularis with dark brown to black, thick-walled cells 3-5 um diam.,

connecting to the basal stroma. There is a 6-8 um thick mulch which comprises

strongly carbonized tissue with no obvious cellular structure outside the top

of the covering stroma. Periphysoids lining the inner face of the covering

stroma, 5-14 x 1-1.5 um, cylindrical, not swelling at the apex, hyaline, straight

or slightly curved, 0-4-septate. BASAL STROMA dark-brown, consisting of 1-3

layers of thick-walled angular cells 3-7 um diam. The triangular space between

the covering stroma and basal stroma is filled with hyaline, thin-walled angular

cells 4-9 um diam. SUBHYMENIUM well developed, 15-22 um thick, strongly

concave and forming a bowl shape, composed of colorless textura angularis

and intricata. PARAPHYSES 130-155 x 1.5-2.2 um, filiform, hyaline, thinly

septate, mostly gradually or abruptly swollen to 4.2—-5.8 um at the apex,

branching 1-2(-3) times often near the top and in the middle or occasionally

in the positions not far from the base, sometimes with hyphal bridges near the

base between adjacent paraphyses, surrounded by a ca 0.5 um thick gelatinous

matrix. AScI ripening sequentially, 110-140 x 5.5-7.2 um, cylindrical, apex

round, short-stalked, thin-walled, J-, 8-spored. AscosporEs arranged in a

fascicle, 85-120 x 1.2-1.6 um, filiform, hyaline, aseptate, straight or slightly

curved, apex round or obtuse, hardly tapered towards the round or acute base,

with a thin gelatinous sheath.

HOST SPECIES, HABITAT, & DISTRIBUTION: Producing conidiomata and

ascomata on fallen leaves of Rhododendron cavaleriei. Known only from the

type locality, Jiangxi Province, China.

ComMENTs — Lophodermium heterocladum is very similar to L. implicatum

Y.R. Lin & Z.S. Xu on Rhododendron maculiferum subsp. anhweiense (E.H.

Wilson) D.F. Chamb. in the way ascomata and conidiomata are embedded and

in ascal shape and size. However, L. implicatum has ascomata on both surfaces

of the leaf, dark brown zone lines, the corner between the covering stroma

and basal stroma consisting of a very small amount of colorless, thin-walled

angular cells, paraphyses branched near the apex and entangled to form a light

yellow-brown epithecium 8-12 um thick, subhymenium composed of colorless

textura porrecta, and lacks periphysoids and hyphal bridges (Lin et al. 2001a).

Lophodermium rufum Y.R. Lin & K. Lion Rhododendron maculiferum subsp.

anhweiense is easily distinguished from the new species by its much larger

(620-1040 x 430-580 um) subcuticular ascomata, well-developed rufous lips,

covering and basal stroma consisting of textura angularis-epidermoidea, wider

(8—10.5 um) asci, shorter and wider (54—86 x 1.5—2 um) ascospores containing

many granules, unbranched paraphyses slightly gradually swollen at the apex

(Lin et al. 2001b).

196 ... Gu & al.

Acknowledgments

The authors are grateful to Dr J.E. Taylor and Dr M. Ye for pre-submission reviews

leading to the improvement of our manuscript, and to Mr FE Zhou and Ms L. Zhang

for the field investigations. This study was supported by the National Natural Science

Foundation of China (No. 31270065, 31170019).

Literature cited

Chen JL, Lin YR, Hou CL, Wang SJ. 2012 [“2011”]. Species of Rhytismataceae on Camellia spp.

from the Chinese mainland. Mycotaxon 118: 219-230. http://dx.doi.org/10.5248/118.219

Chen L, D.W. Minter, Wang SJ, Lin YR. 2013. Two new species of Rhytismataceae on fagaceous

trees from Anhui, China. Mycotaxon 126: 109-120. http://dx.doi.org/10.5248/126.109

Gao XM, Lin YR, Huang HY, Hou CL. 2013. A new species of Lophodermium associated with the

needle cast of Cathay silver fir. Mycological Progress 12: 141-149.

http://dx.doi.org/10.1007/s11557-012-0817-y

Hou CL, Piepenbring M. 2009. Lophodermium puerense sp. nov. on needles of Pinus yunnanensis vat.

tenuifolia from southwest Yunnan. Mycotaxon 107: 259-262. http://dx.doi.org/10.5248/107.259

Lin YR, Liu HY, Hou CL, Wang SJ, Ye M, Huang CL, Xiang Y, Yu SM. 2012. Flora fungorum

sinicorum, vol. 40, Rhytismatales [in Chinese]. Science Press, Beijing. 261p.

Lin YR, Xu ZS, Li K. 2001a. Two new species of the genus Lophodermium from the Huangshan

Mountains [in Chinese]. Mycosystema 20: 457-460.

Lin YR, Xu ZS, Li K, Xie YS. 2001b. Two new species of Lophodermium Chev. on Anhwei

rhododendron [in Chinese]. Journal of Anhui Agricultural University 28: 358-361.

Lu DD, Tang YP, Wang LH, Wang SJ, Lin YR. 2015 [“2014”]. Lophodermium quadrisporum sp.

nov. (Rhytismataceae) on Rhododendron faberi subsp. prattii. Mycotaxon 129(2): 459-463.

http://dx.doi.org/10.5248/129.459

Luo JT, Lin YR, Shi GK, Hou CL. 2010. Lophodermium on needles of conifers from Yunnan Province,

China. Mycological Progress 9:235-244. http://dx.doi.org/10.1007/s11557-009-0632-2

Shi GK, Luo JT, Hou CL. 2010. Species of the Rhytismataceae on Pinus armandii [in Chinese].

Mycosystema 29(2): 159-163.

Wang SJ, D.W. Minter, Gao XM, Lin YR. 2012. A new species of Lophodermium on Saccharum

arundinaceum [in Chinese]. Mycosystema 31(4): 471-475.

Wang SJ, Xu YF, Tang YP, Li Q, Lin YR. 2014. Lophodermium urniforme, a new species of Rhytismataceae

from China. Mycosystema 33(4): 768-772. http://dx.doi.org/10.13346/j.mycosystema. 140029

MY COTAXON

http://dx.doi.org/10.5248/130.197

Volume 130, pp. 197-205 January-March 2015

Bertia hainanensis sp. nov. (Coronophorales)

from southern China

LARISSA N. VASILYEVA ', HAI-XIA MA?,

ALEKSEY V. CHERNYSHEV>", & STEVEN L. STEPHENSON?

‘Institute of Biology & Soil Science, Far East Branch of the Russian Academy of Sciences,

Vladivostok 690022, Russia

?Institute of Tropical Bioscience & Biotechnology, Chinese Academy of Tropical Agricultural Sciences,

Haikou 571101, China

°A.V. Zhirmunsky Institute of Marine Biology, Far East Branch of the Russian Academy of Sciences,

Vladivostok 690059, Russia

‘Far Eastern Federal University, Vladivostok 6900950, Russia

*Department of Biological Sciences, University of Arkansas, Fayetteville, Arkansas 72701, USA

* CORRESPONDENCE TO: vasilyeva@biosoil.ru

ABSTRACT — A new species of Bertia collected in southern China is described and illustrated.

Bertia hainanensis is characterized by a combination of such features as non-ostiolate

ascomata with a roughened tuberculate surface, 1-septate cylindrical-geniculate ascospores,

and filiform paraphyses. It is most similar to B. tropicalis, from which it differs by its narrower

ascospores and its filiform paraphyses. The distinctive features of the ascomatal surface were

examined with a scanning electron microscope. The validity of the family Bertiaceae is

discussed.

KEY worpDs — Ascomycota, coronophoralean fungi, SEM images, taxonomy

Introduction

In 2012, the second author collected a specimen of Bertia De Not. from

the Bawangling National Nature Reserve (Hainan Province, China), and we

consider it to represent a species new to science. As in our previous studies

on several other coronophoralean fungi (Vasilyeva et al. 2010, 2012a, 2013),

we prepared images of fruiting bodies using a scanning electron microscope

(SEM).

There are only a few records of Bertia species from China. Bertia macrospora

Sacc. and B. moriformis (Tode) De Not. have been reported from Gansu, Jiangsu,

198 ... Vasilyeva & al.

Jilin, and Yunnan provinces (Teng 1996); B. sinensis J.C. Krug & Corlett was

described from Yunnan (Krug & Corlett 1988) and B. biseptata Sivan. & W.H.

Hsieh from Taiwan (Hsieh et al. 1995).

Materials & methods

The type specimen of the new species is deposited in the Fungarium of the Chinese

Academy of Tropical Agricultural Sciences, Haikou, Hainan, China (FCATAS). Material

was prepared for SEM examination in accordance with Vasilyeva et al. (2012a). Light

micrographs of asci and ascospores were obtained using a Leica DM 4500B microscope,

and a Zeiss EVO 40 scanning electron microscope was used to examine surface details

of the ascomata.

Taxonomy

Bertia hainanensis Lar.N. Vassiljeva, HX. Ma, Chernyshev &

S.L. Stephenson, sp. nov. Fie. 1

MycoBank MB 857516

Differs from Bertia tropicalis by its narrower ascospores and its filiform paraphyses.

Type: China, Hainan Province, Bawangling National Nature Reserve, on dead branches

of an unidentified tree, 5 November 2012, Hai-Xia Ma (Holotype: FCATAS 300).

EryMo_oey: refers to the province of China where the fungus was collected.

AsScOMATA superficial, mostly aggregated, turbinate, with a thick sterile base,

collapsing collabent when dry, black, 640-820 um diam., with a roughened,

tuberculate surface, tubercules 50-75 x 35-50 um.; walls 40-60 um thick,

consisting of 3-4 layers of angular cells 7-11 x 6-8 um; Munk pores present,

0.8-1 um diam. Asci elongated-clavate, 8-spored, long-stipitate, with a kind

of apical ring ca. 2 um, in the spore-bearing portions (72-)80-100 x 14-18

um, stalks 100-120 um, oozing from ascomata in fascicles intermingled with

numerous filiform paraphyses. Ascosporgs overlapping biseriate, cylindrical,

basal one-third curved, geniculate, 1-septate, hyaline, 24-28 x 4.8-5.5 um.

Notes: The most similar species described in the literature appears to be Bertia

tropicalis Huhndorf et al., which differs by its wider ascospores (5-9.5 um) and

the presence of inflated paraphyses (Huhndorf et al. 2004). Bertia tropicalis

is known from Costa Rica, French Guiana, Jamaica, Panama, and the United

States (Puerto Rico), all localities that are consistent with a Caribbean center

of fungal biodiversity. As such, B. tropicalis could be considered as a vicarious

species with respect to B. hainanensis, which might be expected to occur in

other countries within the southeastern Asian center of fungal diversity (cf.

Vasilyeva et al. 2012b). Bertia hainanensis differs from both B. tropicalis and

B. sinensis in its more narrow ascospores but shares with B. tropicalis the

collapsing collabent ascomata in contrast to the elongated and non-collapsing

ascomata that characterize B. sinensis.

Bertia hainanensis sp. nov. (China) ... 199

Fic.1. Bertia hainanensis (holotype, FCATAS 300): a-d. Ascomata; e. Ascomatal surface; f, g. Cross

section through a lateral wall; h. Fascicle of asci and paraphyses; i. Ascus with ascospores; j. Ascal

apex showing a tiny apical ring; k. Ascospores; |. Paraphyses. Scale bars: a = 0.5 mm; b, c = 200 um;

d, h = 100 um; e, i= 50 um; f, k, 1 = 20 um; g, j =5 um.

Other members of the relatively small group of Bertia species known from

southeastern Asia are B. fructicola Henn. and B. oxyspora (Penz. & Sacc.) Hohn.

from Indonesia (Hennings 1894a; Penzig & Saccardo 1897), B. novoguineensis

200 ... Vasilyeva & al.

Henn. from the northeast of the island of New Guinea (Hennings 1894b), and

B. tessellata Petch and B. turbinata Petch from Sri Lanka (Petch 1922). Almost

all of these have wider ascospores than B. hainanensis, with B. oxyspora also

characterized by longer ascospores (<50 um). Only B. tessellata has an ascospore

width (4.5-5.0 um) that is similar to that of B. hainanensis, but ascospores of

B. tessellata are pale fuscous and 3-septate.

Discussion: some remarks on the Bertiaceae

Both the genus Bertia and the family Bertiaceae Smyk are poorly investigated,

and their current composition is uncertain. Smyk (1981) proposed the family

without referring to its distinguishing characters, although she cited the

cytological studies of ascomata and asci by several French mycologists (Luc

1952, Chadefaud 1954, Parguey-Leduc 1977) as warranting segregation of Bertia

into a new family. As circumscribed by Smyk (1981), the new family included

three genera (Bertia, Calyculosphaeria Fitzp., and Rostrocoronophora Munk),

but Calyculosphaeria species are now considered to belong to Nitschkia G.H.

Otth ex P. Karst., while Rostrocoronophora has been transferred to a different

order (Diaporthales) as a synonym of Gnomonia Ces. & De Not. (Bolay 1972).

One might wonder just which characters could support the family Bertiaceae

sensu Smyk.

A search for characters in the French literature cited by Smyk (1981) also

has not revealed the possible criteria for distinguishing the family Bertiaceae.

All three publications compared B. moriformis with species from other—

sometimes very distantly related—groups, and such a comparison has surely

provided a number of differences, but if a new family is segregated within the

order Coronophorales, as in the case of the study by Smyk (1981), it should be

based upon differences among the members of this order. Luc (1952) compared

B. moriformis and Systremma natans (‘Tode) Theiss. & Syd. [= Dothidea sambuci

(Pers.) Fr., Dothideales| and listed several characters between them (e.g., asci

with double walls thickened at the apex versus asci with single and thin walls

without a thickening at the apex), but these very same characters distinguish the

roughly outlined groups Ascohymeniales and Ascoloculares (Nannfeldt 1932).

So, although those characters properly separate B. moriformis and S. natans

into higher taxonomic groups, they cannot serve as a basis for delimiting the

family Bertiaceae since different hierarchical levels of a taxonomic system are

characterized by their own characters. In addition, Luc (1952) compared the

asci of B. moriformis with those found in Chaetomium Kuntze and Melanospora

Corda (although without referring to particular species) and concluded that B.

moriformis might be an ‘intermediate’ taxon that occupies a position between

the order Dothideales and the class Haerangiomycetes as created (Falck & Falck

1947) for Melanospora and Ophiostoma Syd. & P. Syd. However, this clearly

Bertia hainanensis sp. nov. (China) ... 201

represents an example of unfairly judging the “phylogenetic” position of

B. moriformis without properly analyzing numerous groups, and such a peculiar

comparison cannot be considered as the basis for creating the Bertiaceae.

Chadefaud (1954) compared B. moriformis with another dothidealean

species, Dothidella ulmi (C.-J. Duval) G. Winter. His conclusions were similar

to those by Luc (1952), and B. moriformis was suggested as being separate from

the Dothideales, but one cannot estimate the level of differences with only two

species under comparison. Parguey-Leduc (1977) discussed different modes

of ascus formation along with the structure of ascus wall and apical ring in

pyrenomycetous fungi. She mentioned B. moriformis only once, indicating

that its dangeardium (ie., the ascus initial) is two-celled (a rare condition

for the pyrenomycetes). However, the same dangeardium was also observed

in Cordyceps militaris (L.) Fr., Epichloe typhina (Pers.) Tul & C. Tul., and

Mycosphaerella bolleana B.B. Higgins, so that feature might also represent a case

of parallelism in different fungal groups — a relatively frequent phenomenon.

Again, one cannot find any reason to warrant the Bertiaceae as a separate family.

Quite recently Huhndorf et al. (2004) accepted the Bertiaceae because two

species (B. moriformis and B. tropicalis) form a well-supported clade on the

molecular tree of the Coronophorales. However, two circumstances should be

taken into consideration. First, a taxon cannot exist without delineating the

differences that distinguish it from other taxa at the same hierarchical level.

Second, the level of ‘clades’ (familial, subfamilial, etc.) is unknown without an

estimation of differences between tentatively segregated groups. As has been

pointed out, taxa occupying the same taxonomic level are characterized by a

state combination of the same characters (Vasilyeva & Stephenson 2012, 2013).

Still more recently, Mugambi & Huhndorf (2010) provided the necessary

distinguishing characters for the Bertiaceae, but once again these appear to

be unsatisfactory. Members of this family are supposed to have ascomata that

are mostly tuberculate (if smooth, then the ascospores are uniformly brown)

and ascospores longer than 15 um. However, ascospore size cannot be used

for discriminating among families, since this character is universally used

for species delimitation. As for the presence of tubercules, these structures

characterize many members of the Coronophorales and differ only in size and

shape (Vasilyeva et al., 2010, 2012a, 2013). Moreover, the absence of tubercules

is also permitted in the Bertiaceae, so this character clearly cannot be a criterion

for the differentiation of the family.

In general, the simple accumulation of differences for the circumscription of

a group is often useless, since it is not the number of characters but the weight

of each character that actually matters. Very often, a single difference might be

of great value. As pointed out elsewhere (Vasilyeva & Stephenson 2012), the

weight of a character is its position in the taxonomic hierarchy, and this weight

202 ... Vasilyeva & al.

depends upon the frequency at which a character is used to delimit groups at

a certain level. Judging from the characters that have been proposed for the

segregation of the Bertiaceae, we have no choice but to abandon this family

until more substantial characters are found. The position of Bertia within

Nitschkiaceae Nannf. seems acceptable for the time being, although there is a

question about the differences between Bertia and Nitschkia.

For example, Nitschkia macrospora Teng has rather large and rough tubercules

on the stromatal surface (Vasilyeva et al. 2010), a condition almost comparable

with that found in B. latispora (Corlett & J.C. Krug) Lar.N. Vassiljeva (Fic. 2).

Moreover, N. macrospora ascospores are large (30-45 um long), so this species

appears to fit the concept of Bertia, at least sensu Mugambi & Huhndorf (2010).

On the other hand, B. orbis Mugambi & Huhndorf (Mugambi & Huhndorf

2010: Fig. 5r) has ascospores that are typical for Nitschkia. The clustering of

B. orbis with other Bertia species apart from Nitschkia species in the molecular

tree (Mugambi & Huhndorf 2010: Fig. 1) is not sufficient evidence supporting

their independence. In general, the dichotomous structure of molecular trees

strongly influences the composition of clades.

To illustrate this situation, we can use one of the ascomycetous families. If one

keeps the combinatorial variation of fungi in mind and attempts to construct

a dichotomous tree of the genera of the family Gnomoniaceae, the type genus

Gnomonia will fall in the same clade as Apiognomonia, Gnomoniella, and

Ophiognomonia when ascomatal type is used to define this clade. In contrast,

Gnomonia will appear in the same clade as Chalcosphaeria, Melanopelta, and

Plagiostoma when ascospore type found in each genus is considered (Vasilyeva

& Stephenson 2010). Molecular data might support both arrangements (if

they are correlated with defining features) or point towards new ones, but it

is highly probable that all genera in the Gnomoniaceae comprise a network of

taxa instead of reflecting a cladistic structure, with several characters that are

of equal value defining genera in the combinatorial space. The same situation

might be observed among the genera that comprise the Coronophorales, and we

have already pointed out the partial combinatorial variation within this order

(Vasilyeva et al. 2013). The same conclusion could be reached by considering

the key to genera provided by Mugambi & Huhndorf (2010), where several

differences (e.g., asci 8-spored vs. multispored and ascomata spinulose vs. not

spinulose) are repetitive in the delimitation of genera. The choice of any such

equal-leveled characters will lead to clades with different compositions in the

dichotomous ‘phylogenetic’ trees.

An independent existence of Bertia might be warranted in part by the

presence of a peculiar kind of ascospore—they are wormlike or cylindrical and

curved geniculate, similar to those found in some species of Lasiosphaeria Ces.

& De Not., including such examples as L. hirsuta (Fr.) Ces. & De Not., L. hispida

Bertia hainanensis sp. nov. (China) ... 203

d. Ascospores. Scale bars: a = 200 um; b = 100 um; c = 50 um; d = 20 um.

(Tode) Fuckel, and L. ovina (Pers.) Ces. & De Not. Ascospore types have been

traditionally used to segregate genera in ascomycetous fungi, and we can follow

this tradition for the time being.

Acknowledgments

We are grateful to the engineer D.V. Fomin for his assistance in providing the

micrographs produced at the Far Eastern Center of Electron Microscopy of the A.V.

Zhirmunsky Institute of Marine Biology (Vladivostok). We also thank Prof. Makoto

Kakishima (Jilin Agricultural University, China) and Dr. Adam Rollins (Lincoln

204 ... Vasilyeva & al.

Memorial University, USA) for serving as presubmission reviewers and for providing

helpful comments and suggestions.

Literature cited

Bolay A. 1972 [“1971”]. Contribution 4 la connaissance de Gnomonia comari Karsten. Etude

taxonomique, phytopathologique et recherches sur sa croissance in vitro. Berichte der

Schweizerischen Botanischen Gesellschaft 81: 398-482.

Chadefaud M. 1954. Sur les asques de deux Dothideales. Bulletin de la Société Mycologique de

France 70: 99-108.

Falck R, Falck O. 1947. A new class of Ascomycetales. A contribution to the orbis vitae system of

fungi. Palestine Journal of Botany, Rehovot series, 6: 89-106.

Hennings P. 1894a. Neue und interessante Pilze aus dem K6nigl. botanischen Museum in Berlin

II. Hedwigia 33: 229-233.

Hennings P. 1894b. Fungi novo-guineenses II. Botanische Jahrbiicher fiir Systematik,

Pflanzengeschichte und Pflanzengeographie 18, Beiblatt 44: 22-40.

Hsieh WH, Chen CY, Sivanesan A. 1995. Taiwan fungi: new species and new records of ascomycetes.

Mycological Research 99: 917-931. http://dx.doi.org/10.1016/S0953-7562(09)80750-8

Huhndorf SM, Miller AN, Fernandez FA. 2004. Molecular systematics of the Coronophorales

and new species of Bertia, Lasiobertia and Nitschkia. Mycological Research 108: 1384-1398.

http://dx.doi.org/10.1017/S0953756204001273

Krug JC, Corlett M. 1988. A new species of Bertia from China. Canadian Journal of Botany 66:

1256-1258. http://dx.doi.org/10.1139/b88-179

Luc M. 1952. Structure et développement de deux Dothideales: Systremma natans (Tode) Th. et Syd.

et Bertia moriformis (Tode) de Not. Bulletin de la Société Mycologique de France 68: 149-164.

Mugambi GK, Huhndorf SM. 2010. Multigene phylogeny of the Coronophorales: morphology and

new species in the order. Mycologia 102: 185-210. http://dx.doi.org/10.3852/09-043

Nannfeldt JA. 1932. Studien uber die Morphologie und Systematik der nicht-lichenisierten

inoperculaten Discomyceten. Nova Acta Regiae Societatis Scientarum Upsaliensis, ser. 1V 8(2):

1-368.

Parguey-Leduc A. 1977. Les asques de Pyrénomycetes. Revue de Mycologie 41: 281-339.

Penzig O, Saccardo PA. 1897. Diagnoses fungorum novorum in insula Java collectorum. Malpighia

11: 387-409.

Petch T. 1922. Additions to Ceylon fungi (II). Annals of the Royal Botanic Gardens, Peradeniya

Ziad 9-322.

Smyk LV. 1981. Fungi of the order Coronophorales in the mycoflora of the Ukraine. Ukrainian

Journal of Botany 38(6): 46-49 [in Ukrainian].

Teng SC. 1996. Fungi of China. Mycotaxon, Ltd., Ithaca. 586 p.

Vasilyeva LN, Stephenson SL. 2010. The problems of traditional and phylogenetic taxonomy of

fungi. Mycosphere 1: 45-51.

Vasilyeva LN, Stephenson SL. 2012. The hierarchy and combinatorial space of characters in

evolutionary systematics. Botanica Pacifica 1: 21-30.

Vasilyeva LN, Stephenson SL. 2013. An essentialistic view of the species problem. 141-169, in: IY

Pavlinov (ed.). The species problem. Ongoing issues. Rijeka: InTech.

Vasilyeva LN, Chernyshev A, Stephenson SL. 2010. Pyrenomycetes of the Russian Far

East 4: family Nitschkiaceae (Coronophorales, Ascomycota). Mycologia 102: 233-247.

http://dx.doi.org/10.3852/09-090

Bertia hainanensis sp. nov. (China) ... 205

Vasilyeva LN, Chernyshev AV, Stephenson SL. 2012a. Neochaetosphaerella thaxteriospora gen. et

sp. nov. and Tympanopsis texensis sp. nov. (Coronophorales, Ascomycota) from Texas, USA.

Fungal Diversity 52: 191-196. http://dx.doi.org/10.1007/s13225-011-0124-y

Vasilyeva LN, Stephenson SL, Hyde KD, Bakhali AH. 2012b. Some stromatic pyrenomycetous

fungi from northern Thailand - 1. Biscogniauxia, Camillea and Hypoxylon (Xylariaceae).

Fungal Diversity 52: 65-76. http://dx.doi.org/10.1007/s13225-011-0150-9

Vasilyeva LN, Chernyshev AV, Stevenson SL, Hyde KD. 2013. Tortulomyces thailandicus gen. et sp.

nov. and Nitschkia siamensis sp. nov. (Coronophorales, Ascomycota) from northern Thailand.

Mycoscience 54: 110-115. http://dx.doi.org/10.1016/j.myc.2012.09.004

MYCOTAXON

http://dx.doi.org/10.5248/130.207

Volume 130, pp. 207-213 January-March 2015

Distribution of Alternaria species among sections. 1.

Section Porri

PHILIPP B. GANNIBAL

Laboratory of Mycology and Phytopathology, All-Russian Institute of Plant Protection,

Shosse Podbelskogo 3, Saint Petersburg, 196608, Russia

CORRESPONDENCE TO: phbgannibal@yandex.ru

ABSTRACT — The evaluation of all available descriptions of Alternaria species allowed the

assignment of another 71 species to A. sect. Porri, in addition to the 46 species previously

included on the basis of phylogenetic analysis. We assess that this section encompasses 117

species, which are listed. An emended and somewhat expanded description of A. sect. Porri

is presented.

Key worps — Nimbya, N. rhapontici

Introduction

The genus Alternaria Nees is a large taxonomic group that comprises

approximately 280 species (Simmons 2007). A series of large-scale works

has attempted to resolve the phylogeny of Alternaria and other alternarioid

hyphomycetes by employing more than ten different genomic loci (Pryor &

Bigelow 2003; Hong et al. 2005; Runa et al. 2009; Lawrence et al. 2012, 2013,

2014; Woudenberg et al. 2013). Several well-supported phylogenetic lineages

have been revealed within the alternarioid hyphomycetes that have led to

several taxonomic novelties. The genus Alternaria was divided into eight

taxonomic sections by Lawrence et al. (2013). According to an alternative

concept (Woudenberg et al. 2013) all alternarioid hyphomycetes were collapsed

into one genus, Alternaria, with 24 sections, 11 of which referred to Alternaria

sensu stricto.

The monophyletic status of the so-called large-spored species was supported

by all analyses. Both research groups (Lawrence et al. 2013; Woudenberg et

al. 2013) agreed that the group of large-spored species should be treated as a

well-defined section of the genus, Alternaria sect. Porri, based on phylogenetic

as well as morphological data. Forty-six species were included in the section

208 ... Gannibal

by Lawrence et al. (2013). However, the size of the section was estimated to

comprise c. 120 (Gannibal 2012) or c. 80 (Woudenberg et al. 2013) species.

Alternaria porri was chosen as type species. Only one large-spored species,

A. helianthiinficiens E.G. Simmons et al., was placed apart from section Porri

(Woudenberg et al. 2013).

The main discriminating morphological features of many Alternaria sect.

Porri species are larger conidia and the presence of a long filiform apical beak

(Lawrence et al. 2013, Woudenberg et al. 2013). Sexual morphs have not yet

been discovered. Probably all species in this section are parasites of plants.

Species of A. sect. Porri have a rather strong substrate specialization and usually

infect either a single plant species or only few species within a single family.

It is unlikely that all 280 Alternaria species will be molecularly characterized

in the foreseeable future. Moreover, for several species, living isolates are not

known and herbarium material is mostly too old and scanty for successful

DNA extractions. On the other hand, the morphological diversity of Alternaria

is well studied and available in a monograph published by Simmons (2007).

Classical comparative morphology can be utilized to establish the affiliation of

all Alternaria species to a specific section.

The aim of this work was to detect among phylogenetically unexamined

Alternaria species, as far as possible, all species that fit the current morphological

circumscription of A. sect. Porri and to emend the morphological concept of

this section to reflect its extended membership.

Materials & methods

Morphology of almost all Alternaria species with legitimate names was analyzed

with regard to conformity with criteria of Alternaria sect. Porri. The morphological

assessment was based predominantly on descriptions made by Simmons (2007).

Species described after 2007 were assessed using original diagnoses and illustrations.

Descriptions of eight species included in A. sect. Porri were found in Sun & Zhang 2007

(A. lactucicola and A. rhaponticicola); Labuda et al. 2008 (A. jesenskae); Zhang & Zhang

2008 (A. amphicarpaeae, A. sojae, and A. tribuli); and Gannibal 2010 (A. simmonsii

and A. silybi). Two conceptions of A. sect. Porri were used for comparison, those of

Lawrence et al. (2013) and Woudenberg et al. (2013).

Results & discussion

The evaluation of all available descriptions of Alternaria species allowed for

the addition of 71 species to A. sect. Porri. The size of this section is assessed to

comprise 117 species. Alternaria species of A. sect. Porri are listed in TABLE 1.

Eight species for which living specimens are not available—A. bannaensis

W.Q. Chen & TY. Zhang, A. basellae T.Y. Zhang, A. calystegiae Nelen, A. ellisii

Pandotra & Ganguly, A. iridicola (Ellis & Everh.) J.A. Elliott, A. nicotiana J.L.

Cheng, A. nyctanthis K.B. Deshp. & Rajd., and A. trachelospermicola (T.Y. Zhang

Alternaria sect. Porri...

TABLE 1. List of Alternaria spp. assigned to A. sect. Porri.

> PPB BP BS

BPPeeeeereaereeererreeeeeePer eee e eer eszee eee eee eee

acalyphae (Nelen) E.G. Simmons [a]

. acalyphicola E.G. Simmons (Lawrence et al. 2013)

. africana E.G. Simmons [b]

. agerati Sawada ex E.G. Simmons (Lawrence et al. 2013)

. agripestis E.G. Simmons & K. Mort. (Lawrence et al. 2013)

_ allii Nolla

. amphicarpaeae Meng Zhang & T.Y. Zhang

. anagallidis A. Raabe (Lawrence et al. 2013)

. anodae E.G. Simmons

. aragakii E.G. Simmons (Lawrence et al. 2013)

. argyroxiphii E.G. Simmons & Aragaki (Lawrence et al. 2013)

. ascaloniae E.G. Simmons & C.F. Hill

. azadirachtae E.G. Simmons & Alcorn

. bataticola Ikata ex W. Yamam. (de Hoog & Horre 2002; Lawrence et al. 2013)

. beringelae E.G. Simmons

. beticola E.G. Simmons & C.F. Hill

. blumeae E.G. Simmons & Sontirat (Hong et al. 2005; Lawrence et al. 2013)

. bonducellae R. Dubey et al. [b]

. brasiliensis .M. Queiroz et al.

. calendulae Ondiej (Lawrence et al. 2013)

. capsici E.G. Simmons (Hong et al. 2005; Lawrence et al. 2013)

. caricae T.Y. Zhang et al. [b]

. carthami S. Chowdhury (Lawrence et al. 2013)

. cassiae Jurair & A. Khan (Lawrence et al. 2013)

. catharanthi (T.Y. Zhang & X.E. Lin) E.G. Simmons [b]

. centaureae E.G. Simmons

. cepulicola V.G. Rao [b]

. ciceris E.G. Simmons

. cichorii Nattrass (Lawrence et al. 2013)

. cirsinoxia E.G. Simmons & K. Mort. (Lawrence et al. 2013)

. crassa (Sacc.) Rands (Pryor & Bigelow 2003; Hong et al. 2005; Lawrence et al. 2013)

. cretica E.G. Simmons & Vakal. (Lawrence et al. 2013)

. cucumericola E.G. Simmons & C.F. Hill

cucumerina (Ellis & Everh.) J.A. Elliott (Hong et al. 2005; Lawrence et al. 2013)

. cyamopsidis Rangaswami & A.V. Rao

. cyphomandrae E.G. Simmons (Lawrence et al. 2013)

. danida E.G. Simmons (Lawrence et al. 2013)

. daturicola T.Y. Zhang et al. [b]

. dauci (J.G. Kiihn) J.W. Groves & Skolko (Pryor & Bigelow 2003; Hong et al. 2005;

Lawrence et al. 2013; Woudenberg et al. 2013)

. dichondrae Gambogi et al. (Lawrence et al. 2013)

. echinaceae E.G. Simmons & C.F. Hill

. eichhorniae Nag Raj & Ponnappa

. enydrae S.A. Khan & M. Kamal [b]

. euphorbiae (Barthol.) Aragaki & J.Y. Uchida [b]

. euphorbiicola E.G. Simmons & Engelhard (Hong et al. 2005; Lawrence et al. 2013)

A. flagelloidea (G.F. Atk.) Luttr. [b]

A. gaurae E.G. Simmons & C.F. Hill

209

210 ... Gannibal

A. glyceriae E.G. Simmons & C.F. Hill

A. grandis E.G. Simmons (Lawrence et al. 2013)

A. guangxiensis W.Q. Chen & T.Y. Zhang [b]

. hawaiiensis E.G. Simmons (Lawrence et al. 2013)

. heliophytonis E.G. Simmons

. herbiculinae E.G. Simmons

. hibiscinficiens E.G. Simmons & C.F. Hill

. hordeiseminis E.G. Simmons & G.F. Laundon

. hortensiae Unamuno ex E.G. Simmons [b]

. iranica E.G. Simmons & Ghosta

. jesenskae Labuda et al.

. lactucicola X. Sun & T.Y. Zhang

. lallemantiae (Khokhr.) E.G. Simmons [c]

. latispora T.Y. Zhang & Meng Zhang [b]

. limicola E.G. Simmons & M.E. Palm (Lawrence et al. 2013)

. linariae (Neerg.) E.G. Simmons

. linicola J.W. Groves & Skolko (de Hoog & Horre 2002; Lawrence et al. 2013)

. longirostrata T.Y. Zhang & Meng Zhang [b]

. loofahae E.G. Simmons & Aragaki

. lunariae (Oudem. & C.J.J. Hall) E.G. Simmons [b]

. macrospora Zimm. (Pryor & Bigelow 2003; Hong et al. 2005; Lawrence et al. 2013;

Woudenberg et al. 2013)

. montanica E.G. Simmons & Robeson

. multirostrata E.G. Simmons & C.R. Jacks. (Lawrence et al. 2013)

. nattrassii E.G. Simmons [b]

. nitrimali E.G. Simmons & M.E. Palm (Lawrence et al. 2013)

. novae-guineensis E.G. Simmons & C.F. Hill

. obtecta E.G. Simmons

A. passiflorae J.H. Simmonds (Lawrence et al. 2013)

A. petasitis M. Kubota et al. [b]

A. pipionipisi E.G. Simmons

A. poonensis Ragunath (Lawrence et al. 2013)

A. porri (Ellis) Cif. (Pryor & Bigelow 2003; Hong et al. 2005; Lawrence et al. 2013;

Woudenberg et al. 2013)

A. protenta E.G. Simmons (Lawrence et al. 2013)

A. pseudorostrata E.G. Simmons (Hong et al. 2005; Lawrence et al. 2013; Woudenberg et

al. 2013)

A. pulcherrimae T.Y. Zhang & J.C. David

. ranunculi E.G. Simmons

. readeri (G. Winter) Neerg. [b]

. rhaponticicola X. Sun & T.Y. Zhang

. ricini (Yoshii) Hansf. (Lawrence et al. 2013)

. rosa-sinensis M.X. Gao & T.Y. Zhang [b]

. rosifolii E.G. Simmons & C.F. Hill

. rostellata E.G. Simmons (Lawrence et al. 2013)

. saposhnikoviae J.Z. Zhang & T.Y. Zhang [b]

. sauropodis E.G. Simmons

. scorzonerae (Aderh.) Loer. (Lawrence et al. 2013)

. sesami (E. Kawam.) Mohanty & Behera (de Hoog & Horre 2002; Lawrence et al. 2013)

SPP PP PPR PPR RBBB eB

> P PPB Se BBP BB

Alternaria sect. Porri... 211

. sidae E.G. Simmons

. silybi Gannibal

. simmonsii Gannibal

. sojae Meng Zhang & TY. Zhang

. solani Sorauer (Pryor & Bigelow 2003; Hong et al. 2005; Lawrence et al. 2013;

Woudenberg et al. 2013)

. solani-nigri R. Dubey et al. (Lawrence et al. 2013)

. spinaciae Allesch. & F. Noack [b]

. stachytarpheticola E.G. Simmons [b]

. steviae Ishiba et al. (Lawrence et al. 2013)

. subcylindrica E.G. Simmons & R.G. Roberts (Lawrence et al. 2013)

. subtropica E.G. Simmons [b]

. sudanensis E.G. Simmons [b]

. tabasco E.G. Simmons & R.G. Roberts

. tagetica S.K. Shome & Mustafee (Lawrence et al. 2013; Woudenberg et al. 2013)

. thunbergiae E.G. Simmons & Alcorn

. tillandsiae E.G. Simmons & C.F. Hill

. tomato (Cooke) L.R. Jones [d]

. tomatophila E.G. Simmons (Lawrence et al. 2013)

. tribuli Meng Zhang & T.Y. Zhang

. tropica E.G. Simmons (Lawrence et al. 2013)

. vanuatuensis E.G. Simmons & C.F. Hill

. venezuelensis E.G. Simmons & Rumbos

. viciae-fabae E.G. Simmons & G.F, Laundon

. zinniae M.B. Ellis (Lawrence et al. 2013)

> > SB DB

SPPPPP PRR PEEP PBB eeBeB

Bold font names = species with phylogenetic data (in the cited references).

Unannotated names = species with reliable living cultures and herbarium specimens.

Names with square bracketed annotations = species with herbarium specimens as the only

available reliable material:

[a] = representative isolates have not been chosen;.

[b] = no known living isolates;

[c] = isolates are non-sporulating;

[d] = isolates are not correctly identified.

et al.) E.G. Simmons—were found to have intermediate characteristics that do

not allow unambiguous placement in A. sect. Porri or any other section. Dubious

morphological traits and the absence of isolates for detailed morphological or

molecular studies make it impossible to assess their phylogenetic positions

within the genus.

An isolate of A. tomato (CBS 114.35, deposited in 1935) clustered with strong

support in the autonymous clade, Alternaria sect. Alternaria (Lawrence et al.

2013). However, the credible species description in Simmons (2007) suggests

that this species resides in A. sect. Porri. According to Simmons (2007), no

correctly identified A. tomato isolates are known. Hence, we suggest that

A. tomato sensu Simmons belongs in A. sect. Porri.

Phylogenetic data revealed the polyphyletic nature of the genus Nimbya E.G.

Simmons (Lawrence et al. 2012). Only four studied species from Cyperaceae

212 ... Gannibal

and Juncaceae clustered together as a separate clade (“true” Nimbya) when three

species from Amaranthaceae were placed in Alternaria sect. Alternantherae

(Lawrence et al. 2012, 2013). There are five Nimbya species found on plants

other than Cyperaceae, Juncaceae, and Amaranthaceae, which likely belong

to other clades. They have relatively large conidia and some of them could be

related to the group of large-spored Alternaria species. At least N. rhapontici

(Nelen) E.G. Simmons is morphologically very similar to Alternaria sect.

Porri, but additional examinations are necessary to clarify the phylogeny and

taxonomy of this species.

Here we present an emended and somewhat expanded description of the

section:

Alternaria sect. Porri D.P. Lawr., Gannibal, Peever & B.M. Pryor, Mycologia 105:

541. 2013; emend. Gannibal.

On V-8 primary conidiophores short, moderately long (up to 150-250 um)

or rarely very long (up to 700 um or probably even longer [A. pseudorostrata,

A. nitrimali]), simple or branched, with one or few (up to 9 on branched

conidiophore [A. solani]) conidiogenous loci. Conidia solitary, in short

or rarely moderately long chains (up to 7 units on PCA [A. pseudorostrata,

A. euphorbiicola]), simple or with solitary lateral conidia, body of mature

conidia moderately large (40-110 x 11-25(-30) um), broadly ovoid, obclavate,

ellipsoid, subcylindrical, or obovoid, disto- and euseptate, slightly constricted at

septa, number of transepta in the conidial body varying from 7-9 [A. cirsinoxia]

to 12-19 [A. crassa, A. grandis], with a single septum or 2-3 longitudinal

septa in one or almost all transverse divisions; conidia of some species almost

always without any longitudinal septa [A. agripestis, A. subcylindrica], pale

yellowish tan to dark olivaceous brown, smooth or punctulate, at least some

mature conidia with terminal beaks, solitary or in groups (up to 5) appearing as

several beaks or a single beak with several branches, beaks aseptate or septate,

50-300(-460) um long and (4—)5-6(-8) um wide near the base, apex tapering

to 1.5-3 um or giving rise in secondary conidiophores. Conidia may form

lateral or apical secondary conidiophores without filiform portion, sometimes

with one or only few lateral secondary conidiophores arising from the body or beak.

In comparison with V-8, on PCA conidia are somewhat larger and have

beaks with larger average numbers of branches (Simmons 2007; Gannibal et

al. 2014).

Acknowledgments

It is my pleasure to acknowledge the attention of Dr. Uwe Braun and Dr. Daniel

Lawrence for their presubmission reviews of this article. This work was supported by

Russian Science Foundation (project #14-26-00067).

Alternaria sect. Porri... 213

Literature cited

Gannibal PB. 2010. Taxonomic studies of Alternaria from Russia: new species on Asteraceae.

Mycotaxon 114: 109-114. http://dx.doi.org/10.5248/114.109

Gannibal PB. 2012. Chapter 11. Understanding the phylogeny of the alternarioid hyphomycetes:

what can the consequences be in taxonomy? 303-331, in: JK Misra et al. (eds). Systematics and

Evolution of Fungi. Science Publishers Inc., Enfield, New Hampshire, USA.

Gannibal PB, Orina AS, Mironenko NV, Levitin MM. 2014. Differentiation of the closely related

species, Alternaria solani and A. tomatophila, by molecular and morphological features and

aggressiveness. Eur. J. Plant Pathol. 139: 609-623. http://dx.doi.org/10.1007/s10658-014-0417-6

Hong SG, Cramer RA, Lawrence CB, Pryor BM. 2005. Alt a 1 allergen homologs from Alternaria

and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genet. Biol.

42: 119-129. http://dx.doi.org/10.1016/j.fgb.2004.10.009

de Hoog GS, Horré R. 2002. Molecular taxonomy of the Alternaria and Ulocladium species

from humans and their identification in the routine laboratory. Mycoses 45: 259-276.

http://dx.doi.org/10.1046/j.1439-0507.2002.00747.x

Labuda R, Elias P Jr, Sert H, Sterflinger K. 2008. Alternaria jesenskae sp. nov., a new species

from Slovakia on Fumana procumbens (Cistaceae). Microbiol. Res. 163: 208-214.

http://dx.doi.org/10.1016/j.micres.2006.05.004

Lawrence DP, Park MS, Pryor BM. 2012. Nimbya and Embellisia revisited, with nov.

comb. for Alternaria celosiae and A. perpunctulata. Mycol. Progress 11: 799-815.

http://dx.doi.org/10.1007/s11557-011-0793-7

Lawrence DP, Gannibal PB, Peever TL, Pryor BM. 2013. ‘The sections of Alternaria: formalizing

species-group concepts. Mycologia. 105(3): 530-546. http://dx.doi.org/10.3852/12-249

Lawrence DP, Gannibal PB, Dugan FM, Pryor BM. 2014. Characterization of Alternaria

isolates from the infectoria species-group and a new taxon from Arrhenatherum,

Pseudoalternaria arrhenatheria sp. nov. Mycol. Progress 13(2): 257-276.

http://dx.doi.org/10.1007/s11557-013-0910-x

Pryor BM, Bigelow DM. 2003. Molecular characterization of Embellisia and Nimbya species and

their relationship to Alternaria, Ulocladium and Stemphylium. Mycologia 95: 1141-1154.

http://dx.doi.org/10.2307/3761916

Runa F, Park MS, Pryor BM. 2009. Ulocladium systematics revisited: phylogeny and taxonomic

status. Mycol. Progress 8: 35-47. http://dx.doi.org/10.1007/s11557-008-0576-y

Simmons EG. 2007. Alternaria. An identification manual. Utrecht: CBS. 775 pp.

Sun X, Zhang T-Y. 2007. Taxonomic studies of Alternaria 10: Two new species and a new record

from China. Mycotaxon 101: 283-287.

Woudenberg JHC, Groenewald JZ, Binder M, Crous PW. 2013. Alternaria redefined. Studies in

Mycology. 75: 171-212. http://dx.doi.org/10.3114/sim0015

Zhang M, Zhang T-Y. 2008. Taxonomic studies of Alternaria from China 11. Three large-spored

new species. Mycotaxon. 103: 263-268.

MY COTAXON

http://dx.doi.org/10.5248/130.215

Volume 130, pp. 215-221 January-March 2015

Morchella galilaea, an autumn species from Turkey

HatTirA TASKIN™, Hasan HUseyIn DoGAn?, & SAADET BUYUKALACA'

‘Department of Horticulture, Faculty of Agriculture, University of Cukurova,

01330 Adana, Turkey

*Department of Biology, Faculty of Science, University of Selcuk

42074, Konya, Turkey

*CORRESPONDENCE TO: hatirataskin1@gmail.com

Asstract — Morchella specimens collected during the autumn seasons of 2009-12 from

Adana Province, Turkey, have been identified by DNA sequencing as representatives of

Morchella galilaea. The macro- and micromorphology, phylogeny, ecology, and fruiting

phenology of the Turkish material are evaluated.

KEY worps — mist units, morels, taxonomy, volcanic media

Introduction

Morchella species (true morels) are some of the most valuable and important

fungi consumed worldwide due to their unique aroma and flavor. Their

commercial value has been increasing over the years as a result of growing

consumer demand. Countries with diverse morel species include China, USA,

Pakistan, India, and Turkey, which export morels to European countries (Pilz

et al. 2007). In Turkey, morels generally grow in pine forests (Pinus brutia,

P. nigra) from late February to June (Taskin & Bityiikalaca 2012). According to

a review by Pilz et al. (2007), researchers have reported Morchella associated

with elm, ash, aspen, tulip poplar, apple, and cottonwood trees. Although morel

fruiting season occurs generally during the spring season, Sturgis (1905) and

Masaphy et al. (2009) have reported morels fruiting in different seasons (Pilz et

al. 2007). Morels may be found throughout Turkey, especially in Mediterranean

and Aegean regions. If they can be collected in relatively large volumes, morels

offer the potential of providing a significant income for people living near the

forests and could be regarded as a significant export product for Turkey.

In addition to the existence of an endemic Turkish morel species,

M. anatolica \siloglu et al. (Isiloglu et al. 2010), Taskin et al. (2010, 2012)

216 ... Taskin, Dogan, & Bityiikalaca

identified 20 phylogenetically distinct Morchella species in Turkey. Nineteen

of these species were collected from forests in spring, but one (given the

phylogenetic code Mes-16) was observed in autumn in the mist units of the

Horticultural Research and Application Area of Cukurova University, Adana,

Turkey (Taskin et al. 2012). Based on studies by Taskin et al. (2012), O'Donnell

et al. (2011), Du et al. (2012a,b) we named this population as a new species

but later realized that it represented Morchella galilaea, described from Israel

by Clowez (2012). However, as detailed morphological, microscopical, and

ecological features were not provided in the original description, we have

evaluated the morphology, phylogenetic position, ecology, and fruiting

phenology of M. galilaea collected from Turkey. Our results should be beneficial

to future studies concerned with culture and conservation of Morchella spp.

Materials & methods

Specimens were collected from Adana province in Turkey (Horticultural Research

and Application Area of Cukurova University) during autumn and early winter

(October-December) between 2009 and 2012 (Fic. 1). Two (in 2009), two (in 2010),

and 11 (in 2012) Morchella specimens were collected from mist units used for rooting

agricultural plant cuttings.

Molecular analyses were performed at the United States Department of Agriculture

(USDA-NCAUR-Peoria-IL) in USA (Taskin et al. 2012). These samples were analyzed

through partial RNA polymerase I (RPB1), RNA polymerase II (RPB2), translation

elongation factor (EFl-a), 28S (LSU) rRNA gene and internal transcribed spacer

(ITS) rRNA gene sequences. Macro- and microscopical characters were studied at

the Department of Biology, Selcuk University (Konya, Turkey). The specimens were

mounted in Melzer’s reagent and examined microscopically using a Leica DM 750

microscope. Volcanic tuff was analyzed at Alata Horticultural Research Institute, and

pH (Jackson 1962), salinity (Soil Survey Division Staff 1993), total lime (CaCO,; Caglar

1949), organic matter (Walkley & Black 1934), phosphorus (P; Olsen & Dean 1965),

magnesium (Mg), calcium (Ca), iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu)

were analyzed using an atomic absorption spectrophotometer (Varian SpectrAA Model

220FS, Germany).

Taxonomy

Morchella galilaea Masaphy & Clowez, Bull. Soc. Mycol. Fr. 126 (3-4): 238.

2012 [“2010”]. Fics 1-2

ASCOMATA 53-78 mm tall. HymMENopHORE 36-45 mm tall, 12.5-17.5

mm wide, conic to subcylindric, pitted and ridged, ridges have two types,

these are narrow polygonal ridges and long elliptical ridges. STERILE RiBs

longitudinally arranged, 5.2-9.8 mm wide, 14.5-24.74 mm tall, thin, elastic,

simple, or forked and anastomosing to form elongated hymenial pits, white,

bluntly rounded when young, but often flattened in age, ridges glabrous.

Morchella galilaea in Turkey ... 217

Fic. 1 - Morchella galilaea specimens collected from mist units

on the Cukurova University campus (Adana, Turkey).

HyMENIAL Pits light gray, whitish gray, blackish gray or silvery gray, when

young; becoming, greyish-light brown, light olive brown or yellow brown with

maturity. ASCOCARP MARGIN connecting directly into the stalk without a sharp

bend, free. Stipe 17-33 mm high, 5-8.2 mm wide, cylindrical and thickening

slightly towards the base; surface when young completely glabrous and white,

in maturity light yellowish white. CoNTExT whitish, 0.5-1 mm thick in the

hollow hymenophore; sterile inner surface whitish, glabrous. AscosPORES

elliptical, smooth (surface distinctly rugulate to tectate under the scanning

electron microscope), with homogeneous contents, (12.5-)13.5-18(-19.5)

x (7.5-)8-9.5(-10) um [average Q =1.83]. Asci 8-spored, 165-220 x 15-22

um, cylindric, hyaline. PARAPHYSES cylindric to subclavate, apices rounded to

subacute, with 2-3-septate, hyaline or with brown to brownish homogeneous

contents in KOH (2%), 100-130 x 6-11 um. RESIDUAL PARAPHYSES on sterile

ridges similar to paraphyses, hyaline or with bundled brownish contents.

SPECIMENS COLLECTED: Turkey. Adana: Cukurova University Campus, 37°01’55”N

35°22'12”E, alt. 39 m, solitary or scattered on volcanic tuff in misted cutting propagation

beds, 17 Nov 2009, leg. H. Taskin, Taskin 117 (ANK); 28 Nov 2012, leg. H.H. Dogan,

Dogan 10022 (Kon Fungarium).

218 ... Taskin, Dogan, & Bityiikalaca

EHT=20.00kKV ——Signal A= SE1 Mag= 6.00 KX

WD = 10.5 mm IProbe= 20pA

Fic. 2 - Morchella galilaea (Turkish specimen, Taskin 117).

A, B: asci with ascospores. C: paraphyses. D: ascospore (SEM).

Discussion

The morphological features of M. galilaea were cited by Clowez (2012) as:

ascomata 3-5 cm height, ovoid, light yellow; primary alveolus large, few; edges

of ribs same color and rusty; stipe cylindrical rather slender, attenuated at the

top and wider at the base.

When we compared M. galilaea samples collected from Israel and Turkey

morphologically, we found certain differences between the samples, which

we summarize as follows: (a) ascomata of the Israel specimens were shorter

(3-5 cm); (b) the stipes of the Turkish specimens were more or less cylindrical

except for the slightly thicker base while in the Israeli collection the stipes had

a narrower apex and wider base; (c) the Israeli specimen was found under

Fraxinus syriaca, while our Cukurova University specimens were collected

from a no tree zone.

Morchella galilaea in Turkey ... 219

Molecular characterization

Morchella galilaea [as Mes-16] has been molecularly assessed through

GCPSR analyses by Taskin et al. (2012) and also molecularly analyzed by

O’Donnell et al. (2011), Du et al. (2012 a, b), Masaphy et al. (2010), Clowez

(2012), and Richard et al. (2015). These studies indicate that M. galilaea occurs

in China (Du et al. 2012a,b), Java, Hawaii, Israel (Masaphy et al. 2010, Clowez

2012), New Zealand, India (Kanwal et al. 2011), and three African countries

as well as in Turkey. The Turkish and Israel sequences of Morchella galilaea

are deposited in GenBank-ITS1-5.8S-ITS2 as JQ723082 and GU589858,

respectively (Masaphy et al. 2010; Clowez 2012; Du et al. 2012 a, b).

Ecological assessment

The results of the volcanic tuff analyses are provided in TABLE 1. Relatively

high levels of copper, manganese, zinc, iron, magnesium and calcium were

detected in the volcanic tuff medium, which was identified as having a loam

soil structure, normal CaCO,, and alkali salinity profile. There was a relatively

low potassium level. Singh et al. (2004), who determined that pH values vary

from 6.5 to 7 in morel natural areas in India, detected high levels of calcium,

carbon, nitrogen, sodium, and lead and low levels of phosphates, chloride, and

potassium; soil type varied from sandy to loam.

Sturgis (1905) observed morels fruiting in September in an aspen-spruce

forest burned the previous summer in southwestern British Columbia (Pilz

et al. 2007). Masaphy et al. (2009) reported that a Morchella species emerged

from late autumn to late spring in northern Israel under Rhamnus alaternus,

TABLE 1. Properties of the volcanic tuff in which Morchella galilaea was found fruiting.

PROPERTY RESULTS

Texture (100 g/ml) 46

Caco, (%) 19.6

Salinity (mmhos/cm) 0.06

Organic matter (%) 2.6

pH 7.6

Available potassium (ppm) 167.5

Available phosphorus (ppm) 19.2

Calcium (ppm) 2300

Available magnesium (ppm) 759

Iron (ppm) 6.6

Zinc (ppm) 2.8

Manganese (ppm) 4.8

Copper (ppm) 1.6

220 ... Taskin, Dogan, & Bityiikalaca

Platanus orientalis, Populus euphratica, Laurus nobilis, and Fraxinus syriaca.

However, they could not find a mycorrhizal relationship between trees and

morels. Morchella is known to be saprobic. However, according to a review study

conducted by Tedersoo et al. (2010), Morchella species may associate weakly

with plant roots and thus are likely to be facultative biotrophs. Nonetheless,

some researchers believe that certain Morchella species may be able to form

mycorrhizal relationships with certain trees. Dahlstrom et al. (2000) reported

a mycorrhizal relationship between Morchella and species of Pinaceae, and

Buscot & Kottke (1990) detected a mycorrhizal association between Morchella

and Picea abies. Our morel specimens grew in a mist unit. While this unit was

used for rooting olive cuttings during 2009-2010, no cuttings were placed

into the units in 2012, which suggests that our specimens are probably not

mycorrhizal. Our hypothesis is that the mycelium of the species was carried

to the mist units by means of the volcanic tuff, where it developed over the

years. After mycelial development, a sprinkle irrigation system and hormone

application used for rooting might also have stimulated Morchella fruiting. The

same species has been found under Populus and Quercus by Du et al. (2012b)

and under Fraxinus syriaca by Masaphy et al. (2010). Although morels typically

fruit from late February to early June, our Morchella specimens were collected

between October and December. In 2012, the air temperature at Cukurova

University-Adana averaged 29°C in October, 23°C in November, and 16°C in

December.

Acknowledgements

The authors would like to express their sincere gratitude to Dr. Kerry O'Donnell

(USDA-NCAUR- Peoria, IL, USA) for allowing us to perform molecular analyses in his

laboratory. A special thanks goes to Dr. Davut Keles (Manager of Alata Horticultural

Research Institute), for the volcanic tuff analysis, and to Berken Cimen, for organization

of the figures. We are also indebted to Prof. Dr. Mitko Karadelev, Prof. Dr. Aysun Peksen,

Prof. Dr. Daniel Royse for their critical reviews of the manuscript.

Literature cited

Buscot F, Kottke I. 1990. The association of Morchella rotunda (Pers.) Boudier with roots of Picea

abies (L.) Karst. New Phytologist 116: 425-430.

Caglar KO. 1949. Soil science. Ankara University Agriculture Faculty Publication, Ankara.

Clowez P. 2012 (‘2010’). Les morilles: une nouvelle approche mondiale du genera Morchella.

Bulletin de la Société Mycologique de France 126: 199-376.

Dahlstrom JL, Smith JE, Weber NS. 2000. Mycorrhiza-like interaction by Morchella with species of

the Pinaceae in pure culture synthesis. Mycorrhiza 9: 279-285.

Du XH, Zhao Q, Yang ZL, Hansen K, Taskin H, Bitytikalaca S$, Dewberry D, Moncalvo JM, Douhan

GW, Robert V, Crous PW, Rehner SA, Rooney AP, O'Donnell K, Sink S. 2012a. How well do

ITS rDNA sequences ‘barcode’ species of true morels (Morchella)? Mycologia 104: 1351-1368.

http://dx.doi.org/10.3852/12-056

Morchella galilaea in Turkey... 221

Du XH, Zhao Q, Yang ZL, O'Donnell K, Rooney AP, Yang ZL. 2012b. Multigene molecular

phylogenetics reveals true morels (Morchella) are especially species-rich in China. Fungal

Genetics and Biology 49: 455-469. http://dx.doi.org/10.1016/j.fgb.2012.03.006

Isiloglu M, Alli H, Spooner BM, Solak MH. 2010. Morchella anatolica (Ascomycota), a new species

from southwestern Anatolia, Turkey. Mycologia 102: 455-458. http://dx.doi.org/10.3852/09-186

Jackson ML. 1962. Soil chemical analysis. Prentice Hall, New York.

Kanwal HK, Acharya K, Ramesh G, Reddy MS. 2011. Molecular characterization of Morchella

species from the western Himalayan region of India. Current Microbiology 62: 1245-1252.

http://dx.doi.org/10.1007/s00284-010-9849-1

Masaphy S, Zabari L, Goldberg D. 2009. New long season ecotype of Morchella rufobrunnea from

northern Israel. Micologia Aplicada International 21: 45-55.

Masaphy S, Zabari L, Goldberg D, Jander-Shagug G. 2010. The complexity of Morchella systematics:

a case of the yellow morel from Israel. Fungi 3(2): 14-18.

O’Donnell K, Rooney AJ, Mills GL, Kuo M, Weber NS, Rehner SA. 2011. Phylogeny and historical

biogeography of true morels (Morchella) reveals an early Cretaceous origin and high continental

endemism and provincialism in the Holarctic. Fungal Genetics and Biology 48: 252-265.

http://dx.doi.org/10.1016/j.fgb.2010.09.006

Olsen SR, Dean LA. 1965. Estimation of available phosphorus in soils by extraction with sodium

bicarbonate. United States Department of Agriculture, Washington, D.C.

Pilz D, Mclain R, Alexander S, Villarreal-Ruiz L, Berch S, Wurtz TL, Parks CG, McFarlane E, Baker

B, Molina R, Smith JE. 2007. Ecology and management of morels harvested from the forests of

western North America. General Technical Report, PNW-GTR-710. United States Department

of Agriculture, Forest Service, Pacific Northwest Research Station, Portland.

Richard F, Sauve M, Bellanger JM, Clowez P, Kansen H, O'Donnell K, Urban A, Courtecuisse

R, Moreau PA. 2015. True morels (Morchella, Pezizales) of Europe and North America:

Evolutionary relationships inferred from multilocus data and a unified taxonomy. Mycologia

http://dx.doi.org/10.3852/14-166.

Singh SK, Kamal S, Tiwari M, Rai RD, Upadhyay RC. 2004. Myco-ecological studies of natural morel

bearing sites in Shivalik Hills of Himachal Pradesh, India. Micologia Aplicada International 16:

1-6.

Soil Survey Division Staff. 1993. Soil survey manual. United States Department of Agriculture

Handbook 18. Soil Conservation Service.

Sturgis WC. 1905. Remarkable occurrence of Morchella esculenta (L.) Pers. Journal of Mycology

Like2.69;

Taskin H. Biyitikalaca S. 2012. Morel (Morchella) mushroom. Bah¢e 41: 25-36.

Taskin H, Buyiikalaca $, Dogan HH, Rehner SA, O’Donnell K. 2010. A multigene molecular

phylogenetic assessment of true morels (Morchella) in Turkey. Fungal Genetics and Biology 47:

672-682. http://dx.doi.org/1016/j.fgb.2010.05.004

Taskin H, Bityitikalaca S, Hansen K, O’Donnell K. 2012. Multilocus phylogenetic analysis of true

morels (Morchella) reveals high levels of endemics in Turkey relative to other regions of Europe.

Mycologia 104: 446-461. http://dx.doi.org/10.3852/11-180

Tedersoo L, May TW, Smith M. 2010. Ectomycorrhizal lifestyle in fungi: global diversity,

distribution, and evolution of phylogenetic lineages. Mycorrhiza 20: 217-263.

Walkley A, Black LA. 1934. An examination of the Degtjareff method for determining soil organic

matter and a proposed modification of the chromic acid titration method. Soil Science 39:

29-38.

MY COTAXON

http://dx.doi.org/10.5248/130.223

Volume 130, pp. 223-225 January-March 2015

Hyphoderma hallenbergii,

a new corticioid species from India

MANINDER Kaur, AVNEET P. SINGH’, & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT — Hyphoderma hallenbergii sp. nov. from Himachal Pradesh, India is described.

Key worps — Baghi, Basidiomycota, Agaricomycetes, Meruliaceae

During August 2012 fungal forays conducted of the Baghi area of district

Shimla, Himachal Pradesh, India, Maninder and Avneet collected a fungus

on decaying wood of Cedrus deodara in a mixed forest. Comparison of

macroscopic and microscopic characters (Eriksson & Ryvarden 1975, Dhingra

1989, Larsson 1998, Bernicchia & Gorjén 2010, Singh et al. 2010, Priyanka

& Dhingra 2012, Mycobank 2014) suggested that it belonged in the genus

Hyphoderma (Meruliaceae, Agaricomycetes, Basidiomycota). The fungus, which

is similar to H. nemorale K.H. Larss. and H. incrustatum K.H. Larss. in having

two types of cystidia but which differs in basidiospore size, is described here as

a new species.

Hyphoderma hallenbergii Man. Kaur, Avneet P. Singh & Dhingra, sp.nov. Plate 1

MycoBank MB 809801

Differs from Hyphoderma nemorale and H. incrustatum by its smaller and ellipsoid

basidiospores.

Type: India, Himachal Pradesh: Shimla, Narkanda, on Baghi road, on the bark of

decaying wood of Cedrus deodara (Roxb. ex D. Don) G. Don, 19 August 2012, Maninder

Kaur & Avneet 6962 (PUN, holotype).

Erymo.oey: In the honor of Dr. Nils Hallenberg, Professor Emeritus, University of

Gothenburg, Sweden.

Basidiocarp resupinate, adnate, effused, <100 um thick in section; hymenial

surface smooth to somewhat grandinioid, chalky white; margins thinning

224 ... Kaur, Singh, & Dhingra

9, aS yo > SIN

a8. g

PiatTE 1. Hyphoderma hallenbergii (holotype). 1. Basidiocarp showing hymenial surface.

2. Capitate hymenial cystidia. 3. Basidium. 4,5. Basidiospores.

6. Vertical section through basidiocarp.

out or indeterminate, concolorous or paler. Hyphal system monomitic.

Generative hyphae <2.5 um wide, branched, septate, clamped; subiculum

poorly developed, formed of few hyphae running more or less parallel to

the substrate; subhymenial hyphae well developed, vertical, denser. Cystidia

of two types: [1] Capitate cystidia 25-44 x 5-6 um, subfusiform, originating

from the hymenium, thin-walled, with basal clamp, sparsely encrusted; [2]

Cystidia which are cylindrical to flexuous with constrictions, sometimes

almost moniliform 60-80 x 5-7 um, originating from subhymenium, slightly

thick-walled, with basal clamp, often encrusted with crystals. Basidia 16-34 x

4-5 um, clavate to subclavate, constricted to sinuous, 4-sterigmate, with basal

clamp; sterigmata <3.5 um long. Basidiospores 6.3-7.5 x 3.5-4.5 um, ellipsoid,

with oily contents, thin-walled, inamyloid, acyanophilous.

REMARKS — Hyphoderma nemorale and H. incrustatum both differ from

H. hallenbergii by their larger, cylindrical basidiospores (H. nemorale, 10-14 x

4-5 um; H. incrustatum, 11-16 x 4-5.5 um; Larsson 1998).

Hyphoderma hallenbergii sp. nov. (India) ... 225

Acknowledgements

The authors thank Head (Department of Botany, Punjabi University, Patiala) for

providing research facilities, Dr. Nils Hallenberg (Professor Emeritus, Denmark)

for expert comments and peer review; and Prof. B.M. Sharma (Department of Plant

Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer review.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.1. Fungi Europaei 12. Edizioni Candusso. Alassio.

Italia. 1008 p.

Dhingra GS. 1989. Genus Hyphoderma Wallr. em. Donk in the Eastern Himalayas. 197-212, in: ML

Triveda et al. (eds). Plant Science Research in India. Today & Tomorrow’s Printers & Publishers,

New Delhi.

Eriksson J, Ryvarden L. 1975. The Corticiaceae of North Europe- II. Fungiflora, Oslo. pp. 288-546.

Larsson KH. 1998. Two new species in Hyphoderma. Nordic Journal of Botany. 18(1): 121-127.

http://dx.doi.org/10.1111/j.1756-1051.1998.tb01106.x

MycoBank. 2014. Fungal databases. Nomenclature and species banks. [Accessed: 16/07/2014]

http://www.mycobank.org/.

Priyanka, Dhingra GS. 2012. Two new species of Hyphoderma (Agaricomycetes) from India.

Mycotaxon 119: 255-260. http://dx.doi.org/10.5248/119.255

Singh AP, Priyanka, Dhingra GS, Singla N. 2010. A new species of Hyphoderma (Basidiomycetes)

from India. Mycotaxon 111: 71-74. http://dx.doi.org/10.5248/111.71

MY COTAXON

http://dx.doi.org/10.5248/130.227

Volume 130, pp. 227-231 January-March 2015

Xylaria thailandica — a new species from

southern Thailand

NATTHAPACH SRIHANANT™, VASUN PETCHARAT?’,

& LARISSA N. VASILYEVA?

‘Tropical Agricultural Resource Management Programme & Department of Pest Management,

Faculty of Natural Resources, Prince of Songkla University,

Hatyai, Songkhla, 90112, Thailand

*Institute of Biology and Soil Sciences, Far East Branch of the Russian Academy of Sciences,

Vladivostok 690022, Russia

* CORRESPONDENCE TO: srihanant.n@gmail.com

ABSTRACT —A new species, Xylaria thailandica, was collected from dead wood and fallen

branches in southern Thailand. This fungus has upright, gregarious, cylindrical stromata that

arise from a tomentose stipe and which is internally dark brown to black at the center but

yellow around the perithecia. Its ascospores are inequilateral and with a germ slit that spirals

lengthwise.

KEY worpDs — ascomycetes, Xylariaceae, decaying wood, fallen branches

Introduction

Fungi belonging in the genus Xylaria are saprophytic, endophytic, parasitic

(Whalley 1985), or symbionts in insect nests (Hawksworth et al. 1995, Pélaez et

al. 2008). They are characterized by upright, sessile, or stipitate stromata with

cylindrical, clavate, or irregularly shaped fertile parts. Many species have been

reported from different substrata throughout the world (Rogers & Samuels

1986, Bi et al. 1993, Rogers et al. 2005, Ju & Hsieh 2007, Ju et al. 2009, Rogers et

al. 2008, Carmona et al. 2009, Hladki & Romero 2010, Fan & Lin 2011, Fournier

et al. 2011, Rogers & Ju 2012).

Thailand is considered one area containing a high percentage of Xylaria

species, and several studies have reported on their biodiversity (Carroll 1963,

Schumacher 1982, Thienhirun 1997, Ruksawong & Flegal 2001, Chansrikul et

al. 2008, Petcharat et al. 2012). As currently recognized, Xylaria is represented

228 ... Srihanant, Petcharat, & Vasilyeva

in Thailand by 42 species. One recent collection that could not be identified

as any known Xylaria species is described and illustrated herein as Xylaria

thailandica.

Materials & methods

The specimens were collected from a natural plant site in a tropical rain forest in

southern Thailand. Hand-cut sections of dried material revived in 70% ethyl alcohol

were examined microscopically. Ascospore measurements were made from squash

mounts in lactophenol and the dimensions were determined from 20 fully mature

spores. A drop of Melzer’s reagent was added to determine the amyloid iodine reaction.

The photographs of stromata were taken using a Canon 550D DLSR digital camera.

Cultures were obtained from ascospores using the single spore isolation technique and

characterized on Potato Dextrose Agar (PDA) at room temperature (28-30 °C). All the

specimens are kept at Department of Pest Management, Faculty of Natural Resources,

Prince of Songkla University, Hatyai, Songkhla, Thailand (PSU-PM).

Taxonomy

Xylaria thailandica Srihanant, Petcharat & Lar.N. Vassiljeva, sp. nov. Fie. 1

MycoBank MB 807720

Differs from other species of Xylaria by its yellow-colored flesh, ascospores with a

spiraling germ slit, and asci with an urn-shaped apical ring.

Type— Thailand, Nakorn si thammarat Province, para rubber plantation, on dead wood

and fallen branches, 26. X. 2010, Natthapach Srihanant, (holotype PSU-PM 1059).

EryMoLocy—Refers to Thailand, where this fungus was collected.

STROMATA upright, gregarious, cylindrical, unbranched or (sometimes)

branched at the stipe, with short to long acute sterile apices, 3-7 cm total length

x 0.1-0.2 cm diam, externally at first brownish, becoming dark brown, then

black; internally dark brown to black at core, but yellow around perithecia,

texture soft, surface roughened by remnants of peeling layer and short dark

brown hairs; hairs not evident in old material. Young stroma filiform, dark

brown, but dull green at the top where the fruiting bodies produce conidia

(asexual spores). STIPE short to long, filiform, with a broadened discoid base,

covered with short brown hairs. PERITHECIA completely immersed, globose

to spherical, 210-310 um high x 135-250 um diam, ostioles papillate. Asc1

1.5-2 x 1-1.5 um, apical ring urn-shaped, blue in Melzer’s. AscosporEs brown,

unicellular, inequilateral, with broadly or narrowly rounded ends, 9-14.5 x

3-4.8 um, with spiraling germ slit spore-length.

Colonies on PDA: Colonies growth 14 days at room temperature (28-30 °C)

on PDA 9 cm diam., mycelium cottony, white with black color on the surface of

the agar. Stromatic structures not forming on media.

Xylaria thailandica sp. nov. (Thailand) ... 229

Fic. 1. Xylaria thailandica (holotype). A. Mature teleomorphic stroma on dead wood. B. Stromata

surface with conspicuous perithecial mounds. C. Transverse section of a stroma with dark color at

the core. D. Ascospores in lactophenol. E. Ascospore in Melzer’s reagent with an urn-shaped apical

ring. F Ascospore in lactophenol with spiral germ slit. G. Colonies on PDA in a 5 cm petri dish at

10 days. Scale bars: A =2 cm; B, C= 1 mm;D=10 um; E=5 um; F= 10 um; G= 1 ¢m

ADDITIONAL SPECIMENS EXAMINED—THAILAND, SONGKHLA PROVINCE, Prince

of Songkla University, Khao khohong nature trail, on fallen branches 5.XII.2010,

Natthapach Srihanant (PSU-PM 1060); SURATTHANI PROVINCE, oil palm plantation,

on dead wood in 24. I. 2011, Natthapach Srihanant (PSU-PM 1061).

ComMMENTS—Xylaria thailandica differs from other species on wood (Fournier

et al. 2011, Hladki & Romero 2010, Ju et al. 2009, Rogers & Samuels 1986,

Roger et al. 2008) by its yellow flesh and ascospores with spiraling germ slits.

This species can be separated from X. longipes var. tropica F. San Martin &

J.D. Rogers (Thienhirun 1997) and X. longipes Nitschke (Rogers et al. 2008),

which have larger ascospores and asci with a differently shaped apical apparatus.

The stromatal morphology of X. thailandica is close to X. cinerea J. Fournier

& M. Stadler (Fournier et al. 2011), which differs by its larger ascospores with

a slightly sinuous germ slit.

Comparisons between X. thailandica and other species are shown in TABLE 1.

230 ... Srihanant, Petcharat, & Vasilyeva

TABLE 1. Comparison of some wood-inhabiting species of Xylaria with the new taxon

X. thailandica.

SPECIES SPORE SIZE (um) GERMINATION SLIT APICAL APPARATUS

X. thailandica 9-14.5 x 3-4.8 Spiral, full spore-length Urn-shaped

X. apiculata 16-30 x 5-9 Straight, full spore-length Minute

X. castorea 8-14 x 4.5-6.5 Straight, probably fulllength § Minute

X. cinerea 13-17 x 5-6 Straight to slightly sinuous Tubular to urn-shaped

X. grammica 12-14.5 x 4-6.5 Straight, full spore-length Rectangular

X. hypoxylon 10-15.5 x 4.5-6.5 Straight to undulate, less Wedge-shaped

than full length

X. karsticola 11-14.5 x 4.2-6 Conspicuous straight, full Tubular with a slightly

spore length flared apex

X. longipes 9-15 x 5-6 Spiral, around spore Not seen

X. longipes var. tropica 14-16.5 x 5-6 Spiral, full spore-length Quadrate to rectangular

X. multiplex 9.5-11 x 4.5-5 Straight, almost full spore Not seen

length at the ventral side

X. zealandica 22-31 x 7.5-9 Straight to undulate, slightly | Urn-shaped

less than spore length

Acknowledgments

This work was supported by the Higher Education Research Promotion and National

Research University Project of Thailand, Office of the Higher Education Commission.

The first author would like to acknowledge Department of Pest Management, Prince of

Songkla University, providing necessary facilities in this research. We are thankful to

Dr. Julia Checa (Universidad de Alcala, Madrid, Spain) and Dr. Chaninun Pornsuriya

(Prince of Songkla University, Songkhla, Thailand) for reviewing the manuscript of our

paper and for their valuable suggestions.

Literature cited

Bi ZS, Zheng GY, Li TH. 1993. The macrofungus flora of China’s Guangdong province. Chinese

University of Hong Kong, Hong Kong.

Carmona A, Fournier J, Williams C, and Piepenbring M. 2009. New records of Xylariaceae from

Panama. North American Fungi 4(3): 1-11. http://dx.doi.org/10.2509/naf2009.004.003

Carroll GC. 1963. Studies in the flora of Thailand. XXIV. Pyrenomycetes. Dansk Botanisk Arkiv

23(1): 101-113.

Chandrasrikul A, Suwanarit P, Sangwanit U, Morinaga T, Nishizawa Y, Murakami Y. 2008. Diversity

of mushroom and macrofungi in Thailand. Kasetsart University Press, Bangkok, 514 p.

Fan ZY, Lin G. 2011. Xylaria hainanensis sp. nov. (Xylariaceae) from China. Mycosystema 30(4):

526-528.

Fournier J, Fressa F, Persoh D, Stadler M. 2011. Three new Xylaria species from southwestern

Europe. Mycological Progress 10: 33-52. http://dx.doi.org/10.1007/s11557-010-0671-8

Hawksworth DL, Kirk PM, Sutton BC, Pegler DN. 1995. Dictionary of the Fungi. 8 ed. Cambridge,

Cambridge University Press. 616 p.

Xylaria thailandica sp. nov. (Thailand) ... 231

Hladki AI, Romero AI. 2010. A preliminary account of Xylaria in the Tucuman Province,

Argentina, with a key to the known species from the Northern Provinces. Fungal Diversity 42:

79-96. http://dx.doi.org/1007/s13225-009-0008-6

Ju YM, Hsieh HM. 2007. Xylaria species associated with nests of Odontotermes formosanus in

Taiwan. Mycologia 99(6): 936-957. http://dx.doi.org/10.3852/mycologia.99.6.936

Ju YM, Vasilyeva L, Akulov A. 2009. Three new Xylaria species from Russian Far East. Mycologia

101(4): 548-553. http://dx.doi.org/10.3852/08-188

Ma HX, Vasilyeva L, Li Y. 2011. A new species of Xylaria from China. Mycotaxon 116: 151-155.

http://dx.doi.org/10.5248/116.151

Petcharat V, Worrapattamasri K, Srihanant N, Seepheuak P. 2012. Mushrooms of Southern Thailand.

Department of Pest Management, Faculty of Natural Resources, Prince of Songkla University,

Thailand. 507 p.

Pélaez F, Gonzalez V, Platas G, Sanchez-Ballesteros J, Rubio V. 2008. Molecular phylogenic studies

within the family Xylariaceae based on ribosomal DNA sequences. Fungal Diversity 31:

111-134.

Rogers JD, Samuels GJ. 1986. Ascomycetes of New Zealand 8. Xylaria. New Zealand Journal of

Botany 24(4): 615-650. http://dx.doi.org/10.1080/0028825X.1986.10409947

Rogers JD, Ju YM, Lehmann J. 2005. Some Xylaria species on termite nests. Mycologia 97(4):

914-923

Rogers JD, Miller AN, Vasilyeva LN. 2008. Pyrenomycetes of the Great Smoky Mountains National

Park. VI. Kretzschmaria, Nemania, Rosellinia and Xylaria (Xylariaceae). Fungal Diversity 29:

107-116.

Rogers JD, Ju YM. 2012. The Xylariaceae of the Hawaiian Islands. North American Fungi 7(9):

1-35. http://dx.doi.org/10.2509/naf2012.007.009

Ruksawong P, Flegal TW. 2001. Thai mushroom and other fungi. National Science and Technology

Development Agency, Thailand. 268 p.

Schumacher T. 1982. Ascomycetes from Northern Thailand. Nordic Journal of Botany 2: 257-263.

Thienhirun S. 1997. A preliminary account of the Xylariaceae of Thailand. Ph.D. Thesis. Liverpool

John Moores University. UK. 355 p.

Whalley AJS. (1985). The Xylariaceae: some ecological considerations. Sydowia 38: 369-382.

MYCOTAXON

http://dx.doi.org/10.5248/130.233

Volume 130, pp. 233-236 January-March 2015

Rosellinia brunneola sp. nov. and

R. beccariana new to China

WEI LI & LIN Guo*

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing

100101, China

* CORRESPONDENCE TO: guol@im.ac.cn

ABSTRACT — A new species, Rosellinia brunneola, is described from Hainan Province, China;

and Rosellinia beccariana from Yunnan Province is described as new to China.

KEY worDs —Ascomycota, pyrenomycetous fungi, Xylariaceae, taxonomy

Rosellinia species, which are cosmopolitan in their distribution, are especially

rich in tropical and subtropical regions. 142 species have been recognized

(Petrini 2013). During mycological expeditions to tropical regions of Hainan

and Yunnan Provinces, many specimens of Rosellinia have been collected, from

which a new species and a new Chinese record are described here.

Rosellinia brunneola Wei Li bis & L. Guo, sp. nov. Fics 1-3

FUNGAL NAME FEN 570115

Differs from Rosellinia mastoidiformis by its smaller stromata and ascospores with a

straight germ slit that is as long as spore length or almost so.

Type: China, Hainan, Lingao, Juliu, alt. 250 m, on dead stems, 4.XII.2010, L. Guo 11546

(HMAS 269893, holotype).

Erymo.oey: The epithet refers to the color of ascospores.

Stromata gregarious or solitary, 0.44-0.7 mm high, 0.6-0.8 mm _ wide,

semiglobose or coniform, with top tapering to a point, surface smooth, black.

Ostioles papillate. Subiculum not seen. Ectostroma 30-60 um thick, black.

Entostroma white or brownish. Asci eight-spored, with spores arranged in

biseriate manner, cylindrical, the spore-bearing part 100-120 x 10-12.5 um,

with apical ring bluing in Melzer’s iodine reagent, cylindrical, 6-7 um high,

3-4 um broad. Ascospores brownish, asymmetrically ellipsoidal, with rounded

Rosellinia brunneola sp. nov. (China) ... 235

ends, 31-39 x 5-7 um (33.8 + 3.2 x 5.9 + 1.1 um; 95%CI: 32-35 um, 5.5-6.5

um), with straight germ slit, 26-34 um long.

ComMENTs: Rosellinia brunneola is similar to R. mastoidiformis Saccas, which

differs by having larger stromata (0.5-0.9 mm high, 0.6-1 mm wide) and

ascospores with straight and indistinct germ slit that is shorter than spore

length (Petrini 2013).

Rosellinia beccariana Ces., Atti Accad. Sci. Fis. Mat. Napoli 5(21): 12. 1872.

Figs 4-6

Stromata gregarious or solitary, subglobose, sometimes with a broad, short

stipe, copper brown or black, 2.25-3.7 mm high, 2.9-3.75 mm wide, surface

smooth. Ostioles up to 600 um, pointed. Ectostroma 150-200 um thick, black.

Perithecia subglobose, 1.7-3.15 mm high, 2.6-3.5 mm wide. Asci eight-spored,

with spores arranged in biseriate manner, cylindrical, 278-355 um total length,

10-12.5(-15) um broad, the spore bearing part 201-248 um long, with apical

ring bluing in Melzer’s iodine reagent, arrowhead-shaped, 25-27 um high,

5-8 um broad. Ascospores brownish or brown, unicellular, asymmetrically

fusiform, with thread-like ends, 48-68 x 6-9 um, with straight germ slit shorter

than spore-length on flat side.

ComMMENTs: The ends of the ascospores in the Chinese material lack the slimy

sheath, which is absent in old material (Petrini 2013). The large stromata with

markedly acute ostioles make this fungus easy to recognize.

SPECIMEN EXAMINED: CHINA, YUNNAN, Mengla, Menglun, Xishuangbanna Tropical

Botanical Garden, Chinese Academy of Sciences, alt. 570 m, on corticated wood,

15.X.2013, W. Li 2903 (HMAS 253034).

Including the two species reported in this paper, 36 Rosellinia species have

been recorded in China (Teng 1963, Tai 1979, Ju & Rogers 1990, 1999, Yuan

& Zhao 1993, Liu et al. 2010, Petrini 2013).

Acknowledgements

The authors would like to express their deep thanks to Dr. L. Vasilyeva (Vladivostok,

Russia) and L.E. Petrini (Breganzona, Switzerland) for serving as pre-submission

reviewers, to Dr. Shaun Pennycook (Auckland, New Zealand) for nomenclatural review,

and to the support by the Ministry of Science and Technology of the People’s Republic

of China (No. 2013FY110400).

Fics 1-3. Rosellinia brunneola (HMAS 269893, holotype). 1. Stromata on dead stems (bar = 1 cm);

2. Ascospores; 3. Ascus. Fics 4-6. Rosellinia beccariana (HMAS 253034). 4. Stromata on wood

(bar = 1 cm); 5. Ascospores; 6. Ascus.

236 ... Li & Guo

Literature cited

Ju YM, Rogers JD. 1990. Astrocystis reconsidered. Mycologia 82: 342-349.

http://dx.doi.org/10.2307/3759905

Ju YM, Rogers JD. 1999. The Xylariaceae of Taiwan (excluding Anthostomella). Mycotaxon 73:

343-440.

Liu CE, Lu T, Gao JM, Wang MQ, Lu BS. 2010. Two new Chinese records of Rosellinia. Mycosystema

29: 459-462.

Petrini LE. 2013. Rosellinia - a world monograph. Bibliotheca Mycologica 205. 410 p.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

Yuan ZQ, Zhao ZY. 1993. Studies on the genera Amphisphaerella, Coniochaeta and Rosellinia of

Xinjiang, China. Acta Mycologica Sinica 12(3): 180-186.

MYCOTAXON

http://dx.doi.org/10.5248/130.237

Volume 130, pp. 237-239 January-March 2015

Phyllachora hainanensis sp. nov. from China

Na Lru'*, LE WANG’, Gu HUANG”, & LIN GUO?

' College of Biological Engineering, Henan University of Technology,

Lianhua Street, High & New Tech Development District, Zhengzhou 450001, China

* State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

No.1 Beichen West Road, Chaoyang District, Beijing 100101, China

* CORRESPONDENCE TO: liuna3456@163.com

ABSTRACT — A newspecies, Phyllachora hainanensis on Phyllanthus emblica (Phyllanthaceae),

causing tar spots is reported from Hainan Province, China. Phyllachora hainanensis differs

from other similar species in the size and shape of ascospores.

Key worps — Sordariomycetes, taxonomy

A new species of Phyllachora (Sordariomycetes) on leaves of Phyllanthus emblica

was collected from Hainan Province, China, in November 2007. Worldwide,

only P phyllanthophila and P. phyllanthophila var. egregia have previously been

reported from the host genus Phyllanthus (Saccardo & Saccardo 1905: 834,

Saccardo & Trotter 1913: 419).

Phyllachora hainanensis Na Liu & G. Huang, sp. nov. PLATE 1

FUNGAL NAME FN570110

Differs from Phyllachora phyllanthophila by its larger ascospores.

Type: China, Hainan Province, Xiawangling, alt. 800 m, on leaves of Phyllanthus

emblica L. (Phyllanthaceae), 13 XI 2007, Shuanghui He, L. Guo & Z.Y. Li 2013 (HMAS

245338, holotype).

EtyMoLoecy: From Hainan Province, where the new species was collected.

ASCOMATA epigenous, raised, shiny, black, sparse, subglobose, ellipsoidal

or irregular, approx. 370 x 150-260 um diam., immersed in the mesophyll,

usually with a single locule, ostiole conspicuous with a conical neck extending

through the host epidermis and cuticle, without periphyses. Asci arise from

the basal and lateral walls of the ascoma, upper wall to 50 um thick, composed

of epidermal cells occluded by melanised material. Asci 60-100 x 30-70 um,

238 ... Liu & al.

PiaTE 1. Phyllachora hainanensis (HMAS 245338, holotype).

1. Ascomata; 2-4. Asci and ascospores.

8-spored, clavate, obtuse at apex, long pedunculate, thin-walled at maturity,

unitunicate. Ascosporss biseriate or irregularly biseriate, 11-18 x 9-13 um,

ellipsoidal or subglobose, one-celled, hyaline, thin-walled, smooth, without a

gelatinous sheath. ANAMORPH not seen.

CoMMENTS—Subglobose ascospores are rare in Phyllachora. Only one other

species, P. yunnanensis Na Liu & L. Guo, collected on Lespedeza (Fabaceae)

from Yunnan Province of China, has been reported as having subglobose

(sometimes ellipsoidal) ascospores. Phyllachora yunnanensis differs from

P. hainanensis in its smaller (7-13.5 x 5-9 um) ascospores (Liu & Guo 2006).

Phyllachora hainanensis is the first Phyllachora species in China discovered

on a host species in the Phyllanthaceae. Most Phyllachora species reported

Phyllachora hainanensis sp. nov. (China) ... 239

in China occur on hosts in the Poaceae, Fabaceae, and Cyperaceae (‘Tai 1979,

Teng 1963, Lan et al. 2012, Cui 2009). The only other reports of Phyllachora

on Phyllanthus are Phyllachora phyllanthophila Henn. on Phyllanthus sp.

from Peru (Hennings 1904: 250) and P. phyllanthophila var. egregia Rehm on

Phyllanthus sp. from Brazil (Rehm 1907: 530). These taxa were subsequently

transferred to Catacauma as C. phyllanthophilum (Henn.) Theiss. & Syd. and

C. egregium (Rehm) Theiss. & Syd. (Theissen & Sydow 1915: 390-391), but

Catacauma is now synonymised with Phyllachora (von Arx & Miller 1954:

212) and P. phyllanthophila var. egregia is synonymised with P. phyllanthophila

(Index Fungorum 2015). The publications cited above provide different sets of

measurements that can be summarised as 35-60 x 12-20 um for asci, and 10-

14 x 5-9 um for ascospores; thus P. phyllanthophila differs from P. hainanensis

by its smaller asci and its narrower, ellipsoidal to subovoid ascospores.

Acknowledgments

The authors would like to thank Dr. Roger Shivas (Brisbane, Australia) and Guangyu

Sun (Northwest Agriculture and Forestry University) for serving as pre-submission

reviewers, Drs. Shuanghui He and Zhenying Li for collecting the specimen, Dr. Shaun

Pennycook (Auckland, New Zealand) for nomenclatural review, and Mr. Ziyu Cao

(Institute of Botany, Chinese Academy of Sciences) for identifying the host plants.

This study was supported by the National Natural Science Foundation of China (No.

31300015) and Natural Science Research Project of Henan Province (No. 132300410350).

Literature cited

von Arx JA, Miller E. 1951. Die Gattungen der amerosporen Pyrenomyceten. Beitrége zur

Kryptogamenflora der Schweiz 11(1). 434 p.

Cui JQ. 2009. Taxonomy and molecular phylogenetics of Colletotrichum and Phyllachora. Thesis

for Master Degree of Northwest Agriculture and Forestry University. 66 p.

Hennings P. 1904. Fungi amazonici II. a cl. Ernesto Ule collecti. Hedwigia 43: 242-273.

Index Fungorum. 2015. http://www.indexfungorum.org/names/Names.asp [accessed: February

2015].

Lan JQ, Lu HJ, Wu J., Li XL, Zhang T, Kong CC, Kang ZS. 2012. Three new records of Phyllachora

in China. Mycosystema 31: 639-641. http://dx.doi.org/10.13346/j.mycosystema.2012.04.021

Liu N, Guo L. 2006. A new species of Phyllachora (Phyllachorales) on Leguminosae from China.

Mycotaxon 97: 111-114.

Rehm H. 1907. Ascomycetes novi. Annales Mycologici 5: 516-546.

Saccardo PA, Saccardo D. 1905. Supplementum universale, pars VI. Hymenomycetae -

Laboulbeniomycetae. Sylloge Fungorum 17. 991 p.

Saccardo PA, Trotter A. 1913. Supplementum universale, pars IX. Ascomycetae - Deuteromycetae.

Sylloge Fungorum 22. 1612 p.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing, 1527 p.

Teng SC. 1963. Fungi in China. Science Press, Beijing. 808 p.

Theissen F, Sydow H. 1915. Die Dothideales. Kritisch-systematische Originaluntersuchungen.

Annales Mycologici 13: 149-746.

MY COTAXON

http://dx.doi.org/10.5248/130.241

Volume 130, pp. 241-246 January-March 2015

Two new species of Zasmidium from Nepal

RAVINDRA NATH KHARWAR’, ARCHANA SINGH’,

RAGHVENDRA SINGH ”, & SHAMBHU KUMAR?

‘Center of Advanced Study in Botany, Banaras Hindu University, Varanasi, U.P. India 221005

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

Sagar, M.P, India 470003

° Birbal Sahni Institute of Palaeobotany, 53, University Road, Lucknow, U.P, India 226007

* CORRESPONDENCE TO: drsinghtaxon@gmail.com

ABSTRACT — Zasmidium cassines on living leaves of Cassine glauca (Celastraceae) and

Z. fabaceicola on living leaves of Vigna unguiculata (Fabaceae) are described and illustrated

from a forest in the terai region of Nepal. The two new foliar fungi are compared with closely

related taxa.

Key worps — foliar diseases, hyphomycetes, morphotaxonomy

Introduction

During a 1993-94 survey of anamorphic foliar fungi in the forest of the terai

region of Nepal, two undescribed species of Zasmidium were discovered. These

are characterized by having planate conidial scars, verruculose superficial

hyphae, and solitary to rarely catenate and usually rough-walled conidia.

Verruculose superficial hyphae and rough-walled conidia are characters shared

by both Stenella and Zasmidium. Molecular analyses (Arzanlou et al. 2007)

have shown that the genus Stenella is polyphyletic and that its type species,

S. araguata Syd. (Sydow 1930), clusters within the Teratosphaeriaceae.

This species is characterized by pileate conidiogenous loci, while other

former Stenella species are distinguished by having planate Cercospora-like

conidiogenous loci and belong to the Mycosphaerellaceae. Arzanlou et al.

(2007) also demonstrated that Zasmidium cellare (Pers.) Fr. (Fries 1849), the

type species of the genus Zasmidium, clusters within the Mycosphaerellaceae

and agrees both morphologically and phylogenetically with Stenella-like

hyphomycetes now included in the that family. Hence, the name Zasmidium is

242 ... Kharwar & al.

available for all former Stenella species characterized by planate conidial scars

clustering in the Mycosphaerellaceae.

Materials & methods

Diseased leaves were collected and dried to make herbarium specimens and

deposited in Herbarium Cryptogamiae Indiae Orientalis, Indian Agricultural Research

Institute, New Delhi, India (HCIO). Isotypes are retained in the herbarium of the

Department of Botany, D.D.U. Gorakhpur University, Gorakhpur, India (GPU) for

further reference. Microscopic slides of fungal structures were prepared in lactophenol

and cotton-blue mixture from infected area of leaf. Observations were made with light

microscope using oil immersion (1000x). The figures of fungal structures were drawn

using a camera lucida and show all necessary details of morphology and ontogeny of

reproductive propagules.

Taxonomy

Zasmidium cassines Kharwar, Arch. Singh, Raghv. Singh &

Sham. Kumar, sp. nov. Fig 1

MycoBank MB 809138

Differs from Zasmidium elaeodendri by its verruculose superficial mycelium, shorter,

narrower and less septate conidia and conidiophores.

Type: Nepal, Chitwan, Narayanghat, on living leaves of Cassine glauca (Rottb.) Kuntze

[= Elaeodendron roxburghii Wight & Arn.] (Celastraceae), December 1993, coll. R.N.

Kharwar, (Holotype, HCIO 42337; isotype GPU 3002).

EryMo_oey: the epithet is derived from the name of the host genus.

Infection spots amphigenous, brown, circular to subcircular, 1-6 mm

diam., coalescing and spreading to cover most of the leaf surface. Colonies

hypophyllous, effuse. Mycelium internal and external, septate, branched,

verruculose, subhyaline to pale brown, up to 3 um wide. Stromata absent.

Conidiophores arising singly as lateral branches of superficial hyphae, semi-

macronematous, mononematous, erect to procumbent, straight to flexuous,

smooth-walled, brown, 8.5-60 um x 2-4.5 um. Conidiogenous cells integrated,

polyblastic, terminal or intercalary, sympodial, cylindrical, conidiogenous

loci cicatrized with slightly thickened scars. Conidia holoblastic, dry,

acropleurogenous, simple to catenate, rarely branched, cylindrical to obclavate,

straight to curved, subhyaline to olivaceous brown, verruculose, 0-6-septate,

sometimes constricted at septa, base obconicotruncate with slightly thickened

hilum, apex acute to subacute, 13.5-109 x 1.5-4um.

Only one species of Zasmidium, Z. elaeodendri (Kamal et al.) Kamal [= Stenella

elaeodendri Kamal et al.] has been previously reported to occur on Cassine

glauca (as Elaeodendron roxburghii; Kamal 2010). Zasmidium elaeodendri

differs by its lack of verruculose superficial mycelium, its longer and wider

Zasmidium spp. nov. (Nepal) ... 243

FIGURE 1. Zasmidium cassines (HCIO 42337)

a. Infection spots. b. Mycelium. c. Conidiophores. d. Conidia.

Scale bars:A= 20 mm, B = 20 um

branched conidiophores (2126 x 4.5-6 um), and its much longer and thicker

17-septate conidia (2132 x 3.5-7 um), which are colourless, smooth-walled,

and slightly curved with a conico-truncate base and a rounded apex. The

proposed taxon, therefore, merits description as a new species.

Some other cercosporoid hyphomycete genera have been reported on

Cassine: Pseudocercospora elaeodendri (G.P. Agarwal & Hasija) Deighton on

Cassine glauca [= Elaeodendron glaucum|] and Ps. paradoxa U. Braun & Bagyan.

on Cassine [= Elaeodendron] sp. (Deighton 1976, Braun & Bagyanarayana

244 ... Kharwar & al.

1999). Pseudocercospora differs from Z. cassines in the absence of thickened

scars at the conidial bases and on conidiogenous loci.

Kamal (2010: 260-261) listed an erroneous record of Scolecostigmina

tirumalensis on Cassine glauca [= Elaeodendron glaucum}], but this is based on a

record of Stigmella tirumalensis (Bagyanarayana et al. 1992).

1 ee,

FIGURE 2. Zasmidium fabaceicola (HCIO 42338)

a. Infection spots. b. Mycelium.c. Conidiophores.d. Conidia.

Scale bars: A = 20 mm, B = 20 um

Zasmidium fabaceicola Kharwar, Arch. Singh, Raghv. Singh &

Sham. Kumar, sp. nov. Fig 2

MycoBAnk MB 809139

Differs from Zasmidium fabacearum by its well-developed stromata, and its shorter

conidia and conidiophores.

Zasmidium spp. nov. (Nepal) ... 245

Type: Nepal, Chitwan, Narayanghat, on living leaves of Vigna unguiculata (L.) Walp.

[= Dolichos biflorus L.] (Fabaceae), January 1994, coll. R.N. Kharwar (Holotype, HCIO

42338; isotype, GPU 3413).

EryMo_oey: the epithet is derived from the host family name.

Infection spots amphigenous, blackish brown, circular to irregular, 1-15 mm

diam., later coalescing to form large irregular patches, sometimes covering

the entire leaf surface. Colonies hypophyllous, effuse, brown. Mycelium

mostly external, septate, branched, verruculose, pale brown, 1.5-3.5 um wide.

Stromata substomatal to superficial, pseudoparenchymatous, dark brown,

17-19 um in diam. Conidiophores macronematous, arising singly as lateral

branches of external hyphae, or in fascicles from stromata, erect, straight to

slightly flexuous, unbranched, smooth-walled, mid brown to dark brown below

and paler towards the apex, 4-12-septate, 75-210 x 1.5-5 um. Conidiogenous

cells integrated, polyblastic, terminal or intercalary, sympodial, cylindrical,

geniculate, scars thickened. Conidia dry, acropleurogenous, simple to catenate,

unbranched, cylindrical to obclavato-cylindrical, straight to slightly curved,

brown, verruculose, 0-3-septate, sometimes constricted at septa, hilum slightly

thickened, base obconico-truncate, apex rounded to obtuse and sometimes

cicatrized, 13.5-40 x 1.5-5um.

Only one species of Zasmidium, Z. fabacearum (K. Srivastava et al.) Kamal

(2010), has been previously reported on Dolichos, although the host was not

identified to species. Zasmidium fabacearum differs from Z. fabaceicola by

its lack of stromata and by its longer conidiophores (100-310 x 2-4 um) and

conidia (12-60 x 2-6 um).

Acknowledgments

The authors thank Prof Dr Kamal, DDU Gorakhpur University, Gorakhpur, for helpful

taxonomic advice. We are grateful to Dr. Marcin Piatek and Dr Eric H.C. McKenzie for

helpful comments and corrections. We also express our deep thanks to Dr Shaun Pennycook

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

acceptance of the manuscript for publication. Last but not least, Dr Archana Singh thanks

Department of Science and Technology (DST), Government of India, New Delhi for

providing financial support under Women Scientist Scheme (WOS-A).

Literature cited

Arzanlou M, Groenewald JZ, Gams W, Braun U, Shin HD, Crous PW. 2007. Phylogenetic and

morphotaxonomic revision of Ramichloridium and allied genera. Stud. Mycol. 58: 57-93.

http://dx.doi.org/10.3114/sim.2007.58.03

Bagyanarayana G, Braun U, Jagadeeswar P. 1992. Three new phytoparasitic fungi from India.

Mycotaxon 45: 105-108.

Braun U, Bagyanarayana G. 1999. Phytopathogenic micromycetes from India (II). Sydowia. 51(1):

1519,

246 ... Kharwar & al.

Deighton FC. 1976. Studies on Cercospora and allied genera. VI. Pseudocercospora Speg., Pantospora

Cif. and Cercoseptoria Petr. Mycol. Pap. 140. 168 p.

Fries EM. 1849. Summa Vegetabilium Scandinaviae. 2: 259-572.

Kamal. 2010. Cercosporoid fungi of India. Bishen Singh Mahendra Pal Singh, Deharadun, India

351 p.

Sydow H. 1930. Fungi Venezuelani. Ann. Mycol. 28(1-2): 29-224.

MY COTAXON

http://dx.doi.org/10.5248/130.247

Volume 130, pp. 247-251 January-March 2015

The lichen genus Leiorreuma in China

X1A0-Hua WANG” ?, L1-L1 XU’, & ZE-FENG JIA”

"College of Life Sciences, Liaocheng University, Liaocheng 252059, China

’ College of Life Sciences, Shandong Agricultural University, Taian 271018, China

*CORRESPONDENCE TO: Zfjia2008@163.com

ABSTRACT — Four species of Leiorreuma (Graphidaceae, Ostropales) are reported from China.

Leiorreuma crassimarginatum is new to science, and L. dilatatum and L. melanostalazans are

new to China. Descriptions accompany a key distinguishing the four Leiorreuma species in

China. All materials are conserved in HMAS-L.

Keywords — Ascomycota, lichenized fungi, Ostropomycetidae, taxonomy,

Introduction

According to the revised generic concept (Staiger 2002), Leiorreuma Eschw.

belongs to the family Graphidaceae (Ostropales, Ostropomycetidae, Ascomycota).

The lichen genus is characterized by apothecia with conspicuous sessile opened

lirellae, a carbonised proper exciple that is often laterally thin but basally well-

developed, an inspersed hymenium, and pale brown transversely septate or

muriform ascospores that are I-. Only one species, L. sericeum, has previously

been reported from China. We report here on three additional species for the

country, one of which we describe as new to science.

Materials & methods

The specimens examined are deposited in the Herbarium Mycologicum

Academiae Sinicae - Lichenes, Beijing, China (HMAS-L). A dissecting

microscope (TEcH XTS-30D with CANon 600D camera) anda light microscope

(OLtympus SZ-51) were used for the morphological and anatomical studies.

Measurements and illustrations were taken from the manual cross-sections of

fruit bodies in tap water. The lichen substances were detected and identified by

thin-layer chromatography (TLC) (Culberson & Kristensson 1970, Culberson

1972, Orange et al. 2001).

248 ... Wang, Xu, & Jia

Taxonomy

Leiorreuma crassimarginatum Z.F. Jia, sp. nov. Piet

FUNGAL NAME FN570068

Differs from Leiorreuma hypomelaenum by its production of norstictic acid and its

larger ascospores.

Type: China. Guizhou Province, Tongren City, Mt. Fanjing, Daling, 27°55’N 108°41’E,

alt. 1570 m, on bark, 5/X/2004, coll. Jiang-Chun Wei & Tao Zhang G425 (Holotype,

HMAS-L 071776).

ErymMo_oey: Latin crassus and marginatus, in reference to the thick thalline margin.

THALLUS corticolous, crustose, grey to pale white, rough, thin, tightly attached

to the substratum, without isidia and soralia. APoTHECTIA lirelliform, short

to elongate, open, simple or sometimes branched, 1-3 mm long, 0.3—0.6 mm

wide, sessile, with thick lateral thalline margin, often rounded at the ends, not

striate, scattered over the thallus; pisc opened, pale brown; PROPER EXCIPLE

laterally slightly and basally thickly carbonized, basal exciple 90-130 um

high; EPITHECIUM brownish, 2-5 um high; HYMENIUM brownish, inspersed,

85-130 um high, I-; HypoTHEcrum brown, 15-40 um high; paRAPHYSES

simple, 1—1.5 um wide, apices unbranched; asci cylindrical to clavate, 6-spored,

80-120 x 15—25 um; ascospores brown to brownish, thick-walled, ellipsoid,

transversely septate, often 7—12-locular, 35—45 x 10-15 um.

CHEMISTRY: C-, K+ yellow to brown, P+ yellow; norstictic and stictic acids

detected by TLC.

DISTRIBUTION & ECOLOGY: Known only from the type locality, Mt. Fanjing,

an important nature reserve in Guizhou Province, South China. Other lichens

growing in the surrounding area are species of Graphis, Diorygma, and

Ocellularia.

ComMENTs: Leiorreuma crassimarginatum is characterized by its sessile

lirellae with thick lateral thalline margins, opened discs, entire labia, thickened

carbonized basal exciples, the small, brown and thick-walled ascospores,

and the presence of norstictic and stictic acids. The new lichen is similar to

L. hypomelaenum, which differs in its smaller (22—38 x 7-11 um), 7—12-locular,

and thin-walled ascospores and its lack of norstictic acid (Staiger 2002).

Leiorreuma dilatatum (Vain.) Staiger, Biblioth. Lichenol. 85: 296 (2002).

THALLUS corticolous, crustose, pale white to grey, rough to smooth, thickish.

APOTHECIA lirelliform, short to elongate, open, simple or branched, 0.5—3 mm

long, 0.3-0.7 mm wide, sessile, with thin thalline margin; pisc opened, black

to brown; PROPER EXCIPLE laterally slightly and basally thickly carbonized,

basal exciple 30—45 um high; EPprrHEcIUM brownish, 2—5 um high; HYMENIUM

brownish, inspersed, 80-100 um high, I-; HypoTHECIUM brown, 10-20 um

L. crassimarginatum sp. nov. (Leiorreuma in China) ... 249

PLaTE.1. Leiorreuma crassimarginatum (holotype, HMAS-L 071776) A. Thallus with lirellae.

B. Apothecium, cross-section. C. Ascus containing 6 ascospores. Scales: A = 1 mm; B = 100 um;

C=50 um.

high; PARAPHYSES simple, 1—1.5 um wide, apices unbranched; asci cylindrical

to clavate, 8-spored; ascospores brown to brownish, ellipsoid, transversely

septate, often 4-locular, 20-30 x 6-8 um, I+ brown.

CHEMISTRY: C-—, K-, P-; no lichen substances detected by TLC.

DISTRIBUTION & ECOLOGY: Collected on Mt. Jianfengling, the largest

remaining tropical rainforest on Hainan Island, South China. Associated

lichens included species of Graphis, Hemithecium, and Lecanora.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Mt. Jianfengling, alt. 180 m, on

bark, 1/X/2008. Jing Li HN081374 (HMAS-L 127475).

ComMENTs: Leiorreuma dilatatum is characterized by sessile lirellae with

thin lateral thalline margins, opened discs, a basally carbonized exciple,

small brown ascospores, and the absence of lichen substances. It is similar to

250 ... Wang, Xu, & Jia

L. sericeum, which differs by its well-developed thalline and proper margins

and its smaller ascospores (14—20 x 5—8 um, Staiger 2002; 16-18 x 5.5—7.5 um,

our specimens). This is the first report of L. dilatatum from China.

Leiorreuma melanostalazans (Leight.) A.W. Archer, Telopea 11: 75 (2005).

THALLUS corticolous, crustose, pale grey, rough to smooth, thin. APOTHECIA

lirelliform, short to elongate, open, simple or branched, 1-4 mm long,

0.3-0.5 mm wide, sessile, with thin thalline margin; pisc opened, brown;

PROPER EXCIPLE inconspicuous, laterally slightly and basally thickly carbonized,

basal exciplel10—30 um high; EpIrHEcIUM brownish, 2—5 um high; HYMENIUM

brownish, inspersed, 90-110 um high, I-; HypoTHECIUM brown, 10-15 um

high; PARAPHYSES simple, 1—1.5 um wide, apices unbranched; asci cylindrical

to clavate, 8-spored; ascospores brown to brownish, ellipsoid, transversely

septate, often 7—9-locular, 25—40 x 8-10 um, I+ brown.

CHEMISTRY: C-, K+ yellow, P+ yellow; stictic acid detected by TLC.

DISTRIBUTION & ECOLOGY: Known from two localities in the tropical

rainforest in Hainan Island, South China. Associated lichens included species

of Graphis, Hemithecium, and Phaeographis.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Mt. Jianfengling, alt. 630-700

m, on bark, 1/X/2008, Jing Li HN081422 (HMAS-L 117057); 30/IX/2008, Ze-Feng Jia

HN080683 (HMAS-L 117056).

ComMENTs: Leiorreuma melanostalazans is characterized by sessile lirellae with

thin lateral thalline margins, opened discs, a basally carbonized exciple, small

brown 7-9 locular ascospores, and the presence of stictic acid. It is similar to

L. exaltatum, which differs by its lack of lichen substances (Staiger 2002). This

is the first report of L. melanostalazans from China.

Leiorreuma sericeum (Eschw.) Staiger, Biblioth. Lichenol. 85: 305 (2002).

THALLUS corticolous, crustose, pale yellow to grey, smooth, thin. APOTHECIA

lirelliform, short to elongate, open, simple or branched, 0.5-3.5 mm long,

0.2-0.3 mm wide, sessile, with thin thalline margins, pisc opened, brown

with white-grey pruina; PROPER EXCIPLE laterally thinly and basally thickly

carbonized, basal exciple 20—40 um high; EPITHECIUM brownish, 2—5 um high;

HYMENIUM brownish, inspersed, 70-90 um high, I-; HyPOTHECIUM brown,

10-15 um high; paRAPHYSES simple, 1—1.5 um wide, apices unbranched; asc1

cylindrical to clavate, 8-spored; AscosporeEs brown to brownish, ellipsoid,

transversely septate, often 4-locular, 16-18 x 5.5—7.5 um, I+ purple-brown.

CHEMISTRY: C-—, K-, P-; no lichen substances detected by TLC.

DISTRIBUTION & ECOLOGY: Distributed throughout tropical and subtropical

South China. Associated lichens included species in the Graphidaceae and

Parmeliaceae.

L. crassimarginatum sp. nov. (Leiorreuma in China) ... 251

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Mt. Jianfengling, alt. 980 m, 28/

VII/2009, Jing Li HN247-1 (HMAS-L 117025). GUANGXI PROVINCE, Shangsi County,

alt. 600 m, on bark, 31/XII/1997, Shou-Yu Guo 1638 (HMAS-L).

ComMENTs: Leiorreuma sericeum is characterized by sessile lirellae with thin

lateral thalline margins, opened discs, a laterally and basally carbonized exciple,

small (<18 um long) brown 4-locular ascospores, and the absence of lichen

substances. It is similar to L. dilatatum, which differs by its larger ascospores

and poorly developed thalline and proper margins (Staiger 2002). Leiorreuma

sericeum has previously been reported (as Phaeographis sericea) from Hong

Kong (Thrower 1988, Wei 1991).

Key to species of Leiorreuma known from China

1 Ascospores 4-locular; lichen substances absent ............. 0. eee ee ee eee eee 2

1 Ascospores >6-locular; lichen substances present ........... 0... esse eee eee eee 3

2 Thalline and proper margins poorly developed;

ASCOSPOLES ZO—SOSCGHB UME 0.2 a ine ka ged G nore dG nore 24 Hare Ea Niel L. dilatatum

2 Thalline and proper margins well developed;

ASCOSPOKes OHI Gi 6S, ors ots diets ote Sew atenat aw annie andeutleer atendthte L. sericeum

3 Norstictic and stictic acids present; ascus 6-spored;

ascospores 7—12-locular, 35-45 x 10-15 um............. L. crassimarginatum

3 Stictic acid present; ascus 8-spored;

ascospores 7—9-locular, 25—40 x 8-10 um ................ L. melanostalazans

Acknowledgments

This research was supported by the National Natural Science Foundation of China

for the funds (No. 31270066 & 31093440) and Excellent Youthful Scientist Foundation

Project of Shandong Province (No. BS2011SW028). The authors are grateful to Dr.

Santosh Joshi (CSIR-National Botanical Research Institute, Lucknow, India) and Hua-

Jie Liu (College of Life Sciences, Hebei University, China) for their valuable comments

on the manuscript and for acting as presubmission reviewers.

Literature cited

Culberson CE 1972. Improved conditions and new data for the identification of lichen products by

a standardized thin-layer chromatographic method. Journal of Chromatography 72: 113-125.

http://dx.doi.org/10.1016/0021-9673(72)80013-X

Culberson CF, Kristensson H. 1970. A standardized method for the identification of lichen products.

Journal of Chromatography 46: 85-93. http://dx.doi.org/10.1016/S0021-9673(00)83967-9

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London, UK.

Staiger B. 2002. Die Flechtenfamilie Graphidaceae. Studien in Richtung einer natiirlicheren

Gliederung. Bibliotheca Lichenologica 85. 526 p.

Thrower SL. 1988. Hong Kong lichens. Urban Council, Hong Kong. 193 p.

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

278 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.253

Volume 130, pp. 253-258 January-March 2015

Periconiella liquidambaricola sp. nov. -

a new Chinese hyphomycete

UwE BRAUN”, STEFFEN BIEN, LyDIA HONIG, & BETTINA HEUCHERT

Martin-Luther- Universitat, Institut fiir Biologie, Bereich Geobotanik und Botanischer Garten,

06099 Halle (Saale), Germany

* CORRESPONDENCE TO: uwe.braun@botanik. uni-halle.de

ABSTRACT — The new species Periconiella liquidambaricola, collected on living leaves of

Liquidambar formosana (Altingiaceae) in Jiangxi Province, China, is described, illustrated

and compared with other species of the genus Periconiella. The new fungus is the first species

described from a host belonging to the Altingiaceae. Periconiella liquidambaricola is well

characterized by its very long conidiophores and smooth, pale scolecosporous conidia. It

has similar conidia to Periconiella lygodii on fronds of Lygodium flexuosum in India, but

differs in having conidia with attenuated to acute apexes and much longer conidiophores

with uninflated bases.

KEY worps — Ascomycota, asexual morph, Periconiella qualeicola, Asia

Introduction

Leaves of Liquidambar formosana infected by a range of plant pathogenic

micromycetes were recently collected during a biodiversity-ecosystem study

in a subtropical forest near Xingangshan, Jiangxi Province, China. One of

these fungi had conidiophores with repeatedly and densely branched apices,

pigmented conidiophores with several conspicuous conidiogenous loci, and

solitary, holoblastically formed conidia that was readily identifiable as a species

of the genus Periconiella Sacc. This fungus represents the first collection of a

Periconiella species on a host in the Altingiaceae. A comparison with other

species assigned to Periconiella revealed that the Chinese collections represent

an undescribed species.

Materials & methods

Light microscopy (OLyMPus BX50) was used to examine the material (unstained

samples mounted in distilled water, oil immersion, 1000x). The ranges of sizes for

conidiophores, conidia and other structures were based on 30 measurements. Digital

254 ... Braun & al.

images were made with a Ze1ss Axioskop 2 with ZE1ss AxioCam HR and occasionally

optimized with the software ZEIss AxioVision. Freehand drawings were made on the

basis of microscopic preparations.

Taxonomy

Periconiella liquidambaricola U. Braun, S. Bien & Honig, sp. nov. FIGS. 1, 2

MycoBank MB809832

Periconiellae lygodii morphologice similis sed maculis foliorum nullis, conidiophoris

multo longioribus, ad basim non inflatis et conidiis apice attenuato vel acuto.

Type: China, Jiangxi Province, Xingangshan, subtropical forest site of the BEF-China

Project, 29.1250° N, 117.9085° E, on living and wilting leaves of Liquidambar formosana

(Altingiaceae), 8 Sep. 2013, coll. S. Bien (HOLOTYPE-HAL 2673 F).

Erymo ocy: Derived from the name of the host genus (inhabitant of Liquidambar).

LEAF spots indistinct or lacking. CoLoNtgEs on living green leaves or on

diffuse, discolored patches of wilted leaves, yellow or ochraceous to pale brown.

MycELIvo internal and external; superficial hyphae emerge through stomata,

branched, straight to sinuous, 1-4 um wide, septate, subhyaline to medium

brown, thin-walled, verruculose. STROMATA not developed. CONIDIOPHORES

solitary, scattered, arise from superficial hyphae, at the top of mother cells,

erect, straight to slightly curved, 400-700 um long, unbranched stalk 5-10 um

wide, not swollen at the base, pluriseptate throughout, distance between septa

25-50 um, dark brown throughout or somewhat paler towards the apex, wall

thickened, 1-2 um, smooth, apical portion 2-5 times densely forked, branched

part 50-100 um long, primary branches about 25-70 um long, subcylindrical.

CONIDIOGENOUS CELLS integrated, terminal, 10-30 um long and 3-6 um wide,

subcylindrical or enlarged (subclavate), up to 9 um, straight to moderately

geniculate-sinuous, with a single or several conspicuous conidiogenous loci,

Fic. 1. Periconiella liquidambaricola. Micrographs of branched apical portions of

conidiophores: A. focussed on outline; B. focussed on conidiogenous loci; C. with adherent

and detached conidia. Bars: A, C = 50 um; B = 20 um.

Periconiella liquidambaricola sp. nov. (China) ... 255

BOE HESTON ESET REY CSESRIE

Fic. 2. Periconiella liquidambaricola.

A. superficial hyphae; B. solitary conidiophores arising from superficial hyphae;

C. branched apical parts of conidiophores; D. conidia. Bars: B, C. = 50 um; D = 10 um.

256 ... Braun & al.

circular, ring-like, 1.5-3 um diam, rim darkened-refractive, barely or slightly

thickened. Conrp1A solitary, obclavate, attenuated towards the tip, 50-140 x

4-7.5 um, 3-7-septate, sometimes constricted at the septa, pale brown, thin-

walled, smooth, apex subobtuse to pointed, base short obconically truncate,

hila 2-2.5 um wide, darkened-refractive, at least around the rim.

Discussion

Arzanlou et al. (2007) published a modern description of Periconiella and

revealed its phylogenetic afhnity within the Mycosphaerellaceae. Periconiella

velutina (G. Winter) Sacc., the type species, together with most other species

assigned to Periconiella are characterized by having amero-, didymo- or

phragmosporous conidia (Ellis 1967, 1971, 1976; Priest 1991; Hosagoudar &

Braun 1995; McKenzie 1996; Arzanlou et al. 2007; Mafia et al. 2008, etc.). All

known species of this genus are specialized and confined to hosts in particular

genera of plant families. Periconiella liquidambaricola is the first species

described from a host in the Altingiaceae.

The conidial shape places P. liquidambaricola in a small group of species

characterized by scolecosporous conidia. Periconiella lygodii Arch. Singh et

al. ex U. Braun (Braun 2004), described from India on Lygodium flexuosum,

is a morphologically similar species. A record of “Periconiella sp. close to

P. rapaneae M.B. Ellis” on Lygodium flexuosum from Myanmar (Thaung 2008)

seems to belong to P lygodii. Additional records of P. lygodii are known from

the Philippines (Begum et al. 2009) and Taiwan (Kirschner & Chen 2010),

in both cases on Lygodium japonicum, and from Laos on L. polystachyum

(Phengsintham et al. 2013). Periconiella lygodii is characterized by similar

smooth scolecosporous conidia, but forms distinct leaf spots, much shorter

conidiophores that are enlarged at the base, and conidia that are rounded at the

apex (Braun 2004).

Periconiella rapaneae M.B. Ellis on Rapanea in Ecuador (Ellis 1967, 1971) is

morphologically close to P. lygodii but differs in having long primary branches

(to 170 x 5-8 um). Periconiella araliacearum G.F. Laundon (Laundon 1972), P.

arcuata Arzanlou et al. (Arzanlou et al. 2007), P longispora Kamal et al. (Kamal

et al. 1979), P maianthemi R. Kirschner (Kirschner & Piepenbring 2008) and

P. santaloidis M.B. Ellis (Ellis 1967, 1971) are similar species. Periconiella

araliacearum differs in having much shorter conidiophores (to 200 um)

with sparingly branched apices, and long, verruculose conidia (to 170 um).

Periconiella longispora, described from India on Litsea chinensis, is characterized

by having long conidiophores (to 800 um) formed in fascicles, with branches up

to 175 um, and rather broad (36-180 x 4.5-9.5 um), smooth and 3-20-septate

conidia. Periconiella arcuata is distinct with its falcate, verruculose conidia,

and P. maianthemi is distinguishable by having much shorter conidiophores

Periconiella liquidambaricola sp. nov. (China) ... 257

(200-435 wm) and shorter verruculose conidia, (13.5-)19-43(-75) um.

Periconiella santaloidis differs in having conidiophores with densely branched

apices, short branchlets and verruculose conidia.

An additional comparable species is:

Periconiella qualeicola Dorn.-Silva & Dianese, nom. nov.

MycoBank MB809833

= Periconiella longispora Dorn.-Silva & Dianese, Mycologia 95: 1246, 2004 [“2003”],

nom. illeg.,

non P. longispora Kamal, Suj. Singh & R.P. Singh,1979.

This leaf-spotting hyphomycete described from Brazil (Dornelo-Silva &

Dianese 2004), can be distinguished by its much wider cylindrical conidia

(37-95 x 5-8 um).

Periconiella liquidambaricola seems to be rather common in the subtropical

forest site near Xingangshan, Jiangxi Province. Numerous leaf samples of

Liquidambar formosana were collected for mycological examination. Mycelium

with pigmented verruculose hyphae assignable to P liquidambaricola has been

found on most leaves, but well developed conidiophores have been observed on

a limited number of leaves.

Acknowledgments

We gratefully acknowledge funding by the German Research Foundation (DFG, FOR

891/1 and 891/2) and the National Science Foundation of China (NSFC, 30710103907

and 30930005). We are also much obliged to V.A. Mel'nik (Russia) and R.G. Shivas

(Australia) for pre-submission reviews.

Literature cited

Arzanlou M, Groenewald JZ, Gams W, Braun U, Crous PW. 2007. Phylogenetic and

morphotaxonomic revision of Ramichloridium and allied genera. Studies in Mycology 58: 57-

93. http://dx.doi.org/10.3114/sim2007.58.03

Begum MM, Shivas RG, Cumagun CJR. 2009. First record of Periconiella lygodii occurring on

Lygodium japonicum in the Philippines. Australasian Plant Pathology Notes 4: 17-18.

Braun U. 2004. Periconiella species occurring on ferns. Feddes Repertorium 115: 50-55.

http://dx.doi.org/10.2307/3761924

Dornelo-Silva D, Dianese JC. 2004. Hyphomycetes on the Vochysiaceae from the Brazilian cerrado.

Mycologia 95: 1239-1251. http://dx.doi.org/10.2307/3761924

Ellis MB. 1967: Dematiaceous hyphomycetes VIII. Mycological Papers 111: 1-46.

Ellis MB. 1971: Dematiaceous Hyphomycetes. Kew.

Ellis MB. 1976: More Dematiaceous Hyphomycetes. Kew.

Hosagoudar VB, Braun U. 1995. Two new Indian hyphomycetes. Indian Phytopathology 48:

260-262.

Kamal, Singh S, Singh RP. 1979. A new species of Periconiella from India. Current Science 48:

211-212.

Kirschner R, Chen CJ. 2010. Periconiella species (anamorphic Mycosphaerellaceae) from Taiwan.

Fungal Diversity 44: 135-148. http://dx.doi.org/10.1007/s13225-010-0054-0

258 ... Braun & al.

Kirschner R, Piepenbring M. 2008. Two new hyphomycetes from Panama. Mycological Progress 7:

21-29. http://dx.doi.org/10.1007/s11557-007-0549-6

Laundon GE. 1972. Records of fungal plant diseases in New Zealand - 2. New Zealand Journal of

Botany 9: 610-624. http://dx.doi.org/10.1080/0028825X.1971.10430224

Mafia RG, Ferreira EM, Ferreira FA, Braun U, Pereira OL. 2008. Periconiella lecythidis sp. nov.,

the causal agent of a zonate leaf spot disease of the Brazilian tree Lecythis pisonis. Mycological

Progress 7: 49-52. http://dx.doi.org/10.1006/s11557-007-0552-y

McKenzie EHC. 1996. A new species of Periconiella on Pomaderris. Mycotaxon 59: 43-45.

Phengsintham P, Chukeatirote E, McKenzie EHC, Hyde KD, Braun U. 2013. Monograph of

cercosporoid fungi from Laos. Current Research in Environmental and Applied Mycology 3:

24-158.

Priest MJ. 1991: Species of Periconiella and Stenella on Proteaceae in Eastern Australia. Mycological

Research 95: 924-927. http://dx.doi.org/10.1016/S0953-7562(09)

Thaung MM. 2008. A list of Hyphales (and Agonomycetes) in Burma. Australasian Mycologist 27:

149-172.

MY COTAXON

http://dx.doi.org/10.5248/130.259

Volume 130, pp. 259-264 January-March 2015

First record of Erysiphe syringae-japonicae in Turkey

ILGAZ AKATA* & VASYL P. HELUTA

‘Ankara University, Science Faculty, Department of Biology,

TR 06100, Ankara, Turkey

°M.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine,

2 Tereshchenkivska St., Kiev, 01601, Ukraine

‘CORRESPONDING AUTHOR: fungus@hotmail.com.tr

ABSTRACT — Erysiphe syringae-japonicae was reported on leaves of Syringa vulgaris for the

first time from Turkey. A short description, distribution, and illustrations for this powdery

mildew fungus are provided and discussed briefly.

Key worps — Asia Minor, Erysiphales, invasive species, lilac, Microsphaera

Introduction

A powdery mildew collected in Japan on the lilac, Syringa amurensis var.

japonica [= S. reticulata], was described by Braun (1982) as Microsphaera

syringae-japonicae (Erysiphales, Ascomycota). Later, this species was reported

from the Russian Far East (Bunkina 1991, as “Microsphaera syringae”) and from

Korea (Shin 2000). Microsphaera syringae-japonicae was already known on

lilacs in North America and Europe, and was distinguished from M. syringae,

mainly by its evanescent mycelium, its larger number of spores in the ascus,

and its usually more extensively pigmented chasmothecial appendage bases.

In 1988, one of the authors (VP Heluta) critically examined the type specimens

of powdery mildews described from the Russian Far East, and showed that

one of them, the type specimen of M. aceris on leaves of Acer barbinerve, had

chasmothecia identical to those of M. syringae-japonicae. However they were in

adherent groups that could have drifted from another host such as a Syringa sp.

Thus, M. aceris is an earlier synonym for M. syringae-japonicae and has priority

within the genus Microsphaera Lév. However, as already discussed by Braun et

al. (2003) and Braun & Cook (2012), Microsphaera is now considered a section

of the genus Erysiphe DC.; as the epithet aceris is already occupied in Erysiphe,

the correct current name is E. syringae-japonicae |= M. syringae-japonicae].

260 ... Akata & Heluta

In the 1990s, E. syringae-japonicae from East Asia was introduced into

Europe (Seko et al. 2008, 2011), where it is now common in Germany, Poland,

Russia, Ukraine, Switzerland, etc. (Braun 1998, 2002, Piatek 2003, Heluta 2006,

Rusanov & Bulgakov 2008, Braun & Cook 2012). However, it had not been

reported in either the European or Asian parts of Turkey until autumn 2012,

when one of the authors (I. Akata) collected it on Syringa vulgaris in Ankara.

Materials & methods

Chasmothecia were studied on herbarium material dried between papers at 22-24°C.

They were examined in distilled water and photographed under a light microscope

«Primo Star» (Carl Zeiss, Germany) using the Camera «Canon A 300» and software

«AxioVision 4.7». Digital data were analysed statistically with n 230 for each character.

For study under the scanning electron microscope (JSM-6060LA, Japan), dried pieces

of infected Syringa leaves were glued onto the metal stubs and coated with gold.

Identification was made using the descriptions of Braun & Cook (2012). In addition,

the specimen was compared with samples of E. syringae-japonicae collected in Germany,

Switzerland, Ukraine, and the Russian Far East. The studied materials are deposited in

the National Herbarium of M.G. Kholodny Institute of Botany, National Academy of

Sciences of Ukraine, Kiev, Ukraine (KW).

Taxonomy

Erysiphe syringae-japonicae (U. Braun) U. Braun & S. Takam., Schlechtendalia 4:

14. 2000. Figs 1, 2

= Microsphaera syringae-japonicae U. Braun, Mycotaxon 15: 121. 1982.

= Microsphaera aceris Bunkina, Komarovskie Chteniya 21:

82. 1974 [non Erysiphe aceris DC. 1815].

= Erysiphe acerina U. Braun & S. Takam., Schlechtendalia 4: 5. 2000.

Mycelium amphigenous, white, often effused or in patches, evanescent

to sub-persistent. Hyphae thin, up to 7 um wide. Hyphal appressoria well

developed, lobed, in opposite pairs or sometimes single. Chasmothecia brown

to dark brown, scattered to gregarious on the leaf surface, 70-90 um diam.

Peridium cells polygonal to almost rounded, 10-16 x 18-21 um. Appendages

4-12, equatorial, aseptate, stiff, usually straight to rarely curved, smooth

to rough, thick-walled, hyaline but brownish or brown at the base, 75-130

um long, apices 4-5 times regularly dichotomously branched often in three

dimensions (ie., branched part not fully flat), rather compact, sometimes

primary and secondary branches elongated and therefore apical parts somewhat

deeply forked, tips recurved. Asci 3-6, sessile or short-stalked, broadly ellipsoid

or slightly ovoid, hyaline, 40-55 x 30-40 um, (5-) 6-8-spored. Ascospores

hyaline, ellipsoid or ovoid, 16-22 x 11-14 um.

SPECIMEN EXAMINED — TURKEY, ANKARA, Tandogan, 39°56’N 32°49’E, alt. 860 m, on

Syringa vulgaris L. (Oleaceae), 17.10.2012, Ilgaz Akata (KW 40328F).

Erysiphe syringae-japonicae new for Turkey ... 261

\ A 2 i .

DOM w |

ef Xk rae Ne yf Me

Wer oN i “~M ]

RON Bt S

anamennemr Sy , |

y Sehr 4 %

y i f

., oe ry

4 > . a . on. \ eS ¥ —— SSS

pschaLy Spin = Ve 36k URS 21 S88 41 Grim

/ { S 5 , oe

Fic. 1. Erysiphe syringae-japonicae (KW 40328F) on Syringa vulgaris: a - chasmothecia on the host

plant leaf (upper surface, in reflected light); b, c - chasmothecia splitting with release of ascospores

(b) and asci (c) and with appendages colored at the base; d, e - chasmothecia with appendages

(SEM); f - surface of the peridium (SEM).

Discussion

Many Syringa species are cultivated in different countries of the northern

hemisphere as important ornamental plants. Some of them are susceptible to

powdery mildew caused by Phyllactinia fraxini (DC.) Fuss and two species of

Erysiphe sect. Microsphaera. Phyllactinia fraxini is uncommon and causes little

harm to its hosts. The other two species, E. syringae and E. syringae-japonicae,

262 ... Akata & Heluta

i> 488 164m ¢

- a -

Fic. 2. Erysiphe syringae-japonicae (KW 40328F): g - basal part of chasmothecial appendage

(SEM); h, i - apical parts of appendages (SEM); j - paired hyphal appressoria (SEM); k - asci;

1 - ascospores.

are common, and E. syringae-japonicae especially can harm the hosts. Seko et

al. (2008, 2011) studied the worldwide spread of these fungi and found the

origin of E. syringae to be North America from whence it was introduced to

Europe, East Asia, and South America. However, recently the fungus has been

supplanted by the Asian E. syringae-japonicae, particularly in Europe. The last

time E. syringae was collected in Ukraine was in 2003 (Seko et al. 2011), where

it formed only the anamorph.

Erysiphe syringae-japonicae new for Turkey ... 263

Erysiphe syringae and E. syringae-japonicae are different genetically (only

94% similarity between the ITS sequences of the two species; see Seko et al.

2008) as well as morphologically. Erysiphe syringae has typically 4-5-spored

asci and nearly colorless appendages, while the asci of E. syringae-japonicae

contain (5-)6-8 spores and its appendages are more or less pigmented from

their base up to the middle part. Thus, these species can be easily distinguished

by morphological analysis.

Lilac (Syringa vulgaris) is also cultivated in Turkey as an ornamental plant.

According to the recent literature on Turkish powdery mildews in Turkey,

only E. syringae has been previously reported on this plant (Bahcecioglu &

Yildiz 2005; Bahcecioglu et al. 2006; Kabaktepe & Bahcecioglu 2006, 2009;

Kavak 2011; Severoglu & Ozyigit 2012), and there is no record of E. syringae-

japonicae. However, our sample is identical to the specimens collected in

Europe, especially in Ukraine. Thus, the spread of E. syringae-japonicae in

Turkey needs more research and monitoring.

Acknowledgments

The authors are grateful to Uwe Braun (Germany) for kindly sending several

specimens of E. syringae-japonicae collected in West Europe and for his pre-submission

review. We thank VI Sapsay (Ukraine) for his help with scanning electron microscopy.

We also thank Uwe Braun, Roger Cook, Wieslaw Mulenko, and Shaun Pennycook for

their helpful comments and careful review of this article.

Literature cited

Braun U. 1982. Descriptions of new species and combinations in Microsphaera and Erysiphe (II).

Mycotaxon 15: 121-137.

Braun U. 1998. Neufunde echter Mehltaupilze (Erysiphales) aus der BR Deutschland.

Schlechtendalia 1: 31-40.

Braun U. 2002. Erysiphe miurae and E. syringae-japonicae — new records from Russia. Mikologiya

i Fitopatologiya 36(2): 15-16.

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildews). CBS

Biodiversity Series 11. 707 p.

Braun U, Cunnington JH, Brielmaier-Liebetanz U, Ale-Agha N, Heluta V. 2003. Miscellaneous

notes on some powdery mildew fungi. Schlechtendalia 10: 91-95.

Bahcecioglu Z, Yildiz B. 2005. A study on the microfungi of Sivas Province. Turkish Journal of

Botany 29: 23-44.

Bahcecioglu Z, Kabaktepe S, Yildiz B. 2006. Microfungi isolated from plants in Kahramanmaras

Province, Turkey. Turkish Journal of Botany 30: 419-434.

Bunkina IA. 1991. Poriadok Erysiphales. Nizshie rastenia, griby i mokhoobraznyie sovetskogo

Dal’nego Vostoka. Griby, Vol. 2. Askomitsety. Erizifalnyie, klavitsipital’nyie, gelotsial’nyie.

Nauka (Leningrad): 11-142.

Heluta VP. 2006. Boroshnystorosiani hryby (poriadok Erysiphales) Kanivs’koho pryrodnoho

zapovidnyka. Zapovidna sprava v Ukraini 12(2): 23-32.

264 ... Akata & Heluta

Kabaktepe S, Bahcecioglu Z. 2006. Microfungi identified from the flora of Ordu Province in Turkey.

Turkish Journal of Botany 30: 251-265.

Kavak H. 2011. Two new records of powdery mildews with their effectiveness on three ornamentals

in Turkey. African Journal of Agricultural Research 6(5): 1076-1079.

http://dx.doi.org/10.5897/AJAR10.992.

Piatek M. 2003. Erysiphe azaleae and Erysiphe syringae-japonicae introduced in Poland. Mycotaxon

87: 121-126.

Rusanov VA, Bulgakov TS. 2008. Muchnistorosianye griby Rostovskoi oblasti. Mikologiya i

Fitopatologiya 42(4): 314-322.

Seko Y, Bolay A, Kiss L, Heluta V, Grigaliunaite B, Takamatsu S. 2008. Molecular evidence in

support of recent migration of a powdery mildew fungus on Syringa spp. into Europe from

East Asia. Plant Pathology 57(2): 243-250. http://dx.doi.org/10.1111/j.1365-3059.2007.01775x.

Seko Y, Heluta V, Grigaliunaite B, Takamatsu S. 2011. Morphological and molecular characterization

of two ITS groups of Erysiphe (Erysiphales) occurring on Syringa and Ligustrum (Oleaceae).

Mycoscience 53(3): 174-182. http://dx.doi.org/10.1007/s10267-010-0088-x.

Severoglu Z, Ozyigit I. 2012. Powdery mildew disease in some natural and exotic plants of Istanbul,

Turkey. Pakistan Journal of Botany 44: 387-393.

Shin H.-D. 2000. Erysiphaceae of Korea. National Institute of Agricultural Science and Technology,

Suwon. 321 p.

MY COTAXON

http://dx.doi.org/10.5248/130.265

Volume 130, pp. 265-273 January-March 2015

AFLP characterization of three argentine Coprotus species

ARACELI MARCELA RAMOS’, LUIS FRANCO TADIC', NAHUEL POLICELLI’,

LAURA INES FERREYRA?’, & ISABEL ESTHER CINTO’

' Lab 9 (PROPLME-PRHIDEB-CONICET), Departamento de Biodiversidad y Biologia

Experimental, Facultad de Ciencias Exactas y Naturales,

Universidad de Buenos Aires, Ciudad Universitaria, 1428, Buenos Aires, Argentina

? Departamento de Ecologia, Genética y Evolucion. Facultad de Ciencias Exactas y Naturales,

Universidad de Buenos Aires, Ciudad Universitaria, 1428, Buenos Aires, Argentina

* CORRESPONDENCE TO: icinto@bg.fcen.uba.ar

AxsstRact — AFLP methodology was applied to characterize three Coprotus species

(C. lacteus, C. niveus, C. sexdecimsporus) so as to estimate the levels of polymorphism within

species, to analyze the phenetic relationships among them, and to contrast the AFLP findings

to those of a previous RAPD study. The high number of AFLP bands obtained with the six

assayed primers allowed us to detect intra-specific variability. The genetic variability within

species obtained using AFLP (measured in terms of percentage of polymorphic loci) was two

to three times higher than those obtained by RAPD. The phenograms generated by AFLP

markers grouped all strains of the same species into three defined clusters, and a higher

association between C. lacteus and C. sexdecimsporus was also observed. The AFLP technique

could become a powerful tool for genera such as Coprotus, in which a high intra-specific

homogeneity does not allow detection of genetic variability using other PCR-based markers.

KEY worps — Ascomycota, molecular markers, coprophilous

Introduction

The fungal genus Coprotus Korf & Kimbr. comprises coprophilous

discomycetes, which belong to the Pyronemataceae Corda. Species of this

genus are homothallic, probably due to an adaptation to the substrate, which

determines their sexual isolation (Wicklow 1981). Sexual reproduction

produces ascospores dispersed by effective discharge methods.

A homothallic mating system can be very difficult to distinguish from

clonality using some molecular markers, since both systems produce progenies

that do not show evidence of segregation for these markers and yield linkage

disequilibrium at the population level over the long term (Billiard et al. 2012).

266 ... Ramos & al.

Coprotus species have been traditionally distinguished by such cytological

and morphological characters (Kimbrough et al. 1972) as the number of

ascospores per ascus, the presence or absence of pigments in paraphyses and

excipulum, and the size and shape of asci, ascospores and sterile elements.

Different concepts have been used to define fungal species. The phenotypic

concept is the classic approach based on morphology, and Coprotus species

have traditionally been classified on cytological and morphological characters

(Kimbrough et al. 1972). However, difficulties often arise while attempting to

identify Coprotus species, as they are morphologically very similar, characters

frequently overlap, and some vary greatly according to the culture conditions

(Dokmetzian et al. 2005). A polythetic concept, which circumscribes species

based on a combination of characters (Kohn 1992, Guarro et al. 1999), could

prove helpful in characterizing the Coprotus genus.

RAPD [random amplified polymorphic DNA] analysis was performed on

Coprotus species by Ramos et al. (2008). In that study, the RAPD technique

confirmed the previous identification of strains using morphological

characters (Dokmetzian et al. 2005) and provided a greater number of species-

specific bands, but genetic variability within species (in terms of percentage

of polymorphic loci) was very low. All strains had the same geographic origin

and showed identical patterns for the six primers assayed on the three Coprotus

species studied. Therefore, it would seem that to detect variability among

strains, other PCR markers, such as AFLP, should be used.

The amplified fragment length polymorphism (AFLP) method (Vos

et al. 1995) is a DNA-based fingerprinting technique, which requires no

prior sequence information on the organism under scrutiny. This method is

used to determine the degree of similarity among isolates and offers certain

advantages over other techniques, i.e., high level of identified polymorphism,

high reproducibility, and relative technical simplicity (Briad et al. 2000). AFLP

analysis has been successfully used to estimate the amount of genetic variation

within fungal species. Given that a large number of amplicons can be screened,

AFLP is especially useful for characterizing clonal lineages and for establishing

phenetic relationships among species with little morphological differentiation.

AFLP has a clear advantage over other PCR based markers such as RAPDs, not

only in terms of reproducibility but also because more loci can be screened in

each reaction.

In our present study we applied the AFLP methodology to characterize three

Coprotus species—C. lacteus (Cooke & W. Phillips) Kimbr., C. niveus (Fuckel)

Kimbr., and C. sexdecimsporus (P. Crouan & H. Crouan) Kimbr. & Korf—so as

to estimate the levels of polymorphism within species, to analyze the phenetic

relationships among them, and to contrast the findings of our AFLP study to

those of a previous RAPD study.

Coprotus AFLP characters [Argentina] ... 267

TABLE 1. Fungal strains of Coprotus species used, with their Argentinian collection

localities and BAFC numbers

STRAIN GEOGRAPHICAL LOCATION * BAFC

C. lacteus

lacA1(11), lacA3(12), lacA6(13), lacA10(14), Agronomia, CABA 874, 1937, 1940,

lacA13(15) 1941, 1942

lacL1(16), lacL 3 17), lacL4(18), lacL6 (19) Villa Lugano, CABA 1944, 1945, 1946,

1947

C. niveus

nivBC2 (25), nivBC3 (27), nivBC4 (30) Bahia Craft, Villa La Angostura, NQ 1970, 1971, 1972

nivE1 (48), nivE2 (49) Bahia Ensenada, TF 1956, 982

nivC2 (42), nivC3 (38), nivC4 (45), Campana, PBA 1960, 1961, 1962,

nivC5 (46) 1963

nivU1 (36), nivU3 (37), nivU6 (39), Ciudad Universitaria, CABA 1964, 1965, 1966,

nivU7 (40), nivU8 (41) 1967, 1968

nivL1 (34), nivL3 (35) Villa Lugano, CABA 1973, 1974

C. sexdecimsporus

sexU 1 (6), sexU2 (7), sexU4 (8) Ciudad Universitaria, CABA 1952, 1953, 1954

sexG1 (5), sexG4 (21), sexG7 (23) Los Gigantes, C 1948, 1950, 1951

* C = Cordoba province:, CABA = Buenos Aires city; NQ = Neuqueén province;

PBA = Buenos Aires province; TF = Tierra del Fuego province.

Materials & methods

Isolation and maintenance of monosporic strains

The characters used to identify the Coprotus species were those used by Kimbrough et

al. (1972). The isolation and maintenance of monosporic strains followed the procedure

indicated by Suarez et al. (2006). Thirty-one monosporic strains, from nine geographical

locations, were used (TABLE 1). All strains were deposited in the Herbarium and Culture

Collection of the Department of Biodiversity, Faculty of Natural & Exact Sciences,

University of Buenos Aires, Argentina (BAFC).

DNA extraction

Mycelium was obtained as in Ramos et al. (2000) and ground to powder in liquid

nitrogen using a sterile pestle. Genomic DNA was extracted following Gottlieb &

Lichtwardt (2001). Quality control and quantification of genomic DNA was carried out

by agarose gel (0.8% w/v) electrophoresis and by comparison with a DNA molecular-

size standard (Lambda EcoRI/HinDII, Promega Corp.). Ethidium bromide gels were

photographed under UV light.

AFLP

The AFLP methodology was carried out on 250 ng of genomic DNA using the

AFLP® Analysis System for Microorganisms Primer Kit (Invitrogen) as described in the

instructions manual with minor modifications (Gottlieb et al. 2005). Selective primers

were combined as in TABLE 2.

268 ... Ramos & al.

TABLE 2. AFLP primers used in this study

PRIMERS SEQUENCE 5’-3’

E+ACG GAC TGC GTA CCA ATT CAC G

E+AAC GAC TGC GTA CCA ATT CAA C

E+AAG GAC TGC GTA CCA ATT CAA G

M+A GAT GAG TCC TGA GTA AA

M+G GAT GAG TCC TGA GTA AG

M+C GAT GAG TCC TGA GTA AC

M+T GAT GAG TCC TGA GTA AT

* E stands for EcoRI and M stands for Msel.

All PCR amplifications were performed in a TECHNE PROGENE thermal cycler.

Polyacrylamide gel electrophoresis conditions followed Gottlieb et al. (2005). A

30-330 bp AFLP® DNA Ladder (Invitrogen) size marker was included twice in each

electrophoresis, and the size of AFLP bands scored ranged from 90 to 330 bp. AFLP

bands were visualized using the SILVER SEQUENCE TM DNA Sequencing System

(Promega).

Statistical methods

The data were extracted as a table and marked as either present (1) or absent (0).

Monomorphic bands (bands present in all individuals of a species) were discriminated

within each species and across the entire data set. The binary matrix was analyzed

with the program NTSYS-PC version 2.02 (Rohlf 1993). The unweighted pair-group

arithmetic mean method (UPGMA) cluster analysis was performed on the simple

matching (SM) association coefhicient (Sneath & Sokal 1973), and the same program

generated the phenogram showing similarity relations. The distortion produced during

the grouping analysis was calculated using the correlation cophenetic coefficients

(r) (Sokal & Rohlf 1962) using the NTSYSPC version 2.02 (Rohlf 1993) program. A

cophenetic correlation (r) of 0.8 was considered a good fit. A three-dimensional graphic

was obtained with the principal coordinated ordination method (Gower 1966).

The percentage of polymorphic loci (P = (number of polymorphic loci/ number of

loci analyzed) x 100) for each primer combination was calculated.

Results

The analysis of the six primers that produced clear and reproducible bands

by AFLP amplification yielded a total of 926 bands that appeared consistently

in all experiments among the amplified fragments of the 31 isolates. Although

the amplified bands ranged from 50 to 550 bp, we analyzed only bands from

90 to 330 bp.

TABLE 3 describes the total number of AFLP (polymorphic + monomorphic)

bands and the percentage of polymorphic loci obtained for each selective

primer combination in the three assayed fungal species. All primers detected

polymorphic bands within species and a high percentage of polymorphic loci

Coprotus AFLP characters [Argentina] ... 269

TABLE 3. Total number of AFLP (polymorphic + monomorphic) bands detected for

each selective primer combination per Coprotus species assayed.

PRIMER SPECIES ToTaL N° oF POLYMORPHIC MONOMORPHIC P%

COMBINATION BANDS BANDS BANDS

M+T/E+ACG C. lacteus 50 16 34 32

C. niveus 27 7 20 26

C. sexdecimsporus 72 60 12 83

M+C/E+AAG C. lacteus 101 28 73 28

C. niveus 108 10 98 9

C. sexdecimsporus 163 141 22 86

M+A/E+AAG C. lacteus 75 12 63 16

C. niveus 90 38 52 42

C. sexdecimsporus 152 135 17 89

M+C/E+ACG C. lacteus 77 23 54 30

C. niveus 129 75 54 58

C. sexdecimsporus 156 144 12 92

M+G/E+ACG C. lacteus 79 26 53 33.

C. niveus 80 36 44 45

C. sexdecimsporus 100 74 26 74

M+A/E+ACG C. lacteus 74 24 50 32

C. niveus 70 14 56 20

C. sexdecimsporus 143 130 13 90

were obtained for C. sexdecimsporus and a reasonable percentage for C. niveus

and for C. lacteus.

Genetic variability within species, measured in terms of percentage of

polymorphic loci was variable: P = 87 % for C. sexdecimsporus, P= 36 % for C. niveus

and, P= 28 % for C. lacteus.

The primer group M+C/E+ACG was more effective in detecting

polymorphisms (TABLE 3).

Degree of similarity and cluster analysis

The phenogram obtained using the UPGMA method was constructed

based on Simple Matching (SM) coefficients. Little distortion occurred while

constructing this phenogram, as implied by the value of the correlation

cophenetic coefficient (r=0.991). There was no direct correlation between

molecular genotype and geographic origin.

The UPGMA phenogram (Fic. 1) grouped all strains in three defined

clusters that correspond to the three Coprotus species.

The degree of similarity among species was low. Between C. lacteus and

C. sexdecimsporus, the coefficient of similarity was S = 0.5. The group formed

270 ... Ramos & al.

C5 rls

SG3

sna}dbd] ‘D>

(a)

VBA!

snuodsw2apxas 9)

SG1

snanlu ‘>

0.73

Coefficient

FiGurE 1. UPGMA phenogram showing relationships among Coprotus lacteus, C. sexdecimsporus,

and C. niveus isolates based on the simple matching (SM) association coefficient estimated from

AFLP loci.

by the isolates identified as C. niveus clustered with the other two species with

a coefficient of similarity of S = 0.46.

The main group (Fic. 1, group Gl) comprised two subgroups (SG1 and

SG2) formed by all isolates of C. niveus. Subgroup SG1 is formed by two sets

(Cl and C2) and SG2 subgroup comprised C3 and C4 sets and one isolate

separated from the rest. A second group (Fic. 1, G2) included two subgroups

(SG3 and SG4). Subgroup SG3 is formed by two sets (C5 and C6) that included

all isolates of C. lacteus. The other subgroup (SG4) comprised C7 and C8 sets,

formed by all isolates of C. sexdecimsporus.

Intra-specific variability detected for C. lacteus and C. niveus was low. This

fact is reflected by the association coefficient: SM = 0.91 among isolates of

C. lacteus and SM = 0.92 among C. niveus isolates. Only two pairs of isolates

presented 100% of similarity (association coefficient 1.0), and they belonged to

C. lacteus. Coprotus sexdecimsporus showed a higher degree of variability than

the other species, with SM = 0.54 among all isolates of this species.

The ordination of isolates through the principal coordinated method

allowed us to recognize three groups (Fic. 2, groups 1-3) in three-dimensional

dispersion revealing nearly the same relations between isolates as the

phenogram (Fic. 1). The first group included all isolates of C. lacteus, very

closely attached in the three axes. The second set showed C. sexdecimsporus

isolates differentiated in axis 1 but very closely in the other two axes. The third

comprised isolates of C. niveus closely together in the three axes, thus revealing

a high degree of similarity.

Coprotus AFLP characters [Argentina] ... 271

G Il

C. sexdecimsporus

GIll GI

C. niveus

C. lacteus

FiGuRE 2. Three-dimensional graphic of Coprotus species obtained with the Principal Coordinates

ordination technique.

Discussion

In the present study the high number of AFLP bands obtained with the six

primers assayed allowed us to detect intra-specific variability.

Previous biochemical and RAPD studies detected a high intra-specific

homogeneity (Suarez et al. 2006, Ramos et al. 2008), which could be related

to the fact that Coprotus is homothallic and produces clonal offspring (Rayner

1994). In the present study the genetic variability within species, in terms of

percentage of polymorphic loci, was two to three times higher than those

obtained by RAPD. AFLP fingerprinting is more effective for detecting genetic

diversity in species with high intra-specific homogeneity (e.g., Coprotus),

because it generated a very large number of polymorphic loci.

Genetic diversity could be a potential indicator of the relative abundance of

sexual and asexual reproduction (Chen & McDonald 1996). Furthermore other

alternatives for genetic interaction among fungi besides sexual recombination

are heterokaryosis and mitotic recombination (Esser & Kuenen 1967). In this

272 ...Ramos & al.

sense, natural selection could lead to genetic divergence and therefore increase

genetic variability levels in the homothallic fungi as Coprotus (non-out crossing

populations). The AFLP technique allowed us to detect such intraspecific

variability.

In the phenograms generated by AFLP and RAPD markers, all strains of

the same species group into three defined clusters. Besides, both phenograms

showed very similar phenetic relationships among species.

The phenogram obtained from AFLP datasets showed a higher association

between C. lacteus and C. sexdecimsporus. This result is consistent with that

obtained by means RAPD markers. This agreement might be related to the

nature of these markers since AFLP and RAPD are usually considered neutral

markers. Sampling and genotyping the progeny within sexual structures in

natural populations should allow determining whether haploid self-mating

system actually occurs in nature (Billiard et al. 2012).

The AFLP technique could become a powerful tool for researching genera

with homothallic mating systems such as Coprotus, in which a high intra-

specific homogeneity does not allow detection of variability using other PCR-

based markers.

Acknowledgements

This study was supported by a grant from Consejo Nacional de Investigaciones

Cientificas y Técnicas (CONICET) and University of Buenos Aires, Argentina. The

authors thank Dr. Lina Bettucci (Universidad de la Republica, Montevideo, Uruguay)

and Dr. Alejandra Fazio (Universidade de Sao Paulo, Brasil) for expert presubmission

review.

Literature cited

Billiard $, Lépez-Villavicencio M, Hood ME, Giraud T. 2012. Sex, outcrossing and mating types:

unsolved questions in fungi and beyond. Journal of Evolutionary Biology 25: 1020-1038.

http://dx.doi.org/10.1111/j.1420-9101.2012.02495.x

Briard M, Le Clerc V, Grzebelus LD, Senalik D, Simon PW. 2000. Modified protocols for rapid

carrot genomic DNA extraction and AFLP ™ analysis using silver stain or radioisotopes. Plant

Molecular Biology Reporter 18: 235-241. http://dx.doi.org/10.1007/BF02823994

Chen RS, McDonald BA. 1996. Sexual reproduction plays a major role in the genetic structure of

populations of the fungus Mycosphaerella graminicola. Genetics 142: 1119-1126.

Dokmetzian DA, Ramos AM, Cinto IE, Suarez ME, Ranalli ME. 2005. Seis especies del género

Coprotus (Pyronemataceae) de Argentina estudiadas en cultivo. Hickenia 3(57): 243-252.

Esser K, Kuenen R, 1967. Genetics of Fungi. Springer-Verlag, New York.

http://dx.doi.org/10.1007/978-3-642-86814-6

Gottlieb AM, Lichtwardt RW. 2001. Molecular variation within and among species of Harpellales.

Mycologia 93: 65-80. http://dx.doi.org/10.2307/3761606

Gottlieb AM, Giberti GC, Poggio L. 2005. Molecular analyses of the genus Ilex (Aquifoliaceae) in

southern South America, evidence from AFLP and ITS sequence data. American Journal of

Botany 92: 352-369. http://dx.doi.org/10.3732/ajb.92.2.352

Coprotus AFLP characters [Argentina] ... 273

Gower JC. 1966. Some distance properties of latent root and vector methods used in multivariate

analysis. Biometrika 53: 325-338. http://dx.doi.org/10.1093/biomet/53.3-4.325

Guarro J, Gené J, Stchigel AM. 1999. Developments in fungal taxonomy. Clinical Microbiology

Reviews 12: 454-500.

Kimbrough JW, Luck-Allen ER, Cain RE. 1972. North American species of Coprotus (Thelebolaceae:

Pezizales). Can. J. Bot. 50: 957-971. http://dx.doi.org/10.1139/b72-116

Kohn LM. 1992. Developing new characters for fungal systematics: an experimental approach for

determining the rank of resolution. Mycologia 84: 139-153. http://dx.doi.org/10.2307/3760244

Majer D, Mithen R, Lewis BG, Vos P, Oliver RP. 1996. The use of AFLP fingerprinting for the

detection of genetic variation in fungi. Mycol. Res. 100: 1107-1111.

http://dx.doi.org/10.1016/S0953-7562(96)80222-X

McDonald BA, Linde C. 2002. Pathogen population genetics, evolutionary potential, and

durable resistance. Ann. Rev. Phytopathol. 40: 349-379. http://dx.doi.org/10.1146/annurev.

phyto.40.120501.101443

Ramos AM, Forchiassin F, Ranalli ME, Saidman B. 2000. Isozyme analysis of different species of the

genus Saccobolus (Ascomycetes, Pezizales). Mycotaxon, 74: 447-462

Ramos. AM, Dokmetzian DA, Ferreyra LI, Ranalli ME. 2008. First report on RAPD patterns able

to differentiate some Argentinian species of the genus Coprotus (Ascomycota). Mycotaxon, 103:

9-19.

Rayner AD. 1994. Evolutionary processes affecting adaptation to saprotrophic life styles in

ascomycete populations. 261-271, in: DL Hawksworth (ed.). Ascomycete Systematics.

Problems and Perspectives in the Nineties. NATO ASI series. Plenum Press, USA.

Rohlf JE. 1993. NTSYS-PC. Numerical taxonomy and multivariate analysis system. Version 1.8.

New York: Exeter Software Applied Biostatistics Inc.

Sneath PH, Sokal RR. 1973. Numerical taxonomy: the principles and practice of numerical

classification. San Francisco: W.H. Freeman & Co.

Suarez ME, Ranalli ME, Dokmetzian DA, Ramos AM. 2006. Characterization of three species of

the genus Coprotus (Ascomycota) by isozyme analysis. Mycotaxon 97: 257-273

Sokal RR, Rohlf JF. 1962. The comparison of dendrograms by objective methods. Taxon 11: 33-40.

http://dx.doi.org/10.2307/1217208

Vos P, Hogers R, Bleeker M, Reijans M, Lee TV, Hornes M, Friters A, Pot J, Paleman J, Kuiper

M, Zabeau M. 1995 AFLP: a new technique for DNA fingerprinting. Nucl. Acids Res. 23:

4407-4414. http://dx.doi.org/10.1093/nar/23.21.4407

Wicklow DT. 1981 The coprophilous fungal community: a mycological system for examining

ecological ideas. 47-76, in: DT Wicklow, GC Carroll (eds). The Fungal Community: its

Organization and Role in the Ecosystem. Marcel Dekker, USA.

MY COTAXON

http://dx.doi.org/10.5248/130.275

Volume 130, pp. 275-278 January-March 2015

Verticicladus hainanensis, a new aquatic hyphomycete

MING-TIAN GuoS, MIN QIAOS, JIAN- YING Li, WEI WANG, & ZE-FEN YU*

Laboratory for Conservation and Utilization of Bio-resources,

Key Laboratory for Microbial Resources of the Ministry of Education,

Yunnan University, Kunming, Yunnan, 650091, P. R. China

*CORRESPONDENCE TO: Zfyuqm@hotmail.com

ABSTRACT —Verticicladus hainanensis, isolated from decaying leaves in Jianfengling Nature

Reserve, Hainan Province, China, is described and illustrated. It is characterized by fusoid,

5-8-septate, slightly grey to dark fuliginous conidia.

Key worps —fungal diversity, morphology, freshwater fungi, taxonomy

Introduction

The fungal genus Verticicladus is characterized by rhexolytic conidial

liberation from conidiogenous cells verticillately fasciculated on short

cylindrical conidiophores (Matsushima 1993). Two species have been

described, V. amazonensis Matsush. and V. subiculifer Matsush. (Matsushima

(99351996),

China has an enormous fungal diversity occurring on dead branches, rotten

submerged wood, and leaves. Many anamorphic fungi collected in southern

China have been published recently (Zhang et al. 2009a,b,c, 2011, 2012; Ma et

al. 2010, 2011a,b, 2012a,b; Wang et al. 2011; Ren et al. 2011, 2012). A species

isolated from decaying leaves in a small river from Jianfengling Nature Reserve,

Hainan Province, China had the characteristics of the genus Verticicladus but

differed from the two described species. Here, the new species is described and

illustrated in detail.

Materials & methods

The culture was isolated from dicotyledonous leaves submerged in a river in

Hainan. The rotten leaves were scissored to several 2-4 x 2-4 cm sized fragments and

then spread on to the surface of CMA (20 g cornmeal, 18 g agar, 40 mg streptomycin,

‘Min Qiao & Ming-Tian Guo contributed equally to this work

276 ... Guo, Qiao, & al.

30 mg ampicillin, 1000 ml distilled water) medium for ten days; single conidia were

isolated using sterilized needles while viewing with a BX51 microscope, and cultivated

on CMA in Petri plates. Morphological observations were made from CMA after

incubation at 28°C for one week; pure cultures and a permanent slide were deposited

in the Herbarium of the Laboratory for Conservation and Utilization of Bio-resources,

Yunnan University, Kunming, Yunnan, P.R. China (YMF; formerly Key Laboratory of

Industrial Microbiology and Fermentation Technology of Yunnan).

Taxonomy

Verticicladus hainanensis M.T. Guo & Z.F. Yu, sp. nov. PLATE 1

MycoBank MB 809830

Differs from Verticicladus amazonensis by its shorter, fusiform conidia and from V.

subiculifer by its attenuate conidia.

Type: PR China, Hainan Province, Jianfengling Nature Reserve, Chinese Academy of

Science, 18°44’N 108°49E, elev. 682 m, in a river on submerged leaves of an unidentified

dicotyledonous plant, 23 June 2011, G. Z. Yang (holotype, YMF1.04024 [dried agar

culture]; permanent slide, YMF1.040241; ex-type cultures, YMF1.040242).

EryMo toy: hainanensis refers to the province in which the species was found.

COLONIES growing moderately on CMA, attaining about 15 mm diam. after

7 days cultivation on CMA at 28°C. Colony effuse, dark olive to fuliginous,

mycelium partly superficial, partly immersed in substratum, composed of

branched, septate, smooth hyphae. Vegetative hyphae 1.9-3.8 um wide, often

2.5-3.5 um wide, hyaline to fuliginous. Conidiophores arising laterally on

hyaline vegetative hyphae, cylindrical with the apical part rounder, 6.75-9.65

x 2.35-3.5 um, hyaline to slightly grey. Conidiogenous cells cylindrical, with

obvious truncations at the apex, 5.9-14.4 x 2.1-3.7 um, slightly grey, attaching

directly on the terminal part of hyphae, solitary or on the conidiophores in

ageregates of 3 or more. Conidia fusoid, conspicuously attenuated from the

mid-part to the apex, slightly grey to dark fuliginous, smooth at surface,

37.55-65.15 x 4.2-7.6 um, 5-8-septate, often 7-septate, thick-walled, with

rhexolytic liberation from conidiogenous cells.

Discussion

Verticicladus hainanensis fits well within the genus Verticicladus based on

its conidiogenous cells that are verticillately fasciculate on short cylindrical

conidiophores and the multiseptate conidia with rhexolytic liberation from

conidiogenous cells.

Verticicladus hainanensis is similar to V. amazonensis, which differs by its

longer, cylindrical conidia (45-95 um; Matsushima 1993); and V. subiculifer

PiatE 1. Verticicladus hainanensis (holotype, YMF1.04024). A. Conidia; B. Conidiogenous cells

and conidia, C. Conidiophores and a conidium. Scale bars: A~C = 10 um.

fos)

ie}

“2

aS)

278 ... Guo, Qiao, & al.

differs from V. hainensis by the different shape of its similar sized, cylindro-

fusiform conidia lacking any obvious apical attenuation (Matsushima 1996).

Acknowledgements

This work was jointly financed by National Basic Research Program of China

(973’Program: 2013CB127506) and National Natural Science Foundation Program

of PR China (31160008, 31260007, 31360130), Grants from the Young Academic and

Technical Leader Raising Foundation of Yunnan Province (2010CI020). We are very

grateful to Prof. X.G. Zhang and Dr. R.F. Castafieda-Ruiz for critically reviewing the

manuscript and providing helpful suggestions to improve this paper.

Literature cited

Ma IG, Ma J, Zhang YD, Zhang XG. 2010. A new species of Spadicoides from Yunnan, China.

Mycotaxon 113: 255-258. http://dx.doi.org/10.5248/113.255

Ma J, Wang Y, O’Neill NR, Zhang XG. 201la. A revision of the genus Lomaantha, with the

description of a new species. Mycologia 103: 407-410. http://dx.doi.org/10.3852/10-176

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

Neosporidesmium from Hainan, China. Mycol. Prog. 10: 157-162.

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

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

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

http://dx.doi.org/10.1007/s11557-011-0775-9

Ma LG, Ma J, Zhang YD, Zhang XG. 2012b. A new species of Corynesporella and two first records

from China. Mycotaxon 119: 83-88. http://dx.doi.org/10.5248/119.83

Matsushima T. 1993. Matsushima Mycological Memoirs, No. 7. 72 p.

Matsushima T. 1996. Matsushima Mycological Memoirs, No. 9. 30 p.

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

Ren SC, Ma J, Ma LG, Zhang YD, Zhang XG. 2012. Sativumoides and Cladosporiopsis, two new

genera of hyphomycetes from China. Mycol. Prog. 11: 443-448.

http://dx.doi.org/10.1007/s11557-011-0759-9

Wang Y, Geng Y, Ma J, Wang Q, Zhang XG. 2011. Sinomyces: a new genus of anamorphic

Pleosporaceae. Fungal Biol. 115: 188-195. http://dx.doi.org/10.1016/j.funbio.2010.12.003

Zhang K, Fu HB, Zhang XG. 2009a. Taxonomic studies of Corynespora from Hainan, China.

Mycotaxon 109: 85-93. http://dx.doi.org/10.5248/109.85

Zhang K, Fu HB, Zhang XG. 2009b. Taxonomic studies of Minimelanolocus from Yunnan, China.

Mycotaxon 109: 95-101. http://dx.doi.org/10.5248/109.95

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

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

Zhang YD, Ma J, Wang Y, Ma LG, Castafieda-Ruiz RE, Zhang XG. 2011. New species and record of

Pseudoacrodictys from southern China. Mycol. Progress 10: 261-265.

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

Zhang YD, Ma J, Ma LG, Zhang XG. 2012. Two new species of Taeniolina from southern China.

Mycol. Prog. 11: 71-74. http://dx.doi.org/10.1007/s11557-010-0729-7

MY COTAXON

http://dx.doi.org/10.5248/130.279

Volume 130, pp. 279-287 January-March 2015

Geographic distribution of Sarcoporia polyspora

and S. longitubulata sp. nov.

JOSEF VLASAK™, JOSEF VLASAK JR.,

JUHA KINNUNEN’, & VIACHESLAV SPIRIN?

‘Biol. Centre of the Academy of Sciences of the Czech Republic,

Branisovskd 31, CZ-370 05 Ceské Budéjovice, Czech Rep.

*Botanical Museum, P.O. Box 7, FI-00014 University of Helsinki, Finland

* CORRESPONDENCE TO: vlasak@umbr.cas.cz

Asstract — DNA study of Sarcoporia polyspora (= Parmastomyces transmutans) revealed

only negligible sequence differences between conifer-dwelling specimens with cartilaginous

layer in the context from USA, Brazil, Europe, and Far East Asia, but a very different sequence

from three resupinate and hardwood-bound collections without such a layer and with slightly

narrower and pale brown spores from USA and Madeira Islands. This fungus, found also

among historical USA collections of S. polyspora in the BPI herbarium, is described here as

Sarcoporia longitubulata. The phylogenetic position of Sarcoporia is discussed.

Key worps — Basidiomycota, brown rot fungi, molecular taxonomy, Parmastomyces

kravtzevianus

Introduction

Sarcoporia polyspora P. Karst. is a very distinct, brown-rot polypore with

soft, resupinate to effused-reflexed basidiomes that are white to créme at first

but turn reddish-brown after bruising or drying. It can be easily recognized

by its ellipsoid and thick-walled, dextrinoid spores, which are quite unique

in polypores. The species is rare in Europe but rather common in North

America (on Tsuga spp. and Pinus spp.) and in Asia (Vlasak & Kout 2010,

Dai 2012). The fungus was described by Karsten (1894) but Karsten’s name

was abandoned for years. Based on collections from Estonia and Siberia, it

was independently described by Parmasto (1957) as Tyromyces kravtzevianus

Bondartsev & Parmasto, and a few years later Kotlaba & Pouzar (1964)

created the genus Parmastomyces for this unique polypore. In the USA,

Overholts (1941) described this species (invalidly, without a Latin diagnosis) as

“Polyporus subcartilagineus.” Later he described still another species, Polyporus

280 ... Vlasdk & al.

transmutans Oveth., which he distinguished by the more distinctly pileate habit

and its growth on Prunus (Overholts 1952). Ryvarden & Gilbertson (1984),

who found the type of P. transmutans identical with “P. subcartilagineus” as well

as with European specimens of Parmastomyces kravtzevianus, coined the name

Parmastomyces transmutans (Overh.) Ryvarden & Gilb. for all collections with

typical spores. Niemela et al. (2005), however, studied Karsten’s type material

and re-established the oldest name, Sarcoporia polyspora, for this species.

Somewhat aberrant morphology of some recent collections from hardwood

hosts and conifers led us to question the identity of Eurasian/American and

of hardwood/conifer-inhabiting Sarcoporia. We have sampled European,

American, and Asian collections from conifers and hardwoods and also

studied the type of Polyporus transmutans from the BPI herbarium along with

several other American BPI collections determined as P. transmutans to explore

whether or not all these specimens represent the same species.

Materials & methods

Morphological study

The studied specimens are deposited in the US National Fungus collection, Beltsville,

USA (BPI), in private herbarium of J. Vlasak (JV), and in mycological herbarium of

Finnish Museum of Natural History, University of Helsinki, Finland (H). They were

examined from slides prepared in cotton blue (CB) and Melzer’s reagent (IKI) with a

Leitz Diaplan microscope (x1250 magnification). Measurements were done in cotton

blue using phase contrast illumination and oil immersion (with a subjective accuracy of

0.1 um; Miettinen et al. 2006). For presenting the size range of basidiospores, 5% of the

measurements were excluded from each end of the range and are given in parentheses.

The following abbreviations are used: L’ = basidiospore length, L = mean basidiospore

length (arithmetical average of all basidiospores), W’ = basidiospore width, W = mean

basidiospore width (arithmetical average of all basidiospores), Q = L’/W’ ratio range,

and n = the number of basidiospores measured from given number of specimens. KOH-

indicates unchanged in 5% KOH and IKI- indicates unchanged in Melzer’s reagent.

Molecular phylogeny

DNA isolation and nrDNA ITS region sequencing methods follow Vlasak & Kout

(2011). The sequences were aligned with Clustal X and manually pruned. Evolutionary

analyses were conducted in MEGA6 (Tamura et al. 2013) using “all sites” and “uniform

rates” options. Other options, such as “complete deletion” and “gamma distributed with

invariant sites,” and all their combinations provided virtually identical phylogeny.

Results

Phylogenetic analysis

Sarcoporia specimens collected on hardwoods and conifers in the USA,

Madeira Island, Czech Republic, and European and Far East Russia were

Sarcoporia longitubulata sp. nov. (Macaronesia & U.S.A.) ... 281

KC595953* S. polyspora FINLAND

KC585392* P. transmutans TYPE

KC585393* S. polyspora USA conifer

KF185094* BRAZIL

Spirin 5201 RUSSIA Pinus

Spirin 4420 RUSSIA Picea

Spirin 5422 RUSSIA Picea

Spirin 6021 RUSSIA Picea

Miettinen 14814 USA Tsuga

JV 0909/1 CZECH Picea

JV 0309/93 USA Pinus

Spirin 6018 RUSSIA Pinus

JV 1009/9A hapl1 USA Quercus

JV 1009/9A hapl2 USA Quercus

100 | JV 0809/8 USA Quercus

Kinnunen 5837 MADEIRA

JQ358796* Amylocystis lapponica

Sarcoporia polyspora 100

itubulata

Sarcoporia |

0.01

FicurE 1: Phylogenetic relationships of 15 Sarcoporia specimens inferred from ITS rRNA

sequences. Amylocystis lapponica was used as outgroup. Topology from maximum likelihood

(ML) analysis. Support values along branches from ML bootstrap (1000 replicates). Branch lengths

are drawn proportional to the number of substitutions per site. GenBank numbers with asterisk

indicate sequences retrieved from GenBank; other GenBank numbers in Specimens examined.

sequenced and compared with Sarcoporia sequences in GenBank. A total

of 14 ITS and 2 nucLSU sequences were newly generated for this study and

deposited in GenBank. The ITS dataset comprising 16 sequences resulted in

an alignment with 653 characters, of which 81 were variable and 30 parsimony

informative. Extremely high sequence conservation could be demonstrated

among all conifer-dwelling specimens from different continents, with Eurasian

specimens differing from American in only one base insertion in ITS1 and

some specimens showing two haplotypes, with and without this insertion (not

shown in Fic. 1). On the other hand, most of hardwood-dwelling specimens

showed a very different sequence with about 40 mutations in ITS region.

Similarly large sequence differences were found with the nucLSU sequence

dataset (not shown), using only two sequences from each species. The aberrant

hardwood specimens were also characterized by very thick and soft resupinate

basidiocarps with long tubes and thin subiculum without cartilaginous layer.

They are described below as a new species, S. longitubulata.

282 ... Vlasdk & al.

Taxonomy

Sarcoporia longitubulata Vlasak & Spirin, sp. nov. FIGs 2- 4

MycoBank MB 809624

Sarcoporiae polysporae similis, sed tubulis longis et sporis coloratis.

Type: USA, Pennsylvania: Phoenixville, Schuylkill Canal, Ravine Trail, log of Quercus,

2 Sep 2008, J. Vlasak JV 0809/8 (Holotype, BPI 892956; isotypes, JV, H; GenBank,

KM207860, KM207863).

ErymMo oey: longitubulata (Lat.), referring to the very long tubes found in most of the

collected specimens.

BASIDIOMES annual, resupinate, pulvinaceous, up to 20 cm long, 15 cm wide,

and 2 cm thick, strikingly soft to downy, drying brittle; margin whitish to

creme, about 1 mm broad, soft and cottony, later indistinct; pore surface at

first creme with reddish tints, distinctly reddish-brown after bruising, brown to

blackish-brown on drying, the pores circular, with thin dissepiments, 3-5 per

mm; context 1-3 mm thick, soft and cottony, persistently creme colored; tube

layer very thick, up to 20 mm, extremely brittle and shattering easily when dry,

créme at first but deep brown when dry and contrasting with creme subiculum.

Spore print brown.

HYPHAL STRUCTURE monomitic; hyphae with clamp connections, KOH-,

IKI-. Context. Hyphae arranged in subparallel bundles, mostly thin-walled,

4-10 um in diam., some hyphae with thick-walls and refractive, hyaline or pale

9806

FIGURE 2: Sarcoporia basidiospores: above - S. longitubulata (holotype);

below - S. polyspora (lectotype). Scale bar = 5 um.

Sarcoporia longitubulata sp. nov. (Macaronesia & U.S.A.) ... 283

FiGurRE 3: Sarcoporia longitubulata (JV 0809/8, holotype), underside of oak log, photo in situ.

Spore deposits below left.

brown, with cyanophilous content, strongly inflated at septa, 7-18(-22) um

in diam., occasionally with double clamps. TuBEs. Hyphae subparallel, rather

loosely arranged, thin-walled, hyaline, easily collapsing, some with cyanophilous

content, (3.5-)4-5(-6) um in diam. Cystidia absent. BAsip1osporgs thick-

walled, brownish, oblong-ellipsoid, ventral side slightly concave or rarely flat,

dextrinoid, a few spores weakly or moderately cyanophilous, (4.3-)4.4-6.2

(-7.2) x (2.3-)2.5-3.3(-3.5) um (n = 150/5), L=5.29, W = 2.88, Q = 1.78-1.93.

DISTRIBUTION & ECOLOGY. On hardwood logs (predominantly Quercus

spp.) in the US North-East (Michigan, New York, Pennsylvania, Tennessee)

and Macaronesia (Madeira Is.); causing a brown rot.

ADDITIONAL SPECIMENS EXAMINED: PORTUGAL. Maperra: Madeira Island, 3 km

W from Funchal, fallen angiosperm logs, 19 Jan 2011, Kinnunen 5837 (H, GenBank

KM207865), 5845, 5848 (H). USA. PENNSYLVANIA: Gettysburg, Big Round Top,

Quercus, 5 Sep 2010, Vlasak JV 1009/9A (JV, H, GenBank KM207861, KM207862,

KM207864). Micu1GAn: Ann Arbor, hardwood, Jul 1958, Wehmeyer (BPI 223824).

NEw York: Rye, Quercus, 30 Oct 1980, Stein (BPI 222859). TENNESSEE: Great Smoky

Mt., New Found Gap, Fagus, 9 Sep 2005, Vlasak JV 0509/103 (JV, H).

ComMeEnts. Macroscopically, S. longitubulata is very distinct, so that we felt

certain from first glance that it must be a different species (Fics 3, 4). The

brown spore deposits around the basidiocarp, the very long and soft tubes,

and the relatively thin subiculum lacking a cartilaginous layer are its most

striking morphological features. Sarcoporia polyspora produces white spore

deposits and more or less pileate basidiocarps with short tubes and a thick

subiculum containing a distinct cartilaginous layer. We noted no odor of fresh

284 ... Vlasdk & al.

Figure 4: Sarcoporia longitubulata (JV 0809/8, holotype), underside of oak log, photo in situ.

basidiocarps, but on drying, S. longitubulata smelled so repugnantly that it

was impossible to keep in the same room for more than one week, although

it was not contaminated with yeasts. The primary microscopic difference of

S. longitubulata is the pale brown coloration of the basidiospores, visible in

cotton blue even with low resolution; only a few spores are more or less stained

(slightly or moderately cyanophilous). In contrast, S. polyspora basidiospores

are exceptionally strongly cyanophilous (uniformly bright blue), so that their

natural color is totally undetectable in cotton blue. Moreover, the S. longitubulata

spores are a bit narrower than in S. polyspora (TaBLE 1) and less variable in

shape and size, being uniformly oblong-ellipsoid and slightly concave on their

ventral side (Fic. 2). The basidiospores of this type occur in S. polyspora, too,

but only as a rare variation.

Discussion

We have shown that S. polyspora is widely distributed from America to

Europe and Far East Asia. The slight morphological differences we observed—

e.g., longer spores and more pileate habit in USA collections vs. shorter spores

in Far East Asian collections—have no support in DNA sequence differences

and so should be regarded as regional morphological variation. A tropical

distribution remains uncertain, as the sequenced specimens from Brazil were

collected from Pinus logs probably imported from the USA (Baldoni et al.

2015).

All studied collections were from conifers except for BPI 844703, the type of

Polyporus transmutans (Overholts 1952), collected on Prunus serotina Ehrh. in

northwestern USA. This specimen displays strikingly large and tough pilei with

Sarcoporia longitubulata sp. nov. (Macaronesia & U.S.A.) ... 285

TABLE 1. Spore measurements of two Sarcoporia species. (Limit and mean values for

each species shown in bold.)

SPECIES / SPECIMEN Ly L W’ W Q n

S. polyspora (4.2)4.7-7.2(7.3) 5.53 (2.5)2.6-3.8(4.0) 3.10 1.79 270

Miettinen 14814 (5.0)5.1-6.3(6.7) 5.64 (2.7)2.8-3.6(4.2) 3.14 1.81 30

Spirin 4420 (4.9)5.0-6.3(6.7) 5.51 (2.7)2.8-3.2(3.3) 2.97 1.86 30

Spirin 5201 (4.2)4.7-5.7(5.8) 5.05 (2.6)2.7-3.3 3.02 1.68 30

Spirin 5422 (4.7)4.8-5.7(7.1) 5.22 (2.5)2.6-3.2(3.3) 2.93 1.79 30

Spirin 6021 (4.8)5.2-6.8(7.3) 5.70 (2.6)2.7-3.6(3.7) 3.07 1.86 30

Spirin 6018 (4.8)4.9-6.1(6.2) 5.30 (2.7)2.8-3.4(3.7) 3.11 1.71 30

Vlasak 0908/15 (4.8)5.0-6.2(6.4) 5.56 (2.7)2.8-3.7(3.9) 3.17 1.76 30

Vlasak 0309/93 (5.2)5.3-7.2(7.3) 6.27 (2.8)2.9-3.8(4.0) 3.19 1.97 30

S. longitubulata (4.3)4.4-6.2(7.2) 5.29 (2.3)2.5-3.3(3.5) 2.88 1.84 150

Vlasak 0809/8 (4.3)4.4-5.8(6.0) 5.03 (2.6)2.7-3.1(3.3) 2.82 1.79 30

Vlasak 1009/9a (4.6)4.8-6.2(6.3) 5.42 (2.3)2.5-3.2(3.3) 2.82 1.93 30

Vlasak 0509/103 (4.4)4.8-5.8(7.2) 5.30 (2.7)2.8-3.3(3.5) 2.97 1.78 30

BPI 222859 (4.8)4.9-6.2(6.7) 5.39 (2.6)2.7-3.2(3.3) 2.90 1.86 30

BPI 223824 (4.8)4.9-6.0 5.32 (2.5)2.6-3.3(3.4) 2.91 1.83 30

short tubes and thick context in which no distinct gelatinous layer is present

but scattered gelatinous streaks are present. This suggests just a very large

specimen of S. polyspora and our examination revealed no distinct microscopic

differences from other S. polyspora specimens. Moreover, the ITS sequence

of the culture derived from this specimen available in GenBank (KC585392;

Ortiz-Santana et al. 2013) is identical with other S. polyspora sequences.

When we sought for a possible existing name for S. longitubulata, Tyromyces

mollissimus Maire, described from North Africa (Maire 1945) and presently

regarded as a synonym of S. polyspora, seemed a reasonable candidate.

Unfortunately, the type material is now unavailable (Pouzar 1984, Kotiranta

1998). Nevertheless, the protologue of T’ mollissimus is very clear and leaves

no doubts about its identity: it was described as growing on Pinus halepensis

Mill. and having pileate or effused-reflexed, very soft basidiocarps with thick

contextual layer (see fig. 7b, Maire 1945: 37), and a white spore print. All these

features fit well with the current concept of S. polyspora and certainly rule out

our new species. The recently described Sarcoporia neotropica from Costa Rica

(Ryvarden 2013) has imbricate soft basidiocarps and hyaline basidiospores,

characters that preclude its identity with S. longitubulata.

Because of similar spores and some of tissue features, Ryvarden &

Melo (2014) speculated about Sarcoporia’s position in the Coniophoraceae.

Molecular phylogeny refutes this hypothesis, however. The sequences of typical

286 ... Vlasak & al.

coniophoraceous species such as Coniophora puteana (Schumach.) P. Karst.

and Serpula lacrymans (Wulfen) J. Schrot. differ greatly from Sarcoporia,

which has already been recognized as a member of “antrodia clade” within

the Polyporaceae, most closely related to Amylocystis and Auriporia (Ortiz-

Santana et al. 2013). Amylocystis lapponica (Romell) Bondartsev & Singer,

is in fact quite similar, being soft and turning reddish brown after bruising

and drying, causing a brown rot, and with some microstructures staining in

Melzer’s reagent. Bondarcevomyces taxi (Bondartsev) Parmasto, which was

once referred to Parmastomyces (= Sarcoporia in current sense) by Dai &

Niemela (1995), belongs to the Boletales (Binder & Hibbett 2006). It does share

some characters with S. longitubulata (brownish spores, strong pungent odor of

drying basidiocarps) although they indicate only superficial similarity.

ADDITIONAL MATERIAL EXAMINED: Sarcoporia polyspora. CZECH REPUBLIC.

Hluboka, Karluv Hradek, Picea abies, 2 Sep 2009 Vlasak JV 0909/1 (JV, GenBank

KM207858, KM207859), 11 Aug 2009, Vlasak JV 0908/15 (JV, PRM 915664, H).

FINLAND. Uusimaa: Inkoo, Fagervik, Sept 1893, Hisinger (H) (lectotype of S. polyspora;

Lowe 1956). RUSSIA. KHABAROVSK REG.: Khabarovsk Dist., Ulika, Pinus koraiensis, 13

Aug 2012, Spirin 5201 (H, GenBank KM207856, KM207857), Malyi Kukachan, Picea

ajanensis, 19 Aug 2012, Spirin 5422 (H, GenBank KM207852); Solnechnyi Dist., Evoron,

Picea obovata, 27 Aug 2011, Spirin 4420 (H, GenBank KM207853). NIZHNY NOVGOROD

REG.: Lukoyanov Dist., Panzelka, Pinus sylvestris, 8 Aug 2013, Spirin 6018 (H, GenBank

KM207851), Picea abies, 8 Aug 2013, Spirin 6021 (H, GenBank KM207854, KM207855).

USA. MassacuusetTts: Holden, Quinapoxet, Tsuga canadensis, 26 Sep 2011, Miettinen

14814.1 (H, GenBank KM207850). PENNSYLVANIA: Wilkes-Barre, Ricketts Glen St.

Park, Pinus, 11 Sep 2003, Vlasak JV 0309/93 (JV, H, GenBank KM207849); Haycock,

Nockamixon St. Park, Pinus strobus, 7 Aug 2004, Vlasak Jr. JV 0408/4) (JV); McKean

Co., Prunus serotina, 10 Sep 1940, Campbell & Davidson 22971 (BPI 844703, holotype

of Polyporus transmutans, GenBank KC585392). NEw York: Adirondack Park, Keene

Valley, Johns Brook Trail, conifer, 23 Sep 2005, Vlasak Jr. JV 0509/193J (JV).

Bondarcevomyces taxi. RUSSIA. KHABAROvSK REG.: Khabarovsk Dist., Bolshoi

Khekhtsir Nat Res., Picea ajanensis, 5 Sep 2013, Spirin 6674 (H, JV); Komsomol’sk Dist.,

Boktor, Larix gmelinii, 18 Aug 2013, Spirin 6141 (H, JV).

Acknowledgments

The authors thank the BPI herbarium staff for the kind loan of Sarcoporia specimens,

Dr. Otto Miettinen for sequencing the Kinnunen 5837 specimen, and Prof. Teuvo Ahti

for revising the Latin diagnosis. Valuable discussions with Jiri Kout, G. Coelho, and

Y.-C. Dai and their contributions as presubmission reviewers are acknowledged. The

research of J. Vlasak was funded by institutional support RVO: 60077344.

Literature cited

Baldoni DB, Ortiz-Santana B, Coelho G, Antoniolli ZI, Jacques RJS. 2015. Sarcoporia polyspora

(Basidiomycota, Polyporales): a rare wood-decay fungus newly recorded from South America.

Nova Hedwigia 100: 177-187.

http://dx.doi.org.ezproxy.landcareresearch.co.nz/10.1127/nova_hedwigia/2014/0218

Sarcoporia longitubulata sp. nov. (Macaronesia & U.S.A.) ... 287

Binder M, Hibbett DS. 2006. Molecular systematics and biological diversification of Boletales.

Mycologia 98(6): 971-981. http://dx.doi.org/10.3852/mycologia.98.6.971

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. http://dx.doi.org/10.1007/s10267-011-0134-3

Dai YC, Niemela T. 1995. Changbai wood-rotting fungi 4. Some species described by A.S.

Bondartsev and L.V. Lyubarsky from the Russian Far East. Annales Botanici Fennici 32(4):

AL1-226.

Karsten PA. 1894. Fragmenta mycologica XLII. Hedwigia 33: 15-16.

Kotiranta H. 1998. Parmastomyces mollissimus in North Europe. Folia Cryptog. Estonica 33: 41-47.

Kotlaba F, Pouzar Z. 1964. Staining spores of Homobasidiomycetes in cotton blue and its importance

for taxonomy. Feddes Repert. 69: 131-142.

Maire R. 1945. Etudes mycologiques. Fascicle 5. Bull. Soc. Hist. Nat. Afr. N. 36: 24-42.

Miettinen O, Niemela T, Spirin W. 2006. Northern Antrodiella species: the identity of A. semisupina,

and type studies of related taxa. Mycotaxon 96: 211-239.

Niemela T, Kinnunen J, Larsson KH, Schigel D, Larsson E. 2005. Genus revisions and the new

combinations of some North European polypores. Karstenia 45: 75-80.

Ortiz-Santana B, Lindner DL, Miettinen O, Justo A, Hibbet DS. 2013. A phylogenetic overview of

the antrodia clade (Basidiomycota, Polyporales). Mycologia 105: 1391-1411.

http://dx.doi.org/10.3852/13-051

Overholts LO. 1941. New species of Polyporaceae. Mycologia 33: 90-102.

http://dx.doi.org/10.2307/3754741

Overholts LO. 1952. New species of polypores. Mycologia 44: 224-227.

Parmasto E. 1957. Eesti seente eksikaat. Mycotheca Estonica 1: no. 1-25. Tartu. 22 p.

Pouzar Z. 1984. Notes on four European polypores. Ceska Mykol. 38: 203-204.

Ryvarden L. 2013. Studies in neotropical polypores 32. Some new species from Costa Rica. Synopsis

Fungorum 30: 33-43.

Ryvarden L, Gilbertson RL. 1984. Type studies in the Polyporaceae 15, species described by O.L.

Overholts, either alone or with J.L. Lowe. Mycotaxon 19: 137-144.

Ryvarden L, Melo I. 2014. Poroid fungi of Europe. Synopsis Fungorum 31: 1-455

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA 6: Molecular Evolutionary

Genetics Analysis version 6.0. Mol. Biol. Evol. 30: 2725-2729.

http://dx.doi.org/10.1093/molbev/mst197

Vlasak J, Kout J. 2010. Sarcoporia polyspora and Jahnoporus hirtus: two rare polypores collected in

South Bohemia, Czech Republic. Czech Mycol. 61(2): 187-195.

Vlasak J, Kout J. 2011. Tropical Trametes lactinea is widely distributed in the eastern USA.

Mycotaxon 115: 271-279. http://dx.doi.org/10.5248/115.271

MY COTAXON

http://dx.doi.org/10.5248/130.289

Volume 130, pp. 289-293 January-March 2015

Coccomyces neolitseae sp. nov. from Zhangjiajie, China

QING LI’, YU-XIA CHEN’, CHUN-TAO ZHENG’,

DAN-DAN Lu! & YING-REN LIN”

' School of Life Science &? School of Forestry & Landscape Architecture,

Anhui Agricultural University, West Changjiang Road 130, Hefei, Anhui 230036, China

*CORRESPONDENCE TO: yingrenlin@yahoo.com

Asstract — A fungus found on leaves of Neolitsea levinei from the Zhangjiajie National

Forest Park in Hunan Province, China, is described as Coccomyces neolitseae. A description,

illustration, and comments are given for this taxon. The type specimen is deposited in the

Reference Collection of Forest Fungi of Anhui Agricultural University, China (AAUF).

Key worps — Rhytismataceae, Rhytismatales, morphological characteristics, taxonomy,

Lauraceae

Introduction

Coccomyces De Not. is now a large genus in the Rhytismataceae (Kirk et al.

2008). The ascomal external shape, size, and type of opening, the ascal shape

and size, and the shape of paraphyses at the apex are regarded as the most

important taxonomic characters of Coccomyces (Johnston 1986, Lin 1998).

Index Fungorum (2015) lists 196 epithets for the genus, of which Species

Fungorum (2015) accepts 106 species in Coccomyces. They usually occur

on leaves, herbaceous stems, bark, and wood of vascular plants, especially

Ericaceae, Fagaceae, and Lauraceae (Sherwood 1980). Some species, such as

C. guizhouensis Y.R. Lin & B.E. Hu, C. ledi Rehm, C. strobi J. Reid & Cain, and

C. vilis Syd. et al., may cause economically significant plant diseases (Lin et al.

1994, Rehm 1913, Reid & Cain 1961, Cannon & Minter 1984).

Recently, another new Coccomyces species was found on fallen leaves of

Neolitsea levinei in the Zhangijiajie National Forest Park, Hunan Province,

which we describe below.

Materials & methods

The study was based on collections made in 2010 from Hunan, China. Specimens

were rehydrated in water for 15 min and ascomata and conidiomata were sliced into 10-

290 ... Li & al.

15 um thick sections using a freezing microtome. The structure of the fruiting bodies

was observed in 0.1% (w/v) cotton blue in phenol glycerin. The color of the internal

structures of the fruiting bodies was observed in water. Measurements and drawings of

asci, ascospores, and paraphyses were made from squash mounts in 5% KOH solution.

Taxonomy

Coccomyces neolitseae Qing Li & Y.R. Lin, sp. nov. Fics 1-6

MycoBAnk MB 809422

Differs from Coccomyces delta by the lack of lip cells, the existence of an excipulum, a

thicker basal stroma, paraphyses that are not enlarged at the apex, shorter and thinner

ascospores lacking an obvious gelatinous sheath, and the ascomata associating with

conidiomata.

Type: China, Hunan, Zhangjiajie National Forest Park, Jinbianxi, on fallen leaves

of Neolitsea levinei Merr. (Lauraceae), 17 August 2010, X.M. Gao & Y.R. Lin 2546

(Holotype, AAUF 68654).

Erymo oey: The specific epithet refers to the host genus.

Cotoniegs on both sides of leaves, forming irregular, yellowish-white spots

6-20 mm diam.

ZONE LINES frequent, dark-brown or black, thin, entirely or partly

surrounding the bleached areas.

ConipiomaTa on both sides of leaves, predominantly on the lower side,

scattered to clustered, sometimes with several coalescing. In surface view,

conidiomata 110-215(-250) um diam., subcircular, brown to black-brown,

with a marked perimeter line or clearly defined edge, somewhat raising the

substratum surface, discharging spores through an apical ostiole. In vertical

transverse section, conidiomata intraepidermal, more or less lens-shaped.

UPPER WALL 3.5-6.5 um thick, dark brown, composed of tiny, thin-walled

angular cells 1-1.5 um diam. BAasAL WALL 2.5-6 um thick, comprised of

(1-)2-3 layers of thick-walled angular cells 2-3 um diam. SUBCONIDIOGENOUS

LAYER 2.5-4 um thick, consisting of nearly colorless, thin-walled angular cells.

CONIDIOGENOUS CELLS and ConrIpIA not present in material available.

Ascomarta in similar positions to conidiomata on the substratum, scattered,

occasionally two coalescing in the bleached spots. In surface view, ascomata

800-1200(-1600) um diam., triangular, occasionally quadrilateral, black,

with a distinct outline, moderately raising the substratum surface,

with an obvious preformed dehiscence mechanism, opening by 3-4 radial

splits that extend nearly to the ascomatal edge to expose the light yellow-

brown hymenium. Lips absent. In median vertical transverse section, ascomata

intraepidermal. COVERING STROMA 45-55 um thick in the middle of the

laterals, becoming thinner towards the top and the edge, connecting to the basal

stroma, composed of black-brown textura angularis with thick-walled cells 6-8

Coccomyces neolitseae sp. nov. (China) ... 291

Fics 1-6. Coccomyces neolitseae (holotype, AAUF 68654) on Neolitsea levinei. 1. Habit on a leaf.

2. Detail of conidiomata, ascomata, and a zone line. 3. Ascoma in median vertical transverse

section. 4. Conidioma in vertical transverse section. 5. Portion of ascoma in median transverse

vertical section. 6. Paraphyses, asci, and ascospores.

292-0 eal.

um diam. BASAL STROMA 10-18 um thick, consisting of textura angularis with

2-3 layers of black-brown, thick-walled cells. INTERNAL MATRIX OF STROMA

existing only between covering stroma and basal stroma, composed of colorless,

thin-walled angular cells 7-12 um diam., Excipu_um well-developed, 25-35

uum wide, arising from the edge of the subhymenium, comprised of rows of

hyaline, thin-walled, septate hyphae. SUBHYMENIUM 12-20 um thick, composed

of colorless textura porrecta-intricata. PARAPHYSES hyaline, aseptate, 120-150

x 0.8-1.4 um, filiform, sinuate, not swollen, occasionally branched at the apex,

not immersed in a gel. Asci ripening sequentially, 105-120 x 8-11 um, clavate,

long-stalked, thin-walled, apex rounded to obtuse, without circumapical

thickening, J-, 8-spored. AscosporEs arranged in a fascicle, 64-75 x 1.5-1.7

um, filiform, hyaline, aseptate, gradually tapered towards the acute base, not

obviously sheathed.

HOST SPECIES, HABITAT, & DISTRIBUTION: Producing conidiomata and

ascomata on fallen leaves of Neolitsea levinei; known only from the type locality,

Hunan Province, China.

COMMENTS — Coccomyces delta (Kunze) Sacc. is closely related to C. neolitseae in

ascomal appearance and in the way it is embedded, as well as the characteristics

of the zone lines. However, C. delta differs in the lack of an association of its

ascomata with conidiomata, the absence of an excipulum, the presence of a

fringe of periphysis-like lip cells, a much thinner (5 um thick) basal stroma,

slightly enlarged paraphysial apices, short-stalked, longer (100-150 tm) asci,

and wider and longer (80-100 x 2 um) ascospores with a narrow mucous

sheath (Sherwood 1980). Coccomyces delta has been observed on a wide range

of woody plants, mostly in the Europe and Atlantic islands, but with some

records from China (Sherwood 1980; Lin et al. 2012). In our Chinese materials,

the paraphyses are septate and covered by a thin mucous coating, and the asci

and ascospores are shorter than described by Sherwood (1980).

Coccomyces limitatus (Berk. & M.A. Curtis) Sacc. is easily distinguished

from the new species by trilateral to pentagonal ascomata, a covering stroma

that does not extend as far as the basal stroma, a well-developed internal

matrix comprising loose hyphae and crystals, a wider (50 um thick) excipulum

arising from inner layers of upper wall, wider (2-3 um wide) paraphyses with

swollen upper apices, and dissimilar asci and ascospores. Its asci are 4-5.6 um,

sometimes with an apical thickening, and the ascospores are only 0.8-1 um

wide and do not taper to the base (Johnston 1986).

Coccomyces neolitseae resembles Coccomyces huangshanensis Y.R. Lin & Z.Z. Li

in the nature of the internal matrix of stroma and the color of the exposed

hymenium, but C. huangshanensis has a much thinner covering stroma (8-17

Coccomyces neolitseae sp. nov. (China) ... 293

um) covered by heavily carbonized tissue at the opening that thins gradually

towards the ascomal edge, short and hyaline periphysoids, and ascospores

sometimes with a gelatinous cap at the apex (Lin et al. 2000).

Acknowledgements

The authors are grateful to Dr J.E. Taylor (Royal Botanic Garden Edinburgh, UK)

and Dr M. Ye (Hefei University of Technology, China) for serving as pre-submission

reviewers and to the Dr L.H. Wang for the identification of host plant. This study

was supported by the National Natural Science Foundation of China (No. 31270065,

31170019).

Literature cited

Cannon PF, Minter DW. 1984. Coccomyces vilis. CMI Descr. Path. Fungi & Bact. no. 792.

Index Fungorum. 2015. www.indexfungorum.org [viewed online on 23 March 2015].

Johnston PR. 1986. Rhytismataceae in New Zealand 1. Some foliicolous species of Coccomyces de

Notaris and Propolis (Fries) Corda. New Zealand J. Bot. 24: 89-124. http://dx.doi.org/10.1080

/0028825X.1986.10409723

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

10" ed. CAB International. Wallingford. 771 p.

Lin YR. 1998. Studies on Coccomyces de Notaris and Neococcomyces gen. nov. in China. [Ph.D.

thesis; in Chinese]. Northeast Forestry University. Harbin. 98 p.

Lin YR, Liu HY, Tang YP, Hu BE. 1994. Two new species of the genus Coccomyces from China [in

Chinese]. Acta Mycol. Sin. 13: 8-12.

Lin YR, Li ZZ, Xie YS, Liang SW. 2000. Studies on the genus Coccomyces from China III [in

Chinese]. Mycosystema 19: 449-453.

Lin YR, Liu HY, Hou CL, Wang SJ, Ye M, Huang CL, Xiang Y, Yu SM. 2012. Flora Fungorum

Sinicorum, vol. 40, Rhytismatales [in Chinese]. Science Press. Beijing. 261 p.

Rehm H. 1913. Ascomycetes novi VI. Ann. Mycol. 11: 150-155.

Reid J, Cain RE. 1961. The genus Therrya. Can J. Bot. 39: 1119-1129.

http://dx.doi.org/10.1139/b61-098

Sherwood MA. 1980. Taxonomic studies in the Phacidiales: The genus Coccomyces (Rhytismataceae).

Occ. Pap. Farlow Herb. Crypt. Bot. 15: 1-120.

Species Fungorum. 2015. www.speciesfungorum.org [viewed online on 23 March 2015].

MY COTAXON

http://dx.doi.org/10.5248/130.295

Volume 130, pp. 295-297 January-March 2015

Datronia ustulatiligna sp. nov. (Agaricomycetes) from India

HARPREET KAUR, GURPREET KAUR & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147 002, India

* CORRESPONDENCE TO: harpreetkaur153@gmail.com

ABSTRACT — A new poroid species, Datronia ustulatiligna, is described on partly burned

gymnospermous wood from the state of Himachal Pradesh in India.

Key worps — Basidiomycota, Rampur, Habban

While conducting fungal forays in the Narkanda, Rampur, Rajgarh, and

Habban areas of the Shimla and Sirmaur districts in Himachal Pradesh (India),

Harpreet and Dhingra made some collections of an unknown poroid fungus

associated with partly burned trees of Pinus roxburghii and Cedrus deodara.

After comparing macroscopic and microscopic features with published

descriptions (Bakshi 1971, Ryvarden & Gilbertson 1993, Nunez & Ryvarden

2001, Bernicchia 2004, Sharma 2012), we identified the polypore as a new

species of Datronia close to D. scutellata. It is distinguished by a combination of

its association with partly burned gymnospermous wood, larger pores (2-4 per

mm), and long narrowly clavate basidia.

Datronia ustulatiligna Har. Kaur, G. Kaur & Dhingra, sp. nov. PLATE 1

MycoBank MB 810285

Differs from Datronia scutellata in having thicker basidiocarps, larger pores, and long,

narrowly clavate basidia.

Type: India, Himachal Pradesh, Shimla, 8 km from Narkanda towards Rampur, on a

partly burned tree of Pinus roxburghii Sarg., 2 August 2010, Harpreet Kaur 5609 (PUN,

holotype).

Erymo.ocy: The epithet refers to the substrate, partly burned wood.

Basidiocarps annual to biennial, pileate, effused-reflexed to sessile, solitary

to imbricate; up to 4 cm wide, 2.2 cm in radius, 2.2 cm thick near base,

applanate to ungulate to dimidiate; abhymenial surface scrupose to somewhat

hirsute, concentrically sulcate and zonate, often covered with mosses, brown

296 ... Kaur, Kaur, & Dhingra

8 12

PLATE 1. Datronia ustulatiligna (holotype). 1. Hymenophore (fresh). 2. Upper part of pileus (fresh).

3. Upper part of pileus covered with mosses (fresh). 4. Hymenophore (dry). 5. Upper part of pileus

(dry). 6. Pores (with mm scale). 7. Basidiospores. 8. Basidia. 9. Groups of crystals. 10. Generative

hyphae. 11. Cystidioles. 12. Skeletal hyphae.

Datronia ustulatiligna sp. nov. (India) ... 297

to dark brown when fresh, do not change much on drying; hymenial surface

poroid, light brown to brown when fresh, brown to dark brown on drying;

pores angular to daedaleoid, 2-4 per mm; dissepiments thick to thin, entire

to lacerate; context <2 mm thick, somewhat duplex near base, brown; pore

tubes <2.0 mm long, brown; scattered groups of crystals observed in trama;

margins acute to obtuse, regular to lobed, sterile <1 mm, concolorous. Hyphal

system dimitic. Generative hyphae <3.3 um wide, branched, septate, clamped,

thin-walled in subhymenium and thick-walled in context, subhyaline. Skeletal

hyphae <3.3 um wide, rarely branched, aseptate, thick-walled, yellowish brown.

Cystidia absent, but cystidioles present in hymenial layer, 16-37 x 4-5.3 um,

fusoid, thin-walled. Basidia 40-49 x 4-6 um, narrowly clavate, 4-sterigmate,

subhyaline, clamped at the base; sterigmata up to 3.3 um long. Basidiospores

7.3-9.8 x 2.5-4 um, ellipsoid to subcylindrical to somewhat allantoid, smooth,

subhyaline, with oily contents, thin-walled inamyloid, acyanophilous.

ADDITIONAL SPECIMENS EXAMINED: INDIA, HIMACHAL PRADESH, Shimla, about 4 km

from Narkanda towards Rampur, 2 August 2010, on a partly burned gymnospermous

stump, Dhingra 5611 (PUN); on a partly burned tree of P. roxburghii, Harpreet 5608

(PUN), Harpreet 5613 (PUN); Sirmaur, 4 km from Habban towards Rajgarh, 30 August

2011, ona partly burned tree of Cedrus deodara (D. Don) G. Don, Harpreet 5612 (PUN).

REMARKS- Datronia scutellata (Schwein.) Gilb. & Ryvarden differs from our

new species by smaller pores (4-5 per mm), shorter and wider basidia (20-30

x 7-10 um), and association with angiospermous wood.

Acknowledgements

The authors thank Head (Department of Botany, Punjabi University, Patiala) for

providing research facilities; Dr. Nils Hallenberg, Professor Emeritus (University of

Gothenburg), for peer review and expert comments; and Prof. B.M. Sharma (Department

of Plant Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer review.

Literature cited

Bakshi BL. 1971. Indian Polyporaceae (on trees and timber). Indian Council of Agricultural

Research. New Delhi. 246 p.

Bernicchia A. 2004. Polyporaceae s.1. Fungi Europaei. Edizioni Candusso, Alassio, Italia. 808 p.

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

170-522.

Ryvarden L, Gilbertson RL. 1993. European polypores, vol. 1. Abortiporus - Lindtneria. Fungiflora,

Oslo. pp. 1-387

Sharma JR. 2012. Aphyllophorales of Himalaya (Auriscalpiaceae-Tremellodendropsis). Botanical

Survey of India. 590 p.

MYCOTAXON

http://dx.doi.org/10.5248/130.299

Volume 130, pp. 299-304 January-March 2015

Three new species of Xylaria from China

Gu HuANG’”, RUISHA WANG', LIN Guo’, & NA Liu?

'State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China

College of Life Sciences, Hebei Normal University, Shijiazhuang 050016, China

* CORRESPONDENCE TO: guol@im.ac.cn

ABSTRACT —Three new species, Xylaria semiglobosa, X. sphaerica, and X. jiangsuensis, are

described. They were discovered in Yunnan, Hainan, and Jiangsu Provinces in China.

KEY worDs —Ascomycota, pyrenomycetous fungi, Xylariaceae, taxonomy

Teng (1963) described 51 species and one variety of Xylaria from China and

Tai (1979) recorded 52 species, one variety, and one form. Ju & Roger (1999)

reported 40 species and 1 variety from Taiwan. Since 1979 many new Chinese

species and records of Xylaria have been published (Li & Li 1994, Xu 1999, Ju

& Hsieh 2007, Ma et al. 2011a,b, 2012, 2013, Zhu & Guo 2011, Huang et al.

2014a,b). Three additional new species are now described as follows:

Xylaria semiglobosa G. Huang & L. Guo, sp. nov. Figs 1-3

FUNGAL NAME FN570116

Differs from Xylaria glebulosa in its larger stromata and smaller ascospores.

Type: China, Yunnan, Mengla, alt. 760 m, on wood, 17.X.2013, G. Huang, L. Guo & W.

Li 230 (HMAS 270193, holotype).

Erymo oey: The epithet refers to the stromatal shape.

Stromata gregarious or scattered, semiglobose, on narrow central connective,

3-9 mm high, 4-14 mm diam; surface black, finely reticulately cracked, with

inconspicuous perithecial mounds; interior white, becoming hollow at maturity.

Perithecia subglobose or ellipsoid, 600-900 um diam; ostioles papillate. Asci

with eight ascospores arranged in a uniseriate or partially biseriate manner,

cylindrical, overall 212-237 um long, 9-16 um broad, the spore-bearing parts

300 ... Huang & al.

122-132 um long, with an apical ring staining blue in Melzer’s iodine reagent,

urn-shaped, 9-10 um high, 5-8 um broad. Ascospores dark brown, unicellular,

ellipsoid-inequilateral, sometimes pinched at the one end, smooth, (20-)

22-25(-27) x 6-7(-9) um, with an oblique germ slit much shorter than the

spore length.

ADDITIONAL SPECIMEN EXAMINED: CHINA, Hatna. Diaoluoshan Natural Reserve, on

wood, 10.X1.2012, S.H. He 201211101 (HMAS 270192).

Comments: Xylaria semiglobosa is similar to X. glebulosa (Ces.) Y.M. Ju &

J.D. Rogers, which differs in having smaller stromata [(1.5-3) mm high, 1-3

mm diam] and larger ascospores (27-31 x 8-10 um), frequently with abruptly

pinched ends (Ju & Rogers 1999). The new species also resembles X. fraseri

M.A. Whalley et al., which differs in having whitish scales on the stromatal

surface, asci with a shorter apical ring (4.5-5 um high, 3.5-4 um broad), and

ascospores with a straight germ slit (Whalley et al. 2000). Xylaria semiglobosa

is close to X. atroglobosa Hai X. Ma et al., which differs in its asci with shorter

apical rings (4.5-5 um high, 3.5-5 um broad) and ascospores with a round

hyaline non-cellular appendage up to 6-7 x 3-4 um long (Ma et al. 2012).

Xylaria sphaerica G. Huang & L. Guo, sp. nov. Fics 4-6

FUNGAL NAME FN570117

Differs from Xylaria sicula f. major in its stromata on wood with a small acute sterile

apex and long branched protrusion near the base of the head and its slightly larger

ascospores.

Type: China, Hainan, Bawangling Natural Reserve, alt. 740 m, on wood, 12.XII.2009,

Y.F. Zhu & L. Guo 133 (HMAS 270191, holotype).

Erymo.oey: The epithet refers to the stromatal shape.

Stromata with small acute sterile apex and long branches protruding

near the base of the head, 1.3-1.5 cm total length, fertile parts spherical or

subsphaeroidal, 1-1.5 mm high, 1-2 mm diam; on tomentose hair-like stipes;

surface black, smooth, with perithecial mounds; interior white. Perithecia

subglobose or ellipsoid, 540-600 um diam; ostioles papillate. Asci with eight

ascospores arranged in uniseriate manner, cylindrical, 118-128 um total

length, 7-12 um broad, the spore-bearing parts 75-83 um long, with an apical

ring staining blue in Melzer’s iodine reagent, hat-shaped, 4-5 um high, 3-4

um broad. Ascospores brown, unicellular, ellipsoid—inequilateral, occasionally

pinched at the one end, smooth, (10.5-)12-13(-15) x 5-7 um, with a straight

germ slit the length of the spore.

Fics 1-6. Xylaria semiglobosa (HMAS 270193, holotype). 1. Stromata on wood; 2. Section of

stroma; 3. Ascus and ascospores. Xylaria sphaerica (HMAS 270191, holotype). 4. Stromata on

wood; 5. Section of stroma; 6. Asci and ascospores.

Xylaria spp. nov. (China) ... 301

302 ... Huang & al.

ComMENnts: Xylaria sphaerica is similar to X. sicula f. major Ciccar., which

differs in having stromata on fallen leaves and with a long apex, slightly smaller

ascospores (9-12 x 3-6 um), and the absence of protruding branches near the

base of the stromatal head (Ciccarone 1946).

Xylaria jiangsuensis Rui S. Wang & L. Guo, sp. nov. Fics 7-10

FUNGAL NAME FN570118

Differs from Xylaria filiformis in its stromata with branched stipes and its larger and

fusiform ascospores.

Type: China, Jiangsu: Baohuashan, on dead leaves, 10.V1I.1932, S.C. Teng 977 (HMAS

7263, holotype).

Erymo.oey: The epithet refers to Jiangsu Province, where this fungus was first collected.

Stromata filiform, with a sterile apex, 3-7.5 cm total length, 0.5-1 mm diam,

fertile parts 3-6 mm long; on hair-like, branched stipes; surface black, smooth,

with conspicuous perithecial mounds; interior white. Perithecia globose or

subglobose, 450-570 um diam; ostioles papillate. Asci with eight ascospores

arranged in uniseriate manner, cylindrical, the spore-bearing parts 104-110

uum long, 5-7 um broad, with an apical ring staining blue in Melzer’s iodine

reagent, inverted hat-shaped, 2-2.5 um high, 1-1.5 um broad. Ascospores

brownish or brown, unicellular, fusiform-inequilateral, sometimes pinched at

the one end, smooth, 16.5-20(-21.5) x 4-5(-6) um, with straight germ slit the

length of the spore.

ComMENTSs: Xylaria jiangsuensis is similar to X. filiformis (Alb. & Schwein.)

Fr., which differs in having stromata on unbranched stipes and its ellipsoid-

inequilateral and smaller ascospores (11-14 x 5-6 um; Rogers & Ju 1998).

Acknowledgements

The authors would like to express their deep thanks to Prof. Anthony J.S. Whalley

(Liverpool, UK) and Dr. L. Vasilyeva (Vladivostok, Russia) for serving as pre-submission

reviewers and to Dr. Shaun Pennycook (Auckland, New Zealand) for nomenclatural

review. This study was supported by the foundation of Project of the Knowledge

Innovation Program of the Chinese Academy of Sciences (No. KSCX2-EW-Z-9).

Literature cited

Ciccarone A. 1946. Alcune osservazioni su una forma di Xylaria sicula Pass. e Beltr. Nuovo Giorn.

Bot. Ital., n.s. 53: 356-358.

Huang G, Guo L, Liu N. 2014a. Xylaria byttneriae sp. nov. from Yunnan Province in China.

Mycosystema 33: 567-570.

Huang G, Guo L, Liu N. 2014b.Two new species of Xylaria and X. diminuta new to China.

Mycotaxon 129(1): 149-152. http://dx.doi.org/10.5248/129.149

Ju YM, Rogers JD. 1999. The Xylariaceae of Taiwan (excluding Anthostomella). Mycotaxon 73:

343-440.

Xylaria spp. nov. (China) ... 303

Fics 7-10. Xylaria jiangsuensis (HMAS 7263, holotype). 7. Stromata on dead leaf; 8. Section of

stroma; 9-10. Asci and ascospores.

Ju YM, Hsieh HM. 2007. Xylaria species associated with nests of Odontotermes formosanus in

Taiwan. Mycologia 99: 936-957. http://dx.doi.org/10.3852/mycologia.99.6.936

Li YX, Li HJ. 1994. A novel species of Xylaria. J. Nanjing Agr. Univ. 17(3): 145-147.

Ma HX, Vasilyeva LN, Li Y. 2011a. A new species of Xylaria from China. Mycotaxon 116: 151-155.

http://dx.doi.org/10.5248/116.151

Ma HX, Vasilyeva LN, Li Y. 2011b. Xylaria choui, a new species from China. Sydowia 63: 79-83.

Ma HX, Vasilyeva LN, Li Y. 2012. The genus Xylaria in the south of China - 3. X. atroglobosa sp.

nov. Mycotaxon, 119: 381-384. http://dx.doi.org/10.5248/119.381

Ma HX, Vasilyeva LN, Li Y. 2013. The genus Xylaria in the south of China - 4. X. hemisphaerica sp.

nov. from Yunnan Province. Mycosystema 32: 602-605.

304 ... Huang & al.

Rogers JD, Ju YM. 1998. Keys to the Xylariaceae (excluding Anthostomella) of the British Isles. Bot

J Scot 50: 153-160. http://dx.doi.org/10.1080/037466098086849 12

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

Whalley MA, Ju YM, Rogers JD, Whalley AJS. 2000. New xylariaceous fungi from Malaysia.

Mycotaxon 74: 135-140.

Xu AS. 1999. A new species of Xylaria. Mycosystema 18(2): 137-140

Zhu YF, Guo L. 2011. Xylaria hainanensis sp. nov. (Xylariaceae) from China. Mycosystema 30:

526-528.

MY COTAXON

http://dx.doi.org/10.5248/130.305

Volume 130, pp. 305-306 January-March 2015

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 130-1

Ambomucor ovalisporus X.Y. Liu & R.Y. Zheng, p. 166

Ambomucor seriatoinflatus var. longior X.Y. Liu & R.Y. Zheng, p. 168

Antherospora sukhomlyniae K.G. Savchenko, p. 57

Anungitea guangxiensis J.W. Xia & X.G. Zhang, p. 43

Bertia hainanensis Lar.N. Vassiljeva, H.X. Ma, Chernyshev &

S.L. Stephenson, p. 198

Cladonia pseudofissa (Asahina) Ahti, Pino-Bodas & S. Stenroos, p. 98

Coccomyces neolitseae Qing Li & Y.R. Lin, p. 290

Coccomyces prominens Y.F. Xu &Y.R. Lin, p. 74

Datronia ustulatiligna Har. Kaur, G. Kaur & Dhingra, p. 295

Dentipellicula austroafricana Jia J. Chen, L.L. Shen & Y.C. Dai, p. 20

Ellisembia longchiensis J.W. Xia & X.G. Zhang, p. 44

Fistulina subhepatica B.K. Cui & J. Song, p. 53

Hyphoderma hallenbergii Man. Kaur, Avneet P. Singh & Dhingra, p. 223

Lactarius indochrysorrheus K. Das & Verbeken, p. 110

Lactarius olivaceoglutinus K. Das & Verbeken, p. 114

Lactarius pyriodorus K. Das & Verbeken, p. 118

Lactarius yumthangensis K. Das & Verbeken, p. 124

Leiorreuma crassimarginatum Z.F. Jia, p. 248

Lophodermium heterocladum H.L. Gu & Y.R. Lin, p. 192

Perenniporia koreana Y. Jang & J.J. Kim, p. 174

Periconiella liquidambaricola U. Braun, S. Bien & Honig, p. 254

Periconiella qualeicola Dorn.-Silva & Dianese, p. 257

= Periconiella longispora Dorn.-Silva & Dianese 2004, nom. illeg.

non P. longispora Kamal, Suj. Singh & R.P. Singh 1979

Phyllachora hainanensis Na Liu & G. Huang, p. 237

Pseudoplectania lignicola Glejdura, V. Kucera, Lizon & Kunca, p. 2

Rosellinia brunneola Wei Li bis & L. Guo, p. 233

Sarcoporia longitubulata Vlasak & Spirin, p. 282