IV ... MYCOTAXON 128

MYCOTAXON

VOLUME ONE HUNDRED TWENTY-EIGHT — TABLE OF CONTENTS

COVER SECTION

PP GUG ES og 6 eR NECN 6 ORE EW PR EE oe Saas see Se UN vi

ROVTOWETS isle: Vit, FR Ee Ae Ee eet Se aie cee haat Vins Sin Aany Rint vii

SUbIRISS ON PLOCCAUT ES: whe) Bieter Rw Leva paw he eg reed heen Las Viii

EYOTA TE AIOR |. deh 9 act SR TERE Ge wie sith Meee Sate Aa okt te cn eS ix

RESEARCH ARTICLES

Tylopilus: a new species and a new record from Pakistan

S. SARWAR, A.N. KHALID, & A.R. NIAZI

Anacraspedodidymum, a new genus from submerged wood in Brazil

CAROLINA RIBEIRO SILVA, RAFAEL EF, CASTANEDA-RUIZ,

& Luis FERNANDO PaSCHOLATI GUSMAO

Two new rust species on Fabaceae from Brazil

MAuvRICIO SALAZAR YEPES & ANIBAL ALVES DE CARVALHO JUNIOR

The first record of Hebeloma vinosophyllum (Strophariaceae) in

Southeast Asia Bao-THuy QuyEN Ho,

NGUYEN-Duc HOANG PHAM, KIMINORI SHIMIZU,

TOSHIMITSU FUKIHARU, BINH NGUYEN TRUONG, & AKIRA SUZUKI

Xylaria hongkongensis sp. nov. from an urban tree

ALVIN MING CHAK TANG, REGENT YAU CHING LAM,

& MIKE WING KEUNG LEUNG

A new species of Sphaceloma on Helicia from China

HAI-YAN YANG, YUN-YUE WANG, & ZHONG-YI ZHANGT

Additions to the Chilean phalloid mycota PABLO SANDOVAL-LEIVA,

Jose Luis HENRIQUEZ, & LARISSA TRIERVEILER-PEREIRA

New records of cercosporoid fungi from Poland

URSZULA SWIDERSKA-BUREK & WIESLAW MULENKO

New records of Phragmidium species from Pakistan

N.S. AFSHAN & A.N. KHALID

Life cycle of Aecidium araliae and its new name

MAKOTO KAKISHIMA & Qt WANG

Two rare myxomycete species from the Iberian Peninsula

OscCAR REQUEJO & NICANOR FLORO ANDRES RODRIGUEZ

Four lecideoid lichens new to China Line Hu, X1n ZHAO, L1-YAN Sun,

ZUN-TIAN ZHAO, & Lu-LU ZHANG

MARCH-JUNE 2014... V

Muscodor strobeli, a new endophytic species from South India

VINEET MESHRAM, SANJAI SAXENA, & NEHA KAPOOR 93

New record of Setomelanomma holmii on Picea crassifolia in China

based on morphological and molecular data Z1-QIANG Wu,

XIN-LEI FAN, TING YANG, CHENG-MING TIAN,

YInG-MEI LIANG, YAN-FANG Ma, & SAN-LIN ZHANG 105

Thalloloma ochroleucum (Graphidaceae), a new species from

Guizhou, China ZE-FENG JIA & KLAUS KALB 113

A new species of Marasmius sect. Siccifrom India Arun Kumar Dutta,

SWARNENDU CHANDRA, PRAKASH PRADHAN, & KRISHNENDU ACHARYA 117

A new species of Arachnophora from submerged wood in the Amazon

rainforest, Brazil JOSIANE SANTANA MONTEIRO,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL F. CASTANEDA-RUIZ 127

Xiuguozhangia, a new genus of microfungi to accommodate

five described Piricaudiopsis species

Kal ZHANG, LI-Guo Ma, JIAN Ma, & RAFAEL F. CASTANEDA-RUIZ 131

Hymenochaete in China. 8. H. biformisetosa sp. nov. with a key

to species with denticulate setae Jiao YANG & SHUANG-Hut1 HE 137

New records of Dothideomycetes from Mexico

SANTIAGO CHACON, FIDEL TAPIA, & MARTIN ESQUEDA 145

Chaetospermum malipoense sp. nov. from southwest China

X1A0-MING TAN, CHUN-LAN WANG, JUAN CHEN, & SHUN-XING GUO 159

Chemistry and morphology of Chrysothrix candelaris in Poland,

with notes on the taxonomy of C. xanthina

SANDRA OLSZEWSKA, ADRIAN ZWOLICKI, & MARTIN KUKWA 165

Two new species of Diacheopsis from China SHU-ZHEN YAN,

MING-QUAN GUO, & SHUANG-LIN CHEN 173

Australohydnum dregeanum new to Italy

ALESSANDRO SAITTA, MARIA LETIZIA GARGANO,

RICCARDO COMPAGNO, & GIUSEPPE VENTURELLA 179

Anamorphic fungi of the Atlantic Forest of southern Bahia: new records

and Dactylaria pseudomanifesta sp. nov. | DitzE Marta ARGOLO MAGALHAES,

EpDNA DorA MarTINS NEWMAN Luz, ALBERTI FERREIRA MAGALHAES,

Marcos VINICIUS OLIVEIRA Dos SANTOS, FLAVIA RODRIGUES BARBOSA,

LARISSA ARGOLO MAGALHAES, & JOSE LuIz BEZERRA 185

A new slender species of Aureoboletus from southern China

MING ZHANG, TAI-Hut LI, & BIN SONG 195

Nomenclatural revision of four Peziza species

GIANFRANCO MEDARDI, ANGELA LANTIERI, & GABRIELE CACIALLI 203

VI... MYCOTAXON 128

REGIONAL MYCOBIOTA NEW TO THE MYCOTAXON WEBSITE

A checklist of coprophilous fungi and other fungi recorded on dung

from Brazil FRANCISCO JUNIOR SIMOES CALAGA,

NATHAN CARVALHO DA SILVA, & SOLANGE XAVIER-SANTOS 205

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN VOLUME 128 207

ERRATA FROM PREVIOUS VOLUMES

VOLUME 127

Validation of the name Diplococcium variegatum

SILVANA SANTOS DA SILVA, ALISSON CARDOSO RODRIGUES DA CRUZ,

Luis FERNANDO PASCHOLATI GUSMAO, & RAFAEL E CASTANEDA-RUIZ

The proposed name “Diplococcium variegatum S.S. Silva, Gusmao & R.F. Castafieda, sp.

nov.’ on page 60 of volume 127 was not validly published because the cited registration

number, “MB 805030”, was incorrect and is assigned to a separate name. Art. 42.1 of the

International Code of Nomenclature (McNeill & al. 2012) requires that after January 1,

2013, ‘the citation in the protologue of the identifier issued by a recognized repository

for the name (Art. 42.3) is an additional requirement for valid publication’ We hereby

validate the name Diplococcium variegatum S.S. Silva, Gusmao & R.E Castaneda, sp.

nov. with the assigned number MB 805072, with its description published in Mycotaxon

127: 60. 2014 and its holotype designated as HUEFS 194254. Additional details were

published in the original publication.

Other Errata for MyCOTAXON 127:

p. ii, between lines 11 & 12 App: KAREN HANSEN (2014-2019)

Stockholm, Sweden

p.115, line 3 FOR: MILAGRO GRANADOS!

READ: MILAGRO GRANADOS-MONTERO!

MARCH-JUNE 2014...

REVIEWERS — VOLUME ONE HUNDRED TWENTY-EIGHT

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.

N.S. Afshan

Andre Aptroot

Alan W. Archer

Uwe Braun

Peter Buchanan

Gabriele Cacialli

Cecilia Cristina Carmaran

M. Luisa Castro

Manmohan Chhibber

Giovanni Consiglio

Cvetomir M. Denchev

Dennis E. Desjardin

G.S. Dhingra

Matteo Gelardi

Shouyu Guo

Luis EP. Gusmao

José R. Hernandez

Margarita Hernandez-Restrepo

Cheng-Lin Hou

Jae-Seoun Hur

Carlos Antonio Inacio

Alfredo Justo

Roland Kirschner

T.K. Arun Kumar

Carlos Lado

James C. Lendemer

De-Wei Li

Miguel Armando Lopez Ramirez

Cristiano Losi

José G. Marmolejo

Eric H.C. McKenzie

Ireneia Melo

Armin Me&Sic

Jurga Motiejunaite

Karen K Nakasone

A.R. Niazi

Lorelei L. Norvell

Eduardo R. Nouhra

Yoshitaka Ono

Beatriz Ortiz-Santana

Omar Paino Perdomo

Shaun R. Pennycook

Michael J. Richardson

Amy Y. Rossman

Andrea Irene Romero

A.F. Dos Santos

Hyeon-Dong Shin

Sanjay K. Singh

Steven L. Stephenson

Gary A. Strobel

Guangyu Sun

Larissa Vasilyeva

Nadja Santos Vitoria

Robert L. Wick

Y.-J. Yao

Hai-Sheng Yuan

G.I. Zervakis

Tao Zhang

Xiu-Guo Zhang

Wen- Ying Zhuang

PUBLICATION DATE FOR VOLUME ONE HUNDRED TWENTY-SEVEN

MYCOTAXON for JANUARY-MARCH, VOLUME 127 (I-x + 1-234)

was issued on May 29, 2014

VII

vu ... MYCOTAXON 128

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Mycotaxon publishes four volumes a year. Both open access and subscription

articles are offered.

MARCH-JUNE 2014... IX

FROM THE EDITOR-IN-CHIEF

KAREN HANSEN JOINS THE MyYCOTAXON ADVISORY — Through an

unfortunate oversight, we did not list our new Editorial Advisory member in her

rightful place on p. ii of MycoTaxon 127. Karen Hansen (Stockholm, Sweden)

was elected in January to serve as advisor to the Editors during 2014-2019.

We are lucky to have her and take this official opportunity to forward an

appreciative welcome while apologizing profusely for our editorial negligence!

IMC ACCEPTS THREE NOMENCLATURAL REPOSITORIES: On 8 August 2014,

the Tenth International Mycological Congress resolved during its closing

ceremonies in Bangkok to accept “the decision of its Nomenclature Session

with respect to Art. 41 of the INTERNATIONAL CODE OF NOMENCLATURE FOR

ALGAE, FUNGI, AND PLANTS, made 7 August 2014 regarding official repositories

for the registration of fungal names, namely to recognize multiple repositories:

FUNGAL Names, INDEX FUNGORUM, and MYCoBANK ....”

CORRECT NUMBERS REQUIRED FOR VALID PUBLICATION OF NEW NAMES

— The first erratum on p. vi of this volume, slightly more elaborate than usual,

points to the need for authors to scrutinize carefully their nomenclatural

registration numbers before submitting a paper for publication. Because such

numbers are hidden from external inspection until after publication, we — as

editors — can not check them at our end. Remember that the difference of

one digit will refer to a name that differs from what is intended and thereby

invalidates the name. Each number is unique and must be rendered accurately

or the name cannot be regarded as valid.

MyYCOTAXON 128 contains 28 papers by 94 authors (representing 14

countries) and revised by 59 expert reviewers. Within its pages are two new

genera (Anacraspedodidymum —including a new species from Brazil— and

Xiuguozhangia— honoring China's Xiu-Guo Zhang) and 16 other species new

to science representing Arachnophora, Atelocauda, Dactylaria, and Ravenelia

from Brazil; Aureoboletus, Chaetospermum, Diacheopsis, Hymenochaete,

Sphaceloma, Thalloloma, and Xylaria from China; Marasmius and Muscodor

from India; and Tylopilus from Pakistan. In addition to range extensions and/or

new hosts for previously named taxa, five new names (in Peziza and Puccinia)

and 6 new combinations (in Anacraspedodidymum and Xiuguozhangia) are

proposed.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

12 August 2014

MYCOTAXON

http://dx.doi.org/10.5248/128.1

Volume 128, pp. 1-10 April-June 2014

Tylopilus: a new species and a new record from Pakistan

S. SARWAR”*, A.N. KHALID, & A.R. NIAZI

Department of Botany, University of the Punjab, Lahore, 54590, Pakistan

* CORRESPONDENCE TO: samina_boletus@yahoo.com

ABSTRACT — Tylopilus sultanii sp. nov. and T. pseudoscaber are reported from Pakistan,

occurring in forests dominated by ectomycorrhizal trees of Pinaceae. The collections are

described and illustrated, and their habitat is described. Their phylogenetic relationships

were accessed by ITS sequence analysis. Tylopilus pseudoscaber represents a new record for

Pakistan. A key to the three Tylopilus species recorded in Pakistan is presented.

Key worps — Basidiomycota, biogeography, Boletaceae, conifers, Himalaya, phylogeny

Introduction

Khyber Pakhtunkhwa (KPK), Pakistan, which lies at an altitude of 1000-

4000 m in the Himalayan range, is mainly dominated by conifer forests and

regarded as one of the world’s 35 biodiversity hotspots (Myers et al. 2000,

Zachos & Christian 2011). The Mansehra, Dir, Swat, Malakand, and Abbottabad

districts of KPK, and the Rawalpindi district of the Punjab are covered mainly

by conifer forests (Anwar 2008), including fir (Abies), deodar (Cedrus deodara),

Kail (Pinus wallichiana), and chir pine (Pinus roxburghii). Normal growth and

survival of pinaceous trees depends on colonization by ectomycorrhizal (ECM)

fungi (Smith & Read 1997).

The ectomycorrhizal genus Tylopilus P. Karst. has a rich biogeographical

history (Wolfe & Bougher 1993, Halling 1996) and is characterized by smooth

spores, a pinkish to black pore surface, solid stipe lacking an annulus, and

glandular dots (Bessette et al. 2000). Seventy-six Tylopilus species are known

worldwide (Kirk et al. 2008, Osmundson & Halling 2010), but molecular

analysis shows that the genus is polyphyletic (Binder & Hibbett 2006).

Only one Tylopilus species, T. felleus (Bull.) P. Karst., has been reported from

Pakistan (Razaq 2007). We describe here a new species, T: sultanii, and a new

record for the country, T: pseudoscaber. A key to the three Tylopilus species in

Pakistan is also provided.

2 ... Sarwar, Khalid, & Niazi

72° 76°

36°

bia HO AIK

wa i “Bbranel gal

“ @Murree

AFGHANISTAN eae Soe

International Boundaries

District Boundaries

Sampling Sites

ARABIAN

SEA 68°

1 cm = 375 Kilometers

FiGurE 1. Map of Pakistan showing sampling sites.

Materials & methods

Sporocarp collection

Specimens were collected from sampling sites (Fic. 1) in early to late summer (July-

September) during 2006, 2009, and 2011. Field notes and digital photographs were

made from fresh specimens. Sporocarps were dried with hot air from a fan heater and

then kept in paper bags for processing in the lab. Voucher specimens were deposited in

the bolete collection of the Lahore Herbarium, Department of Botany, University of the

Punjab, Lahore, Pakistan (LAH).

Morphological characterization

Characters recorded from fresh sporocarps included:

Piteus: Color, shape, diameter, texture and ornamentation, context color and color

changes, margin shape and color.

HYMENOPHORE: young and mature tube and pore color, tube and pore size, and bruising

reactions of the pore surface.

STIPE: Color, shape, diameter, texture and ornamentation, context color and color

changes, presence/absence of annulus, attachment to the pileus.

Tylopilus sultanii sp. nov. (Pakistan) ... 3

Characters recorded after mounting dried sporocarp tissue in KOH, Meltzer’s

reagent, Trypan blue, and Lactic acid included:

Shape, length, width and cytoplasmic contents of basidia, cystidia, basidiospores,

pileipellis and terminal cells of pileipellis; color reaction of all these in KOH, Meltzer’s,

lactic acid and Trypan blue.

Molecular analysis

DNA was extracted from dried sporocarps by using enzymatic digestion and

glass-fibre filtration (EDGF) protocol (Dentinger et al. 2010). The nuclear ribosomal

internal transcribed spacer region (ITS) was amplified following the PCR conditions of

Dentinger et al. (2010) using primers ITS3 (White et al. 1990) and ITS6R (Dentinger

et al. 2010). PCR products were purified using ExoSAP-IT° (Affymetrix, High

Wycombe, UK) and dye-terminated unidirectional sequencing was performed using a

BigDye*Terminator v3.1Cycle Sequencing Kit (Life Technologies/ABI, California, USA)

in 10uL reactions with primers ITS3 and ITS6R following the protocol of Dentinger

et al. (2010). Sequencing reactions were cleaned using ethanol precipitation following

the manufacturer's instructions, resuspended in 30 uL of distilled water, and run on

an ABI 3730 DNA sequencer in the Jodrell Laboratory, Royal Botanic Gardens Kew.

Sequence chromatograms were edited by comparing overlapping reads using BioEdit

and comparisons to GenBank records using BLAST. Sequences have been deposited in

a single file in GenBank (Tylopilus pseudoscaber: KJ775785; T. sultanii: KJ775786). The

sequences were aligned using MUSCLE alignment software (Edgar 2004). Phylogenetic

trees were constructed with the maximum likelihood algorithm and Jukes & Cantor

(1969) model of sequences evolution using Model-testing feature of MEGA5 software

(Tamura et al. 2011). A bootstrap consensus tree (Fic. 5) was inferred from 1000

replicates, and corresponding bootstrap values >50% are cited in the tree.

Taxonomy

Tylopilus sultanii S. Sarwar, Khalid & Niazi, sp. nov. FIGS 2-3

MycoBank MB 802339

Differs from Tylopilus pseudoscaber by its context that does not change color when

bruised, its larger basidiospores, and its cracked pileus surface.

Type: Pakistan, Khyber Pakhtunkhwa, Ayubia, 2350 m a.s.l., under Pinus wallichiana

A.B. Jacks., on ground, 15 August 2006, A.R. Niazi 36 (Holotype, LAH0806; GenBank

KJ775786).

Erymo_oey: referring to Dr. Sultan Ahmad, a pioneer mycologist in Pakistan.

PILEus 6-9 cm wide, globose, hemispherical to broadly convex, dark brown,

surface dry, rough, cracked into rectangular patches, context pinkish yellow

to creamy visible between patches, margins straight to deflexed, same color as

pileus surface, even margins. CONTEXT creamy, no color change upon exposure.

Odor and taste not distinctive. Stipe about 5 cm long, 3 cm thick, central, solid,

hard, smooth, slightly curved, the center of stipe dark brown to black, at top

and base whitish to creamy in color, context like pileus, no color change upon

A ... Sarwar, Khalid, & Niazi

ss set

oeehee gare

eee het a

‘ TLS uae

Le cameee BS spe

FiGuRE 2. Tylopilus sultanii (holotype, LAH0806).

a, b. Sporocarps, in situ. c,d. Sporocarps. Scale bars = 1.5 cm.

exposure. Pore surface whitish brown to dark brown, adnate and horizontal to

sinuate, pores 2-3 per mm, tubes whitish to off-white, 9-17 mm deep, no color

change to slightly brown upon bruising. Edibility not known.

Basipiospores oblong-inequilateral, apiculate, smooth, 18-19 x 8-10 um,

(18.4 + 0.45 x 9.13 + 0.74; Q. = 2.07 + 0.18). Basrp1A clavate, 2—4-sterigmate,

sterigmata long, contents visible, brownish pigments visible, 62-64 x 19-20

um. CysTrpiA cylindrical to subclavate, thin walled, hyaline, 32-35 x 9-10 um.

PILEIPELLIS a layer of long cylindrical septate hyphae, 76-80(-97) x 10-12 um.

Tylopilus sultanii sp. nov. (Pakistan) ... 5

C mao ds o_-'[|[:

Fic. 3. Tylopilus sultanii (holotype, LAH0806).

a. Basidia. b. Cystidia. c. Pileipellis hyphae. d. Basidiospores.

Scale bars: a = 16 um; b = 9.5 um; c = 17 um; d= 6 um.

CHEMICAL REACTIONS: pileipellis staining reddish brown in KOH; spores

yellowish brown to reddish brown in Meltzer’s reagent.

ECOLOGY & DISTRIBUTION: Known only from the type locality, fruiting July-September

during rainy season.

Notes: Tylopilus sultanii is similar to T: badiceps (Peck) A.H. Sm. & Thiers,

T. indecisus (Peck) Murrill, and T. pseudoscaber. However, T. badiceps differs by

its obliquely truncate pileus, stipe with a whitish apex, reticulations near the

apex, and spores without an apiculus; T’ indecisus differs by its pale brown to

dull cinnamon pileus surface and reticulated stipe; and T: pseudoscaber differs

by its context bluing upon bruising and its smaller basidiospores (Thiers 1975,

Bessette et al. 2000, Lakhanpal 1996).

6 ... Sarwar, Khalid, & Niazi

Tylopilus pseudoscaber (Singer) A.H. Sm. & Thiers, Mycologia 60: 950 (1968) Fra. 4

= Porphyrellus pseudoscaber Singer, Farlowia 2: 115 (1945)

PiLEus 5-9 cm wide, convex to broadly convex, surface dry, smooth, dull,

dark olive brown to dark brown, margins smooth, entire, same color as pileus,

deflexed to slightly incurved. CoNTEXT white to off-white, slowly bluing upon

exposure, and then turning brownish. Stipe 6-11 cm long, 1.5-3 cm thick,

subclavate to equal, straight to slightly curved, dry, dull, solid, centric, color like

pileus surface, whitish near base, sometimes brownish dots present, context

off-white, change to brownish when exposed. Pore surface dark reddish brown

to blackish brown, staining dark blue at first then turning dark brown, pores

2-3/mm, circular to angular, narrow, 2-3 per mm, tubes 1-2.5 cm deep, shorter

towards margins, brownish. Smell pungent, Taste not distinctive. Edibility not

known.

BASIDIOSPORES ellipsoid to subfusoid, smooth, thick walled, 11-16 x 6-8

um, (13.6 + 1.6 x 7.07 0.72; Q_ = 2.05 + 0.37). Basip1a clavate, 3-4 sterigmate,

thick walled, 34-41 x 11-13 um. Cystip1a subclavate to fusoid to ventricose,

with tapering tips, brownish in KOH, 39-59 x 11-17 um. PILEIPELLIs cylindrical

interwoven hyphae, 52-58 x 5-8 um; most terminal elements cylindrical with

pointed tips in some cases, 43-50 x 7-9 um.

CHEMICAL REACTIONS: pileipellis staining yellowish red in KOH, Meltzer’s

reagent; spores ochraceous in Meltzer’s reagent, light yellow in lactic acid.

Eco.oey: In Pakistan, fruiting under coniferous trees, July-September

during the rainy season.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, Khaira gali, 2347

m a.s.l., solitary on ground under Abies pindrow Royle, 17 July 2009, Sarwar S.B. 20

(LAH0709: GenBank KJ775785); Nathiagali, 2439 m a.s.l., solitary on ground under

Pinus wallichiana A.B. Jacks., 19 July 2009, Sarwar S.B. 20A (LAH0709); Sharan (Kaghan

valley), 2011 ma.s.l., solitary on ground under Pinus wallichiana A.B. Jacks., 7 August

2011, Tayiba A. 68 (LAH0811).

Notes: Tylopilus pseudoscaber is similar to T. porphyrosporus (Fr.) A.H. Sm.

& Thiers, T: indecisus, and T: sordidus (Frost) A.H. Sm. & Thiers. However,

T. porphyrosporus differs by its larger size and its context not bluing upon

bruising; T: indecisus differs by its pale brown to dull cinnamon pileus surface

and reticulated stipe; and T: sordidus differs by its longitudinal streaks on the

stipe (Bessette et al. 2000, Thiers 1975).

Key to Tylopilus species recorded from Pakistan

1. Stipe with brownish base, with reticulations .................... 00. eee T. felleus

1. Stipe with whitish base, without reticulations ............ 0... cece eee ee eee eee 2

2. Basidiospores 11-16 x 6-8 um; context bluing upon bruising ...... T. pseudoscaber

2. Basidiospores 18-19 x 8-10 um; context not bluing upon bruising....... T. sultanii

Tylopilus sultanii sp. nov. (Pakistan) ... 7

Figure 4. Tylopilus pseudoscaber (LAH0709).

a, b. Sporocarps. c. Basidia. d. Cystidia. e. Terminal elements of pileipellis hyphae.

f. Pileipellis hyphae. g. Basidiospores.

Scale bars: a, b = 3.5 cm; c = 8 um; d, e = 13 um; f= 14 um; g = 7 um.

Phylogenetic analysis

The closest BLAST matches for our two ITS sequences were T: pseudoscaber

(JF899578) and Porphyrellus pseudoscaber (EU685112); our T. pseudoscaber

(KJ775785) sequence showed 99% similarity with both of these, but our

T. sultanii (KJ775786) sequence showed only <95% similarity.

The maximum likelihood phylogenetic tree (Fic. 5) had 674 genetic

characters after alignment and trimming from both3 and 5 sites of rDNA-ITS.

No characters were excluded from the final analysis, all characters were of the

‘unord’ type, all gaps were treated as “missing” data, and multistate characters

were interpreted as uncertain. The phylogram contained three clades; both of

our sequences clustered in clade I.

8 ... Sarwar, Khalid, & Niazi

Tylopilus pseudoscaber JF899578

26 ly T pseudoscaber KJ775785

Porphyrellus pseudoscaber EU685112 | Sub Clade 1

WT sultanii KJ775786

Tylopilus sordidus EU819450

Tylopilus virens AB509721

100

Clade I

95 Tylopilus virens DQ407256

Tylopilus formosus HM060320

Tylopilus exiguus JN168776

Tylopilus appalachiensis FJ596794

Tylopilus castaneiceps AB289669

Porphyrellus niger JF908792

Tylopilus felleus JF908787

Tylopilus vinosobrunneus AB509871

Tylopilus atronicotianus EU685114 eeakin

Tylopilus chromapes AB509881

66 Tylopilus leucomycelinus JF908789

98 Tylopilus ballouii AB509735

Tylopilus felleus JN182869

Tylopilus plumbeoviolaceoides DQ407261 ‘| Clade III

Tylopilus rubrobrunneus GQ166869

Boletus edulis EU231984 _]Out group

0.02

FiGurE 5. Phylogenetic position of Tylopilus sultanii and T. pseudoscaber from Pakistan relative to

other Tylopilus spp., with Boletus edulis as outgroup. Tree inferred by maximum likelihood analysis

based on 5.8S+ITS2 rDNA sequences. Numbers against branches indicate percentage support

(>50%) in 1000 bootstrap replications. GenBank accession numbers stand after species names.

= = new sequences from Pakistan.

Tylopilus sultanii was shown to be phylogenetically closest to Porphyrellus

pseudoscaber (EU685112), with which it shares 91.2% genetic characters while

being separated by a 1.9% genetic divergence. There was significant genetic

divergence of rDNA-ITS between T° sultanii and all other rDNA sequences

included in the present study. Maximum similarity and shared genetic

characters were well below the 97% cutoff value for species delimitation, and

genetic divergences were high enough to confirm this species as new.

Our T. pseudoscaber sequence shared 99.9% genetic characters with both

T. pseudoscaber (JF899578) and Porphyrellus pseudoscaber (EU685112) with

only 0.2% genetic divergence.

Discussion

Both morphological and molecular analysis confirm T!: sultanii as a new

species, forming a highly supported sister clade to T: pseudoscaber.

Tylopilus sultanii sp. nov. (Pakistan) ... 9

Our sequence analysis confirmed our morphological identification of our

Pakistani T. pseudoscaber collections, which formed a very highly supported

clade with North American sequences of T. pseudoscaber. This is a new record

for Pakistan.

Acknowledgments

We are thankful to reviewers, Prof. Dr. Robert Laurence Wick (University of

Massachusetts, Amherst, USA) and Dr. Najam-ul-Sehar Afshan (University of the Punjab,

Lahore, Pakistan). We are also very grateful to the Higher Education Commission of

Pakistan for providing financial support for this research work and Jodrell Laboratories

(Royal Botanic Gardens Kew) for providing facilities for molecular work.

Literature cited

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Verlag Dr. Muller Aktiengesellschaft and Co. KG, Germany.

Bessette A, Roody WC, Bessette AR. 2000. North American boletes: a color guide to the fleshy

pored mushrooms. Syracuse Univ Pr.

Binder M, Hibbett DS. 2006. Molecular systematics and biological diversification of Boletales.

Mycologia 98: 971-981. http://dx.doi.org/10.3852/mycologia.98.6.971

Dentinger BTM, Margaritescu S, Moncalvo JM. 2010. Rapid and reliable high-throughput methods

of DNA extraction for use in barcoding and molecular systematic of mushrooms. Molecular

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Edgar RC. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput.

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Halling RE. 1996. Boletaceae (Agaricales): latitudinal biodiversity and biological interactions in

Costa Rica and Colombia. Revista de Biologia Tropical 44: 111-114.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th ed. Wallingford:

CABI.

Lakhanpal TN. 1996. Mushrooms of Indian Boletaceae. Vol. I, in: KG Mukherji (ed.). Studies in

cryptogamic botany. APH Publishing Corporation, Delhi.

Myers N, Mittermeier RA, Mittermeier CG, Fonseca GAB, Kent J. 2000. Biodiversity hotspots for

conservation priorities. Nature 403: 853-858. http://dx.doi.org/10.1038/35002501

Osmundson TW, Halling RE. 2010. Tylopilus oradivensis sp. nov.: a newly described

member of the Tylopilus balloui complex from Costa Rica. Mycotaxon 113: 475-483.

http://dx.doi.org/10.5248/113.475

Razaq A. 2007. Taxonomic studies on Basidiomycota from Northern Areas of Pakistan. PhD thesis,

University of Karachi, Karachi.

Smith SE, Read DJ. 1997. Mycorrhizal symbiosis. 2‘ Edition. Academic Press London. 605 p.

Tamura K, Peterson P, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGAS: molecular

evolutionary genetics analysis using maximum likelihood, evolutionary distance, and

maximum parsimony methods. Molecular Biology and Evolution.

http://dx.doi.org/10.1093/molbev/msr121

Thiers HD. 1975. California mushrooms. A field guide to the boletes. New York, NY: Hafner Press.

Wallenberg foundation, Sweden.

10 ... Sarwar, Khalid, & Niazi

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

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

methods and applications. Academic Press, San Diego.

Wolfe CB, Bougher NL. 1993. Systematics, mycogeography, and evolutionary history of Tylopilus

subg. Roseoscabra in Australia elucidated by comparison with Asian and American species.

Australian Systematic Botany 6: 187-213.

Zachos FE, Christian HJ. 2011. Biodiversity hotspots: distribution and protection of conservation

priority areas. Heidelberg; New York: Springer.

MY COTAXON

http://dx.doi.org/10.5248/128.11

Volume 128, pp. 11-15 April-June 2014

Anacraspedodidymum, a new genus

from submerged wood in Brazil

CAROLINA RIBEIRO SILVA’, RAFAEL FE. CASTANEDA-RUIZ’,

& Luis FERNANDO PASCHOLATI GUSMAO?*

"Departamento de Micologia, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil

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

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

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

*Departamento de Ciéncias Biologicas, Laboratério de Micologia, Universidade Estadual

de Feira de Santana, BR116 KMO3, 44031-460, Feira de Santana, Brazil

* CORRESPONDENCE TO: lgusmao@uefs.br

ABSTRACT — Anacraspedodidymum aquaticum gen. et sp. nov., found on submerged

decaying wood in Riacho do Mel, Bahia, Brazil, is described and illustrated. It is characterized

by monophialidic, integrated, terminal conidiogenous cells, with a funnel shaped collarette

and solitary, obovoid, globose to elliptical, slightly apiculate at the base, aseptate, hyaline

conidia with irregular to subreticulate mucous ornamentation. A new combination,

Anacraspedodidymum hyalosporum, is proposed to accommodate Craspedodidymum

hyalosporum.

KEY worps — microfungi, freshwater, taxonomy

Introduction

During research on conidial fungi associated with submerged decaying

plant material in Riacho do Mel, municipality of Alagoinhas, Bahia, Brazil, an

interesting fungus was found, distinguished by phialidic conidiogenous cells and

conidial features that show some similarities with the genus Craspedodidymum

Hol.-Jech. but which morphologically differs from all described species.

Therefore, it is described as new to science.

Samples of submerged litter were placed in paper and plastic bags. In the

laboratory the samples were placed in Petri dish moist chambers and stored ina

170 L polystyrene box with 200 mL sterile water plus 2 mL glycerol, at 25°C for 30

days (Castafieda-Ruiz 2005). Mounts were prepared in PVL (polyvinyl alcohol,

lactic acid, and phenol) and measurements were made at a magnification of

12 ... Silva, Castafieda, & Gusmao

x1000. Micrographs were obtained with an Olympus microscope (model

BX51) equipped with bright field and Nomarski interference optics. The type

specimens are deposited in the Herbarium of Universidade Estadual de Feira

de Santana (HUEFS).

Taxonomy

Anacraspedodidymum C.R. Silva, R.F. Castafieda & Gusmao, gen. nov.

MycoBank 805402

Differs from Craspedodidymum by its hyaline basal apiculate conidia, sometimes with

mucilaginous adherences.

TYPE SPECIES: Anacraspedodidymum aquaticum C.R. Silva et al.

EryMo_oey: Greek, Ana-, meaning upwards, back and again; Latin, -craspedodidymum

referring to the hyphomycete genus Craspedodidymum.

Co.onliss effuse, hairy, brown to black. Mycelium superficial and immersed.

CONIDIOPHORES macronematous, mononematous, unbranched, straight

to slightly flexuous, septate, smooth or verruculose, brown to pale brown.

CONIDIOGENOUS CELLS monophialidic, integrated, terminal, cylindrical,

sometimes with percurrent elongations; collarette funnel shaped or

infundibuliform, brown. Conrp1a solitary, aseptate, solitary, ellipsoid, spherical,

globose, obpyriform to obovoid, with an inconspicuous basal hilum or slightly

papillate, sometimes with mucous adherences or tunicate ornamentation,

hyaline.

Anacraspedodidymum aquaticum C.R. Silva, R.F. Castafieda & Gusmao,

sp. nov. PLATE 1

MycoBAnk 805403

Differs from Craspedodidymum spp. by its hyaline conidia with irregular or subreticulate

mucilaginous ornamentations.

Type: Brazil, Bahia State, Alagoinhas, Riacho do Mel, 12°10’S 38°24’W, on submerged

decaying wood in a stream, 18 Oct. 2011, coll. C.R. Silva (Holotype: HUEFS 196431).

ErymMo.oey: Latin, aquaticum, refers to its growing in water.

CoLonlEs effuse, hairy, brown. Mycelium partly superficial, partly immersed

in the substratum. Hyphae septate, smooth, pale brown, 2.5 um diam.

CONIDIOPHORES macronematous, mononematous, simple, unbranched,

straight to slightly flexuous, 2-7-septate, smooth, brown to pale brown at

apex, 64-219 x 3-5 um. CONIDIOGENOUS CELLS monophialidic, integrated,

terminal, cylindrical, sometimes with percurrent elongations, 17-63 x 3-4 um;

collarette funnel shaped, 4-5 x 2-3 um, pale brown. Conip1a solitary, ellipsoid,

spherical to obovoid, aseptate, with irregular or subreticulate mucilaginous

ornamentation, hyaline, 8-12 x 7-8 um, mostly slightly papillate at the base.

Anacraspedodidymum gen. nov. (Brazil) ... 13

Fic. 1. Anacraspedodidymum aquaticum (holotype, HUEFS 196431). A. Conidiophores. B. Conidia.

C. Conidiogenous cells and conidia. Scale bars: A = 20 um; B-C = 5 um.

14 ... Silva, Castafeda, & Gusmao

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, Banta, Riacho do Mel, 12°10’S 38°24’W,

on submerged wood, 18 Oct. 2011, coll. C.R. Silva (HUEFS 196430).

Notes: Holubova-Jechova (1972) and Seifert et al. (2011) described

Craspedodidymum (type = C. elatum Hol.-Jech.) with dichotomously branched

or single conidiophores and phialidic terminal conidiogenous cells with a

funnel-shaped collarette; the conidia are solitary, 0-4-septate, brown to dark

brown. The Craspedodidymum generic concept was enlarged to accommodate

C. hyalosporum, a species with 1-septate, hyaline conidia (Bhat & Kendrick,

1993) and which differs from all other described Craspedodidymum species

(Yanna et al. 2000, Ma et al. 2011). We propose a new combination in

Anacraspedodidymum.

Anacraspedodidymum hyalosporum (Bhat & W.B. Kendr.) R.E. Castafieda,

C.R. Silva & Gusmao, comb. nov.

MycoBank 805404

= Craspedodidymum hyalosporum Bhat & W.B. Kendr., Mycotaxon 49: 35 (1993).

Bahusutrabeeja Subram. & Bhat, Cylindrotrichum Bonord., Kylindria

DiCosmo et al., and Monilochaetes Halst. are also superficially similar to

Anacraspedodidymum, but Bahusutrabeeja has aseptate hyaline conidia with

several filiform extracellular appendages. Cylindrotrichum has polyphialidic

conidiogenous cells, while monophialidic conidiogenous cells are present

in Kylindria, but blastic sympodial extension frequently occurs within the

collarette in both genera (Castafieda-Ruiz & Kendrick 1990, Kirk et al. 2008,

Seifert et al. 2011). Monilochaetes has monophialidic conidiogenous cells, with

an inconspicuous or evident collarette and cylindrical to oblong conidia that

accumulate in mucous masses or are (rarely) pseudocatenulate (Reblova et al.

2011, Seifert et al. 2011)

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li and Dr. M. Hernandez-

Restrepo for their critical review of the manuscript. The authors thank the Programa de

Pés-Graduacao em Biologia de Fungos(PPGBF/ UFPE). C.R. Silva and LFP Gusmao

extend their gratitude to CNPq for financial support (Proc. 132415/2013-5 and

305413/2001-2). RFCR is grateful to Cuban Ministry of Agriculture and “Programa

de Salud Animal y Vegetal’, project P131LH003033 from Cuban Ministry of Science,

Technology and Environment for facilities. We acknowledge the facilities provided by

Dr. P.M. Kirk and Drs. V. Robert and G. Stegehuis through the Index Fungorum and

Mycobank websites. Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s

nomenclature review are greatly appreciated.

Literature cited

Bhat DJ, Kendrick WB. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Anacraspedodidymum gen. nov. (Brazil) ... 15

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

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

Castafeda Ruiz RE, Kendrick WB. 1990. Conidial fungi from Cuba II. University of Waterloo

Biology Series. 33: 1-61.

Holubova-Jechova V. 1972. Craspedodidymum, a new genus of phialosporous hyphomycetes Ceska

Mykologie 26: 70-73.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi. 10th ed. CAB

International, UK, Wallingford.

Ma LG, Ma J, Zhang YD, Zhang XG. 2011. Craspedodidymum and Corynespora spp.

nov. and a new anamorph recorded from southern China. Mycotaxon 117: 351-358.

http://dx.doi.org/10.5248/117.351

Reblova M, Gams W, Seifert KA. 2011. Monilochaetes and allied genera of the Glomerellales,

and a reconsideration of families in the Microascales. Studies in Mycology 68: 163-191.

http://dx.doi.org/10.3114/sim.2011.68.07

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

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

Yanna, Ho WH, Goh TK, Hyde KD. 2000. Craspedodidymum nigroseptatum sp. nov., a new

hyphomycete on palms from Brunei Darussalam. Mycol. Res. 104: 1146-1151.

MY COTAXON

http://dx.doi.org/10.5248/128.17

Volume 128, pp. 17-23 April-June 2014

Two new rust species on Fabaceae from Brazil

MAuvRICIO SALAZAR YEPES! & ANIBAL ALVES DE CARVALHO JUNIOR”

"Universidad Nacional de Colombia, sede Medellin, Calle 59A, No. 63-20, Medellin, Colombia

?Instituto de Pesquisas Jardim Botdnico do Rio de Janeiro,

Pacheco Ledo 915, CEP 22460-030, Rio de Janeiro, RJ, Brazil

* CORRESPONDENCE TO: anibal@jbrj.gov.br

ABSTRACT— Two new species of rust fungi (Basidiomycota, Pucciniales) on Fabaceae are

proposed: Atelocauda sakuraguiae on Ormosia sp. and Ravenelia costae on Pseudopiptadenia

leptostachya. Both were collected at the Parque Nacional do Itatiaia, Brazil. Atelocauda

sakuraguiae can be distinguished from all other Atelocauda species on Fabaceae by its non-

lobed aeciospores, septate aecial paraphyses, less elongate teliospores, and septate teliospore

pedicels. Ravenelia costae can be distinguished from all other Ravenelia species by its smooth

teliospores, probasidia deriving from germ pores, pendent cysts, echinulate urediniospores,

and absence of paraphyses.

Key Worps— Atlantic forest, biodiversity, phytopathology, plant disease, Uredinales

Introduction

Through a project supported by the Instituto de Pesquisas Jardim Botanico

do Rio de Janeiro entitled “Uredobiota (fungi) do Parque Nacional do Itatiaia,”

rust fungi specimens were collected in the States of Minas Gerais and Rio

de Janeiro, Brazil. This field and laboratory work revealed two species with

distinctive characteristics differing from other rust species on Fabaceae that are

described here as new.

Materials & methods

The specimens were collected during 2006 and 2007 in the Parque Nacional do

Itatiaia in the States of Minas Gerais and Rio de Janeiro, Brazil. Collections were slowly

dried and accessioned into the Herbarium of the Instituto de Pesquisas Jardim Botanico

do Rio de Janeiro (RB). Sori were observed using a Leica S6E stereomicroscope. Free

hand sections and scrapings were placed in lactophenol or chloral hydrate and then

examined using a Zeiss Axioskop 40 compound microscope. For scanning electron

microscopy (SEM) observation, material was mounted on a double-sided adhesive tape,

fixed in osmium vapor for 17 h, gold-palladium coated with an Emitech K550X coater,

18 ... Salazar Yepes & Carvalho Junior

and observed with a Zeiss EV 550X SEM. Illustrations were created from images of

these slides.

Taxonomy

Atelocauda sakuraguiae Salazar-Yepes & A.A. Carvalho, sp. nov. Fics 1-8

MycosBaNnk MB564578

Differs from all other Atelocauda species on Fabaceae by its non-lobed aeciospores,

septate aecial paraphyses, less elongate teliospores, and septate teliospore pedicels.

TyPE: on Ormosia sp. (Fabaceae): Brazil, Minas Gerais, Parque Nacional do Itatiaia, Vale

de Santa Clara Trail, 22°1917.4”S 44°37'17.8’W, 1615 m asl, 11 Oct 2007, M. Salazar

Yepes & A.A. Carvalho Jr. 638-07 (Holotype, RB480639).

EryMoLoecy: Dedicated to Cassia Monica Sakuragui, a distinguished botanist and

authority on Araceae.

Spermogonia few, amphigenous, mostly on adaxial leaf surface, Group VI

(type 7). Aecia Uredo-type, few amphigenous, mostly on adaxial leaf surface,

subcuticular, in groups, erumpent, cinnamon-brown. Paraphyses abundant,

peripheral, incurved, flexuous, septate at base, 81-90 x 9-12 um; wall smooth,

colorless to pale yellow. Aeciospores obovoid to clavate, 27-30 x 21-27 um; wall

echinulate, 1.5-2 um thick at sides, 2-3 um thick at base, chestnut gold; germ

pores obscure, one near the hilum. Telia on abaxial leaf surface, scattered, or in

groups, minute, subcuticular in origin, becoming ruptured with conspicuous

ruptured cuticle, pulverulent, chocolate-brown, with abundant peripheral

paraphyses. Paraphyses cylindrical, flexuous, incurved, dichotomically and

trichotomically branched, septate at base, 75-105 x 12-15 um; wall smooth,

mostly dorsally thickened up to 6 um, and colorless to pale yellow. Teliospores

obovoid to oblong-globoid, 24-33 x 24-30 um; distal tubercles, 5 um wide;

wall uniformly 2-3 um thick, cinnamon brown, tuberculate; germ pores one

near the hilum; pedicel fragile, septate near the spore, <75 um long, colorless.

EcoLocy — The rust infects Ormosia plants at altitudes above 1600 m

above sea level (asl) in National Park of the Itatiaia, whose forest is designated

as dense rainforest High Montana (Veloso et al. 1991).

CoMMENTs — Arthur & Cummins (1933) erected Atelocauda (with

A. incrustans Arthur & Cummins as its type) differing from Uromyces by its type

7 spermogonia and from Dicheirinia and Diabole by its one-celled teliospores

formed on pedicels. Thirumalachar & Kern (1955) reduced the genus to

synonymy under Pileolaria and recombined its type as P. incrustans. Although

Cummins (1959, 1978) accepted the placement in Pileolaria, Cummins &

Hiratsuka (1983) resurrected Atelocauda, to which they transferred three

Uromyces species on Acacia from Hawaii, Australia, and neighboring regions.

Walker (2001), who re-circumscribed Atelocauda, transferred five species

assigned to Atelocauda by Cummins & Hiratsuka (1983), Ono (1984), and

Atelocauda & Ravenelia spp. nov. (Brazil) ... 19

on adaxial leaf surface. 2. Section of spermogonium (type 7). 3. Flexuous paraphyses showing

a septum (arrowed). 4. Aeciospores; left, median focus; right, surface focus. 5. Section of telial

sorus, showing teliospores and paraphyses. 6. Dichotomous telial paraphyses. 7. Teliospores;

left, median focus; right, surface focus. 8. Teliospores; pedicel septate near the spore (arrowed).

Bars: 2, 5 = 50 um; 3, 4, 7, 8 = 20 um; 6 = 10 um.

Gardner (1991) to a new genus Racospermyces and added a new species,

A. shivasii J. Walker on Ormosia from Australia. Zhuang & Wei (2009)

recombined Uromyces ormosiae L. Guo & Y.C. Wang on Ormosia spp. as

Atelocauda ormosiae, based on its subcuticular spermogonia type 7.

Atelocauda shivasii, A. incrustans, and A. ormosiae differ from A. sakuraguiae

by their lobed anamorph spores, their non-septate teliospore pedicels, and their

more elongated teliospores (Arthur & Cummins 1933, Walker 2001, Zhuang &

Wei 2009).

20 ... Salazar Yepes & Carvalho Junior

Walker (2001) also discovered spermogonia and Uredo-type aecia with lobed

spores on the type specimen of A. incrustans and pointed out the morphological

similarities between Atelocauda shivasii on Ormosia and Dicheirinia ormosiae

(Arthur) Cummins, whose urediniospores are similar in shape with a basal

germ pore associated with a smooth patch. Similarly Atelocauda sakuraguiae

has Uredo-type aecia and aeciospores with a basal germ pore in a smooth area

(Fic. 4). From these morphological similarities, it is assumed that Atelocauda

is more closely related to Raveneliaceae than to Pileolariaceae (sensu Cummins

& Hiratsuka 2003). Like Walker (2001), we consider Atelocauda to be closely

related to Ravenelia, Dicheirinia, and other genera in Raveneliaceae.

Ravenelia costae Salazar-Yepes & A.A. Carvalho, sp. nov. Fics 9-14

MycosBank MB564579

Differs from all other Ravenelia species by its smooth teliospores, probasidial cells

germinating by germ pores, pendent cysts, echinulate urediniospores, and absence of

paraphyses.

Type: on Pseudopiptadenia leptostachya (Benth.) Rauschert (Fabaceae): Brazil, Rio de

Janeiro, Itatiaia, Parque Nacional do Itatiaia, around the bridge over the Rio Campo

Belo, 22°27'41.1”S 44°37'9.5”W, 1109 m asl, 6 Aug 2007, M. Salazar Yepes & A.A.

Carvalho Jr. 469-07 (Holotype, RB532381).

ErymMoLoecy: Dedicated to Denise Pinheiro da Costa, a distinguished botanist and

authority on Metzgeriaceae, Hepaticae.

Spermogonia amphigenous, in groups on rounded necrotic lesions, 1-2 mm

diam. Aecia dark cinnamon brown, on abaxial leaf surface, scattered, or in

groups, peridiate, cupulate, white to pale yellow. Peridial cells irregular in

shape, firmly united, 21-48 um diam.; outer wall finely verrucose and inner

wall verrucose-striate, colorless. Aeciospores obovoid to oblong-ellipsoid,

polyhedric, 30-42 x 24-27 um; wall verrucose, 1-2 um thick, pale chestnut

brown to colorless. Uredinia on abaxial leaf surface, scattered, or in small

groups, minute, subepidermal in origin, ruptured epidermis conspicuous,

pulverulent, cinnamon-brown; paraphyses absent. Urediniospores obovoid,

pyriform, ellipsoid, 30-45 x 18-24 um; wall echinulate, 1-2 um thick at

sides, 1-3 um thick at apex, pale cinnamon-brown; germ pores obscure.

Telia on abaxial leaf surface, scattered, or in small groups, subepidermal in

origin, ruptured epidermis conspicuous, pulverulent, dark cinnamon brown;

paraphyses absent. Teliospore heads 45-90 um diam., 4-7 probasidial cells

across, central cells 18-21 um diam., variable in size and shape; wall smooth,

2-3 um thick, cinnamon-brown; cysts colorless, globoid, pendent, 21-30 um

diam.; germ pores evident, 1 in each cell, 3-6 um diam.; pedicel multi-hyphal,

mostly persistent, <75 um, hyaline.

ADDITIONAL SPECIMENS EXAMINED: on Pseudopiptadenia leptostachya: BRAZIL,

RIO DE JANEIRO, Itatiaia, Parque Nacional do Itatiaia, on the Véu de Noiva-Itaporani

Atelocauda & Ravenelia spp. nov. (Brazil) ... 21

FIGURE 9-14. Ravenelia costae (holotype, RB532381). 9. Peridial cells. 10. Aeciospores; left,

median focus; right, surface focus. 11. Urediniospores. Left, median focus; right surface focus.

12. Teliospores; upper view (SEM). 13. Teliospores; left, median focus; right, surface focus.

14. Teliospores; apical pores (arrowed). Bars = 20 um.

Waterfalls Trail, 1180 m asl, 17 Aug 2006, M. Salazar Yepes, A.A. Carvalho Jr. & I. Franca

227-06 (RB480597); 23 Apr 2007, M. Salazar Yepes, A.A. Carvalho Jr. & E Santoro 211-07

(RB480593); Rio Campo Belo-Prateleiras Trail, 22°25’44.8’S 44°37’37.6’W, 1400 m

asl, 5 Dec 2006, M. Salazar Yepes & A.A. Carvalho Jr. 448-06 (RB480621); 22°26’7.3”S

44°37'32.5”W, 1265 m asl, 27 Apr 2007, M. Salazar Yepes, A.A. Carvalho Jr. & F. Santoro

355-07 (RB480612); around the Abrigo 10 (Lamedo), 22°25’34.4”S 44°37'54.1” W, 1500

m asl, 5 Dec 2007, M. Salazar Yepes & A.A. Carvalho Jr. 725-07 (RB480644).

22 ... Salazar Yepes & Carvalho Junior

EcoLocy — The rust-infected P. leptostachya plants are found at altitudes

under 1500 m in National Park of the Itatiaia, whose forest is classified as dense

rainforest Montana (Veloso et al. 1991).

CoMMENTS — Pseudopiptadenia, the host of our species, belongs to the

Piptadenia group in the Mimosoideae subfamily of legumes. Mimosoideae

hosts have at least 73 species of Ravenelia. From these, only seven have smooth

teliospores and uredinia that (if present) lack paraphyses. Ravenelia versatilis

(Peck) Dietel and R. chacoensis J.C. Lindq. differ from R. costae by their larger

teliospores; R. bahiensis Henn., R. hieronymi Speg., and R. entadae Lagerh. &

Dietel differ by their teliospores having germ slits, and R. bahiensis also differs

by being microcyclic; R. striatispora Cummins & J.W. Baxter differs by its

urediniospores having striate ornamentation; and R. prosopidicola J.C. Lindq.

differs by its smaller aeciospores with 6-7 equatorial germ pores (Hennen et al.

2005, Resende 2007, and Hernandez et al. 2012).

After comparison with the most comprehensive recent literature, including

Hennen et al. (2005), Resende (2007), and Hernandez et al. (2012), we

determined that R. costae is unique among other Ravenelia by the combination

of smooth teliospores, probasidial cells germinating by germ pores, pendent

cysts, echinulate urediniospores, and uredinia without paraphyses.

Acknowledgments

We express our deep appreciation to Prof. Dr. Yoshitaka Ono and Prof. Dr. José

R. Hernandez for their valuable suggestions in reviewing this manuscript. We thank

the Fundac¢ao Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro

(FAPERJ - Rio de Janeiro, Brazil) and Conselho Nacional de Pesquisa Cientifica e

Tecnoldgica (CNPq - Brazil). In addition, our institutions, including the Universidad

Nacional de Colombia, sede Medellin, and the Instituto de Pesquisas Jardim Botanico

do Rio de Janeiro (JBRJ), have made it possible to continue this research.

Literature cited

Arthur JC, Cummins GB. 1933. New species of Uredinales. Ann. mycol. 31: 41-45.

Cummins GB. 1959. Illustrated genera of rust fungi. Minneapolis, MN: Burgess Publishing Co.

131 p.

Cummins GB. 1978. Rust fungi on legumes and composites in North America. Tucson, AZ:

University of Arizona Press. 424 p.

Cummins GB, Hiratsuka Y. 1983. Illustrated genera of rust fungi. Revised edition. St Paul: American

Phytopathological Society. 152 p.

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi. Third edition. St Paul: American

Phytopathological Society. 225 p.

Gardner DE. 1991. Atelocauda angustiphylloda n.sp., a microcyclic rust on Acacia koa in Hawaii.

Mycologia 83: 650-653. http://dx.doi.org/10.2307/3760220

Hennen JF, Figueiredo MB, Carvalho Junior AA, Hennen PG. 2005. Catalogue of the species of

plant rust fungi (Uredinales) of Brazil. 490 p. http://www.jbrj.gov.br (accessed: 19 Jun 2012).

Atelocauda & Ravenelia spp. nov. (Brazil) ... 23

Hernandez JR, Hennen JE, Farr DE, McCray EB. 2012. Ravenelia online, Systematic Mycology and

Microbiology Laboratory, ARS, USDA.

http://nt.ars-grin.gov/taxadescriptions/keys/ravenelialndex.cfm (accessed: 19 Jun 2012).

Ono Y. 1984. A monograph of Maravalia (Uredinales). Mycologia 76: 892-911.

http://dx.doi.org/10.2307/3793146

Resende DV. 2007. Sistematica de Uredinales da familia Raveneliaceae e espécies do Cerrado

brasileiro. Revisio Anual de Patologia de Plantas (RAPP) 15: 99-172.

Thirumalachar MJ, Kern FD. 1955. The rust genera Allotelium, Atelocauda, Coinostelium and

Monosporidium. Bull. Torrey Bot. Club 82(2): 102-107. http://dx.doi.org/10.2307/2481825

Veloso HP, Rangel Filho ALR, Lima JCA. 1991 . Classificagao da vegetacao brasileira, adaptada a

um sistema universal. Rio de Janeiro: IBGE, Departamento de Recursos Naturais e Estudos

Ambientais. 124 p.

Walker J. 2001. A revision of the genus Atelocauda (Uredinales) and description of Racospermyces

gen. nov. for some rusts of Acacia. Austral. Mycol. 20: 3-28.

Zhuang J-Y, Wei S-X. 2009. Notes on some rust fungi from southern China. Mycosystema 28:

630-636.

MY COTAXON

http://dx.doi.org/10.5248/128.25

Volume 128, pp. 25-36 April-June 2014

The first record of Hebeloma vinosophyllum (Strophariaceae)

in Southeast Asia

Bao-THUY QUYEN Ho”, NGUYEN-Duc HOANG PHAM3,

KIMINORI SHIMIZU‘, TOSHIMITSU FUKIHARU?,

BINH NGUYEN TRUONG®, & AKIRA SUZUKI”?

" Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo 271-8510, Japan

? Faculty of Biology, University of Science, Vietnam National University in Hochiminh City,

227 NguyenVanCu St., Dist. 5, Hochiminh City, Vietnam

> Division of Microbiology, Biotechnology Center of Hochiminh City,

km 1900, Highway 1A, Dist. 12, Hochiminh City, Vietnam

* Medical Mycology Research Center, Chiba University,

1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan

° Natural History Museum and Institute, Chiba,

955-2 Aoba-cho, Chuo-ku, Chiba 260-8682, Japan

° Faculty of Biology, Dalat University,

1 Phu Dong Thien Vuong St., Dalat City, LamDong Province, Vietnam

’ Faculty of Knowledge Engineering, Tokyo City University,

1-28-1 Tamazutsumi, Setagaya-ku, Tokyo 158-8557, Japan

* CORRESPONDENCE TO: thquyen21@yahoo.com

ABSTRACT - Basidiomata of a Hebeloma sp. were collected from two urea-plots in a pine

(Pinus kesiya) forest, Da Lat City, Lam Dong Province, Vietnam. Based on morphology,

molecular phylogeny, and mating compatibility, the specimens were identified as Hebeloma

vinosophyllum. This is the first record of H. vinosophyllum in Southeast Asia and of

H. vinosophyllum occurring in P. kesiya forests, one of two dominant pines in Southeast Asia.

Key worps - Porphyrospora, cortina, cheilocystidia, ammonia fungi

Introduction

Hebeloma vinosophyllum was originally described by Hongo (1965) from

Japanese specimens. This species was reported to resemble H. sarcophyllum

(Peck) Sacc. in having a light brown to purplish red spore print but to differ in

the shape of the hymenial cystidia (Hongo 1965, Singer 1986, Vesterholt 2005).

In 1975, Sagara reported H. vinosophyllum as a member of the “ammonia fungi

26 ... Ho &al.

... a chemo-ecological group of fungi that develop reproductive structures

exclusively or relatively luxuriantly on soil after a sudden addition of ammonia,

of other nitrogenous materials that react as bases by themselves or on

decomposition, or of alkalis” (Sagara 1975). Basidiomata of H. vinosophyllum

were recorded not only from the forest floor sites augmented with urea but also

from soils naturally disturbed by decomposition of dead animal bodies (Sagara

1976, 1992, Takayama & Sagara 1981, Fukiharu et al. 2000a,b).

Hebeloma vinosophyllum has been recorded from different vegetation in

various geographical regions of Japan. The fungus has been collected from

Castanopsis, Quercus, and Pinus forests (TABLE 1) in middle and western

Honshu (Hongo 1965, Sagara 1975, Suzuki 1987, Yamanaka 1995, Fukiharu et

al. 1995, Fukiharu et al. 2000a,b, Kasuya 2002, Imamura & Yumoto 2004), and

Shikoku and Kyushu (Sagara 1975). However, this fungus has not been recorded

for urea applications in Pinus and Quercus forests in Hokkaido (Sagara 1975) or

eastern Honshu (Sagara 1975, Fukiharu & Horigome 1996), or in Castanopsis

and Quercus forests on Ryukyu Island (Fukiharu & Hongo 1995).

Outside Japan, H. vinosophyllum has been recorded from a fagaceous-

coniferous forest (Quercus, Castanopsis, Castanea, Cunninghamia, Pinus, etc.)

near SanMing City, Fujian Province, China (Hongo et al. 1996; no voucher

specimen deposited - Dr. Noriko Kinjo, pers. comm.) and possibly in South

Korea (Lee 2011; species name listed without details). These collection records

suggest that H. vinosophyllum has an East Asian distribution (Fukiharu &

Horigome 1996).

During a survey of ammonia fungi in southern Vietnam, specimens

belonging to Hebeloma sect. Porphyrospora Konrad & Maubl. ex Vesterh.

(Vesterholt 2005) were collected. We identified these Vietnamese specimens

based on morphology, molecular phylogeny, and compatibility between the

strains from Vietnam and stock cultures of H. vinosophyllum from Japan.

Materials & methods

Collection

Urea-treated plots were established in a Pinus kesiya Royle ex Gordon forest (more

than 30 years old), ca. 1500 m altitude, at Xuan Tho, Da Lat City, Lam Dong Province,

Vietnam. Commercial granulated urea fertilizer was applied to 2 plots (1 x 1 m) on

the forest floor in the amount of 600 g/m’. About two months after urea application,

basidiomata of Hebeloma sp. were observed, collected, and subcultured onto agar

medium. Cultures were maintained on MY agar medium [malt extract 10 g/L (Difco,

Detroit, USA), yeast extract 2 g/L (Difco, Detroit, USA) and agar 15 g/L (Nacalai Tesque,

Japan)] at 20°C in darkness. Basidiomata were dried at 60°C for 24 h and deposited

in the herbarium of the Natural History Museum and Institute, Chiba, Japan (CBM).

Cultures were deposited in Faculty of Biology, University of Science, Ho Chi Minh City,

Vietnam (HCMUS).

Hebeloma vinosophyllum in Vietnam ... 27

Other specimens and cultures of H. vinosophyllum in Japan, H. aminophilum

in Australia, and H. porphyrosporum in Italy were deposited in the Natural History

Museum and Institute, Chiba, Japan (CBM); Forest Microbiology Division, Forestry

and Forest Products Research Institute, Ibaraki, Japan (CHU); and Biological Resource

Center, National Institute of Technology and Evaluation, Japan (NBRC).

Observation

Morphological observations were recorded from both dry and fresh field specimens.

Color notation follows Kornerup & Wanscher (1981).

The microscopic characteristics were observed using differential interference

contrast (DIC) on Labophot-2 (Nikon, Tokyo, Japan) or Olympus B51 (Tokyo, Japan)

microscopes after hand-sectioning and mounting in 10% ammonium hydroxide

aqueous solution. The basidiospores were also observed by a scanning electron

microscope (SEM) (Hitachi S-800; Tokyo, Japan) at 15.0 kV after rehydrating in 25%

aqueous ammonia, fixing in 2.5% osmic acid, and coating with platinum-palladium in

an ion sputter-coater (E-1030; Hitachi, Tokyo, Japan). Abbreviations used: Q = mean

length/width ratio measured from “n” number of spores; m = mean spore length and

width.

Phylogenetic analysis

Total nuclear DNA (nDNA) was extracted from disintegrated tissue of dried mycelia

and basidiomata using 200 ul 0.5 mm glass beads (Yasui Kikai, Tokyo, Japan) and 500 ul

TES buffer [50 mM Tris-HCl, pH 7.5, 20 mM EDTA, 1% sodium dodecyl sulfate (SDS)]

by vigorous shaking (FastPrep System; mp-Biomedicals, Solon, OH, USA) at 6.5 m/s for

45 s. Soluble fractions were recovered by centrifugation. DNA was purified using TE

buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA)-saturated phenol/chloroform/isoamy1

alcohol (25:24:1, Nippon Gene, Tokyo, Japan) extraction followed by an isopropyl alcohol

precipitation. After desiccation of the DNA pellet, DNA was dissolved in a 30 ul TE

buffer. For some samples, genomic DNA was further purified using NucleoSpin Extract

II (Macherey-Nagel, Duren, Germany), following the manufacturer's recommendations.

The primer pairs ITS1 and ITS4 or ITS5 and ITS4 (White et al. 1990) were used to

amplify the ITS regions of ribosomal DNA (rDNA). The primers LROR and LRS (Vilgalys

and Hester 1990) were used to amplify 28S rDNA (large subunit, LSU). Polymerase chains

reactions (PCR) were carried out using KOD FX (Toyobo, Tokyo, Japan) following the

manufacturer's instructions. PCR products were purified using NucleoSpin Extract II,

and DNA fragments were directly sequenced using the BigDye Terminator ver3.1 Cycle

Sequencing Kit (Applied Biosystems, Foster City, CA, USA) following the protocol

provided. Reactions were cleaned up using the Centri Sep (Princeton Separations,

Adelphia, NJ, USA), before analyzing by capillary electrophoresis on a 3130x DNA

Analyzer (Applied Biosystems). Sequences were assembled and edited using ATSQ

software (Genetyx, Tokyo, Japan) and deposited in GenBank/EMBL/DDJB (TaBLE 1).

Two data sets (LSU, ITS) were established that included sequences from Vietnamese

specimens, several sequences of Japanese H. vinosophyllum, and sequences downloaded

from GenBank (TABLE 2). The data sets were aligned using Clustal X ver. 1.81

(Jeannmougin et al. 1998), and the resulting alignments were manually refined. For

phylogenetic analyses, each data set (LSU = 892 bp, ITS = 625 bp) was analyzed using

28 ... Ho &al.

TABLE 1. Collection details of Hebeloma specimens and cultures in the phylogenetic

analysis.

GENBANK ACC. NO.

TAXON VOUCHER NO./ISOLATE NO. ORIGIN

ITS LSU

H. vinosophyllum CBM:FB32636*/CBM-BC69 Japan AB1742172 ABI1742341

CBM:FB12306*/CBM-BC307 Japan AB1742173 AB1742342

CBM:FB12325*/CBM-BC314 Japan AB1742174 AB1742343

CBM:FB12381*/CBM-BC315 Japan AB1742175 AB1742344

CBM:FB14335*/CBM-BC337 Japan AB1742176 AB1742345

CBM:FB14285*/CBM-BC366 Japan AB1742177 AB1742346

CBM:FB14216*/CBM-BC376 Japan AB1742178 AB1742347

CBM:FB14520*/CBM-BC384 Japan AB1742179 AB1742348

CBM:FB14502*/CBM-BC407 Japan AB1742180 AB1742349

CBM:FB15552*/CBM-BC437-3 Japan AB1742181 AB1742350

CBM:FB15553*/CBM-BC438-2 Japan AB1742182 AB1742351

CBM:FB15556*/CBM-BC440-2 Japan AB1742183 AB1742352

CBM:FB24700/CBM-BC533** Japan AB1742184 AB1742353

—/NBRC107913 Japan AB1742185 AB1742354

—/CHU4001* Japan AB1742186 AB1742355

CBM:FB39191*/HCMUS-Cl1 VietNam AB1742187 AB1742356

CBM:FB39192*/— VietNam AB1742188 AB1742357

CBM:FB39267*/HCMUS-C2 VietNam AB1742189 AB1742358

H. aminophilum —/CHU5001* Australia AB1742190 AB1742359

—/CHU5002*** Australia ABI1742191 AB1742360

CBM:FB35472*/— Australia AB1742192 AB1742361

H. porphyrosporum — CBM:FB24804/— Italy AB1742193 AB1742362

Basidiomata appeared * in urea plot, ** around decaying unidentified animal, or *** around decaying snake.

Tree-Puzzle 5.2 (Schmidt et al. 2002). The maximum likelihood (ML) tree (Felsenstein

1981) of each data set was inferred based on quartet puzzling algorithm (Strimmer &

Haeseler 1996) with the options of 1000 puzzling steps, model of substitution HKY

(Hasegawa et al. 1985).

TABLE 2. Published sequences in the phylogenetic analysis

GENBANK ACC. NO.

wt ITS LSU

Hebeloma fastibile AF325643 AY033139

H. sarcophyllum AF124715 ——

H. vinosophyllum

03294501]* ——

(NBRC32945)

H. vinosophyllum AY320398** ——

(Boyle et al. 2006)

Alnicola lactariolens AY818352 AY818353

Anamika angustilamellata AY575919 AY575919

An. indica AF407163 AF407164

* Retrieved from DNA resource of Biological Resource Center,

National Institute of Technology and Evaluation, Japan.

** Used only for alignment with sequence from Vietnamese H. vinosophyllum.

Hebeloma vinosophyllum in Vietnam ... 29

Mating test

The dikaryotic stock culture of Vietnamese Hebeloma sp. (HCMUS-C2) was

cultured on Ohta’s medium (Ohta 1990) with 15 g agar, at 20°C, 12 h/12 h light/dark

cycle. Light was irradiated at ca. 140 umol m”’s"'. Basidiomata appeared in vitro after ca.

3 weeks, when a small piece of sterile filter paper was placed under the hymenophores

for collecting basidiospores. Monokaryotic strains were obtained by germinating these

in vitro spores.

Mating tests between monokaryotic tester strains of Vietnamese Hebeloma sp.

and dikaryotic stock cultures of Japanese H. vinosophyllum (isolates CBM-BC69

and CHU4001) were compared. Mating types were determined by pair-mating with

each other. The mating tests were conducted by placing a pair of mycelial discs (a

monokaryotic tester strain vs. a dikaryotic culture) on opposite sides of 90 mm MY

agar plates, incubated at 25°C in darkness, and replicated 3 times. After two weeks,

mycelium from the monokaryotic colony was removed from the edge farthest from the

dikaryotic colony and examined microscopically; a compatible crossing was indicated

by the presence of clamp connections.

"> GO ED

A B C D

PLaTE 1. Hebeloma vinosophyllum. A-D: Hymenial cystidia. E: Basidiospores.

Scale bar = 10 um.

Taxonomy

Hebeloma vinosophyllum Hongo, J. Jap. Bot. 40: 314. 1965 PLaTEs 1-2

[Description based on Vietnamese specimens] Pileus 20-70 mm broad,

at first hemispherical, then becoming plano-convex to plane; cream to light

30 ... Ho & al.

PLATE 2. Hebeloma vinosophyllum from the urea plot in Viet Nam. A: Basidiomata in nature.

B: Cortina (partial veil; *) of a young basidioma. C: Scanning electron micrographs of basidiospores.

D: Ixotrichodermium structure of pileipellis. Scale bars: C = 3 um; D = 30 um.

vinaceous (7-9A2-3), usually darker at the top; surface glabrescent, smooth,

viscid to glutinous in wet condition; margin at first inrolled and entire, then

plane or eroded-undulating, sometimes still incurved at maturity. Universal veil

absent, partial veil fibrillose, white, dry, at first cortinoid (following Vesterholt

2005), leaving a few fibrils on the stipe, disappearing with age. Lamellae

crowded, 2-4 mm width, 5-30 mm length, adnexed; pinkish brown (8-9C3-5)

to light vinaceous (8-9A3-5) and darker at maturity. Stipe up to 105 x 15

mm, thickened downward; surface somewhat smooth to longitudinal striate,

floccose-pruinose at the apex; fleshy-fibrous, stuffed at first, then hollow; cream

(7A2-3) to light clay (5D5-6). Context pale white (7-8A2-3); smell musty;

taste weakly bitter. Spore print vinaceous to brownish red (9-10C3-4).

Basidiospores m = 9.7 + 0.6 x 6.1 £ 0.4 um (8.4-11.3 x 5.3-7.2 um, n = 50,

Q = 1.59), amygdaliform to citriform. Basidia 30-40 x 8-9 um (without

sterigmata), clavate, four-spored; sterigmata 3-5 um long. Pleurocystidia

48-60 x 8-12 um, numerous, ventricose-rostrate to lageniform, narrowly

utriform or mucronate. Cheilocystidia similar to pleurocystidia but less

numerous. Pileipellis an ixotrichodermium, composed of gelatinized cylindrical

cells 50-60 um in length; hypodermium cellular composed of globose cells 7-9

um in width.

Hebeloma vinosophyllum in Vietnam ... 31

Hasirat - In Vietnam, scattered, gregarious, or subcespitose on urea-plots

in Pinus kesiya forest, ca. 1500 m, 2 months after urea application. Elsewhere in

warm temperate East Asia scattered, gregarious, or subcespitose on soil under

Castanopsis, Quercus, and Pinus, usually 6-12 months after urea application or

following decomposition of dead animal bodies.

DISTRIBUTION (PLATE 3): Japan (middle to western Honshu, Shikoku, and

Kyushu); China (SanMing, Fujian Province); Vietnam (DaLat City, LamDong

Province); possibly South Korea (confirmation needed).

SPECIMENS EXAMINED: VIETNAM. Lam Downe: Da Lat Crry, Xuan Truong - Xuan

Tho (UTM, 49P, 023053, 1319015), 24 Jun 2009, B.N Truong (CBM FB-39191; culture

HCMUS-C1); 25 Jun 2009, B.N Truong (CBM FB-39192); 14 Jun 2009, B.N Truong

(CBM FB-39267; culture HCMUS-C2). JAPAN. SHIGA: Otsu City, Ishizune, 17 Jul

1961, T Hongo (TNS-F-39101, holotype)

PLATE 3. Biogeographic distribution of Hebeloma vinosophyllum in Japan (Honshu: Saitama,

Tochigi, Tokyo, Chiba, Shiga, Shizuoka, Kyoto, Tottori; Shikoku: Kochi; Kyushu: Oita, Miyazaki),

South Korea, China (Fujian Province), and Vietnam (Da Lat). A = no deposition of voucher

specimen, O = collection site of voucher specimen.

32 ... Ho &al.

98 An. indica AF407163

An. angustilamellata AY575919

Al. lactariolens AY818352

H. aminophilum CBM:FB35472

H. aminophilum CHU5001

H.aminophilum CHU5002

H. fastibile AF325643

100 H. sarcophyllum AF124715

L H. porphyrosporum CBM:FB24804

H. vinosophyllum CBM:FB39192

H. vinosophyllum NBRC107913

H. vinosophyllum CBM:FB39191

H. vinosophyllum CBM:FB39267

H. vinosophyllum CBM-BC314

H. vinosophyllum CBM-BC366

H. vinosophyllum CBM-BC533

H. vinosophyllum CBM-BC315

H. vinosophyllum NBRC32945

H. vinosophyllum CBM-BC440-2

H. vinosophyllum CBM-BC376

H. vinosophyllum CBM-BC307

H. vinosophyllum CBM-BC384

H. vinosophyllum CBM-BC69

H. vinosophyllum CBM-BC407

H. vinosophyllum CHU4001

H. vinosophyllum CBM-BC437-3

H. vinosophyllum CBM-BC438-2

H. vinosophyllum CBM-BC337

0.01

An. indica AF407164

An. angustilamellata AY575919

ner H. porphyrosporum CBM:FB24804

H. fastibile AY033139

Al. lactariolens AY818353

99 H. aminophilum CBM:FB35472

H. aminophilum CHU5002

H. aminophilum CHU5001

. vinosophyllum NBRC107913

. Vinosophyllum CBM:FB39192

. vinosophyllum CBM-BC69

, vinosophyllum CBM-BC366

, vinosophyllum CBM-BC337

. vinosophyllum CBM-BC315

. vinosophyllum CBM-BC407

|, vinosophyllum CBM-BC440-2

|. vinosophyllum CHU4001

. vinosophyllum CBM:FB39267

H. vinosophyllum CBM-BC533

. vinosophyllum CBM-BC438-2

. Vinosophyllum CBM-BC437-3

. Vinosophyllum CBM-BC384

|. vinosophyllum CBM-BC376

. vVinosophyllum CBM-BC314

| vinosophyllum CBM-BC307

. Vinosophyllum CBM:FB39191

XZ} @TTTIITIIIT

mrrIIT

0.01

PLaTE 4. The maximum likelihood phylogenetic tree based on ITS data set (A) and LSU data

set (B) of Hebeloma vinosophyllum and allied species. The numbers on each branch represent

bootstrap percentage support values. The scale bar shows the ratio substitutions/site. Trees are

rooted by outgroup Anamika indica and An. angustilamellata. Vietnamese specimens are in bold.

Abbreviation: Al. = Alnicola.

Phylogenetic analysis

Dikaryotic cultures and dried specimens of Vietnamese H. vinosophyllum

have identical ITS and LSU sequences. Sequence alignments of dikaryotic

H. vinosophyllum cultures from Japan and dried H. vinosophyllum specimens

Hebeloma vinosophyllum in Vietnam ... 33

from Vietnam showed 99.55% (887/891 bp) similarity of the LSU and 98.52%

(600/609 bp) similarity of the ITS regions. The phylogenetic tree (PLATE 4)

grouped the dikaryotic cultures and the Vietnamese specimens in the same

clade as those from Japan (supporting values = 68% in LSU and 86% in ITS).

We could not analyze the holotype specimen (TNS-F-39101) of

H. vinosophyllum molecularly due to its long storage with para-formaldehyde.

The lower ITS similarity of 95.4% (587/615 bp) for GenBank AY320398

(Boyle et al. 2006, labeled as H. vinosophyllum) suggests that that material is

misidentified.

Mating test

The di-mon mating tests between monokaryotic strains from Vietnam and

dikaryotic cultures of Japanese H. vinosophyllum showed clamp connections

(TABLE 3), indicating that all Vietnamese monokaryotic strains were compatible

with Japanese H. vinosophyllum cultures.

TABLE 3. Dikaryon-monokaryon mating tests between Hebeloma vinosophyllum

(Japan) and Hebeloma sp. (Vietnam)*

Monokaryotic strains of Vietnamese Hebeloma sp.

HCMUS-C2

C2-03 C2-36 C2-02 C2-37 C2-09 C2-38 C2-01 C2-35

Dikaryotic stock

cultures of

Japanese

H. vinosophyllum

CBM-BC69 + + + + + + + +

CHU4001 + + + + + + + +

*+ = clamp connections formed

Discussion

Except for forming more abundant cheilocystidia, all Vietnamese specimens

were similar to the holotype specimen of H. vinosophyllum (TNS-F-39101) based

on morphological analysis. The di-mon mating tests indicate that Vietnamese

Hebeloma sp. belongs to the same biological species as H. vinosophyllum. The

phylogenetic analyses (PLATE 4) also indicate that all Vietnamese and Japanese

specimens belong to the same species.

This is the first record of H. vinosophyllum in Southeast Asia (PLATE 3) and

of H. vinosophyllum associating with P. kesiya, one of two dominant pines in

Southeast Asia (Zonneveld et al. 2009).

As a putative ectomycorrhizal fungus (Fukiharu 1991, Sagara 1995,

Imamura & Yumoto 2008), H. vinosophyllum could habituate from hosts of

34 ... Ho &al.

Northeast Asian fagaceous and/or pinaceous species to those of Southeast

Asian Pinus species, or vice versa. Further studies of applications of urea to

forests up to the northern limit of P kesiya in central Asia (Zonneveld et al.

2009) and to fagaceous (possibly Quercus and Castanopsis) and pinaceous

forests in Ryukyu islands, Taiwan, southern China, and Southeast Asia will help

resolve the geographic range, host tree species, and potentially the origin of

H. vinosophyllum.

Despite the presence of a cobweb-like partial veil (cortina), H. vinosophyllum

could belong to Porphyrospora (Vesterholt 2005) based on its brownish red

spore deposit and amygdaliform-citriform spores.

Acknowledgments

This work was financially supported in part by a Grand-in-Aid for the Scientific

Research (Nos. 04660156, 19570082) from the Japan Society for the Promotion of

Science (JSPS). This work was also financially supported in part by Fujiwara Natural

History Foundation, Japan (fiscal year: 1994-1995), Bilateral Exchanging Program

(Australia) (fiscal year 1996). We are thankful to Alcoa World Alumina Australia, and

The University Forest in Chiba - The University of Tokyo for their support and making

the experimental site available for collecting isolates. We wish to thank Prof. Emeritus

Naohiko Sagara (Kyoto University, Japan) for providing the fungal isolates and his useful

information. We would like to give thanks to Dr. Noriko Kinjo (formerly on the Academic

staff of Tokyo Medical and Dental University, Japan) for her important information.

We would like to express sincere thanks to Dr. Inez C. Tommerup (the former senior

researcher of CSIRO Forestry and Forest Products, Wembley, Australia) and Dr. Neale

L. Bougher (Western Australian Herbarium, Australia) for their supporting us to make

an experimental site available for collecting isolates from a Eucalyptus forest. We are

grateful to Prof. Dennis E. Desjardin (San Francisco State University, San Francisco,

USA) and Dr. Peter K. Buchanan (Landcare Research, Auckland, New Zealand) for

reviewing of the manuscript.

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Hebeloma vinosophyllum in Vietnam ... 35

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MY COTAXON

http://dx.doi.org/10.5248/128.37

Volume 128, pp. 37-40 April-June 2014

Xylaria hongkongensis sp. nov. from an urban tree

ALVIN MING CHAK TANG*??, REGENT YAU CHING LAM?”,

& MIKE WING KEUNG LEUNG’

* Muni Arborist Limited, Rm.206B, Sun Cheong Building,

1 Cheung Shun Street, Lai Chi Kok, Kowloon, Hong Kong, China

? School of Life Sciences, The Chinese University of Hong Kong,

Shatin, Hong Kong, China

* CORRESPONDENCE TO: alvin.tang@muniarborist.com

ABSTRACT — A new species of Xylaria from Hong Kong, China, is described based on

its teleomorphic characters and is distinguished by its conical stromata and small spores.

The fungus was found on the trunk of a living Bauhinia variegata in an urban park, and

the infected tree exhibited overall declining symptoms including branch dieback and leaf

discoloration.

KEY worDs — taxonomy, wood decay fungi, Xylariales

Introduction

Trees in cities constitute the most prominent elements of urban nature

due to their seasonal changes, size, shape, and color (Tyrvainen et al. 2005).

However, they are beset by many aerial and subterranean stresses, making

them susceptible to fungi (Bradshaw et al. 1995, Luley 2005, Sinclair & Lyon

2005). Loss of mechanical strength caused by fungal damage in urban areas

can be linked to hazardous situations such as tree wind throws or limb failures,

resulting in significant property damage and injuries. Therefore, timely

assessment and accurate identification of wood decay fungi are important for

assessing structural stability and predicting the probability of tree failure or

decline.

During an assessment of trees on an urban slope, a species of Xylaria was

found on a declining Bauhinia variegata. The tree exhibited dieback symptoms

with leaf discoloration and browning. Numerous stromata emerged on one

dead trunk and along bark cracks of the declining trunk. Morphological

features showed that this fungus represents an undescribed species of Xylaria,

which is described and illustrated here.

38 ... Tang, Lam, & Leung

Materials & methods

Fungal samples were collected on a slope of Waterfall Bay Park, Waterfall Bay Road,

Wah Fu, Southern District, Hong Kong, China. Microscopic observations were made in

water, and Melzer’s reagent was used to test the amyloidity of apical ring. Measurements

were taken at x400 magnification on samples of 30 ascospore, 20 asci, and 10 perithecia.

A herbarium specimen was deposited in the Institute of Microbiology, Chinese Academy

of Science, Guangzhou, China (GDGM), and a living culture placed in Centraalbureau

voor Schimmelcultures, The Netherlands (CBS).

Taxonomy

Xylaria hongkongensis A.M.C. Tang, R.Y.C. Lam & M.W.K. Leung, sp.nov. Fic. 1

MycoBank MB 807096

Differs from Xylaria pallide-ostiolata by its cylindric-clavate stromata with conical apices

and shorter and lighter-colored ascospores.

Type: China, Guangdong Province, Hong Kong, Southern District, Wah Fu, Waterfall

Bay Road, Waterfall Bay Park, on trunk of Bauhinia variegata L. (Caesalpiniaceae),

14.1.2011, A.M.C. Tang (Holotype, GDGM40058; ex-type culture, CBS 136698;

GenBank KF926669).

EryMoLocy — Referring to the type locality region.

STROMATA upright, solitary or caespitose, unbranched, nearly sessile to slightly

stipitate, cylindric-clavate, sterile apices conical to subglobose, 38-67 mm total

length, 8-12 mm diameter; surface dark brown, roughened with wrinkles.

PERITHECIA with slightly exposed outlines, subglobose, 410-840 um high,

410-690 um diam. OsTIOLEs slightly papillate or inconspicuous. Ascr 8-spored,

cylindrical, long-stipitate, 120-147.5 um total length, 3.75-7.5 um width, the

spore-bearing part 67.5-95 um long, with apical apparatus bluing in Melzer’s

reagent, tubular-shaped, 3-4.5 um high x 2-3 um broad. Ascospores brown to

dark brown, unicellular, ellipsoid-inequilateral, 12.5-15 x 5-7.5 um, smooth,

usually with a straight germ-slit nearly spore length on the concave side.

ComMENtTs — ‘This new Xylaria possesses a combination of morphological

characters that do not match any previously described species. Based on

its stromatal appearance, X. hongkongensis should be categorized in the

X. polymorpha complex. Its ascospores resemble those of X. pallide-ostiolata

Henn. (12-18 x 6-7 um; San Martin & Rogers 1989: 356) and Xylaria sp.

SM303 (12.5-14.5 x 5-5.5 um; San Martin & Rogers 1989: 364-365), but

X. pallide-ostiolata differs by its broadly clavate stromata with rounded apices

Fic. 1. Xylaria hongkongensis (holotype, GDGM40058). a—b: Stromata on dead trunk. c: Stromata

emerged on cracks of trunk. d: Herbarium specimen GDGM40058. e: Stromatal surface with

ostioles. f: Section through stroma, showing perithecial layer. g: Ascus in water. h: Apical apparatus

of immature asci stained in Melzer’s reagent. i: Ascospores observed in water. j: Ascospores

observed in water showing germ slit morphology. Scale bars: a—b = 5 cm; c-d = 1 cm; e-f = 500

um; g-j = 10 um.

Xylaria hongkongensis sp. nov. (China) ...

AO ... Tang, Lam, & Leung

and its longer black spores, while Xylaria sp. SM303 differs by a germ-slit that is

much less than spore length and its longer asci (148-166 um). Xylaria scruposa

(Fr.) Fr., which has similarly sized stromata (15-60 x 2-6 mm), differs by its

narrow stroma with a highly roughened surface and longer ascospores (18-22

x 6-7 um; San Martin & Rogers 1989: 363).

It is noteworthy that X. hongkongensis was found in association with trunk

and basal rot in a common urban tree during our tree risk assessment. ‘This

association reminds us that some Xylaria species could manifest themselves

as pathogens on weak hosts, especially in susceptible urban plantations. Since

Xylaria species are ubiquitous endophytes of most vascular plants (Petrini et al.

1995), under favourable conditions Xylaria species might change their passive

ecological strategy into an active mode (Promputtha et al. 2007). Xylaria species

have long been recognized as causing root rot on alfalfa, apple, crabapple,

eastern redbud (Cercis canadensis), hedge maple (Acer campestre), pear, and

thornless honeylocust (Gleditsia triacanthos var. inermis) (Rogers 1984, Proffer

1998). Thus, it is important to stay vigilant on trees showing signs of stress.

Acknowledgments

We wish to express our gratitude to Dr. Eric McKenzie (Landcare Research, New

Zealand) and Dr. Larissa Vasilyeva (Institute of Biology & Soil Science, Far East Branch

of the Russian Academy of Sciences) for reviewing the manuscript. Thanks are also

given to Ms. Hermia Chung and Patty Chu for their technical assistance.

Literature cited

Bradshaw AD, Hunt B, Walmsley T (eds). 1995. Trees in the urban landscape: principles and

practice. Spon, London.

Luley CJ. 2005. Wood decay fungi common to urban living trees in the northeast and central

United States. Urban Forestry LLC, Naples, New York.

Petrini O, Petrini LE, Rodrigues K. 1995. Xylariaceous endophytes: an exercise in biodiversity.

Fitopatol. Bras. 20: 531-539.

Profter TJ. 1998. Xylaria root rot of urban trees caused by Xylaria polymorpha. Plant Disease 72: 79.

http://dx.doi.org/10.1094/PD-72-0079A

Promputtha I, Lumyong S, Dhanasekaran V, McKenzie EHC, Hyde KD, Jeewon R. 2007. A

phylogenetic evaluation of whether endophytes become saprotrophs at host senescence.

Microbial Ecology 53(4): 579-590. http://dx.doi.org/10.1007/s00248-006-9117-x

Rogers JD. 1984. Xylaria acuta, Xylaria cornu-damae, and Xylaria mali in continental United States.

Mycologia 76: 23-33.

San Martin Gonzalez F, Rogers JD. 1989. A preliminary account of Xylaria of Mexico. Mycotaxon

34: 283-373.

Sinclair WA, Lyon HH. 2005. Diseases of trees and shrubs, 2nd edition. Cornell University Press,

Ithaca.

Tyrvainen B, Pauleit S, Seeland K, de Vries S. 2005. Benefits and uses of urban forests and trees.

81-114, in: CC Konijnendijk et al. (eds). Urban forests and trees. Berlin Heidelberg, Springer.

http://dx.doi.org/10.1007/3-540-27684-X_5

MYCOTAXON

http://dx.doi.org/10.5248/128.#

Volume 128, pp. 41-43 April-June 2014

A new species of Sphaceloma on Helicia from China

HAI-YAN YANG?”, YUN-YUE WANG? & ZHONG-YI ZHANG

' Chuxiong Normal University, Chuxiong, Yunnan 675000, China

’ Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China,

Yunnan Agricultural University, Kunming, Yunnan 650201, China

“CORRESPONDENCE TO: wangyykm@gmail.com

ABSTRACT—Sphaceloma heliciae sp. nov. on Helicia formosana is described and illustrated.

‘The type specimen was collected from Xishuangbanna, Yunnan Province, China.

Key worps—coelomycetes, identification, taxonomy

Introduction

The genus Sphaceloma, proposed by de Bary in 1874, contains 163 named

species (Index Fungorum 2013), about 52 of which are currently accepted

(Kirk et al. 2008). Many species have Elsinoe teleomorphic states. In China,

24 Sphaceloma spp. have been reported (Tai 1979; Wang et al. 1999; Guo 2001,

2005). Of the five Sphaceloma spp. that have been recorded on proteaceous

hosts (Elsinoe banksiae Pascoe & Crous, E. leucospermi L. Swart & Crous,

E. proteae Crous & L. Swart, Sphaceloma banksiicola Pascoe & Crous, and

S. protearum L. Swart & Crous), none has been reported on Helicia (Swart et

al. 2001; Pascoe & Crous 2007). We describe a new species found on leaves of

Helicia from China, and distinguish it from the other five species on Proteaceae.

The holotype specimen is conserved in Herbarium Mycologica of Yunnan

Agricultural University, Kunming, China (MHYAU).

Sphaceloma heliciae H.Y. Yang & Z.Y. Zhang, sp. nov. Fic. 1

MycoBAank804587

Differs from the Sphaceloma states of Elsinoe banksiae and E. leucospermi by its shorter,

narrower conidiophores and its narrower, fusiform to ovoid conidia.

Type: China, Yunnan, Xishuangbanna, in living leaves of Helicia formosana Hemsl.

(Proteaceae), Dec. 2009, Li-Xin (Holotype, MHYAU 13083).

EryMo_oey: referring to the host genus.

42 ... Yang, Wang, & Zhangt

00000() °

20pm Sum

Fic. 1. Sphaceloma heliciae (holotype, MHYAU 13083): A. acervulus; B. conidia.

Scabs subrotund, amphigenous, ochroleucous, prominent, 0.8 x 1 mm. Acervuli

subcuticular, scattered or aggregated, 31.0 x 51.7 um, composed of cornuted

cells, hyaline to brown. Conidiophores compactly arranged, continuous,

hyaline, 13.1-23.9 x 1.7-3.5 um. Conidia fusiform, ovoid or occasionally

cylindrical, continuous, hyaline, 4.6-6.2 x 1.3-2.1 um.

Discussion

The hosts and morphology of Sphaceloma heliciae and the five other species

on proteaceous hosts are presented in TABLE 1. In addition to their association

with different host genera, the five other species differ morphologically

from S. heliciae by their broader ellipsoidal conidia and by either their

broader conidiophores (Elsinoe banksiae, E. leucospermi, S. banksiicola, and

S. protearum) or their lack of extended conidiophores (E. proteae).

TABLE 1. Host associations and morphology of Sphaceloma/Elsinoe species

on proteaceous hosts.

CONIDIOPHORES CONIDIA

SPECIES Host

(uum) (uum)

E. banksiae Banksia 20-30 x 3-5 4-6 x 2-3

serrata subcylindrical ellipsoid

E. leucospermi Leucospermum 20-30 x 3-6 5-7 x 2.5-3

cordifolium subcylindrical ellipsoid

E. proteae Protea reduced to hi

, a jnnb

cynaroides conidiogenous cells

S. protearum Protea 12-20 x 5-6 5-6 x 2-2.5

eximia subcylindrical ellipsoid

S. banksiicola Banksia 8-20 x 3-5 8-9 x 2.5-4

prionotes subcylindrical ellipsoid

S. heliciae Helicia 13-23 x 0.7-3.5 4.6-6.2 x 1.3-2.1

formosana subcylindrical fusiform to ovoid

Sphaceloma heliciae sp. nov. (China) ... 43

Acknowledgments

The authors would like to express their sincere thanks to Dr Shaun Pennycook for

reading the manuscript and giving constructive suggestions. Our thanks also go to Prof.

G.Y. Sun and Prof. T. Zhang for reading the manuscript and serving as pre-submission

reviewers. This study was supported by the National Natural Science Foundation of

China (No. 2006 FY120100).

Literature cited

Guo YL. 2001. Imperfect fungi. 131-240, in: WY Zhuang (ed.). Higher fungi of tropical China.

Mycotaxon Ltd., Ithaca NY.

Guo YL. 2005. Anamorphic fungi. 125-232, in: WY Zhuang (ed.). Fungi of Northwestern China.

Mycotaxon Ltd., Ithaca NY.

Index Fungorum. 2013. http://www.indexfungorum.org/names/Names.asp (accessed 22/10/2013).

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi,

10th ed. CABI International, Wallingford, UK. 771 p.

Pascoe I, Crous PW, Groenewald JZ. 2007. Sphaceloma banksiicola Pascoe & Crous, sp. nov. Fungal

Planet no. 14.

Swart L, Crous PW, Kang JC, Mchau GRA, Pascoe I, Palm ME. 2001. Differentiation of species of

Elsinoe associated with scab disease of Proteaceae based on morphology, symptomatology, and

ITS sequence phylogeny. Mycologia. 93(2): 366-379. http://dx.doi.org/10.2307/3761658

Tai FL.1979. Sylloge Fungorum Sinicorum. Beijing: Science Press. 1527 p. (in Chinese)

Wang YZ, Wu SH, Zhou WN, Chang DZ, Chen KY, Chen SF, Chen CL, Tzeng SS, Liou JH, Hsieh

WH, Hsieh HR, Chung CH, Chien CY. 1999. List of the fungi in Taiwan. Committee for

Agriculture, Taipei. 289 p. (in Chinese)

MYCOTAXON

http://dx.doi.org/10.5248/128.45

Volume 128, pp. 45-54 April-June 2014

Additions to the Chilean phalloid mycota

PABLO SANDOVAL-LEIVA’, JOSE LuIs HENRIQUEZ?’,

& LARISSA TRIERVEILER-PEREIRA?>*

'Biota Gestion y Consultorias Ambientales Ltda.,

Miguel Claro 1224, Providencia, Santiago, Chile.

*Departamento de Sanidad Vegetal, Facultad de Ciencias Agronomicas, Universidad de Chile,

Casilla 1004, Santiago, Chile.

°PPGBOT, Universidade Federal do Rio Grande do Sul,

Av. Bento Goncalves 9500, Porto Alegre, Rio Grande do Sul, Brazil

*CORRESPONDENCE TO Lt_pereira@yahoo.com.br

ABSTRACT — Two species in the Phallales (Agaricomycetes), Laternea pusilla and Lysurus

cruciatus, are recorded as new to Chile. We also present a discussion and a key to the phalloid

species known to occur in the country.

Key worps — Clathrus, gasteromycetes, Ileodictyon, Phallus, stinkhorns

Introduction

Phalloid fungi, commonly known as stinkhorns, may produce basidiomata

with very distinct morphologies, but all are characterized by an ovoid structure

during the immature stage. Most species produce an expanded basidiome when

mature, but others (e.g., in Protubera Moller, Claustula K.M. Curtis, Gelopellis

Zeller, maintain the glebal tissue enclosed by the peridium in a sequestrate

structure. The gleba is usually mucilaginous and fetid, which facilitates the

spores’ dispersion by insects.

Phallales E. Fisch. is traditionally classified in two families, mainly based on

basidiome development and morphology: Clathraceae Chevall. and Phallaceae

Corda (Fischer 1898-99, Pilat 1958, Kirk et al. 2008). Recent sequence analyses

support these two families as monophyletic clades, but four additional families

are accepted: Claustulaceae G. Cunn., Lysuraceae Corda, Protophallaceae Zeller,

and Trappeaceae P.M. Kirk (Hosaka et al. 2006, Kirk et al. 2008).

Phalloids are widespread, but most species are found in the tropics, with

South East Asia being the center of diversity (Kreisel 1996, Cannon & Kirk

2007). However, despite their ample diversity and distribution, in Chile they

46 ... Sandoval-Leiva, Henriquez, & Trierveiler-Pereira

have been poorly studied. The first report is from the nineteenth century, when

Montagne (1853-54: 497-499) recorded the occurrence of species of Clathrus

P. Micheli ex. L., Ileodictyon Tul. & C. Tul., and Laternea Turpin. Additional

species were reported by foreign naturalists (e.g, Cunningham 1868, 1871;

Johow 1896; Spegazzini 1921; Zeller 1939; Santesson 1943; Singer 1969).

Reports from Chilean researchers beginning in the twentieth century (Gunkel

1939, Lazo 1983, 2001; Lazo et al. 1977, Garrido 1981, 1986; Parra & Escudero

1994) have increased the number of phalloid species registered in Chile. The

aim of this study is to provide descriptions of species previously unrecorded

from Chile and a key to facilitate identification of all phalloid species recorded

in this country.

Materials & methods

Specimens were treated according to Rossman et al. (1998) and photographed (in and

ex situ) with a Nikon Coolpix P8 (Nikon, Japan). Vocabulary for morphological features

follows Calonge (1998), and colors were coded following Kornerup & Wanscher (1978).

Dried collections were rehydrated in 90% alcohol before examination. Anatomical

characters were observed under a phase contrast microscope (Axiostar plus, Carl

Zeiss, Germany) on glass slides mounts in water, 3% (w/v) KOH with phloxine, and

cotton blue lactophenol. Vouchers were deposited at the herbaria of Museo Nacional de

Historia Natural, Santiago, Chile (SGO), and Universidade Federal do Rio Grande do

Sul, Porto Alegre, Brazil (ICN). Herbaria acronyms follow Thiers (2013).

Taxonomy

Laternea pusilla Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 343, 1868. FIG. 1

IMMATURE BASIDIOMATA epigeous, ovoid to subglobose, <3.0 cm in diam.;

exoperidium yellowish brown (5E4, 5F4), membranous, fragile, endoperidium

white (5A1) to yellowish white (4A2); one or several rhizomorphs attached at

the base. MATURE BASIDIOMATA epigeous, up to 10 cm high; volva with the

same characteristics of the immature basidioma. RECEPTACLE long-elliptic in

profile, formed by three, occasionally four columns that are free at the base and

united above. COLUMNS up to 8 cm high, 0.4-1.4 cm in diam., pastel red (8A5),

reddish gray (8B2), reddish white (8A2) to orange white (6A2), fading towards

the base, pale orange (5A3) when dry, with a central, longitudinal furrow; with

elongated, delicate crests <1.5 cm high, projecting from the columns’ external

margins, especially near the apex; trapezoid in transversal section, multi-

tubular, tubes 8-10, usually not interconnected, with the widest tubes and

openings on the adaxial face. GLEBIFER situated beneath the arms’ junction,

orange red (8A8); gleba confined to the glebifer, mucilaginous, olive brown

(5E7), fetid.

BASIDIOSPORES cylindrical attenuate at one side, 3.75-4.5 x 1.5-2 um,

chlorohyaline, smooth, thin-walled. Votva’s exoperidium formed by

New Laternea and Lysurus records (Chile) ... 47

; Oe

Fic. 1. Laternea pusilla: A. Basidiome (gl = glebifer; re = receptacle; vo = volva) (SGO 163170).

B, C. Basidiomata expanding from the immature forms (SGO 163171). D. Receptacle column in

transverse section. E. Basidiospores (ICN 192157). Scale bars: A-C = 2 cm; D=2 cm; E=5 um.

pseudoparenchymatous hyphae, 14-39 um in diam., with brownish content,

thick-walled, walls yellowish; endoperidium formed by hyaline hyphae, simple

septate, 3-7 um wide, thin-walled. Cotumns formed by pseudoparenchymatous

hyphae, 9-60 um in diam., hyaline, thin to thick-walled, and walls <2.0 um

wide, yellowish to brownish; crests also pseudoparenchymatous.

HaBIT — growing on mossy barks on shady places. In southern Chile, it

is commonly found within Nothofagus forests. It is possible that also occurs

within Nothofagus forests from Argentina.

SPECIMENS EXAMINED — CHILE. REGION XI, Comuna Puerto Aysén, near Lago

Yulton, 27.V1.2008, P. Sandoval-Leiva (SGO 163171); near Lago Los Palos, 14.XII.2009,

P. Sandoval-Leiva (SGO 163170); REGION X, Comuna Queilén, Isla Tranqui, 8.1I.2009,

48 ... Sandoval-Leiva, Henriquez, & Trierveiler-Pereira

P. Sandoval-Leiva (SGO 163169); Chaitén, Parque Pumalin, Sendero de Alerces,

22.X1.2012, L. Trierveiler-Pereira et al. LTP 310 (ICN 192157). REGI6N XIV, Comuna

Mafil, Fundo Llao-Llao, 12.1X.2011, P. Sandoval-Leiva & J. Marquez (SGO 163168).

REMARKS — The most striking feature of L. pusilla is the presence of crests

projecting from the columns’ external margins, usually found on the upper

part of the receptacle. When basidiomata are not fresh, crests may appear

shrunken. The receptacle morphology varies in many aspects: size, column

structure (arched or more or less straight), number of columns (2-4), and

color (ranging from bright red to pale pinkish). Chilean specimens are larger

than those reported from the neotropics, reaching up to 10 cm high. It is also

important to note that sometimes the columns are pitted. Laternea pusilla is

a neotropical species reported from Central and South America (Dring 1980,

Saenz & Nassar 1982, Meijer 2006).

Lloyd (1909) included seven species in Laternea, but Dring (1980) recognized

only two (L. pusilla and L. triscapa). In this study, we recognize a third species,

L. dringii A. Lopez et al. (Lopez et al. 1981).

Laternea triscapa Turpin was earlier reported from Chile (Montagne 1853-54,

Berkeley 1868, Philippi 1869), but we could not confirm the presence of this

species in the country. Santesson (1943) suggested that the Chilean specimens

identified as L. triscapa were misdeterminations of Colonnaria columnata

(Bosc) E. Fisch [= Clathrus columnatus]. Dring (1980) examined Santesson’s

specimens and preferred to keep the name L. triscapa; however, he noticed

that the specimens had some differences from the tropical ones. It is probable

that these Laternea specimens from southern Chile reported by Santesson are

conspecific with L. pusilla. Gamundi & Horak (2002) reported L. triscapa for

the Patagonian forests in Argentina. The photograph presented by these authors

depicts a pinkish receptacle with small crests (which appear shrunken) at the

columns’ margins. It is possible that this photograph also represents L. pusilla.

Lysurus cruciatus (Lepr. & Mont.) Henn., Beibl. Hedwigia 41: 172. 1902. Fig. 2

IMMATURE BASIDIOME subhypogeous, globose to obovoid, soft, up to 4 cm

in diam.; with numerous mycelial cords attached at the base; exoperidium

grayish yellow (1B3), membranous, endoperidium translucent, gelatinous

and rather fluid. MATURE BASIDIOME epigeous, 6.5 cm high. PSEUDOSTIPE

cylindrical, without longitudinal ribs, 6.5 cm x 1.8 cm, yellowish white (1A2),

internally hollow, externally rough and consisting of one to three layers of

intercommunicating tubes, surmounted by 5-8 arms; arms conical, hollow, up

to 3 cm long, sometimes of different lengths in the same basidiome, initially

united or not at the tips, but usually free when mature, externally brownish

orange (6C6), orange white (6A2) at the center; external surface with a whitish,

longitudinal, central, smooth groove; internal surface slightly concave and

New Laternea and Lysurus records (Chile) ... 49

Fic. 2. Lysurus cruciatus (SGO 159558): A. Basidiome (re = receptacle; ps = pseudostipe; vo = volva).

B. Immature forms. C. Basidiome expanding from the immature form. D. Pseudostipe in transverse

section (note the hollow center). E. Basidiospores. Scale bars: A-C = 2 cm; D = 1 cm; E=5 um.

covered by the glebal mass. GLEBA mucilaginous, yellowish brown (5F5), fetid.

Basipiosporss cylindrical to ellipsoid, (3.3-)3.8-4.0(-4.5) x (1.4-)1.7-2.2

um, chlorohyaline, smooth, thin-walled. PseuposTiPE formed by

pseudoparenchymatous hyphae, 29-52 um in diam., hyaline, thick-walled, cell

walls up to 5 um.

SPECIMEN EXAMINED — CHILE. REGION METROPOLITANA, Melipilla, Cholqui, on soil

under Olea europaea L., 28.VII.2010, Alarcon, Henriquez & P. Sandoval-Leiva (SGO

159558).

50 ... Sandoval-Leiva, Henriquez, & Trierveiler-Pereira

REMARKS — The Chilean collection of L. cruciatus matches descriptions given

by Dominguez (1995), Calonge (1998), and Dring (1980). Lysurus cruciatus is

a saprobic species that grows in enriched cultivated and uncultivated soils with

humus or plant debris in shady and humid places. It has also been recorded

in glasshouses (Kreisel 2001). It is a typical alien species, originally described

in French Guiana and actually with a wide distribution with records in

Australia, Africa, North and South America, Europe, and western Asia (Dring

1980). Kreisel (2001) points out that L. cruciatus is unstable in temperate to

Mediterranean climates, where it does not expand and disappears after one or

several years.

Lysurus Fr. is a widespread genus with five species (Kirk et al. 2008). Prior

to this study, there were two records of the genus in Chile: ‘Lysurus? sp,

reported by Spegazzini (1921) based on an immature basidioma; and Lysurus

periphragmoides (Klotzsch) Dring in Pinus radiata plantations (Garrido 1986,

as Simblum sphaerocephalum Schltdl.).

Discussion

Seven phalloid species are now reported from Chile. Most species have

expanded basidiomata, with the exception of Gelopellis macrospora Zeller, a

hypogeous, truffle-like species.

Phallus impudicus L. is the only species of the genus reported from the

country, in the Valdivia province, observed abundantly during several years in

the winter (Gunkel 1939). Gunkel does not cite voucher material, but since a

good photograph of a specimen is presented, we believe that its identification is

correct. Kreisel (1996) characterized P impudicus by a reticulate receptaculum

surface, lack of veil, and a white pseudostipe, volva, and mycelial strand. Since

the species is common in Europe, Gunkel (1939) theorized that spores had

arrived around the 1930's with European boats that frequently moored at the

port.

Clathrus columnatus was reported only once from Chile, based ona collection

by C. Gay (Montagne 1853-54). Dring (1980) also reported collections from

Valdivia, probably referring to the same collection. We were able to analyze

Gay's collection (Herbarium P), but as the specimen is not well preserved, it

was difficult to verify its identification. As previously discussed, Santesson

(1943) suggested that Chilean specimens recorded as Laternea triscapa might

also represent this species. Clathrus columnatus is reported from America,

Africa, and Australasia, so its occurrence in Chile is expected. The species is

characterized by 2-5 robust, spongy, reddish to orange columns free at the base

and fused at the apex. The gleba is spread in the internal portion of the columns

and is not confined to a glebifer.

New Laternea and Lysurus records (Chile) ... 51

Cunningham (1868, 1871) reported a Clathrus or Ileodictyon species found

on sand in Chiloé Island (Quemchi). The material was later examined by Fischer

(1893) and Dring (1980), who identified it as Ileodictyon cibarium Tul. & C.

Tul. [= Clathrus cibarius (Tul. & C. Tul.) E. Fisch.]. Cunningham's collections

are probably the ones mentioned by Lloyd (1909). Spegazzini (1921) reported

the species as Clathrus cibarius (which he considered a variety of C. gracilis),

growing on soil under Persea lingue (Miers ex Bertero) Nees. Ileodictyon

cibarium was also reported by Parra & Escudero (1994) from the Region of

Valparaiso. Parra & Escudero (1994) also reported Clathrus sp. from Cerro

Pajonal, but their descriptions and illustrations lead us to conclude that their

“Clathrus sp, also represents I. cibarium. Lazo et al. (1977) reported Clathrus

spp. in exotic plantations near Pefuelas Lake, and Lazo (1983, 2001) reported

C. cibarium (mistakenly published as C. gracilis, W. Lazo, pers. comm.) in

Pumanque.

Ileodictyon gracile Berk. was reported from Chile many times (Montagne

1853-54, Léveillé 1846, Johow 1896, Philippi 1869). Montagne (1853-54)

clearly stated that he had no voucher of I. gracile in his fungal collection and

that his description was based on Gay’s illustration and field notes. However,

Léveillé (1846) stated that in Herbarium P there was a Chilean collection of

I. gracile made by Gay. Fischer (1891) identified this collection as I. cibarium;

our examination of Gay’s collection at P confirms Fischer's identification.

Singer (1969) recorded I. gracile in Juan Fernandez Archipelago and southern

Chile, stating that it is easily recognized and the only clathroid species native to

the region. However, our observation of a single glebifer in Singer's collection

(#M7200, SGO) suggests that I. gracile probably represents a Laternea species.

Ileodictyon cibarium is similar enough to I. gracile that many authors

had difficulties in separating them (e.g., Fischer 1890). The two species can

be differentiated by the morphology of the receptacle arms (Dring 1980,

Cunningham 1944): in I. cibarium the arms are thicker, concertina-like, and

not noticeably thickened where anastomosed. ‘The difference between the two

species was well illustrated by Lloyd (1909) and Dring (1980). The original

description of I. gracile (Berkeley 1845) shows a very clear illustration of

the type, which allows us to differentiate it from I. cibarium. Basidiomata of

I. cibarium are usually larger and may even reach a diameter of about 40 cm

(Lloyd 1910).

Due to the great similarity between the two species and erroneous

interpretations of Ileodictyon species, many Chilean records of the genus are

misidentifications. The descriptions and illustrations presented in literature

lead us to conclude that I. cibarium is the only Ileodictyon species definitely

known to occur in Chile.

52 ... Sandoval-Leiva, Henriquez, & Trierveiler-Pereira

Key to the phalloid species known to occur in Chile

1. Basidiome hypogeous, sequestrate (truffle-like); exoperidium thick,

Ve LOWS DITO TOM FISH, oP stews 0 acwartts slaw Mic atewas Pe town Be ae owk’c Gelopellis macrospora

1. Basidiome epigeous when mature, expanded; exoperidium thin,

WETATIS LOM Ae VOOM 9b ote ohn sian eh aE eu SMA MMR idl erine nt aati 2

2-Pseudostipe simple cylindrical! -.5. Ac ov, uc elias ee ee eat neha aad al 3

2. Pseudostipe absent; receptacle clathroid or columnar..................20 eee eee 5

Se Receptacleeonicals og 4 tA ea queer go cae gh ier bCwed ad winder gS ete 8 Phallus impudicus

3. Receptacle sphaerical, clathroid or surmounted by short arms ................... 4

4.-Pseudostipe surmounted by shartarims.. is. wines sce waihe era Lysurus cruciatus

4. Pseudostipe bearing an apical clathroid receptacle ....... Lysurus periphragmoides

5. Receptacle clathroid, whitish; becoming detachable from the volva

a Bi + Ig od Ria Tad hai ny eS oy Os Seay wb Fie We ra, WR ETE) OE Tleodictyon cibarium

5. Receptacle ramified into columns, orange, pink to pale red; remaining

BULAC ME HOC Olah So Bera te fest oa ast etna ethene pitas Satta lt cat SE 6

6. Columns massive, reddish to orange, externally smooth; glebal mass

spread internally in the arms; glebifer absent............. Clathrus columnatus

6. Columns delicate, pinkish to pale red, externally with delicate crests;

glebal mass confined to a single glebifer..................... Laternea pusilla

Acknowledgments

Sandoval-Leiva acknowledges Juan Larrain and Waldo Lazo for their help with

literature and comments, and Jorge Cuvertino for his assistance with Herbarium P

collections. Trierveiler-Pereira thanks M. Rajchenberg, V. Katz, and J.M. Baltazar for

helping during fieldwork. The authors also thank the pre-submission reviewers, Dra.

Andrea Irene Romero (CONICET, UBA, Buenos Aires, Argentina), Dr. Eduardo R.

Nouhra (CONICET, IMBIV, Cordoba, Argentina), and Dr. Armando Lopez Ramirez

(Universidad Veracruzana, Xalapa, México) for valuable comments and suggestions.

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MY COTAXON

http://dx.doi.org/10.5248/128.55

Volume 128, pp. 55-62 April-June 2014

New records of cercosporoid fungi from Poland

UrRSZULA SWIDERSKA-BUREK* & WIESEAW MULENKO

Department of Botany and Mycology, Maria Curie-Sklodowska University,

Akademicka 19, PL-20-033 Lublin, Poland

* CORRESPONDENCE TO: urszula.swiderska-burek@poczta.umcs.lublin.pl

ABSTRACT — Collections of four cercosporoid species are reported for the first time from

Poland: Cercospora echii on Echium vulgare, C. moravica on Caltha palustris, C. viburnicola

on Viburnum opulus, and Passalora pastinacae on Pastinaca sativa. Descriptions and

illustrations of the fungi concerned are presented and remarks containing comparison with

related species, distribution, and habitat are provided.

KEY worpDs — anamorphic fungi, hyphomycetes, leaf spot pathogens

Introduction

Cercospora Fresen. and Passalora Fr. are anamorphs of the ascomycetous

genus Mycosphaerella Johanson (Shin & Kim 2001, Crous & Braun 2003, Crous

2009) belonging to the so-called cercosporoid fungi. Species of these genera are

usually phytopathogenic, often causing leaf spots, occasionally hyperparasitic,

and rarely saprobic (Hsieh & Goh 1990, Crous & Braun 2003, Agrios 2005, To-

Anun et al. 2011).

Investigations of cercosporoid fungi from Poland have been carried out in

the recent years, based on studies of field collections and revision of herbarium

material. Thirty-eight Cercospora and thirty-one Passalora species have been

registered in Poland (Swiderska-Burek 2012). The results will be presented in a

planned monograph of cercosporoid fungi of Poland.

Materials & methods

All collections examined are from Poland. The oldest material treated in this paper

was collected in 1926 and is deposited in the herbarium of University of Warsaw.

The other collections, deposited in the herbarium of the Department of Botany and

Mycology in Lublin, are younger and have recently been re-examined.

The host plants infected with cercosporoid fungi were collected primarily from

grasslands, taken to the laboratory, air-dried, and examined by light microscopy. The

material was mounted in cotton blue and gently heated.

56 ... Swiderska-Burek & Mutenko

The monographs of Crous & Braun (2003), Chupp (1954), and Braun (1995) were

used to identify the fungi. Nomenclature of the host plants follows The Plant List

(http://www.theplantlist.org). Geographic regions of Poland follow the division used

in Kondracki (2009). The specimens examined are deposited in the herbarium of the

Department of Botany and Mycology in Lublin (LBL M) and Faculty of Biology in

Warsaw (WA).

Fic. 1. Cercospora echii (LBL M-12168) on Echium vulgare.

A: Leaf spots. B: Conidiophores. C: Conidia.

Scale bars: A = 20 mm; B, C = 10 um. U. Swiderska-Burek del.

Cercospora & Passalora spp. new for Poland X ... 57

Taxonomy

Cercospora echii G. Winter, Hedwigia 23:190. 1884. Fic. 1

Leaf spots circular, 2-4 mm in diam., dark purplish or almost black center

with pale margin. Fruiting amphigenous, mostly epiphyllous; stromata none or

composed of several brown cells. Conidiophores 1-8 in fascicles, olivaceous-

brown, multiseptate, rarely branched, up to 5 times geniculate, 40-142 x 4-5

um. Conidia hyaline, acicular, straight or slightly curved, 3-12-septate, 40-125

x 2.5-3.5(-4) um.

SPECIMEN EXAMINED — POLAND. WOEYNSKA AND MAEE POLESIE UPLAND, Czumow

village near Hrubieszow town, xerothermic slope, on Echium vulgare L. (Boraginaceae),

19 September 1995, A. Zajac (LBL M-12168).

REMARKS: The fungus has been reported on four species of Echium — E.

italicum, E. platagineum, E. tuberculatum, and E. vulgare — from Europe, Asia,

Africa, North America, and Australia. Echium vulgare is distributed in Europe

and southwestern Asia (Meusel et al. 1965-92, Zajac & Zajac 2009). In Europe,

C. echii is known from five countries (Bulgaria, Portugal, Russia, Spain, and

Ukraine; Crous & Braun 2003, Farr & Rossman 2014).

Cercospora echiorum is also known from Echium species, but it has shorter

and slightly wider conidiophores (15-30 x 4-6 um) and conidia (20-55 x 4-5

um) (Chupp 1954). However, only C. echii has been found on Echium vulgare

(Crous & Braun 2003).

Cercospora moravica (Petr.) U. Braun, Cryptog. Bot. 3: 235. 1993. PIGs2

Leaf spots amphigenous, brown, angular, irregular or subcircular, 1-8 mm

in diam., often confluent, margin indefinite, partially vein-limited. Fruiting

amphigenous, whitish, sub-effuse. Conidiophores in loose or dense fascicles,

hyaline, subcylindrical or geniculate-sinuous, straight, usually aseptate, 5.5-25

x 2-4 um. Conidia hyaline, acicular, 1-8-septate, 30-135 x 1-2.5 um.

SPECIMENS EXAMINED — POLAND. NizINA SRODKOWOMAZOWIECKA LOWLAND,

Garwolin town, water-meadow, on Caltha palustris L. (Ranunculaceae), 28 August 1986,

H. Maszkiewicz (LBL M-12169); NiziIvna POLUDNIOWOPODLASKA LOWLAND, Firlej

village near Lublin town, water-meadow, on Caltha palustris, 12 May 1989, E. Kasinska

(LBL M-12170).

REMARKS: This fungus is confined to Caltha palustris and recorded only from

Europe, although its host species is found also in North America and north

Eurasia (Meusel et al. 1965-92, Zajac & Zajac 2009). Cercospora moravica has

been collected in the Czech Republic, Finland, France, Germany, Great Britain,

and Russia on this host (Crous & Braun 2003; Farr & Rossman 2014).

Cercospora palustris Losa was described on Caltha palustris from Spain in

1944, but this is an unresolved name (Crous & Braun 2003).

58 ... Swiderska-Burek & Mutenko

Fic. 2. Cercospora moravica (LBL M-12170) on Caltha palustris.

A: Leaf spots. B: Conidiophores. C: Conidia.

Scale bars: A = 20 mm; B, C = 10 um. U. Swiderska-Burek del.

Cercospora viburnicola W.W. Ray, Mycologia 33: 174. 1941. FIG. 3

Leaf spots angular or irregular, grayish brown with reddish brown

margin. Fruiting amphigenous. Conidiophores mostly in dense fascicles,

pale olivaceous-brown, straight, slightly curved or sinuous, sparingly septate,

Cercospora & Passalora spp. new for Poland X ... 59

Fic. 3. Cercospora viburnicola (WA 029863) on Viburnum opulus.

A: Leaf spots. B: Conidiophores. C: Conidia.

Scale bars: A = 20 mm; B, C = 10 um. U. Swiderska-Burek del.

60 ... Swiderska-Burek & Mutenko

25-60 x 4-5.5(-6) um. Conidia hyaline, acicular or obclavate, straight or

slightly curved, 3-11-septate, 30-90(—205) x 3-5(-5.5) um.

SPECIMEN EXAMINED — POLAND. POJEZIERZE POLUDNIOWOPOMORSKIE LAKELAND,

Jezioro Kadzionka Lake near Bydgoszcz town, on Viburnum opulus L. (Adoxaceae), 25

September 1926, N.N. (WA 029863, as Cercospora sp. on Clematis vitalba).

REMARKS: This fungus has been recorded from only three countries — China,

Korea, and USA — but from many localities and on many Viburnum spp.

Viburnum opulus occurs in northwestern Eurasia (Meusel et al. 1965-92, Zajac

& Zajac 2009). This is the first record of C. viburnicola in Europe (Crous &

Braun 2003, Farr & Rossman 2014).

This species belongs to the complex of Cercospora apii s. lat. (Crous & Braun

2003). Environmental conditions, especially high temperature and humidity,

influence the length of conidiophores and conidia. In the examined material,

some conidia were longer (<205 um) and wider (usually 3-5.5 um).

Six other cercosporoid hyphomycetes occur worldwide on Viburnum:

C. opuli f. brunneola M.T. Lucas & Sousa da Camara, Passalora viburni (Ellis &

Everh.) U. Braun & Crous, Pseudocercospora opuli (Héhn.) U. Braun & Crous,

P. varia (Peck) J.K. Bai & M.Y. Cheng, P. viburni-cylindrici (F.L. Tai) U. Braun,

and P. viburnigena U. Braun & Crous. However, C. viburnicola is the only

Cercospora species reported on V. opulus (Crous & Braun 2003).

Passalora pastinacae (Sacc.) U. Braun, Nova Hedwigia 55: 213. 1992. Fic. 4

Leaf spots angular, vein-limited, 1-3 mm in diam., mostly yellowish green

or dark brown. Conidiophores solitary or 2-11 in fascicles, pale olivaceous,

olivaceous-brown or yellowish brown, usually 1-2 geniculate, septate, not

branched, 12-48 x 5-7 um. Conidia subhyaline, cylindrical to obclavate,

straight or slightly curved, 1-5-septate, 25-75 x 4-7 um.

SPECIMEN EXAMINED: POLAND. NIECKA NIDZIANSKA BasIN, Rezerwat Skorocice

reserve near Busko-Zdroj town, on Pastinaca sativa L. (Apiaceae), 14 September 1979, J.

Romaszewska-Salata (LBL M-12171).

REMARKS: This fungus is known only from Pastinaca sativa; outside Europe it

has been reported from Africa, North America, and Australia. According to

Meusel et al. (1965-92) and Zajac & Zajac (2009), the host plant commonly

occurs in semi-natural and synanthropic (transformed) habitats in Europe, but

P. pastinacae has been found only in four European countries — Austria, Italy,

Russia, and Romania (Crous & Braun 2003, Farr & Rossman 2014).

Cercospora apii is the only other cercosporoid species known to occur

on Pastinaca. Passolora pastinacae differs from C. apii in having shorter,

cylindrical-obclavate, not quite colourless conidia (Chupp 1954, Crous &

Braun 2003).

Cercospora & Passalora spp. new for Poland X... 61

Fic. 4. Passalora pastinacae (LBL M-12171) on Pastinaca sativa.

A: Leaf spots. B: Conidiophores. C: Conidia.

Scale bars: A = 20 mm; B, C = 10 um. U. Swiderska-Burek del.

Acknowledgments

The authors would like to thank Uwe Braun (Halle/Saale Germany) and Hyeon-

Dong Shin (Seoul, Korea) for reviewing manuscript and very valuable comments. We

are also very grateful to Nomenclature Editor Shaun Pennycook for all the very helpful

remarks and suggestions.

Literature cited

Agrios GN. 2005. Plant Pathology. Fifth edition. Academic Press, Inc., San Diego, California.

Braun U. 1995. A monograph of Cercosporella, Ramularia and allied genera (phytopathogenic

hyphomycetes). Vol. 1. IHW-Verlag Eching.

Chupp C. 1954. A monograph of the fungus genus Cercospora. Ithaca, New York.

Crous PW. 2009. Taxonomy and phylogeny of the genus Mycosphaerella and its anamorphs. Fungal

Diversity 38: 1-24.

62 ... Swiderska-Burek & Mutenko

Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs: 1. Names published in Cercospora

and Passalora. CBS Biodiversity Series 1: 1-571.

Farr DF, Rossman AY. 2014. Fungal Databases, Systematic Mycology and Microbiology Laboratory,

ARS, USDA. Retrieved June 5, 2014, from http://nt.ars-grin.gov/fungaldatabases/.

Hsieh WH, Goh TK. 1990. Cercospora and similar fungi from Taiwan. Maw Chang Book Company.

Kondracki J. 2009. Geografia regionalna Polski [Regional Geography of Poland]. Wydawnictwo

Naukowe PWN. Warszawa.

Meusel H, Jager E, Weiner TE. 1965-92. Vergleichende Chorologie der zentraleuropaischen Flora,

Vol. 1-3. Gustav Fisher Verlag, Jena.

Shin HD, Kim JD. 2001. Cercospora and allied genera from Korea. Plant Pathogens from Korea 7.

Swiderska-Burek U. 2012. Cercosporoid fungi of Poland (manuscript of Phd dissertation in Polish).

Maria Curie-Sktodowska University, Lublin.

The Plant List 2010. Version 1. Published on the Internet; http://www.theplantlist.org/ (accessed

21st May 2014).

To-Anun C, Hidayat I, Meeboon J. 2011. Genus Cercospora in Thailand: taxonomy and phylogeny

(with a dichotomous key to species). Plant Pathology & Quarantine 1(1): 11-87.

Zajac M, Zajac A. 2009. Elementy geograficzne rodzimej flory Polski [The Geographical Elements

of Native Flora of Poland]. Instytut Botaniki Uniwersytetu Jagiellonskiego, Krakow.

MY COTAXON

http://dx.doi.org/10.5248/128.63

Volume 128, pp. 63-70 April-June 2014

New records of Phragmidium species from Pakistan

N.S. AFSHAN’*& A.N. KHALID?

™Centre for Undergraduate Studies & *Department of Botany,

University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590, Pakistan

*CORRESPONDENCE TO: pakrust@gmail.com

ABSTRACT — Five Phragmidium species on different rosaceous hosts are described and illustrated.

Phragmidium violaceum is a new record for Pakistan. Aecidial stages of P. mucronatum,

P. rubi-idaei, and P. tuberculatum are re-described following study of their wall characters

with SEM. Potentilla gerardiana represents a new host record for Phragmidium papillatum.

KEY worpDs — Caeoma, Khanspur, Mansehra, Phragmidiaceae

Introduction

The Phragmidiaceae are a family of rust fungi in the order Pucciniales.

Cummins & Hiratsuka (2003) reported that this family contains 14 genera

and 164 species. Its genera are distinguished by their aecial, uredinial, and

telial stages. Species of Phragmidium Link typically infect plant species in the

Rosaceae. Their telia contain teliospores borne singly on often hygroscopic

pedicels and with smooth or (more often) verrucose walls and 1- to several

transversely septate cells with 2-3 pores in each cell. The aecia are usually

Caeoma-type (with catenulate spores) or less often Uredo-type (with spores

borne singly). About 15 Phragmidium species have been described or reported

from Pakistan (Ahmad et al. 1997; Afshan et al. 2011, 2012). In this paper, we

describe and illustrate five Phragmidium species and provide SEM photographs

to illustrate important aeciospore wall characters.

Materials & methods

Freehand sections of infected tissue and spores were mounted in lactophenol and

gently heated to boiling point. The preparations were observed under a Nikon YS 100

microscope and photographed with a Digipro-Labomed and a JSM5910 scanning

electron microscope. Spores and paraphyses were drawn using a Camera Lucida (Ernst

Leitz Wetzlar, Germany). Spores were measured using an ocular micrometer, with at

64 ... Afshan & Khalid

least 25 spores measured for each spore stage. The rusted specimens have been deposited

in the herbarium of the Botany Department, University of the Punjab, Lahore (LAH).

Taxonomy

(g + * <

bow PRE awe

CR L-UOP.

PLATE 1: Phragmidium mucronatum. (A) SEM photograph of an aecium showing aeciospores

surrounded by slightly incurved to erect paraphyses. (B) SEM photograph of aeciospores showing

verrucose wall ornamentation. (C) Aeciospores with paraphyses. (D) Teliospores. Scale bars:

C, D=10 um.

Phragmidium mucronatum (Pers.) Schltdl., Fl. berol. 2: 156 (1824) PiateE 1

SPERMOGONIA not found. AEciIA hypophyllous, on leaves, bright yellow

to orange yellow, rounded or oblong, scattered, sori 0.1-0.4 x 0.09-0.3 mm;

surrounded by paraphyses; PARAPHYSES numerous, slightly incurved to erect,

clavate, hyaline to nearly colorless, 50-70 um long and 12-15 um wide.

AECIOSPORES globose-subglobose or ellipsoid, 16-28 x 23-32 um (mean 19

x 27.7 um); wall 1.5-2 um thick, with obscure, scattered pores, hyaline with

yellowish orange to dark orange contents, densely and distinctly verrucose.

UrepInia hypophyllous, scattered, yellow; UREDINIOSPORES rare and not very

Phragmidium records for Pakistan ... 65

clear; paraphyses curved, clavate. TELIA hypophyllous, scattered, minute, black.

TELIOSPORES cylindrical to ellipsoid or oblong, 3-8 celled, 27-32 x 57-110

um; wall 4-7 um thick, verrucose, dark brown to chestnut brown; apex conical,

5-10 um thick; pedicel long, hyaline to pale yellow.

MATERIAL EXAMINED: PAKISTAN, KHYBER-PAKHTUNKHAWAH, Khaira Gali, 2347 m

a.s.l., on Rosa macrophylla Lindl., stages I + I + III, 19 June 2008, NSA #150 (LAH

NSA1022).

ComMENTs: Phragmidium mucronatum, cosmopolitan on different Rosa

species (Wilson & Henderson 1966; Hiratsuka et al. 1992), was previously

reported from Pakistan on Rosa sp. from Rawalpindi (Ahmad 1956a,b).

Phragmidium papillatum Dietel, Hedwigia 29: 25 (1890) PLATE 2

SPERMOGONIA and AECIA not found. UREDINIA mostly hypophyllous on

leaves, or on the veins and petioles, bright yellow to orange, scattered, rounded

Oks

C R’LSUOP.

PLATE 2: Phragmidium papillatum. (A) SEM photograph of uredinium containing urediniospores.

(B) A urediniospore showing echinulate to verrucose wall ornamentation. (C) Urediniospores.

(D) Paraphyses (E) Teliospores. Scale bars: C-E = 10 um.

66 ... Afshan & Khalid

or oblong, sori 0.09-0.2 x 0.2-0.5 mm; PARAPHYSES numerous, incurved

to suberect, clavate, hyaline to nearly or quite colorless, 50-80 um long and

11-15 um wide. UREDINIOSPORES globose-subglobose or ellipsoid to ovoid,

16-22 x (19-) 21-32 um (mean 19.0 x 24.5 um); wall 1-1.5 um thick, with

2-5 scattered pores, yellow to yellowish orange, echinulate to verrucose, with a

hyaline pedicel. TeL1a hypophyllous, scattered or gregarious, minute, rounded

or irregular, black. TeLIosPorss cylindrical or ellipsoid to oblong, 3-5 celled,

rarely 2-celled, 28-34 x 57-95 um, slightly or not constricted at the septum;

wall 2-5 um thick, smooth, brown to yellowish brown; pedicel long, 6-8 x

30-95 um, hyaline to pale yellow.

MATERIAL EXAMINED: PAKISTAN, AZAD JAMMU & KASHMIR, Neelum valley, Muchal,

3000 m a.s.l., on Potentilla gerardiana Lindl., stage II, 3 Nov. 2006, NSA #30906 (LAH

NSA1024a); KHYBER-PAKHTUNKHAWAH, Mansehra, Oghi, 1122 m a.s.l., on Potentilla

gerardiana, stages II + II, 16 May 2008, NSA #160408 (LAH NSA1024b).

ComMENts: Phragmidium papillatum has been reported on Potentilla

cryptotaeniae Maxim. and P. matsumurae Th. Wolf from Japan and is also

common in USSR, China, and Korea (Hiratsuka et al. 1992). From Pakistan,

it has previously been reported on Potentilla nepalensis Hook. from Kalam

(Ahmad 1956a,b), Swat valley (Ono & Kakishima 1992), Kaghan valley (Ono

1992), and Murree (Kakishima et al. 1993a,b). Potentilla gerardiana is a new

host for this rust fungus.

Phragmidium rubi-idaei (DC.) P. Karst., Bidr. Kann. Finl. Nat. Folk 31: 52 (1879)

PLATE 3

SPERMOGONIA and UREDINIA not found. AEciA hypophyllous, on leaves,

solitary or in groups, bright yellow, scattered, sori 0.2-0.3 x 0.1-0.4 mm;

surrounded by paraphyses, numerous, incurved to suberect, clavate, hyaline to

nearly colorless, 50-70 um long and 10-16 um wide. AEciosPporEs globose-

subglobose or ovoid to obovoid, 14-17 x 15-23 um (mean 15.9 x 17.5 um);

wall 1-1.5 um thick, hyaline to light yellow with yellowish orange contents,

sparsely echinulate, echines produced on a distinct base. TEL1a hypophyllous,

scattered, naked, black. TEL1osPpores cylindrical, 4-10 celled, 28-35 x 70-122

um, slightly or not constricted at the septum; wall 2-5 um thick, verrucose,

yellowish brown to chestnut brown; apex rounded or attenuated, 10-15 um

long, hyaline; pedicel long, 10-15 x 80-150 um, hyaline to pale yellow.

MATERIAL EXAMINED: PAKISTAN, KHYBER-PAKHTUNKHAWAH, Khaira Gali, 2347 m

a.s.l., on Rubus niveus Thunb., stages I + III, 19 Jun. 2008, NSA #153 (LAH NSA1026).

ComMEnts: Phragmidium rubi-idaei is cosmopolitan in temperate regions on

various Rubus species (Wilson & Henderson 1966; Hiratsuka et al. 1992). It has

previously been reported from Pakistan on Rubus idaeus L. from Khanspur

Phragmidium records for Pakistan ... 67

¢ *

- ee ew

~ R_LeuOP.

PLATE 3: Phragmidium rubi-idaei. (A) SEM photograph of aeciospores showing echinulate wall

ornamentation. (B) A closer view of an aeciospore. (C) Aeciospores. (D) Paraphysis (E) Teliospores.

Scale bars: C-E = 10 um.

(Khalid et al. 1993). The aecidial stage of P. rubi-idaei represents a new record

for Pakistan.

Phragmidium tuberculatum Jul. Mill. Ber. dt. bot. Ges. 3: 391 (1885) PLATE 4

SPERMOGONIA and UREDINIA not found. AEciA hypophyllous, on branches,

petioles and leaves, bright yellow to orangish yellow, rounded or oblong,

scattered, sori 0.1-0.3 x 0.09-0.1 mm; PARAPHYSES numerous, incurved to

suberect, clavate or clavate to capitate, hyaline to nearly or quite colorless,

55-70 um long and 8-10 um wide, with apex 15-20 um in diameter.

AECIOSPORES globose to subglobose or ellipsoid, 18-22 x 21-27 um; wall

1-1.5 um thick, with 5-8 scattered pores, hyaline with orange-yellow contents,

densely verrucose, one or shorter, cylindrical verrucae produced on a distinct

base. TELIA hypophyllous, scattered, minute, black. TELrospores ellipsoid to

cylindrical, 3-7-celled; 25-30 x 57-85 um; wall 3-5 um thick, verrucose, dark

brown to chestnut brown; pedicel long, hyaline to pale yellow.

68 ... Afshan & Khalid

Nu jor. \' X368° S@nm> = sameC R L-UOP.

‘

PLaTE 4: Phragmidium tuberculatum. (A) SEM photograph of an aecium. (B) Aeciospores with

incurved paraphyses. (C) A close view of surface ornamentation of aeciospores showing cylindrical

verrucae produced on a distinct base. (D) Aeciospores. (E) Paraphyses. (F) Multicelled teliospores.

Scale bars: D-F= 10 um.

MATERIAL EXAMINED: PAKISTAN, KHYBER-PAKHTUNKHAWAH, Ayubia National

Park, 2135 m a.s.l., on Rosa moschata Herrm., stages I + II, 24 May 2006. NSA #04

(LAH NSA1028).

ComMENtTs: Phragmidium tuberculatum is cosmopolitan and reported on

different Rosa species (Wilson & Henderson 1966; Farr & Rossman 2014); it has

previously been reported from Pakistan on Rosa webbiana Wall. ex Royle and

Rosa sp. from Kaghan valley, Barum valley, Dungagali, and Swat valley (Ahmad

1956a,b; Kakishima et al. 1993a,b: Okane et al. 1992; Ono & Kakishima 1992).

Phragmidium violaceum (Schultz) G. Winter, Hedwigia 19: 54 (1880) PLATE 5

SPERMOGONIA, UREDINIA and TELIA not found. AECIA amphigenous,

yellowish orange, scattered, on veins and petioles, sori 0.5 x 0.09-0.1 mm.

Phragmidium records for Pakistan ... 69

o _ Heer

CR LOUGP, hele 2 2okU mete aad

om "

PLaTE 5: Phragmidium violaceum. (A) SEM photograph of aeciospores. (B) A closer view of an

aeciospore showing verrucose wall ornamentation. (C) Aeciospores. (D) Paraphyses. Scale bars:

C= 10 um; D = 22 um.

AECIOSPORES globose to ovoid or ellipsoid; wall aculeate-verrucose to

echinulate, verrucae produced on a distinct base, yellow to orange yellow,

sometimes with orangish yellow granules; 16-24 x 20-33 um; surrounded by

peripheral, straight or incurved, clavate and hyaline paraphyses, 94-118 um

long, 8-19um thick, with uniformly thin walls.

MATERIAL EXAMINED: PAKISTAN, AZAD JAMMU & KasHMIR, Neelum valley, Muchal,

3000 m a.s.L, on Rubus ulmifolius Schott, stage I, 3 Nov. 2006, NSA #0101 (LAH

NSA1029a); KHYBER-PAKHTUNKHAWAH, Khanspur, 2135 maz.s1., on Rubus ulmifolius,

stage I, 24 May 2006, NSA #01 (LAH NSA1029b).

Comments: Phragmidium violaceum is widespread in Britain, Ireland, North

America, Europe, and Africa on different Rubus species (Wilson & Henderson

1966; Laundon 1969) and has been introduced as a biological control agent

in Australia, New Zealand, and Chile. This report represents a new record for

Pakistan.

Acknowledgements

We are sincerely thankful to Dr. Abdul Rehman Niazi (Department of Botany,

University of the Punjab, Lahore, Pakistan) and Dr. Omar Paino Perdomo (Dominican

70 ... Afshan & Khalid

Society of Mycology, Santo Domingo, Dominican Republic) for acting as presubmission

reviewers. We are also thankful to Dr. Shaun Pennycook, Nomenclature Editor, for

reviewing the manuscript critically.

Literature cited

Afshan NS, Khalid AN, Iqbal SH, Niazi AR. 2011. Puccinia species new to Azad Jammu & Kashmir,

Pakistan. Mycotaxon 116: 175-182. http://dx.doi.org/10.5248/116.175

Afshan NS, Khalid AN, Iqbal SH, Niazi AR. 2012. Some additions to the Uredinales (rust fungi) of

Fairy Meadows, Northern areas of Pakistan. Journal of Yeast and Fungal Research 3(5): 65-73.

Ahmad S.1956a. Uredinales of Pakistan. Biologia 2: 29-101.

Ahmad S. 1956b. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1: 1-126.

Ahmad §S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Lahore.

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi. Third ed. The American

Phytopathological Society. APS Press, St. Paul, MN.

Farr DE, Rossman AY. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS,

USDA. http://nt.ars-grin.gov/fungaldatabases/

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba Shuppankai, Tsukuba.

Kakishima M, Izumi O, Ono Y. 1993a. Rust fungi (Uredinales) of Pakistan collected in 1991.

169-179, in: T Nakaike, S Malik (eds). Cryptogamic Flora of Pakistan, vol. 2. National Science

Museum, Tokyo.

Kakishima M, Izumi O, Ono Y. 1993b. Graminicolous rust fungi (Uredinales) from Pakistan.

181-186, in: T Nakaike, S Malik (eds). Cryptogamic Flora of Pakistan, vol. 2. National Science

Museum, Tokyo.

Khalid AN, Iqbal SH, Parveen B. 1993. Rust flora of Pakistan. I. Genus Phragmidium Link, on

Rubus spp. Pakphyton 5: 133-136.

Laundon GF, Rainbow AEF. 1969. Phragmidium violaceum. C.M.I. Descr. Pathog. Fungi Bact. 209:

1-2.

Okane I, Kakishima M, Ono Y. 1992. Uredinales collected in Murree Hills, Pakistan. 185-196, in:

T Nakaike, S Malik (eds). Cryptogamic Flora of Pakistan, vol. 1. National Science Museum,

Tokyo.

Ono Y. 1992. Uredinales collected in the Kaghan Valley, Pakistan. 217-240, in: T Nakaike, S Malik

(eds). Cryptogamic Flora of Pakistan, vol. 1. National Science Museum, Tokyo.

Ono Y, Kakishima M. 1992. Uredinales collected in the Swat Valley, Pakistan. 197-216, in:

T Nakaike, S Malik (eds). Cryptogamic Flora of Pakistan, vol. 1. National Science Museum,

Tokyo.

Wilson M, Henderson DM. 1966. British rust fungi. Cambridge University Press, Cambridge.

MY COTAXON

http://dx.doi.org/10.5248/128.71

Volume 128, pp. 71-78 April-June 2014

Life cycle of Aecidium araliae and its new name

MAKOTO KAKISHIMA*? & Qt WANG?

‘Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun, Jilin Province, 130118, China

Faculty of Life and Environmental Sciences, University of Tsukuba, Ibaraki, 305-8572, Japan

*CORRESPONDENCE TO: Kakishima.makoto.ga@u.tsukuba.ac.jp

ABSTRACT — Field observations and inoculation experiments with the uredinial and telial

stages of a rust on Carex stenostachys var. cuneata clarified that these were conspecific with

the spermagonial and aecial stages of Aecidium araliae on Aralia elata. Based on uredinial

and telial morphologies A. araliae is transferred to Puccinia as P. caricis-araliae nom. nov.

Key worps — Araliaceae, Cyperaceae, Pucciniaceae, rust fungus, taxonomy

Introduction

Aecidium araliae, initially published invalidly (Sawada 1943: lacking a Latin

description), was subsequently validated by Ito & Murayama (1943). This rust

fungus has been known to produce only spermogonial and aecial stages on

plants within the Araliaceae and is widely distributed in Japan, Taiwan, and

China: on Aralia elata (Miq.) Seem. in Japan (Ito & Murayama 1943, Ito 1950,

Hiratsuka 1952, 1960, Kakishima et al. 1986, Harada 1994), Taiwan (Tai 1979,

Hiratsuka & Chen 1991) and China (Tai 1979); on A. decaisneana Hance in

Taiwan (Hiratsuka 1943, Ito & Murayama 1943, Sawada 1943, Ito 1950, Tai

1979, Hiratsuka & Chen 1991); on A. chinensis L. in China (Tai 1979); on

A. spinosa L. in Taiwan (Hiratsuka & Chen 1991); on Agalma lutchuense Nakai

[= Schefflera heptaphylla (L.) Frodin] in Japan (Hiratsuka 1952, 1960,

Shimabukuro 1961, Hiratsuka et al. 1985); and on Textoria trifida (Thunb.)

Nakai ex Honda [= Dendropanax trifidus (Thunb.) Makino ex Hara] in Japan

(Hitatsuka 1952, 1960). Spermogonia and aecia were observed on leaves and

petioles that were usually malformed and enlarged by the infections (Fic. 1A-D)

with the surface of these infected tissues becoming yellow because of abundant

aeciospore production.

72... Kakishima & Wang

In June 1992, the rust was observed in Yamagata Pref., Japan, ina field of Aralia

elata cultivated to harvest its sprouts as an early spring vegetable (Fic. 1A,B). By

midsummer the infection had almost disappeared. As other life stages of the

rust could not be found, it was suspected that the rust was heteroecious and

produced other stages on different host plants around these fields. We surveyed

other rust fungi in the same vicinity and collected some candidates for this

species. Inoculation experiments were carried out to confirm their connection.

Finally, we found uredinial and telial stages on Carex stenostachys var. cuneata

(Ohwi) Ohwi & T. Koyama and confirmed these were the other spore stages of

this species. We present results of inoculation experiments and a taxonomic

treatment of the species, based on morphology.

Materials & methods

Inoculations

Telia on dead leaves of Carex stenostachys var. cuneata were collected at Numasawa,

Higashine-shi, Yamagata Pref., Japan on 30 April 1994 (Fic. 2A,B) and kept in a

refrigerator at 5°C for use in inoculations. The leaves with telia were soaked in running

tap water for several days to induce germination of teliospores (Fic. 2C). Once

numerous basidiospores were produced from teliospores then inoculations to A. elata

were carried out several times from May to June at University of Tsukuba, Tsukuba,

Ibaraki Pref., Japan. For inoculation, leaves with germinating teliospores were cut into

small pieces (ca. 5 mm’) and placed on healthy leaves of A. elata. The A. elata plants were

transferred from the field and grown in clay pods. The inoculated plants were kept in a

moist chamber in darkness at about 20°C for 2 days and then transferred into a growth

cabinet at about 20°C with artificial illumination for observations.

Morphological observations

Dry specimens collected in the field or obtained from inoculations were observed

morphologically by light (LM) and scanning electron microscopy (SEM). The spores

or thin sections of sori were mounted in lactophenol solution on glass slides for LM

observations. For SEM, sori and spores obtained from specimens were attached to

specimen holders by double-sided adhesive tape and coated with gold in high vacuum

with an Eiko IB-3 Ion Coater. They were examined with a Hitachi S-430 SEM operated

at 20kV. All specimens were deposited in the fungal herbarium of the Department of

Botany, National Museum of Nature and Science, Tsukuba, Japan (TNS).

Results & discussion

Life cycle

Seven to eight days after inoculation with basidiospores produced by

teliospores on C. stenostachys var. cuneata, small yellow spots of spermogonia

appeared on the surface of leaves and stems of A. elata. The infected tissues

were malformed and discolored. About 10 days later, cupulate aecia with

aeciospores were produced on the same leaves and stems (FiG. 1C). The results

Puccinia caricis-araliae nom. nov. ... 73

FiGuRE 1. Spermogonial and aecial stages of Puccinia caricis-araliae on Aralia elata. A: Symptoms

of A. elata. B: Aecia produced on malformed stems. C: Aecia with aeciospores. D: Aecia produced

by the inoculation. E: Vertical section of a spermogonium. F: Aeciospores. Scale bars: A = 10 cm;

B=5cm;C=1cm;D=3 cm; E=2 um; F= 20 um.

of inoculations confirmed that the telial stage on C. stenostachys var. cuneata

and the aecial stage on A. elata are produced by the genetically same species.

74 ... Kakishima & Wang

TABLE 1. Comparison of Puccinia species on Carex with two germ pores

at each urediniospore base.

UREDINIOSPORES TELIOSPORES AECIAL HOSTS

SIZE WALL SIZE APICAL

SPECIES

(um) (um) THICKNESS (um)

P carieis-araliae! 17-25.5x Palebrown; = 30.5-47 x 1-13 Aralia elata

; 16-24 echinulate 10-16.5 (Araliaceae)

17-26 x Brown; 32-55 x Circaea erubescens

P iensis * : 5-18

pata ae 13-20 echinulate 10-18 (Onagraceae)

P. pulchella * 20-32 x Pale brown; 27-47 x 713 Viola spp.

(nom. illeg.) 15-23 echinulate 11-19 (Violaceae)

21-29 x Pale brown; 29-45 x

P ; 9-13 ¢

pokegurae 16-21 echinulate 16-21

20-2 Pal ; 29-

P. yokotensis * rns alerewe tae 11-13 2

18-21 echinulate 12-18

REFERENCES— !' This paper; * Kakishima & Sato (1983); *Ono & Kakishima (1981); *Ito (1950)

Morphology

Spermogonia and aecia on A. elata produced by inoculations were

morphologically similar with those of specimens collected in the field. As

the morphological characteristics matched those previously described for

A. araliae (Ito & Murayama 1943, Ito 1950), these spermogonial and aecial

stages were identified as A. araliae.

Telial morphology and the two-celled teliospores confirm that the rust

on C. stenostachys var. cuneata belongs to the genus Puccinia (Cummins &

Hiratsuka 2003). Many Puccinia species have been reported on Carex spp. in

Asia (Ito 1950, Tai 1979, Hiratsuka & Chen 1991, Hiratsuka et al. 1992). They

are separated based on position of urediniospore germ pores, as teliospores

are morphologically variable and similar to each other. The Puccinia on

C. stenostachys var. cuneata collected in Yamagata Prefecture has two germ

pores at the base of the urediniospores. It differs morphologically from four

other species on Carex that have two basal germ pores (TaBLE 1). Two of these

species have recorded aecial hosts, but these are in Onagraceae and Violaceae

(and not Araliaceae). Puccinia species producing spermogonia and aecia on

Aralia have not been reported anywhere in the world. Morphological details of

this species are given below.

Taxonomy

Its telial morphology supports the transfer of A. araliae to Puccinia.

However, as the name P araliae has already been published for a different

species producing telia on Aralia, we propose a replacement name in Puccinia

for Aecidium araliae.

Puccinia caricis-araliae nom. nov. ... 75

FiGurE 2. Uredinial and telial stages of Puccinia caricis-araliae on Carex stenostachys var. cuneata.

A: Telia on C. stenostachys var. cuneata. B: Teliospores. C: Urediniospores. D: Germination of a

teliospore. Scale bars: A = 5 cm; B, C = 10 um; D = 15 um.

Puccinia caricis-araliae Kakish. & Q. Wang, nom. nov. Fics 1-3

MycoBank MB 807046

= Aecidium araliae Sawada ex S. Ito & Muray., Transactions of the Sapporo Natural

History Society 17: 171, 1943, non Puccinia araliae Ellis & Everh. 1891.

Type: Japan, Yamagata Pref., Higashine-shi, on Aralia elata, stages 0, I, May 1992

(Neotype designated here, TNS-F-57726). Japan, Yamagata Pref., Higashine-shi,

Numasawa, on Carex stenostachys var. cuneata, stages I, II, 30 April 1994 (Epitype

designated here, TNS-F-57724).

“Aecidium araliae” Sawada, Journal of the Natural History Society

Taiwan 33: 97, 1943, nom. inval. (no Latin).

SPERMOGONIA epiphyllous, surrounded by discolored lesions, yellow to brown,

flask-shaped (type 7 of Cummins & Hiratsuka 2003). Azcta hypophyllous,

densely grouped, cupulate with peridia, pale yellow. AEclosporEs globose to

subglobose, often angular, 18-35 x 16-24 um, wall 2.0-2.5 um thick, hyaline,

verrucose with large granules. UREDINIA hypophyllous, scattered, erumpent,

76 ... Kakishima & Wang

FiGurE 3. Morphology of Puccinia caricis-araliae observed by SEM. A: Aecia on Aralia elata.

B: Vertical section of an aecium on A. elata. C: Aeciospores with large granules. D: Urediniospore.

E: Vertical section of a telium on Carex stenostachys var. cuneata. Scale bars: A = 200 um; B = 50

um; C = 5 um; D = 3 um; E = 25 um.

brown to pale brown. UREDINIOsPORES obovoid or broadly ellipsoid, 17-—25.5

x 16-24 um (av. 21.0 x 18.5 um), wall ca. 1.5 um thick, hyaline to pale brown,

echinulate, germ pores 2 at base. TELIA hypophyllous, scattered, ellipsoid,

erumpent, dark brown. TELIOsPoREs clavate to oblong with round and obtuse

apices, attenuate towards the base, constricted at the septa, 30.5-47 x 10-16.5

um, wall pale brown to brown, thickened at apex (4-13 um), smooth; pedicel

short, persistent.

Puccinia caricis-araliae nom. nov. ... 77

OTHER SPECIMENS EXAMINED: On Carex stenostachys var. cuneata: JAPAN, YAMAGATA

PREE., Higashine-shi, Numasawa, stage II, 18 June 1994 (TNS-F-57725).

On Aralia elata: JAPAN, GUNMA PRer., Kanra-gun, Nanmoku-mura, stages 0, I,

May 2002 (TNS-F-57728); IBARAKI PREF., Tsukuba-shi, stages 0, I, 18 May 1994

(TNS-F-57727); Tsukuba-shi, University of Tsukuba, stages 0, I, post-inoculation, 13

June 1994 (TNS-F-57729); stages 0, I, post-inoculation, 18 June 1994 (TNS-F-57730).

ComMEnts: The type specimen of A. araliae was not designated when Ito &

Murayama (1943) validated this species, although they described two Aralia

species as host plants. Because their specimens also could not be found in the

herbarium of Hokkaido University, Japan (SAPA), we have selected a neotype

specimen for this species.

Spermogonial and aecial stages of this rust fungus were confirmed by

inoculation only on A. elata. However, several species of Araliaceae have been

reported as host plants of A. araliae, which is widely distributed in Japan,

China, and Taiwan. Therefore, we suspect that other Carex species also serve

as uredinial and telial hosts because C. stenostachys var. cuneata is mainly

distributed in northern area of Honshu Island of Japan.

Acknowledgments

We wish to thank Dr. Eric H.C. McKenzie (Herbarium PDD, Landcare Research,

Auckland, New Zealand) and Dr. C. Denchev (Bulgarian Academy of Sciences, Sofia,

Bulgaria) for critical reading of the manuscript. We are also grateful to Mr. S. Yuki

(Agricultural Experimental Station, Yamagata Pref., Japan) for his kind help in collecting

materials and field survey.

Literature cited

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi, 3“ ed. American Phytopathological

Society, St. Paul, Minnesota.

Harada Y. 1994. Materials for the rust flora of Japan VI. Mycoscience 35: 295-299.

http://dx.doi.org/10.1007/BF02268453

Hiratsuka N. 1943. Uredinales of Formosa. Memoirs of the Tottori Agricultural College 7: 1-90.

Hiratsuka N. 1952. Uredinales of Kyushu. Memoirs of the Faculty of Agriculture, Tokyo University

of Education 1: 1-95.

Hiratsuka N. 1960. A provisional list of Uredinales of Japan proper and the Ryukyou Islands. The

Science Bulletin of the Division of Agriculture, Home Economics and Engineering, University

of Ryukyus 7: 189-314.

Hiratsuka N, Chen ZC. 1991. A list of Uredinales collected from Taiwan. Transactions of the

Mycological Society of Japan 32: 3-22.

Hiratsuka N, Hiratsuka T, Hiratsuka K. 1985. Uredinales of the Ryukyu archipelago. Report of

Tottori Mycological Institute (Japan) 23: 55-103.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y,

Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba-shuppankai, Tsukuba.

Ito S. 1950. Mycological flora of Japan, vol. 2, no 3. Yokendo, Tokyo.

Ito $, Murayama D. 1943. Notae mycologicae Asiae Orientalis IV. Transactions of the Sapporo

Natural History Society 17: 160-172.

78 ... Kakishima & Wang

Kakishima M, Sato S. 1983. Puccinia kawakamiensis, a new caricicolous rust, produces the aecial

state on Circaea erubescens. Transactions of the Mycological Society of Japan 24: 403-408.

Kakishima M, Yamaoka Y, Sato S. 1986. Rust fungi (Uredinales) collected from the Tsukuba

botanical garden (1). Annals of the Tsukuba Botanical Garden 4: 43-59.

Ono Y, Kakishima M. 1983. Puccinia pulchella: a new Viola-Carex rust from Japan. Canadian

Journal of Botany 59: 1543-1546. http://dx.doi.org/10.1139/b81-213

Sawada K. 1943. Materials of Formosan fungi 52. Transactions of the Natural History Society

Formosa 33: 96-100.

Shimabukuro S. 1961. Flora of rust fungi in the Ryukyu archipelago. The Science Bulletin of the

Division of Agriculture, Home Economics and Engineering, University of Ryukyus 8: 1-142.

Tai FL. 1979. Sylloge fungorum sinicorum. Science Press, Beijing.

MY COTAXON

http://dx.doi.org/10.5248/128.79

Volume 128, pp. 79-82 April-June 2014

Two rare myxomycete species from the Iberian Peninsula

OscAR REQUEJO' & NICANOR FLORO ANDRES RODRIGUEZ’

' San Xurxo, A Laxe 12B Salceda de Caselas E-36470,

Pontevedra. Spain

? Canovas del Castillo, 1, Vigo E-36202,

Pontevedra. Spain

* CORRESPONDENCE TO: oscarequejo@hotmail.com

ABSTRACT — Two uncommon myxomycete species, Diderma sauteri and Stemonitopsis

microspora, recently recorded in the Iberian Peninsula, are described, illustrated, and

discussed.

Key worps — Amoebozoa, Physarales, Stemonitales, taxonomy, distribution

Introduction

As a result of a fungal and myxomycete inventory of the ‘Rio Caselas’ region,

located in the Northwest of Spain, Pontevedra province (Requejo 2013; Requejo

& Andrés Rodriguez 2013), two interesting taxa were obtained. After a detailed

morphological study, these collections were identified as Diderma sauteri and

Stemonitopsis microspora, two species that are rare in southwestern Europe and

which are described and discussed here.

Materials & methods

The collection and study of the specimens was made following the usual

methodology for myxomycetes (Martin & Alexopoulos 1969; Nannenga-Bremekamp

1991). Macroscopic characters were studied using a stereomicroscopic Nikon model

SMZ745T. Microscopic observations were made using a Nikon E100 microscope.

Samples were prepared in KOH (5-10%). For measurements and microscopic images

of the structures we used the software ProgRes® CapturePro 2.7, associated with the

digital camera Jenoptic CT3 (Jenoptik AG, Germany).

The studied collections are deposited in the personal herbarium of one of the

authors (req-Fungi) with duplicates in Real Jardin Botanico, C.S.LC. (MA-Fungi).

80 ... Requejo & Andrés Rodriguez

Taxonomy

Diderma sauteri (Rostaf.) T. Macbr., N. Amer. Slime-moulds:103.1899 PLATE 1-1-2

SPOROCARPS gregarious or scattered, sessile, globose to subglobose,

sometimes pulvinate, with flattened areas by mutual pressure, up to 1 mm in

diameter. SPOROTHECA pink, grey, pinkish-brown or brownish. HyPOTHALLUS

undetectable. PERrp1uM double, the external layer, cartilaginous, thin, smooth,

separate from the inner; inner layer membranous, grey or iridescent, with

amorphous calcium carbonate granules of 1-3 um diameter. COLUMELLA

rudimentary, calcareous, often reduced to a reddish-brown basal thickening.

CAPILLITIUM filamentous, limeless, smooth, colourless to violet, branched and

few anastomosed. Spor:s free, black in mass, dark violet-brown in transmitted

light by LM, globose, (n = 62) (11.2-)11.8-15.0(-15.2) um, spinulose.

SPECIMENS EXAMINED — SPAIN: PONTEVEDRA. Salceda de Caselas, A Laxe, Rio

Caselas, 29TNG3676, 90 m, on bryophytes, 10-I-2012, leg. Oscar Requejo (req-Fungi

423; MA-Fungi 86674).

ComMEnts — This collection fits well the D. sauteri descriptions of Poulain

et al. (2011), Ing (1999), and Neubert et al. (1995), although we have recorded

a slightly wider range of spore sizes. Our observations of abnormally large

spores up to 21 um diam. coincide with those of Martin & Alexopoulos (1969);

these authors mentioned the possibility of such abnormalities when there is

an anomalous sporocarp development, and therefore we place no taxonomic

importance on such features.

Diderma sauteri is usually associated with bryophytes. The most similar

species is Diderma ochraceum Hoftm., which has smaller spores and bright

orange, red, or yellow sporothecas (Poulain et al. 2011). Diderma subincarnatum

Kowalski shares with D. sauteri the pinkish colors of the sporotheca but the

spores are slightly larger (13-16 um diam) and usually grow in dead branches

and leaves (Poulain et al. 2011).

This is the first confirmed record of D. sauteri in Spain. We know only a

mention for Portugal in Torrend (1909). Neubert et al. (1995) cite D. sauteri

for Spain although they do not refer to a collection. Although they give the

location as “Extremadura” (Spain), this is probably just a reference to Torrend’s

record for “Estremadura” (Portugal).

Stemonitopsis microspora (Lister) Nann.-Bremek., Nederlandse Myxomyceten: 208.

1975 [“1974”] PLATE 1:3-4

SPOROCARPS in small groups, stipitate, 1.8-3.5 mm total height. sPOROTHECA

cylindrical, subcylindrical or slightly conical with rounded apex, brown, 1.5-2.3

mm long. HyPoTHALLUs shared, brown to black-brown. PERIDIUM evanescent.

STIPE 0.6-1.2 mm long, black, with a broadened base. COLUMELLA reaching

the apex of the sporotheca, concolorous with the stalk. CAPILLITIUM tortuous,

Diderma sauteri & Stemonitopsis microspora in Iberia... 81

aa a Se 7 SS _ ad

Seite | Oe oe Ed Sct ee TE ee e --

PLaTE 1. Diderma sauteri (MA-Fungi 86674). 1. Sporocarps; 2. Echinulate spores. Stemonitopsis

microspora (req-Fungi 500): 3. Sporocarps; 4. Spores and capillitium.

becoming very entangled towards the outer part, branched and anastomosed,

forming a fragmented peripheral net. Spores brown to reddish-brown in

mass; hyaline, pale brown by LM, subglobose or slightly polygonal, (n = 59)

(3.7—)3.9-5.1(-5.2) um diam, reticulate.

SPECIMENS EXAMINED — SPAIN: PONTEVEDRA. Salceda de Caselas, A Picona, Rio

Caselas, 29TNG3676, 66 m, on wood of Pinus pinaster, 4-III-2013, leg. Oscar Requejo

(req-Fungi 500; MA-Fungi 86675).

ComMENTs — ‘The specimen fructified on Pinus wood, although S. microspora

is often recorded in the literature as growing on leaves (Martin & Alexopoulos

1969; Poulain et al. 2011). All the morphological characters fit well with the

description provided by Nannenga-Bremekamp (1975) for the species. The

small spore size of S. microspora distinguishes it from other species such as

Stemonitopsis hyperopta (Meyl.) Nann.-Bremek. (5-7 um diam) and Comatricha

pulchella (C. Bab.) Rostaf. (6.5-9 um diam) (Martin & Alexopoulos 1969;

Poulain et al. 2011).

82 ... Requejo & Andrés Rodriguez

According to the data provided by GBIF (2013) there is only one Iberian

record of S. microspora, in Madrid from 1975. We have found no other

bibliographic records of this species for the Iberian Peninsula. The other species

of Stemonitopsis reported from the Iberian Peninsula are S. aequalis (Peck)

Y. Yamam., S. amoena (Nann.-Bremek.) Nann.-Bremek., S. hyperopta, and

S. typhina (RH. Wigg.) Nann.-Bremek. (Lado 1993; Hernandez-Crespo 2006).

Acknowledgments

The authors are very grateful to Dr. Carlos Lado for his help in the confirmation of

the studied species and to Dr. Alfredo Justo for the initial review and translation of the

text. They also thank M. Luisa Castro (Universidade de Vigo, Spain) and Carlos Lado

(Real Jardin Botanico, CSIC, Madrid) for presubmission review.

Literature cited

GBIF. 2013. GBIF data portal, National Botanic Garden Belgium - Myxomycetes.

http://data.gbif.org/datasets/resource/14353 [accessed 2013-05-29]

Hernandez-Crespo JC. 2006. SIMIL, Sistema de Informacién Micoldgica Ibérica en Linea. Real

Jardin Botanico de Madrid, C.S.I.C. Proyecto Flora Micoldgica Ibérica I-VI (1990-2008).

Ministerio de Educacion y Ciencia, Espana. http://www.rjb.csic.es/fmi/sim.php

Ing B. 1999. The myxomycetes of Britain and Ireland. The Richmond Publishing Co. Ltd. Slough,

England.

Lado C. 1993. Bases corolégicas de Flora Micoldégica Ibérica. Numeros 376-692. Cuadernos de

Trabajo de Flora Micoldgica Ibérica 7: 1-305.

Martin GW, Alexopoulos CJ. 1969. The myxomycetes. Univ. of Iowa Press. Iowa.

Nannenga-Bremekamp NE. 1991. A guide to temperate myxomycetes. Biopress Limited. Bristol.

Neubert H, Nowotny W, Baumann K. 1995. Die Myxomyceten (Band II). Karlheinz Baumann

Verlag. Gomaringen.

Poulain M, Meyer M, Bozonnet J. 2011. Les myxomycétes. Fédération Mycologique et Botanique

Dauphiné-Savoie ED. Sévrier.

Requejo O. 2013. Catalogo micoldéxico (Ascomycota, Basidiomycota) das ribeiras do rio Caselas

(Pontevedra, N.O. da Peninsula Ibérica). Mykes 15: 9-90.

Requejo O, Andrés Rodriguez NE. 2013. Catalogo de Myxomycetes (Protozoa) das ribeiras do rio

Caselas. Mykes 15: 91-107.

Torrend C. 1909. Catalogue raisonné des Myxomycetes du Portugal. Boletim da Sociedade

Portuguesa de Ciencias Naturales 2(1/2): 55-73.

MYCOTAXON

http://dx.doi.org/10.5248/128.

Volume 128, pp. 83-91 April-June 2014

Four lecideoid lichens new to China

LinG Hu ', XIN ZHAO '7, LI-YAN SUN ?, ZUN-TIAN ZHAO ! & Lu-LU ZHANG

' Key Laboratory of Plant Stress Research, College of Life Sciences,

Shandong Normal University, Jinan, 250014, P. R. China

* College of Pharmacy, Taishan Medical University, Taian, 271016, P. R. China;

* Correspondence to: lichenzll@aliyun.com

ABSTRACT — Four lecideoid lichen species — Bryobilimbia ahlesii, B. sanguineoatra, Lecidea

promixta, Porpidia grisea — are reported for the first time from China.

Key worps — Lecideaceae, Asia, taxonomy

Introduction

Lecidea (Lecideaceae) was originally described by Acharius (1803). In the

sense of Zahlbruckner, the genus once represented one of the largest lichen

genera and included about 1200 species (Schmull et al. 2011). Subsequently,

Bryobilimbia, Porpidia, and many other obviously more natural units have

been excluded based on the molecular data and the structure of the ascomata,

especially the nature of the hamathecial tissues, ascus apical structures, and

ascospores (Hertel 1975, 1977; Thomson 1997; Buschbom & Mueller 2004;

Smith et al. 2009; Fryday et al. 2014). Lecidea s. str. is now a medium-sized

genus characterized by a Lecidea-type ascus (Hertel 2006, Smith et al. 2009).

Bryobilimbia is distinguished by a hymenium usually with scattered blue-violet

(K+ aeruginose-green) granules, a Porpidia-type ascus, and ascospores with

a warted perispore or thin gelatinous coat (Fryday et al. 2014). Porpidia is

characterized by strongly branched and anastomosed paraphyses, a Porpidia-

type ascus, and large halonate ascospores (Gowan 1989, Nash et al. 2004, Smith

et al. 2009).

Bryobilimbia includes six species, Lecidea s. str. includes about 100 species

(Kirk et al. 2008), and Porpidia includes about 50 species (Smith et al. 2009;

Osyczka & Olech 2011; Fryday et al. 2014; Fryday & Hertel 2014). In China,

16 Lecidea s. str. and 16 Porpidia species have been reported (Wei 1991; Abbas

& Wu 1998; Aptroot & Seaward 1999; Aptroot 2002; Aptroot & Sparrius

84 ... Hu &al.

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

but no species of Bryobilimbia has yet been reported. During our study of

lecideoid lichens from China, we identified four species new to the country —

Bryobilimbia ahlesii, B. sanguineoatra, Lecidea promixta, and Porpidia grisea.

Materials & methods

The specimens studied are preserved in SDNU (Lichen Section of Botanical

Herbarium, Shandong Normal University) and KUN (Kunming Institute of Botany,

Chinese Academy of Sciences). The morphological and anatomical characters of the

specimens were examined under a stereo-microscope (COIC XTL7045B2) and a

polarizing microscope (Olympus CX41). Thallus and medulla were tested with K (a

10% aqueous solution of potassium hydroxide), C (a saturated solution of aqueous

sodium hypochlorite), I (a 10% aqueous solution of aqueous potassium iodide), and

P (a saturated solution of p-phenylenediamine in 95% ethyl alcohol) for identification.

The lichen substances were identified using standardized thin layer chromatography

techniques (TLC) with system C (Orange et al. 2010). Photos of these lichens were taken

under Olympus SZX16 and BX61 with DP72.

Taxonomic descriptions

Bryobilimbia ahlesii (Hepp) Fryday, Printzen & S. Ekman,

Lichenologist 46(1): 29 (2014) Fic. 1

MorpHoLocy — THALLUuS effuse, thin, irregularly rimose, grey-green;

medulla I-; prothallus lacking. APoTHEcta sessile, 0.45-0.6 mm diam.; disc

reddish brown to brown-black, smooth to slightly convex; true exciple usually

prominent and persistent, the outer edge hyaline to pale brown, inner exciple

and hypothecium brown to blackish brown; epihymenium hyaline to yellowish

brown, K-; hymenium hyaline, 60-80 um tall; subhymenium hyaline to pale

yellowish brown, paraphysis c. 1.4 um wide, apices slightly widened to 2.8-5

um, simple or branched in upper part, slightly anastomosed, without apical

cap or hood. Asci clavate, Porpidia-type; ascospores hyaline, simple, ellipsoid,

12-16(-17.5) x (4-)5-6.25 um, with or without thin gelatinous coat. PYCNIDIA

not observed.

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

hypothecium usually with blue-violet (K+ aeruginose-green) granules. No

lichen substances detected by TLC.

DISTRIBUTION — Bryobilimbia ahlesii has been reported from Europe and

North America (Smith 2009, Halda et al. 2011, Fryday et al. 2014). New to

China.

SPECIMEN EXAMINED: CHINA. HEILONGJIANG, Wuchang, Mt. Datudingzi, alt. 1150 m,

on rock, 21 Aug. 2011, D.F. Jiang 20122253 (SDNU).

Comments — Bryobilimbia ahlesii is closely related to B. sanguineoatra but

differs in wider ascospores and occurring on rock rather than bryophytes.

Lecideoid lichens new to China... 85

Fic. 1. Bryobilimbia ahlesii (Jiang 20122253, SDNU). A: Thallus; B: Apothecia; C: Apothecium

section; D: Paraphyses; E: Aeruginose-green K reaction of granules in hymenium and hypothecium;

F; Amyloid reaction of ascus; G: Ascus and ascospores; H: Ascospores.

Bryobilimbia sanguineoatra (Wulfen) Fryday, Printzen & S. Ekman, Lichenologist

46(1): 31 (2014) Figen?

MorpHoLocy — THALLuS membranous, thin, pale yellow; I-. APOTHECIA

sessile, 0.7—-1.1 mm diam.; disc reddish brown, flat and marginate when young

but soon convex and immarginate. True exciple usually exclude, the outer

edge pale brown, inner exciple dark reddish brown; epihymenium yellowish

brown or almost hyaline; hymenium hyaline, 60-85 um tall; hypothecium

86 ... Hu &al.

Fic. 2. Bryobilimbia sanguineoatra (Wang 14595, KUN). A: Thallus; B: Apothecium section;

C: Exciple and epihymenium without crystals; D: Aeruginose-green K reaction of granules in

hymenium; E: Ascus and ascospores; F: Amyloid reaction of ascus; G: Ascospores; H: Paraphyses.

above yellowish brown, below usually paler; paraphysis 1.3-1.6 um wide, apices

slightly widened to 2.5 um, hyaline, mostly simple. Asci clavate, Porpidia-type;

ascospores hyaline, simple, ellipsoid, (9—)11-14 x 3.5-5(-6) um, perispore not

clearly discernable. Pycnrp1A not observed.

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

hypothecium usually with blue-violet (K+ aeruginose-green) granules. No

lichen substances detected by TLC.

Lecideoid lichens new to China... 87

DISTRIBUTION — Bryobilimbia sanguineoatra has been reported from

Europe, Macaronesia, Asia, Africa, and North America (Smith et al. 2009,

Davydov & Printzen 2012, Fryday et al. 2014). New to China.

SPECIMEN EXAMINED: CHINA. YUNNAN, Zhongdian, Mt. Bitahai, alt. 3400 m, on moss,

21 Sept. 1994, L.S. Wang 14595 (KUN).

Comments — Bryobilimbia sanguineoatra is closely related to B. hypnorum but

differs in having a more readily excluded true exciple and narrower ascospores

that are smooth and never septate; in B. hypnorum, the ascospores are often

C: Exciple without crystals; D: Reddish brown N reaction of epihymenium and exciple; E: Ascus

and ascospores; F: Amyloid reaction of ascus; G: Ascospores; H: Paraphyses.

88 ... Hu &al.

thinly 1(-3)-septate, with finely warted perispore, 10-16(-19) x 4.5-6(-7) um.

Bryobilimbia hypnorum is distributed in Europe, Macaronesia, Africa, Asia,

North America, and the subantarctic islands but is still unknown in China

(Smith et al. 2009, Fryday et al. 2014).

Lecidea promixta Nyl., Abh. naturw. Ver. Bremen 14: 490 (1898) Fic. 3

MorPHoLtocy — THALLUuS lacking or very indistinct, dark grey, not

continuous; hypothallus black; medulla I-. APoTHEcrIA sessile, constrict below

when mature, 0.3-0.55(-0.65) mm diam.; disc black, plane to convex. True

exciple persistent, raised and somewhat swollen, of + radially arranged swollen

hyphae, the outer edge dark brown to black, reddish brown within, without

crystals; epihymenium dark green with somewhat brown, irregularly thick,

12-17 mm tall; hymenium hyaline, 40-55 um tall, I-; hypothecium blackish

brown; paraphyses simple or mainly branched only towards the apex, 2.0-3.5

um wide, the apices capitate and widened to 5.0 um wide. Asct clavate, Lecidea-

type; ascospores hyaline, simple, ellipsoid, 10-12.5(-14.5) x 4-5 um. PYCNIDIA

not observed.

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

exciple N+ reddish brown, K-, C-. No lichen substances detected by TLC.

DISTRIBUTION — Lecidea promixta has been reported from Europe (Hertel

2006, Ertz 2008, Liska et al. 2008). New to China.

SPECIMEN EXAMINED: CHINA. SHAANXI, Baoji, Mt. Taibaishan, alt. 3750 m, on rock, 5

Aug. 2005, S.X. Guo 20127072c (SDNU).

ComMENTs — Lecidea promixta is closely related to L. promiscens, which also

has a dark green or green-brown epihymenium and dark brown hypothecium.

However, L. promiscens contains confluentic acid syndrome, and the medulla is

I+ deeply violet (Smith 2009, Zhang et al. 2012), and in China is distributed in

Yunnan (Wang 00-19783; KUN).

Porpidia grisea Gowan, Bryologist 92(1): 48 (1989) Fic. 4

MorPHOLOGY — THALLUS crustose, subrimose to rimose-areolate, even

to weakly verruculose or rugulose, 0.2-0.7 mm thick, medium gray to pale

greenish gray; medulla I+ violet-black; prothallus continuous between thallus

patches, black, thin; soredia absent. APOTHECIA scattered, soon becoming

sessile, 0.7—1.8(-2.2) mm diam.; disc black, plane to weakly convex, usually

moderately pruinose; pruina grayish white; margin bare, distinct, even to

weakly crenulate. ExcIPLE greenish to brownish black at exciple margin,

dark brown within, 87.5-140 um wide, without crystals, with parallel-radiate

hyphae, hyphae c. 4 um wide; epihymenium brown or brownish yellow, without

crystals; hymenium hyaline, 80-100(-112.5) um tall, I+ blue; subhymenium

25-37.5 um, hypothecium blackish brown; paraphyses strongly anastomosed

Lecideoid lichens new to China... 89

Fic. 4. Porpidia grisea (Zhang 20100626, SDNU). A: Thallus; B: Apothecia; C: Apothecium section;

D: Exciple and epihymenium without crystals; E: Amyloid reaction of ascus; F: Paraphyses;

G: Ascospores; H: Conidia.

and apically branched. Asci clavate, Porpidia-type; ascospores hyaline, simple,

ellipsoid, 15-20(-25) x 7-8.5(-10) um, halonate. Pycnip1a immersed, conidia

bacilliform, 7.5-11.5 x c. 1 um.

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

2’-O-methylmicrophillinic acid and 2’-O-methylperlatolic acid were detected

by ee.

DISTRIBUTION — Porpidia grisea has been reported from Europe and North

America (Nash et al. 2004). New to China.

90 ... Hu &al.

SPECIMEN EXAMINED: CHINA. XIZANG, Ridong, Mt. Qimala, alt. 4600 m, on rock, 26

Sep. 1982, M. Zang 4540 (KUN). YUNNAN, Dali, Mt. Cangshan, alt. 3955 m, on rock, 31

Jul. 2013, L.L. Zhang YNO116, YN0121, YN0135 (SDNU); Deqin, Mt. Baimaxueshan,

alt. 4760 m, on rock, 5 Oct. 2009, L.S. Wang 09-31110 (KUN)); Lijiang, Mt. Laojunshan,

alt. 3800 m, on rock, 5 Nov. 2009, H.Y. Wang 20100624, 20100625, 20127130, L.L. Zhang

20100626 (SDNU).

ComMMENTS — Porpidia grisea grows on siliceous or calcareous rock, it is

closely related to P tuberculosa and P. speirea, all of which belong to the

Porpidia speirea complex, which produces moderately pruinose apothecia,

I+ medulla, and confluentic acid. However, P tuberculosa always has soredia,

and the apothecia rarely seen; Porpidia speirea usually has a chalky white

thallus, and apothecia that remain sunken or very broadly sessile when mature;

it grows only on calcareous rock and in China is distributed in Xinjiang

(Abdulla 920442, 920455a, 920728, 920746; XJU).

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. We also thank Dr.

Alan M. Fryday for helping us identify Bryobilimbia species. 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).

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MY COTAXON

http://dx.doi.org/10.5248/128.93

Volume 128, pp. 93-104 April-June 2014

Muscodor strobelii, a new endophytic species

from South India

VINEET MESHRAM, SANJAI SAXENA’, & NEHA KAPOOR

Department of Biotechnology, Thapar University, Patiala, Punjab 147004 India

* CORRESPONDENCE TO: sanjaibiotech@yahoo.com

AsBstRAcT — Muscodor strobelii is described as a new endophytic species existing in stems

of Cinnamomum zeylanicum from South India based on morphological and molecular

evidences along with volatile organic properties.

KEY worpDs — antimicrobial activity, sterile ascomycete, cinnamon, ITS- rDNA

Introduction

Plants hold an enormous diversity of fungal and bacterial endophytes

within leaf and stem tissues, without any obvious indication of their presence

(Bacon & White 2000, Schulze & Boyle 2005, Strobel 2006, Murali et al. 2007,

Hyde & Soytong 2008, Rodriguez et al. 2009, Peay et al. 2010). Endophytes

have been recognized as a promising resource of bioactive compounds and

phytochemicals, which find applications in medicine, industry, and agriculture

(Strobel & Daisy 2003, Mitchell et al. 2008, Tejesvi et al. 2009, Aly et al. 2010,

Gutierrez et al. 2012).

Eight sterile endophytic fungi emanating distinct musky odours due to

presence of volatile organic compounds (VOCs) have been described as novel

fungi: Muscodor albus from Cinnamomum in Honduras (Worapong et al. 2001);

M. cinnamomi from Cinnamomum in Thailand (Suwannarach et al. 2010);

M. crispans from Ananas in Bolivia (Mitchell et al. 2008); M. fengyangensis

from Actinidia and Pseudotaxus in China (Zhang et al. 2010); M. roseus from

Grevillea in Australia (Worapong et al. 2002); M. sutura from Prestonia in

Colombia (Kudalkar et al. 2012); M. vitigenus from Paullinia in Peru (Daisy et

al. 2002); and M. yucatanensis from Bursera in Mexico (Gonzalez et al. 2009).

Apart from morphological and molecular differences, gas chromatography

and mass spectroscopy (GC/MS) of VOCs have also been used as a tool to

demarcate different species of Muscodor (Strobel et al. 2001, Kudalkar et al.

94 ... Meshram, Saxena, & Kapoor

2012). Muscodor species exhibit biological control properties due to their VOC

admixtures that exhibit antibiotic properties and induce lethal effects on insects

and nematodes.

In this study, we recovered an endophytic fungus (#6610) from the stem

tissue of a cinnamon tree (Cinnamomum zeylanicum) growing in Biligiri

Rangaswamy Temple (BRT) wildlife sanctuary, India. Its cultural, chemical,

and molecular characters are distinct from previously described Muscodor

species, and so we name it here as Muscodor strobelii.

Materials & methods

Fungal isolation

Healthy and mature twigs were collected from Cinnamomum zeylanicum growing

in BRT Wildlife Sanctuary, Karnataka, India, during July 2009. The plant parts were

placed in sterile zip pouches and stored at 4 °C until further use. Fungi were selectively

isolated using VOC stress assay as described by Ezra et al. (2004) using the Muscodor

albus CZ620 type strain (gifted by Professor Gary A. Strobel, Montana State University,

USA). The twigs were cut into 1-2 cm pieces under aseptic conditions, surface sterilized

(with 1% sodium hypochlorite for 5 min, 70% ethanol for 1 min, and 30% ethanol for

30 s), and air dried under a laminar hood. The sterilized plant samples were cut into

5-10 mm segments, placed into a potato dextrose agar (PDA) plate containing a 7-day

old culture of M. albus CZ620, and sealed with parafilm. The plates were incubated

at 24 + 2 °C for a fortnight with a 12 h photoperiod and periodically observed for

mycelial germination. The fungi growing out of the plant tissue were aseptically picked

from the tip and transferred to fresh PDA plates so as to obtain pure isolates. VOCs

produced by M. albus CZ620 would permit growth of only volatile-tolerant fungi,

thus enhancing the probability of finding a new Muscodor isolate or strain. The pure

isolates thus obtained were maintained on PDA slants. Combinations of various media

[Richard’s agar (RA), corn meal agar (CMA), banana leaf agar (BLA), malt extract

agar (MEA)] and growth conditions were tested to stimulate spore production (Guo

et al. 1998). The morphological characters (e.g., colony color, texture, growth pattern,

pigment formation, VOCs) and microscopic structures (e.g., hyphal characteristics,

cellular bodies) were observed (Mitchell et al. 2008). The holotype was deposited as a

metabolically inactive culture (liquid nitrogen) at National Fungal Culture Collection of

India, Agharkar Research Institute, Pune (NFCCI).

Scanning electron microscopy

The morphology of isolate #6610 was observed under the scanning electron

microscope following Ezra et al. (2004). The samples were placed in 2.5% glutaraldehyde

in 0.1M phosphate buffer (pH 7.2) overnight at 4 °C. The next day these were washed

twice with 0.1 M phosphate buffer for 10 minutes each, and then washed for 10 minutes

each in 30%, 50%, 60%, 70%, 80%, 90%, 95% and 100% acetone. The samples were then

dehydrated to critical point using liquid CO,. Gold palladium coating of dehydrated

fungal samples was done using a sputter coater and images were recorded in high

vacuum mode using Zeiss Evo40 scanning electron microscope (Ezra et al. 2004,

Kudalkar et al. 2012).

Muscodor strobelii sp. nov. (India) ... 95

Fungal DNA isolation and amplification

Isolate #6610 was grown on PDA in a 90 mm petri dish for 7 days, after which the

mycelium was harvested, freeze dried in liquid nitrogen, and ground into a fine powder

with pestle and mortar. About 10 mg of powdered mycelium was transferred to 2 ml

microfuge tube and genomic DNA extracted using Wizard’ Genomic DNA purification

kit (Promega, USA) as per manufacturer's protocol.

The internal transcribed spacer (ITS) regions 1 and 2 and 5.88

rDNA regions were amplified using Muscodor specific primers M. albus F

(5’-GGGAGGCTACCCTATAGGGGATAC-3’) and M. albus R_ (5’-cAGGGGCCGGAACCAC

TACAGAGG-3’) (Ezra et al. 2010). The 25 ul reaction mixture for ITS amplification

comprised 1 ul of extracted fungal DNA, 10 uM of each primer, 2.5 mM of dNTP, 25

mM MgCl, and 1.5 U of Taq DNA polymerase in 10 X Taq buffer (Bangalore GeNei).

Thermal cycling conditions for Muscodor specific primers was initial denaturation at 96

°C for 5 min followed by 35 cycles of 95 °C for 45 s, 60 °C for 45 s, 72 °C for 45 s, followed

by final extension at 72 °C for 5 min. The ITS amplicons were examined using a 1.5%

agarose gel under UV light in Bio-Rad Gel documentation System using Quantity-1-D

analysis software. The PCR products were purified with Wizard’ SV gel and PCR clean

up system (Promega, USA) following manufacturer's protocol. The purified products

were directly sequenced (Chromus Biotech, Bangalore) using PCR primers mentioned

above.

Sequence assembly, alignment and phylogenetic analysis

Sequences were assembled using Sequencher ver. 5 (www.genecodes.com) and

compared to GenBank sequences using the BLAST software on the NCBI website. The

ITS sequences were aligned with selected sequences of reference taxa obtained from

BLAST using the Clustal W. Any duplicate sequence types were removed from the data

set prior to phylogenetic analyses. After multiple alignments, phylogenetic analysis

was conducted in MEGA 5.0.5. A phylogeny comparing Muscodor species #6610 with

related taxa was generated by the Neighbour-Joining method (Saitou & Nei 1987). The

evolutionary distances (base substitutions per site) were computed using the Kimura

2-parameter method (Kimura 1980). The rate variation among sites was modeled with a

gamma distribution (shape parameter = 5). Alignment gaps were designated as missing

data. Clade stability was assessed by bootstrap analysis with 1000 bootstrap replicates.

The phylogenetic tree (Fic. 8) was drawn and edited in MEGA 5.0.5.

Qualitative analysis of volatiles

The volatile organic gas mixture emanated by the 10-day old isolate #6610 culture

was entrapped using a solid phase micro-extraction (SPME) syringe with a stable flex

fibre of 50/30 di-vinylbenzene/carboxen on polydimethylsiloxane (Supelco, Sigma

Aldrich) following Ezra & Strobel (2004). The fibre was exposed to the air space above

the fungus for 45 min by placing the SPME syringe after drilling a small hole with the

help of sterile needle. The exposed fibre was injected for 30 s in the Shimadzu QP 2010

+ gas chromatograph with thermal desorption system TD 20. A RTX column (diphenyl

95%, dimethyl polysiloxane 5%) with 30 m x 0.25 mm ID and 0.25 mm DF was used

to separate the fungal volatiles. The column was programmed at 100 °C for 2 minutes

before the temperature was increased to 250 °C for 2 minutes and finally to 300 °C

96 ... Meshram, Saxena, & Kapoor

for 13 minutes. The carrier gas was helium and the initial column head pressure was

94.4 KPa. Data acquisition and processing was done on GCMS solution software. The

compounds obtained after GC/MS analysis was then subtracted from the control plate

consisting only PDA medium. The obtained compounds were then tentatively identified

based on their high quality matching with database of National Institute of Standard and

Technology (NIST) compounds (NIST05) and compared with all reported species of

Muscodor (Ezra et al. 2004, Kudalkar et al. 2012).

Bioassay of VOCs produced by Muscodor strobelii

Antimicrobial spectrum of volatiles produced by Muscodor strobelii (#6610) was

tested by uncomplicated bioassay using 90 mm four-quadrant petri dishes (Ezra et.al.

2004). A divided petri dish was used to prevent movement of any diffusible inhibitory

compound from M. strobelii to the test panel microorganisms through the medium.

One quadrant of the petri plate was inoculated with the mycelial plug of an actively

growing M. strobelii culture and the plates were sealed with parafilm and incubated

for 5 days at 24 + 2 °C for production of the VOCs. Subsequently test bacteria and

yeast were streaked in rest of the quadrants. While testing filamentous pathogenic fungi

3 mm mycelial plugs were inoculated. Correspondingly the control plates comprised

only inoculated test bacteria or fungi and were devoid of isolate #6610 allowing it to

grow normally. The antimicrobial action of VOC was determined by monitoring the

growth of the test organism in the control and test plates. To check the VOC inhibitory

or killing effect after 3 days exposure, the exposed culture plugs were placed on fresh

solid media (PDA, YEPD, MHA) to assess their viability (Mitchell et al. 2010).

Taxonomy

Muscodor strobelii Meshram, S. Saxena & N. Kapoor, sp. nov. Fics 1-7

MycoBank MB802337

Differs from Muscodor roseus by its whitish to pale yellowish-orange mycelium and the

absence of hyphal coils.

Type: India, Karnataka, BRT Wildlife Sanctuary, 11°43’-12°09’N 77°01-15’E as

endophytic fungi from internal tissue of stem of Cinnamomum zeylanicum Blume

(Lauraceae), 12 July 2009, leg. Sanjai Saxena #6610 (Holotype, NFCCI-2907; Gen Bank,

JQ409999).

Erymo.ocy: The epithet strobelii honors Dr. Gary Strobel, Emeritus Professor,

Department of Plant Sciences, Montana State University, Bozeman MT, USA.

TELEOMORPH: Unknown

In nature, the fungus is associated with Cinnamomum zeylanicum and is an

ascomycete with sterile mycelia. The fungal colonies grew slowly over PDA and

MEA media, with colony diameter of 17-18 mm on PDA and 27-29 mm on

MEA when incubated at 24 + 2 °C for 7 days with 12 hours of photoperiod.

Fungal colonies on PDA are initially floccose and whitish but with age turn

pale yellowish-orange. Over PDA it generates hyaline, septate branched hyphae

that are 0.76-1.5 um thick. Hyphae form fused cables that interweave into

fishnet-like structures, which further merge to form zinnia flower- or bud-like

Muscodor strobelii sp. nov. (India) ... 97

Fics. 1-7. Muscodor strobelii (#6610). 1. 30-day old colony on PDA. 2. Ropy appearance of

mycelium after 6-day incubation on malt extract agar. 3. Mycelial arrangement of 10-day old culture

on PDA (SEM). 4.Fishnet-like structures (SEM). 5-6. Zinnia flower- and bud-like structures after

10 days on PDA (SEM). 7. Enlarged view of zinnia flower-like structure (SEM). Bars: 1-2 = 2 cm;

3-7 = 10 um.

structures (5.5-15.8 um). The hyphal fabrication is denser in MEA, where the

isolate produces interwoven mycelia that form a 0.76-2.04 um diam. rope-like

structure. The isolate did not produce any fruiting or conidial structure under

tested in vitro conditions.

This M. strobelii isolate was found viable against the VOCs produced by

M. albus CZ620 in the VOC stress bioassay on repeated exposure.

Detailed morphological studies using scanning electron microscopy

The mycelium of M. strobelii in SEM studies exhibits typical characters of a

Muscodor species in forming rope-like strands that branch at a specific angle

(Frc. 3). The cable-like mycelium further forms an interwoven ropy structure

much like fishing net (Fic. 4) that culminates to form a unique structures at its

terminal end to resemble a common zinnia flower- (as seen from the top) or

bud-like (as seen from the side) structure (FIGs. 5-7).

As these structures do not germinate further or sporulate, they should not

be considered to be fruiting bodies. Morphologically, M. strobelii differs from

M. crispans and M. cinnamomi, which exhibit cauliflower-like sterile structures

and possess ropy coiled mycelia. Muscodor yucatanensis has a ropy structure

with swollen hyphae while M. albus exhibits only a ropy mycelium, but both

lack the sterile structures produced by M. strobelii. Muscodor roseus, with its

dense rose coloured mycelium with a variable hyphal thickness that forms

unique hyphal coiled structures, is remarkably dissimilar from M. strobelii.

98 ... Meshram, Saxena, & Kapoor

r AY034665 M. roseus A3-5

JN426991 Muscodor sp. AB2011

GQ220337 M. albus $13-1-2

EU977236 Fungal endophyte P912B

EU195297 M. crispans B23

AY927993 M. albus

AF324336 M. albus CZ620

favs M. albus KN27

Group A

AY527045 M. albus TP21

BL AY527048 M. albus GP206

GQ848369 M. cinnamomi CMU Cib461

JQ409999 Muscodor strobelii #6610

Se FJ664551 Muscodor sp. WG 2009a

FJ917287 M. yucatanensis B110

EU687035 Fungal endophyte 2161

AY 100022 M. vitigenus

53 Group B

EU686979 Fungal endophyte 1896

EU977197 Fungal endophyte P1813B

FJ612989 Fungal sp. ARIZ B342

JF938595 M. sutura SR 2011

HM034856 M. fengyangensis ZJLQO023

100 HM034852 M. fengyangensis ZJLQ151 Group C

86 | HM034853 M. fengyangensis ZJLQ070

AY605713 Hypocrea lixii CBS 226.95 —] Outgroup

0.05

Fic. 8. Phylogeny of Muscodor spp.: Neighbor Joining tree based on ITS rDNA region. ‘The optimal

tree with the sum of branch length = 0.59173282 is shown. There were a total of 632 positions in

the final dataset.

Molecular phylogeny of M. strobelii

The advanced BLAST search of the partial ITS1-5.8S-ITS2 rDNA

region of Muscodor strobelii, which revealed a 94% sequence similarity with

M. cinnamomi (GQ848369; Suwannarach et al. 2010), M. crispans B23

(EU195297; Mitchell et al. 2008), M. albus CZ620 (Worapong et al. 2001),

M. albus E6, and M. albus GP206 and a 92% sequence similarity with M.

yucatanensis, indicates that the isolate belongs to Muscodor genus (TABLE 1).

Representative ITS accessions of Muscodor species and unknown fungal

endophytes (TABLE 1) were analyzed to establish possible phylogenetic

relationships with M. strobelii through an alignment matrix using Clustal W,

MEGA 5.0.5 analysis. The resultant phylogenetic tree divided into 3 major

groups (Group A, B and Group C). Group A clustered M. crispans, M. roseus,

M. cinnamomi CMU-Cib46, M. albus CZ620, 6 strains of M. albus, and one

unknown fungal endophyte 912B with 91% bootstrap support; our isolate was

found to be basal to group A with 100% bootstrap support. Group B comprised

M. yucatanensis, M. vitigenus, M. sutura, Muscodor sp. WG2009a, and 4

unknown fungal endophytes. Muscodor fengyangensis species forms a separate

group (Group C) with 100% bootstrap support; Hypocrea lixii CBS 226.95 was

Muscodor strobelii sp. nov. (India) ... 99

TABLE 1. Advanced ITS-5.8 rDNA gene BLAST search homology analysis

of Muscodor strobelii.

SPECIES / VOUCHER GENBANK NO. QUERY COVERAGE SEQUENCE SIMILARITY

M. cinnamomi CMU Cib-461 GQ848369 98% 94%

M. crispans B23 EU195297 98% 94%

M. albus CZ620 AF324336 98% 94%

M. roseus A3-5 AY034665 98% 94%

M. sutura CA22-D JF938595 91% 91%

M. vitigenus AY 100022 91% 91%

M. yucatanensis FJ917287 90% 92%

M. fengyangensis ZJLQ070 HM034853 90% 90%

M. albus §13-1-2 GQ220337 98% 94%

M. albus I-41.3s AY927993 98% 94%

M. albus TP21 AY527045 98% 94%

M. albus KN27 AY527046 98% 94%

M. albus GP206 AY527048 98% 94%

Muscodor sp. WG2009a FJ664561 90% 92%

Fungal endophyte 912B EU977236 98% 94%

Fungal endophyte 2161 EU687035 90% 92%

Fungal endophyte 1896 EU686979 91% 100%

Fungal endophyte P1813B EU977197 91% 91%

Fungal sp. ARIZ B342 FJ612989 91% 91%

included as outgroup (Fic. 8). These data delineate Muscodor strobelii from the

already described Muscodor species and highlight its novelty.

Bioassay of Muscodor species VOCs against human and plant pathogens

The volatiles emanated by M. strobelii #6610 exhibited a broad spectrum of

activity against yeasts, bacteria, and filamentous fungi. Complete inhibition was

observed in clinical isolates of Escherichia coli, Pseudomonas aeruginosa, and

Staphylococcus aureus after three days of exposure. The VOCs were bactericidal,

as the test bacteria had lost their viability when they were re-inoculated on

fresh medium. The test Candida isolates also showed complete inhibition when

exposed to the M. strobelii VOC mixture and lost their viability.

Among the filamentous fungi, only Penicillium citreonigrum, Botrytis

cinerea, and Aspergillus japonicus were completely inhibited by the VOC

followed by Cercospora beticola and Mycosphaerella fijiensis, inhibited by

60-70%. The completely inhibited fungi were non-viable, whereas Cercospora

beticola, Rhizoctonia solani, and Colletotrichum gloeosporioides showed delayed

100 ... Meshram, Saxena, & Kapoor

TABLE 2. Antimicrobial activity of volatile organic compounds (VOCs) produced by

Muscodor strobelii after 3 days exposure.

TEST CULTURES REPOSITORY rons

(3 days exposure)

Bacillus subtilis MTCC 441 70.3 £1.5

Escherichia coli GMCP 0

Pseudomonas aeruginosa GMCP 0

Staphylococcus aureus GMCP 0

Candida albicans JNU 0

Candida albicans MTCC 3019 0

Candida albicans MTCC 183 0

Penicillium citreonigrum MTCC 160 0

Botrytis cinerea MTCC 359 0

Aspergillus japonicus MTCC 1975 0

Mycosphaerella fijiensis MSU 43 + 2.64

Cercospora beticola MSU 32+2

Rhizoctonia solani MTCC 4634 60.7 + 1.15

Colletotrichum gloeosporioides MTCC 9623 63 +2

Fusarium oxysporum DBTES, TU 70 +4

Lasiodiplodia theobromae DBTES, TU 60+4

*The growth pattern was compared with that of the control plate where the test bacteria, yeast, and

filamentous fungi grew under no stress conditions. All tests were performed in triplicate; their

mean + SD was calculated.

Repositories:

DBTES, TU: Department of Biotechnology and Environmental Sciences, Thapar University, Patiala.

JNU: Jawaharlal Nehru University, New Delhi.

GMCP: Government Medical College, Patiala.

MSU: Montana State University, USA.

sporulation compared to the control (TABLE 2). The M. strobelii VOCs are active

on fungi as well as bacteria. Muscodor albus, M. crispans, and M. fengyangensis

also exhibit antibacterial as well as antifungal activity; however differential

activity exists among the tested bacterial and fungal isolates, which might be

attributed to the chemistry and quantity of the volatiles being produced.

Volatile compounds of Muscodor strobelii

A 10-day old culture of M. strobelii #6610 produced at least 14 volatile

moieties that could be possibly identified based on GC/MS comparison

of authentic standards obtained from commercial sources as well as by

organic synthesis. Primarily the compounds were identified based on their

mass spectral properties when compared to the NIST database. Of all the

compounds produced, 4-octadecylmorpholine was the most abundant, with

the highest peak area (12.5%). The other unique volatile compounds produced

were Tetraoxapropellan, Aspidofractinine-3-methanol, 1-(3,5-di-tert-butyl-

Muscodor strobelii sp. nov. (India) ... 101

TABLE 3. Volatile compounds’ produced by Muscodor strobelii.

RETENTION MOLECULAR Mass

POSSIBLE NAME

TIME FORMULA (Da)

3.03 1.76 1-methyl-4-(prop-1-en-2-yl)cyclohex-1-ene C,H. 136.23

3.69 4.56 Terpinolene Oral: Fe 136.23

13.13 3.5 2-morpholinoethanamine C.H,,N,O 130.18

15.06 8.57 1-(3,5-di-tert-butyl-4-hydroxyphenyl)propan- C,.H,,O, 262.38

l-on

15.33 9.35 Tetraoxapropellan CHO, 200.24

15.92 8.07 Viridiflorol C,,H,,O 222

16.30 4.11 3,5-di-tert-butylcyclohexa-3,5-diene-1,2-dione Cage = x OF 220.30

16.94 8.04 2-(6-tert-butyl-1,1-dimethyl-2,3-dihydro-IH- - C,H,,0, 260

inden-4-yl)acetic acid

17.22 2.96 4-(2-morpholinoethyl)-4H-1,2,4-triazole-3- C,H, .N,O, 225.25

carboxamide

20.193 1.08 1,2,3,4-tetramethyl-5,6-dineopentylbenzene Cry 274.48

21.178 12.53 4-octadecylmorpholine C,,H,,NO 339.6

24.798 3.62 octadec-9-enoic acid C,,H,,0, 282.5

32.054 9.23 Aspidofractinine-3-methanol C,,H,,N,O 310.4

32.385 2.15 (2E)-2-(3-methoxy-5-methyl benzylidene)-7- CaH,.O, 278

methyl-1-indanone

' After 10 days incubation at 24 + 2 °C on potato dextrose agar (PDA) using solid-phase micro-extraction

(SPME) fibre and GC/MS analysis

4-hydroxyphenyl)propan-1-one, viridiflorol, terpinolene, 2-(6-tert-butyl-1,1-

dimethyl-2,3-dihydro-1H-inden-4-yl)acetic acid, 3,5-di-tert-butylcyclohexa-

3,5-diene-1,2-dione, and 2-morpholinoethanamine (TABLE 3). The volatiles

commonly produced by other Muscodor species consist of propanoic acid,

2-methylesters, azulene and naphthalene derivatives, and thujopsene. Muscodor

strobelii produces volatiles belonging to ketones, amines, carboxylic acids, and

alcohols; they are unique and have not been reported from any other Muscodor

species.

Conclusions

As Muscodor species are sterile and do not produce any reproductive

structures, critical examination of cultural, physiological, and biochemical

properties along with molecular phylogenetic data is essential for verifying them

as distinct species. Muscodor strobelii exhibited a variety of common features

shared by Muscodor species; it possessed, however, some unique features like

differences at the molecular level, unique morphological features seen through

scanning electron microscopy, differing antimicrobial susceptibility data,

and a unique volatile gas mixture composition (TABLE 4). Thus M. strobelii is

introduced as a novel species that possesses potential as a mycofumigant.

102 ... Meshram, Saxena, & Kapoor

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Muscodor strobelii sp. nov. (India) ... 103

Acknowledgements

The authors thank the National Biodiversity Development Board, Department of

Biotechnology (NBDB-DBT), New Delhi, India for providing financial support under

project no. BT/PR/10083/NDB/52/95/2007. They express their gratitude to Prof. Dr.

Gary A. Strobel (Professor Emeritus, Montana State University, Bozeman MT, USA) for

providing the M. albus CZ620 type strain, fungal pathogens, and valuable suggestions.

We appreciate the kind help of Dr. Ruchita Pal and Mr. Ajay Kumar (Advanced

Instrumentation and Research Facilities (AIRF), JNU, New Delhi) for SEM and GC/MS

analysis. We are also grateful to Prof. Strobel, Dr. Sanjay K Singh (Agharkar Research

Institute, Pune, Maharashtra, India), and Dr. G.S. Dhingra (Punjabi University, Patiala,

Punjab, India) for presubmission reviews.

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MYCOTAXON

http://dx.doi.org/10.5248/128.105

Volume 128, pp. 105-111 April-June 2014

New record of Setomelanomma holmii on Picea crassifolia in China

based on morphological and molecular data

ZI-QIANG Wu’, XIN-LEI FAN’, TING YANG’, CHENG-MING TIAN’,

YING-MeEt! LIANG? , YAN-FANG Ma}, & SAN-LIN ZHANG?

' College of Forestry, Beijing Forestry University, Beijing 100083, China

? Museum of Beijing Forestry University, Beijing 100083, China

> Linxia Management & Quarantine Station of Forest Pest and Diseases, Gansu 731100, China

* CORRESPONDENCE TO: liangym@bjfu.edu.cn; liangym812@126.com

ABSTRACT — Setomelanomma holmii (Phaeosphaeriaceae) was collected for the first time

in China and for the first time on Picea crassifolia. This represents a new generic record

for Chinese mycobiota. The fungus is identified based on morphological and molecular

characters. An illustrated description of the species is provided.

Key worps — ITS, SNEED, SSU, taxonomy

Introduction

During an investigation of fungal diversity of Picea crassifolia Kom. in

different areas of Kangle County, Gansu Province, China, several samples were

recovered with the morphological characteristics of Setomelanomma holmii.

A literature survey indicated that the genus had not been previously reported

from China (Tai 1979, Qiao et al. 2011, Zhang et al. 2012, Xu et al. 2013).

Setomelanomma M. Morelet is a monotypic genus typified by S. holmii,

which was initially reported in France (Morelet 1980). Later, morphological

studies and phylogenetic analysis inferred from SSU nrRNA regions indicated

that S. holmii occurs also in North America (Canada, United States) and

possibly South Korea (Rossman et al. 2002, Zhang et al. 2009, Kim et al. 2011,

Plewa et al. 2012).

Materials & methods

MORPHOLOGICAL STUDIES. — The specimens and pure cultures were deposited at the

herbarium of Museum of Beijing Forestry University, Beijing Forestry University (BJFC).

Photographs of features were taken using a Nikon P500 digital camera (Nikon, Tokyo).

106 ... Wu &al.

The microscopic procedures follow Rossman et al. (2002). Microscopic observations

and measurements were made from the slide preparations and photographs taken using

a Leica 4D07 microscope (Leica, German). Spore sizes are presented as the range + 5%,

and n = number of spores measured. Special color terms follow Petersen (1996).

MOLECULAR PROCEDURES AND PHYLOGENETIC ANALYSES. — Genomic DNA

was extracted from pure cultures grown in potato dextrose agar (PDA) at 25°C for 2

weeks. A Wizard Genomic DNA Purification Kit (Finnzymes) procedure was used to

extract total genomic DNA from the fruitbodies for polymerase chain reaction (PCR).

DNA sequencing was performed at Beijing Genomics Institute. The nITS region was

amplified with primer pairs ITS1 and ITS4 (White et al. 1990) and the nSSU region

with primer pairs NS1 and NS4 (http://www. biology.duke.edu/fungi/mycolab/primers.

htm). We submitted our newly generated sequences to GenBank as Setomelanomma

holmii (ITS, KF668244; nSSU, KF668245-KF668247) and aligned them with additional

sequences downloaded from GenBank, using BioEdit (Hall 1999) and ClustalX

(Thompson et al. 1997). The sequence alignment was deposited at TreeBase (http://purl.

org/phylo/treebase/; submission ID: 14727). The SSU nrRNA sequences underwent

maximum parsimony analysis using Sordaria fimicola (Roberge ex Desm.) Ces. & De

Not., Xylaria hypoxylon (L.) Grev., and Hypocrea schweinitzii (Fr.) Sacc. as outgroups

(Rossman et al. 2002). The tree was constructed in PAUP* v.4.0b10 (Swofford 2002)

with all characters equally weighted and gaps 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 such as

tree length (TL), consistency index (CI), retention index (RI) and rescaled consistency

index (RC) were calculated for each Maximum Parsimonious Tree (MPT) generated.

MrMODELTEST v.2.3 (Posada & Crandall 1998, Nylander 2004) was used to determine

the best-fit evolution model for each data set for Bayesian inference (BY). Bayesian

inference was calculated with MrBayes v.3.1.2 with a general time reversible (GTR)

model of DNA substitution and a gamma distribution rate variation across sites

(Ronquist & Huelsenbeck 2003). Four Markov chains were run for 2 runs from random

starting trees for 500,000 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.

Taxonomy

Setomelanomma holmii M. Morelet, Bull. Soc. Sci. Nat. Arch. Toulon Var 36: 15.

1980. FIG. 1

Ascomata solitary, scattered, initially immersed, subcuticular, becoming

erumpent, superficial through periderm of small twigs with needles still

attached, perithecioid, 105-279 um diam, black, globose to subglobose, each

with a non-rostrate, periphysate ostiolum, with scattered, sparse to abundant

Setomelanomma holmii on Picea crassifolia (China) ... 107

FiGuRE 1. Setomelanomma holmii. A: Infected twig of Picea crassifolia (BJFC-S425). B—C: Ascomata

on twig (BJFC-S423). D: Longitudinal section of ascomata (BJFC-S425). E-F: Setae on ascomata

(BJFC-S424). G, J: Asci (BJFC-S425). H: Interthecial elements among immature asci (BJFC-S428).

I: Ascospores (BJFC-S425). Scale bars: A = 1 cm; B-C = 100 um; D = 20 um; G-I = 10 um; J = 5 um.

setae; setae short, pale brown to reddish brown, straight to flexuous, slightly

tapered, blunt, 17-54 x 3-6 um. Ascomatal wall 19-36 um thick, of 3-6 layers

of cells that are dark brown and thick-walled toward outside, becoming thin-

108 ... Wu &al.

walled, hyaline toward centrum, wall slightly thicker around non-papillate

ostiolum, of 5-9 layers of cells.

Asci bitunicate, 53-77 x 10-18 um, cylindrical to broadly cylindrical with

rounded apex, with short stalk, attached at base of ascomata, eight-spored.

Interthecial elements arising from basal portion of ascomatal centrum,

multicellular, 2.5-4 um wide, septate, hyaline, anastomosing above asci.

Ascospores broadly ellipsoidal, 16-20.8 x 5.9-8.4 um (ave. 19.7 x 7.4 um,

n = 63), pale to medium brown, three-septate, slightly constricted at median

septum, first and third secondary septa, cells about equal in length, widest

at penultimate cell, apex broadly rounded, base rounded, surface smooth,

gelatinous sheath not observed.

SPECIMENS EXAMINED: CHINA. GANSU PROVINCE, Kangle County, Fucheng, Zhangjian

Village, on living twigs of Picea crassifolia, 17 April 2013, Y.M. Liang (BJFC-S462);

Huguan, Ershilipu Village, on living twigs of Picea crassifolia, 17 April 2013, Y.M.

Liang (BJFC-S463); Minglu, Mingguan Village, on dying twigs of Picea crassifolia 17

April 2013, Y.M. Liang (BJFC-S464, BJFC-S465); Xiajiazhai Nursery, on dying twigs of

Picea crassifolia, 31 July 2013, Y.M. Liang (BJFC-S466; culture BJFC-LSH01; GenBank

KF668244, KF668245); Suji Town, on living twigs of Picea crassifolia, 31 July 2013, Y.M.

Liang (BJFC-S467; culture BJFC-LSH02; GenBank KF668246); Sanchaping Forestry

Farm, on dying twigs of Picea crassifolia, 1 August 2013, Y.M. Liang (BJFC-S468; culture

BJFC-LSH03; GenBank KF668247).

Phylogeny

The SSU nrRNA region dataset included sequences from 44 fungal

specimens representing 40 taxa. The dataset had an aligned length of 1,056

characters of which 836 characters are constant, 86 are variable and parsimony-

uninformative, and 134 are parsimony-informative. Maximum parsimony

analysis yielded 52 equally parsimonious trees (TL = 391, CI = 0.659, RI =

0.833, RC = 0.549). Best model for SSU nrRNA region was estimated and

applied in the Bayesian analysis: GTR+I+G, lset nst = 6, rates = invgamma;

prset statefreqpr = dirichlet (1,1,1,1). The Bayesian analysis resulted in the same

topology with an average standard deviation of split frequencies = 0.008946.

The fungus was readily recognized as a member of Phaeosphaeriaceae

(Pleosporales) based on a primary BLAST search of GenBank using the

rRNA ITS1-5.8S-ITS2 region. The phylogeny inferred from SSU nrDNA

region sequences resulted in seven major clades for the 44 species (Fic. 2).

Three isolates of Setomelanomma holmii formed a strongly supported clade

(MP = 98%; 1.00 BBP = 1.00) with the AF525675 sequence from the S. holmii

holotype strain PC99.4334.

Discussion

Our collections are identified as Setomelanomma holmii based on

morphological observations and molecular phylogenetic analyses.

Setomelanomma holmii on Picea crassifolia (China) ... 109

ree Leptospora rubella AF164361

Ophiobolus fulgidus U43454

— Phaeosphaeria nodorum_AF164372

-- Leptosphaeria doliolum U04205

— Cucurbitaria berberidis U42481

92/1.00

Ophiosphaerella herpotricha U43453 :

Setomelanomma holmii KF668245 Phaecosphaeriaceae/

98/1.00. || |) Setomelanomma holmii KF 668246 Leptosphaericeae

Setomelanomma holmii K¥F668247

Setomelanomma holmii AF525675 (Holotype)

Setomelanomma holmii AF525677

Didymella exigua AF164355

Pseudotrichia aurata AF 164374

Pyrenophora trichostoma U43459

Cochliobolus sativus U42479

Pleospora herbarum U05201 Pleosporaceae

Setosphaeria monoceras AY016352

Setosphaeria rostrata U42487

Herpotrichia juniperi U42483

Melanomma pulvis-pyrius AF 164369

Dangeardiella macrospora AF 164353 Mel t

p91.00 Herpotrichia diffusa U42484 i a hts ae

(— Pleomassaria siparia AF164373

Byssothecium circinans AY016339

87/0.99 |87/0.99 (—Lophiostoma caulium AF 164362

92/1.00

100/1.00

6910.96 |

— Massarina australiensis AF 164362 3

Sahoo Lophiostoma crenatum U42485 Lophiostomataceae

j Massarina bipolaris AF164371

Paraphaeosphaeria michotii AF250817

Paraphaeosphaeria pilleata AF250821

onl Peas winteri AF164371 Montagnulaceae/

TONES r Montagnula opulenta AF 164370 Massariaceae

— Massaria platani AF 164363

80/1.00| -— Keissleriella cladophila AF 164360

——— Leptosphaeria bicolor U04202

Botryosphaeria quercuum AF 154352 .

_{—— Botryosphaeria rhodina U42476 | Botryosphaeriaceae

Botryosphaeria ribis U42477

96/1.00 Hysterium pulicare AF 164358 ‘

| - Hysteropatella clavispora AF164359 | Hysteriaceae

Dothidea insculpta U42474 | Dothideaceae

aria hypoxylon U20378

Hypocrea schweinitzii AF 164357

Sordaria fimicola X69851

Outgroup

FIGURE 2. One of the 52 parsimonious trees illustrating the phylogeny of Setomelanomma holmii

and related species based on nSSU sequences. Parsimony bootstrap proportions greater than 50%

(before the slash marks) and Bayesian posterior probabilities greater than 0.95 (after the slash

marks) are indicated along branches.

Morphologically, they are characterized by setaceous ascomata, bitunicate asci,

and broadly ellipsoidal 3-septate ascospores measured 16-20.8 x 5.9-8.4 um.

Setomelanomma was established as a monotypic genus based on S. holmii

(Morelet 1980) and belongs to the Phaeosphaeriaceae. It can be distinguished

from other genera of Phaeosphaeriaceae by the brown, three-septate ascospores,

setose ascomata, and occurrence on conifers (Barr 1992). The genus is

morphologically similar to Kalmusia Niessl and Phaeosphaeria I. Miyake to

which it appears related (Hedjaroude 1968, Rossman et al. 2002, Zhang et al.

2012). Kalmusia differs in having immersed ascomata at maturity and well-

developed apical papillae (Rossman et al. 2002). Phaeosphaeria is distinguished

from Setomelanomma by absence of setae, causing diseases of angiosperm

110... Wu &al.

hosts, and presence of coelomycetous anamorphs (Holm 1957, Hedjaroude

1968, Leuchtmann 1984).

In the phylogenetic tree (Fic. 2), the three isolates of S. holmii from China

nested within the Phaeosphaeriaceae, and grouped closely with the type strain

(PC99.4344) of S. holmii, forming a well supported lineage distinct from other

species with a 92% bootstrap value and 1.00 Bayesian posterior probability.

Setomelanomma holmii causes a disease called spruce needle drop (SNEED).

It has been reported in association with Picea pungens Engelm. and P. glauca

(Moench) Voss (Morelet 1980; Rossman et al. 2002) in France and North

America. This is the first report of this species in China, and the first report on

the new host P. crassifolia. Although Kim et al. (2011) reported S. holmii from

soybean paste in Korea, this report was based solely on a sequence similarity.

Acknowledgments

We express our gratitude to Dr. Amy Rossman (Systematic Mycology and

Microbiology Laboratory, USDA-ARS, MD, USA) and Dr. Yi-Jian Yao (Institute of

Microbiology of the Chinese Academy of Sciences, Beijing, China) who reviewed the

manuscript. The research was financed by the National Natural Science Foundation of

China (No.31170603).

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Barr ME. 1992. Additions to and notes on the Phaeosphaeriaceae (Pleosporales, Loculoascomycetes).

Mycotaxon 43: 371-400.

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

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

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.

Hedjaroude GA. 1968. Etudes taxonomiques sur les Phaeosphaeria Miyake et leurs formes voisines

(Ascomycetes). Sydowia 22: 57-107.

Holm L. 1957. Etudes taxonomiques sur les Pléosporacées. Symbolae Botanicae Upsalienses 14(3).

188 p.

Kim JY, Yeo SH, Baek SY, Choi HS. 2011. Molecular and morphological identification of fungal

species isolated from Bealmijang meju. Journal of Microbiology and Biotechnology 21:

1270-1279. http://dx.doi.org/10.4014/jmb.1105.05013

Leuchtmann A. 1984. Uber Phaeosphaeria Miyake und andere bitunicate Ascomyceten mit

mehrfach querseptierten Ascosporen. Sydowia 37: 75-194.

Morelet M. 1980. Sur quatre Dothideales. Bulletin de la Société des Sciences Naturelles et

dArchéologie de Toulon et du Var 36: 14-15.

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

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Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve.

Plewa DE, Jossey S, Porter SK, Bissonnette SM. 2012. First report of spruce needle drop

associated with Setomelanomma holmii on spruce in Illinois. Plant Disease 96: 459.

http://dx.doi.org/10.1094/PDIS10110862

Setomelanomma holmii on Picea crassifolia (China) ... 111

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

Qiao Q, Jin J. 2011. Two new Chinese records of Phaeosphaeria. Journal of Qingdao Agricultural

University 28: 44-46.

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

Rossman AY, Farr DF, Castlebury LA, Shoemaker R, Mengistu A. 2002. Setomelanomma holmii

(Pleosporales, Phaeosphaeriaceae) on living spruce twigs in Europe and North America.

Canadian Journal of Botany 80: 1209-1215. http://dx.doi.org/10.1139/b02-111

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

4.0b10. Sinauer Associates, Massachusetts.

Tai FL. 1979. Sylloge fungorum sinicorum. Science Press, Beijing.

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

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

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

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methods and applications. Academic Press, San Diego.

Xu ZL, Feng ZD, Zhao CY, Zheng JH, Yang JJ, Tian FX, Peng HH, Wang C, Peng SZ, Sher H.

2013. The canopy rainfall interception in actual and potential distribution of Qinghai

spruce (Picea crassifolia) forest. Journal of Hydrology and Hydromechanics 61: 64-72.

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Zhang Y, Schoch CL, Fournier J, Crous PW, Gruyter J, Woudenberg JHC, Hirayama K, Tanaka

K, Pointing SB, Spatafora JW, Hyde KD. 2009. Multi-locus phylogeny of Pleosporales: a

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Zhang Y, Crous PW, Schoch CL, Hyde KD. 2012. Pleosporales. Fungal Diversity 53: 1-221.

http://dx.doi.org/10.1007/s13225-011-0117-x

MY COTAXON

http://dx.doi.org/10.5248/128.113

Volume 128, pp. 113-115 April-June 2014

Thalloloma ochroleucum (Graphidaceae),

a new species from Guizhou, China

ZE-FENG JIA? & KLAUS KALB ™?3

"College of Life Sciences, Liaocheng University, Liaocheng, 252059, Shandong Province, China

? Lichenologisches Institut Neumarkt, Im Tal 12, D-92318 Neumarkt, Germany

> University of Regensburg, Institute for Botany,

Universitatsstrafse 31, D-93040 Regensburg, Germany

CORRESPONDENCE TO *: zffia2008@163.com

AsstTRACT — Thalloloma ochroleucum is described as a new species from Guizhou province

in southern China. It is characterized by subimmersed lirellae, open discs with pale orange

pruina, transversely septate hyaline ascospores, and the presence of norstictic acid.

KEY worps — taxonomy, lichenized fungi, Ostropales, Ostropomycetidae

Introduction

In recent years, many systematic and taxonomic changes have occurred in

the Graphidaceae (Ostropales, Ostropomycetidae). Staiger (2002) revised the

spore-based generic system established by Miller Argoviensis (1880, 1882) in

the family and reintroduced several genera with revised concepts, including

Thalloloma Trevis. Thalloloma is characterized by a corticolous thallus,

lirelliform apothecia, uncarbonised proper exciples, and hyaline ascospores

(septate with lenticular locules or muriform), reacting I+ blue. Thalloloma

ochroleucum is described here as new to science.

Materials & methods

The specimens are deposited in the Herbarium Mycologicum Academiae Sinicae

— Lichenes (HMAS-L). A dissecting microscope (TEcH XTS-30D with Canon 600D

camera) and a light microscope (OLympus SZ-51) were used for the morphological

and anatomical studies. Measurements and illustrations were taken from manual cross-

sections of fruit bodies in tap water. Amyloidity of the ascospores was tested using

Lugol's solution. Spot tests with KOH (20%) were performed on the thallus surface and

on thin thallus sections. The lichen substances were detected and identified by thin-

layer chromatography (TLC) (Culberson & Kristensson 1970, Culberson 1972, Orange

et al. 2001).

114... Jia & Kalb

Taxonomy

Thalloloma ochroleucum Z.F. Jia & Kalb, sp. nov. Pu. 1

FUNGAL NAME FN570059

Differs from Thalloloma deplanatum by its production of norstictic acid and its longer

ascospores.

Type: China, Guizhou province, Tongren City, Mt. Fanjing, Daling, 27°55’N 108°41’E,

alt. 1800 m, on bark of Rhododendron rufum Batalin (Ericaceae), 22.VIIL. 1963. coll. J.C.

Wei 0474 (Holotype, HMAS-L 047744).

ETyMOLOGy: Latin ochroleucus, a reference to the ochre-colored disc.

Thallus corticolous, crustose, pale white to yellowish, thin, smooth, tightly

attached to the substratum, lacking isidia and soredia. Apothecia lirelliform,

numerous, short to elongate, open, simple, subimmersed, 0.8—2.5 mm long,

0.3-0.6 mm wide, with thick thalline margin, not striate, scattered over the

thallus, disc opened with orange pruina. Proper exciple inconspicuous,

uncarbonised. Epithecium 8-10 um thick, brown. Hymenium colorless, not

inspersed, 150-180 um high, I-. Hypothecium brownish, 5-15 um high.

Paraphyses simple, 1-2 um wide, apices becoming broad, brownish. Asci

itt as

5 “|

PiaTeE 1. Thalloloma ochroleucum (holotype). A. Thallus on bark. B. Cross section of an apothecium.

C. Asci containing ascospores. D. Immature ascospores in an ascus. E. Mature ascospores in asci.

Scales: A = 1 mm; B = 100 um; C, D = 50 um.

Thalloloma ochroleucum sp. nov. (China) ... 115

cylindrical to clavate, 95-110 x 15-20 um, 8-spored. Ascospores hyaline,

narrowly ellipsoidal, transversely septate, 15—18-locular, 40-80 x 7.5-12 um,

I+ bluish.

CHEMISTRY: C-, K+ yellow to brown, P+ yellow; norstictic acid detected by

TEC:

DISTRIBUTION & ECOLOGY: Thalloloma ochroleucum is present in Mt. Fanjing

(Guizhou province) within a subtropical rainforest in southern China. The

species was found on bark of Rhododendron rufum accompanied by Graphis

spp. such as G. hossei Vain., G. vittata Mull. Arg., and G. librata C. Knight.

ADDITIONAL SPECIMEN EXAMINED: CHINA, GUIZHOU PROVINCE, Tongren City, Mt.

Fanjing, Zhangjia Dam, alt. 1000 m, on bark, 16. VIII. 1963. coll. J.C. Wei 0204 (HMAS-L

047741).

Comments: Thalloloma ochroleucum resembles T: deplanatum (Nyl.) Staiger,

which also has transversely septate ascospores and open discs, but differs by

its shorter thick-walled ascospores, connective paraphyses, and absence of

norstictic acid (Staiger 2002). The new species is readily distinguished from

T. microsporum Z.F. Jia & J.C. Wei described from China, which has red

apothecia and smaller ascospores (15.5—20 x 5.5—8.0 um; Jia & Wei 2009).

Acknowledgments

This research was supported by the National Natural Science Foundation of China

for the funds (No. 31270066 & 31093440). The authors are grateful to Dr. Alan W Archer

and Prof. Jae-Seoun Hur for their valuable comments on the manuscript.

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

Jia ZF, Wei JC. 2009. A new species, Thalloloma microsporum (Graphidaceae, Ostropales,

Ascomycota). Mycotaxon 107: 197-199. http://dx.doi.org/10.5248/107.197

Miller Argoviensis J. 1880. Lichenologische Beitrage 10. Flora 63: 17-45.

Miller Argoviensis J. 1882. Lichenologische Beitrage 15. Flora 65: 291-402.

Orange A, James PW, White J. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London, United Kingdom.

Staiger B. 2002. Die Flechtenfamilie Graphidaceae. Studien in Richtung einer natiirlicheren

Gliederung. Bibliotheca Lichenologica 85: 1-526.

MYCOTAXON

http://dx.doi.org/10.5248/128.117

Volume 128, pp. 117-125 April-June 2014

A new species of Marasmius sect. Sicci from India

ARUN KUMAR DUTTA, SWARNENDU CHANDRA,

PRAKASH PRADHAN, & KRISHNENDU ACHARYA*

Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany,

University College of Science and Agriculture, Taraknath Siksha Prangan,

University of Calcutta, Kolkata-700019, West Bengal, India

* CORRESPONDENCE TO: krish_paper@yahoo.com

ABsTRACT — ‘The marasmioid fungus Marasmius midnapurensis (Marasmiaceae,

Basidiomycota) is described as a new species from India. Analysis of the internal transcribed

spacer-1 region (ITS1) of the nuclear ribosomal RNA gene suggests that M. midnapurensis

is phylogenetically distinct from closely related species and confirms its position within

Marasmius sect. Sicci. Data on macro- and microscopic characters, habitat and comparisons

with morphologically similar species are provided.

Key worps — Agaricomycetes, biodiversity, phylogeny, taxonomy

Introduction

Marasmius Fr. (Marasmiaceae, Agaricomycetes, Basidiomycota), as

traditionally accepted by Singer (1986), is polyphyletic (Wilson & Desjardin

2005). Based on the nLSU rDNA sequences, Wilson & Desjardin (2005)

restricted the genus to a monophyletic lineage containing only sections

Marasmius, Sicci, Hygrometrici, Globulares, Neosessiles, Scotophysini, and

Leveilleani where taxa in Sicci and Globulares form a large joint clade

(Wannathes et al. 2009). Marasmius sect. Sicci includes species characterized

by a hymeniform pileipellis of broom cells of the Siccus-type and dextrinoid

hyphae (Singer 1958, 1986).

Sixty-eight species and one variety have been reported thus far in India

(Manjula 1983, Manimohan & Leelavathy 1989, Bilgrami et al. 1991). In the state

of West Bengal only nine species of the genus have been reported: Marasmius

consocius Berk., M. erythropus (Pers.) Fr., and M. burkillii (Massee) Manjula

from the Darjeeling area (Berkeley 1851, Massee 1910); M. pangerangensis

Henn., M. campanella Holterm., and M. haematocephalus (Mont.) Fr. from

118 ... Dutta & al.

Calcutta (Bose 1949, Bose & Chatterjee 1950, Roy 1953); M. umbrinus Pegler

from Bankura district (Ray & Samajpati 1979); and M. androsaceus (L.) Fr. and

M. siccus (Schwein.) Fr. from the lateritic region of West Bengal (Pradhan et al.

2011). All of these records should be revised in the light of modern taxonomic

concepts and the many new species that have been described, especially from

tropical regions.

The present paper describes Marasmius midnapurensis, a new fungal species

from West Bengal.

Materials & methods

Basidiomata sampling and morphological studies

Basidiocarps of Marasmius midnapurensis were collected in 2011 during field trips

to the state of West Bengal, India. Their morphology and ecology were noted and colour

photographs were taken in the field. Microscopic features were obtained from dried

material by mounting free-hand sections in 5% potassium hydroxide (KOH), Melzer’s

reagent, Congo red, or lactophenol-cotton blue and examination using a Carl Zeiss

AX10 Imager Al phase contrast microscope. Colour terms follow the British Fungus

Flora Colour Chart (Anonymous 1969).The terms used to describe lamellae spacing are

L for number of lamellae and | for number of lamellulae between two lamellae. Spore

statistics include: X_ , the arithmetic mean of the spore length by spore width (+ standard

deviation) for n spores measured in a single specimen; Q, the quotient of spore length

by spore width in any one spore, indicated as a range of variation in n spores measured;

Q _. the mean of Q-values in a single specimen; n, total number of spores measured;

s, the number of specimens. The holotype collection has been deposited in the Calcutta

University Herbarium (CUH).

DNA extraction, Polymerase Chain Reaction, and sequencing

Genomic DNA was extracted from dried (50°C) herbarium tissue (10-50 mg)

using the ‘Fungal gDNA Mini Kit’ (Xcelris Genomics, Ahmedabad, India). The internal

transcribed spacer-1 region (ITS1) was amplified using ITS1-F (Gardes & Bruns 1993)

and ITS2 (White et al. 1990) primer pair. A hot start of 2 min at 94°C was followed by

30 cycles consisting of 30 s at 94°C, 1 min at 56°C, 1 min at 72°C, and a final elongation

step of 5 min at 72°C. PCR products were checked on 2% agarose gel stained with

ethidium bromide. PCR products were purified using QIAquick® Gel Extraction Kit

(QIAGEN, Germany) and sequencing was done using Sanger methods. The obtained

sequence generated was submitted to GenBank.

Phylogenetic analysis

Our new ITS1 sequence was submitted to GenBank (www.ncbi.nlm.nih.gov), and

related Marasmius sequences were identified by a BLASTn search (http://blast.ncbi.

nlm.nih.gov/). Sequences used in the phylogenetic analysis are indicated in Fic. 3, with

Mycena pura (Pers.) P. Kumm. and Marasmius rotula (Scop.) Fr. as outgroup.

The evolutionary history was inferred using the Neighbor-Joining method (Saitou

& Nei 1987). The bootstrap consensus tree inferred from 1000 replicates (Felsenstein

1985) is taken to represent the evolutionary history of the taxa analyzed (Felsenstein

Marasmius midnapurensis sp. nov. (India) ... 119

1985). Branches corresponding to partitions reproduced in less than 50% bootstrap

replicates are collapsed. The percentage of replicate trees in which the associated taxa

clustered together in the bootstrap test (1000 replicates) is shown next to the branches

(Felsenstein 1985). The tree is drawn to scale, with branch lengths in the same units as

those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary

distances were computed using the Jukes-Cantor method (Jukes & Cantor 1969) and

represent the number of base substitutions per site. All positions containing gaps and

missing data were eliminated from the dataset (Complete deletion option). There were

a total of 142 positions in the final dataset. Phylogenetic analyses were conducted in

MEGA4 (Tamura et al. 2007).

Taxonomy

Marasmius midnapurensis A.K. Dutta, P. Pradhan & K. Acharya, sp. nov. Fics 1, 2

MycoBank MB809410

Differs from Marasmius jasminodorus by its lighter colored and striate pileus, creamy

lamellae forked towards the margin, non-strigose stipe base with whitish mycelium,

slightly larger basidiospores, and the absence of a sweetish odor.

Type: India, West Bengal, Midnapur District, Ramnagar, Kasaphaltala, 21°43’29.9”N

87°31'36.6’E, 10 m asl., on dried Acacia leaves and on wood, 11 Aug. 2011, A.K. Dutta,

P. Pradhan & K. Acharya (Holotype, CUH AMT002; GenBank, KF682470).

ETyMOLoGy: specific epithet refers to the type locality.

PILEUS 22-27 mm diam., broadly convex, sometimes umbonate, smooth, viscid

when moist, light brown to light greyish brown with irregular light yellowish

brown patches in the center, hygrophanous, smooth, striate. CONTEXT $0.8 mm

thick, creamy, not changing colour when exposed. LAMELLAE L = 13-14, 1 =

3-4, adnexed, subdistant, forked towards the margin, creamy, regular, <4 mm

broad, spacing <5 mm, margin even, concolorous. COLLARIUM absent. STIPE

53-65 mm long, 2 mm broad overall, central, cylindrical, dark brick brown

lower, creamy in upper part, equal, hollow, cartilaginous, strict to curved at

lower portion, dry, smooth, flesh concolorous with the pileus, non-insititious,

base covered with whitish mycelium. Opor and TasTE mild.

BASIDIOSPORES (10.7—)11.08-12.2(-15) x (3.5-)3.9-4.3(-4.7) um [X_ = 11.57

+ 1.04 x 3.9 + 0.39, Q = 2.5-3.5, Q. = 3 + 0.24, n = 150 spores (30 spores each

among the collected 5 basidiocarps), s = 1 specimen], ellipsoid, slightly curved

in profile, smooth, hyaline, inamyloid, thin-walled. Basrpra (17.9-)23.2-23.6

(-24.4) x 4.7-6.8(-7.9) um, clavate, hyaline, 4-spored, sterigmata 3.2-3.6 um

long. BaAsIDIOLES 17.9-21.5 x 6.8-7.2 wm, fusoid to clavate, hyaline.

CHEILOCYSTIDIA common, of Siccus-type broom cells, main body 13.6-17.9

x 7.2-10 um, cylindrical to clavate, hyaline, inamyloid, thin- to thick-walled,

apical setulae 3.9-6.8(-10) x 1.4-1.8 um, irregular in outline, obtuse to

subacute, yellow to brownish yellow, thick-walled. PLEUROcysTipIA absent.

PILEIPELLIS hymeniform, mottled, composed of Siccus-type broom cells, main

120 ... Dutta & al.

Figure 1. Marasmius midnapurensis (holotype): A. Basidiomes. B. Siccus-type cells of pileipellis.

C. Basidiospores. D. Basidium. E. Cheilocystidia. F. Caulocystidia. Scale bars: A = 10 mm;

B, F= 10 um; C-E =5 um.

body 13.6-14.3 x (6.8-)7.1-10.3(-10.7) um, clavate to broadly clavate, often

branched, hyaline, inamyloid, thin- to thick-walled, apical setulae 3.9-6.8(10) x

1.4-1.8 um, crowded, cylindrical to irregular in outline, obtuse to subacute,

yellowish brown to brown, thick-walled, setae absent. PILEUS TRAMA

interwoven, strongly dextrinoid. LAMELLAR TRAMA hyphae interwoven,

cylindrical to inflated, smooth, hyaline, dextrinoid, thin-walled, non-

gelatinous. STIPITIPELLIS hyphae 6-7 um broad, parallel, yellowish brown to

brown, smooth, dextrinoid, thick-walled, wall <0.7 um thick, non-gelatinous.

Marasmius midnapurensis sp. nov. (India) ... 121

FIGuRE 2. Marasmius midnapurensis (CUH AMT002). Basidiomata. Scale bar = 10 mm.

STIPE TRAMA hyphae parallel, hyaline, smooth, dextrinoid, thin-walled, non-

gelatinous. OLEIFEROUS HYPHAE present, <6.1 um broad. CAULOCYSTIDIA

composed of two types of cells: a) Siccus-type broom cells with main body

28-29.4 x 3.5-4.3 um, scattered, uncommon, irregular in outline, hyaline,

apical setulae 6-6.4 x 1.8-2.1 um, conical to wavy, pale yellow, thin- to thick-

walled, b) non-setulose cells (11—)28.6-42.9(-50.1) x (3.5-)3.9-4.7(-6.3)

um, abundant, cylindrical or irregular in outline, seldom branched, obtuse

to subacute, hyaline, inamyloid, thin- to thick-walled. Clamp CONNECTIONS

present in all tissues.

Molecular & phylogenetic analysis

The amplified fragment of M. midnapurensis with the combination of

primer set ITS1-F (forward) and ITS2 (reverse) produced 259 bp long stretches

including the 226 bp ITS1 region. BLAST analyses with our M. midnapurensis

sequences recovered Marasmius sequences, with the highest similarity shown

by M. jasminodorus Wannathes et al. (92%), M. araucariae var. siccipes

Desjardin et al. (89%), M. aurantioferrugineus Hongo (87%), M. cf. cladophyllus

Berk. (87%), and M. purpureostriatus Hongo (84%).

Phylogenetic analysis of the ITS1 region inferred by the neighbor-joining

method strongly supports M. midnapurensis as a distinct species within

Marasmius and clusters M. midnapurensis and the seven other in-group

122 ... Dutta & al.

Marasmius aurantioferrugineus (FJ904962)

Marasmius aurantioferrugineus (F J904963)

Marasmius aurantioferrugineus (FJ904964)

Marasmius cf. cladophyllus (AY916705)

Marasmius midnapurensis (KF682470)

Marasmius jasminodorous (EU935512)

Marasmius araucariae var. siccipes (F J43 1223)

100 — Marasmius araucariae var. siccipes (F J431222)

Marasmius oreades (DQ490641)

Marasmius wynneae (FJ904979)

Marasmius purpureostriatus (EU935539)

Marasmius rotula (JN714927)

Mycena pura (JN182202)

outgroup

005

FicurE 3. Phylogram showing relationship of Marasmius midnapurensis to closely related species

of Marasmius based on internal transcribed spacer-1 (ITS1) sequences of rDNA. Marasmius

midnapurensis is placed in bold font to highlight its phylogenetic position in the tree.

Marasmius spp. in a clade with 99% bootstrap support. Although

M. midnapurensis forms a distinct branch sister to M. jasminodorus with 93%

bootstrap support in the present study, another closely related species (M.

araucariae var. siccipes, also in sect. Sicci ser. Atrorubentes) forms a separate

clade with 65 % bootstrap support (Fic. 3).

Discussion

The absence of collarium and the presence of Siccus-type broom cells in the

pileipellis, a well developed long central stipe, and adnexed lamellae suggest

that M. midnapurensis belongs to Marasmius sect. Sicci Singer, while the

presence of Siccus-type of broom cells combined with non-setulose cells on the

stipe surface place it in the series Atrorubentes (Wannathes et al. 2009), where it

appears to be closely related to M. araucariae var. siccipes described from Java,

which differs in a more brownish orange pileus and darker lamellae (brownish

orange to greyish brown), and slightly smaller basidiospores (8-12 x 3.5-4 um;

Desjardin et al. 2000).

Basidiospore shape and size, cheilocystidial and caulocystidial characters,

and the absence of pleurocystidia are characters shared with M. jasminodorus

from northern Thailand [ellipsoid basidiospores 9-14 x 3-4.5 um with Q_ of

2.6-3.4, Siccus-type cheilocystidia (9-26 x 6-10 um), caulocystidia of Siccus-

type broom cells and cylindrical non-setulose cells, absence of pleurocystidia].

Marasmius jasminodorus differs macroscopically in its light brown to brownish

pileus, pale yellowish white lamellae with pale brownish orange edges, a fragrant

jasmine tea-like odor, and stipe base with strigose, brownish orange mycelium

(Wannathes et al. 2009). Another similar species, M. koreanus Antonin et al.

N23

Marasmius midnapurensis sp. nov. (India) ...

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124 ... Dutta & al.

from Republic of Korea, differs by brownish orange to reddish orange pileus

with a distinctly brownish orange, rugulose centre and absence of caulocystidia

(Antonin et al. 2012). All other species of Marasmius sect. Sicci described by

Deng et al. (2012) from China differ from M. midnapurensis by the presence

of setae on the pileus and only a single type of caulocystidia. In addition to

the morphological differences (TABLE 1), the ITS sequence analysis clearly

separates M. midnapurensis from M. jasminodorus, M. aurantioferrugineus,

M. cf. cladophyllus, and M. purpureostriatus. Our findings support Tan et al.

(2009) and Wannathes et al. (2009), who found that molecular data do not

support morphologically delimited concepts within Marasmius groups.

The objective of our study was to confirm the taxonomical position of M.

midnapurensis (sect. Sicci ser. Atrorubentes) using ITS rDNA sequence data.

Acknowledgments

The authors are grateful to Armin MeSsi¢ (Ruder BoSskovic Institute, Zagreb,

Croatia), T.K. Arun Kumar (Zamorin’s Guruvayurapan College, Kerala, India), and

Alfredo Justo (Clark University, Worcester, United States) for their critical reviews of

the manuscript. Thanks are also owed to Vladimir Antonin (Brno, Czech Republic)

for his valuable suggestions on the allied species of the newly described taxa and

Patinjareveettil Manimohan (University of Calicut, Malappuram, India) for his guidance

during manuscript preparation. This paper appears through financial support by the

Department of Environment, Government of West Bengal, India.

Literature cited

Anonymous. 1969. Flora of British Fungi. Colour identification chart. Her Majesty’s Stationery

Office, Edinburgh.

Antonin V, Ryoo R, Shin HD. 2012. Marasmioid and gymnopoid fungi of the Republic of Korea.

4, Marasmius sect. Sicci. Mycol. Prog. 11(3): 615-638.

http://link.springer.com/article/10.1007/s11557-011-0773-y.

Berkeley MJ. 1851. Decades of fungi. Decades XXXII, XXXII. Sikkim-Himalayan fungi collected

by Dr. Hooker. Hooker's J. Bot. Kew Gard. Misc. 3: 39-49.

Bilgrami KS, Jamaluddin S, Rizwi MA. 1991. Fungi of India. Today and Tomorrow’s Printers &

Publishers, New Delhi.

Bose SR. 1949. Horse hair fungus from Bengal. Proc. Indian. Sci. Congr., Sec. VI, 36: 123.

Bose SR, Chatterjee P. 1950. A case of apparent symbiosis of Lagerstroemia flos-reginae Retz with

Marasmius campanella Holt. Proc. Indian. Sci. Congr., Sec. 3: 37-56.

Deng C, Li T, Li T, Antonin V. 2012. New species and new records in Marasmius sect. Sicci from

China. Cryptogamie Mycol. 33(4): 439-451. http://dx.doi.org/10.7872/crym.v33.iss4.2012.439

Desjardin DE, Retnowati A, Horak E. 2000. Agaricales of Indonesia: 2. A preliminary monograph

of Marasmius from Java and Bali. Sydowia 52: 92-193.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution

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

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

Marasmius midnapurensis sp. nov. (India) ... 125

Jukes TH, Cantor CR. 1969. Evolution of protein molecules. 21-132, in: HN Munro (ed.).

Mammalian Protein Metabolism. Academic Press, New York.

Manimohan P, Leelavathy KM. 1989. Marasmius species new to India. Sydowia 41: 185-199.

Manjula B. 1983. A revised list of the agaricoid and boletoid basidiomycetes from India and Nepal.

Proc. Indian Acad. Sci., Sect. B, 92: 81-213.

Massee G. 1910. Fungi exotici. X. Bull. Misc. Inf., R. Bot. Gard., Kew 1910: 1-6.

Pradhan P, Banerjee S, Roy A, Acharya K. 2011. Two new species of Marasmius: addition to the

macrofungi of West Bengal, India. Environ. Ecol. 29: 768-770.

Ray S, Samajpati N. 1979. Agaricales of West Bengal IV. Indian J. Mycol. Res. 17: 65-69.

Roy SK. 1953. Notes on some hyphomycetes new to India. Sci. Cult. 19: 94-96.

Saitou N, Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic

trees. Mol. Biol. Evol. 4: 406-425.

Singer R. 1958. Studies toward a monograph of the South American species of Marasmius. Sydowia

12: 54-145.

Singer R. 1986. The Agaricales in modern taxonomy. Koeltz Scientific Books, Federal Republic of

Germany.

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis

(MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599.

http://dx.doi.org/10.1093/molbev/msm092

Tan YS, Desjardin DE, Perry BA, Vikineswary S, Noorlidah A. 2009. Marasmius sensu stricto in

Peninsular Malaysia. Fungal Divers. 37: 9-100.

Wannathes N, Desjardin DE, Hyde KD, Perry BA, Lumyong S. 2009. A monograph of Marasmius

(Basidiomycota) from Northern Thailand based on morphological and molecular (ITS

sequences) data. Fungal Divers. 37: 209-306.

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, London.

Wilson AW, Desjardin DE. 2005. Phylogenetic relationships in the gymnopoid and marasmioid

fungi (Basidiomycetes, euagarics clade). Mycologia 97(3): 667-679.

http://dx.doi.org/10.3852/mycologia.97.3.667

MY COTAXON

http://dx.doi.org/10.5248/128.127

Volume 128, pp. 127-130 April-June 2014

A new species of Arachnophora from submerged wood

in the Amazon rainforest, Brazil

JOSIANE SANTANA MONTEIRO’, LUIS FERNANDO PASCHOLATI GUSMAO”",

& RAFAEL E CASTANEDA-RUIZ?

‘Departamento de Micologia, Universidade Federal de Pernambuco, 50670-420, Recife, Brazil

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

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

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

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

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

* CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT — Arachnophora combuensis sp. nov. is described and illustrated from specimens

collected on submerged wood from Combu Island (Belém municipality, Para, Brazil) in

the Amazon rainforest. The fungus is characterized by simple pale brown conidiophores,

monoblastic percurrently extending conidiogenous cells, and staurosporous conidia with a

pale brown basal cell and 2-3 dark brown central cells that bear 1-3 conical pale brown arms.

The new species is compared with morphologically similar taxa.

KEY worDs — taxonomy, freshwater fungi, tropics

Introduction

During a mycological survey of conidial fungi occurring on submerged

decaying plant materials from the Amazon rainforest in Brazil, an interesting

fungus was collected on submerged wood. Its conidiogenesis and conidial

features clearly suggest placement within the genus Arachnophora Hennebert

(Hennebert 1963). However, the fungus shows remarkable differences from all

previously described Arachnophora species and therefore is described as new

to science.

Material & methods

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

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

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

128 ... Monteiro, Gusmao, & Castafeda-Ruiz

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

bright field and Nomarski interference optics. The type specimens are deposited in the

Herbarium of Universidade Estadual de Feira de Santana (HUEFS).

Taxonomy

Arachnophora combuensis J.S. Monteiro, R.F. Castafieda & Gusmao, sp. nov. Fic. 1

MycoBank MB 807635

Differs from Arachnophora crassa by its wider conidial central body and fewer, larger

conidial arms and from A. pulneyensis by its wider conidial central body separated from

the conidiogenous cell by a basal cell.

Type: Brazil, Para State, Belém, Area de Protecdo Ambiental Ilha do Combu, 1°25’S

48°27’ W, on submerged wood, 10 Jan. 2013, coll. J.S. Monteiro (Holotype: HUEFS

196432).

ErymMoLoecy: Latin combuensis refers to the island where the type specimen was

collected.

Cotonigs on the natural substrate effuse, hairy, dark brown. Mycelium

superficial and immersed. Hyphae branched, septate, smooth, pale brown,

2-4 um wide. CONIDIOPHORES macronematous, mononematous, simple or

with one branch, erect, straight or slightly flexuous, cylindrical, 3-4-septate,

smooth, pale brown, 87.5-225 x 5-7.5 um; branches cylindrical, 1-2-septate,

smooth, pale brown, 27.5-60 x 6-7.5 um. CONIDIOGENOUS CELLS monoblastic,

integrated, terminal, cylindrical, with enteroblastic percurrent extensions,

smooth, pale brown. Conidial secession rhexolytic. ConrbIA acrogenous,

solitary, staurosporous, 42-50(-60) x 13-16 um, composed of a basal cell that

is conical, smooth, pale brown, 7-14 x 5-11 um, with a tapering base bearing

remains of the conidiogenous cells, 2-3 um wide; 1-2(-3) central cells that are

dark brown, smooth, 7-21 x 9-16 um; 1-3(-4) arms that are conical, 1-septate,

12-30 x 5-11 um, with a pale brown basal cell, 6-15 x 5-11 um and a pale

brown distal cell, 7-19 x 3-6 um. Synanamorph Selenosporella-like, present

at the tip of conidial arms, producing conidia sympodially, aseptate, fusiform,

straight, hyaline, 3-5 x 0.5-1 um.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, ParA, Area de Protecdo Ambiental Ilha

do Combu, 1°25’S 48°27’W, on submerged wood, 10 Jan. 2013, coll. J.S. Monteiro

(HUEES 196433).

Note: Arachnophora is typified by A. fagicola Hennebert, which was collected

from decaying cupules of Fagus sylvatica in Belgium (Hennebert 1963).

The genus comprises nine species characterized by monoblastic integrated

percurrently extending conidiogenous cells that produce staurosporous conidia

with dark central cells and paler septate incurved arms (Révay & G6éncz6l 1989,

Castafieda-Ruiz et al. 1996, 1997, Castafteda-Ruiz & Guarro 1998, Pratibha

et al. 2011, Leao-Ferreira et al. 2013, Ma et al. 2013). Among the described

Arachnophora combuensis sp. nov. (Brazil) ... 129

N ‘ie Pp

Fic. 1. Arachnophora combuensis (holotype, HUEFS 196432). A-B. Conidiophores.

C. Conidiogenous cell attached to a conidium. D-P. Conidia. Scale bars: A-B = 30 um; C-P = 20 um.

species only Arachnophora crassa Révay & J. Gonczdl and A. pulneyensis

(Subram. & Bhat) R.E. Castafieda are morphologically similar to A. combuensis.

Arachnophora crassa is distinguished by conidia with 5-7 arms measuring

9-14.5 x 5-6.5 um and a two-celled central body of 16-22.5 x 8-9.5 um (Révay

& Goncz6l 1989). Arachnophora pulneyensis differs from A. combuensis by the

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

sizes of the central body (16-24 x 6.5-8 um) and conidial arms (14 x 3-5 um)

and the absence of a basal cell (Castaneda-Ruiz et al. 1996).

Acknowledgments

The authors express their sincere gratitude to Dr. De- Wei Li and Dr. Xiu-Guo Zhang

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

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

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

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

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

P131LH003033, Cuban Ministry of Science, Technology and Environment for facilities.

Literature cited

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

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

Castanieda-Ruiz RF, Guarro J. 1998. Two new hyphomycetes from rainforest of Cuba. Can. J. Bot.

76: 1584-1588. http://dx.doi.org/10.1139/b98-095

Castafieda-Ruiz RF, Guarro J, Cano J. 1996. Notes on conidial fungi. X. A new species of

Ceratosporella and some new combinations. Mycotaxon 60: 275-281.

Castafieda-Ruiz RF, Gams W, Saikawa M. 1997. Three new conidial fungi (hyphomycetes) from

Cuba. Nova Hedwigia 64: 473-483.

Hennebert GL. 1963. Un hyphomycéte nouveau Arachnophora fagicola gen. nov. spec. nov. Can. J.

Bot. 41(8): 1165-1169. http://dx.doi.org/10.1139/b63-097

Leao-Ferreira SM, Gusmao LFP, Castafieda-Ruiz RE 2013. Conidial fungi from the semi-arid

Caatinga biome of Brazil. Three new species and new records. Nova Hedwigia 96: 479-494.

http://dx.doi.org/10.1127/0029-5035/2013/0084

Ma J, Xia JW, Zhang XG, Castafieda-Ruiz RE 2013 [“2014”]. Arachnophora dinghuensis

sp. nov. and Websteromyces inaequale sp. nov., and two new records of anamorphic

fungi from dead branches of broad-leaved trees in China. Mycoscience 55(5): 329-335.

http://dx.doi.org/10.1016/j.myc.2013.11.007

Pratibha J, Bhat S$, Raghukumar. 2011. Four anamorphic fungi (with two new species) from forests

of Western Ghats, India. Mycotaxon 117: 269-278. http://dx.doi.org/10.5248/117.269

Révay A, Génczél J. 1989. Some dematiaceous hyphomycetes from woody-litter in Hungary. Nova

Hedwigia 48: 237-245.

MY COTAXON

http://dx.doi.org/10.5248/128.131

Volume 128, pp. 131-135 April-June 2014

Xiuguozhangia, a new genus of microfungi to accommodate

five Piricaudiopsis species

Kat ZHANG’, L1-Guo Ma’, JIAN Ma’, & RAFAEL F. CASTANEDA-RUvIz3

"Department of Landscaping, Shandong Yingcai University,

Jinan, Shandong, 250104, China

?Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong, 271018, China

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

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

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

*CORRESPONDENCE TO: kaise0907@126.com

ABSTRACT — Xiuguozhangia gen. nov. is established to accommodate five species described

in Piricaudiopsis but characterized by holoblastic conidial ontogeny. Five new combinations

are proposed: Xiuguozhangia appendiculata, X. indica, X. punicae, X. rhaphidophorae, and

X. rosae. Notes and illustrations on the type species of Piricaudiopsis, P. elegans, are also

provided.

KEY worpDs — asexual fungi, systematics, leaf litter

Piricaudiopsis J. Mena & Mercado, typified by P elegans J. Mena & Mercado

(Mena Portales & Mercado Sierra 1987), is characterized by conidiophores that

are macronematous, mononematous, unbranched, erect, cylindrical, and dark

brown and conidiogenous cells that are monotretic, integrated, terminal and

discrete, intercalary, doliiform, and subspherical to rather lageniform (Fie. 1).

Conidial ontogeny is enteroblastic; the conidia are dictyoseptate, with several

compact rows of cells, somewhat complanate, brown, verrucose towards the

base, verruculose or smooth towards the distal cells; 4-7 of the apical cells

produce a divergent, straight to slightly curved arm.

Five additional species have been described in Piricaudiopsis: P appendiculata

(Bhat & Kendrick 1993); P. indica (Sureshkumar et al. 2005); and P punicae,

P. rhaphidophorae, and P. rosae (Zhang et al. 2009). All five of these species

have holoblastic conidial ontogeny (in contrast to the enteroblastic conidial

132 ... Zhang & al.

ontogeny of P elegans). We therefore propose a new genus, to which we transfer

the five holoblastic species.

Taxonomy

Fic 1. Piricaudiopsis elegans (INIFAT C06/70). A. Conidia. B. Conidiogenous cells.

Fic 2. Xiuguozhangia rosae (HSAUPVII, .... 1092). Conidiogenous cells and conidia.

0-KAI

Xiuguozhangia K. Zhang, R.F. Castafieda, Jian Ma & L.G. Ma, gen. nov.

MycoBank MB 807641

Differs from Piricaudiopsis by its holoblastic conidial ontogeny.

TYPE SPECIES: Xiuguozhangia rosae (K. Zhang & X.G. Zhang) K. Zhang & R. F. Castaneda

ErymoLoey: Latin Xiuguozhangia, dedicated to Prof. Xiu Guo Zhang (Shandong

Agricultural University, China) for his contribution to microfungal taxonomy.

CoLonigs on the natural substrate effuse, hairy, velutinous, brown to dark

brown. Mycelium mostly superficial immersed. ConrpiopHoREs distinct,

single, cylindrical, erect, straight or flexuous, unbranched or moderately

branched, multiseptate, brown to dark brown below, mid to pale brown toward

Xiuguozhangia gen. nov. ... 133

Fig 3. Xiuguozhangia appendiculata (DAOM 214610). Conidiogenous cell and conidium.

Fic 4. Xiuguozhangia indica (ex HCIO-45498). Conidiogenous cell and conidium. Redrawn from

original publication.

the apex, smooth or verruculose. CONIDIOGENOUS CELLS monoblastic, mostly

discrete, few, terminal, integrated, lageniform, determinate, or indeterminate

with several enteroblastic percurrent elongations, pale brown. Conidiogenous

loci flat, truncated. CONIDIAL ONTOGENY holoblastic. CONIDIAL SECESSION

schizolytic. Conrp1A solitary, acropleurogenous, dictyoseptate, campanulate,

cheiroid, composed of several compact rows of cells, that form a compact

cluster after successive + dichotomous branchings, brown, verrucose towards

the base, verruculose or smooth towards the distal cells; 0-4 of the apical cells

may produce a slender, divergent, straight to slightly curved, septate, pale

brown, appendage.

Xiuguozhangia rosae (K. Zhang & X.G. Zhang) K. Zhang & R.E. Castafieda,

comb. nov. FIG. 2

MycoBank MB 807642

= Piricaudiopsis rosae K. Zhang & X.G. Zhang, Mycologia 101(3): 419 (2009).

Xiuguozhangia appendiculata (Bhat & W.B. Kendr.) K. Zhang & R.E. Castafieda,

comb. nov. FIG. 3

MycoBank MB 807643

= Piricaudiopsis appendiculata Bhat & W.B. Kendr., Mycotaxon 49: 62 (1993).

134 ... Zhang & al.

0432)Conidiophore, conidiogenous cells and

1417). A. Conidia. B—C. Conidio-

Fic 5. Xiuguozhangia punicae (HSAUPVII

0-KAI

conidia. Fic 6. Xiuguozhangia rhaphidophorae (HSAUPVII

phores, conidiogenous cells and conidia.

0-KAI

Xiuguozhangia indica (Sharath, Sureshk., Kunwar & Manohar.) K. Zhang & R.E.

Castafieda, comb. nov. FIG. 4

MycoBank MB 807644

= Piricaudiopsis indica Sharath, Sureshk., Kunwar &

Manohar., Mycotaxon 92: 280 (2005).

Xiuguozhangia punicae (K. Zhang & X.G. Zhang) K. Zhang & R.F. Castafieda,

comb. nov. Fig: §

MycoBank MB 807646

= Piricaudiopsis punicae K. Zhang & X.G. Zhang, Mycologia 101(3): 417 (2009).

Xiuguozhangia rhaphidophorae (K. Zhang & X.G. Zhang) K. Zhang & RE.

Castaneda, comb. nov. FIG. 6

MycoBAnk MB 807645

= Piricaudiopsis rhaphidophorae K. Zhang & X.G. Zhang, Mycologia 101(3): 420 (2009).

Xiuguozhangia gen. nov. ... 135

Acknowledgments

The authors express their sincere gratitude to Dr. Luis F. P. Gusmao and Dr. De-Wei Li

for their critical review of the manuscript. This project was supported by the National

Natural Science Foundation of China (Nos. 31200013, 31093440, 30770015, 30499340)

and the Ministry of Science and Technology of the People’s Republic of China (Nos.

2006FY120100, 2006FY110500-5). We also thank to Cuban Ministry of Agriculture

(MINAG) and CITMA (Project P131LH003033, Program Salud animal y Vegetal)

for facilities. Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun Pennycook’s

nomenclature review are greatly appreciated.

Literature cited

Bhat DJ, Kendrick B. 1993. Twenty-five new conidial fungi from the Western Ghats and the

Andaman Islands (India). Mycotaxon 49: 19-90.

Mena Portales J, Mercado Sierra A. 1987. Piricaudiopsis (Hyphomycetes, Deuteromycotina), nuevo

genero enteroblastico de Cuba. Acta Botanica Cubana. 51: 1-5.

Sureshkumar G, Sharath Babu K, Kunwar IK, Manoharachary C. 2005. Two new hyphomycetous

fungal species from India. Mycotaxon 92: 279-283.

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

Mycologia 101: 417-422. http://dx.doi.org/10.3852/08-147

MY COTAXON

http://dx.doi.org/10.5248/128.137

Volume 128, pp. 137-144 April-June 2014

Hymenochaete in China. 8. H. biformisetosa sp. nov.

with a key to species with denticulate setae

JIAO YANG & SHUANG-Hut1 HE*

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

CORRESPONDENCE TO*: shuanghuihe@yahoo.com

AxBsTRACT — Hymenochaete biformisetosa, a new species from Yunnan Province in

southwestern China, is described and illustrated. It is characterized by thick, resupinate

basidiocarps, two types of setae (sometimes denticulate), and ellipsoid to broadly ellipsoid

basidiospores. The new species is similar to H. spinulosetosa and H. denticulata but with

morphological and molecular (ITS rDNA sequences) differences. An identification key to

nine species of Hymenochaete with denticulate setae is provided.

Key worps — Hymenochaetaceae, phylogeny, taxonomy, wood-inhabiting fungi

Introduction

The diversity of wood-inhabiting fungi in Yunnan Province, southwest

China, is extremely high because of abundant vascular plants and complex

landforms. Although many basidiomycete species have been described from

this area in recent years (Dai et al. 2002; Yuan & Dai 2008a,b; Dai & Li 2010,

Dai 2011, 2012; He & Li 2011, 2012, He & Dai 2012; Zhao et al. 2013), corticioid

fungi are under-collected and studied.

In 2012, an intensive investigation of wood-inhabiting fungi was carried out

in the Gaoligong Mountains in northwest Yunnan Province, which is considered

an important biodiversity hotspot in China. Among the approximately 100

specimens collected in this survey, an undescribed species of Hymenochaete

was found. We describe and illustrate the new fungus as H. biformisetosa in our

new contribution to the knowledge of Hymenochaete species in China.

Materials & methods

MORPHOLOGICAL STUDIES: Voucher specimens were deposited in the herbarium of

Beijing Forestry University (BJFC), and the microscopic procedure followed Dai (2010).

138 ... Yang & He

H. acerosa JQ279543

98 [L_ FF cinnamomea JQ279548

T\ L Ff minuscula 1Q279546

H. epichlora JQ279549

H. unicolor JQ279551

100 -— H. minor JQ279555

4 H. parmastoi JQ780061

H. megaspora JQ279553

86;— H. ochromarginata JQ279579

100 |" H. rubiginosa JQ716407

H. tasmanica JQ279582

54- H. paucisetigera JQ279560

100L_ 7 asetosa JQ279559

H. odontoides JQ279563

H. fulva JQ279565

H. yunnanensis JQ279571

93 H. anomala JQ279566

H. luteobadia JQ279569

H. separabilis JQ279572

68 H. rhabarbarina JQ279574

H. ustulata JQ780066

93 H. huangshanensis JQ279533

H. longispora JQ279536

H. ulmicola JQ780065

H. tongbiguanensis KF908248*

H. rhododendricola JQ279576

100; H. attenuata JQ279526

H. villosa JQ279528

H. rheicolor JQ279530

91 H. adusta JQ279523

H. subferruginea JQ279521

H. xerantica JQ279519

98 H. berteroi JQ279524

60°- H. setipora JQ279515

97 FF porioides JQ279517

H. cyclolamellata JQ279513

H. nanospora JQ279531

H. muroiana JQ279542

100; H. floridea JQ279597

H. sphaericola JQ279599

= H. cruenta 3Q279595

‘. ae H. biformisetosa KF908247*

H. denticulata KF438171

ne 95 H. legeri JQ780067

H. spathulata JQ279591

36 H. murina JQ716406

100 _- H. duportii DQ404386

H. hydnoides JQ279590

H. sphaerospora JQ279593

100 H. innexa JQ279584

H. tropica JQ279588

A Pseudochaete tabacina JQ279610

Fic. 1. Strict consensus tree obtained from Maximum Parsimony analysis of ITS sequences of

Hymenochaete species. Pseudochaete tabacina was used as outgroup. Newly generated sequences

are indicated by an asterisk (*). Parsimony bootstrap values >50% are shown.

Hymenochaete biformisetosa sp. nov. (China) ... 139

In presenting the size range of spores and setae, 5% of the measurements from each end

of the range are given in parentheses. Special color terms follow Petersen (1996).

DNA EXTRACTION AND SEQUENCING: A CTAB Plant genome rapid extraction kit DN14

(Aidlab Biotechnologies Co., Ltd) was used to extract DNA from dried specimens

following the manufacturer’s instructions. Approximately 700 base pairs of the ITS

rDNA were amplified with primers ITS5 and ITS4 as follows: initial denaturation at 95

°C for 3 min, followed by 38 cycles of 94 °C for 40 s, 58 °C for 45 s and 72 °C for 1 min,

and a final extension of 72 °C for 10 min. DNA sequencing was performed at Beijing

Genomics Institute, and all the newly generated sequences were deposited in GenBank.

PHYLOGENETIC ANALYSIS: The phylogeny was inferred from the ITS sequences of 51

Hymenochaete species. Pseudochaete tabacina (Sowerby) T. Wagner & M. Fisch. was

selected as outgroup. The sequences were aligned using the ClustalX 1.83 (Chenna et al.

2003). Alignments were optimized manually in BioEdit 7.0.5.3 (Hall 1999). Maximum

parsimony analysis were performed using PAUP* 4.0b10 (Swofford 2002). Gaps in the

alignments were treated as missing data. Trees were generated using 100 replicates of

random stepwise addition of sequence and tree-bisection reconnection (TBR) branch-

swapping algorithm, with all characters given equal weight. Branch supports for all

parsimony analysis were estimated by performing 1000 bootstrap replicates (Felsenstein

1985) with a heuristic search of 10 random-addition replicates for each bootstrap

replicate. The tree length (TL), consistency indices (CI), retention indices (RI), rescaled

consistency indices (RC) and homoplasy index (HI) were calculated for each tree

generated. Trees were figured in Treeview 1.6.6 (Page 1996).

Phylogenetic results

Two new ITS sequences were obtained in this study (GenBank accession

numbers KF908247 and KF908248). The alignment of the ITS sequences of

52 taxa resulted in 764 sites with 389 parsimony informative characters. Two

equally parsimonious trees were obtained from the analysis with TL = 2180,

CI = 0.423, RI = 0.588, RC = 0.249 and HI = 0.577. The strict consensus tree

showed that Hymenochaete biformisetosa formed a distinct lineage embedded

in a clade with H. denticulata J.C. Léger & Lanq., H. cruenta (Pers.) Donk,

H. floridea Berk. & Broome, and H. sphaericola Lloyd (Fie. 1).

Taxonomy

Hymenochaete biformisetosa Jiao Yang & S.H. He, sp. nov. Figs 2-4, 8

MycoBank MB 807093

Differs from other Hymenochaete species by its thick basidiocarps, two types of setae

that may be denticulate, and ellipsoid to broadly ellipsoid basidiospores.

Type: China. Yunnan Prov., Baoshan, Gaoligongshan Nat. Res., Baihualing, on

angiosperm stump, 28.X.2012, He 1445 (holotype, BJFC 14297; GenBank KF908247).

Erymo_oey: the epithet “biformisetosa” refers to the two types of setae.

FRuITBODy: Basidiocarps annual, effused, closely adnate, corky to woody

hard, first as small round or irregular patches, later confluent up to 6 x 2 cm,

140 ... Yang & He

—_ “

Fics. 2-3. Hymenochaete biformisetosa (holotype): 2. Basidiocarps. 3. Close-up of hymenophore

surface. 4. Wood decayed by H. biformisetosa. Fics. 5-7. Basidiocarps of other Hymenochaete

species with denticulate setae: 5. H. denticulata (He 1271). 6. H. separabilis (He 271).

7. H. tongbiguanensis (He 1552).

0.15-2.5 mm thick in section. Hymenophore smooth, grayish brown to fawn,

not cracked when young, becoming pale mouse-gray to light vinaceous gray,

deeply and sparsely cracked when mature; margin thinning out, reddish brown

to orange-brown, distinct, up to 1 mm wide when juvenile, becoming fuscous

or concolorous with hymenophore surface, narrower with age.

HyPHAL STRUCTURE: Hyphal system subdimitic; generative hyphae without

clamp connections; tissue darkening but otherwise unchanged in potassium

hydroxide.

SUBICULUM: Tomentum and cortex absent. Hyphal layer absent or

sometimes present, consist of interwoven hyphae. Generative hyphae rare,

Hymenochaete biformisetosa sp. nov. (China) ... 141

7 Tm

——

10 pm

—

40pm

Fic. 8. Hymenochaete biformisetosa (holotype):

a. Basidiospores. b. Basidia and basidioles. c. Small setae. d. Large setae.

hyaline to yellowish brown, thin- to slightly thick-walled with a wide lumen,

septate, moderately branched, 2-4 um in diam. Skeletoid hyphae predominant,

reddish brown, distinctly thick-walled with a narrow lumen, some parts nearly

solid, usually developing into setal hyphae with acute apices, 3-4.5 um in diam.

STRATIFIED HYMENIUM: Hyphae similar to those in subiculum, reddish

brown, thick-walled, agglutinated, interwoven. Setal layer thick, composed

of several to many rows of overlapping setae. Setae of two types (1) large,

numerous, subulate, reddish brown, with acute tips, projecting up to 90 um

above the hymenium, (70—)80-130(-140) x (8-)9-14(-16) um, often with

crystals loosely attached to the projected part when mature; (2) small, numerous,

subulate, reddish brown, with acute tips, some with a hyphal tail, usually not

protruding, (30-)35-50(-60) x 4-8 um. Both types of setae sometimes with

1-3 conical spines near apex; spines up to 2 um long. Cystidia and hyphidia

absent. Basidia subcylindrical to subclavate, usually restricted in central part,

with four sterigmata and a simple septum at base, 20-25 x 3-5 um; basidioles

similar to basidia in shape but shorter.

142 ... Yang & He

Spores: Basidiospores ellipsoid to broadly ellipsoid, hyaline, thin-walled,

smooth, sometimes containing one guttule, inamyloid and nondextrinoid in

Melzer’s reagent, acyanophilous in cotton blue, (4.1-)4.3-6(-6.5) x (2.8-)3-4.2

(-4.5) um, mean spore length = 5.2 um, mean spore width = 3.5 um, length/

width = 1.5 (60 spores of one specimen were measured).

TYPE OF ROT: Causes a white pocket rot of angiosperm wood.

ADDITIONAL SPECIMENS EXAMINED: Hymenochaete spinulosetosa. VENEZUELA.

MERIDA Prov., Monte Zerpa, on dead hardwood, 29.1.2001, Ryvarden 43707 (isotype,

TAAM 202926).

Hymenochaete denticulata. CHINA. GUANGxI AUTONOMOUS REGION, Longzhou

County, Nonggang Nat. Res., on fallen angiosperm twig, 21.VII.2012, He 1271 (BJFC

14122; Fic. 5).

Hymenochaete separabilis: CHINA. ANHUI PRov., Huangshan County, Huangshan Nat.

Res., on fallen angiosperm trunk, 21.X.2010, He 460 (BJFC 9938; Fic. 6).

Hymenochaete tongbiguanensis: CHINA. YUNNAN PROV., Ruili, on fallen angiosperm

branch (paratype, HMAS 84867); Longchuan County, Tongbiguan Nat. Res., on

angiosperm stump, 31.X.2012, He 1497 (BJFC 14352). HaInan Prov., Lingshui

County, Diaoluoshan Nat. Res., on angiosperm stump, 10.X1.2012, He 1552 (BJFC

14407; GenBank KF908248; Fic. 7).

Discussion

Hymenochaete biformisetosa can be easily distinguished from other species

of the genus by its thick (<2.5 mm) resupinate basidiocarps, two types of setae

that sometimes are denticulate, and ellipsoid to broadly ellipsoid basidiospores;

it causes a distinct white pocket rot of angiosperm wood.

Two species, Hymenochaete tongbiguanensis T.X. Zhou & L.Z. Zhao and

H. spinulosetosa Parmasto, also have thick basidiocarps and denticulate setae.

Hymenochaete tongbiguanensis can be distinguished from H. biformisetosa by its

smaller setae (22.5-42 x 3.5-8 um) and basidiospores (3-4 x 1.5-2 um; Zhang

& Dai 2005), and H. spinulosetosa has large, broadly ellipsoid basidiospores

(5-6.2 x 3.2-3.7 um) but only one type of setae (50-75 x 5-7.5 um; Parmasto

20122

Six other species of Hymenochaete, including H. denticulata, also produce

denticulate setae, but they differ from H. biformisetosa in having thin

basidiocarps (<150 um), shorter setae (40-60 um), and shorter basidiospores

(<5 um, Léger 1998; Parmasto 2012). Hymenochaete unicolor Berk. & M.A.

Curtis is similar to H. biformisetosa in basidiocarp morphology and anatomical

structure with smaller, smooth setae (35-60 x 5-9 um, Parmasto 2005).

In the phylogenetic tree (Fic. 1), Hymenochaete biformisetosa appears to be

closely related to H. denticulata (bootstrap value = 92%), which also has two

types of denticulate setae (Léger 1998; He & Li 2014). The differences between

these two species are discussed above. The new species is related also to the

Hymenochaete biformisetosa sp. nov. (China) ... 143

H. cruenta group in the phylogenetic tree (Fic. 1); however, species of this group

have red-colored basidiocarps, dendrohyphidia, and cylindrical basidiospores

(Léger 1998; Parmasto 2001; Parmasto & Gilbertson 2005).

Of the nine Hymenochaete species with denticulate setae now described

(Gilbertson & Hemmes 1997; Léger 1998; Zhang & Dai 2005; Parmasto 2012),

four are known from China (Zhang & Dai 2005; He & Li 2014).

Key to species of Hymenochaete with denticulate setae

1} Basidiocarps-<0515 min thick: 2. ies ose x5 eas see vee Meat eas es BS eS Sis 2

it, Basidiocarps up to clear tOF Tae eA 2d Uk Ue el Ae Ad RL 7

2. Well-developed'hyphallayer presente... 9.0... 908 2 98 PE ae OE ot ee ced art aa 3

ZU Nab AV CADSEIS 2s tac sca tte Harkenn Haen thy Sawhiee AMAR RMR Ees vases qokwer Pana bases 4

Sesetae 30-40: umblong,witheld teeth, 0, ust, wines Eels » encles seinen H. separabilis

3. Setae 40-70 um long, with 5-10 teeth..................0.0.00.. H. tomentelloidea

4, Setad with 1a) teeth, 5 eg ov sore oes Bee ov ag a aes ale als sole ale aS als ae H. pellicula

#Setae awith-several fo winaniy CCU Far ii Pge antl Page ae age Page OR ag le Bese oP lo BE aa 5

SHSCLACIOL LWVONLV Ie Sa Sy tha Se ctegealsise nea pate eae pen geen goes oc Bee H. denticulata

SPT E:( AU LIVECE) G20 Ry CULE Ree Oe mE SOLE aN LAE SAE JOM RON Ont Sane Gre oe OO xe ERNE SON 4 6

6. Cortex present; spores subglobose, 3.7—4.3 um wide.................0. H. harpago

6. Cortex absent; spores ellipsoid to cylindrical, 1.5-2 um wide.......... H. pertenuis

7. Setae <50 um long; spores 3-4 x 1.5-2 um................005. H. tongbiguanensis

Fesetae 250 pin lONne: Spores PLOTS MIM... teen eee eee ees ek a ee eee oe 8

8. Setae of two types, large setae up to 130 um long................. H. biformisetosa

SiSelae UNM, Wp 1975 WON, ners becncitin Mionatees Meenas Menetey Benet H. spinulosetosa

Acknowledgments

The authors would like to express their deep thanks to Drs. Karen Nakasone

(Northern Research Station, U.S. Forest Service, USA) and Hai-Sheng Yuan (Institute of

Applied Ecology, Chinese Academy of Sciences, China) for serving as pre-submission

reviewers, to Prof. Yu-Cheng Dai (Beijing Forestry University, China) for improving

the manuscript, to Dr. Hai-Jiao Li for drawing the microscopic illustration. This study

was supported by the Fundamental Research Funds for the Central Universities (No.

YX2013-04).

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MY COTAXON

http://dx.doi.org/10.5248/128.145

Volume 128, pp. 145-157 April-June 2014

New records of Dothideomycetes from Mexico

SANTIAGO CHACON’ , FIDEL TAPIA!, & MART{iN ESQUEDA?

‘Instituto de Ecologia, A.C. Apartado Postal 63, Xalapa, Veracruz 91000, México

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

Apartado Postal 1735, Hermosillo, Sonora 83304, México

* CORRESPONDENCE TO: santiago.chacon@inecol.edu.mx

Asstract —Ten dothideomycetous species were studied. New records for the Mexican

mycobiota include Heptameria obesa, Leptospora rubella, Macrovalsaria megalospora, and

Psiloglonium clavisporum, and an extended distribution for rarely reported dothideomycetes

in Mexico is reported for Anteaglonium abbreviatum, Astrosphaeriella trochus, Gloniopsis

praelonga, Hysterobrevium mori, Oedohysterium insidens, and Rhytidhysteron rufulum.

Observations and photographs on macro- and microscopic characters are provided.

KEY worps— Ascomycota, taxonomy, chorology

Introduction

Most fungi with ascostromatic development and bitunicate asci are generally

associated with the Dothideomycetes, the ascomycete class with the highest

number of species (slightly more than 19,000; Kirk et al. 2008). They are found

in nature as pathogens, endophytes or epiphytes of living plants, and saprobes

that degrade cellulose or other carbohydrates in organic matter, litter, and dung.

Knowledge about ascomycetes in Mexico is still low compared to some

European countries where more research has been completed on ascomycete

mycological diversity. In Mexico, Medel (2007a) cited 687 ascomycete species,

while Gonzalez & Hanlin (2008) reported 880 species, excluding lichenized

ascomycetes. Pezizales and Xylariales are represented by the greatest number

of species records. The first record of a dothideomycetous species for Mexico

dates back more than 60 years to Sphaeria pertusa Pers. (= Trematosphaeria

pertusa (Pers.) Fuckel) cited by Fries (1851) in Veracruz. Publications on

dothideomycetous species of Mexico include Hyde (1992), San Martin (1996),

San Martin & Lavin (1999), Checa et al. (2002), Medel (2007b), and Méndez-

Mayboca et al. (2010).

146 ... Chacon, Tapia, & Esqueda

Materials & methods

Specimens of Dothideomycetes examined included those deposited in the XAL

fungal collection as well as collections obtained by S$. Chacon & F. Tapia in Veracruz and

southeast Mexico during November 2011-September 2012.

Hysterothecia and pseudothecia were examined using a stereoscopic microscope,

and microscopic structures were examined using a light microscope. Ascomata were

sectioned using a razor blade and mounted in KOH (3% or 5%) and Meltzer solutions

as needed. Asci, ascospores, pseudoparaphyses, and peridial thickness were measured.

Species were identified consulting Zogg (1962), Dennis (1981), Sivanesan (1984),

Barr (1990a,b), and Boehm et al. (2009). Complete descriptions are provided for species

newly recorded for Mexico, and a taxonomic summary and brief notes are presented

for the species previously recorded. The recent collections have been deposited in the

Herbarium of the Instituto de Ecologia A.C. Xalapa (XAL), Veracruz, Mexico.

Taxonomy

Anteaglonium abbreviatum (Schwein.) Mugambi & Huhndorf, Syst. Biodiv. 7(4):

460 (2009) FIGS. 1, 15

= Glonium abbreviatum (Schwein.) M.L. Lohman

OBSERVATIONS— Anteaglonium abbreviatum is characterized mainly by linear

hysterothecia with truncated apices, which allow some blackening on the

substrate and spores that are fusiform, bicellular, constrained at the septum,

and measure (6.5—)7-—7.5(-8) x 2.5-3 um. Anteaglonium globosum Mugambi

& Huhndorf (6-7 x 2-3 um) and A. parvulum (W.R. Gerard) Mugambi &

Huhndorf (6-8 x 2.5-3 um) have similarly sized and shaped spores (Boehm

et al. 2009) but differ in the form of hysterothecium. In A. globosum the

hysterothecium is globose (as the name suggests) and roughened, while in

A. parvulum the hysterothecium is linear but with acuminated apices. Spore

sizes for A. abbreviatum (as Glonium abbreviatum) have been described as

measuring 6-8 x 2-5 um (Lohman 1937), (5-)6-7(-8) x 2-3(-3.5) um (Zogg 1962),

and 7-8 x 3 um (Romero 1987). Anteaglonium abbreviatum is widely distributed

and was recently cited by Méndez-Mayboca et al. (2010, as G. abbreviatum)

from Sonora. It is reported for the State of Veracruz here for the first time.

HasiTaT— On dead wood in forest relict of Fagus grandifolia var. mexicana

(Martinez) Little, 850-1900 m asl.

SPECIMENS EXAMINED — MEXICO. VerAcRuz: Municipality of Emiliano Zapata,

CHAVARRILLO-EL PALMAR ROAD on Monte Oscuro summit, 13 October 2004, S. Chacon

5530; Municipality of Acatlan, ACATLAN VOLCANO upper south part, 11 November

2011, S. Chacén & F. Tapia 6613; Municipality of Xalapa, PARQUE NATURA natural

protected area, 1 November 2011, S. Chacon & E Tapia 6585; HousE GARDEN on Adolfo

Ruiz Cortines Ave., ~200 m from Hospital de Especialidades Médicas, 27 August 2012,

S. Chacon & F. Tapia 6849; CoLonta EL O_o, ex-hacienda Las Animas, 27 August

2012, S. Chacén & FE. Tapia 6861. SonoRA: Municipality of Alamos, EL Acuayg, 14

September 2006, S. Chacon 5764.

New Dothideomycetes in Mexico ... 147

eive

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Fics 1-6. Anteaglonium abbreviatum, 1: ascospores. Astrosphaeriella trochus, 2: ascospores.

Gloniopsis praelonga, 3: ascospores. Heptameria obesa, 4: mature and immature asci. 5: ascospores.

Hysterobrevium mori, 6: ascospores. Scale bars: 1= 5 um; 2-6= 10 um

Astrosphaeriella trochus (Penz. & Sacc.) D. Hawksw., Bot. J. Linn. Soc. 82: 46 (1981)

FIG. 2, 16

OBSERVATIONS— ‘The studied material is consistent with Hawksworth (1981,

except for the spore color, cited by Hawksworth as reddish-brown), and agrees

with San Martin & Lavin (1999, who described olivaceous-brown to pale-

148 ... Chacon, Tapia, & Esqueda

brown spores). This is the second record of the species in Mexico and the first

for the state of Veracruz. Previously it was recorded for the state of Chiapas by

San Martin & Lavin (1999).

HasiTatT— On stems of Bambusa sp. and an undetermined grass in a semi-

deciduous forest and a coffee plantation; 460-1300 m asl.

SPECIMENS EXAMINED —- MEXICO. Veracruz: Municipality of Actopan, Km 28

XALAPA-VERACRUZ HIGHWAY, near the highest turn to Balastrera, 7 September 1995,

S. Chacén 4930; Municipality of Xico, NEAR TEXOLO WATERFALL, 22 October 1999, S.

Chacon 5216; Municipality of Xalapa, EL OLMO sECTION, southeast Xalapa, 15 August

2012, S. Chacon & E. Tapia 6820; FRANCISCO JAVIER CLAVIJERO BOTANIC GARDEN, Km

2.5 old road Xalapa-Coatepec, 22 August 2012, F. Tapia 2637; INECOL CLoup Forest

SANCTUARY, km 2.5 old road Xalapa-Coatepec, 26 August 2011, S. Chacon, E. Utrera,

EF Tapia & G. Medrano 6293.

Gloniopsis praelonga (Schwein.) Underw. & Earle, Bull. Alabama Agricultural

Experiment Station 80: 196 (1897) FIG: 3,17

OBSERVATIONS— Gloniopsis praelonga is distinguished by its spores (18-)

20-30(-31) x 8-12(-13) um, hyaline, ovoid with obtuse ends, inequilateral,

with (5-)6-7 transverse septa and 2-3 longitudinal septa and by presenting

linear ellipsoid erumpent to superficial hysterothecia. The studied material is

consistent with the descriptions of Zogg (1962), Barr (1990a), and Boehm et al.

(2009). Previously recorded for Mexico from the State of Sonora by Méndez-

Mayboca et al. (2008), it is recorded here for the first time for the State of

Veracruz.

Hasitat— On dead branches in a forest relict of Fagus grandifolia var.

mexicana and cloud forest; 1900 m asl.

SPECIMENS EXAMINED — MEXICO. VERACRUZ: Municipality of Acatlan, ACATLAN

VOLCANO upper south, 14 November 2011, Chacon & Tapia 6618; 25 November 2011,

Chacon & Tapia 6641; Municipality of Xalapa, FRANCISCO JAVIER CLAVIJERO BOTANIC

GARDEN, Km 2.5 old road Xalapa-Coatepec, 10 March 1995, F. Tapia 1291; 28 February

1997, F. Tapia 1638; MONTEVIDEO SECTION, southeast part of the city, 30 July 2012,

F, Tapia 2595; 15 October 2012, S. Chacon 6872; LAZARO CARDENAS AVE. central green

area, ~200 m from the State Treasury Office, 22 August 2012, S. Chacon & F. Tapia 6843.

Heptameria obesa (Durieu & Mont.) Sacc., Syll. Fung. 2: 88 (1883) FIGS. 4, 5, 18

Pseudothecia subepidermal to suberumpent in the cortex, solitary or densely

confluent, subglobose to ovoid with a flat base, papilla conspicuous, completely

black, 370-600(-710) tm wide and 380-560(-600) um high; peridium

110-280 um thick, composed of thick-walled isodiametric cells, dark brown

to black. Asci (120-)130-190 x 17-22 um, claviform, bitunicate, octosporous,

the base shortly stipitate. Cellular pseudoparaphyses. Ascospores (45-)

50-72(-75) x 9-13(-14) um, fusiform, straight to slightly curved, with 6-9

septa, dictyoseptate or muriform, the average cell is dark-brown to dark-reddish

New Dothideomycetes in Mexico ... 149

and from brown pale to hyaline towards the ends with 2—3(—4) transverse septa,

slightly constricted at the septum that separates the middle portion; biseriate to

occasionally uniseriate in the ascus.

HasitTat— On dead wood (Pseudotsuga sp.) and dead branches (Baccharis

sp.), on mesquite vegetation mixed with Quercus spp. and Pseudotsuga sp. in a

Pinus-Pseudotsuga-Quercus forest; 2300-2600 m asl.

SPECIMENS EXAMINED — MEXICO. PugEBta: Municipality of Salvador el Seco, Km 62

PEROTE-PUEBLA ROAD level with height of the entrance to La Caida, 15 August 1996,

S. Chacon 5091. VERACRUZ: Municipality of Perote, PEROTE-PUEBLA ROAD 500 m after

Perote, turn 10 km to the right, 24 September 2010, S. Chacon & E. Utrera 602.

OBSERVATIONS— Heptameria obesa is characterized by the shape of its spores

(fusiform, like bull horns) with a dictyoseptate or muriform central part and

only the ends with transverse septa. The studied material coincides with the

descriptions given by Lucas & Sutton (1971), Barr (1990a), Sierra-Lopez (2006),

and Catania & Romero (2010). Heptameria has been placed in Pleosporaceae by

von Arx & Miller (1975) and in Leptosphaeriaceae by Barr (1987, 1990a). Kirk

et al. (2008) placed it within Dothideomycetes, with uncertain familial afhnity.

Heptameria obesa is known from Algeria, Argentina, Spain, USA, France,

Portugal, and Taiwan. This is the first record of the species in Mexico.

Hysterobrevium mori (Schwein.) E.W.A. Boehm & C.L. Schoch, Stud. Mycol. 64: 62

(2009) FIGS. 6, 19

= Hysterographium mori (Schwein.) Rehm

OBSERVATIONS— Hysterobrevium mori is one of the world’s most common

species and comprised the species with the second highest number of

specimens found in our study. It is distinguished from other Hysterobrevium

species primarily by its ellipsoid muriform spores of (13-)16-23(-25) x

(7—)8-10 um, with 3-5 transverse septa, as cited by Zogg (1962), Dennis (1981),

and Barr (1990a) (all as Hysterographium mori) and Boehm et al. (2009) and

Lorenzo & Messuti (2009). The species was recently reported from Sonora by

Méndez-Mayboca et al. (2010; as Hysterographium mori). Here its distribution

is extended to the states of Oaxaca, Puebla, Tamaulipas, and Veracruz.

Hasirat— On dead branches of Acacia constricta A. Gray, Helietta parvifolia

(A. Gray) Benth, and Cercidium praecox (Ruiz & Pav.) Harms, in various types

of xerophytic scrub vegetation, tropical deciduous forest, tropical forest, and

Quercus forest, from 10-1550 m asl.

SPECIMENS EXAMINED —- MEXICO. Oaxaca: Municipality of Teotitlan, VALLEY OF

TEOTITLAN, Km 73, Teotitlan-Cuicatlan road, 25 January 1991, L. Montoya 1949, 1950.

PuEBLA: Municipality of Zapotitlan Salinas, BoTANIC GARDEN OF THE BIOLOGICAL

STATION OF UNAM, 16 April 1999, S. Chacon 5129, 5131; 17 April 1999, S. Chacon

5134; 12 June 1999, S. Chacon 5151; 1.5 Km TuRN TO SANTIAGO COATEPEC, Huajuapan

Zapotitlan road, 13 December 1997, S. Chacén 5110, 5112; Flores s/n. SONORA:

150 ... Chacon, Tapia, & Esqueda

Municipality of Alamos, Huerta Vigja, 12 September 2006, S. Chacén 5736; PaLo

INJERTO, 13 September 2006, S. Chacon 5747; Mgsa DEL Trico, 14 September 2006,

S. Chacon 5769; Municipality of Fronteras, La VALDEzA, 29 August 2007, S. Chacon

5856; Municipality of Cumpas, La SELva, 30 August 2007, S. Chacon 5864. TAMAULIPAS:

Municipality of Hidalgo, Km 79 Crupap VicTorRiA-LINARES ROAD, 19 April 1996,

S. Chacén 5026, 5028. VERACRUZ: Municipality of Xalapa, PARQUE NATURA protected

natural area, 16 August 2011, S. Chacon & E. Utrera 6232; 1 November 2011, S. Chacon

& E Tapia 6584; Municipality of Actopan, La Mancua, Biological Station CICOLMA,

14 October 2004, S. Chacon 5536.

Leptospora rubella (Pers.) Rabenh., Klotzschii Herb. Viv. Mycol., Edn 2: no. 532

(1857) Fics. 7, 8, 20

= Ophiobolus rubellus (Pers.) Sacc.

Pseudothecia subepidermal, gregarious to just a few, pyriform, short

necked, sometimes broad and straight at the base resembling a flask, 450 um

high and 300 um wide, black or sometimes the host with reddish-purple dye.

Peridium 35-60 um thick, formed by sub-globose cells 5-10 um, periphery

cells dark-brown and those towards the inside, hyaline, thick wall. Asci 125-

190(-200) x (5-)6-7(-7.5) um, cylindrical, bitunicate, basal part with a short

stipe. Pseudoparaphyses abundant, filiform, hyaline, septate, 1-3 um thick,

anastomosed. Spores 120-180 x 1-1.5 um, hyaline to yellowish, cylindrical,

transversely multiseptate, irregularly spiral within the ascus.

SPECIMEN EXAMINED — MEXICO. VERACRUZ: Municipality of Xalapa. FRANCISCO

JAVIER CLAVIJERO BOTANIC GARDEN, Km 2.5 old road Xalapa-Coatepec, 2 December

2011, S. Chacon & EF. Tapia 6660.

OBSERVATIONS— The main features of this species are reddish-purple spots

that present both pseudothecia and the host and the sub-cylindrical spiral

spores with abundant septa and spore and ascus measurements that different

authors described as very variable. Furthermore, this coincides well with the

descriptions of Shoemaker (1976: as Ophiobolus rubellus), Walker (1980),

Dennis (1970), and Crous et al. (2006). Leptospora rubella is known from North

America and Europe. This is the first record of the species for Mexico.

Macrovalsaria megalospora (Mont.) Sivan., Trans. Br. Mycol. Soc. 65(3): 400 (1975)

FIGS. 9, 10, 21

= Macrovasaria leonensis (Deighton) Petr.

Ascoma solitary or sometimes confluent, unilocular to multilocular, sub-

immerse and erumpent, with traces of bark around the ascomata, subglobose-

conical, with flat base, 500-1000 um diam. and 400-750 um high, ostiolate with

a well developed papilla, which contains periphyses, black. Peridium 50-80

um thick, the outer portion covered by a crusty layer and the inner formed

by isodiametric hyaline-dark cells of intricate texture with numerous small

dark-brown to nearly black particles, Asci 250-290 x 25-30 um, cylindrical,

New Dothideomycetes in Mexico ... 151

Fics 7-14. Leptospora rubella, 7: immature and mature asci. 8: ascospores. Macrovalsaria

megalospora, 9: ascospores. 10: mature ascus. Oedohysterium insidens, 11: ascospores.

Psiloglonium clavisporum, 12: mature and immature asci. 13: ascospores. Rhytidhysteron rufulum,

14: ascospores. Scale bars: 7-9, 11-14 = 10 um; 10 = 20 um

152 ... Chacon, Tapia, & Esqueda

shortly stipitate, fissitunicate, octosporous and with a clear ocular chamber.

Pseudoparaphyses filamentous, septate, anastomosed, <2.5um thick, hyaline.

Spores 38-44 x 18-20 um, ellipsoid, finely verrucose, uniseriate, pale-brown-

dark brown, with a central septum strongly constrained and darker than the

rest of the spore, one of the ends with helmet-like structure or cover with clear

linear halos like striae covering + % of the total of the spore.

HasitTaT— On stems of Cassia fistula L. in secondary vegetation type, at 20

m asl.

SPECIMEN EXAMINED - MEXICO. VEeRAcRuz: Municipality of Cotaxtla, NEAR THE

COTAXTLA EXPERIMENTAL FIELD, 26 August 1996, A. Romero s/n.

OBSERVATIONS— Ellipsoid spores with a helmet-like cover with striae

toward one end, the presence of a single septum, and the dark-brown color

are diagnostic features of this species. Macrovalsaria leonensis was treated by

Sivanesan (1975) as asynonym of M. megalospora; consequently, Macrovalsaria

remains a monotypic genus. The Mexican material coincides well with the

description given by Sivanesan (1975), Hyde (1994), and Wang & Lin (2004).

M. megalospora is a pantropical species known from Africa, Asia, and South

America. This is the first record of the species for Mexico.

Oedohysterium insidens (Schwein.) E.W.A. Boehm & C.L. Schoch, Stud. Mycol. 64:

59 (2009) FIGS: 1b, 22

= Hysterium insidens Schwein.

OBSERVATIONS— Oedohysterium insidens is easily distinguished from other

Oedohysterium species by fusiform spores with 6-9 transverse septa and a wider

supra-median part 30-40(-45) x 10-11(-12) um. In contrast, the spore size

variability is evident. Zogg (1962) and Boehm et al. (2009) both cite spore sizes

of (20-)23-28(-38) x (5-)7-10(-13) um, while Dennis (1981) cites 21-38 x

6-12 um and Sivanesan (1984) 20-38 x 6-12 um. The species was recorded for

Sonora by Méndez-Mayboca et al. (2008; as Hysterium insidens) with spores

32-45 x 11-14 um. Here O. insidens is cited for the first time for the State of

Puebla.

Hasitat— On dead wood of Acacia constricta in xerophytic vegetation at

1500 m asl.

SPECIMENS EXAMINED - MEXICO. Pugesia: Municipality of Zapotitlan Salinas,

BIOLOGICAL STATION OF THE UNAM, 12 June 1999, S. Chacén 5142. SoNoRA:

Municipality of Fronteras, La VALDEZA, 25 August 2005, S. Chacon 5627.

Psiloglonium clavisporum (Seaver) E. Boehm, C.L. Schoch & Spatafora, Mycol. Res.

113(4): 469 (2009) Fiés.12, 13, 23

= Glonium clavisporum Seaver

Hysterothecia gregarious, superficial, arranged in parallel, linear to

navicular, some slightly curved, <3 mm long, faintly striated longitudinally,

New Dothideomycetes in Mexico ... 153

= . Se

Fics 15-24. Pseudothecia and hysterothecia. 15: Anteaglonium abbreviatum (S. Chac6n 5764):

scale bar = 1.5 mm. 16: Astrosphaeriella trochus (F. Tapia 2637): scale bar = 1.2 mm. 17: Gloniopsis

praelonga (S. Chacon 6872): scale bar = 2.0 mm. 18: Heptameria obesa (S. Chacon 5091): scale bar

= 0.5 mm. 19: Hysterobrevium mori (S. Chacon 5864): scale bar = 0.75 mm. 20: Leptospora rubella

(S. Chacon & F. Tapia 6660): scale bar = 0.4 mm. 21: Macrovalsaria megalospora (A. Romero

s/n): scale bar = 1.8 mm. 22: Oedohysterium insidens (S. Chacon 5627): scale bar = 1.2 mm.

23: Psiloglonium clavisporum (S. Chacon 5004-C): scale bar = 2.8 mm. 24: Rhytidhysteron rufulum

(S. Chacon 5859): scale bar = 0.2 mm.

with apices obtuse or acuminate, black, subiculum absent. Peridium

carbonaceous. Asci 95-120 x (10-)12-13 um, cylindrical, short stiped, hyaline

to yellowish. Pseudoparaphyses anastomosed, cylindrical 1-2 um thick, hyaline

to olivaceous-yellow. Spores (16-)18-20(-23) x (5.5-)6-6.5(—7) um, ellipsoid

154 ... Chacon, Tapia, & Esqueda

to obovoid, asymmetric, with one end rounded and one more sub-fusoid,

constrained by a central septum, hyaline, uniseriate in the ascus.

HasiTat— On decorticated wood in a semideciduous forest, 4 m asl.

SPECIMEN EXAMINED —- MEXICO. VERAcRUz: Municipality of Tampico Alto, 10

KM BEFORE THE LLANO DE BUSTOS BRIDGE, Tuxpan-Tampico road, 16 April 1996,

S. Chacon 5004-C.

OBSERVATIONS— ‘The spore size and the hysterothecial shape characterize the

species. Our specimen coincides well with the descriptions of Seaver (1925),

Lohman (1937), and Boehm et al. (2009). According to Zogg (1962), Glonium

clavisporum (= Psiloglonium clavisporum) is synonymous with G. lineare (Fr.)

De Not.; however, Boehm et al. (2009) considered them a separate species,

describing for G. lineare hysterothecia confluent and smaller spores (10-

)12-14(-18) x (4-)5-7(-8) um. Another species that could be confused with

P. clavisporum is P. chambianum (A.L. Guyot) E.W.A. Boehm & C.L. Schoch,

which differs by its wider spores (6-)8-9(-10) um; Zogg 1962; Boehm et al.

2009). Psiloglonium clavisporum is known from Africa, India, and Central

and North America. Here it is recorded for the first time for the mycobiota of

Mexico.

Rhytidhysteron rufulum (Spreng.) Speg., Anales Soc. Ci. Argent. 90: 177 (1921)

[“1920”] Fics. 14, 24

=Tryblidiella rufula (Spreng.) Sacc.

OBSERVATIONS— Rhytidhysteron rufulum is mainly characterized by discoid

apothecia, with the hymen reddish, yellowish, or even yellow-greenish and in

the hysterothecial form in young individuals and apothecial form in mature

individuals. Spores ellipsoid, 23-35 x 9-14 um, with 1-3 slightly constricted

transverse septa, which coincides with the descriptions of Dennis (1970; as

Tryblidiella rufula), Samuels & Miller (1979), Kutorga & Hawksworth (1997),

and Mendez-Mayboca et al. (2010).

Guzman (1983) cited Hysterium angustatum Alb. & Schwein. from Yucatan

and Méndez-Mayboca et al. (2008) cited H. truncatulum Cooke & Peck from

Sonora; however, our examination of both specimens on which these authors

based their descriptions indicated that they are conspecific with R. rufulum.

Salinas-Salgado et al. (2012) cited H. angustatum from Guerrero, which is the

first authentic record of this taxon for Mexico, and Méndez-Mayboca et al.

(2010) cited R. rufulum for Sonora. This paper extends its distribution to the

states of Chiapas, Guerrero, Nuevo Leon, Puebla, Queretaro, Quintana Roo,

Tabasco, Tamaulipas, Veracruz, and Yucatan.

SPECIMENS EXAMINED -— MEXICO. Cuiapas: Municipality of Ocosingo, NEAR

RUINS OF BONANPAK, 25 April 2012, EF Tapia & S. Chacon 2525; F. Tapia & S. Chacén

2531; Municipality of Tuxtla Chico, Rosario IzApA EXPERIMENTAL FIELD, Km 18

New Dothideomycetes in Mexico ... 155

Cacahuatlan-Tapachula road, 14 December 1992, S. Chacon 4633; 15 December 1995,

S. Chacén 4974. GUERRERO: Municipality of Tololoapa, Km 40 IGuALa-CruDAD

ALTAMIRANO ROAD, 2 July 1982, G. Rodriguez 234-A. NUEVO LEON: Municipality

of Santiago, KM 5 WEST OF EL CERRITO, 4 October 1980, S. Chacoén 53. PUEBLA:

Municipality of Francisco Z. Mena, MocTEezuma, along the stream on the estate of

Don Felipe Chote, 23 June 2012, S. Chacén 6790; Municipality of Zapotitlan Salinas,

UNAM BIOLOGICAL STATION BOTANICAL GARDEN, 11 August 1998, S. Chacon s/n; 18

September 1998, J. Flores s/n; 16 April 1999, S. Chacon 5128; 17 April 1999, S. Chacén

5135; 12 June 1999, S. Chacén 5141. QUERETARO: Municipality of Pinal de Amoles,

La Cugsta, Km 3 south of Escanelilla, 15 July 1983, G. Rodriguez 1973. QUINTANA

Roo: Municipality of Santa Matilde, PUERTO MORELOS TO VALLARTA ROAD, ~1 Km

from the turn to Tulum, 3 November 1981, A. Lopez 1652 (as Hysterium angustatum

in Guzman, 1983). Sonora: Municipality of Alamos, NEAR CuZALITO, 16 September

2006, S. Chacon 5812; Et Cajon, 13 September 2006, S. Chacén 5744; San Pedro, 13

September 2006, S. Chacén 5757; PRomoNTOoRIOS, 12 September 2006, S. Chacon

5711, 5730; Municipality of Cumpas, La SELva, 27 August 2005, S. Chacon 5692 (as

Hysterium truncatulum in Méndez-Mayboca et al. 2008); 30 August 2007, S. Chacon

5859. TaBasco: Municipality of Cardenas, La CHONTALPA, Ecological Reserve of the

Colegio de Posgraduados, Tabasco Campus, Km 21 Cardenas- Coatzacoalcos road,

24 April 2012, FE Tapia & S. Chacon 2509; Municipality of Cunduacan, Huimanco,

7 December 1983, R. Solano 27; Municipality of Nacajuca, EL SANDINAL, 8 February

1984, R. Solano 152; Municipality of Comalcalco, ORIENTE 2%” SECTION, 7 February

1984, R. Solano 119. TaMauLiPas: Municipality of Matamoros, ~KM 15 MATAMOROS-

VALLE HERMOSO ROAD, Via El Sendero level with the turn to the village of El Control,

17 May 2003, S. Chacén 5500, 5501; Municipality of Casas, Km 68 CruDAD VICTORIA-

SOTO DE LA MARINA ROAD, 12 June 1983, G. Rodriguez 1735; EyID0 CAMOTERO, near

Mante, 17 August 1980, A. Enock s/n. VERACRUZ: Municipality of Actopan, CICOLMA

BIOLOGICAL STATION, La Mancha, 25 August 1983, A. Sampieri 158; Municipality of

Coacoatzintla, 1 KM AFTER COACOATZINTLA, Xalapa-Naolinco road, 6 November 1984,

A. Sampieri 1104; Municipality of Dos Rios, ~5 KM BEFORE REACHING CHAVARRILLO

near the Tabiquera and the municipal landfill, 3 October 1995, S. Chacén 4941;

7 KM BEFORE CHAVARRILLO, Estanzuela-Chavarrillo road, 22 October 1999, S. Chacén

5221, 5222; Municipality of Emiliano Zapata, CHAVARRILLO AND CARRIZAL LOCAL

ROAD, 15 November 1995, S. Chacon 4962, 4959; Municipality of Hidalgotitlan, San

Car.os, Zapata and San Carlos road, 23 April 2012, S. Chacon & F. Tapia 6725, 6726;

NEAR EL MACAYAL CEMETERY, 23 April 2012, F. Tapia 2495, 6717, 6719; Municipality

of Jalcomulco, XALAPA-HUATUSCO ROAD | Km from turn to Jalcomulco, 2 July 1983,

S. Chacén 1289; 26 September 1985, S. Chacén 3096; Municipality of Minatitlan,

ROMITA 19 KM FROM MrnaTITLAN, 23 April 2012, E Tapia 2505; Municipality of

San Andrés Tlalnehuayocan, NEAR PLAN DE SEDENO, on the way to San Andres

Tlalnehuayocan, 13 October 1995, S. Chacon 4956; NEAR SAN ANTONIO HIDALGO, 14

May 2000, D. Jarvio 509; Municipality of Xico, NEAR THE WATERFALL, 22 October 1999,

S. Chacén 5209, FRANCISCO JAVIER CLAVIJERO BOTANIC GARDEN, km 2.5 old road,

Xalapa-Coatepec 11 October 1989, S. Chacon 4217; Municipality of Xalapa, PARQUE

NatuRA, Southeastern Xalapa, 30 August 2011, S. Chacdn & EF Tapia 6301; CERRO

MACUILTEPEC PaRK, Northern Xalapa, 11 October 2011, S. Chacén 6458; 25 October

2011, S. Chacon & FE. Tapia 6526. YUCATAN: Municipality of Mérida, 2 KM BEFORE

TURN TO ONCAN, Mérida-Tixkokob road, 28 October 1984, S. Chacon 2701.

156 ... Chacon, Tapia, & Esqueda

Acknowledgements

We thank the authorities of the Instituto de Ecologia, A.C. (INECOL) of Xalapa,

Veracruz, for strategic projects about Patterns of Species Diversity in Urban Zones

(grants 20035-30861; 20035-30874, and 2003 353). We also thank Biol. Elsa Ma. Utrera

Barillas for field support and logistics, technician Juan Lara Carmona for his help in

curatorial and technical activities, Manuel Escamilla for inking the drawings, and Aldo

Gutiérrez (CIAD), who prepared the final version of the pictures and text. The authors

would like to express their gratitude to Dr. Cecilia Carmaran (Buenos Aires, Argentina)

and Dr. José G. Marmolejo (Linares, N.L., Mexico) for kindly reviewing the manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/128.159

Volume 128, pp. 159-164 April-June 2014

Chaetospermum malipoense sp. nov.

from southwest China

X1AO-MING TAN »?, CHUN-LAN WANG’, JUAN CHEN’, & SHUN-XING GUO"

"Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &

Peking Union Medical College, No. 151, Malianwa North Road,

Haidian District, Beijing 100193, China;

? Guangxi Botanical Garden of Medicinal Plant, Guangxi Key Laboratory of

Medicinal Resources Conservation and Genetic Improvement, Nanning 530023, China

*CORRESPONDENCE TO: sxguo1986@163.com

ABSTRACT —A new Chaetospermum species isolated from root of Coelogyne leucantha in

Yunnan province of southwest China is described as C malipoense. It differs from other

Chaetospermum spp. by the presence of unique conidiomatal setae.

KEY worps —Ascomycota, endophytic fungi, taxonomy

Introduction

Orchidaceae is a diverse family comprising 20,000-35,000 epiphytic and

terrestrial species (Cribb et al. 2003; McCormick et al. 2004). Many studies

indicate that mycorrhizal fungi play an important role in the stages of seed

germination and growth of orchids (Zettler et al. 2004; Yam & Arditti 2009).

Additionally, non-mycorrhizal endophytic fungi associated with orchid

plants have been found to serve as potential growth promoters and source of

bioactivity substances, but studies on these fungi are very limited (Suarez et al

2006).

The genus Chaetospermum was established by Saccardo (1892). Ten species

and two varieties have been described based on morphological characteristics,

e.g., gelatinous conidiomata, conidia with tubular appendages, and

holoblastic sympodial conidiogenesis, Molecular analyses have indicated that

Chaetospermum taxa should be placed in the basidiomycete order, Sebacinales

(Rungjindamai et al. 2008). Wells & Bandoni (2001) suggested that members

of Efibulobasidium (Sebacinales) represented teleomorphs of Chaetospermum.

Kirschner & Oberwinkler (2009) also supported this phylogenetic relationship.

160 ... Tan & al.

During the investigation of endophytic fungi associated with orchid

plants in Yunnan province of southwest China, a novel species of the genus

Chaetospermum was isolated from the root of Coelogyne leucantha. We describe

it here as a new species, Chaetospermum malipoense.

Materials & methods

Malipo county is located in Wenshan zhuang and Miao minority autonomous

prefecture in the southeast of Yunnan province, China, at 22°48’-23°33’N 104°33’-105°18’E.

The plant Coelogyne leucantha grows epiphytically on the tree trunks in subtropical

evergreen forests and flowers in May-June (Flora of China Editorial Board of Chinese

Academy of Sciences 1999). In 2010, healthy roots of C. leucantha were collected in

polyethylene bags, transported to the laboratory and placed in a refrigerator at 4°C.

The plant species was identified according to Chen et al. (1999).

To isolate endophytic fungi, roots of C. leucantha washed thoroughly in running tap

water were cut into 1 cm segments and surface-sterilized in 70% ethanol for 20 s and

3% NaClO, for 5 min and rinsed in sterile distilled water 3 times. 3-5 mm segments

were separated with the help of a sterile scalpel and were placed on potato dextrose agar

(PDA) medium containing 100 g/ml oxytetracycline and 50 g/ml streptomycin (Otero

et al. 2002). Seven segments were plated per Petri dish, incubated in darkness at 25°C,

and checked daily for 4 weeks. Hyphae emerging from segments were transferred to

fresh PDA for purity and identification. The pure isolate was morphologically identified

and then confirmed by sequence analyses. After extraction of genomic DNA from

our isolate, the ITS and 18S rDNA regions were amplified with gene-specific primers

(ITS1, ITS4 — White et al. 1990; NS, SL —Inderbitzin et al. 2001) using an iCycler™M

thermal cycler (Mastercycler Gradient). The amplification cycles followed White et al.

(1990) and Inderbitzin et al. (2001), respectively. The PCR products were sequenced

by Genewiz (Beijing, China) using the same primers as for amplification. Sequences

were compared with fungal ITS and 18S rDNA sequences in GenBank using BLAST

searches. The fungal isolate was photographed and preserved in the laboratory of

Mycology, Biotechnology Center, Institute of Medicinal Plant Development (IMPLAD),

Chinese Academy of Medical Sciences, Beijing; and the culture was accessioned and

preserved in China General Microbiological Culture Collection Center (CGMCC),

Institute of Microbiology, Chinese Academy of Sciences, Beijing. Conidiation structures

of Chaetospermum malipoense from PDA cultures grown for 20 days at 25°C in darkness

were examined, measured, and photographed in a ZEISS Axio Imager Al (Germany)

microscope after mounting in 5% KOH. Measurements were made randomly from 30

cultured specimens and morphology was drawn above tracing paper by free hand and

converted into a picture using a scanner.

Taxonomy

Chaetospermum malipoense X.M. Tan & S.X. Guo, sp. nov. FIGs 1-16

MycoBank MB 801732

Differs from all other Chaetospermum spp. by its aseptate conidiomatal setae and its

larger conidia and polar appendages.

Chaetospermum malipoense sp. nov. (China) ... 161

Fics 1-12. Chaetospermum malipoense (holotype). 1. Habit. 2. Vertical section of conidioma.

3. Conidiophores and conidia. 4. Dehiscent conidiomata. 5. Monothetic conidiomatal setae.

6. Geminate conidiomatal setae. 7-10. Conidiophore branching and conidiogenous cells.

11. Guttulate on the surface of conidia. 12. Conidia with appendages. Scale bars: 2-4 = 250 um;

5-10 = 10 um; 11 = 5um; 12 = 20 um.

162 ... Tan &al.

Fics 13-16. Chaetospermum malipoense (holotype).

13, 14. Conidiomatal setae. 15. Mature conidia with appendages.

16. Conidiophore branching and conidiogenous cells.

Scale bars: 13, 14 = 50 um; 15, 16 = 10 um.

Type: China. Yunnan, Malipo County, in roots of Coelogyne leucantha W.W. Sm.

(Orchidaceae), 17 July 2010, Xiao-ming Tan (Holotype, IMPLAD 5880; ex-type culture,

CGMCC 6373; GenBank, JQ794486, JQ794487).

EryMo_Locy: malipoense, referring to the county where the specimen was sampled.

CONIDIOMATA 250-950 um diameter, opening by a regular split at the base,

firstly discrete and globose, ultimately becoming irregular spherical gregarious

and confluent, white and gelatinous when moistened. CONIDIOMATAL WALL

including many layers of non-transparent cells, pseudoparenchymatous at

the base, 200-300 um thick, textura intricata and gelatinous at the upper of

conidioma. SETAE MARGINAL, 132-362 um long, 13-23.5 um wide at the base,

light yellow, monothetic to two or three, non-septate at base. CONIDIOPHORES

arising from the lower half of the conidioma, branched and septate at base,

Chaetospermum malipoense sp. nov. (China) ... 163

smooth, firstly fusiform becoming elongate and slender towards the apex with

maturity. CONIDIOGENOUS CELLS sympodial, 2-3 in clusters. Conip1A mostly

cylindrical, smooth, hyaline, (23-)24-32(-37) x (3.5-)4.5-7.5 um (mean =

27.5 x 5 um), length/width ratio 5.6:1, abundantly guttulate. APPENDAGES

subpolar, tubular, unbranched, mostly 3-4 at each end, 14-30 um long, 1-2 um

wide at the base.

TELEOMORPH: Unknown.

COMMENTS — We place C. malipoense in Chaetospermum based on its

gelatinous conidiomata, holoblastic sympodial conidiogenesis, and cylindrical

non-septate conidia with tubular appendages (Sutton 1980; Nag Raj 1993).

Rajeshkumar et al. (2010) distinguished C. setosum from other species by

the presence of conidiomatal setae; the 1-2 septa at the base of these setae

clearly differ from the aseptate setae of C. malipoense. The nearest match for

our C. malipoense ITS sequence (JQ794486) was an Efibulobasidium albescens

sequence (AF384860; Wells et al. 2004) with a 95% similarity; however, there

are no other Chaetospermum ITS sequences in the GenBank database. The

nearest match for our C. malipoense 185 rDNA sequence (JQ794487) was a

C. camelliae sequence (EF589729; Rungjindamai et al. 2008) with a 99%

similarity. Although C. camelliae shares a similar conidial morphology with

C. malipoense, it can be distinguished by its smaller conidial size (Agnihothrudu

1962).

Acknowledgments

We are grateful to Dr. Roland Kirschner and Dr. Cheng-Lin Hou for reviewing the

manuscript and providing valuable comments. This investigation was supported by the

National Natural Science Foundation of China (30830117, 31170016, 31170314), and the

International Science and Technology Cooperation Projects of China (2011DFA31260).

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from the Western Ghats, India. Mycotaxon 113: 397-404. http://dx.doi.org/10.5248/113.397

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MY COTAXON

http://dx.doi.org/10.5248/128.165

Volume 128, pp. 165-172 April-June 2014

Chemistry and morphology of Chrysothrix candelaris in Poland,

with notes on the taxonomy of C. xanthina

SANDRA OLSZEWSKA’', ADRIAN ZWOLICKI’, & MARTIN KuKWA'*

‘Department of Plant Taxonomy and Nature Conservation &

?Department of Vertebrate Ecology and Zoology,

University of Gdansk, Wita Swosza 59, PL-80-308 Gdarisk, Poland

*CORRESPONDENCE TO: dokmak@ug.edu.pl

ABSTRACT — The chemical and morphological variation of the lichen Chrysothrix candelaris

s.l. as well as habitat requirements and distribution were studied in Poland. The species

was found to be chemically variable, but as the chemotypes do not differ in granule size,

habitat preferences, or distribution, they are treated as representing one chemically variable

taxon. ‘The size of the granules in Polish specimens is contrary to the previous reports for

C. candalaris, but the measurements are very similar to those for C. xanthina, which differs

only in distribution and habitat and is treated as a probable semi-cryptic species.

Key worps — secondary chemistry, Arthoniales

Introduction

As recently circumscribed, Chrysothrix candelaris (L.) J.R. Laundon

(Laundon 1981) is an epiphytic (rarely saxicolous) lichen occurring in Europe

and characterized by a leprose, yellow, orange yellow, or green yellow thallus

consisting of ecorticate granules and the presence of calycin (Kalb 2001,

Harris & Ladd 2008, Kukwa & Knudsen 2011). According to Kalb (2001)

the soredium-like granules (functioning as vegetative propagules) are large,

75-200 um in diam., but Harris & Ladd (2008) reported them to be much

smaller, 50-75 um in diam.

The species is very similar to C. xanthina (Vain.) Kalb (Kalb 2001), which was

resurrected from the synonymy of C. candelaris s.l., but differs in the production

of pinastric acid, granules measuring 20-50 um in diam. and occurrence in

tropical and subtropical world regions and temperate North America (Kalb

2001, Harris & Ladd 2008, Kukwa & Knudsen 2011). Although both taxa

appear easily distinguishable, some chemically deviating populations exist in

the ranges of C. candelaris and C. xanthina, and these complicate delimitation

166 ... Olszewska, Zwolicki, & Kukwa

of both species. Kalb (2001) noted some European populations containing

pinastric acid, therefore chemically similar to C. xanthina but morphologically

indistinguishable from C. candelaris. At first they were considered as probably

yet another species (Kalb 2001), but Harris & Ladd (2008) suggested that the

material could represent a chemotype of C. candelaris. Also within the range

of C. xanthina is material containing calycin and thus chemically identical

to C. candelaris but with the granule diameter characteristic for C. xanthina

(Kalb 2001). Laundon (1981) and Tonsberg (1992) also reported an additional

chemotype of C. candelaris sl. from Europe containing both calycin and

pinastric acid, but that chemotype was not studied by Kalb (2001) or Harris &

Ladd (2008).

Neither Kalb (2001) nor other authors provided studies of C. candelaris

s.l. chemotypes occurring in Europe in terms of morphological variation, and

the problem as to whether the European material represents one chemically

variable taxon or a group of species remained unresolved. Although

C. candelaris s.l. has been rather commonly reported in Poland (Faltynowicz

2003), its chemistry and morphological variation have never been studied. As

we had access to many specimens of this lichen, we studied whether Polish

chemotypes differed in granule size, distribution, and habitat preferences to

determine if Polish material of C. candelaris s.1. represented one taxon. This

is particularly important as the species is listed as critically endangered to

extinction (Cieslinski et al. 2006).

Material & methods

The material upon which the research was based consisted of 129 samples of

C. candelaris s.l., all collected in Poland and deposited in KTC, KRAM, LOD, and

UGDA. The samples were chemically analyzed with standard procedures of thin layer

chromatography (TLC) (Culberson & Kristinsson 1970, Orange et al. 2001). Secondary

metabolites were separated in solvent C and compared on each chromatogram with

control extracts of calycin, pinastric and vulpinic acids from other lichens.

In order to determine the relation between chemotypes and morphological

characters, simple granules were measured(aggregations disintegrating into smaller

granules were not considered). For this purpose, granules from 15 specimens were

measured (five of each chemotype): 51 granules from chemotype I, 57 from chemotype

I, and 37 from chemotype III (chemotype numeration follows Tonsberg 1992; see

also results). The measurements were taken in water with addition of ethanol, which

reduced the hydrophobic properties of lichen metabolites present in granules and made

the granules more easily visible; ethanol was selected, as it did not influence granule

size (checked empirically). Because the data were normally distributed and the variance

was constant, potential differences in granule sizes between chemotypes were tested by

ANOVA F-test and examined by box whisker plots in the STATISTICA v.9.1 (StatSoft

Inc. 2010), with abbreviations used in the text: N - total number of granules measured;

df — degrees of freedom; p — probability.

Chrysothrix xanthina and C. candelaris s.1. (Poland) ... 167

All examined localities are mapped according to the ATPOL system in 10 x 10 km

squares (Cieslinski & Faltynowicz 1993; see also Kukwa et al. 2002, 2010, 2012).

Results

Chemotypic variation of Chrysothrix candelaris in Poland

Three chemotypes of C. candelaris s.1. were found in Poland, the first (I) with

pinastric acid as major substance; the second (II) with calycin; and the third

(IIT) containing both calycin and pinastric acid. In chemotypes I and II vulpinic

acid was rarely also detected in minor or trace amounts. In our study we found

additional variation within chemotype II; one of the substances, calycin or

pinastric acid, was more concentrated on TLC plates than the other. That led us

to suspect that samples representing chemotype III were only mixed collections

of chemotypes I and II. Therefore, we analyzed several parts of thalli occurring

on the same piece of substrate to detect if there were chemotypes I and II on

the same bark piece, but we always got the same combination of substance

concentrations on the TLC plates from all parts of the same specimen. This

confirms that chemotype III is not a mechanical “hybrid” formed due to the

co-occurrence of individuals containing pinastric acid (chemotype I) and

calycin (chemotype IJ), but a naturally occurring chemotype producing both

chemicals.

% Chemotypes

J 5 I

[Jim

a f | li =

30 35 40 45 50

20 25 sh)

Granules diam. (in um)

PLATE 1. Chrysothrix candelaris. Percentage of granule diameter for chemotypes in Poland.

168 ... Olszewska, Zwolicki, & Kukwa

Morphological variation of Chrysothrix candelaris in Poland

The granules in 14 samples (including all three chemotypes of C. candelaris)

were mostly 30-40 um in diameter with only a few smaller and larger (PLATE 1).

Only in one sample of chemotype I were several larger granules (55 um diam.)

found. The differences in sizes of granules diameters were not statistically

significant between all chemotypes (Anova, F = 2.33, N = 144, df = 2, p =0.10).

Granules in chemotypes IJ and III were almost of the same average size (34.12

uum versus 34.44 um), whereas a slightly higher average value was found for

chemotype I (37.35 um) (PLATE 2).

Distribution and habitat requirements of Chrysothrix candelaris in Poland

The three chemotypes differ in frequencies in Poland: chemotype I is the

most frequent (94 samples), whereas II (20 samples) and III (15 samples) are

ut HI

Chemotypes

PLATE 2. Chrysothrix candelaris. Box whisker plot of differences between granule size of

chemotypes. Abbreviations: SD - standard deviation, SE - standard error.

Chrysothrix xanthina and C. candelaris s.1. (Poland) ... 169

1} EOI

LA “ot et | VE

Ht cia

Ales

dene

{et

PLATE 3. Chrysothrix candelaris. Distribution of chemotypes in Poland.

less frequent. Their distributions greatly overlap (PLATE 3), and sometimes two

different chemotypes were recorded in the same locality (but not growing side-

by-side). Only chemotype III is more restricted in distribution, being absent

from western Poland.

All chemotypes were most frequently collected on the rough bark of oaks

(mostly Quercus robur), very often in shady old-growth broad-leafed forests,

more rarely on other deciduous trees, conifers (two specimens), and wood (two

specimens), and apparently do not differ in habitat preferences.

Discussion

The chemistry of the material referred to as C. candelaris in Poland is

rather complex and our results are consistent with those of Laundon (1981)

and Tonsberg (1992) and also partly of Kalb (2001) and Harris & Ladd (2008).

170 ... Olszewska, Zwolicki, & Kukwa

However, we found an additional, previously unrecognized variation within

the third chemotype in which the ratio of the concentration of calycin and

pinastric acid varied in different samples. Actually, all specimens can be

organized to form a continuum with specimens in which only pinastric acid

was found by TLC on one side, those with only calycin detectable on the

opposite and samples with both substances in different concentrations in

between. Therefore, the boundaries between all chemotypes seem to be less

obvious than previously reported. The absence of calycin in chemotype I and

pinastric acid in chemotype II can most probably be attributed to the method

used for the analyses, and TLC may not be sensitive enough to detect traces of

those substances.

When our results of the granule measurements are compared with previous

C. candelaris treatments, one can, however, see significant discrepancies. Much

smaller granules were reported by Tonsberg (1992; (6—)12-25(-30) um diam.)

but much larger by Kalb (2001; 75-200 um diam.) and Harris & Ladd (2008;

50-75 um diam.). It is possible that the variation, at least in part, can be due

to interaction with environmental factors prevailing in different geographical

regions and habitats. To some degree, the earlier reports of large granules were

due to the misinterpretation of aggregations as simple granules, as already

pointed out by Kukwa & Knudsen (2011).

Kalb (2001) and Harris & Ladd (2008) suggested that C. candelaris might

consist of more than one species. After studying the material of C. candelaris

in Poland, we assert that it represents a morphologically uniform entity that is

chemically variable. Also the distribution patterns and habitats in Poland do

not differ between chemotypes. Under those circumstances, we do not agree

that C. candelaris is a complex of taxa and that European material containing

pinastric acid can represent separate species for which Lepra citrina Schaer. is

the oldest available name, as was discussed by Harris & Ladd (2008) and Kukwa

& Knudsen (2011).

On the other hand, the size of granules in Polish material of C. candelaris

is consistent with data for C. xanthina (Harris & Ladd 2008, Kukwa &

Knudsen 2011), which then is hardly distinguishable based on morphology

and chemistry; as pointed out above, C. xanthina contains pinastric acid, but

C. candelaris may produce that substance as well. One could regard them as

synonymous as proposed by Laundon (1981). However, as C. xanthina differs in

habitat preferences (e.g., smooth bark and fence-posts exposed to sun and rain,

often eutrophicated habitats, burned oak woodlands and chaparral, redwood

forests, bishop pine forests vs. C. candelaris usually on rough bark, rarely wood

and rocks, in humid conditions) and is found in the tropics, subtropics, and

temperate North America (vs. C. candelaris only in Europe; Kalb 2001, Harris

& Ladd 2008, Kukwa & Knudsen 2011), C. candelaris and C. xanthina could be

Chrysothrix xanthina and C. candelaris s.1. (Poland) ... 171

considered semi-cryptic species, as Vondrak et al. (2009) proposed for some

morphologically indistinguishable Caloplaca species with different distribution

ranges and ecology. We prefer this solution, as there would be a great loss

of information if the two taxa are prematurely and unjustifiably united. We

therefore propose to keep them separate until more data from all continents,

including molecular study, become available.

SELECTED SPECIMENS EXAMINED —

CHEmotyPE I. POLAND. SwigToKRzyskIE Mrs: Swietokrzyski National Park,

Chetmowa Gora, ATPOL Ee-77, on Abies alba, 1965, S. Cieslinski (KTC); KNyszyNsKA

ForestT. Budzisk nature reserve, ATPOL Cg-02, on Acer platanoides, 1999, K. Czyzewska

et al. (KTC); BreLsKa PLAIN: Bialowieska Forest, forest inspectorate Hajnéwka, forest

section no. 602D, Mysliszcze range, ATPOL Cg-64, on Quercus robur, 1982, S. Cieslinski,

Z. Tobolewski (KTC); RADOMSKA PLAIN: Kozienicka Forest, Zagozdzon nature reserve,

ATPOL De-99, on Quercus robur, 1968, S. Cieslinski (KTC); 0.75 km N of cross roads in

Zatamanek, ATPOL Df-91, on Quercus robur, 1978, A. Anusiewicz (KTC).

CHEMOTYPE II. POLAND. KRAINA WIELKICH JEZIOR MAZURSKICH: Mokre nature

reserve, by Mamry lake, ATPOL Bf-00, on Fraxinus excelsior, 1988, S. Cieslinski (KTC);

SZYDLOWSKIE FOOTHILLS: 4 km NW of Bogoria, Mostki forest district, forest section

no. 48, Ee-98, on Quercus robur, 1986, M. Chyb, K. Toborowicz (KTC); KoZrENICKA

Forest: Ponty-Deby nature reserve, ATPOL Ee-09, on Quercus robur, 2003,

S. Cieslinski (KTC); AuGUsTOWsKA PLAIN: Augustowska Forest, Perkuc, ATPOL

Bg-31, on Quercus robur, 1986, S. Cieslinski (KTC); MaAzurska P an: Kruklanka

range, by road from Marksewo to Babieta, ATPOL Be-67, on Fraxinus excelsior, 1989,

S. Cieslinski (KTC).

CuHEMOTYPE III. POLAND. Bretska PLAIN: Bialowieska Forest, Bialowieza forest

inspectorate, forest section no. 396C, ATPOL Cg-55, on Quercus robur, 1983,

S. Cieslinski, Z. Tobolewski (KTC); KoOZIENICKA FoREsT: Ponty-Deby nature reserve,

ATPOL Ee-09, on wood, 2003, S. Cieslinski (KTC); RADOMSKA PLAIN: Kozienicka

Forest, Zagozdzon nature reserve, ATPOL De-99, on Quercus robur, 1968, S. Cieslinski

(KTC).

Acknowledgments

We are grateful to Jurga Motiejunaité (Nature Research Centre, Vilnius) and James

Lendemer (The New York Botanical Garden, NY) for reviewing the manuscript and

several helpful comments and to the curators of herbaria for the loan of specimens.

Literature cited

Cieslinski S, Faltynowicz W. 1993. Note from editors. 7-8, in: S Cieslinski, W Faltynowicz (eds).

Atlas of the geographical distribution of lichen in Poland. 1. Krakow, W. Szafer Institute of

Botany, Polish Academy of Sciences.

Cieslinski S, Czyzewska K, Fabiszewski J. 2006. Red list of the lichens in Poland. 71-90, in: Z Mirek,

K Zarzycki, W Wojewoda, Z. Szelag (eds). Red list of plants and fungi in Poland. Krakow.

W. Szafer Institute of Biology, Polish Academy of Sciences.

Culberson CF, Kristinsson H. 1970. A standardized method for the identification of lichen

products. Jour. Chromatogr. 46: 85-93. http://dx.doi:10.1016/S0021-9673(00)83967-9

172 ... Olszewska, Zwolicki, & Kukwa

Faltynowicz W. 2003. The lichens, lichenicolous and allied fungi of Poland. An annotated checklist.

Biodiversity of Poland 6: 1-435. W. Szafer Institute of Botany, Polish Academy of Sciences,

Krakow.

Harris RC, Ladd D. 2008. The lichen genus Chrysothrix in the Ozark ecoregion, including a

preliminary treatment for eastern and central North America. Opuscula Philolichenum 5:

29-42.

Kalb K. 2001. New or otherwise interesting lichens. I. Biblioth. Lichenol. 78: 141-167.

Kukwa M, Knudsen K. 2011. Notes on the identity of Chrysothrix populations (Arthoniales,

Ascomycota) containing pinastric acid from southern and central California. Mycotaxon 116:

407-411. http://dx.doi:10.5248/116.407

Kukwa M, Motiejinaité J, Rutkowski P, Zalewska A. 2002. New or interesting records of

lichenicolous fungi from Poland. I. Herzogia 15: 129-139.

Kukwa M, Czarnota P, Perz P. 2010. New or interesting records of lichenicolous fungi from Poland

VIII. Herzogia 23(1): 111-119.

Kukwa M, Lubek A, Szymczyk R, Zalewska A. 2012. Seven lichen species new to Poland. Mycotaxon

120: 105-118. http://dx.doi.org/10.5248/120.105

Laundon JR. 1981. The species of Chrysothrix. Lichenologist 13(2): 101-121.

http://dx.doi:10.1017/S002428298 1000169

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

London, British Lichen Society.

StatSoft Inc. 2010. STATISTICA (data analysis software system), version 9.1. www.statsoft.com.

Tonsberg T. 1992. The sorediate and isidiate, corticolous, crustose lichens in Norway. Sommerfeltia

14; 1-331.

Vondrak J, Riha P, Arup U, Sochting U. 2009. The taxonomy of the Caloplaca citrina group

(Teloschistaceae) in the Black Sea region; with contributions to the cryptic species concept in

lichenology. Lichenologist 41(6): 571-604. http://dx.doi.org/10.1017/S0024282909008317

MY COTAXON

http://dx.doi.org/10.5248/128.173

Volume 128, pp. 173-178 April-June 2014

Two new species of Diacheopsis from China’

SHU-ZHEN YAN, MING-QUAN GUO, & SHUANG-LIN CHEN"

College of Life Sciences, Nanjing Normal University, Nanjing, 210023, China

*CORRESPONDENCE TO: chenshuanglin@njnu.edu.cn

ABSTRACT — Two new species from China in the myxomycete genus Diacheopsis are

described and illustrated. The distinctive characteristics of Diacheopsis griseobrunnea sp. nov.

are solitary and grayish brown sporocarps, sparse and uniform capillitium, and minutely

warted spores; those of Diacheopsis gigantospora sp. nov. are large spores marked with long

spines, a capillitium that forms a very loose network, and wide tubular threads. Differences

between the new species and closely related taxa are discussed. Type specimens are deposited

in the Microbial Cultures Center of Nanjing Normal University.

KEY worps —morphology, taxonomy

Introduction

The genus Diacheopsis (Stemonitaceae, Stemonitida) was established by

Meylan in 1930 with D. metallica Meyl. as the type species (Kirk et al. 2008;

Martin & Alexopoulos 1969). Sixteen species are currently known for the genus

(Hernandez-Crespo & Lado 2013), but only one species, D. mitchellii, has been

collected from China (Yamamoto et al. 2002).

Among our examinations of a number of myxomycetes obtained from

moist chamber cultures of bark samples from living trees, two specimens were

found that appeared to fit the generic concept of Diacheopsis, in that they had

sessile fruiting bodies and were limeless, non-columellate, and characterized

by an iridescent peridium. As they also differed from any known species of

Diacheopsis, they are described here as two new species.

Materials & methods

MOIST CHAMBER CULTURE: In the laboratory, the bark samples from living trees

were placed in artificial moist chambers. These chambers consisted of Petri dishes

(9 cm in diam.) lined with moistened filter paper as described by Stephenson (1989).

* Shu-Zhen Yan and Ming-Quan Guo contributed equally to this paper.

174 ... Yan, Guo, & Chen

The samples were moistened with distilled water, and excess water was poured off after

approximately 24 h. The cultures were maintained under indirect natural light at room

temperatures (23-25 °C). To maintain a moist environment, distilled water was added

to the cultures when required. The cultures were examined under the stereomicroscope

every 2 days. After 1-2 days when the myxomycete fruiting bodies were fully mature,

they were removed from the moist chambers, air-dried, and placed into a small plastic

specimen box for permanent preservation. All specimens examined are deposited in the

Microbial Cultures Center of Nanjing Normal University (MCCNNU).

MORPHOLOGICAL OBSERVATIONS: The morphology was observed using a

stereomicroscope (Jiangnan JSZ6, Nanjing, China), optical microscope (Nikon YS2-H,

China and Zeiss Axio Imager Al, G6ttingen, Germany), and scanning electron

microscope (JEOL, JSM-5610LV and JSM-5900, Tokyo, Japan). For light microscopic

observations, the preparation of slide mounts followed Martin & Alexopoulos (1969).

For SEM study, the air-dried specimens with entire and broken sporocarps were attached

to double-sided adhesive tape on sample holders of the scanning electron microscope,

and coated with platinum-palladium using a JEOL JFC-1600 Auto Fine Coater.

Taxonomy

Diacheopsis griseobrunnea Shuang L. Chen, Shu Z. Yan & M.Q. Guo, sp. nov.

MycoBank MB 805456 PLATE 1

Differs from Diacheopsis depressa by its solitary globose sporocarps, sparse capillitium

not forming a network, and smaller spores.

Type: China, Guangxi: Shangsi County, Pingguang Forest Center, alt. 250 m, on bark

of living Pinus massoniana Lamb. in moist chamber culture, collected 12 Jan. 2005,

cultured 5 May 2005, harvested 22 May 2005, S.-Z. Yan & S.-L. Chen (Holotype,

MCCNNUO0151).

ErymMo_oey: Latin griseus (gray) and brunneus (brown), referring to the grayish brown

sporocarps.

Fruiting bodies sessile and sporocarpous. Sporocarps solitary, grayish brown,

iridescent with purple and metal reflections, globose or subglobose, the base

constricted, 0.3-0.6 mm diam. Hypothallus dark brown, round, small, thin,

membranous. Peridium persistent, thin, membranous, outer surface roughened

with some irregular wrinkles. Columella absent. Capillitium sparse, connected

to the base and the peridium, a few branched, threads slender, uniform, 1-1.5

um in diam., smooth, subhyaline by transmitted light. Spore dark brown in

mass, pale violaceous brown by transmitted light, densely marked with minute

warts, globose, 6.5-8.0 um diam. Plasmodium not observed.

ComMENnts: Diacheopsis griseobrunnea is similar to D. depressa K.S. Thind

& T.N. Lakh., D. mitchellii Nann.-Bremek. & Y. Yamam., D. rigidifila S.L.

Stephenson & Nann.-Bremek., and D. synspora Nann.-Bremek. & Y. Yamam.

in the nature of the capillitium which forms a few branches but not a network.

However, D. depressa differs by its globose to subglobose to slightly elongated

Diacheopsis spp. nov. (China) ... 175

.8um

Pate 1. Diacheopsis griseobrunnea: A: Sporocarp; B. Portion of the outer peridium; C: Capillitium

and spores; D: Spore.

angular sporocarps, its abundant capillitium, and its larger spores (9-12 um

diam.; Thind & Lakhanpal 1968); D. mitchellii has larger spores (18-23 um diam.;

Nannenga-Bremekamp & Yamamoto 1983); D. rigidifila has a capillitium with

more or less fusiform thickenings in the central part (Stephenson & Nannenga-

Bremekamp 1990); and D. synspora has spores united in stable clusters of 5-7

(Nannenga-Bremekamp & Yamamoto 1986).

Diacheopsis gigantospora Shuang L. Chen, M.Q. Guo & Shu Z. Yan, sp. nov. PLATE 2

MycoBAnk MB 805455

Differs from Diacheopsis mitchellii by its very loose capillitial network, wider tubular

capillitial threads, and larger spores.

176 ... Yan, Guo, & Chen

3.0kV 8.2mm x6.00k SE{(M)

8 5mm x 5.00 3.0kV 8.4mm x20.0k SE(M}

PLATE 2. Diacheopsis gigantospora: A: Sporocarp; B. Portion of outer periduium; C: Portion of the

capillitium net; D: Branched part of the capillitium; E: Spore; F: Part of a spore.

Type: China, Yunnan: Pingbian County, Dawei Mountain National Natural Reserve,

alt. 1677 m, on bark of living Cryptomeria japonica (Thunb. ex L. f.) D. Don. in moist

chamber culture, collected 23 Nov. 2012, cultured 24 Feb. 2013, observed 29 Mar. 2013,

harvested 19 Apr. 2013, M.-Q. Guo & S.-L. Chen (Holotype, MCCNNU00706).

Erymotocy: Latin “gigant-” (gigantic) and “sporus” (spored), referring to the large

spores.

Fruiting bodies sessile and sporocarpous. Sporocarps solitary, semi-globose

to elongated, occasionally pulvinate, base wide, 0.1-0.2 mm in height, 0.3-0.5

mm in width, up to 0.7 mm in length. Sporocarps black at first, becoming dark

Diacheopsis spp. nov. (China) ... 177

brown to dull black, iridescent with blue and purple reflections when maturing.

Hypothallus dark brown, small, thin, membranous. Peridium persistent, thin,

membranous, the outer surface mostly roughened but without any distinct

ornamentation, the inner surface smooth. Columella absent. Capillitium

sparsely branched and with few interconnections, forming a very loose

network, threads white or pallid but limeless, straight, tubular, 2-5 um in diam.

and sometimes expanded at the junctions and marked with a few bead-like

thickenings on the larger branch. Spores black in mass, dark brown to black

brown by transmitted light, obviously marked with evenly-distributed spines

up to over 1 um long, globose, (17.5-)20.6-26.2(-28) um diam. including

spines. Plasmodium black.

ComMENTs: Diacheopsis species having large spores include D. insessa (G. Lister)

Ing (14-19 um; Martin & Alexopoulos 1969), D. kowalskii Mar. Mey. & Poulain

(15-18.5 um; Meyer & Poulain 1998), and D. mitchellii (18-23 um; Nannenga-

Bremekamp & Yamamoto 1983). However, the spore sizes in these three species

are smaller than those in D. gigantospora. In addition, D. insessa differs by its

densely clustered sporocarps (sometimes with weak stalks) and a purplish

capillitium; D. kowalskii has gregarious and pulvinate sporocarps, a bicolored

capillitium, and flattened threads bearing darker axillary expansions; and

D. mitchellii has sporocarps in small groups, capillitium of simple or sparsely

dichotomous threads ca. 0.5 um diam. and not forming a network, and spores

densely covered with evenly-distributed nail-headed warts or blunt spines.

Several other species of Diacheopsis have wide capillitium threads,

including D. kowalskii, D. nannengae G. Moreno et al., D. pauxilla Mar. Mey.

& Poulain, D. reticulospora Mar. Mey. & Poulain, D. serpula Kowalski, and

D. synspora. From D. gigantospora, D. nannengae differs by its larger sporocarps,

brown capillitium, and smaller spores (Moreno et al. 1989); D. pauxilla by its

gregarious and greater sporocarps, and smaller spores (Meyer & Poulain 1998);

D. reticulospora by its plasmodiocarpous sporocarps, brown capillitium, and

spores densely and irregularly reticulated (Meyer & Poulain 1990); D. serpula

by its plasmodiocarpous sporocarps, purple-brown threads, and smaller

spores (Meyer & Poulain 1998); and D. synspora by its gregarious and larger

sporocarps, dichotomously branched capillitial threads and spores united in

stable clusters of 5-7 (Nannenga-Bremekamp & Yamamoto 1986).

Acknowledgments

We are sincerely grateful to Prof. S. L. Stephenson of the University of Arkansas in

the United States, and Prof. Wen-Ying Zhuang of the Institute of Microbiology, Chinese

Academy of Sciences, China for their critical review and suggestions on improving the

manuscript. We also deeply thank Mr. Jun- Yong Dai and Tian-Peng Song for their helps

in collecting bark samples, and Mr. Xing-Guang Xie for his cartographic assistance. This

178 ... Yan, Guo, & Chen

study was supported by National Nature Science Foundation of China (no. 31170014)

and the fund of Key Program of Natural Science of Jiangsu Higher Education Institutions

of China (12KJA180004).

Literature cited

Hernandez-Crespo JC, Lado C. 2013. An online nomenclatural information system of Eumycetozoa.

Real Jardin Botanico de Madrid, Spain. www.nomen.eumycetozoa.com (2014).

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th ed. CAB International, Wallingford. 771 p.

Martin GW, Alexopoulos CJ. 1969. The Myxomycetes. University of Iowa Press, Iowa. 561 p.

Meyer M, Poulain M. 1990. Une nouvelle espéce nivale du genre Diacheopsis. Beitr. Kenn. Pilze

Mitteleurop. 6: 35-38.

Meyer M, Poulain M. 1998. Diacheopsis kowalskii et Diacheopsis pauxilla - deux novelles especies

de myxomycetes. Bull. Fed. Myc. Dauphine-Savoie 150: 27-37.

Moreno G, Illana C, Heykoop M. 1989. Contribution to the study of the myxomycetes in Spain I.

Mycotaxon 34: 623-635.

Nannenga-Bremekamp NE, Yamamoto Y. 1983. Additions to the myxomycetes of Japan I. Proc.

K. Ned. Akad. Wet. Ser. C, 86: 207-242.

Nannenga-Bremekamp NE, Yamamoto Y. 1986. Additions to the myxomycetes of Japan II. Proc.

K. Ned. Akad. Wet. Ser. C, 89: 217-240.

Stephenson SL. 1989. Distribution and ecology of myxomycetes in temperate forests. 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, Nannenga-Bremekamp NE. 1990. Five new species of myxomycetes from North

America. Proc. K. Ned. Akad. Wet. Ser. C, 93: 187-196.

Thind KS, Lakhanpal TN. 1969 [“1968”]. The myxomycetes of India XXII. Mycologia 60:

1080-1085. http://dx.doi.org/10.2307/3757292

Yamamoto Y, Chen SL, Degawa Y, Hagiwara H. 2002. Myxomycetes from Yunnan Province, China.

Bull. Nat. Sci. Mus., Tokyo, Ser. B, 28: 61-76

MY COTAXON

http://dx.doi.org/10.5248/128.179

Volume 128, pp. 179-183 April-June 2014

Australohydnum dregeanum new to Italy

ALESSANDRO SAITTA , MARIA LETIZIA GARGANO,

RICCARDO COMPAGNO, & GIUSEPPE VENTURELLA

Department of Agricultural and Forest Sciences, University of Palermo,

Viale delle Scienze 11, I-90128 Palermo (Italy)

* CORRESPONDENCE TO: alessandro.saitta@unipa.it

ABSTRACT — We report on the first finding in Italy (and the third in Europe) of

Australohydnum dregeanum. A detailed description of the macro- and micromorphological

features, the ecological data, and some taxonomic remarks concerning this taxon are

provided. The bluish tint in the centre of the hymenophoral surface and the non-cystidiform

aspect of the marginal hyphae are previously undescribed characters.

KEY worps — Corticiaceae, Phanerochaetaceae, wood-inhabiting fungi, urban biodiversity

Introduction

Wood decay fungi are widely distributed in forest ecosystems of Italy. Their

importanceis highlighted by the fundamental role they play in the decomposition

of woody debris (Saitta et al. 2011) and by the high level of diversity reported

from broad-leaved and conifer woods (Venturella et al. 2011). Although

in recent years the mycological exploration in forest ecosystems of Italy has

definitely increased, it is still possible to find infrequent and/or rare lignicolous

species growing as saprotrophs or parasites on fallen trunks, branches, stumps,

etc. (Saitta & Melo 2012). Such is the case with Australohydnum dregeanum,

recently collected and identified as new to Italy (a third record from Europe).

Three species are included in Australohydnum Julich (Phanerochaetaceae):

A. castaneum, A. dregeanum, and A. griseofuscescens; we follow here the

synonymy outlined by Melo & Hjortstam (2002), which treats A. dregeanum

and A. griseofuscescens as conspecific.

Materials & methods

The basidiomata were collected inside the “Parco della Favorita’, belonging to the

Natural Reserve of “Monte Pellegrino’, a wide green area within the city of Palermo

180 ... Saitta & al.

(Sicily). The macroscopic identification was carried out on fresh basidiomata while the

microscopic features were observed under a Leica microscope DMLB on dried specimens

using a 0.3% KOH solution and cotton blue in lactic acid. The spore measurements were

based on 50 observations carried out on fresh and dried basidiomata. The nomenclature

follows MycoBank (http://www.mycobank.org), and the description of A. dregeanum

is based on the authors’ observation of the collected basidiomata. The specimens

(SAF 001, SAF 002) are kept in the fungal dried reference collection of the mycological

herbarium (under establishment) of the new Department of Agricultural and Forest

Sciences (activated by the University of Palermo on 2014).

Taxonomy

Australohydnum dregeanum (Berk.) Hjortstam & Ryvarden,

Syn. Fung. 4: 61. 1990. PLT

= Corticium dregeanum Berk., London J. Bot. 5: 3. 1846.

= Lopharia dregeana (Berk.) P.H.B. Talbot, Bothalia 6: 57. 1951.

= Hydnum griseofuscescens Reichardt, Verh. Zool.-Bot. Ges. Wien 16: 374. 1866.

= Austrohydnum griseofuscecens (Reichardt) Jiilich, Persoonia 10: 138. 1978.

= Irpex vellereus Berk. & Broome, J. Linn. Soc., Bot. 14: 61. 1873.

= Oxyporus vellereus (Berk. & Broome) A. Roy & A.B.

De, J. Mycopathol. Res. 36: 41. 1998.

= Irpex purpureus Yasuda ex Lloyd, Mycological Notes 50: 715. 1917.

Basidiomata annual, resupinate to effuse-reflexed, forming patches up to

5 cm wide, and laterally confluent and effused <15 cm, cracked when dried.

The hymenial surface is bluish in the center when fresh, gradually lilac to the

margin; brownish when dried, with some areas soft lilac; more or less warted,

warts <2.5 mm long, differently anastomosed. Margin evident, sterile, distinctly

white, slightly tomentose <4 mm broad. The basidiomata can be easily separated

from the substrate only when dried. Flesh thin, white.

Hyphal system pseudodimitic. Hyphae simple septate. Hymenial hyphae

prevalently thin-walled. Subicular hyphae thick-walled, 4-5.5 um wide.

Marginal hyphae 4-5 um wide, hyaline, more or less thick-walled, frequently

branched, sometimes encrusted at the apex. Basidia hyaline, (22-—)25-35(-40) um

long, sinuous-clavate, simple septate at the base, with sterigmata 3.5-4.5 um

long. Spores (4.5-)4.8-5.3(-5.8) 2.5-2.8(-3.2) wm, ellipsoid, hyaline,

smooth, thin-walled, inamyloid, cyanophilous. Skeletocystidia very abundant,

cylindrical <125 um long and 4.5-6.5(-7) wide, apices obtuse, more or less

encrusted, originating from pseudoskeletal hyphae and rarely projecting

beyond the hymenium <15 um, incrusted part 25-45 um.

SPECIMENS EXAMINED: ITALY, Sicity, PALERMO, Parco della Favorita, 38.15457°N

13.34628°E, 44 m a.s.l., Mediterranean maquis with scattered plants of Eucalyptus

sideroxylon A. Cunn. ex Woolls, on fallen trunk of E. sideroxylon, 4 Apr 2012, coll.

A. Saitta (SAF 001); 5 Oct 2012, coll. A. Saitta (SAF 002).

Australohydnum dregeanum new to Italy... 181

PLATE 1. Australohydnum dregeanum (SAF 002). a) Fresh basidioma; b) Dried basidioma;

c) Skeletocystidium; d) Basidia; e) Spores.

Discussion

Australohydnum dregeanum was collected on fallen trunk of E. sideroxylon

in the clearings of a Mediteranean maquis. The vegetation is also characterized

182 ... Saitta & al.

by scattered plants and shrubs of Quercus ilex L., Ulmus minor Mill., and

Pistacia lentiscus L. mixed with conifers, Cedrus atlantica (Endl.) Manetti ex

Carriere, C. deodara (Roxb. ex D. Don) G. Don, and Cupressus sempervirens L.

Eucalyptus sideroxylon was introduced in reforestations approximately 60

years ago, when the local Department of Forestry began an intense planting of

exotic species on the Sicilian territory. Eucalyptus wood is a new substrate, and

the Mediterranean maquis is a new habitat for A. dregeanum in Europe. Melo &

Hjortstam (2002) reported A. dreageanum on fallen branches of Olea europaea

L. var. europaea in a holm oak forest (Quercus rotundifolia Lam.) with a small

abandoned olive grove and lusitanic oaks (Q. faginea Lam.).

Owing to the presence of abundant encrusted skeletocystidia and simple

septate generative hyphae, A. dregeanum is rather easily identified. The

morphological features of the A. dregeanum collected in Sicily are similar to

those reported by Melo & Hjortstam (2002) for specimens collected in Portugal.

The main difference in the features of the Sicilian specimens are the bluish tint in

the centre of hymenophoral surface (not reported in any previous description)

and the non-cystidiform aspect of the marginal hyphae. The bluish tint in the

centre of hymenophoral surface is clearly visible in the fresh specimens.

The genus Australohydnum in Europe includes only one taxon, A. dregeanum,

which was collected in Portugal by Melo & Hjortstam (2002) as new to

Europe. The species is also known from Australia (Reichardt 1866, as Hydnum

griseofuscescens; Reid 1956, as Irpex vellereus), New Zealand (Buchanan &

Ryvarden 2000), Japan (Lloyd 1917, as Irpex purpureus), South Korea (Lim et

al. 2005), Sri Lanka (Berkeley & Broome 1873, as Corticium dregeanum), India

(De 1998, as Oxyporus vellereus; Tiwari et al. 2010], South Africa (Berkeley

1846, as Corticium dregeanum; Talbot 1951, as Lopharia dregeana), Israel (Tura

et al. 2010), and Russia (Zmitrovich et al. 2006).

Lim & Jungh (2003) described a new species, Irpex hydnoides Y.W. Lim

& H.S. Jung, microscopically quite similar to A. dregeanum but which has a

different hymenophoral configuration. They analyses did not place Irpex

vellereus within the I. hydnoides-I. lacteus clade but grouped it instead (with

100% support) with a sequence of A. dregeanum, supporting the synonymy of

A. dregeanum and I. vellereus. Their A. dregeanum-I. vellereus clade was sister

to the group that included members of the genus Phanerochaete.

The presence of A. dregeanum in Italy is noteworthy, considering its

fragmented European distribution. This new finding also supports Sicily as a

“hot spot” of biodiversity for aphyllophoroid fungi (Saitta et al. 2011).

Acknowledgments

The authors wish to thank Dr. Ireneia Melo (Portugal) and Dr. Georgios I. Zervakis

(Greece) for kindly revising the manuscript. Thanks are also due to Dr. Shaun Pennycook

Australohydnum dregeanum new to Italy ... 183

(New Zealand) for nomenclatural review and Dr. Cristiano Losi for critical comments

provided.

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MY COTAXON

http://dx.doi.org/10.5248/128.185

Volume 128, pp. 185-194 April-June 2014

Anamorphic fungi of the Atlantic Forest of southern Bahia:

new records and Dactylaria pseudomanifesta sp. nov.$

DiLz—E MARIA ARGOLO MAGALHAES’ , EDNA DoRA MARTINS NEWMAN Lwz’,

ALBERTI FERREIRA MAGALHAES', MARCOS VINICIUS OLIVEIRA DOS SANTOS’,

FLAVIA RODRIGUES BARBOSA}, LARISSA ARGOLO MAGALHAES‘,

& JosE Luiz BEZERRA***

‘Centro de Pesquisas do Cacau, Comissdo Executiva do Plano da Lavoura Cacaueira - CEPLAC,

Rod. Ilhéus-Itabuna, km 22, Ilhéus, BA, 45662-000, Brazil

*Depto. de Micologia, Universidade Federal de Pernambuco - UFPE,

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

*Instituto de Ciéncias Naturais, Humanas e Sociais, Universidade Federal de Mato Grosso,

Avenida Alexandre Ferronato, 1200, Setor Industrial, Sinop, MT, 78557-267, Brazil

‘Universidade Estadual de Santa Cruz - UESC,

Rod. Ilhéus-Itabuna, km 16, Ilhéus, BA, 45662-000, Brazil

* CORRESPONDENCE TO: dilze.argolo@yahoo.com.br

ABSTRACT — A new species, Dactylaria pseudomanifesta, and nine other species newly

recorded from Brazil (Beltraniella fertilis, Chaetosphaeria innumera, Dictyosporium oblongum,

Idriella acerosa, Inesiosporium longispirale, Pseudobeltrania macrospora, Scolecobasidium

tropicum, Sporidesmium coffeicola, and Triscelophorus curviramifer) are presented. The

specimens were recovered from the litterfall of Parinari alvimii, Manilkara maxima, and

Harleyodendron unifoliolatum, native trees of the Atlantic Forest in Bahia State, Brazil.

Key worps — conidial fungi, taxonomy, Chrysobalanaceae, Sapotaceae, Fabaceae

Introduction

The Atlantic Forest is the third largest biome in Brazil (after the Amazon

and Cerrado) and the biome of richest biodiversity on the planet (Conservac¢ao

Internacional do Brasil 2003). It is estimated that this biome contains about

20,000 plant species (of which 8,000 are endemic), accounting for 2.7% of the

total number of plants of the world (Myers et al. 2000). Taxonomic studies

on decomposer fungi in the Atlantic Forest are necessary, especially because

‘Part of the dissertation submitted by the first author for a Master’s Degree in Genetics and

Molecular Biology, Universidade Estadual de Santa Cruz - Ilhéus BA, Brazil

186 ... Magalhaes & al.

it is a biome with a significant biologic diversity which presents a extremely

high level of endemism, more strong in the south of Bahia State (Conservacao

Internacional do Brasil et al. 2000).

New studies on anamorphic fungi in the Atlantic Forest in Brazil have

expanded the knowledge of the geographical distribution of species and

diversity of fungi present in this biome (Castafeda-Ruiz et al. 2001, 2003;

Calduch et al. 2002; Gusmao et al. 2005, 2008; Cruz et al. 2007; Marques et

al. 2008; Barbosa et al. 2008, 2009; Magalhaes et al. 2011) but not as much in

southern Bahia.

Here we present records of conidial fungi isolated from the fallen litter

of the Atlantic Forest of south Bahia, including a new species, Dactylaria

pseudomanifesta, and nine other taxa not previously reported from Brazil.

Materials & methods

The study was done in three conservation units of the biome Atlantic Forest of

southern Bahia, Brazil: (1) Ecoparque, in the municipality of Una; (2) Parque Estadual

Serra do Conduru (PESC), in the municipalities of Urucuca, Itacaré and Ilhéus; and (3)

Reserva Capitao, in the municipality of Itacaré.

Three specimens of three representative endemic plant species well distributed in

the Atlantic Forest of southern Bahia were identified in the field: Parinari alvimii Prance

(Oiti cumbuca; Chrysobalanaceae); Manilkara maxima T. D. Penn. (Massaranduba;

Sapotaceae), and Harleyodendron unifoliolatum Cowan (Fabaceae). Surveys were

conducted on four occasions during the period from December 2007 to July 2008.

Plants were identified, georeferenced, and marked.

A 50 x 50 cm square (0.25 m’) was thrown over the litter of these trees and one

leaf was randomly collected at a time, totaling ten leaves/plant under different stages

of decomposition. In the Laboratory of Fungal Diversity of Centro de Pesquisa do

Cacau (CEPEC) — Comissao Executiva do Plano da Lavoura Cacaueira (CEPLAC),

samples were washed for one hour in running water and incubated in moist chambers

(Castafieda-Ruiz et al. 2006).

After 72 hours of incubation the material was observed under the stereomicroscope

and the light microscope and periodically reviewed for 30 days. All microfungi

specimens were studied in a semi-permanent mounting medium (PVL resin + polyvinyl

alcohol lactophenol; Trappe & Schenck 1982) and identified. Colonized leaves and semi-

permanent mounts were converted into exsiccates and deposited in CEPEC herbarium.

Taxonomy

Dactylaria pseudomanifesta J.L. Bezerra & D.M.A. Magalh., sp. nov. PLATE 1

MycoBAnk 807666

Differs from Dactylaria manifesta by its larger brown conidia.

Type: Brazil. Bahia: municipality of Una, Ecoparque, 15°10’01.7”S 39°03'14.2”W, on

decomposing leaves of Manilkara maxima, 10/5/2008, D.M.A. Magalhaes s.n. (CEPEC

1459).

Dactylaria pseudomanifesta sp. nov. (Brazil) ... 187

YL /

PiatE 1. Dactylaria pseudomanifesta: A, B, F Conidiophores (and conidia); C-E. Conidia.

Scale bars: A, B, C, E, F= 15 um; D = 3 um.

EryMoLocy: Latin pseudomanifesta, referring to the similarity with Dactylaria

manifesta.

Co.oniss effuse, grayish, thin, without superficial mycelium and restricted

immersed mycelium. CONIDIOPHORES simple, cylindrical, brown, septate, with

188 ... Magalhaes & al.

smooth walls, 55-188 x 5-6 um. CONIDIOGENOUS CELLS terminal, integrated,

denticulate 35-40 x 5-6 um; DENTICLES apical, in number of three to five, 1-2

um long. Conip1a obclavate to turbinate, 3-septate, light brown with a hyaline

distal cell, 17-25 x 3.5-4 um.

ADDITIONAL SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF URUCUCA,

Parque Estadual Serra do Conduru, 14°23’07.7”S 39°04’43.6”W, on decomposing leaves

of Harleyodendron unifoliolatum, 12/7/2008, D.M.A. Magalhaes s.n. (CEPEC 1417).

ComMENTs — ‘This species resembles Dactylaria manifesta R.F. Castaneda

& W.B. Kendr., which differs by its smaller (14-20 x 3-3.5 um) and totally

colourless conidia (Castafieda-Ruiz & Kendrick 1991).

Beltraniella fertilis Heredia, R.M. Arias, M. Reyes & R.F. Castafieda, Fungal

Diversity 11: 100 (2002). PLATE 2A,B

Cotontgs hypophyllous, effuse. IMMERSED MYCELIUM with setae upright,

straight, single or clustered, thick walled, smooth to slightly warty, acute, dark

brown, 90-240 x 3-6 um, bulbous base. CONIDIOPHORES macronematous,

setiform, single or in small groups, straight, septate, simple or branched at the

apex, 97-337 x 40-8 um. SEPARATING CELLS unicellular, hyaline, ellipsoid,

fusiform, 6-8 x 2.5-5 um. CONIDIOGENOUS CELLS terminal polyblastic,

integrated, pale brown. Conrp1A arising from conidiogenous cells or separating

cells, rostrate, truncated at the base, smooth, olivaceous, with a hyaline

transverse band above the equatorial zone, 15-23 x 5-8 um.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF UNA, Ecoparque,

15°10’44.6”S 39°02’06.4’W, on decomposing leaves of Parinari alvimii, 7/12/2008,

D.M.A. Magalhaes s.n. (CEPEC 1410).

DISTRIBUTION — Described from Mexico in dead leaves of Mangifera indica (Heredia et

al. 2002). First report for Brazil.

ComMENTS — The specimen analyzed agrees with the description presented by

Heredia et al. (2002).

Chaetosphaeria innumera Berk. & Broome ex Tul. & C. Tul., Select. Fung. Carpol.

2: 252 (1863). PLATE 2C

Cotontss effuse, black. PERITHECIA unobserved. CONIDIOPHORES single or

fasciculate, septate, dark brown, smooth, up to 200 um long, 4-6 um wide, often

percurrent. Conrp1A clustered, ellipsoid to oblong, unicellular, olivaceous,

2.5-5 x 2-3 um.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF URucuca, Parque

Estadual Serra do Conduru, 14°23’08.4”S 39°04’37.9”W, on decomposing leaves of

Harleyodendron unifoliolatum, 7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1463).

DISTRIBUTION — Europe including Great Britain (Ellis 1976). First report for Brazil.

ComMENTS — The specimen analyzed agrees with the description presented by

Ellis (1976). The teleomorphic phase was not observed.

Dactylaria pseudomanifesta sp. nov. (Brazil) ... 189

PLATE 2. Fungi found in the Atlantic Forest of south Bahia — Beltraniella fertilis:

A. Conidiophore branched at the apex; B. Conidia and conidiophores. Chaetosphaeria

innumera: C. Fasciculate conidiophores and clustered conidia. Dictyosporium oblongum:

D, E. Oblong conidia. Idriella acerosa: F. Terminal conidiogenous cells; G. Semilunar conidia.

Scale bars: A, B= 15 um; C= 5 um; D, F, G = 10 um; E = 30 um.

Dictyosporium oblongum (Fuckel) S. Hughes, Can. J. Bot. 36: 762 (1958). PLATE 2D,E

CoLonigs effuse. Conrp1 oblong or irregular, 32.5-37.5 x 10-12.5 um,

olivaceous, usually with 3-4 rows of cells of the same or different size, generally

strongly constricted at the septa; cells 5 um wide at 10-11 per row.

190 ... Magalhaes & al.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF URucucA, Serra do

Condurt (PESC), 14°23’08.4’S 39°04’37.9” W, on decomposing leaves of Harleyodendron

unifoliolatum, 7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1419).

DISTRIBUTION — Recorded in Europe and North America (Ellis 1971; GBIF 2014). First

report for Brazil.

ComMENTs — ‘The specimen analyzed agrees with the description presented

by Ellis (1971).

Idriella acerosa R.F. Castafieda & W.B. Kendr., Univ. Waterloo Biology Series 35:

62 (1991). PLATE 25,G

MyceELIuM superficial composed of brown, branched, septate smooth,

hyphae 15-20 um wide. CONIDIOPHORES mononematic, unbranched, erect,

straight, 4-6-septate, brown with almost colorless apex, 55-123 x 4-5 um.

CONIDIOGENOUS CELLS terminal sympodial with diminutive denticles almost

colorless at the apex, 15-22 x 2.5-5 um. Conip1A falciform with acute ends,

non-septate, dry, smooth and colorless, 10-15 x 1.5 um.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF URucucA, Serra do

Condurt (PESC), 14°23’08.4’S 39°04’37.9” W, on decomposing leaves of Harleyodendron

unifoliolatum, 7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1421).

DISTRIBUTION — Described from Cuba, on dead leaves of Samanea saman and

Nectandra coriacea (Castafieda-Ruiz & Kendrick 1991). Mexico (GBIF 2014). First

report for Brazil.

COMMENTS — ‘The specimen analyzed agrees with the description presented

by Castafieda-Ruiz & Kendrick (1991).

Inesiosporium longispirale (R.F. Castafieda) R.F. Castafieda & W. Gams, Nova

Hedwigia 64: 486 (1997). PLATE 3A

Myce ium superficial with septate, branched, smooth, brown to dark brown,

hyphae 1-5 x 3-5 um wide. ConrpiopHoREs undifferentiated, intercalated

with CONIDIOGENOUS CELLS, short, subulate, dark brown, monoblastic, 12-20

x 5-5.5 um. CONIDIA acrogenous, single, coiled, twisted, usually attenuated and

truncated at the base, multi-septate, olivaceous to pale brown, with 3-10 spirals,

smooth walled, 70-196 x 3-7 um, spirals 50-65 um x 18-36 um, filaments

3-7 um wide, olivaceous to pale brown, basal cells dark brown to brown.

SPECIMENS EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF UNA, Ecoparque,

15°10’01.7”S 39°03’14.2”W, on decomposing leaves of Manilkara maxima, 05/10/2008,

D.M.A. Magalhaes s.n. (CEPEC 1461); MUNICIPALITY OF ITACARE, Reserva Capitao,

14°2116.7”S 39°03’23.3”W, on decomposing leaves of Manilkara maxima, 7/12/2008,

D.M.A. Magalhaes s.n. (CEPEC 1556); 05/10/2008, D.M.A. Magalhaes s.n. (CEPEC

1405); MUNICIPALITY OF UruGUCA, Serra do Conduru, 14°23’07.7”S 39°04’43.6” W, on

decomposing leaves of Harleyodendron unifoliolatum, 05/10/2008, D.M.A. Magalhaes

s.n. (CEPEC 1462); 14°23’08.3”S 39°04’47.0”W, on decomposing leaves of Manilkara

maxima, 7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1553); 14°23’07.7”S 39°04’43.6’W,

Dactylaria pseudomanifesta sp. nov. (Brazil) ... 191

PLATE 3. Fungi found in the Atlantic Forest of south Bahia — Inesiosporium longispirale: A. Coiled

conidia. Pseudobeltrania macrospora: B, C. Conidia and conidiophores. Scolecobasidium

tropicum: D. Conidia and conidiophore; E. Septate conidia. Sporidesmium coffeicola: F. Straight or

curved conidiophores and conidia; G. Obpyriform septate conidia. Triscelophorus curviramifer:

H. Obclaviform conidia. Scale bars: A, F = 10 um; B, D, H = 15 um; C = 30 um; E, G=5 um.

05/10/2008, D.M.A. Magalhaes s.n. (CEPEC 1503); 12/21/2007, D.M.A. Magalhaes s.n.

(CEPEC 1425); 14°23’06.6”S 39°04’44.7”W, on decomposing leaves of Parinari alvimii,

7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1555).

192 ... Magalhaes & al.

DISTRIBUTION — Found on dead leaves of Calophyllum antillanum and Clusia rosea, in

Cuba (Castafieda-Ruiz & Gams 1997). First report for Brazil.

CoMmMENTS — The specimens analyzed are similar to the description presented

by Castafieda-Ruiz & Gams (1997).

Pseudobeltrania macrospora Heredia, R.M. Arias, M. Reyes & R.F. Castafieda,

Fungal Diversity 11: 103 (2002). PLATE 3B,C

Cotonizes epiphyllous, effuse. MyceLrum immersed. CONIDIOPHORES

macronematous, single or fasciculate, simple, septate, erect, brown, darker at the

apex, 45-88 x 5-8 um, with basal cell radiated-lobed, up to 20 um in diameter.

CONIDIOGENOUS CELLS polyblastic, terminal, cylindrical, pale brown, 15-30 x

6-7 um. Conrp1a holoblastic, single, rhomboid, with acute base and apiculate

apex, olivaceous, smooth, 30-45 x 15-20 um, with hyaline equatorial band.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF ITACARE, Reserva Capitao,

14°21'17.6”S 39°03’23.4’W, on decomposing leaves of Parinari alvimii, 7/12/2008,

D.M.A. Magalhaes s.n. (CEPEC 1412).

DISTRIBUTION — Mexico (Heredia et al. 2002). First report for Brazil.

ComMENTsS — The specimen fits the description presented by Heredia et al.

(2002).

Scolecobasidium tropicum Matsush., Matsushima Mycological Memoirs 3: 15

(1983). PLATE 3D,E

Cotonies effuse. HYPHAE SUPERFICIAL hyaline olivaceous, sparse.

CONIDIOPHORES mononematous, macronematous, scattered, simple, cylindrical,

0-2-septate, brown, with denticulate ends elongating sympodially, 10-15 x 5 um.

ConipiA fusiform, 2-septate, non-constricted at the septa, 17-20 x 3-4 um,

smooth, sub-hyaline.

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF UNA, at Ecoparque,

15°10'01.8”S 39°03’13.5”W, on decomposing leaves of Manilkara maxima, 05/10/2008,

D.M.A. Magalhaes s.n. (CEPEC 1418).

DISTRIBUTION — Federated States of Micronesia, Canada, and Thailand, on dry petioles

of Cocos nucifera. Republic of China (GBIF 2014). First report for Brazil.

ComMENTS — The specimen analyzed agrees with the description presented by

Matsushima (1983).

Sporidesmium coffeicola M.B. Ellis, More Dematiaceous Hyphomycetes: 87 (1976).

PLATE 35,G

MyceELium immersed. CONIDIOPHORES straight or curved, septate, brown,

smooth, 75-110 x 3-5 um, percurrent. Conip1A obpyriform, 2-3-septate, 15-

25 x 7-9 um, 2-2.5 um at the base, with the two lower cells dark brown and

upper cells sub-hyaline.

Dactylaria pseudomanifesta sp. nov. (Brazil) ... 193

SPECIMEN EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF URuGuCA, at Serra do

Condurt, 14°23’07.0’S 39°04’43.9’W, on decomposing leaves of Harleyodendron

unifoliolatum, 7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1409).

DISTRIBUTION — Found in Iran (coffee leaves), Japan, and Mexico (GBIF 2014). First

report for Brazil.

ComMENTs — ‘The specimen analyzed agrees with the description presented

by Ellis (1976).

Triscelophorus curviramifer Matsush., Matsushima Mycological Memoirs 7: 70

(1993). PLATE 3H

ConipiA obclavate, 1-3-septate, 19-37 x 1.5-3.0 um, with two obclavate

branches, curved at the base, 0-1-septate, 9-22 x 1.4-2.5 um.

SPECIMENS EXAMINED — BRAZIL. BAHIA: MUNICIPALITY OF UNA, Ecoparque,

15°10’44.6”S 39°03’06.4’W, on decomposing leaves of Parinari alvimii, 7/12/2008,

D.M.A. Magalhaes s.n. (CEPEC 1503); MUNICIPALITY OF UruGuca, Serra do Condurt

(PESC), 14°23’08.3”S 39°04’47.0’W, on decomposing leaves of Manilkara maxima,

7/12/2008, D.M.A. Magalhaes s.n. (CEPEC 1502).

DISTRIBUTION — Peru on rotten leaves (Matsushima 1993), and Venezuela (Smits &

Cressa 2005).

ComMENTS — The specimens analyzed agree with the description presented by

Delgado-Rodriguez et al. (2004).

Acknowledgements

The authors would like to thank Comissao Executiva do Plano da Lavoura

Cacaueira (CEPLAC) and the Conselho Nacional de Desenvolvimento Cientifico e

Tecnoldgico (CNPq) for financial support and to Katia Maria T. Bezerra (CEPLAC)

for the teachings and collaboration in the laboratory practices. Also sincere thanks to

Drs. Alvaro Figueredo dos Santos and Nadja S. Vitoria for the pre-submission review of

this manuscript.

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MY COTAXON

http://dx.doi.org/10.5248/128.195

Volume 128, pp. 195-202 April-June 2014

A new slender species of Aureoboletus

from southern China

MING ZHANG??, Tal-Hut1 LI»?’, & BIN SONG?

'School of Bioscience & Bioengineering, South China University of Technology,

Guangzhou, 510006, China

?Guangdong Institute of Microbiology, Guangzhou 510070, China &

State Key Laboratory of Applied Microbiology,

South China (The Ministry—Province Joint Development)

“CORRESPONDENCE TO: mycolab@263.net

ABSTRACT — Aureoboletus tenuis sp. nov. is described from Guangxi Province, China. It is

characterized by slender basidiomes with a glutinous, wrinkled, and light brown or reddish-

brown to dark brown pileus. LSU sequence analysis confirms placement of the new species

in Aureoboletus. A colour plate showing fruitbodies in situ and illustrations of microscopic

elements are provided.

Key worpD — Basidiomycota, Boletales, taxonomy

Introduction

Aureoboletus was circumscribed by Pouzar (1957) (with A. gentilis (Quél.)

Pouzar as type species) based on the following morphological features: small

to medium sized basidiomes, an often viscid (but sometimes subtomentose)

pileus, golden yellow or bright yellow (even when dried) tubes and pores,

and a subequal or fusoid viscid to glutinous stipe that usually tapers to the

base and is not distinctly reticulate or glandulose. Singer (1942), who had

established Xerocomus sect. Auripori for X. gentilis [= Aureoboletus gentilis]

and other species with golden yellow or bright olive-gold pores, a viscid pileus,

and a gelatinous layer on the stipe (with or without a weak veil), later (1947)

recombined it as Pulveroboletus sect. Auripori, which was to form the basis of

Pouzar’s new genus, Aureoboletus. Although not recognized as an independent

genus by some mycologists (Corner 1972, Singer 1986, Sutara 2005),

Aureoboletus has been accepted by others (Watling 1970, Pilat & Dermek 1974,

Watling & Largent 1976, Alessio 1985, Dermek 1987, Li & Song 2002, Yang et

196 ... Zhang, Li, & Song

al. 2003, Kirk et al. 2008). Recent molecular studies on boletes now support

Aureoboletus as an independent genus (Binder 1999, Binder & Hibbett 2006,

Dentinger et al. 2010, Feng et al. 2012).

Of the twelve species and varieties placed within Aureoboletus (Klofac 2010),

only two (A. reticuloceps M. Zang et al., Aureoboletus thibetanus (Pat.) Hongo

& Nagas.) have been reported from China (Patouillard 1895, Zang et al. 1993,

Ying & Zang 1994, Yang et al. 2003). However, one, A. reticuloceps, has been

transferred to Boletus based on morphological and molecular evidence (Wang

& Yao 2005, Dentinger et al. 2010, Feng et al. 2012). Recently, a new species,

described here as Aureoboletus tenuis, was discovered in Guangxi Province,

southern China.

Materials & methods

Specimens were photographed and annotated in the field and then dried in an electric

drier. Type specimens were deposited in the Fungal Herbarium (GDGM) of Guangdong

Institute of Microbiology, Guangzhou, China. Macroscopic description is based on

fresh and dried specimens, field notes, and colour photographs. Colour notations follow

Kornerup & Wanscher (1978). For descriptions of microscopic characters (including

pileipellis and stipitipellis), tissue sections were revived and examined in 5% potassium

hydroxide (KOH) or 1% Congo Red. Thirty basidiospores and 10 basidia were randomly

selected from a mature specimen and measured in KOH; Q = spore length/width ratio;

Q_, = mean ratio.

DNA was extracted from dried specimens using the Sangon Fungus Genomic

DNA Extraction kit (Sangon Biotech Co., Ltd., Shanghai, China) according to the

manufacturer's instructions. The large subunit (nLSU) region was amplified by PCR,

using primers LROR and LR5 (Pinruan et al. 2010). The amplified products were

determined by electrophoresis on 1% agarose gel against a known standard DNA

marker and directly sequenced in Beijing Genomic Institute (BGI). The LSU sequence

was submitted to GenBank. These and reference sequences from GenBank were used

in phylogenetic analysis after being edited and aligned using Clustal 1.81 (Thompson et

al. 1997) and MEGAS5.1 (Tamura et al. 2011). The dataset was analyzed with maximum

parsimony by PAUP* 4.0b10 (Swofford 2003) following He & Li (2013).

Taxonomy

Aureoboletus tenuis T.H. Li & Ming Zhang, sp. nov. FIGS 1, 2

MycoBank MB 804773

Differs from Aureoboletus auriporus var. novoguineensis by its smaller fruitbodies and

shorter basidiospores and from A. thibetanus by its wrinkled to shallowly reticulate

pileus surface and the absence of appendiculate veil remnants.

Type: China, Guangxi Province, Maoershan National Nature Reserve, 25°50’N 110°19’E,

1387 m alt., on soil in a broadleaved forest dominated by Fagaceae (Cyclobalanopsis sp.),

16 July 2012, Ming Zhang (Holotype, GDGM 42601, GenBank KF534789).

Erymo_oey: the specific epithet indicates the slender habit of the new species.

Aureoboletus tenuis sp. nov. (China) ... 197

Fic 1. Aureoboletus tenuis (Holotype GDGM 42601). Basidiomes. Scale bar = 20 mm.

BaSIDIOMES slender. P1LEus 20-35 mm broad, hemispherical when young,

becoming broadly convex to nearly plane in age, strongly glutinous when fresh,

distinctly wrinkled to irregularly and shallowly reticulate, brown to reddish-

brown (6D8-8D8, 8E8) at center and gradually paler outwards, deep orange

(5A8-6A8), orange (5A7-6A7), orange-yellow (4A8-4A7), light yellow to

pale yellow (3A4-4A4) at margin, with a slightly incurved edge when young.

CONTEXT 3-4 mm thick at the centre of the pileus, thinner at pileus margin,

soft, white to yellowish-white (1A1-1A2), more or less brown beneath the

pileipellis, slightly changing pinkish-white (7A2-9A2) to pale red (7A3-9A3)

when exposed. TuBEs slightly depressed around stipe, light yellow to greenish-

yellow (2A5, 2B5), 8-10 mm deep, unchanging on bruising. Pores 0.8-1

mm in diam., roundish to angular, somewhat compound and relatively larger

around the stipe, concolorous with tubes. StrpE 40-70 x 3-7 mm, central,

cylindrical, sometimes hollow and usually tapering towards the base, greyish-

red to brownish-orange (7C4-7D4), smooth, without reticulation, sometimes

longitudinally striate, gelatinous or strongly viscid, especially when young and

wet, with a white (1A1) basal mycelium. Opor none. TasTE mild.

BasIDIOsPoREs (10—)11-12 x 4-5 um, Q = (2.4—)2.5-2.8(-3), Q. = 2.75 +

0.29, n = 30, ellipsoid, smooth, yellowish to yellowish-brown in 5% KOH, thin-

walled. Basrp1a 4-spored, 20-26 x 8-10 um, clavate, yellowish-white to hyaline

in 5% KOH, yellow to yellowish-brown in Melzer’s. STERIGMATA 2.5-3.5 um

198 ... Zhang, Li, & Song

Aldi A! i

Hin \ ty |

NISNUAY USS

TINA RS Zeeeee—

WO Nt Wt i reSS

—

Fic. 2. Microscopic features of Aureoboletus tenuis (Holotype GDGM 42601).

A. Basidia and pleurocystidia; B. Basidiospores; C. Pileipellis; D. Stipitipellis.

Scale bars: A, B = 10 um; C, D = 20 um.

long. PLEUROcysTIDIA 40-53 x 11-15 um, fusiform, thin-walled, yellowish-

white to hyaline in 5% KOH. CueiLocystipia 42-52 x 11-15 um, similar to

pleurocystidia in shape and size. HYMENOPHORAL TRAMA Subparallel to nearly

bilateral, yellowish-white to hyaline in 5% KOH, composed of branching

hyphae 6-9 um wide, hardly to only slightly gelatinized. PILEIPELLIs in young

material usually an ixotrichodermium consisting of loosely and vertically

arranged, frequently septate, thin-walled hyphae 6-12 um in diam., tending to

collapse in mature and dried specimens, yellowish white to hyaline in 5% KOH.

STIPITIPELLIS ixotrichodermial, producing branching hyphae 7-13 um wide,

with slightly swollen tips. CAULOcysTIDIA not observed. CLAMP CONNECTIONS

absent in all tissues.

ECOLOGY & DISTRIBUTION — Solitary or scattered on soil in a broad-leaved

forest dominated by Fagaceae (Cyclobalanopsis sp.). Known only from the type

locality.

ComMENtTs — ‘The diagnostic features of the new species include the small

and slender basidiomes, glutinous and wrinkled pileus, cylindrical viscid

Aureoboletus tenuis sp. nov. (China) ... 199

Suillus pictus AY684154

Zangia roseola JQ928623

42601 Aureoboletus tenius

Aureoboletus thibetanus KJ907380

Aureoboletus thibetanus KF 112420

Aureoboletus thibetanus AY700189

Aureoboletus auriporus JQ003659

Aureoboletus auriporus HQ161871

Aureoboletus gentilis KF 112344

Aureoboletus gentilis DQ534635

Aureoboletus citriniporus KF030298

Aureoboletus moravicus KF 112421

Aureoboletus roxanae KF030311

Aureoboletus innixus KF030240

Aureoboletus innixus KF030239

Retiboletus aff nigerrimus JQ928627

Boletus ornatipes AF456805

Austroboletus dictyotus JX901138

Leccinum aurantiacum AF 139689

Rossbeevera griseovelutina HQ693882

Phylloporus bellus AY612817

Xerocomus subtomentosus AF 139716

Chalciporus piperatus DQ534648

Boletus edulis AF456816

Xanthoconium purpureum HQ161864

Bothia castanella DQ867115

Heimiella retispora AFO050650

Boletellus ananas AY612799

Strobilomyces floccopus AY684155

Porphyrellus porphyrosporus DQ534643

Borofutus dhakanus JQ928617

98 Spongiforma squarepantsii HQ724509

Tylopilus felleus EU522827

Xerocomus chrysenteron AF514808

Fic. 3. The phylogenetic tree obtained from Maximum Parsimony analysis of LSU sequences of

species of Boletales. Suillus pictus is selected as outgroup. Parsimony bootstrap values >50% are

shown.

200 ... Zhang, Li, & Song

and sometimes hollow stipe, and relatively short basidiospores. This unique

combination of characters easily separates A. tenuis from other Aureoboletus

species.

Aureoboletus auriporus var. novoguineensis (Hongo) Klofac and A. thibetanus

also have a glutinous and wrinkled pileus. However, A. auriporus var.

novoguineensis differs by its larger and more robust fruitbodies, pale reddish-

brown stipe, and longer basidiospores (11.5-15.5 um; Hongo 1973, Klofac

2010), and A. thibetanus differs by its chestnut-brown, rusty-brown to pale

brown, more distinctly reticulate-alveolate pileus that is ornamented by strongly

gelatinized veil remnants hanging at margin, longer basidiospores (9.0-15.0 x

4.0-5.5 um), and thin-walled cystidia with a refractive substance on the surface

(Patouillard 1895, Yang et al. 2003, Klofac 2010).

Aureoboletus tenuis is easily differentiated from similarly coloured taxa such

as A. flaviporus (Earle) Klofac and A. roxanae (Frost) Klofac, as A. flaviporus

exhibits more robust fruitbodies and larger basidiospores (11-18 x 4-6 um;

Earle 1904, Both 1993, Bessette et al. 2000, Klofac 2010) while A. roxanae has a

dry and broader pileus (<90 mm) and sturdier stipe (<70 x 16 mm; Frost 1874,

Smith & Thiers 1971, Bessette et al. 2000, Klofac 2010).

The phylogenetic tree (Fic. 3) clusters all sampled Aureoboletus species

in a clade with 73% bootstrap support and A. tenuis and A. thibetanus in a

subclade with a 80% bootstrap value. This well-supported lineage indicates

that the new taxon A. tenuis is sister to A. thibetanus, but the blast result with

nrLSU sequence shows that the two species share only 97% max identity. Thus,

A. tenuis is phylogenetically and morphologically distinct from all the sampled

species.

Acknowledgments

Sincere thanks are expressed to Dr. Beatriz Ortiz-Santana (US-Forest Service,

Northern Research Station, Center for Forest Mycology Research, USA) and Dr. Matteo

Gelardi (Bracciano, Italy) who reviewed the manuscript and provided invaluable

suggestions. Thanks are also given to Dr. Vladimir Antonin (Moravian Museum,

Department of Botany, Brno, Czech Republic) for supplying and translating literature,

to Miss Chao-qun Wang and Dr. Wang-qiu Deng for their valuable suggestions on

the manuscript. Acknowledgement is sincerely expressed to Mr. Cheng-shu Qiu for

his assistance. This study was supported by grants from the National Natural Science

Foundation of China (No. 31170026, 31101592, 31070024, 31093440).

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Binder M, Hibbett DS. 2006. Molecular systematics and biological diversification of Boletales.

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Both EE. 1993. The boletes of North America: a compendium Buffalo Museum of Science, Buffalo,

New York.

Corner EJH. 1972. Boletus in Malaysia. Singapore Publishing House.

Dentinger BTM, Ammirati JF, Both EE, Desjardin DE, Halling RE, Henkel TW. Moreau P,

Nagasawa E, Soytong K, Taylor AF, Watling R, Moncalvo J, Mclaughlin DJ. 2010. Molecular

phylogenetics of porcini mushrooms (Boletus section Boletus). Molecular Phylogenetics and

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Dermek A. 1987. Fungorum rariorum icones coloratae. Pars XVI. Boletes III. Cramer, Berlin.

Earle FS. 1904. Mycological studies II. Bulletin of the New York Botanical Garden 3: 289-312

Feng B, Xu J, Wu G, Zeng NK, Li YC, Bau T, Kost GW, Yang ZL. 2012. DNA sequence analyses

reveal abundant diversity, endemism and evidence for Asian origin of the porcini mushrooms.

PLoS ONE 7(5): 37567. http://dx.doi:10.1371/jo urnal.pone.0037567

Frost CC. 1874. Catalogue of boleti of New England, with descriptions of new species. Bulletin of

the Buffalo Society of Natural Sciences 2: 100-105.

He SH, Li HJ. 2013. Pseudochaete latesetosa and P. subrigidula spp. nov. (Hymenochaetales,

Basidiomycota) from China based on morphological and molecular characters. Mycological

Progress 12: 331-339. http://dx.doi.org/10.1007/s11557-012-0838-6

Hongo T. 1973. Enumeration of the Hygrophoraceae, Boletaceae and Strobilomycetaceae. Bulletin of

the National Science Museum Tokyo 16(3): 537-557.

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CABI Publishing, UK.

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treatment. Osterreichische Zeitschrift fur Pilzkunde 19: 133-174.

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of selected agaric families. Nova Hedwigia 28: 569-636.

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thibetanus. Mycotaxon 86: 283-290.

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Acta Mycologica Sinica 12(4): 275-282.

MYCOTAXON

http://dx.doi.org/10.5248/128.203

Volume 128, pp. 203-204 April-June 2014

Nomenclatural revision of four Peziza species

GIANFRANCO MEDARDI', ANGELA LANTIERI’, & GABRIELE CACIALLI?

'Via Giuseppe Mazzini 21, I-25086 Rezzato (Brescia), Italy

?Via Novaluce 38, I-95030 Tremestieri Etneo (Catania), Italy

°Via Goito 25, I-57127 Livorno, Italy

*CORRESPONDENCE TO: *gianfranco.medardi@virgilio.it

ABSTRACT — Three replacement names and a new combination are published for four Peziza

species whose current names in Peziza are illegitimate later homonyms: Peziza delilei nom.

nov. (= P. viridifusca Delile ex De Seynes), Peziza elaeocarpa comb. nov. (= P. olivacea Quél.),

Peziza pauli nom. nov. (= PB martinii Donadini), and Peziza queletii nom. nov. (= P. ampelina

Quél.).

Key worps — Pezizaceae, Pezizales, nomina nova

Some species of European Peziza Dill. ex Fr. : Fr. have no legitimate names

in Peziza and are known currently by illegitimate names (later homonyms;

McNeill et al. 2012, Article 53.1). This work resolves these nomenclatural

irregularities.

Peziza delilei Medardi, Lantieri & Cacialli, nom. nov.

MycoBank MB808495

= Peziza viridifusca Delile ex De Seynes, in De Seynes, Veg. Inf. 3: 83. 1886,

nom. illegit., non Peziza viridifusca Fuckel 1870.

= Humaria viridifusca Sacc., Syll. Fung. 8: 149. 1889.

= Galactinia viridifusca (Sacc.) Boud., Hist. Class. Discom. Eur.: 48. 1907.

Erymo toy: after Alire Raffenau Delile, who discovered the species.

Peziza elaeocarpa (Sacc.) Medardi, Lantieri & Cacialli, comb. nov.

MycoBank MB808494

= Peziza olivacea Quél., Bull. Soc. Bot. Fr. 25(4): 291. 1879 [“1878”], nom. illegit.,

non Peziza olivacea Batsch 1783.

= Humaria elaeocarpa Sacc., Syll. Fung. 8: 148. 1889.

204 ... Medardi, Lantieri, & Cacialli

Peziza pauli Medardi, Lantieri & Cacialli, nom. nov.

MycoBANK MB808493

= Peziza martinii Donadini, Bull. Soc. Linn. Provence 35: 171. 1985 [“1984”],

as “martini, nom. illegit., non Peziza martinii Pers. : Fr. 1822.

Erymotoey: from Paulus, the Latinized baptismal name of Paul Martin, to whom the

species was originally dedicated.

Peziza queletii Medardi, Lantieri & Cacialli, nom. nov.

MycoBank MB808492

= Peziza ampelina Quél., Grevillea 8(47): 116. 1880, nom. illegit.,

non Peziza ampelina Pass. 1874.

= Plicaria ampelina Rehm, Rabenh. Krypt.-FL., Ed. 2, 1(3): 1003. 1894.

= Galactinia ampelina (Rehm) Boud., Hist. Class. Discom. Eur.: 47. 1907.

ErymMo toy: after Lucien Quélet, who discovered the species.

ComMENTs: We do not accept the synonymy of P. ampelina Quel. with P arenaria

Osbeck 1762 (as reported by Species Fungorum, http://www.speciesfungorum.

org/GSD/GSDspecies.asp? RecordID=148680); the few characters of Osbeck’s

original diagnosis of P arenaria delineate a brown, cup-shaped, sabulicolous

species with a split margin, not a typical violaceous, saucer-shaped fungus,

which is P. ampelina of Queélet.

Acknowledgments

The authors thank Prof. G. Consiglio and Dr. C. Losi for critically reviewing the

manuscript and Dr. Shaun Pennycook for his precious advice and observations.

Literature cited

McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp

S, Marhold K, Prado J, Pru@homme van Reine WF; Smith GE, Wiersema JH, Turland NJ. 2012.

International Code of Nomenclature for algae, fungi, and plants (Melbourne Code), adopted

by the Eighteenth International Botanical Congress, Melbourne, Australia, July 2011. Regnum

Vegetabile 154. 208 p. http://www.iapt-taxon.org/nomen/main.php

MYCOTAXON

http://dx.doi.org/10.5248/128.205

Volume 128, pp. 205 April-June 2014

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT — Mycotaxon is pleased to announce the posting of a new species distribution

list by Calaga, Silva, & Xavier-Santos, who present the coprophilous and other dung-related

fungi recorded in Brazil. This brings to 112 the number of free access mycobiotas now

available on the Mycotaxon website: http://www.mycotaxon.com/resources/weblists.html

SOUTH AMERICA

Brazil

FRANCISCO JUNIOR SIMOES CALAGA, NATHAN CARVALHO DA SILVA, & SOLANGE

XAVIER-SANTOS. A checklist of coprophilous fungi and other fungi recorded

on dung from Brazil. 22 p.

ABSTRACT — A review of the literature published between 1919 (the earliest known

record) and 2013 has made it possible to confirm the occurrence of 209 species of

coprophilous fungi (sensu lato) in Brazil, which are distributed in 259 records in 12

states of the Federation, with Pernambuco being the State most represented. The

phylum most found was Ascomycota (117 species), followed by Zygomycota (54),

Basidiomycota (25), Myxomycota (11), Oomycota (1) and Proteobacteria (1).

MY COTAXON

http://dx.doi.org/10.5248/128.207

Volume 128, pp. 207-208 April-June 2014

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 128

Anacraspedodidymum C.R. Silva, R.E. Castafieda & Gusmao, p. 12

Anacraspedodidymum aquaticum C.R. Silva, R.E Castaieda & Gusmao, p. 12

Anacraspedodidymum hyalosporum (Bhat & W.B. Kendr.) R.F. Castaneda,

C.R. Silva & Gusmao, p. 14

Arachnophora combuensis J.S. Monteiro, R.E. Castafieda & Gusmao, p. 128

Atelocauda sakuraguiae Salazar-Yepes & A.A. Carvalho, p. 18

Aureoboletus tenuis T.H. Li & Ming Zhang, p. 196

Chaetospermum malipoense X.M. Tan & S.X. Guo, p. 160

Dactylaria pseudomanifesta J.L. Bezerra & D.M.A. Magalh., p. 186

Diacheopsis gigantospora Shuang L. Chen, M.Q. Guo & Shu Z. Yan, p. 175

Diacheopsis griseobrunnea Shuang L. Chen, Shu Z. Yan & M.Q. Guo, p. 174

Diplococcium variegatum S.S. Silva, Gusmao & R.E. Castaneda, p. vi

[validation of Diplococcium variegatum S.S. Silva, Gusmao & R.F. Castaneda,

Mycotaxon 127: 60 (2014), nom. inval., ICN (2012) Art. 42.1]

Hymenochaete biformisetosa Jiao Yang & S.H. He, p. 139

Marasmius midnapurensis A.K. Dutta, P. Pradhan & K. Acharya, p. 119

Muscodor strobelii Meshram, S. Saxena & N. Kapoor, p. 96

Peziza delilei Medardi, Lantieri & Cacialli, p. 203

= Peziza viridifusca Delile ex De Seynes 1886, nom. illegit.

Peziza elaeocarpa (Sacc.) Medardi, Lantieri & Cacialli, p. 203

= Peziza olivacea Quel. 1879 (‘1878’), nom. illegit.

Peziza pauli Medardi, Lantieri & Cacialli, p. 204

= Peziza martinii Donadini 1985, nom. illegit.

Peziza queletii Medardi, Lantieri & Cacialli, p. 204

= Peziza ampelina Quél. 1880, nom. illegit.

Puccinia caricis-araliae Kakish. & Q. Wang (neotypified, epitypified), p. 75

= Aecidium araliae Sawada ex S. Ito & Muray. (1943)

non Puccinia araliae Ellis & Everh. (1891)

Ravenelia costae Salazar-Yepes & A.A. Carvalho, p. 20