IV ... MYCOTAXON 131(3)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-ONE (3) — TABLE OF CONTENTS

COVER SECTION

REVIEWETS <6 walt coe wee e afrcet nv eae tied ees see bot ly elie ala eens Sar 4 vii

SULDIISSTOMTOCCUTOS A alay yh nig yeniatags ge Petree te neg ae Oo ae ee viii

ROUGE ETO AOE» hos oe) Ba bn hb Sih, eae OR eae Abie tte’ ix

RESEARCH ARTICLES

Periconia notabilis sp., nov. and a new record and notes

on the genus in Thailand CHARUWAN CHUASEEHARONNACHAI,

SAYANH SOMRITHIPOL & NATTAWUT BOONYUEN 491

New host records of rust fungi (Pucciniales) from Pakistan

B. ALI, Y. SOHAIL & A.S. Mumtaz 503

New records of myxomycetes from México

Marcos LizARRAGA, GABRIEL MORENO & MARTIN EsQUEDA 511

Marthamyces coronadoae sp. nov. in a

Fagus grandifolia var. mexicana forest from Hidalgo State, México

TANIA RAYMUNDO, RICARDO VALENZUELA & MARTIN ESQUEDA 521

Lithothelium bermudense sp. nov., a new saxicolous lichen

from Bermuda FRANZ BERGER, SCOTT LAGRECA & ANDRE APTROOT 527

Harmoniella campanaensis sp. nov. from central Chile

HuGo Maprip, DANIELA TORRES & ViCTOR SILVA 535

Rosellinia hainanensis sp. nov. and three Rosellinia species

new to China We! Li & Lin Guo 541

Phaeomonilia nanningensis sp. nov. and a

new Craspedodidymum record from southern China Cuun-Line YANG,

Jin- YE WANG, JI- WEN X1A, YING-RuI Ma,

JIAN-MEI GAO & XIU-GUO ZHANG 547

Pseudoacrodictys from southern China: P ambigua sp. nov.

and a new record JtAN-MEI Gao, JIN-YE WANG, CHUN-LING YANG,

JI-WEN X1A, YING-Rur Ma & X1IU-GUO ZHANG 553

Capsicispora mycophila gen. & sp. nov. from southern China

Jin- YE WANG, CHUN-LING YANG, JI-WEN XIA,

YING-Ru1 Ma, JIAN-MEI Gao, YU-MEI CAI & XIU-GUO ZHANG 559

Diploschistes tianshanensis sp. nov., a corticolous species

from Northwestern China GULIBAHAER ABABAIKELI, ABDULLA ABBAS,

SHOU-YU Guo, ANIWAER TUMIER & REYIMU MAMuTI 565

JULY-SEPTEMBER 2016... V

Sporidesmiella lushanensis and S. jiulianshanensis spp. nov.

and a new record from China Jian Ma 575

Corynesporopsis obclavata and Stanjehughesia jiangxiensis spp. nov.

from Lushan Mountain, China JIAN Ma 583

New records of Rhizocarpon from Hengduan Mountains, China

WEI-CHENG WANG, ZHAO-JIE REN, LU-LU ZHANG & ZUN-TIAN ZHAO 589

Ellisembia henanensis sp. nov. and two new hyphomycete records

from central China JI- WEN X1A, JIN-YE WANG,

CuuN-LING YANG, ZHUANG LI & X1U-GUO ZHANG 597

Thozetella coronata and T. ypsiloidea spp. nov.

from the Brazilian Amazon forest JOSIANE SANTANA MONTEIRO,

RAFAEL F. CASTANEDA-RuIz & Lufs FERNANDO PASCHOLATI GusMAO 605

Long-hidden in Thaxter’s treasure trove: Laboulbenia camerunensis sp. nov.

parasitic on African Curculionidae

TRISTAN W. WANG, DANNY HAELEWATERS & DONALD H. PFISTER 613

Erysiphe acantholimonis sp. nov. on Acantholimon hedinii

J1a-GE SONG, B1ao Xu, SHAN-HE ZHANG,

ZHEN-YU ZHAO, SUNG-EUN CHO & HYEON-DONG SHIN 621

Tubulicrinis martinicensis sp. nov., a corticioid species

from Martinique (French West Indies) GERALD GRUHN,

Nits HALLENBERG & REGIS COURTECUISSE 631

Roccella elisabethae with lichenicolous Arthonia follmanniana

in Turkey MEHMET GOKHAN HALIcI & ARIFE MERVE KAHRAMAN 639

Diaporthe henanensis sp. nov., an endophytic fungus in

Ziziphus jujuba from China Yr YANG, Yu-X1A Guo,

Ya-KuNn ZHANG, HaAI-YAN Wu & MENG ZHANG 645

Notes on rust fungi in China 1. Autoecious life cycle of

Puccinia tatarinovii on Prenanthes JING-XIN JI, QI WANG,

ZHUANG LI, Yu Li & MAKOTO KAKISHIMA 653

Gymnopus ramulicola sp. nov., a pinkish species

from southern China SHuU-FanG Dena, Tat-Hut Lt,

ZI-DE JIANG & BIN SONG 663

Hemithecium hainanense sp. nov. with

a checklist and key to Hemithecium species from China

ZE-FENG JtA, MEI-FANG LI & XIN ZHAO 671

First record of Faurelina in the Neotropics | ROGER FAGNER RIBEIRO MELO,

ANDREW N. MILLER & LEONOR Costa Malta 679

Anaverticicladus uncinatus gen. & sp. nov. from decaying leaves

from Brazil PHELIPE M.O. Costa, MARCELA A. BARBOSA,

ELAINE MALOSSO & RAFAEL F, CASTANEDA-RUIZ 687

VI ... MYCOTAXON 131(3)

Morphology and phylogeny of Cladosporium subuliforme, causing yellow leaf

spot of pepperin Cuba — Beatriz RAMos-Garcfa, TOMAS SHAGARODSKY,

MARCELO SANDOVAL-DENIS, YARELIS ORTIZ, ELAINE MALOSSO,

PHELIPE M.O. CosTA, J. GUARRO, DAVID W. MINTER, DAYNET SOSA,

SIMON PEREZ-MARTINEZ & RAFAEL F. CASTANEDA-RuiIz 693

Boletus durhamensis sp. nov. from North Carolina

BEATRIZ ORTIZ-SANTANA, ALAN E. BESSETTE & OWEN L. MCCONNELL 703

Diorygma fuscum sp. nov. from China JIAN Li & ZE-FENG Jia 717

BOOK REVIEWS AND NOTICES LORELEI L. NORVELL 723

Mushrooms of the Redwood Coast—a comprehensive guide to the

fungi of coastal northern California Noau SIEGEL & CHRISTIAN SCHWARZ

Fungi of Northern Europe—Vol. 5. The genus Mycena s.1.

ARNE ARONSEN & THOMAS LASSQE

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN VOLUME 131(3) 733

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-ONE (2)

MYCOTAXON for APRIL-JUNE 2016, (I-xII + 263-490)

was issued on August 12, 2016

JULY-SEPTEMBER 2016...

REVIEWERS — VOLUME ONE HUNDRED THIRTY-ONE (3)

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 quarter.

Vladimir Antonin

Alan W. Archer

José Luiz Bezerra

Meredith Blackwell

Uwe Braun

Jose Francisco Cano-Lira

R.F. Castaneda-Ruiz

Cvetomir M. Denchev

Jorge Rat Deschamps

Adam Flakus

Genevieve Gates

Josepa Gené

Roy E Halling

Liu-Fu Han

David L Hawksworth

Peter Johnston

Santosh Joshi

Alfredo Justo

Bryce Kendrick

Kerry Knudsen

De-Wei Li

Hua-Jie Liu

José G. Marmolejo

Eric H.C. McKenzie

Armin MeSi¢

Josiane S. Monteiro

Akira Nakagiri

Lorelei L. Norvell

Shaun R. Pennycook

Sergio Pérez-Ortega

Liliane Petrini

Walter P. Pfliegler

Huzefa A. Raja

Roger Rosentreter

Amy Rossman

Isabel Salcedo

Roger G. Shivas

Steven L. Stephenson

Susumu Takamatsu

Merje Toome- Heller

Larissa Vasilyeva

Martin Westberg

Xiu-Guo Zhang

VII

vill ... MYCOTAXON 131(3)

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

subscription articles are offered.

JULY-SEPTEMBER 2016... IX

RICHARD P. KORE, MYCOLOGICAL GIANT [1925-2016]—Eight Saturdays ago,

on August 20, this journal lost its mentor and co-founder, who passed away

after a bout of pneumonia at the age of 91. Although a death occurring in the

tenth decade of life should not be unexpected, the news came as a shock to the

editorial office and left us feeling decidedly adrift in far too vast mycological

sea. MycoTaxon had hoped to publish a memoriam to Dick in this issue, but

as two months proved too short a time to assemble the materials to do him

justice, we hope to close out 2016 by publishing in our October-December

issue reminiscences of our spirited scientist, nomenclaturalist, international

forayer, actor, and self-proclaimed curmudgeon.

We invite any and all of you to send in your photographs, anecdotes, and

musings to give Dick the rousing send-off he so richly deserves. Please send

your reflections—before December 15, if possible—to MycoTaxon in care of

my office via editor@mycotaxon.com.

MYCOTAXON 130-3 presents 30 papers by 114 authors (representing 19

countries) and revised by 43 expert reviewers.

Within its pages are two new genera (Anaverticicladus from Brazil,

Capsicispora from China) and 25 species new to science representing

Anaverticicladus & Thozetella from Brazil; Boletus from U.S.A.; Capsicispora,

Corynesporopsis, Diaporthe, Diorygma, Diploschistes, Ellisembia, Erysiphe,

Gymnopus, Hemithecium, Phaeomonilia, Pseudoacrodictys, Rosellinia,

Sporidesmiella, and Stanjehughesia from China; Harmoniella from Chile;

Laboulbenia from Cameroon; Lithothelium from Bermuda; Marthamyces from

Mexico; Periconia from Thailand; and Tubulicrinis from French West Indies.

In addition to range extensions and/or new hosts for previously named

taxa in Faurelina (Brazil), Puccinia (Pakistan), Rhizocarpon (China), and

Badhamia, Didymium, Echinostelium, Perichaena, and Physarum (Mexico),

we also offer a presentation of the autoecious life cycle of Puccinia tatarinovii

(here newly lecto-/epitypified), morphological and phylogenetic analyses of

the Cladosporium causing yellow leaf spot on peppers in Cuba, and new keys

to Erysiphe on plumbaginaceous hosts, Faurelina worldwide, Hemithecium in

China, Marthamyces in America, Phaeomonilia worldwide, and Tubulicrinis

worldwide.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

8 October 2016

MY COTAXON

July-September 2016—Volume 131, pp. 491-502

http://dx.doi.org/10.5248/131.491

Periconia notabilis sp. nov. and a new record

and notes on the genus in Thailand

CHARUWAN CHUASEEHARONNACHAT , SAYANH SOMRITHIPOL

& NATTAWUT BOONYUEN

BIOTEC, National Science and Technology Development Agency (NSTDA),

113 Thailand Science Park, Thanon Phahonyothin, Tambon Khlong Nueng,

Amphoe Khlong Luang, Pathum Thani 12120, Thailand

“CORRESPONDENCE TO: charuwan.chu@biotec.or.th

ABSTRACT—Periconia notabilis is described and illustrated as a new species discovered on a

dead twig of Delonix regia in a terrestrial habitat, Pathum Thani Province, Thailand. It differs

from all other Periconia species by its pale brown to brown spherical sheath around each

conidiophore regeneration point. Periconia obscurata is reported from Thailand for the first

time, and five other Periconia spp. are described and illustrated from Thai collections.

KEY worps—asexual fungi, microfungi, decaying plants, morphology, fungal taxonomy

Introduction

Periconia (Pleosporales, Dothideomycetes) is a polyphyletic genus. Periconia

was introduced by Tode (1791) with P lichenoides as the type species. Its

systematics based on molecular phylogeny are unclear (Markovskaja &

Kacergius 2014). Periconia species are known from various geographical

locations and on different hosts, reported as herbaceous endophytes, plant

pathogens on leaves and roots, and terrestrial saprobes found in mangrove,

marine, and hypersaline ecosystems (Alias & Jones 2000; Cantrell et al.

2007; Earle 1902; Ellis 1971, 1976; El-Sharouny et al. 2009; Kohlmeyer 1977;

Markovskaja & Kacergius 2014; Rao & Rao 1964; Whitton et al. 2012).

Periconia is characterized by conidiophores that are macronematous

or sometimes micronematous, mononematous, branched or unbranched,

sometimes setose, septate, erect, pale to dark brown, smooth or rarely

492 ... Chuaseeharonnachai, Somrithipol & Boonyuen

verruculose at the apical part of conidial head and conidiogenous cells that are

monoblastic or polyblastic, ellipsoidal to spherical, discrete, formed on a stipe

or branches, sometimes the conidiogenous cells arise from the lower part of the

conidiophores. Conidia are blastocatenate, but maturing from basipetal, pale to

dark brown, aseptate, spherical, ellipsoidal to oblong, smooth or ornamented

(Ellis 1971, Markovskaja & Kacergius 2014).

Currently, the genus Periconia includes c. 40 accepted species (Kirk et al.

2008, Seifert et al. 2011).

During our continuing (2010-12) survey of saprophytic hyphomycetes

associated with plant debris and saprobic micro-fungi from several collecting

sites in Thailand, we found a new species described here as Periconia notabilis

as well as specimens of six other Periconia species: P. obscurata (a new record

for Thailand), P byssoides, P. cookei, P. echinochloae, P. lateralis, and P. sacchari.

Our descriptions and illustrations are based on morphological observations

from the natural substrate, as attempts at isolation failed for several species.

Materials & methods

Our new species, P. echinochloae, P. lateralis, and P. sacchari were collected from

decaying leaves and twigs in Pathum Thani Province; the three other Periconia species

were collected in different provinces: P byssoides and P. obscurata from Kanchanaburi

Province and P. cookei from Chiang Rai Province. Samples were taken to the laboratory

in polythene bags, rinsed in sterilized distilled water, and incubated in a moist chamber

at room temperature (~20°C) for at least 14 days. Unfortunately, attempts to isolate and

culture P notabilis using a mono-spore technique failed. Because of limited type material

and non-germinated conidia, the taxonomic description of the new species in this paper

is based only on morphological characteristics without molecular data. Fungal cultures

with successful isolation (P cookei, PR. echinochloae, P. obscurata, and P. sacchari) are

maintained in BIOTEC Culture Collection, Thailand (BCC). Voucher specimens of the

new species and other studied fungal permanent slides are deposited in the BIOTEC

Bangkok Herbarium, Thailand (BBH). Drawings were made directly from slides using a

compound microscope (Olympus BX50) equipped with a drawing tube.

Taxonomy

Periconia notabilis Chuaseehar., Somrith. & Boonyuen sp. nov. PLATE 1

MycoBank MB 815470

Differs from other Periconia species by its subspherical, pale brown to brown sheath

around the septum at the regeneration point of the conidiophore.

Type: Thailand, Pathum Thani Province, NSTDA, 14°04’33”N 100°36’21”E, on decaying

twig of Delonix regia (Hook) Raf. (Leguminosae), 14 Jul. 2011, C. Chuaseeharonnachai

(Holotype: BBH 39402).

EryMo_oey: From the Latin notabilis (noteworthy), referring to the conspicuous sheath

on the conidiophores.

Periconia notabilis sp. nov. (Thailand) ... 493

PLaTE 1. Periconia notabilis (holotype, BBH 39402). A. Conidiophores and head of conidia

with sheath (arrow). B, C. Apical part of conidiophores showing sheath around the septa at the

regeneration points (arrows), conidiogenous cells, and conidia. D. Branched chains of conidia

formed blastically. E-G. Conidia. Scale bars: A = 50 um; B, C = 25 um; D-G = 10 um.

COLONIES on natural substrate effuse, dark, hairy. Mycelium mostly immersed.

CONIDIOPHORES macronematous, mononematous, erect, 230-360 um long,

10-20 um thick at the base (x = 311 x 13.75 um; n = 10), 5-7-septate, brown

to reddish brown, thick-walled, verruculose especially below the apical

conidial heads and smooth-walled at the basal part, often branched, and with

regeneration. The primary branches are up to 15 x 7.5-10 um; the secondary

494 ... Chuaseeharonnachai, Somrithipol & Boonyuen

branches are up to 10 x 7.5 um, clavate-inflated, reddish brown, thick-walled,

verruculose. Conidiophore regeneration is terminal and percurrent, leaving a

subspherical, pale brown to brown, 30 x 22.5 um diam. sheath. CONIDIOGENOUS

CELLS monoblastic or polyblastic, discrete, subspherical to oval, smooth or

verruculose, brown to reddish brown, arising directly from the apical cells of

conidiophores, bearing simple or branched chains of conidia, 7.5-12.5 x 7.5 um

diam. ConipI<A catenate, spherical, pale brown to reddish brown, verruculose,

aseptate, 3.5-5 um diam. (x = 4.4 um; n = 50).

ComMENT: Periconia notabilis resembles P. minutissima and P nigriceps in having

conidiophores with short branches at the apical head and having spherical

verruculose conidia. However, P minutissima differs by its conidiophore

branches (at first adpressed like those of P. notabilis but later spreading) and by

PLATE 2. Periconia byssoides (BBH 39403). A, H. Conidiophores and head of conidia.

B, G. Apical parts of conidiophores, conidiogenous cells, and conidia. C. Apical portion of

conidiophore, arrow indicates a short apical cell cut off by a septum. D-F. Conidia. Scale bars:

A, H = 50 um; B-G = 25 um.

Periconia notabilis sp. nov. (Thailand) ... 495

its slightly larger (4-7 um), straw-coloured to pale brown conidia (Ellis 1971);

P. nigriceps differs by its smooth conidiophores and its larger conidia (6-8 um;

Earle 1902).

Periconia byssoides Pers., Syn. Meth. Fung.: 18 (1801) PLATE 2

COLONIES on natural substrate effuse, brown to dark brown, hairy.

MyceEL1uM mostly immersed, branched, septate, smooth, brown, 2.5 um

diam. CONIDIOPHORES macronematous, mononematous, erect, straight or

slightly flexuous, 270-350 um long, 15-17.5 um thick at the base, 7.5 um

thick at the apex, unbranched, 4-5-septate, brown, thick-walled, verruculose

immediately below the conidial heads and the lower to the basal part smooth-

walled. CONIDIOGENOUS CELLS monoblastic or polyblastic, discrete, terminals,

spherical, brown, smooth or verruculose. Conip1A solitary or catenate,

spherical, brown to reddish brown, verrucose, aseptate, 12.5-15 um diam.

SPECIMEN EXAMINED: THAILAND, KANCHANABURI PROVINCE, Suan Pa Sai Yok, on

decaying unidentified leaves, 22 Aug. 2012, C. Chuaseeharonnachai (BBH 39403).

ComMENT: Periconia byssoides resembles P. manihoticola and P. shyamala in

having unbranched conidiophores with a short apical cell cut off by a septum

and conidiogenous cells formed over apex in collar around the septum.

However, conidia of P. byssoides are much smaller than those of P manihoticola

(25-45 um) and P. shyamala (16-22 um; Ellis 1971).

Periconia cookei E.W. Mason & M.B. Ellis, Mycol. Pap. 56: 72 (1953) PLATE 3

COLONIES on natural substrate effuse, brown to dark brown, hairy.

MYcELIUM mostly immersed, branched, septate, smooth, brown, 2.5 um diam.

CONIDIOPHORES macronematous, mononematous, erect, straight or curved,

870-880 um long, 20-22.5 um thick at the base, 12.5 um thick at the apex,

unbranched, 4-septate, brown to reddish brown, thick-walled, verruculose

immediately below the conidial heads and the lower to the basal part smooth-

walled. CONIDIOGENOUS CELLS monoblastic or polyblastic, discrete, terminal,

spherical, brown to reddish brown, smooth or verruculose. Conrp1a solitary

or catenate, spherical, brown to reddish brown, verrucose, aseptate, 15-17.5

um diam.

SPECIMENS EXAMINED: THAILAND, CHIANG RAI PROVINCE, Wiang Pa Pao District,

on decaying leaves of Artocarpus altilis (Parkinson) Fosberg (Moraceae), 26 Oct. 2012, S.

Somrithipol (BBH 35459, BCC 60881; BBH 35460, BCC 60882).

ComMENT: Periconia cookei (Ellis 1971) is morphologically similar to

P. jabalpurensis and P guangdongensis in having long unbranched

conidiophores with a swollen apex. However, the verrucose conidia of P. cookei

496 ... Chuaseeharonnachai, Somrithipol & Boonyuen

A B

PLatE 3. Periconia cookei (BBH 35460). A. Conidiophore and head of conidia. B, F, G. Conidiogenous

cells and conidia. C-E. Conidia. Scale bars: A = 100 um; B, K G = 25 um; C-E = 15 um.

are larger than the echinulate conidia of P jabalpurensis (8-12 um; Ellis 1976)

and P. guangdongensis (7-11 um; Wu et al. 2015).

Periconia echinochloae (Bat.) M.B. Ellis, Demat. Hyphomyc.: 347 (1971) PLATE 4

COLONIES on natural substrate effuse, dark, hairy. MyceL1um mostly

immersed, branched, septate, smooth, brown. CONIDIOPHORES macronematous,

mononematous, erect, straight or often curved, 540-800 um long, 15-20 um

thick at the base, 7.5-10 um thick at the apex, 7-8-septate, branched; always

rising short lateral branches or formed in a group of 2-3 branches at the apex of

the stipes with secondary and tertiary branches; the primary branches are up to

Periconia notabilis sp. nov. (Thailand) ... 497

A B | F | G

o..D |

PLATE 4. Periconia echinochloae (BBH 34960). A. Conidiophore and conidial head.

B, F—G. Conidiogenous cells and conidia. C-E. Conidia. Scale bars: A = 100 um; B, F-G = 25 um;

C-E = 15 um.

30 x 6.25 um; the secondary branches are up to 17.5 x 5 um; the tertiary branches

are 12.5 x 7.5 um, brown to dark reddish brown toward the base, thick-walled,

smooth, cylindrical, brown, thick-walled, smooth. CONIDIOGENOUS CELLS

monoblastic or polyblastic, integrated, terminal, discrete, oval to cylindrical,

brown to reddish brown, smooth. Conrp1a solitary, ellipsoidal to cylindrical,

brown, verruculose, aseptate, 10.5-15 x 7.5-8.75 um diam.

SPECIMENS EXAMINED: THAILAND, PaTHuM THANI PROVINCE, NSTDA, on decaying

leaves of Saccharum officinarum L. (Poaceae), 28 Aug. 2012, S. Somrithipol (BBH 34960,

BCC 62335; BBH 34961, BCC 60844; BBH 34962, BCC 60845); Thanyaburi District,

on decaying leaves of Cymbopogon citratus (DC.) Stapf (Poaceae), 27 Aug. 2012, S.

Somrithipol (BBH 35457, BCC 60879).

ComMENT: Periconia echinochloae (Ellis 1971) is morphologically similar to

P. trachycarpicola in having branched conidiophores and ellipsoidal conidia,

but P trachycarpicola has smaller conidia (6-11x 5-7 um; Taylor & Hyde 2003).

498 ... Chuaseeharonnachai, Somrithipol & Boonyuen

PLaTE 5. Periconia lateralis (BBH 36934). A. Conidiophore, conidiogenous cells, and conidia.

B, C. Conidia. D, E. Conidiogenous cells and conidia. Scale bars: A = 50 um; B-E = 20 um.

Periconia lateralis Ellis & Everh., J. Mycol. 2: 104 (1886) PLATE 5

COLONIES on natural substrate effuse, brown to dark brown, hairy. MycELIum

mostly immersed, branched, septate, smooth, brown. CONIDIOPHORES

macronematous, mononematous, with setiform apices, erect, often curved,

300-440 um long, 17.5-20 um thick at the base, tapering to 2.5-5 um at the

apex, unbranched, 6-9 septate, brown to reddish brown toward the base, thick-

walled, smooth. CONIDIOGENOUS CELLS monoblastic or polyblastic, discrete,

directly formed in the middle part of the conidiophores, unilateral, spherical to

oval, brown to reddish brown, smooth. Conrpra solitary or catenate, spherical,

brown, verruculose, aseptate, 10.5-15 um diam.

SPECIMEN EXAMINED: THAILAND, PATHUM THANI PROVINCE, Thanyaburi District,

on decaying leaves of Brachiaria mutica (Forssk.) Stapf (Poaceae), 27 Aug. 2012, S.

Somrithipol (BBH 36934).

Periconia notabilis sp. nov. (Thailand) ... 499

CoMMENT: Morphologically, P. lateralis (Ellis 1971, Ellis & Everhart

1886) most closely resembles P fusiformis and P. pseudolateralis in having

solitary conidiophores with conidiogenous cells formed below the setiform

apices. These species are otherwise distinguished by the branching system

of conidiophores, the position of the conidiogenous cells, and conidial

morphology. Conidiophores of P pseudolateralis consist of fertile branches

spirally arranged around the stipe, and the conidia are spiny and smaller

(6-9 um; Hoog & Rao 1975). The conidiophores of P. fusiformis consist of

A C

; Pettenersegeen 4 Pa Rte . e

tee BRS asi olen : ;

ae it ati dnchich ems dad dvs bh

LE A tie,

Sie ;

lS eS

tea,

I ee

Fier

eee

Lgl tag 2

PLATE 6. Periconia obscurata (BBH 39404). A. Conidiophores and conidial head. B, F Conidiogenous

cells and conidia. C-E. Conidia. FE, G. Conidiophore, conidiogenous cells, and conidia.

Scale bars: A, G = 50 um; B, F = 25 um; C-E = 20 um.

500 ... Chuaseeharonnachai, Somrithipol & Boonyuen

fertile branches formed on all sides of the stipe, and the conidia are verrucose

and smaller (9.5-13 um; Muntafola-Cvetkovic et al. 1998).

Periconia obscurata Kirschst., Ann. Mycol. 34: 195 (1936) PLATE 6

COLONIES on natural substrate effuse, dark brown, hairy. MycELIuM mostly

immersed, branched, septate, smooth, brown, 5 um diam. CONIDIOPHORES

macronematous, mononematous, erect, straight or slightly flexuous, 200-350

um long, 12-17.5 um thick at the base, 7.5 um thick at the apex, branched;

usually rising short lateral branches at the apex, 4-6 septate, brown to dark

brown toward the base, thick-walled, smooth. The apical branches are up to

20 x 6.25-7.5 um, cylindrical, brown, thick-walled, smooth. CONIDIOGENOUS

CELLS monoblastic or polyblastic, discrete, terminal, subspherical to oval,

brown to dark brown, verruculose, 6.25-7.5 x 5-7.5 um diam. Conipiza solitary

or catenate, ellipsoidal or oval, brown, verruculose, aseptate, 6.25-10 x 5-6.25

um diam.

SPECIMEN EXAMINED: THAILAND, KANCHANABURI PROVINCE, Suan Pa Sai Yok, on

decaying unidentified twig, 22 Aug. 2012, C. Chuaseeharonnachai (BBH 39404, BCC

61896).

CoMMENT: Periconia obscurata is described here as a new record for Thailand.

The morphological characters of our specimen are consonant with the

description given by Kirschstein (1936) with the exception of length and

branching of conidiophores. The conidiophores of Kirschstein’s holotype were

smaller (200-225 x 12-16 um) and branching was not reported. This suggests

that branching and length of conidiophores are not crucial characters to

distinguish between species.

Periconia sacchari J.R. Johnst., J. Dept. Agric. Porto Rico 1: 225 (1917) PLATE 7

COLONIES on natural substrate effuse, dark brown, hairy. MycELIuM mostly

immersed, branched, septate, smooth, brown. CONIDIOPHORES macronematous,

mononematous, erect, straight or slightly flexuous, 370-400 um long,

17.5-20 um thick at the base, tapering to 6.25-10 um at the apex, branched;

producing lateral branches or formed in a group of short 2-3 branches at the

apex, 3-4-septate, brown to reddish brown toward the base, thick-walled,

smooth. Branches mainly producing secondary, and tertiary branches; the

primary branches are up to 92.5 x 5-6.25 um; the secondary branches are

10-12.5 x 5 um; the tertiary branches are 10 x 6.25 um, cylindrical, brown,

thick-walled, smooth. CONIDIOGENOUS CELLS monoblastic, discrete,

terminal, ellipsoidal, brown to reddish brown, smooth, 10-12.5 x 5-7.5 um

diam. Conip1A solitary, oblong, brown, verrucose, aseptate, 16.25-17.5 x

7.5-8.75 um diam.

Periconia notabilis sp. nov. (Thailand) ... 501

PLATE 7. Periconia sacchari (BBH 35458). A. Conidiophore and conidial head. B—D. Conidiogenous

cells and conidia. Scale bars: A = 50 um; B-D = 25 um.

SPECIMEN EXAMINED: THAILAND, PaTHUM THANI PROVINCE, Thanyaburi District,

on decaying leaves of Cymbopogon citratus, 27 Aug. 2012, S. Somrithipol (BBH 35458)

— BCC 60880 culture in BCC.

COMMENT: Periconia sacchari (Ellis 1971) differs from other Periconia species

in having oblong or cylindrical conidia. Periconia nilagirica also has elongate

conidia, but they are echinulate, with a similar range of sizes but extending to a

longer maximum (14-21 x 7-9.8 um; Subramanian 1957, 1971).

Acknowledgments

The research was financed by National Center for Genetic Engineering and

Biotechnology (BIOTEC), National Science and Technology Development Agency

(NSTDA) with the grant number P-12-01294. The authors express our gratitude to

Professor E.B. Gareth Jones at Department of Botany and Microbiology College of

502 ... Chuaseeharonnachai, Somrithipol & Boonyuen

Science King Saud University for his constructive review, critical reading and language

corrections of the draft manuscript. Dr. Natapol Pornputtapong, a Postdoctoral Associate

at Yale University, is acknowledged for his gathering supporting documents and

references. We would like to thank Prof. Dr. Akira Nakagiri and Prof. Dr. R.E Castaneda-

Ruiz for their comments and suggestions for the improvement of the manuscript, and

Dr. Shaun R. Pennycook for assistance with formatting the article.

Literature cited

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mangrove stand, Malaysia. Fungal Diversity 5: 9-21.

Cantrell SA, Hanlin RT, Emiliano A. 2007. Periconia variicolor sp. nov., a new species from Puerto

Rico. Mycologia 99(3): 482-487. http://dx.doi.org/10.3852/mycologia.99.3.482

Earle FS. 1902. Mycological studies — I. Bulletin of the New York Botanical Garden 2: 331-350.

El-Sharouny HM, Gherbawy YAMH, Abdel-Aziz FA. 2009. Fungal diversity in brackish and saline

lakes in Egypt. Nova Hedwigia 89: 437-450.

http://dx.doi.org/10.1127/0029-5035/2009/0089-0437

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew.

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew.

Ellis JB, Everhart BM. 1886. New species of fungi from various localities. Journal of Mycology 2(9):

99-104. http://dx.doi.org/10.2307/3752230

Hoog GS de, Rao V. 1975. Some new hyphomycetes. Persoonia 8(2): 207-212.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10" edn. CAB International, Wallingford Oxon, UK.

Kirschstein W. 1936. Beitrage zur Kenntnis der Ascomyceten und ihrer Nebenformen besonders

aus der Mark Brandenburg und dem Bayerischen Walde. Annales Mycologici 34(3): 180-210.

Kohlmeyer J. 1977. New genera and species of higher fungi from the deep sea (1615-5315 m).

Revue de Mycologie 41(2): 189-206.

Markovskaja S, Kacergius A. 2014. Morphological and molecular characterisation of Periconia

pseudobyssoides and closely related P byssoides. Mycological Progress 13(2): 291-302.

http://dx.doi.org/10.1007/s11557-013-0914-6

Muntanola-Cvetkovi¢ M, Hoyo P, Gémez-Bolea A. 1998. Periconia fusiformis anam. sp. nov.

Mycotaxon 68: 131-136.

Rao PR, Rao D. 1964. The genus Periconia from India. Mycopathologia et Mycologia Applicata

22(4): 285-310.

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

Fungal Biodiversity Centre, Utrecht.

Subramanian CV. 1957. Hyphomycetes - IV. Proceedings of the Indian Academy of Sciences

Section B 46: 324-335.

Subramanian CV. 1971. Hyphomycetes: an account of Indian species, except Cercosporae. Indian

Council of Agricultural Research, New Delhi. 930 p.

Taylor JE, Hyde KD. 2003. Microfungi of tropical and temperate palms. Fungal Diversity Research

Series 12. 459 p.

Tode HJ. 1791. Fungi Mecklenburgenses selecti. Fasc. II, Generum novorum appendicem,

Liineburg.

Whitton SR, McKenzie EHC, Hyde KD. 2012. Fungi associated with Pandanaceae. Fungal Diversity

Research Series 21. 457 p. http://dx.doi.org/10.1007/978-94-007-4447-9_1

Wu Y-M, Xu J-J, Kong J-H, Zhang T-Y. 2015 [“2014”]. New species of Graphium and Periconia from

China. Mycotaxon 129(2): 397-401. http://dx.doi.org/10.5248/129.397

MY COTAXON

July-September 2016— Volume 131, pp. 503-509

http://dx.doi.org/10.5248/131.503

New host records of rust fungi (Pucciniales) from Pakistan

B. ALI, Y. SOHAIL & A.S. MUMTAZ*

Department of Plant Sciences, Faculty of Biological Sciences, Quaid-i-Azam University,

Islamabad, 45320, Pakistan

*CORRESPONDENCE TO: asmumtaz@qau.edu.pk

ABSTRACT—Uredopeltis chevalieri on Grewia optiva and on G. tenax are new host records;

Broussonetia papyrifera on Cerotelium fici and Phakopsora ziziphi-vulgaris on Ziziphus

spina-christi are new host records for Pakistan; and within Pakistan P. ziziphi-vulgaris on

Ziziphus oxyphylla is a new host record for the Islamabad region. An additional collection of

Phragmidium rosae-moschatae on Rosa centifolia from Pakistan is described and illustrated.

Key worps—Hazara Division, Margalla hills, Murree, Uredinales

Introduction

Pakistan is gifted with a rich and diverse flora with approximately

6000 species of wild plants, based on which the country has been divided

into four phytogeographical regions: lIrano-Turanian (46%), Sino-

Himalayan (10%), Saharo-Sindian (9.5%), and Indian element (4.5%)

(Hamayun 2003). About 400 species of rust fungi on 350 different

host plant species have been reported in Pakistan (Ahmad et al. 1997).

Grewia tenax, Broussonetia papyrifera and Rosa centifolia are exotic plants

in Pakistan, while Grewia optiva, Ziziphus spina-christi, and Z. oxyphylla are

native plants.

Both rust fungi and their host plants are still under-explored in Pakistan. In

2014 and 2015, fieldwork was carried out in different regions of the country to

collect and investigate rust fungi and their hosts. Here we provide descriptions

and illustrations including scanning electron micrographs of four new records

of rust fungi.

Materials & methods

Different regions of Pakistan were surveyed for rust fungi in 2014-15. Collections

of the infected plant material were preserved in the herbarium of Plant Sciences

504 ... Ali, Sohail & Mumtaz

Department, Quaid-i-Azam University, Islamabad, Pakistan (ISL), and rust spores were

mounted in glycerin jelly and fixed as semi-permanent slides. The spores were measured

and photographed at 100x and 1000x magnifications under a Leitz HM-LU compound

light microscope. At least 30 spores were measured for each spore stage, including the

smallest and the largest spores found. Host plants were identified by comparing new

collections with botanical specimens held in herb. ISL. Rust fungi were identified using

the descriptions of McKenzie (1986), Walker & Shivas (2004), Sultan (2005), Berndt

& Wood (2012), Afshan et al. (2012), Fiaz et al. (2015), and Yun (2016). For scanning

electron microscopy the spores were scraped from the leaves, mounted on a stub, coated

with gold in a sputter-coater, and examined with a JSM-5910 SEM.

New records

Uredopeltis chevalieri J. Walker & R.G. Shivas, Australas. Plant Path. 33: 43

(2004) PLATE 1

= Phakopsora grewiae (Pat. & Har.) Cummins, Bull. Torrey Bot. Club 72: 206 (1945)

SPERMOGONIA, AECIA, and TELIA not observed. UREDINIA mostly

hypophyllous, scattered, about 1 mm in diam., reddish brown, subepidermal,

paraphyses clavate, fused below and free above, incurved to irregular, hyaline

to faintly yellowish, narrower at base. UREDINIOSPORES globose, subglobose to

% s DN asi x

Uredopeltis chevalieri. A. Abaxial leaf surface of Grewia optiva with brown uredinia.

B. Adaxial leaf surface of G. tenax with brown uredinia and telia. C. Urediniospores on G. tenax.

D, E. Scanning electron micrographs showing echinulate urediniospores with smooth bases.

Scale bar: C = 10 um.

New records of rust hosts for Pakistan ... 505

oval or pyriform, sometimes irregular, pale yellow to pale yellowish-brown or

rarely hyaline, (18-)20-28(-31) x (15-)17-20(-22) um in size, wall uniformly

1(-2) um thick, echinulate, spores mostly with smooth bases, 4 equatorial germ

pores.

MATERIAL EXAMINED—PAKISTAN, PunyjaB, Islamabad, Pir Sohawa, 823 m a.s.L.,

on Grewia tenax (Forssk.) Fiori, (Malvaceae), stage I], December 2014, B. Ali # BA60

(ISL-53522); on G. optiva J.R. Drumm. ex Burret, stage I], December 2014, B. Ali BA61

(ISL-29434); Islamabad, QAU, 595 m a.s.1., on G. optiva, stage II, December 2014, B. Ali

BA22 (ISL-29435).

ComMENTS— Uredopeltis chevalieri has been previously reported from Pakistan

(as Phakopsora grewiae) on Grewia asiatica (Rahber-Bhatti et al. 1988, Ahmad

et al. 1997). Grewia optiva and G. tenax are new host records for U. chevalieri

worldwide.

Cerotelium fici (Castagne) Arthur, Bull. Torrey Bot. Club 44: 509 (1917) PLATE 2

SPERMOGONIA, AECIA, and TELIA not observed. UREDINIA hypophyllous,

brown or pale brown, small, pulverulent, scattered, in groups, covered by

Cerotelium fici. A. Abaxial leaf surface of Broussonetia papyrifera with uredinia. B. Urediniospores

C, D. SEM micrographs of finely echinulate urediniospores. Scale bars: B = 8 um, C = 10 um,

D=5 um.

506 ... Ali, Sohail & Mumtaz

epidermis, surrounded by paraphyses. UREDINIOSPORES hyaline, ellipsoid,

ovoid, elongated, subglobose, 26-36 x 14-22 um in size, wall 1.5-2.5 um thick,

echinulate.

MATERIAL EXAMINED — PAKISTAN, KHYBER PAKHTUNKHWA, Hazara region,

Haripur District, 538 ma.s.1., on Broussonetia papyrifera (L.) L Hér. ex Vent. (Moraceae),

stage II, December 2014, B. Ali # BA73 (ISL-114420).

CoMMENTS — Cerotelium fici has been previously described in Pakistan on

Ficus palmata, F. carica, and F. religiosa (Ahmad 1956, Kaneko 1993, Iqbal &

Khalid 1996, Ahmad et al. 1997, Fiaz et al. 2015). Broussonetia papyrifera is

here recorded for the first time as a host of C. fici in Pakistan.

Phakopsora ziziphi-vulgaris Dietel, Ann. Mycol. 8: 469 (1910) PLATE 3

SPERMOGONIA, AECIA, and TELIA not observed. UREDINIA hypophyllous, in

irregular groups, pale brown, pulverulent. UREDINIOSPORES globose to oval or

Phakopsora ziziphi-vulgaris A. Infected leaves of Ziziphus oxyphylla. B. Urediniospores on Z. spina-

christi. C, D. SEM micrographs of echinulate urediniospores on Z. spina-christi. Scale bars: B = 10 um,

C,D=15 um.

New records of rust hosts for Pakistan ... 507

obovoid, sometimes slightly irregular, pale brown to orange brown, 17.5-27.5

x 10-17.5 um in size, wall 1.5-2.5 um thick, echinulate, smooth areas in middle

section of some spores.

MATERIAL EXAMINED — PAKISTAN, PUNJAB PROVINCE, Islamabad, Pir Sahwa, 823 m

a.s.l, on Ziziphus oxyphylla Edgew. (Rhamnaceae), stage H, December 2014, B. Ali #

BA49 (ISL-66410); Islamabad, QAU, 610 m a.s.l., on Z. spina-christi (L.) Desf., stage II,

November-December 2014, B. Ali # BAH (ISL-66411).

ComMENtTs — Phakopsora ziziphi-vulgaris has been previously reported on

Ziziphus jujuba from different regions of Pakistan (Ahmad 1956, Hasnain et

al. 1959, Iqbal & Khalid 1996, Ahmad et al. 1997) and on Z. oxyphylla from

Chakwal and Kallar Kahar (Kaneko 1993, Ahmad et al. 1997). Berndt & Wood

(2012) recorded Z. spina-christi as a host of P. ziziphi-vulgaris in Israel. Here,

Z. spina-christi is newly recorded from Pakistan as a host of P. ziziphi-vulgaris,

and Z. oxyphylla represents a new host from the Islamabad region, Punjab.

Phragmidium rosae-moschatae. A. Urediniospores with yellowish contents. B. Teliospores with

5 and 8 cells. C. Teliospore with 10 cells. Scale bars: A = 10 um, B, C = 50 um.

Phragmidium rosae-moschatae Dietel, Hedwigia 44: 132 (1905) PLaTE 4

SPERMOGONIA and AECIA not observed. UREpinIA hypophyllous, cinnamon

brown, minute, rounded, scattered. UREDINIOSPORES 22-28 x 18-22 um,

globose to subglobose, single-celled, wall hyaline to yellow brown, 2-3 um

thick, coarsely verrucose, germ pores indistinct. TeL1A hypophyllous, black,

minute, scattered. TELIOSPORES 70-130 x 24-35 um, cylindrical, rounded at

both ends, deep brown to light brown, (5—)7-8(-10)-celled, wall dark brown

to chestnut brown, 2-4 um thick, with 2 germ pores, verrucose, with an apical

papilla 7-13 um in length, hyaline. Pepice hyaline to olivaceous brown,

85-115 um, at apex 8-10 um thick, thickened to 14-25 um at base, persistent,

often with yellowish contents at base.

MATERIAL EXAMINED — PAKISTAN, KHYBER PAKHTUNKHWA, Muree, Ghora Gali,

1767 m a.s.1., on Rosa centifolia L. (Rosaceae), stages II+III, December 2014, B. Ali #

BA22 (ISL-122345).

508 ... Ali, Sohail & Mumtaz

CoMMENTS — Phragmidium rosae-moschatae has been previously reported

in Pakistan on Rosa brunonii [= R. moschata], R. lacerans, R. webbiana, and

R. centifolia from Quetta, Murree, Peshawar, Tarnab, Swat, Kaghan Valley, and

Tatu-Fairy Meadows in Pakistan (Malik & Khan 1944, Ahmad 1956, Jorstad &

Iqbal 1967, Malik et al. 1968, Ono & Kakishima 1992, Ono 1992, Kakishima

et al. 1993, Sultan 2005, Afshan et al. 2012). The number of cells in teliospores

is variable: in the species protologue, Dietel (1905) described 5-10-celled

teliospores; Pakistani teliospores have been described as (4-)7-8-celled by

Jorstad & Iqbal (1967), as 5-7(-8)-celled by Sultan (2005) and Afshan et al.

(2012), and as (5-)7-8(-10)-celled in this paper; and in a general modern

description, Yun (2016) cited 5—7(-11)-celled teliospores.

Acknowledgements

We are sincerely thankful to Dr. Amy Rossman (Department of Botany and

Plant Pathology, Oregon State University) and Dr. Merje Toome (Plant Health and

Environment Laboratory, Ministry for Primary Industries, Auckland, New Zealand) for

acting as presubmission reviewers and Dr. Shaun Pennycook (Auckland, New Zealand)

for nomenclatural review and suggestions.

Literature cited

Afshan NS, Khalid AN, Niazi AR. 2012. Some new rust fungi (Uredinales) from Fairy Meadows

Northern Areas, Pakistan. Journal of Yeast and Fungal Research 3: 65-73.

Ahmad S. 1956. Fungi of Pakistan. Biological Society of Pakistan, Lahore Monograph 1. 126 p.

Ahmad §S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Dept. Bot. Univ. Punjab, Lahore, Pakistan. 248 p.

Berndt R, Wood AR. 2012. Additions to the rust fungi of South Africa. Mycological Progress

11: 483-497. http://dx.doi.org/10.1007/s11557-011-0764-z

Dietel P. 1905. Uber die Arten der Gattung Phragmidium. Hedwigia 44: 112-132.

Fiaz M, Ahmad H, Afshan NS, Khalid AN. 2015. Some new records of Uredinales from Khyber

Pakhtunkhwa, Pakistan. Mycotaxon 130: 569-575. http://dx.doi.org/10.5248/130.569

Hamayun M. 2003. Ethnobotanical studies of some useful shrubs and trees of District Buner,

NWEP, Pakistan. Journal of Ethnobotanical Leaflets, Southern Illinois University, Carbondale

IL, USA. 17 p. opensiuc.lib.siu.edu/cgi/viewcontent.cgi?article=1563&context=ebl

Hasnain SZ, Khan A, Zaidi AJ. 1959. Rusts and smuts of Karachi. Dept. Bot., Univ. Karachi,

Monograph 2. 33 p.

Iqbal SH, Khalid AN. 1996. Material for the fungus flora of Pakistan. II. An updated check list of

rust fungi (Uredinales) of Pakistan. Sultania 1: 39-67.

Jorstad I, Iqbal SH. 1967. Uredinales from West Pakistan. Nytt Magasin for Botanikk 14: 31-38.

Kakishima M, Izumi O, Ono Y. 1993. Rust fungi (Uredinales) of Pakistan collected in 1991.

Cryptogamic Flora of Pakistan 2: 169-179.

Kaneko S. 1993. Parasitic Fungi on Woody Plants from Pakistan Himalaya. Cryptogamic Flora of

Pakistan 2: 149-168.

Malik SA, Javaid MT, Ahmad M. 1968. Uredinales of Quetta-Kalat region of Pakistan. Biologia 14:

37-46.

New records of rust hosts for Pakistan ... 509

Malik SA, Khan MA. 1944. Parasitic fungi of the North-West Frontier Province. Indian Journal of

Agricultural Sciences 13: 522-527.

McKenzie EHC, 1986. New plant disease record in New Zealand: Fig rust (Cerotelium fici) on Ficus

carica. New Zealand Journal of Agricultural Research 29: 707-710.

http://dx.10.1080/00288233.1986.10430467

Ono Y, Kakishima M. 1992. Uredinales collected in the Swat Valley, Pakistan. Cryptogamic Flora

of Pakistan 1: 197-216.

Ono Y. 1992. Uredinales collected in the Kaghan Valley, Pakistan. Cryptogamic Flora of Pakistan

1: 217-240.

Rahber-Bhatti MH, Memon MI, Shahani NM, Khuhro MA. 1988. Laboratory evaluation of some

plants against rust. Pakistan Journal of Agricultural Research 9: 274-277.

Sultan MA. 2005. Taxonomic study of rust flora of Northern Areas of Pakistan. Ph.D. Thesis,

University of the Punjab. Lahore, Pakistan.

Walker J, Shivas RG. 2004. Uredopeltis chevalieri sp. nov., the rust of Grewia (Tiliaceae) formerly

known as Phakopsora (or Dasturella) grewiae, its first record in Australia and a summary of the

known rusts of Grewia. Australasian Plant Pathology 33: 41-47.

Yun HY. 2016. Musk rose rust - Phragmidium rosae-moschatae. Systematic Mycology and

Microbiology, ARS, USDA - Invasive and Emerging Fungal Pathogens - Diagnostic Fact

Sheets. http://nt.ars-grin.gov/sbmlweb/fungi/diagnosticfactsheets.cfm [Retrieved 30 January

2016].

510... Ali, Sohail & Mumtaz

MY COTAXON

July-September 2016—Volume 131, pp. 511-520

http://dx.doi.org/10.5248/131.511

New records of myxomycetes from México

Marcos LIZARRAGA’, GABRIEL MORENO”? & MARTIN ESQUEDA?

' Dpto. Ciencias Quimico Bioldgicas, Instituto de Ciencias Biomédicas,

Univ. Autonoma de Ciudad Juarez, Anillo Envolvente Pronaf y Estocolmo s/n,

Ciudad Juarez, Chihuahua 32300, México

? Dpto. Ciencias de la Vida, Edificio de Biologia, Univ. Alcala,

28805 Madrid, Spain

> Centro de Investigacion en Alimentacion y Desarrollo,

A.C. Apartado Postal 1735, Hermosillo, Sonora, México

* CORRESPONDENCE TO: gabriel. moreno@uah.es

ABSTRACT—Badhamia rhytidosperma, Didymium synsporon, Echinostelium coelocephalum,

Perichaena polygonospora, and Physarum synsporum are described in both macroscopic and

microscopic detail and represent new records from Mexico.

Key worps—Amoebozoa, chorology, myxobiota, slime moulds, taxonomy

Introduction

The present paper represents a contribution to a program of study on

myxomycetes in Mexico that began 19 years ago and from which several

publications have been generated. Most of the latter have been compiled into

Mexicos myxomycete inventory by Moreno et al. (2007). In this paper, we

present new records of several species that are poorly known worldwide and

thus are scarcely referenced in the specialized literature regarding this group

of organisms.

Materials & methods

The studied material was obtained from bark samples collected from living trees in

several Mexican states (Chihuahua, Mexico City, and Sinaloa) and placed in more than

200 moist chambers. Only species identified as new records for the Mexican myxobiota

are reported here.

512 ... Lizarraga, Moreno & Esqueda

Spores (including surface structures such as spines or warts) were measured

with an oil immersion lens. Light microscopy (LM) was carried out with a Nikon

microscope equipped with an automatic photographic system. SEM micrographs were

produced with a Zeiss DSM-950 microscope. For ultramicroscopic studies, material was

rehydrated in concentrated (28-30%) ammonium hydroxide for 30 min, dehydrated in

aqueous ethanol (70%) for 30 min, fixed for 2 h in pure ethylene glycol dimethyl ether (=

1,2-dimethoxymethane), and finally immersed in pure acetone for at least 2 h. This was

followed by critical point drying and sputtering with gold-palladium. This technique

allows the use of very little material (ie., part of a single sporocarp or sometimes no

more than a few spores).

Specimens are conserved in the herbarium of the Life Sciences Department (Botany),

University of Alcala, Madrid, Spain (AH) with a duplicate of some specimens deposited

in the Fungi Collection of Biomedical Sciences Institute, Autonomous University of

Ciudad Juarez, México (UACJ). Permanent slides were prepared with Hoyer’s liquid for

microscopic study.

Taxonomy

Badhamia rhytidosperma H.W. Keller & Schokn., Mycologia 81: 783 (1989) Fras

1-4

FRUCTIFICATIONS sessile white plasmodiocarps. SPOROTHECA globose,

subglobose to lobular with gyrose-confluent irregular lobes of 0.4-0.6 x

0.7-1.0 mm. PERIpIuM double, calcareous external layer smooth to slightly

rough, internal layer membranous with irregular dehiscence. HYPOTHALLUS

inconspicuous. CAPILLITIUM physaroid or more rarely badhamioid, formed

by joined filaments through elongated to irregular calcareous nodules. When

the capillitium is badhamioid a pseudocolumella appears at the center of

sporotheca, which is thick, irregular and calcareous. Spores (10-)12-14(-15)

um diam., black in mass and violaceous under LM, globose to subglobose,

surface smooth over one half of the spore but markedly reticulate over the

other half. Under SEM a reticulate ornamentation is confirmed in one side

and smooth on the opposite side with a distinct ridge or additional lines as

indicated by Keller & Schoknecht (1989).

SPECIMEN EXAMINED: MEXICO, CurHuanua: Biomedical Sciences Institute gardens,

Autonomous University of Ciudad Juarez, on bark of Morus nigra L., 5 February

Fics 1-13. Badhamia rhytidosperma (AH 45861). 1, 2. Plasmodiocarps. 3. Spore showing the

reticulate wall as observed by SEM. 4. Spore showing a smooth area with adjacent ridge by SEM.

Didymium synsporon (AH 45866). 5. Sporocarp. 6. Plasmodiocarp. 7. Spore with bacula outside

and smooth inside by SEM. 8. Details of bacula with a verrucose apex and reticulate base by SEM.

9. Spore cluster by SEM. 10. Detail inside a spore cluster by SEM. Echinostelium coelocephalum

(AH 45867). 11. Spore-like body with thick articulate surfaces. 12, 13. Spores with thick articulate

surfaces and regular thickness at contact points between spores. Scale bars: 1, 2, 5, 6 = 0.5 mm;

3, 4,7 =2 mm; 8 = 1 um; 9, 10 = 5 um.

513

Five myxomycetes new to Mexico ...

514... Lizarraga, Moreno & Esqueda

2000, leg. M. Lizarraga; obtained from moist chamber 21 February 2000 (UACJ 190,

AH 45861).

ADDITIONAL SPECIMEN EXAMINED: Badhamia spinispora: MEXICO, Sonora:

El Pinacate y Gran Desierto de Altar biosphere reserve, on bark of Parkinsonia sp.,

18 February 2012, leg. M. Lizarraga; obtained from moist chamber 16 March 2012

(AH 45862).

HABITAT & DISTRIBUTION—Badhamia rhytidosperma has been described

previously from two field collections on alfalfa remnants from the type

locality (Kansas, USA) and from moist chamber cultures prepared with cow

dung (Keller & Schoknecht 1989). The specimen (AH 45862) from Sonora,

Mexico, recorded originally as B. rhytidosperma by Esqueda et al. (2013) is

here redetermined as B. spinispora. Consequently our Chihuahua specimen

represents the first authentic Mexican record of B. rhytidosperma.

OBSERVATIONS—BADHAMIA RHYTIDOSPERMA is morphologically similar

to Badhamia spinispora (Eliasson & N. Lundq.) H.W. Keller & Schokn.,

B. verrucospora G. Moreno et al., and Physarum apiculosporum Hark. from

which it is clearly distinguished by its spore ornamentation. Spores are spiny

in B. spinispora, verrucose in B. verrucospora, and smooth in P apiculosporum

(Moreno et al. 2011), whereas Badhamia rhytidosperma has globose spores in

which half of the spore is smooth and the other is reticulate. On its surface,

a crested conspicuous line is observed at the site where the spore germinates

(Fic 4). Based on the literature, only the type material described by Keller &

Schoknecht (1989) was previously known.

Didymium synsporon T.E. Brooks & H.W. Keller, Mycologia 65: 287 (1973)

Fies 5-10

FRUCTIFICATIONS sessile, sporocarpic, or (more rarely) plasmodiocarpic,

ranging from pulvinate to flat and occasionally umbilicate, 0.3-1 x 0.1-0.8

mm, whitish to grey-white. PERrp1uM formed by two joined layers, the external

layer crust-like (similar to an eggshell) and the inner layer membranous and

translucent. A columella was not observed. CapiLLiTIuM is formed by rigid

and parallel filaments arising from the sporocarp base towards its apex; the

ends are dichotomous or bifurcate with few transversal junctions, dark to

hyaline at apex of 1-3(-4) um diam. and few enlargements. Spores dark in

mass and violaceous under LM, in clusters of 8-20, 10-12 um diam., pyriform

to subglobose, conspicuous ornamentation formed by fused warts that provide

a reticular to subreticular aspect, the ornamentation becoming less dense

(formed of small sparse warts) toward the base where attached to other spores.

Under SEM, the spore ornamentation is formed by bacula with a verrucose

apex and reticulate base.

Five myxomycetes new to Mexico ... 515

SPECIMENS EXAMINED: MEXICO, CuInuAHUA: Cumbres de Majalca, on bark of

Juniperus sp., 22 September 2003, leg. M. Lizarraga; obtained from moist chamber 29

September 2003 (UACJ 493); obtained from moist chamber 30 September 2003 (UAC]

466); obtained from moist chamber 1 October 2003 (UACJ 504, AH 45865); obtained

from moist chamber 8 October 2003 (UACJ 470); 8 August 2014, leg. M. Lizarraga;

obtained from moist chamber 14 August 2014 (AH 45866).

HABITAT & DISTRIBUTION—Didymium synsporon was first described in the

United States from bark sampled from living Juniperus virginiana L., cultured

in moist chambers, and there are limited records in the United States (Arkansas,

Kansas, Kentucky, and Tennessee; Keller & Brooks 1973). The species was also

observed from a collection made on Opuntia sp. from Atacama, Chile (Lado et

al. 2007).

OBSERVATIONS— This species is distinguished by its crust-like external peridium,

spore clusters, and a capillitium formed by rigid parallel filaments rising from

the sporocarp base towards the apex. It is the only known Didymium species

with clustered spores.

Echinostelium coelocephalum T.E. Brooks & H.W. Keller, Mycologia 68: 1212

(1977) [“1976"]. Fries 11-13

SPOROCARPS isolated to gregarious, stipitate, 45-60 um total height, whitish

to slightly yellow. Stipe 20-30 um long, cylindrical and expanded towards the

base to 4-6 um diam., becoming thinner towards the apex of 1-1.5 um diam.

SPOROTHECA globose to subglobose, 20-30 um diam., white to slightly yellow.

CAPILLITIUM absent. PERIDIUM persistent, similar to a thick collar at columella

base of 5-8 um diam. Sporss hyaline in mass and whitish to yellowish under

LM, globose to subglobose, 7-10 um diam., faintly verrucose with thick

articulate surfaces and regular thickness, c. 1 um thick at contact point between

spores.

SPECIMEN EXAMINED: MEXICO, CuInvuAHUA: Cumbres de Majalca, on bark of Quercus

sp., 15 November 2003, leg. M. Lizarraga & G. Mendoza; obtained from moist chamber

6 May 2004 (UACJ 605, AH 45867).

HABITAT & DISTRIBUTION—Echinostelium coelocephalum was described

from Arkansas in the United States from bark samples collected in different

American states and cultured in moist chambers (Keller & Brooks 1977). It has

since been reported from Argentina (Lado et al. 2014), Australia (Davison et al.

2008), Belize (Lado et al. 2008), France (Mitchell et al. 1984), New Caledonia

(Kylin et al. 2013), the Republic of South Africa (Ndiritu et al. 2009), Spain

(Pando 1989), U.S.A. (Haskins et al. 2000), and Kazakhstan, Ukraine, Tunisia,

and Turkey (Discover Life 2015). It is possibly an extensively distributed species

that has been overlooked because of its small size and resemblance to other

similar species.

516 ... Lizarraga, Moreno & Esqueda

OBSERVATIONS—Characterized by its small size and articulate spores with

evenly thick walls, E. coelocephalum may be confused with E. colliculosum K.D.

Whitney & H.W. Keller, which differs by its larger fructifications (70-150 um)

and spores with narrow articulations. Haskins & McGuinness (1986) examined

the microwarts on the spore surface of E. coelocephalum under the SEM, and

Haskins et al. (2000) investigated heterothallic mating systems in this species.

Perichaena polygonospora Novozh., Zeml., Schnittler & S.L. Stephenson, Mycologia

100: 819 (2008) Fries 14-19

SPOROCARPS sessile, isolated to gregarious. SPOROTHECA very small,

0.1-0.15 mm diam., globose to subglobose, yellow-orange to dark yellow.

PERIDIUM simple and membranous, frequently appearing as externally

ornamented with dark-brown small protrusions or warts homogenously

distributed on its surface and composed of granulated material; inner surface

smooth under LM. Dehiscence irregular. CAPILLITIUM absent. SPORES

yellowish in mass and bright yellow under LM, (10-)11-12(-13) um diam.,

globose to subglobose, polyhedral, verrucose. The polyhedral morphology

of the verrucose spores and its well-defined smooth angles were confirmed

under SEM. Spore ornamentation consists of fused bacula that form pyramidal

structures of different sizes and thickness.

SPECIMENS EXAMINED: MEXICO, Mexico City: Bosque de Chapultepec, on bark

of Taxodium mucronatum Ten., 30 March 2015, leg. M. Lizarraga; obtained from

moist chamber 16 May 2015 (AH 45868); obtained from moist chamber 3 June 2015

(AH 45869); obtained from moist chamber 11 June 2015 (AH 45870).

HABITAT & DISTRIBUTION—Perichaena polygonospora fruits on litter and small

twigs of shrubs but also on weathered herbivore dung (Novozhilov et al. 2008).

Studied specimens were collected from New Mexico (United States) on elk

dung (Cervus canadensis) (Mitchell & Moreno 2013) from Mexico City on bark

of Taxodium mucronatum. This species was originally described in Russia and

Kazakhstan by Novozhilov et al. (2008) and recorded and described later in the

United States by Mitchell & Moreno (2013).

OBSERVATIONS—Perichaena polygonospora is characterized by its minute

(0.1-0.15 mm diam) globose sporotheca, generally with dark brown

protrusions attached to the peridium, irregular dehiscence, polyhedral spores,

and lack of a capillitium. A similar species, Licea eremophila D. Wrigley et al.,

also has similarly sized [(9-)10-12(-13) um diam] polyhedral spores but can

Figs 14-25. Perichaena polygonospora (AH 45870, except for Fics 14, 15). 14. Gregarious

sporocarps (AH 45557). 15. Sporocarp detail (AH 45557). 16. Peridium with small warts dark-

brown. 17, 18. Polyhedral spores by SEM. 19. Detail of spore ornamentation by SEM. Physarum

synsporum (AH 45557, except for Fic. 20). 20. Fructifications sporangiate to plasmodiocarpic

Five myxomycetes new to Mexico ... 517

(AH 45872). 21. Branched sporotheca. 22. Spore with bacula on the outside by SEM. 23. Spore with

bacula on the outside and smooth on the inner surface by SEM. 24. Spore cluster by SEM. 25. Detail

of an internal spore cluster by SEM. Scale bars: 14-16 = 100 um; 17, 18, 22, 23 = 2 um; 19 = 1 um;

20, 21 = 0.5 mm; 24, 25 = 5 um.

518 ... Lizarraga, Moreno & Esqueda

be distinguished by its stipitate sporotheca, a peridium formed by plates, and

its habitat on dead leaf bases of the bromeliad Puya spp. and dead remains of

the cacti Trichocereus sp. and Miqueliopuntia miquelii (Monv.) F. Ritter from

Argentina and Chile (Wrigley de Basanta et al. 2010).

Perichaena polygonospora belongs to the Perichaena group that lacks a

capillitium and includes such species as P. pachyderma D.W. Mitchell et al.,

which also fruits on herbivore dung (deer, elk, and cow) but differs by its

globose, verrucose, non-polyhedral spores (10-11(-12) um diam.; Mitchell et

al. 2011).

Physarum synsporum S.L. Stephenson & Nann.-Bremek., Proc. Kon. Ned. Akad.

Wetensch., C. 93:193 (1990) Fics 20-25

FRUCTIFICATIONS solitary to gregarious, sporangiate to plasmodiocarpic,

sessile. SPOROTHECA subglobose, flat and somewhat branched, 0.3-1.5 x

0.3-0.5 mm, yellowish to grey-yellowish. PERIDIUM simple and calcareous,

superficially granulated due to calcium carbonate nodules lifting the peridium.

Dehiscence irregular. CAPILLITIUM formed by a calcareous nodule network

that ranges from subglobose to fusiform, whitish and joined by small hyaline

filaments. Spores dark brown in mass and violaceous brown under LM,

10-12 um diam., in clusters consisting of 4-8 pyriform, turbinate to subglobose

spores that are easily dispersed; the surface superficially verrucose on the spore

base but becoming less dense (small sparse warts) closer to the attachment

zone with the next spore. SEM confirms the turbinate morphology and reveals

an ornamentation composed of abundant bacula outside but very scarce and

smaller to almost absent inside. No plasmodium was observed.

SPECIMENS EXAMINED: MEXICO, SINALOA: Culiacan, Colonia Guadalupe Victoria, on

bark of Mangifera indica L., 20 February 2013, leg. M. Lizarraga; obtained from moist

chamber 5 September 2014 (AH 45871); obtained from moist chamber 1 October 2014

(AH 45872).

HABITAT & DISTRIBUTION—Physarum synsporum was described in the United

States based on material appearing on samples of Juniperus virginiana ina moist

chamber. Subsequently, it has been reported only on shrub twigs in Argentina

(Lado et al. 2011).

OBSERVATIONS—The diagnostic characters of P synsporum are the

plasmodiocarpic to sporocarpic fructifications with a yellowish and strongly

granulated peridium as well as clustered spores, which are easily dispersed,

allowing the observation of its pyriform or turbinate shapes with their marked

ornamentation at the external portion of the spore.

Physarum lakhanpalii Nann.-Bremek. & Y. Yamam., a species difficult to

characterize, is distinguished by its plasmodiocarpic yellowish fructifications,

Five myxomycetes new to Mexico ... 519

yellow carbonate calcium nodules, and subglobose (11-12.5 um diam.) densely

verrucose spores in clusters of 4-6 (Nannenga-Bremekamp & Yamamoto 1987);

because the protologue does not indicate the distribution of the verrucae, we

assume that they are homogeneously distributed all along the edge of the spore.

Shortly after the publication of P lakhanpalii, a second species with

clustered spores (P synsporum) was described that is characterized by dark-

brown plasmodiocarpic fructifications, whitish calcium carbonate nodules,

and clusters of 3-8 subglobose to turbinate spores (12-13 um diam), smooth or

displaying few small warts on the inside (Stephenson & Nannenga-Bremekamp

1990). These authors separate P synsporum from P lakhanpalii by its capillitium

with whitish nodules and its turbinate spore clusters with warts obvious only

on the external area.

The specimens from Mexico correlate well microscopically with the

description of P. synsporum, although the fructification has a yellowish color

instead of dark-brown as originally described. According to our experience, as

this species was obtained from a moist chamber, it probably did not optimally

develop the peridium pigmentation. Physarum decipiens M.A. Curtis is very

similar to P synsporum, differing only in the absence of clustered spores.

Acknowledgements

We wish to express our gratitude to Mr. A. Priego and Mr. J. A. Pérez of the Electron

Microscopy Service of the University of Alcala de Henares for their invaluable help with

the SEM. We also thank L. Monje and A. Pueblas of the Department of Drawing and

Scientific Photography at the Alcala University for their help in the digital preparation

of photographs and to Dr. J. Rejos, curator of the AH herbarium for his assistance with

the specimens examined in the present study. We want to express our gratitude to

S.L. Stephenson and J.R. Deschamps for reviewing the manuscript. M. Lizarraga extends

thanks to Dr. Alejandro Martinez of Dpto. Ciencias Quimico Bioldégicas, Universidad

Autonoma de Ciudad Juarez, for economic support.

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MYCOTAXON

July-September 2016—Volume 131, pp. 521-526

http://dx.doi.org/10.5248/131.521

Marthamyces coronadoae sp. nov. in a

Fagus grandifolia subsp. mexicana forest

from Hidalgo State, México

TANIA RAYMUNDO’, RICARDO VALENZUELA™ & MARTIN ESQUEDA?

‘Escuela Nacional de Ciencias Bioldgicas, Instituto Politécnico Nacional,

Plan de Ayala y Carpio s/n Col. Santo Tomas, México, D.F. 11340, México

?Centro de Investigacién en Alimentacion y Desarrollo,

A.C. Apartado Postal 1735, Hermosillo, Sonora 83000, México

" CORRESPONDENCE TO: rvalenzg@ipn.mx

AsBstTRAcT—Marthamyces coronadoae is described as a new species from a relict forest of

Fagus grandifolia subsp. mexicana in México. The specimens were collected on fallen and

dead Fagus leaves in La Mojonera (in Zacualtipan municipality, Hidalgo State, México). This

species is characterized by an apothecium opening by 4-5 pruinose and brownish beige teeth,

a pale grey to pale yellow hymenial surface, and ascospores that are curved or coiled when

released. A key to species of Marthamyces that grow in America is presented.

Key worps—Leotiomycetes, Marthamycetaceae, propoloid fungi, taxonomy

Introduction

Minter (2003) established Marthamyces (with M. emarginatus (Cooke &

Massee) Minter as type species) to accommodate taxa with filiform ascospores

placed in Propolis sensu Sherwood (1977). The genus includes species that grow

on abaxial and adaxial surfaces of fallen and dead plant leaves throughout the

world, mainly in tropical and subtropical regions (Johnston 2006). Marthamyces

is characterized by its apothecial ascomata that are subepidermal, orbicular to

polygonal, resembling small pustules, erumpent from the substrate surface,

not associated with bleaching of the surrounding substrate, and lacking zone

lines (Minter 2003, Johnston 2006). The genus is characterized macroscopically

by apothecia that are deeply immersed within the host tissue and a hymenial

surface that often appears pruinose due to the presence of small crystals

522 ... Raymundo, Valenzuela & Esqueda

amongst the paraphyses (Johnston 2006). Microscopically, the ascospores

are filiform, 0-3-septate, and with apical and basal gelatinous caps, and the

paraphyses have short branches near apex, tapering or swelling, tangled or with

several short, finger-like projections, either often intermixed with crystals, or

coated with crystals and with the tips of the paraphyses sometimes embedded

in brownish gel (Sherwood 1977, Johnston 2006). The genus was formerly

placed in Rhytismataceae, Rhytismatales, but recent phylogenetic studies in

Leotiomycetes placed Marthamyces and Propolis in a clade separate from the

Rhytismatales (Hustad & Miller 2011, Lantz et al. 2011). In 2015, Baral et al.

established a new family Marthamycetaceae to accommodate the propoloid

fungi (Baral 2015) and included Cyclaneusma, Marthamyces, Mellitiosporiella,

Mellitiosporium, Naemacyclus, Phragmiticola, Propolina, and Propolis.

In México, only Marthamyces quadrifidus (Lév.) Minter has been recorded

(specimen CU-ME-317, as Propolis quadrifida, from Oaxaca state; Sherwood

1977). In 2012, on a field survey of the relict forest of Fagus grandifolia subsp.

mexicana (Martinez) A.E. Murray in La Mojonera (municipality Zacualtipan,

Hidalgo State), México, several fallen leaves were collected that bore apothecia

of the fungus here described as a new species of Marthamyces.

Materials & methods

The La Mojonera relict forest of Fagus grandifolia subsp. mexicana (Fagaceae) covers

45 ha at 2597 m asl. Its “montane cloud forest” vegetation comprises three arborous

strata: higher (20-40 m), medium (8-20 m), and lower (2-8 m), and the higher stratum

is dominated by Fagus, associated with Magnolia schiedeana, Clethra macrophylla,

several Quercus species, Pinus greggii, and P patula (Alcantara & Luna-Vega 2001,

Williams-Linera et al. 2003, Godinez-Ibarra et al. 2007). Our specimens are deposited

in the Herbarium, Instituto Politécnico Nacional, Mexico City, México (ENCB).

Longitude and latitude coordinates were obtained with GPS eTrex (Garmin). Colors

are coded according to Kornerup & Wanscher (1978). Morphological examinations

were conducted as outlined by Sherwood (1977) and Johnston (2006). Measurements

of anatomical characters were taken from rehydrated tissues in 5% aqueous KOH

and Melzer’s reagent. The macro-photos were taken with a Nikon D7000 and the

micrographs with a Sony DSC-WX350. The meanings of some terms are based on Ulloa

& Hanlin (2012) dictionary.

Taxonomy

Marthamyces coronadoae Raymundo, R. Valenz. & Esqueda, sp.nov. — Fics 1-19

MycoBAnk MB 815275

Differs from Marthamyces quadrifidus by its dark apothecial teeth, curved to coiled

ascospores when released, and paraphyses with several short, finger-like projections.

Type: México, Hidalgo, municipality of Zacualtipan, Ejido “La Mojonera’, 28°06’56”N

111°01’35”W, alt. 2550 m, 17 July 2012, T. Raymundo 4324 (Holotype, ENCB).

Marthamyces coronadoae sp. nov. (Mexico) ... 523

Fics 1-6: Marthamyces coronadoae (holotype, ENCB Raymundo 4324). 1 Abaxial side of leaf.

2. Pustules erumpent on surface of leaf. 3, 5. Apothecia opening in teeth. 4, 6. Adaxial side of leaf.

Scale bars: 1 = 10 mm; 2, 4= 3 mm; 3 = 500 um; 5 = 1mm.6=5 mm.

524 ... Raymundo, Valenzuela & Esqueda

Erymotoey: In honor of Sonoran researcher Ecol. Martha Coronado, a pioneer in

fungal ecology in Sonora State.

APOTHECIA 0.4-0.8 mm diam., on abaxial and adaxial surfaces of dead

coriaceous leaves, subepidermal, resembling small pustules, visible initially as

round black patches on leaf surface that raise as ascomata mature, erumpent

from substrate surface, not associated with bleaching of host tissue, without zone

lines, orbicular to polygonal, some rhomboid to pentagonal, others irregular

in shape, superficial layer black in young apothecia, covering the hymenium,

opening by 4-5 prominent, irregular, pruinose and brownish beige (6E3) teeth.

HYMENIUM somewhat depressed below substrate level, pruinose, pale grey

(1B1) to pale yellow (3A3) or pastel yellow (3A4) when fresh. Perimeter line

absent. SUBHYMENIUM 10-15 um thick, comprising small angular to globose

cells, 1-2 um diam., with hyaline walls. PARAPHYSES <100 um long and 2 um

diam. (swelling slightly near apex <3 um diam.) with several short, finger-like

projections, at the apex coated with KOH-soluble crystals. Asc 70-90 x 6-8

um, cylindric, with a foot-like base and a poorly developed basal stalk, tapering

slightly at rounded apex, wall undifferentiated at apex, 8-spored. ASCOSPORES

slightly interwoven to irregularly arranged, or subparallel in the ascus, 60-74

(-80) x 1.6-2.4 um, filiform, curved to coiled on release, hyaline in KOH, thin-

walled, 0-1-septate, apical and basal gelatinous caps globose, 2.5-3 um diam.,

gelatinous caps soluble in KOH.

ECOLOGY & DISTRIBUTION—Found growing on fallen and dead leaves of

Fagus grandifolia subsp. mexicana in montane cloud forest; known only from

the type locality.

ComMMENTS—Marthamyces coronadoae is characterized by its orbicular to

polygonal apothecia, the young apothecia erumpent through from a black

covering layer, opening by 4-5 prominent, irregular and brownish beige teeth,

its ascospores curved to coiled on release, 0-1-septate. Marthamyces quadrifidus

is a similar species to M. coronadoae, but has 1-3-septate ascospores, more

or less straight on release (Sherwood 1977, Cabarroi-Hernandez et al. 2014)

and it grows on several tropical and subtropical hosts (Clusia, Eucalyptus,

Lepidosperma, Mangifera, Myrtus, Rapanea, etc.). Marthamyces quercifolius

(Cooke & Ellis) Minter on Quercus and Marthamyces phacidioides (Fr.) Minter

on Ericaceae and Myricaceae are also similar to M. coronadoae, but they differ

by their 3-septate ascospores and their host preferences (Sherwood 1977).

Key to the species of Marthamyces in America

1. Ascospores 0-1-septate, apothecial teeth dark ........... 0. cece eee eee eee 2

1. Ascospores 3-septate, apothecial teeth pale .............. eee eee eee eee eee 3

Marthamyces coronadoae sp. nov. (Mexico) ... 525

a Ah

RSS PT a Tey

AUNT,

Fics 7-19: Marthamyces coronadoae (holotype, ENCB Raymundo 4324). 7, 8. Apothecium in

transversal cut. 9. Crystals cover hymenial surface. 10. Paraphyses and asci on hymenial surface.

11. Apex of paraphyses with several short, finger-like projections. 12-14. Asci. 15-19. Ascospores.

Scale bars: 7, 8 = 100 um; 9, 10 = 10 um; 11, 19 = 3 pm; 12, 15-18 = 5 um. 13-14 = 8 um.

526 ... Raymundo, Valenzuela & Esqueda

2. On Eucalyptus, apothecia opening by 3-5 pruinose and black teeth,

hymenial surface pale grey,

ascospores straight or slightly curved on release.............. M. emarginatus

2. On Fagus, apothecia opening by 4-5 pruinose and brownish beige teeth,

hymenial surface pale grey to pale yellow,

ascospores curved to coiled on release ........... 0... eee eee M. coronadoae

3. Tropical species, on Clusia, Conocarpus, Mangifera, Myrtus, Rapanea,

and many other hosts; hymenial surface pale grey ............. M. quadrifidus

3. Temperate species, on Quercus, Ericaceae, and Myricaceae;

Miperin eta tal Sut ae eye WO a aig eae wile cue Pues ate WA ets vat ios ate ters aay Gates dares 4

4. On Quercus, apothecia amphigenous;

ascospores 50-65 x 1.5 um, straight on release ................ M. quercifolius

4. On Ericaceae and Myricaceae, apothecia hypophyllous;

ascospores 60-75 x 1.5-2 um, straight on release............. M. phacidioides

Acknowledgments

We wish to express our gratitude to Dr. Peter R. Johnston and Dr. José G. Marmolejo

for reviewing the manuscript and their useful comments. We also express our gratitude

to SEMARNAT—CONACYT (Grant 2002-C01-0409) for their financial support.

Aldo Gutiérrez (CIAD) kindly prepared the manuscript and plates. R. Valenzuela and

T. Raymundo thank COFAA and IPN for the financial support for their research in the

project SIP-20150540, SIP-20151530, SIP-20161164, and SIP-20161166.

Literature cited

Alcantara O, Luna-Vega I. 2001. Analisis floristico de dos areas con Bosque Mesofilo de Montafia

en el Estado de Hidalgo, México: Eloxotlan y Tlahuelompa. Acta Botanica Mexicana 54: 51-87.

Baral HO. 2015. Nomenclatural novelties : H.O. Baral. Index Fungorum 225: 1-3.

Cabarroi-Hernandez M, Johnston PR, Minter DW. 2014. Rhytismataceae (Ascomycota) in Cuba.

Willdenowia 44: 65-75. http://dx.doi.org/10.3372/wi.44.44110

Godinez-Ibarra O, Pérez A, Lépez-Mata G, Garcia-Moya L, Valdez-Hernandez E, de los Santos-

Posadas Jl, Trinidad-Santos A. 2007. Lluvia de semillas y emergencia de plantulas de

Fagus grandifolia subsp. mexicana en La Mojonera, Hidalgo, México. Revista Mexicana de

Biodiversidad 78: 117-128.

Hustad VP, Miller AM. 2011. Phylogenetic placement of four genera within the Leotiomycetes

(Ascomycota). North American Fungi 6(9): 1-13. http://dx.doi.org/10.2509/naf2011.006.009

Johnston PR. 2006. Rhytismatales of Australia: the genus Marthamyces. Australian Systematic

Botany 19: 135-146. http://dx.doi.org/10.1071/SB05010

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3th ed. Eyre Methuen, London.

Lantz H, Johnston PR, Park D, Minter DW. 2011. Molecular phylogeny reveals a core clade of

Rhytismatales. Mycologia 103: 57-74. http://dx.doi.org/10.3852/10-060

Minter DW. 2003. Propolis and Marthamyces gen. nov. (Rhytismatales). Mycotaxon 87: 43-52.

Sherwood MA. 1977: Taxonomic studies in the Phacidiales: Propolis and Propomyces. Mycotaxon

5: 320-330.

Ulloa M, Hanlin RT. 2012. Illustrated dictionary of mycology. 2nd ed. APS Press, St. Paul,

Minnesota, USA.

Williams-Linera G, Rowden A, Newton AC. 2003. Distribution and stand characteristics of relict

population of Mexican beech (Fagus grandifolia var. mexicana). Biological Conservation

109(1): 27-36. http://dx.doi.org/10.1016/S0006-3207(02)00129-5

MY COTAXON

July-September 2016— Volume 131, pp. 527-533

http://dx.doi.org/10.5248/131527

Lithothelium bermudense sp. nov.,

a new Saxicolous lichen from Bermuda

FRANZ BERGER! , SCOTT LAGRECA” & ANDRE APTROOT?

' Raiffeisenweg 130, A-4794, Kopfing, Austria

? Plant Pathology Herbarium, School of Integrative Plant Science,

334 Plant Science Building, Cornell University, Ithaca, New York 14853-5904 USA

°ABL Herbarium, G.v.d.Veenstr. 107, NL-3762 XK Soest, The Netherlands

* CORRESPONDENCE TO: flechten. berger@aon.at

AsstTRAcT—Lithothelium bermudense is described as a new saxicolous lichen from Bermuda,

characterized mainly by its endolithic thallus and red-brown 3-distoseptate spores with

diamond-shaped lumina.

Key worps—endemic, Pyrenulaceae, taxonomy, Walsingham

Introduction

The genus Lithothelium Mill. Arg. comprises at least 33 species of

pyrenocarpous lichens in the Pyrenulaceae (Aptroot 2006, 2007, McCarthy 2015,

Gueidan et al. 2016). It is characterized by distoseptate ascospores with rounded

lumina, unbranched paraphyses (reported to be anastomosing in one species)

and cylindrical asci that have a conspicuous ocular chamber. The ocular chamber

is often rounded, but in some species (or specimens) it is decidedly invaginated

(sagittiform). It cannot, however, be ruled out that this invagination is simply a

more mature developmental stage, as it seems to be more common in asci with

mature spores, just prior to ascospore release. Most species occur in the tropics;

only a few are widely distributed or locally abundant. As the name already suggests

(the prefix “Litho-“ derived from the Greek word lithos, meaning “stone”), some

species occur on rock, a characteristic otherwise unknown in this family.

As part of an ongoing revision of Bermudian lichens by the first two authors

(Berger & LaGreca 2014), a lichen was collected in 2007 that was recognized by

528 ... Berger, LaGreca & Aptroot

the third author as a new species of Lithothelium. The islands that make up the

Bermuda archipelago—basically fossilized dunes (Vacher & Rowe 1997)—are

roughly 900,000 years old and among the most isolated islands in the world,

leading to the evolution of a number of endemic organisms (Thomas 2004).

The new Lithothelium species described here was collected in the Walsingham

Nature Reserve (Hamilton Parish), one of Bermudas last remaining primary-

growth forests. It is one of the last places on Bermuda where one can find

large specimens of the endemic Bermudian trees, Cassine laneana (Bermuda

olivewood) and Sabal bermudana (Bermuda sabal), which were (together with

Juniperus bermudiana, the Bermuda cedar) at one time the dominant trees

in this wet evergreen jungle. The area has escaped the strong developmental

pressures that have altered so much of the archipelago by virtue of its rough

terrain of hard limestone, which has broken through the soil surface in many

places due to the partial collapse of the ceilings of caves below. As this area also

lies in the lee of the prevailing westerly winds, it is somewhat protected from

storms, which contributes to its status as the most undisturbed, pristine forest

on Bermuda. Walsingham is home to Bermuda’ most diverse and important

cryptogam communities (Britton 1918, Rendle 1936), including some endemic

species of ferns and lichens. Indeed, the new species described below appears to

be endemic to Bermuda and grows in close association with other characteristic

Bermudian lichens, as elaborated below.

Materials & methods

This paper is based on specimens collected in 2007 by the first two authors,

deposited in the Herbarium, Department of Botany, The Natural History Museum,

London, U.K. (BM), the Herbarium, Adviesbureau voor Bryologie en Lichenologie,

Soest, the Netherlands (ABL), and the private herbarium of F. Berger (hb Berger).

The specimens were examined with an Olympus SZX7 stereomicroscope and an

Olympus BX50 compound microscope with interference contract, connected to a

Nikon Coolpix digital camera. Microscope sections were cut by hand and mounted in

water. Spores were also examined in 10% KOH.

Taxonomy

Lithothelium bermudense F. Berger, LaGreca & Aptroot, sp. nov. FIGURES 1-3

MycoBank MB 815588

Differs from Lithothelium australe by its clavate asci and its diamond-shaped spore

lumen; from L. austropacificum by its calcareous substrate, its lack of a visible thallus,

and its euseptate ascospores; and from L. echinatum by possessing larger spores that lack

spiky ornamentation.

Type: Bermuda, Hamilton Parish, Walsingham Nature Reserve, Tom Moore Trail,

32°20.7’N 64°42.75’W, alt. 3 m, on hard limestone, 2 November 2007, F. Berger 22300 &

S. LaGreca (Holotype, BM000921379; isotypes, ABL, hb Berger).

Lithothelium bermudense sp. nov. (Bermuda) ... 529

Fig. 1. Lithothelium bermudense (holotype, BM000921379).

Scale bar = 0.5 mm.

EryMoLocy: named for the Bermuda Isles, the UK Overseas Territory where the first

specimens were collected.

DESCRIPTION — THALLUS endolithic, appearing as a pink to pale brownish spot

on calcareous rock, up to 2 cm diam., prothallus absent; algae Trentepohlia,

cells c. 8 um diam. Ascomata solitary, black, closed, conical, %-74-immersed,

leaving black pits, smooth, 0.4-0.6 mm diam; walls brittle like charcoal;

excipulum and involucrellum fully integrated, the latter throughout carbonized,

combined wall c. 150-200 um thick above and at the sides, c. 100 um thick

below the asci, KOH-. OstroLum central, apical, inconspicuous, hymenium

not inspersed with oil globules, gel IKI-. Parapuyszs unbranched, slender, c.

1-1.5 um wide, no periphyses. Asci subcylindrical to clavate, 110 x 25-30 um,

fissitunicate, with a rounded ocular chamber (at least when the ascospores are

mature), wall to 5 um thick, containing 6-8 ascospores. AScosPorEs biseriate,

fusiform with protruding brighter tips, reddish-brown, darkening in KOH,

initially with 1 euseptum, later with 2 additional, incomplete eusepta and

always with 3 distosepta, endospore c. 2 um thick, terminal lumina distinctly

smaller, and protruding into the apical exospore, inner lumina diamond-

shaped, 20-25 x 7-8 um, with an open tubulus between the outer and inner

530 ... Berger, LaGreca & Aptroot

Fig. 2. Lithothelium bermudense (holotype, BM000921379).

A. Clavate ascus with young ascospores. B. Ascus with mature ascospores. Scale bars = 10 um.

lumina which is present until maturity, gelatine coat (perispore) absent, spore

wall not ornamented, spores becoming dark brown and finally shrinking (when

over-mature). PYCNIDIA not observed.

EcoLoGy & DISTRIBUTION — Grows together with Jonaspis tropica Riddle,

Stromatella bermudana (Riddle) Henssen, and small liverworts on low, sheltered,

limestone outcrops in primary coastal forest. Associated species include Strigula

bermudana (Tuck. ex Nyl.) R.C. Harris, Gyalecta farlowii Tuck., Toninia sp.,

Lithothelium bermudense sp. nov. (Bermuda) ... 531

Fig. 3. Lithothelium bermudense (isotype, hb Berger).

Ascospores, showing increasing maturity from left to right. Scale bar = 10 um.

Opegrapha sp., Cryptothecia striata G. Thor, Lempholemma lingulatum (Tuck.)

Henssen, Collema thamnodes Tuck. ex Riddle, Leptogium austroamericanum

(Malme) C.W. Dodge, and Paulia sp. Known from one specimen from the

Walsingham Nature Reserve, Bermuda.

Notes — This new species superficially resembles endolithic Verrucaria spp.,

but is easily distinguished by the red-brown, 3-distoseptate spores. According

to Aptroot (2006), it would key out as Lithothelium australe Aptroot &

H. Mayrhofer from Chatham Island, New Zealand, but the new species differs

from L. australe by possessing a different type of spore lumen, as well as clavate

asci. Lithothelium bermudense is most similar to another Lithothelium species

with brown ascospores with diamond-shaped lumina, L. austropacificum P.M.

McCarthy, from Lord Howe Island, Australia. However, L. austropacificum

lacks eusepta, occurs on basalt, and possesses a conspicuous thallus (McCarthy

1996). One more diagnostic feature of L. bermudense is the protruding

terminal lumen, the result of a spore ontogeny that is unique among saxicolous

Lithothelium species. Young spores always have three distosepta perforated at

each side by a tubulus between the inner and terminal lumina; at maturity, by

comparison, one euseptum is always present in the middle of the spores, but

never touches the external walls (Fics 2B, 3). Overmature spores inevitably lose

the tubulus, and two more discrete eusepta between the lumina are developed.

In addition, the spore colour darkens with ongoing maturity.

There is another endolithic lichen more often encountered on calcareous

substrates in Bermuda, Strigula bermudana (Berger & LaGreca 2014), which

is very similar in appearance to L. bermudense. Microscopical investigation,

however easily reveals the differences: the spores of S. bermudana are always

hyaline, euseptate, and incised at the septum, 14-16 x 4-5.9 um; ie., the spores

are much smaller, and narrower in shape, compared with L. bermudense.

532 ... Berger, LaGreca & Aptroot

Discussion

Lithothelium species differ mainly in ascospore size and pigmentation (from

hyaline to red brown), relative sizes and forms of the lumina, corticolous vs.

saxicolous habit, immersed vs. superficial thallus, and apical vs. lateral ostiole

position. The ascomata can be solitary or clustered in groups and with or

without a common ostiole. Species with brown ascospores resemble members

of the genus Pyrenula Ach., which differ by having brown to grey ascospore

pigmentation (not red brown) and lumina that are angular (not rounded)

when the ascospores are mature. The fact that mature Lithothelium ascospores

resemble immature ascospores of some Pyrenula species suggests that neoteny

is an evolutionary theme in the Pyrenulaceae (Aptroot 2006). In its current

circumscription, Lithothelium is polyphyletic (Gueidan et al. 2016).

According to previous studies of Lithothelium (Aptroot 2006, 2007), as well

as the worldwide key provided by Aptroot (2006: 544), only two previously

described species possess the combination of coloured ascospores, an

endolithic thallus, and occurrence on calcareous rocks. The more similar to

L. bermudense is the Australasian L. australe (Aptroot & H. Mayrhofer 1991);

while the dimensions of all anatomical features of these two species overlap,

the difference in spore ontogeny (as reported above) is clear. The other similar

endolithic species found on calcareous rock, L. echinatum Aptroot, occurs in

China; it differs in having smaller spores with a spiky ornamentation.

Most species of Lithothelium are uncommon, and relatively inconspicuous.

They often possess immersed, whitish thalli and so are frequently mistaken

for non-lichenized fungi in the field. Because of this, they are undercollected

by lichenologists, except in some parts of North America and tropical Asia,

where some species are locally abundant. Because of the scattered collections

of so many species, and the fact that most species are still known only from

one country (or even one specimen), it is impossible to estimate the level of

endemism in various geographic regions, or the centre(s) of diversity of the

whole genus. As limestone outcrops are relatively scarce in the tropics, however,

tropical species such as L. bermudense—assuming it only occurs on calcareous

rock—may indeed have a very restricted geographic distribution.

Acknowledgments

This work was supported by a grant to FB and SLG from the Bermuda Zoological

Society. Christine Watlington is warmly thanked for generously providing

accommodation for SLG, and for assisting with all aspects of our fieldwork. Jack Ward

graciously provided access to, and fieldwork assistance in, Walsingham Nature Reserve.

Adam Flakus (KRAM) and Roger Rosentreter (SRP) provided helpful reviews of the

manuscript prior to publication. Erich Zimmermann is kindly thanked for providing

Lithothelium bermudense sp. nov. (Bermuda) ... 533

the photos. This is contribution number 143 of the Bermuda Biodiversity Project (BBP),

Bermuda Aquarium, Natural History Museum and Zoo.

Literature cited

Aptroot A. 2006. Three new species of Lithothelium (Pyrenulaceae) from China and Thailand, with

a revised world key and annotated list of species. Lichenologist 38: 541-548.

Aptroot A. 2007. New species, combinations, lectotypifications and synonyms on Australian

Pyrenulaceae. Australasian Lichenology 60: 34-41.

Aptroot A, Mayrhofer H. 1991. Lithothelium australe spec. nov., a new lichen from New Zealand.

Mycotaxon 41: 219-222.

Berger FE, LaGreca S. 2014. Contributions to the lichen flora of Bermuda—part I. New records,

new combinations, and interesting collections of lichenized ascomycetes. Evansia 31: 41-68.

http://dx.doi.org/10.1639/079.031.0203

Britton NL. 1918. The flora of Bermuda. New York: Scribner's.

Gueidan C, Aptroot A, Caceres ME, Nguyen QB. 2016. Molecular phylogeny of the tropical lichen

family Pyrenulaceae: contribution from dried herbarium specimens and FTA card samples.

Mycological Progress 15: 7. http://dx.doi.org/10.1007/s11557-015-1154-8

Rendle AB. 1936. Notes on the flora of the Bermudas. Journal of Botany 74: 42-50.

McCarthy PM. 1996. Lithothelium austropacificum sp. nov. (Pyrenulaceae) from Lord Howe Island,

Australia. Lichenologist 28: 290-294.

McCarthy PM. 2015. A new species of Lithothelium (lichenized Ascomycota, Pyrenulaceae) from

the Tarkine region, north-western Tasmania. Telopea 18: 167-170.

http://dx.doi.org/10.7751/telopea8890

Thomas MLH. 2004. The natural history of Bermuda. Bermuda Aquarium, Natural History

Museum & Zoo, and Bermuda Zoological Society.

Vacher HL, Rowe MP. 1997. Geology and hydrology of Bermuda. Developments in Sedimentology

54: 35-90. http://dx.doi.org/10.1016/S0070-4571(04)80022-0

MY COTAXON

July-September 2016— Volume 131, pp. 535-539

http://dx.doi.org/10.5248/131.535

Harmoniella campanaensis sp. nov. from central Chile

HuGco Maprip!”’, DANIELA TORRES! & VICTOR SILVA?

‘Center for Genomics and Bioinformatics, Mayor University &

? Science Faculty, Mayor University,

Camino La Pirdmide 5750, Huechuraba, Santiago, Chile

* CORRESPONDENCE TO: hugo.madrid@umayor.cl

ABSTRACT—A new hyphomycete, Harmoniella campanaensis, is described from submerged

dicotyledonous leaves collected in La Campana National Park, central Chile. This fungus

is the second taxon described in Harmoniella and the only one known from the Americas.

Harmoniella campanaensis differs from the generic type, H. chrysocephala, by producing

much longer conidiophores (with stipes up to 850 um) and shorter conidia (3-5 um long).

KEY worps—conidial fungi, sympodial development, taxonomy

Introduction

During June 2015, fieldwork was carried out in central Chile with the

aim to collect saprophytic microfungi from soil and plant material. One of

the sampled sites was La Campana National Park (Valparaiso Region), a

vast protected area with a Mediterranean climate and a rich endemic flora

consisting of a hygrophilous laurifolious forest dominated by species such as

Aristotelia chilensis (Molina) Stuntz, Beilschmiedia miersii (Gay) Kosterm.,

and Cryptocarya alba (Molina) Looser (Elértegui & Moreira-Mufioz 2002).

Although we were mainly interested in graminicolous helminthosporioid

genera (Madrid et al. 2014, Manamgoda et al. 2014), many other interesting

hyphomycetes were also found, including new species and new records.

One interesting fungus was obtained from submerged dicotyledonous leaves.

Under the stereomicroscope, it showed conspicuous erect conidiophores with a

vesicle at the apex and produced masses of dry ochraceous conidia resembling

an Aspergillus species. However, the fungus produced conidiophores with

536 ... Madrid, Torres & Silva

strongly pigmented stipes and a tendency to proliferate percurrently through

old vesicles. A more detailed observation with light microscopy showed

that it produced septate conidiophores and sympodial, strongly denticulate

conidiogenous cells. Morphological features place this taxon in the monotypic

genus Harmoniella V.N. Boriss. (Borisova 1981, Seifert et al. 2011). However,

the dimensions of conidiophores and conidia and the substrate from which

it was isolated clearly indicate that it is different from the generic type,

H. chrysocephala V.N. Boriss.

Materials & methods

Submerged decaying leaves, twigs, and wood were collected in a stream in La

Campana National Park, Valparaiso Region, Central Chile. Samples were placed

in sterilized polyethylene bags, transported to the laboratory and kept at 4°C until

processed. The samples were placed in moist chambers, incubated at room temperature

and observed weekly for two months. Isolation of the studied Harmoniella species was

attempted by transferring conidia from conidiophores on the natural substrate to Petri

dishes containing either cornmeal agar, oatmeal agar, Sabouraud agar (all manufactured

by Becton Dickinson, USA), or water agar; the water agar plates were prepared with or

without sterilized pine needles or maize leaves. The morphological study was carried

out from lactophenol mounts using a Zeiss AxioScope A1 light microscope. A specimen

was deposited in the Herbarium of the National Natural History Museum, Santiago,

Chile (SGO).

Taxonomy

Harmoniella campanaensis Madrid, D. Torres & V. Silva, sp. nov. PLATE 1

MycoBAnk MB 815759

Differs from Harmoniella chrysocephala by its much longer conidiophores, which often

proliferate percurrently, and by its shorter conidia.

Type: Chile, Marga Marga Province, La Campana National Park, El Granizo area, on

submerged decaying dicotyledonous leaves, 30 Jun 2015, H. Madrid (Holotype, SGO

166366, dried leaf).

ETYMOLOGY: campanaensis refers to the place where the fungus was collected, La

Campana National Park.

VEGETATIVE HYPHAE septate, branched, light olivaceous brown, smooth,

up 4 um wide. CONIDIOPHORES macronematous, mononematous, solitary

to caespitose, often proliferating percurrently, straight to slightly flexuous,

septate, light olivaceous brown to dark reddish brown, thick-walled, with a

PLATE 1. Harmoniella campanaensis (holotype, SGO 166366). A. Sampling area in La Campana

National Park; B. Conidiophores on the natural substrate; C. Conidiophore showing percurrent

proliferation (arrow); D. Vesicle and conidiogenous cells; E. Sympodial conidiogenous cells with

denticles (arrow); F. Conidia. Scale bars: C = 50 um; D = 10 um; E, F = 5 um.

307,

Harmoniella campanaensis sp. nov. (Chile) ...

538 ... Madrid, Torres & Silva

subcylindrical stipe and a well-developed vesicle at the apex; stipe 688-850

x 11.5-22 um, sometimes broadened up to 35 um toward the base; vesicle

globose to depressed-globose, light olivaceous brown when young, dark brown

at maturity, 44-59.5 um wide. CONIDIOGENOUS CELLS covering the vesicle

radially, forming a compact single layer, subcylindrical, 12-18.5 x 3-4 um,

with numerous denticles up to 2 um long. Conrp1a dry, aseptate, ellipsoid to

dacryoid, verruculose to echinulate, light olivaceous to light olivaceous brown,

ochraceous in mass, 3-5 x 2.5-4 um.

ComMENTs: All attempts to obtain this fungus in culture were unsuccessful,

despite the use of different media. Therefore, no DNA sequence analyses could

be carried out and its phylogenetic placement remains unknown. Borisova

(1981), in the description of the type species, also pointed out its inability to

grow in vitro. Harmoniella chrysocephala was described from pine needles in

Yalta, Ukraine, and we could not find any record of this species from a different

country; the Global Biodiversity Information Facility (www.gbif.org) yields no

records of H. chrysocephala outside the type locality. Harmoniella chrysocephala

differs from H. campanaensis by its much smaller conidiophores (150-170 x

12-17 um) and longer conidia (6-7.2 x 3.6-4.8 um), which are more irregular

in shape (Borisova 1981). In addition to these differences, the dissimilar habitats

(pine needles versus submerged dicotyledonous leaves) and the tendency of the

Chilean fungus to develop percurrently proliferating conidiophores prompted

us to propose this organism as a taxonomic novelty.

Harmoniella has been listed, with some doubt, as a possible synonym of

Pseudogibellula Samson & H.C. Evans (Seifert et al. 2011). Pseudogibellula is

monotypic and based on Gibellula formicarum Mains [= P. formicarum (Mains)

Samson & H.C. Evans], the asexual state of Torrubiella pseudogibellulae Samson

et al., a member of Clavicipitaceae (Mains 1949, Samson & Evans 1973). We

prefer to treat Harmoniella as a separate genus due to important ecological

and morphological differences. Pseudogibellula is an entomopathogen that

attacks insects and arachnids whereas Harmoniella species are associated

with decaying plant material (Seifert et al. 2011). Furthermore, the structure

of the conidial apparatuses is quite different; Harmoniella has mononematous

smooth conidiophores with wide vesicles, which are covered directly by

conidiogenous cells, while Pseudogibellula produces synnemata and verrucose

conidiophores with very small vesicles (4-6 um wide), covered by two series of

metulae (Samson et al. 1988). Conidiogenous cells are also dissimilar, showing

conspicuous denticles in Harmoniella but only minute scars in Pseudogibellula

(Seifert et al. 2011).

Harmoniella campanaensis sp. nov. (Chile) ... 539

Acknowledgments

We would like to thank Drs. David Minter (CABI, Wallingford, England) and Keith

Seifert (Agriculture and Agri-Food Canada, Ottawa, Canada), and the Cyberliber

website (http://www.cybertruffle.org.uk/cyberliber/) for providing information and

literature about Harmoniella and related taxa. We would also like to thank Dr. José Luis

Rodriguez (Clinica Las Condes, Santiago, Chile) for translating taxonomic literature in

Russian to Spanish, and Drs. Josepa Gené and Josep Cano (Universitat Rovira i Virgili,

Reus, Spain) for critically reviewing this work and providing helpful suggestions about

it. Corporacién Nacional Forestal (CONAF), Chile, is acknowledged for providing

permission to collect biological samples in La Campana National Park. Hugo Madrid was

funded by Comision Nacional de Investigacién Cientifica y Tecnoldgica (CONICYT),

Fondo Nacional de Desarrollo Cientifico y Tecnol6gico (FONDECYT), Chile, project

no. 11140562.

Literature cited

Borisova VN. 1981. Harmoniella, a new genus of hyphomycetes. Mikol. Fitopat. 15: 89-92.

Eldortegui S, Moreira-Mufoz A. 2002. La Campana National Park: origin of a Biosphere Reserve in

central Chile. Taller La Era, Santiago.

Madrid H, da Cunha KC, Gené J, Dijksterhuis J, Cano J, Sutton DA, Guarro J, Crous

PW. 2014. Novel Curvularia species from clinical specimens. Persoonia 33: 48-60.

http://dx.doi.org/10.3767/003158514X683538

Mains EB. 1949. New species of Torrubiella, Hirsutella and Gibellula. Mycologia 41: 303-310.

http://dx.doi.org/10.2307/3755112

Manamgoda DS, Rossman AY, Castlebury LA, Crous PW, Madrid H, Chukeatirote E, Hyde KD.

2014. The genus Bipolaris. Stud. Mycol. 79: 221-288.

http://dx.doi.org/10.1016/j.simyco.2014.10.002

Samson RA, Evans HC. 1973. Notes on entomogenous fungi from Ghana. I. The genera Gibellula

and Pseudogibellula. Acta Bot. Neerl. 22: 522-528.

http://dx.doi.org/10.1111/j.1438-8677.1973.tb00873.x

Samson RA, van Reenen-Hoekstra ES, Evans HC. 1988. New species of Torrubiella (Ascomycotina:

Clavicipitales) on insects from Ghana. Stud. Mycol. 31: 123-132.

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

CBS Biodiversity Series 9. 997 p.

MY COTAXON

July-September 2016—Volume 131, pp. 541-545

http://dx.doi.org/10.5248/131.541

Rosellinia hainanensis sp. nov.

and three Rosellinia species new to China

WEI LI & LIN Guo*

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

“ CORRESPONDENCE TO: guol@im.ac.cn

ABSTRACT—A new species Rosellinia hainanensis is proposed, and three other species,

R. compacta, R. eucalypticola, and R. siggersii, are described as new to China. The four species

were collected from Hainan and Yunnan provinces.

KEY woRDS—Ascomycota, pyrenomycetous fungi, Xylariaceae, taxonomy

Introduction

Thirty-six species of Rosellinia have previously been reported from China,

but 15 of them (including R. mycophila, R. pulveracea, and R. sordaria) have

been transferred to other genera such as Amphisphaerella and Coniochaeta.

Consequently, only 21 species currently accepted in Rosellinia have been

recorded in China (Teng 1963; Tai 1979; Ju & Rogers 1990, 1999; Yuan & Zhao

1993; Lu et al. 2000; Liu et al. 2010; Petrini 2013a,b; Li & Guo 2015). Here we

describe a new species collected in Hainan and report three other species new

to China.

Materials & methods

The specimens were examined macroscopically with the aid of a Motic SMZ-168

dissecting microscope. Microscopic features of asci and ascospores were described

from slide preparations mounted in water and Melzer’s reagent, and measurements

were made from 20 ascospores, using a Zeiss Axioskop 2 plus compound microscope.

Photographs of stromata were taken using a Sony NEX-7. All specimens have been

deposited in the Herbarium Mycologicum Academiae Sinicae, Beijing, China

(HMAS).

542 ... Li & Guo

Taxonomy

Rosellinia hainanensis Wei Li bis & L. Guo, sp. nov. Fics 1-3

FUNGAL NAME EN 570257

Differs from Rosellinia spiraliterfissurata by its shorter and broader ascospores and from

R. chusqueae by its higher ascus apical ring.

Type: China, Hainan, Changjiang, Bawangling, alt. 603 m, on little twigs of unidentified

plant, 11.XII.2009, Y. F Zhu & L. Guo 123 (Holotype, HMAS 269892).

ErymMo ocy: The epithet refers to the type locality.

Stromata solitary or crowed, pear shaped, surface coarse and black, 0.5-0.74

mm high, 0.5-0.9 mm wide. Ostioles irregular, indistinctly papillate, black.

Subiculum black, reduced to stroma base, evanescent. Ectostroma 60-80

um thick, black. Entostroma disappearing at maturity. Asci eight-spored,

cylindrical, the spore bearing part 125-145 um long, 12-15 um broad, with

apical ring 7-10 um high, 3-5 um broad and bluing in Melzer’s iodine reagent.

Ascospores dark brown, surrounded by a slimy sheath, asymmetrically

ellipsoidal, with broadly rounded ends, 21-24 x 11-13 um, with sigmoid germ

slit slightly shorter than spore-length.

ComMENTSs: Rosellinia hainanensis is similar to Rosellinia spiraliterfissurata

L.E. Petrini, which differs by having longer and narrower ascospores (26.5-34

x 7-11 um; Petrini 2013a,b). Rosellinia chusqueae Pat. differs from Rosellinia

hainanensis by its shorter apical ring (6.7-7 um high; Petrini 2013b).

Rosellinia hainanensis is described in Rosellinia (rather than Helicogermslita)

because the holotype lacks the helical ascospore germ slit that is the most

distinctive character of Helicogermslita.

Rosellinia compacta Takemoto, Mycologia 101: 89, 2009. FIGS 4-5

Stromata crowed, subglobose, copper brown, black, 0.8-1.2 mm _ high,

0.9-1.2 mm wide. Ostioles papillate, black. Subiculum dark brown. Ectostroma

80-140 um thick, black. Entostroma brown. Perithecia subglobose, 550-660

um high, 700-920 um wide. Asci eight-spored, cylindrical, 252-275 um total

length, 8-13 um broad, the spore bearing part 220-229 um long, with apical

ring bluing in Melzer’s iodine reagent, with rounded or angular bulge at

upper rim, 9-12 um high, 3.5-7 um broad. Ascospores light brown or brown,

surrounded by a slimy sheath, asymmetrically fusiform-shaped, with thread-

Fics 1-9. Rosellinia hainanensis (HMAS 269892, holotype). 1. Stromata on twig; 2. Ascus and

ascospores; 3. Ascospore. Rosellinia compacta (HMAS 253036). 4. Stromata on wood; 5. Ascus

and ascospores. Rosellinia eucalypticola (HMAS 245220). 6. Stromata on wood; 7. Ascus and

ascospores. Rosellinia siggersii (HMAS 253035). 8. Stromata on wood; 9. Ascospore.

Rosellinia hainanensis sp. nov. [China] ... 543

544 ... Li& Guo

like ends, 41-49(-55) x (5-) 6-9 um, with a longitudinal straight germ slit

spore-length in the center.

SPECIMENS EXAMINED: China, Yunnan, Mengla, Menglun, Xishuangbanna Tropical

Botanical Garden, Chinese Academy of Sciences, alt. 570 m, on corticated wood,

16.X.2013, W. Li 2930 (HMAS 253039); W. Li 2949 (HMAS 253036).

ComMENTSs: Rosellinia arcuata Petch differs from R. compacta by larger stromata

(0.9-2.75 mm high, 1.1-2.5 mm wide; Petrini 2013b); and Rosellinia necatrix

Berl. ex Prill. differs from R. compacta by smaller ascospores (30.6-46.8 x

5.4-8.1 uum) and larger stromata (1-1.9 mm high, 1.2-2 mm wide; Petrini

2013b).

Rosellinia eucalypticola Henn. & E. Nyman, Monsunia 1: 166, 1899. FIGs 6-7

Stromata solitary or 2-4 gregarious, globose or subglobose, surface smooth

and black, 0.44-0.7 mm high, 0.66-1 mm wide. Ostioles papillate, black.

Subiculum reduced to stroma base, evanescent. Ectostroma 40-80 um thick,

black. Entostroma brown. Perithecia subglobose, 400-500 um high, 560-700

um wide. Asci eight-spored, cylindrical, 100-120 um total length, 6-7.5 um

broad, the spore bearing part 60-67 um long, with apical ring bluing in Melzer’s

iodine reagent, 1.5-2 um high, 1.5-3 um broad. Ascospores light brown or

brown, asymmetrically ellipsoidal, with broadly rounded ends, 8.5-11 x

4.5-5.5 um, with straight germ slit as long as or slightly shorter than spore.

SPECIMENS EXAMINED: China, Hainan, Changjiang, Bawangling, alt. 650 m, on

decorticated wood, 10.1.2009, W. Li, X.Y. Liu 2216 (HMAS 245220).

Rosellinia siggersii L.E. Petrini, Index Fungorum 25: 4, 2013. Fics 8-9

Stromata crowded, subglobose, semiglobose to cupulate, sometimes with a

broad, short stipe, dark brown, 1.4-1.8 mm high, 1.4-1.6(-2) mm wide. Ostioles

papillate to conical, black. Subiculum woolly, dark red brown, with synnemata.

Ectostroma 80-160 um thick, black. Entostroma white or cream, disappeared

at maturity. Perithecia subglobose, 900-1000 um high, 1300-1500 um wide.

Asci eight-spored, cylindrical, 277-365 um total length, 9-10 um broad, the

spore bearing part 200-252 um long, with apical ring bluing in Melzer’s iodine

reagent, ampulliform, 12-12.5 um high, 4-7 um broad. Ascospores light brown

or brown, asymmetrically fusiform-shaped, with thread-like ends, 41-46(-52)

x 5-8 um, with straight germ slit on flat side, 9-13 um long, shorter than spore.

SPECIMENS EXAMINED: China, Yunnan, Jinghong, Dadugang, alt. 1300 m, on corticated

wood, 15.X.2013, W. Li 3104 (HMAS 253035).

ComMENTSs: Rosellinia siggersii differs from R. acutispora (Theiss.) L.E. Petrini

by its ascospores with thread like, elongated and pinched ends (Petrini 2013a,b).

Rosellinia hainanensis sp. nov. [China] ... 545

Discussion

China is a vast country with many different geographical zones, and it is

probable that continuing field expeditions and study will reveal additional

Rosellinia species.

Acknowledgments

The authors would like to express their deep thanks to Drs. L. Vasilyeva (Vladivostok,

Russia) and L.E. Petrini (Breganzona, Switzerland) for serving as pre-submission

reviewers, to Dr. Shaun Pennycook (Auckland, New Zealand) for nomenclatural review,

to the support by the Ministry of Science and Technology of the People’s Republic of

China (No. 2013FY110400).

Literature cited

Ju YM, Rogers JD. 1990. Astrocystis reconsidered. Mycologia 82(3): 342-349.

http://dx.doi.org/10.2307/3759905

Ju YM, Rogers JD. 1999. The Xylariaceae of Taiwan (excluding Anthostomella). Mycotaxon 73:

343-440.

Li W, Guo L. 2015. Rosellinia brunneola sp. nov. and R. beccariana new to China. Mycotaxon 130:

233-236. http://dx.doi.org/10.5248/130.233

Liu CE, Lu T, Gao JM, Wang MQ, Lu BS. 2010. Two new Chinese records of Rosellinia. Mycosystema

29(3): 459-462.

Lu BS, Hyde KD, Ho WH. et al. 2000. Checklist of Hong Kong Fungi. Fungal Diversity Research

Series,.5; 1-207,

Petrini LE. 2013a. Nomenclatural novelties : Liliane Petrini. Index Fungorum 25: 1-6,.

Petrini LE. 2013b. Rosellinia - a world monograph. Bibliotheca Mycologica 205. 410 p.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

Yuan ZQ, Zhao ZY. 1993. Studies on the Genera Amphisphaerella, Coniochaeta and Rosellinia of

XingJiang, China. Acta Mycologica Sinica 12(3): 180-186.

MY COTAXON

July-September 2016—Volume 131, pp. 547-551

http://dx.doi.org/10.5248/131.547

Phaeomonilia nanningensis sp. nov. and

a new Craspedodidymum record from southern China

CHUN-LING YANG, JIN-YE WANG, JI-WEN XIA,

YING-Ru1 Ma, JIAN-MEI GAO & XIU-GUO ZHANG*

Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong 271018, China

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

ABSTRACT—A new species, Phaeomonilia nanningensis, is described and illustrated from

specimens collected on dead branches in Guangxi Province. The fungus is characterized

by ellipsoid, catenate, pale brown to brown conidia formed by thallic disarticulation of

fertile branches on macronematous conidiophores. A key to the four described species of

Phaeomonilia is also provided. Craspedodidymum abigianense is newly recorded from China.

Key worps—conidial fungi, taxonomy

Introduction

Phaeomonilia was established by Castafieda-Ruiz et al. (2007) with two

species: P pleiomorpha R.F. Castaneda et al. (type species) and P corticola

(R.E Castafieda) R.E Castafieda et al. [= Monilia corticola R.F. Castaneda].

Subsequently, Ma et al. (2015) described a third species, P guangxiensis

Y.R. Ma & X.G. Zhang. The genus is mainly characterized by macronematous

conidiophores with unicellular globose colorless conidia formed by thallic

disarticulation of fertile branches and by the Stylaspergillus-like synanamorph.

It is similar to Botryomonilia Goos & Piroz. (Goos & Pirozynski 1975) in its

branching system, which narrows progressively towards the apex before the

maturation, and the fragmentation or disarticulation process that produces

the thallic—arthric conidia, as described by Kendrick (2003). Botryomonilia,

however, produces conidia that are connected by an isthmus (Castaneda-Ruiz

et al. 2007).

During our ongoing survey of fungi associated with woody debris in the

tropical forests of Guangxi and Hainan provinces, two species were collected

548 ... Yang & al.

on decaying twigs and dead stems. One of them, with morphological

characteristics of Phaeomonilia (Castafhieda-Ruiz et al. 2007), is described here

as a new species, P. nanningensis; the other was identified as Craspedodidymum

abigianense, a new record for China. Voucher specimens are conserved in the

Herbarium, Department of Plant Pathology, Shandong Agricultural University,

Taian, China (HSAUP) and Herbarium Mycologium, Chinese Academy of

Sciences, Beijing, China (HMAS).

Phaeomonilia nanningensis C.L. Yang & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 818085

Differs from Phaeomonilia corticola by its larger, ellipsoid conidia; and from

P. guangxiensis by its larger, pale brown to brown, ellipsoid conidia.

Type: China, Guangxi Province, Nanning, Longhushan, on dead stems of unidentified

broadleaf tree, 8 Dec. 2015, C.L. Yang (holotype, HSAUP H7858; isotype, HMAS

245606).

ETYMOLOGY: nanningensis, in reference to the type locality, Nanning.

Co.Lonies on the natural substratum effuse, circumvolute, twinkling, brown.

Mycelium mostly superficial but some immersed; superficial hyphae pale brown,

branched, septate, cylindrical, smooth. CONIDIOPHORES macronematous,

mononematous, erect, straight or flexuous, cylindrical, septate, smooth,

about 410 um long, 9-10.5 um diam. at the base, brown to pale brown, mostly

dichotomously branched towards the apex. CONIDIOGENOUS CELLS holothallic,

determinate, inwardly brown to pale brown, smooth, lageniform to subulate.

RAMOCONIDIA AND cConipiIA thallic-arthric, catenate, ellipsoid, broadly and

more or less Y-shaped, 13.5-25 x 9-13 um, brown to pale brown, dry, smooth,

forming by disarticulation of the conidiogenous branches.

ComMENTs - Phaeomonilia nanningensis slightly resembles P. corticola and

P. guangxiensis in conidial shape. However, compared with P. nanningensis, the

conidia of P corticola are ellipsoid to doliiform and smaller (5.5-16.0 x 4-5 um;

Castaheda-Ruiz et al. 2007); and the conidia of P. guangxiensis are colorless,

ellipsoid to globose, and smaller (11-14.5 x 7-11 um; Ma & Zhang 2015).

Key to species of Phaeomonilia

1. Stylaspergillus-like synanamorph present ...................00005 P. pleiomorpha

Le Nosy anal Of pi Present a... Aiviats, a. keto os Mahon kan eas ween neo oe srs eas 2

2, Goniidia clobose; 7-10 <= 14S Wi 9B ob aba eset nested eset P. guangxiensis

2. onda dolitorim OF slipsoid! haw «cea sacha + tay: bahay: batty pate yepectaehsckate rs 3

3. Conidia doliiform or ellipsoid, 5.5-16 x 4-5 um... eee eee ee eee P. corticola

3. Conidia ellipsoid and larger, 13.5-25 x 9-13 um ................. P. nanningensis

Phaeomonilia nanningensis sp. nov. (China) ... 549

Fic. 1. Phaeomonilia nanningensis (holotype, HSAUP H7858).

A, B. conidiophores and conidia. C, D. conidiogenous cells and conidia. E. conidia.

550 ... Yang & al.

A B C )

1@@

20um

Fic. 2. Craspedodidymum abigianense (HSAUP H7566).

A: conidiophore and conidium. B: conidiogenous cell and conidia. C: conidiophore. D: conidia.

20um

Craspedodidymum abigianense Lunghini & Onofri, Trans. Br. Mycol. Soc.

74: 208 (1980) FIG. 2

Colonies on the natural substratum effuse, dark brown. Conidiophores

macronematous, mononematous, straight or slightly flexuous, dark brown,

Phaeomonilia nanningensis sp. nov. (China) ... 551

4-6-septate, 120-170 x 5.2-7 um, tapered gradually towards the apex.

Conidiogenous cells terminal, ellipsoidal, monophialidic or rarely polyphialidic,

percurrent, 20-30 x 6.5-8 um, paler brown, with a distinct funnel-shaped

collarette, 1.5-2.0 x 5.5-7.5 um. Conidia acrogenous, solitary, spherical or

more often obovoid, smooth, dark brown, generally paler when young, aseptate,

15.5-18.5 x 5.5-16.5 um, papillate at the base.

SPECIMEN EXAMINED: CHINA, HAINAN PROVINCE, Diaoluoshan, on dead stems of

unidentified broadleaf tree, 26 May 2014, X.Y. Li (HSAUP H7566).

Comments - The genus Craspedodidymum was established by Holubova-

Jechova (1972), with C. elatum Hol.-Jech. as type species. Craspedodidymum

abigianense, described by Lunghini & Onofri (1980) from Céte d'Ivoire, West

Africa, is reported for the first time from China. Our Chinese specimen closely

matched the original description by Lunghini & Onofri (1980), but the type

material had longer conidiophores (225-250 x 4.5-5.5 um) and conidia that

were smaller (14.5-16.5 x 13.5-14.5 um) and slightly lighter in colour.

Acknowledgments

The authors express gratitude to Dr. Rafael E Castafteda-Ruiz and Dr. Eric H.C.

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

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

China (Nos. 31093440, 31230001, 31493010, 31493011) and the Ministry of Science and

Technology of the People’s Republic of China (Nos. 2006FY 120100).

Literature cited

Castaneda-Ruiz RF, Heredia-Abarca G, Arias RM, Saikawa M, Minter DW, Stadler M. 2007.

Anamorphic fungi from submerged plant material: Phaeomonilia pleiomorpha, P. corticola and

Cacumisporium pleuroconidiophorum. Mycotaxon 100: 327-336.

Goos RD, Pirozynski KA. 1975. Fungi of Barro Colorado Island: new and interesting hyphomycetes.

Canadian Journal of Botany 53: 2927-2932. http://dx.doi.org/10.1139/b75-322

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

Mykologie 26: 70-73.

Kendrick B. 2003. Analysis of morphogenesis in hyphomycetes: new characters derived from

considering some conidiophores and conidia as condensed hyphal systems. Canadian Journal

of Botany 81: 75-100. http://dx.doi.org/10.1139/b03-008

Lunghini D, Onofri S. 1980. Craspedodidymum abigianense sp. nov., a new dematiaceous

Hyphomycete from Ivory Coast forest litter. Transactions of the British Mycological Society

74: 208-211. http://dx.doi.org/10.1016/S0007-1536(80)80033-7

Ma YR, Xia JW, Zhang XG, Castafieda-Ruiz RF. 2015. New species of Phaeomonilia and Mirandina

from dead branches in China. Mycotaxon 130: 775-781. http://dx.doi.org/10.5248/130.775

MY COTAXON

July-September 2016— Volume 131, pp. 553-558

http://dx.doi.org/10.5248/131.553

Pseudoacrodictys from southern China:

P. ambigua sp. nov. and a new record

JIAN- MEI GAO, JIN- YE WANG, CHUN-LING YANG,

JI-WEN X14, YING-RuI Ma & XIU-GUO ZHANG

Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong 271018, China

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

ABSTRACT—Two species of Pseudoacrodictys were discovered from tropical forests in

southern China. One new species, P. ambigua, is described and compared with closely related

taxa, and P. magnicornuata is newly recorded from China.

Key worps—hyphomycetes, taxonomy

Introduction

The genus Pseudoacrodictys, typified by Acrodictys eickeri |= P. eickeri\,

was erected by Baker & Morgan-Jones (2003) to accommodate seven species

of Acrodictys. Subsequently, seven additional Pseudoacrodictys species have

been described (Somrithipol & Jones 2003, Castaheda-Ruiz et al. 2010, Zhang

et al. 2011, Rashmi & Pandey 2012, Fiuza et al. 2014, Ma et al. 2016). The

genus is characterized by single, straight or slightly flexuous, thick-walled,

cylindrical, aseptate or septate, brown to dark blackish-brown conidiophores

with an integrated, terminal, monoblastic, cylindrical, determinate or (more

commonly) indeterminate with percurrent elongate conidiogenous cells

and schizolytic conidial secession. Conidia, which are holoblastic, solitary,

acrogenous, subglobose, broadly pyriform to turbinate or somewhat irregularly

shaped, dictyoseptate with numerous obliquely arranged septa, and brown

to dark brown, commonly possess pale brown, aseptate or septate, straight,

undulate, involute to uncinate appendages.

During our ongoing survey of microfungi associated with plant debris

in tropical and subtropical forests of southern China, two species with the

554 ... Gao & al.

TABLE 1. Morphological comparison of Pseudoacrodictys species with appendaged

conidia

SPECIES

P. ambigua

P. appendiculata

P. balansae

P. corniculata

P. eickeri

P fici

P. limoniaceae

P. magnicornuata

P. viridescens

——— CONIDIA

Obclavate or irregular,

sometimes lobed

Pyriform to

turbinate

Subnapiform, turbinate

to irreg. subglobose,

sometimes lobed

Globose,

subglobose, or

broadly pyriform

Pyriform to

turbinate

Subglobose,

sometimes lobed

Turbinate to

obpyriform

Turbinate, subnapiform,

subglobose, or irregular

Globose, subglobose, or

broadly pyriform

SIZE

(um)

70-95 x

25-90

27-45 x

22-30

42-68 x

28-62

19-35 x

7-30

28-64 x

22-42

35-52 x

22-44

21-34 x

3-33

50-87 x

50-67

62-80 x

40-60

NUMBER

2-5

3-4

2-4

1-8

4-6

APPENDAGES

LENGTH

(um)

40-130

10-30

8-20

30-70

18-50

8-20

SHAPE

Subulate

Curved,

undulate,

swollen

Subulate,

curved

Cylindrical

Curved,

horn-like

Circinate

Cylindrical

morphological characteristics of Pseudoacrodictys were collected on decaying

twigs and dead stems: P ambigua is proposed here as new to science, and

P. magnicornuata represents a new record for China. Voucher specimens

are conserved in the Herbarium, Department of Plant Pathology, Shandong

Agricultural University, Taian, China (HSAUP) and Herbarium Mycologium,

Chinese Academy of Sciences, Beijing, China (HMAS).

Pseudoacrodictys ambigua J.M. Gao & X.G. Zhang, sp. nov.

MycoBank MB 8

18098

FIG. 1

Differs from all other Pseudoacrodictys species with appendaged conidia by its larger,

obclavate or irregular, and sometimes lobed conidia and its longer conidial appendages.

Type: China. Yunnan Province: tropical forest of Menglun, on dead stems of

unidentified broadleaf tree, 20 Dec. 2014, J.M. Gao (holotype, HSAUP H9624; isotype,

HMAS 245608).

EryMo.Locy: ambigua, referring to the numerous obscure oblique conidial septa.

Pseudoacrodictys ambigua sp. nov. (China) ... 555

Fic. 1. Pseudoacrodictys ambigua (holotype, HSAUP H9624).

A. conidiophores, conidiogenous cells and conidia. B. conidia.

556 ... Gao & al.

COLONIES on natural substrate effuse, dark brown. Mycelium superficial and

immersed, composed of branched, septate, brown hyphae. CONIDIOPHORES

macronematous, mononematous, erect, straight or flexuous, septate, brown to

dark brown, 45-75 x 3.5-7 um. CONIDIOGENOUS CELLS integrated, terminal,

cylindrical, monoblastic, indeterminate, with 1-2 percurrent elongations,

smooth, dark brown, truncate at the apex after schizolytic conidial secession.

ConipIA_ acrogenous, holoblastic, solitary, dry, obclavate or irregular,

dictyosporous, with numerous obscure oblique septa, thick-walled, sometimes

lobed, brown, 70-95 x 25-90 um. Basal cell cylindrical, delimited by a

transverse, cambered septum, 6.0-7.0 x 3.5-5.0 um. Appendages 2-5, aseptate,

pale brown, straight or slightly flexuous, <130 um long, 0.5-1.5 um diam. at the

conidial apex.

ComMMENTS—Pseudoacrodictys species with appendaged conidia are compared

in TABLE 1; the other eight species all differ from P. ambigua by their smaller

conidia with shorter appendages.

Pseudoacrodictys magnicornuata Fiuza, Gusmao & R.F. Castafieda-Ruiz,

Mycotaxon 127: 36 (2014) FIG. 2

COLONIES on natural substrate effuse, dark brown. Mycelium superficial and

immersed, composed of branched, septate, brown hyphae. CONIDIOPHORES

macronematous, mononematous, erect, straight or slightly flexuous,

3-5-septate, brown to dark brown, 40-65 x 4-6 um, 6-7.5 um diam. at the

base. CONIDIOGENOUS CELLS integrated, terminal, cylindrical, monoblastic,

indeterminate, with 1-2 percurrent extensions, smooth, brown, truncate at

the apex. Conrp1a holoblastic, acrogenous, solitary, turbinate, subnapiform,

subglobose, or somewhat irregularly shaped, dictyoseptate and with several

obscure septa, thick-walled, brown, 40-70 x 30-55 um, seceding schizolytically,

the basal cell 2.5-3.5 um diam. and delimited by a transverse, cambered septum.

Appendages 9-20, aseptate, curly, undulate, swollen, pale brown, 8-30 x

3-6 um, scattered around the conidia.

SPECIMEN EXAMINED: CHINA, YUNNAN PROVINCE, tropical forest of Dadugang, on

dead stems of unidentified broadleaf tree, 22 Dec. 2014, J.M. Gao (HSAUP H9677).

CoMMENTS—Pseudoacrodictys magnicornuata, which was described from

Brazil, is reported for the first time from China. The Chinese specimen closely

matched the protologue description (Fiuza et al. 2014), except that the Brazilian

type specimen had more (9-35) appendages.

Acknowledgments

The authors express gratitude to Dr. Rafael F. Castaheda-Ruiz and Dr. De-Wei Li for

serving as pre-submission reviewers and for their valuable comments and suggestions.

Pseudoacrodictys ambigua sp. nov. (China) ... 557

Fic. 2. Pseudoacrodictys magnicornuata (HSAUP H9677).

A. conidiophores, conidiogenous cells and conidia. B. conidia.

C. conidiophore and conidiogenous cell.

This project was supported by the National Natural Science Foundation of China

(Nos. 31093440, 31230001, 31493010, 31493011) and the Ministry of Science and

Technology of China (Nos. 2006FY120100).

Literature cited

Baker WA, Morgan-Jones G. 2003. Notes on Hyphomycetes. XCI. Pseudoacrodictys, a novel genus

for seven taxa formerly placed in Acrodictys. Mycotaxon 85: 371-391.

Castaneda-Ruiz RF, Heredia Abarca G, Arias RM, Stadler M, Saikawa M, Minter DW. 2010.

Anaselenosporella sylvatica gen. & sp. nov. and Pseudoacrodictys aquatica sp. nov., two new

anamorphic fungi from Mexico. Mycotaxon 112: 65-74. http://dx.doi.org/10.5248/112.65

558 ... Gao & al.

Fiuza PO, Gusmao, LFP, Cruz ACR, Castafieda-Ruiz RF. 2014. Conidial fungi from semiarid

Caatinga biome of Brazil: a new species of Pseudoacrodictys. Mycotaxon 127: 33-37.

http://dx.doi.org/10.5248/127.33

Ma YR, Xia JW, Castafieda-Ruiz RF, Zhang XG. 2016. Two new species of Pseudoacrodictys

from Hainan, China. Nova Hedwigia 102: 69-75.

http://dx.doi.org/10.1127/nova_hedwigia/2015/0303

Rashmi D, AK Pandey. 2012. Pseudoacrodictys steviae —- a new generic and species record for India.

Journal of Mycology and Plant Pathology 42: 251-253.

Somrithipol S, Jones EBG. 2003. Pseudoacrodictys dimorphospora sp. nov., a new graminicolous

hyphomycete from Thailand. Sydowia 55(2): 365-371.

Zhang YD, Ma J, Wang Y, Ma LG, Castafieda Ruiz RE, Zhang XG. 2011. New species and

record of Pseudoacrodictys from southern China. Mycological Progress 10: 261-265.

http://dx.doi.org/10.1007/s1157-010-0696-z

MY COTAXON

July-September 2016— Volume 131, pp. 559-563

http://dx.doi.org/10.5248/131.559

Capsicispora mycophila gen. & sp. nov.

from southern China

JIN-YE WANG', CHUN-LING YANG’, JI-WEN XIA’,

YING-RuI Ma’, JIAN-MErI GAo’, YU-MEI Cal” & X1U-GUO ZHANG?

‘College of Animal Science and Veterinary Medicine, Shandong Agricultural University &

Sino-German Cooperative Research Centre for Zoonosis of Animal Origin Shandong Province,

Taian, Shandong, China

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

*CORRESPONDENCE TO: sdau613@163.com, caiyum@163.com

ABSTRACT—A new hyphomycete genus and species, Capsicispora mycophila, is described

and illustrated. Capsicispora is characterized by mycophilic mycelium and mononematous,

hyaline, determinate conidiophores with monoblastic, terminal conidiogenous cells and

schizolytic secession of solitary, hyaline conidia. A key to Capsicispora and morphologically

similar genera is provided.

KEY worps—conidial fungi, hyperparasite, taxonomy

Introduction

Saprobic hyphomycetes are highly diverse on plant material and numerous

new taxa have been recently discovered (Zhang et al. 2009, Ren et al. 2012,

Li et al. 2012, Ma et al. 2014, Xia et al. 2014). During our ongoing surveys of

saprobic fungi associated with woody debris in subtropical forests of Henan

province, an undescribed mycophilic fungus was collected overgrowing the

conidiophores and conidia of Ellisembia camelliae-japonicae (Wu & Zhuang

2005). Detailed studies of its morphology and development revealed that it

could not be accommodated in any previously described genus (Carmichael

et al. 1980; Subramanian 1971; Ellis 1971, 1976; Matsushima 1975, 1983, 1985,

1993, 1995; Castaneda-Ruiz & Kendrick 1990, 1991; Wu & Zhuang 2005). For

that reason, we describe a new genus for this unknown fungus.

560 ... Wang & al.

Material & methods

Samples were processed, examined, and photographed as described in Xia et al.

(2014). Adobe Photoshop 7.0 was used to process images into compound photographs;

backgrounds were modified for esthetic reasons. We were unable to culture this species

and so no molecular data are presented here. The specimens are deposited in the

Herbarium of Department of Plant Pathology, Shandong Agricultural University, Taian,

China (HSAUP) and the Mycological Herbarium, Institute of Microbiology, Chinese

Academy of Sciences, Beijing, China (HMAS).

Capsicispora J.Y. Wang, Yu M. Cai & X.G. Zhang, gen. nov.

MycoBank MB 818082

Differs from Capsicumyces by its mycophilic habit and its schizolytic conidial secession.

TYPE SPECIES: Capsicispora mycophila J.Y. Wang et al.

EryMo_ocy: Referring to the conidial shape is similar to Capsicumyces.

MyceLium mycophilic. CONIDIOPHORES macronematous, mononematous,

hyaline, unbranched, erect. CONIDIOGENOUS CELLS monoblastic, integrated,

terminal, hyaline, determinate. Conidial secession schizolytic. CoNnip1Aa

solitary, hyaline.

Capsicispora mycophila J.Y. Wang, Yu M. Cai & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 818083

Differs from Capsicumyces delicatus by its mycophilic habit, its schizolytic conidial

secession, and its predominantly navicular or fusiform conidia with an apical

mucilaginous sheath.

Type: China, Henan Province, Liankangshan, on conidiophores and conidia of Ellisembia

camelliae-japonicae (Subram.) W.P. Wu growing on dead branches of unidentified

broadleaf tree, 16 Oct. 2015, J.Y. Wang (Holotype, HSAUP H10083; isotype, HMAS

245607).

EryMoLoey: From Greek myco = fungus, and philus = loving, indicating its occurrence

on other fungi.

CoLonigs on the natural substrate effuse, hyaline, very thin, scarcely visible,

overgrowing Ellisembia camelliae-japonicae colonies (especially overgrowing

the conidia). Mycelium creeping, mycophilic, superficial, repent, apparently

aseptate or rarely septate, hyaline, smooth, 1-2 um wide. CONIDIOPHORES

mononematous, hyaline, simple, erect, aseptate, narrowly truncate at the apex,

9-16 x 2-4 um. CONIDIOGENOUS CELLS integrated, terminal, monoblastic,

hyaline. Conidial secession schizolytic. Conrp1< solitary, hyaline, 0-1-septate,

smooth, fusiform, navicular or obclavate, 20-30 um long, 4—4.6 um diam. at the

broadest part, with a short, protruding truncate hilum at the base, attenuated

towards the apex, apex often surrounded by a hyaline, globose mucilaginous

sheath.

Capsicispora mycophila gen. & sp. nov. (China) ... 561

| [ | f |

' ij \ | yy '

d Vv ’ " \\ ’ Y \

Fic. 1. Capsicispora mycophila. (ex holotype, HSAUP H10083): A. Four conidiophores (and one

developing conidium). B. Habit on substrate (Ellisembia camelliae-japonicae - dark conidiophores

and phragmoconidia): conidiophores, conidiogenous cells, and conidia. C. Conidia.

562 ... Wang & al.

COMMENTS - Capsicispora demonstrates unique features in morphology

and ontogeny. Several existing genera (e.g., Capsicumyces, Eriomycopsis,

Parachionomyces, Paratrichoconis, and Soloacrospora) have morphological

characters that resemble those of Capsicispora (Spegazzini 1910, Ciferri 1954,

Deighton & Pirozynski 1972, Gamundi et al. 1979, Thaung 1979, Castanieda-

Ruiz & Kendrick 1991, Braun 1994, Castafeda-Ruiz et al. 1997, Braun &

Melnik 1998). However, in Eriomycopsis, conidiophores are indeterminate with

sympodial proliferations and the conidia are rostrate, while in Capsicispora the

conidiogenous cells are determinate and the conidia lack a rostrum. Septate

branched geniculate conidiophores, polyblastic conidiogenous cells, and

rostrate conidia separate Parachionomyces from Capsicispora. Paratrichoconis

and Capsicumyces can be distinguished from Capsicispora by their non-

mycophilic mycelium and rhexolytic conidia secession. The whip-like setae,

non-mycophilic mycelium, pale brown conidiophores of Soloacrospora clearly

separate that genus from Capsicispora with its mycophilic mycelium, hyaline

conidiophores, and absence of setae.

Key to Capsicispora and morphologically similar genera

Beviyceliunrmycophiet hyper parasitic a. S.-1 nx tel ox mel ot eel oN ea Stok cia teehee Seeks 2

LeAViy celine NOnsNtyeO Ph Ite Rese cy ore acetone ace nce oc nyc ec mao ences ety Miia tt nl es Wet t

DAG ONT TOSUEALEG a ir Na Norn tia Ne rntcky Necmnicke Narnahe Nacwtiole Necwile gtewiay tread kate 3 te arg 3

2 Conde norrosttates sis ses ee ke Ree eR ae Rt Re eRe SAS Capsicispora

SHC OnTMOphoOres; WraTic he ek reheat pleco EA seo inlet tae Lets Parachionomyces

3.-Conidiophores unbranched. it. oa a ee sl el oe ae Eriomycopsis

4 SSCLACNPECSCNU Sheet a Mec a ame rtar sitet ardl ee ar digeh tas Seto hae TAR gs SE rs Soloacrospora

AROELAC ADSEMr. tivena © Sine Minch Siena Meee Races 2 Wadia Meshal chats Wm os ahelah st wi 5

5.Gonidial-secession’schizolytic’ << fo 5% ig «5 'sfag beng 8 a eng 2 eleay oe e Capsicumyces

SConidial secessionsthexOlytic: «ty! fg scat chee acal eps ol ep eeal epee pa e| Paratrichoconis

Acknowledgments

The authors express gratitude to Dr. Rafael E Castafteda-Ruiz and Dr. Eric H.C.

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

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

China (Nos. 31093440, 31230001, 31493010, 31493011) and the Ministry of Science and

Technology of the People’s Republic of China (Nos. 2006FY 120100).

Literature cited

Braun U. 1994. Miscellaneous notes on phytopathogenic hyphomycetes. Mycotaxon 51: 37-68.

Braun U, Melnik VA. 1998. Eriomycopsis lasiosphaeriicola sp. nov. A new hyperparasitic

hyphomycete fom the Far East of Russia. Mikologiya i Fitopatologiya 32(1): 3-6.

Carmichael JW, Kendrick WB, Conners IL, Sigler YL. 1980. Genera of hyphomycetes. University

of Alberta Press, Alberta. 386 p.

Capsicispora mycophila gen. & sp. nov. (China) ... 563

Castafeda-Ruiz RF, Kendrick WB. 1990. Conidial fungi from Cuba: I. University of Waterloo

Biology Series 32. 53 p.

Castafieda-Ruiz RF, Kendrick WB. 1991. Ninety-nine conidial fungi from Cuba and three from

Canada. University of Waterloo Biology Series 35. 132 p.

Castafieda-Ruiz RE, Guarro J, Cano J. 1997. Notes on conidial fungi. XII. New or interesting

hyphomycetes from Cuba. Mycotaxon 63: 169-181.

Ciferri R. 1954. Schedae Mycologicae XII-XXXIV. Sydowia 8: 245-270.

Deighton FC, Pirozynski KA. 1972. Microfungi V. More hyperparasitic hyphomycetes. Mycological

Papers 128. 110 p.

Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew. 608 p.

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew.

507 p.

Gamundi IJ, Arambarri AM, Bucsinszky AM. 1979. Micoflora de la hojarasca de Nothofagus

dombeyi, Il. Darwiniana 22(1-3): 189-216.

Li DW, Kendrick B, Chen JY. 2012. Two new hyphomycetes: Codinaea sinensis sp. nov. and

Parapleurotheciopsis quercicola sp. nov., and two new records from Quercus phillyraeoides leaf

litter. Mycological Progress 11: 899-905. http://dx.doi.org/10.1007/s11557-011-0805-7

Ma LG, Xia JW, Ma YR, Castafieda-Ruiz RE, Zhang XG. 2014. Two new species of

Spadicoides and Gangliostilbe from southern China. Mycological Progress 13: 547-552.

http://dx.doi.org/10.1007/s11557-013-0937-z

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Published by the author:

Kobe, Japan. 209 p.

Matsushima T. 1983. Matsushima Mycological Memoirs No.3. Published by the author, Kobe,

Japan. 89 p.

Matsushima T. 1985. Matsushima Mycological Memoirs No.4. Published by the author, Kobe,

Japan. 68 p.

Matsushima T. 1993. Matsushima Mycological Memoirs No.7. Published by the author, Kobe,

Japan. 75 p.

Matsushima T. 1995. Matsushima Mycological Memoirs No.8. Published by the author, Kobe,

Japan. 54 p.

Ren SC, Ma J, Zhang XG. 2012. Sativumoides and Cladosporiopsis, two new genera of hyphomycetes

from China. Mycologia 11: 443-448. http://dx.doi.org/10.1007/s11557-011-0759-9

Spegazzini C. 1910. Mycetes argentinenses (series V). Anales del Museo Nacional de Historia

Natural Buenos Aires 20: 329-467.

Subramanian CV. 1971. Hyphomycetes. An account of Indian species, expect Cercosporae. Indian

Council of Agricultural Research, New Delhi. 930 p.

Thaung MM. 1979. Two new microfungi from Burma. Transactions of the British Mycological

Society 72: 333-336.

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

Diversity Research Series 15: 1-351.

Xia JW, Ma LG, Castafieda-Ruiz RF, Zhang XG. 2014. Minimelanolocus bicolorata sp. nov.,

Paradendryphiopsis elegans sp. nov. and Corynesporella bannaense sp. nov. from southern

China. Mycoscience 55: 299-307. http://dx.doi.org/10.1016/j.myc.2013.11.003

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

Mycologia 101: 573-578. http://dx.doi.org/10.3852/09-006

MY COTAXON

July-September 2016— Volume 131, pp. 565-574

http://dx.doi.org/10.5248/131.565

Diploschistes tianshanensis sp. nov.,

a corticolous species from Northwestern China

GULIBAHAER ABABAIKELI', ABDULLA ABBAS! “,

SHOU- YU Guo? ®, ANIWAER TUMIER! & REyYIMU MAmutTI!

' Arid Land Lichen Research Center of Western China, College of Life Science & Technology,

Xinjiang University, Urumqi 830046, P. R. China

? State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, PB. R. China

“CORRESPONDENCE TO: “Abdulla@xju.edu.cn ®guosy@im.ac.cn

ABSTRACT—Diploschistes tianshanensis is described as a new species, found on rotten wood

with mosses and other lichens (Cladonia sp.) in an arid region of Northwestern China. The

new fungus, which is diagnosed by urceolate ascomata and large ascospores, resembles

D. gypsaceus but is readily distinguished by its epruinose thallus surface. ITS rDNA sequence

analyses support the taxon as a distinct species. The description of D. tianshanensis is

accompanied by notes on its chemistry, distribution and ecology, and comparison with

related species.

Key worps—Central Asia, biodiversity, Graphidaceae, Ostropales, taxonomy

Introduction

Diploschistes Norman includes crustose lichens with a blackish

pseudoparenchymatous proper exciple, lateral paraphyses, and a trebouxioid

photobiont (Lumbsch 1989, Lumbsch & Mangold 2007). The genus is widely

distributed in arid and semiarid regions worldwide and comprises 35-44 species

(Pérez-Vargas et al. 2012, Fernandez-Brime et al. 2013, Abbas et al. 2014). Most

Diploschistes species occur on rocks, some on soil, and a few species are rarely

found on wood or bark (Lumbsch & Mangold 2007). After Lumbsch (1989)

monographed Holarctic species, several papers on Diploschistes were published

covering cladistic analysis and molecular phylogeny (Lumbsch & Tehler 1998,

Martin et al. 2003, Fernandez-Brime et al. 2013), ecology and distribution

566 ... Ababaikeli & al.

(Pant & Upreti 1993, Guderley & Lumbsch 1996, Lumbsch & Elix 2003,

Mangold et al. 2009), and taxonomy including new species and combinations

(Lumbsch & Elix 1985, 1989; Lumbsch & Mayrhofer 1990; Lumbsch & Aptroot

1993; Lumbsch & Mangold 2007; Pérez-Vargas et al. 2012; Abbas et al. 2014).

The genus exhibits a remarkably variable morphology of the ascomata, which

vary from perithecioid to urceolate and lecanoroid (Lumbsch 1989, Lumbsch

& Mangold 2007). Despite this variation, Diploschistes is currently regarded as

monophyletic and accommodated within the Graphidaceae (Ostropales) based

on molecular studies (Frisch et al. 2006, Martin et al. 2003, Fernandez-Brime

et al. 2013).

The lichen biota of Northwestern China is rich, with more than 670

species (in 127 genera) previously reported (Abbas & Wu 1998, Guo 2005).

Nevertheless, lichen diversity is still incompletely known in the region and new

species can be expected to continue to be described. Of the nine Diploschistes

species recorded from China, four were described from the Northwestern

region (Wei 1991, Guo 2005, Abbas et al. 2014).

An additional Diploschistes species recently collected in Xinjiang,

Northwestern China, is described here as D. tianshanensis, based on

morphological and chemical characters. The ITS region of nrDNA of the type

specimen has also been sequenced and compared with GenBank sequences to

assess the phylogenetic affinities of the new species.

Materials & methods

The specimens studied were collected from Mountain Tianshan, Xinjiang, China,

and deposited in the Herbarium Mycologicum Academiae Sinicae-Lichenes, Beijing,

China (HMAS-L) and the Lichen Section of Botanical Herbarium, Xinjiang University,

Urumgi, China (XJU). Their morphology was examined using a Zeiss Stemi SV 11

stereomicroscope. Sections for anatomical examination were cut by hand using a razor

blade, mounted in water, studied with a Zeiss Axioskop 2 plus light microscope, and

photographed using a Nikon Digital Camera D50. Chemical constituents were identified

by thin-layer chromatography using solvent systems A, B and C (Orange et al. 2010).

DNA EXTRACTION, AMPLIFICATION, AND SEQUENCING. DNA was extracted from

thallus fragments with ascomata of the type specimen using the DNAsecure Plant

DNA Kit (Tiangen, China) following the manufacturer’s protocol. The ITS region was

amplified according to Martin et al. (2003) and Abbas et al. (2014). The entire ITS

region (ITS1+5.8S+ITS2) of the nrDNA repeat tandem was targeted for the Polymerase

Chain Reaction amplification using the primers ITS1 and ITS4 (White et al. 1990)

and performed in a 25uL volume containing 0.75 units of TransStart Taq Polymerase

(Tiangen, China), 2.5 uL of the buffer, 0.5 uL of a 5 uM solution of the primers, 2 uL of

2.5 mM for each dNTP solution, and 1 wL of genomic DNA. Thermocycling protocols

Diploschistes tianshanensis sp. nov. (China) ... 567

were 95°C for 3 min linked to 35 cycles at 94°C for 30 s, 54°C for 30 s, and 72°C for 1

min, with a final extension of 72°C for 10 min. The PCR products were screened on 1%

agarose gels stained with ethidium bromide and sequenced by Genewiz Inc. (Beijing,

China).

Newly obtained sequences were submitted to GenBank. The sequences were

annotated according to Abbas et al. (2014). We excluded the 3’ end of the 18S (SSU)

gene, and the 5’ end of the 28S (LSU) gene from the analyses. Sequences for the new

species were complemented with similar Diploschistes taxa for which ITS sequences

were available in GenBank (TABLE 1). Representative taxa were selected based on

morphological characters, the results of Blast searches of ITS sequence, and the literature

(Martin et al. 2003, Fernandez-Brime et al. 2013, Abbas et al. 2014).

TABLE 1. GenBank accession numbers and voucher information for Diploschistes

tianshanensis and 11 Diploschistes sequences downloaded from GenBank

SPECIES VOUCHER GENBANK

D. tianshanensis 1 China, Abbas 20s1 KC959951

D. tianshanensis 2 China, Abbas 20s2 KC959952

D. cinereocaesius AFTOL-ID 328 HQ650715

D. diacapsis 1 Spain, BCC-Lich 13392 AF228317

D. diacapsis 2 Spain, BCC-Lich 13393 AF228318

D. gypsaceus Switzerland, ESS-9047 AJ458284

D. gyrophoricus Spain, BCC-Lich 11883 AJ458285

D. muscorum 1 Spain, BCC-Lich 13390 AF228319

D, muscorum 2 Spain, BCC-Lich 13391 AF229193

D. rampoddensis Papua New Guinea, hb. Lumbsch AJ458286

D. scruposus Germany, ESS-21508 AJ458287

D. thunbergianus 1 Australia, hb. Lumbsch AJ458289

D. thunbergianus 2 Australia, hb. Lumbsch AJ458290

PHYLOGENETIC ANALYSIS AND SEQUENCE COMPARISONS. The ITS sequences of the

two examined samples and 11 selected representatives were aligned both by ClustalW

and Muscle implemented in MEGA 5 (Tamura et al. 2011). The alignment matrix was

realigned by StatAlign for reliable measurement of the accuracy of the results (Novak et

al. 2008). The final matrix (submitted to TreeBase with accession number $14250) can

be obtained from the corresponding authors.

The evolutionary history was inferred by using the Maximum Likelihood method

(ML) based on the Tamura 3-parameter model in MEGAS and Bayesian inference

(PP) based on GTR model with rates = Invgamma. ‘The analyses involved 13 nucleotide

sequences. Absolute distances were also calculated in MEGAS, using the number of base

differences between sequence pairs and with all gaps removed from each sequence pair.

568 ... Ababaikeli & al.

Results & discussion

Sequence and phylogenetic analysis

The ITS1+5.8S+ITS2 region was successfully sequenced for the two Xinjiang

samples, with the lengths varying by only one base pair (487-488 bp) for the

entire region. The partial sequence containing SSU 3’ end and LSU 5’ end is

included in the data submitted to GenBank.

A few positions in our sample sequences were difficult to align with the

reference sequences and were excluded from the matrix. There were a total of

540 positions in the final dataset, all of which were used in the phylogenetic

analyses.

The ITS sequences indicate that D. tianshanensis probably belongs to the

D. scruposus-group (sensu Martin et al. 2003) with close affinities to the sub-

cosmopolitan species, D. diacapsis (97% identity; 1% gap) and cosmopolitan

species D. gypsaceus (96% identity; 1% gap) and D. scruposus (96% identity;

1% gap). The evolutionary history was inferred as the tree with the highest

log likelihood (-1479.4326). The phylogenetic analyses strongly supported

the monophyly of D. tianshanensis (ML = 100%; PP = 1) and D. diacapsis

(ML = 97%; PP = 1) but only weakly supported (ML = 57%) the relationship

1060/1 1 KC959951 Diploschistes tianshanensis (China, sample 1)

KC959952 Diploschistes tianshanensis (China, sample 2)

AF228317 Diploschistes diacapsis DIADIP-1 (Spain)

AF228318 Diploschistes diacapsis DIADIP-2 (Spain)

AJ458284 Diploschistes gypsaceus ESS 9047 (Switzerland)

AJ458287 Diploschistes scruposus ESS. 21508 (Germany)

AJ458285 Diploschistes gyrophoricus BCC-Lich 11883 (Spain)

AF228319 Diploschistes nuiscorum MUSDIP-1 (Spain)

AF229193 Diploschistes muscorum MUSDIP-2 (Spain)

AJ458289 Diploschistes thunbergianus Eldndge 3800 (Australia)

AJ458290 Diploschistes thunbergianus Lumbsch 10728d (Australia)

HQ650715 Diploschistes cinereocaesius AFTOL-ID 328

AJ458286 Diploschistes rampoddensis Aptroot 39679 (Papua New Guinea)

$7/-

Ficure 1. Phylogenetic relationships inferred from ITS sequences of Diploschistes tianshanensis and

some closely related species in the D. scruposus group (with D. cinereocaesius and D. rampoddensis

as outgroup). Support is indicated for branches with Maximum Likelihood bootstrap frequencies

>50%, and Bayesian Inference posterior probabilities >0.95. The evolutionary tree was inferred by

using the Maximum Likelihood method based on the Tamura 3-parameter model. The tree with

the highest log likelihood (—1479.4326) is shown. ‘The tree is drawn to scale, with branch lengths

measured in the number of substitutions per site.

Diploschistes tianshanensis sp. nov. (China) ... 569

between D. tianshanensis and its morphologically closely related species

including D. diacapsis, D. gypsaceus, and D. scruposus (Fic. 1).

Absolute distances for the aligned sequences of the ITS region also support

the separation of a new species. In our sequence matrix, distances between

infraspecific samples are <3 while distances between species are 213 (TABLE 2).

TABLE 2. Absolute distances between ITS sequences from Diploschistes tianshanensis

and closely related species (gaps ignored in pairwise comparisons).

Infraspecific distances are indicated with bold font.

1 2 3 4 5 6 7 8 9 10

1. D. tianshanensis 1

2. D. tianshanensis 2 1

3. D. diacapsis 1 13 14

4. D. diacapsis 2 15 16 2

5. D. gypsaceus 17 18 17 19

6. D. scruposus 16 17 14 14 15

7. D. gyrophoricus 25 26 24 25 26 24

8. D. muscorum 1 19 20 20 20 22 14 24

9. D. muscorum 2 20 21 21 20 22 14 25 3

10. D. thunbergianus 1 31 32 29 29 35 28 35 26 27

11. D. thunbergianus 2 31 32 29 29 35 28 35 26 27 0

Taxonomy

Diploschistes tianshanensis A. Abbas, S.Y. Guo & Ababaikeli, sp. nov. FIGURE 2

MycoBank MB 804328

Differs from Diploschistes muscorum by its grayish-green thallus without pruina and its

larger ascospores.

Type: China. Xinjiang: Urumqi Co., Tianshan Mountain, Bayi forest farm, 43°21’N

86°49’E, alt. 1760 m, on rotten wood of Picea schrenkiana Fisch. & C.A. Mey., 27 Aug.

2011, A. Abbas 110821 (Holotype, HMAS-L; isotype, XJU; GenBank KC959951,

KC959952).

ErymMo.ocy: The specific epithet tianshanensis refers to the region in which the type

specimen was collected.

THALLUS corticolous, crustose, continuous, whitish-grey to grayish-green,

moderately thin, <0.7 mm thick. Upper surrFace dull, without pruina.

MEDULLA white, amyloid (I+ blue). PHoTroBionT trebouxioid with cells

<16 um diam. PROTHALLUS not visible. VEGETATIVE PROPAGULES absent.

ASCOMATA apothecia, immersed. Disc urceolate, without pruina, orbicular,

0.5-1.0 mm diam. PROPER EXCIPLE dark brown, 60-100 um thick. HyMENIUM

hyaline, 120-180 um high, not inspersed. HyPOTHECIUM yellowish brown,

570 ... Ababaikeli & al.

FiGuRE 2. Diploschistes tianshanensis (holotype, HMAS-L Abbas 110821).

A. habit; B. asci and mature ascospores. Scales: A = 2 mm; B = 20 um.

Diploschistes tianshanensis sp. nov. (China) ... 571

15-30 um thick. PARAPHysES 1-2 um thick, simple, apices not thickened.

Asci cylindrical, 90-130 x 20-30 um, 4-spored. Ascosporss ellipsoid, brown,

muriform, with 4-6 transverse and 1-2 longitudinal septa, 20-36(-42) x 12-18

uum. Pycnipra unknown.

SPOT TESTS—K+ yellow, C+ and KC+ red, PD-.

SECONDARY METABOLITES—Diploschistesic and lecanoric acids detected

GEG):

EcoLocy—At present D. tianshanensis is known only from the type locality

in Northwestern China. It grows on rotten wood of Picea schrenkiana, together

with Cladonia sp. and mosses.

ADDITIONAL SPECIMEN EXAMINED — CHINA. XINJIANG: Urumgi, Tianshan Mountain,

Bayi forest farm, 43°21’N 86°49’E, alt. 1800 m, 15 May 2010, A. Abbas 201000515 (XJU).

ComMENTs —Diploschistes tianshanensis is characterized by its corticolous

habit, grayish green appearance, absence of pruina, urceolate apothecia, large

ascospores, and the presence of diploschistesic and lecanoric acids. To the

best of our present knowledge, other Diploschistes species having a corticolous

habitat include only D. muscorum (Scop.) R. Sant., which can be distinguished

by its combination of 4-spored asci, juvenile parasitism, and the presence of

grayish or whitish pruina. The new species resembles morphologically the

cosmopolitan D. gypsaceus (Ach.) Zahlbr., which differs by its usual occurrence

on calcareous substrata in shaded, damp habitats (Lumbsch 1988, Fletcher &

Hawksworth 2009). Diploschistes gypsaceus has not yet been reported from

China (Wei 1991, Guo 2005, Abbas et al. 2014).

Similar species also include D. diacapsis (Ach.) Lumbsch and D. scruposus

(Schreb.) Norman. Diploschistes diacapsis is a terricolous species that is readily

identified by its whitish pruinose thallus, (4—)8-spored asci, and the ascospore

size (20-40 x 14-20 um). Diploschistes scruposus, which is saxicolous but not

corticolous, also has an epruinose thallus, (4—)8-spored asci, and may or may

not contain diploschistesic acid. A comparison of the diagnostic characters of

D. tianshanensis and morphologically similar species is provided in TABLE 3.

Previous studies that have used DNA sequence data for species recognition

in lichens required both monophyly in single-locus ITS phylogenies and

diagnosable morphological differences (e.g. Han et al. 2013, Abbas et al.

2014). Our morphological and molecular data for our new species meet the

requirement for accurate species delimitation and support recognition of

Diploschistes tianshanensis as a species new to science.

Acknowledgements

The authors thank Dr. Kerry Knudsen (The Herbarium, Department of Botany &

Plant Sciences, University of California, Riverside, California 92521, U.S.A.) and Prof.

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Diploschistes tianshanensis sp. nov. (China) ... 573

Liu-Fu Han (Hebei Normal University, Shijiazhuang 050024, China) for presubmission

review. S.Y. Guo was awarded grants by the National Natural Science Foundation of

China (31370067, 30770012), and A. Abbas was awarded grants by the Ministry of

Science and Technology of China (2012FY111600) and the National Natural Science

Foundation of China (31150003). The valuable assistance given by colleagues is

gratefully acknowledged.

Literature cited

Abbas A, Wu JN. 1998. Lichens of Xinjiang. Sci-Tec & Hygiene Publishing House of Xinjiang,

Urumdi.

Abbas A, Guo SY, Ababaikeli G, Abdulla A, Xahidin H. 2014. Diploschistes xinjiangensis,

a new saxicolous lichen from northwest China. Mycotaxon 129(2): 465-471.

http://dx.doi.org/10.5248/129.465

Fernandez-Brime S, Llimona X, Lutzoni E Gaya E. 2013. Phylogenetic study of Diploschistes

(lichen-forming Ascomycota: Ostropales: Graphidaceae), based on morphological, chemical,

and molecular data. Taxon 62(2): 267-280. http://dx.doi.org/10.12705/622.10

Fletcher A, Hawksworth DL, 2009. Diploschistes. 378-380, in: CW Smith et al. (eds). The Lichens of

Great Britain and Ireland. British Lichen Society, London, pp. 378-380.

Frisch A, Kalb K, Grube M. 2006. Contributions towards a new systematics of the lichen

family Thelotremataceae. Bibliotheca Lichenologica 92. 556 p.

http://dx.doi.org/10.1017/s0024282906219066

Guderley R, Lumbsch HT, 1996. The lichen genus Diploschistes in South Africa (Thelotremataceae).

Mycotaxon 58: 269-292.

Guo SY. 2005. Lichens. 31-82, in: WY Zhuang (ed.). Fungi of northwestern China. Ithaca, New

York, Mycotaxon Ltd.

Han LF, Zhang YY, Guo SY. 2013. Peltigera wulingensis, a new lichen (Ascomycota) from north

China. Lichenologist 45(3): 329-336. http://dx.doi.org/10.1017/s00242829 12000837

Lumbsch HT. 1988. The identity of Diploschistes gypsaceus. Lichenologist 20: 19-24.

http://dx.doi.org/10.1017/s0024282988000052

Lumbsch HT. 1989. Die holarktischen Vertreter der Flechtengattung Diploschistes

(Thelotremataceae). Journal of the Hattori Botanical Laboratory 66: 133-196.

Lumbsch HT, Aptroot A. 1993, Studien tiber die Flechtengattung Diploschistes Il. Nova Hedwigia

56: 237-239.

Lumbsch HT, Elix JA, 1985. A new species of the lichen genus Diploschistes from Australia. Plant

Systematics and Evolution 150: 275-279. http://dx.doi.org/10.1007/bf00984201

Lumbsch HT, Elix JA, 1989. Taxonomy of some Diploschistes spp. (lichenized ascomycetes,

Thelotremataceae) containing gyrophoric acid. Plant Systematics and Evolution 167: 195-199.

http://dx.doi.org/10.1007/bf00936406

Lumbsch HT, Elix JA, 2003. The lichen genus Diploschistes (Thelotremataceae) in Australia.

Bibliotheca Lichenologica 86: 119-128.

Lumbsch HT, Mangold A. 2007. Diploschistes elixii (Ostropales: Thelotremataceae),

an overlooked terricolous species from Western Australia. Lichenologist 39: 459-462.

http://dx.doi.org/10.1017/s0024282907007049

Lumbsch HT, Mayrhofer H, 1990. A new Diploschistes species (lichenized ascomycetes,

Thelotremataceae) from India. Mycotaxon 38: 311-313.

Lumbsch HT, Tehler A, 1998. A cladistic analysis of the genus Diploschistes (Ascomycotina,

Thelotremataceae). Bryologist 101: 398-403. http://dx.doi.org/10.2307/3244178

574 ... Ababaikeli & al.

Mangold A, Elix JA, Lumbsch HT. 2009. Diploschistes. Flora of Australia 57(Lichens 5): 227-239.

Martin MP, Winka PK, Llimona X, Lumbsch HT. 2000. Evaluation of morphological variation in

the lichen Diploschistes ocellatus (Ascomycota, Ostropales): evidence from nuclear rDNA ITS

sequence data. Plant Biology 2(5): 571-578. http://dx.doi.org/10.1055/ s-2000-7471

Martin MP, LaGreca S, Lumbsch HT. 2003. Molecular phylogeny of Diploschistes inferred from ITS

sequence data. Lichenologist 35: 27-32. http://dx.doi.org/10.1006/lich.2002.0427

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Bayesian estimation of alignments and evolutionary trees. Bioinformatics 24(20): 2403-2404.

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edn. British Lichen Society, London.

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33-50. http://dx.doi.org/10.1017/s0024282993000040

Pérez- Vargas I, Hernandez-Padron C, Pérez de Paz PL, Elix JA. 2012. A new saxicolous species

of Diploschistes (Thelotremataceae) from the Canary Islands. Lichenologist 44(1): 67-71.

http://dx.doi.org/10.1017/s0024282911000612

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Wei JC. 1991. An enumeration of lichens in China. International Academic Publishers, Beijing.

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PCR protocols: a guide to methods and applications. San Diego, Academic Press.

http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

MY COTAXON

July-September 2016— Volume 131, pp. 575-581

http://dx.doi.org/10.5248/131.575

Sporidesmiella lushanensis and S. jiulianshanensis spp. nov.

and a new record from China

JIAN Ma*

College of Agronomy, Jiangxi Agricultural University, Nanchang, 330045, China

*CORRESPONDENCE TO: majian821210@163.com

ABSTRACT— Three species of Sporidesmiella were collected on dead branches from subtropical

forests in southern China. Sporidesmiella lushanensis is described as a new species easily

distinguished by its verrucose conidiophores and percurrently extending conidiogenous

cells that produce clavate to cuneiform 3-5-distoseptate conidia with an obviously thickened

dark basal hilum. The new Sporidesmiella jiulianshanensis is distinguished from other

Sporidesmiella species by its ovoid to ellipsoidal smooth conidia with 2 distosepta above and

1 euseptum below. Sporidesmiella garciniae var. brasiliensis is recorded for the first time from

China. These species are described, illustrated, and compared with similar taxa.

KEY worps—anamorphic fungi, asexual morph, hyphomycetes, taxonomy

Introduction

Most of southern China has a humid subtropical climate conducive to

fungal growth, but the anamorphic fungi occurring on plant debris are poorly

known (e.g., Zhang et al. 2009, 2010; Ma et al. 2011, 2012, 2014a,b, 2015;

Ren et al. 2012; Xia et al. 2013). During our continuing surveys (2012-15) of

saprobic microfungi from the forests of southern China, three species referable

to the genus Sporidesmiella P.M. Kirk (Kirk 1982) were collected on dead

branches. Two are new to science while the third represents a new record for

China. Specimens are conserved in the Herbarium of the Department of Plant

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

Sporidesmiella lushanensis Jian Ma, sp. nov. FIG. 1

MycoBank MB 816246

Differs from Sporidesmiella sinensis by its verrucose conidiophores and its wider conidia

with more numerous septa.

576 ... Ma

| 9e9

Ave

1 290

Fic. 1. Sporidesmiella lushanensis (holotype, HJAUP M0239). A, B. Conidiophores, conidiogenous

cells, and conidia; C. Conidiophore; D. Conidia.

Type: China, Jiangxi Province, Lushan Mountain, on dead branches of an unidentified

broadleaf tree, 8 November 2014, J. Ma (holotype, HJAUP M0239).

EryMoLoecy: refers to the locality where the type specimen was found.

Cotonigs effuse on the natural substratum, brown, hairy. Mycelium partly

superficial, partly immersed in the substratum, composed of branched, septate,

pale brown to brown, smooth-walled hyphae. ConipiopHores differentiated,

single, unbranched, erect, straight or flexuous, septate, verrucose, dark brown

to brown, 120-170 um long, 5-6 um wide. CONIDIOGENOUS CELLS integrated,

terminal, cylindrical, verrucose, brown to pale brown, with several enteroblastic

percurrent extensions. Conidial secession schizolytic. Conrp1a holoblastic,

acrogenous, solitary, dry, clavate to cuneiform, pale brown, smooth-walled,

3-5-distoseptate (usually 4), 22-31 um long, 9.5-12 um wide in the broadest

part, base truncate and 3-4.5 um wide, apex rounded.

Sporidesmiella spp. nov. (China) ... 577

TABLE 1. Comparisons of setae, conidiophores, and proliferation

in Sporidesmiella lushanensis and similar species’

SPECIES SETAE CONIDIOPHORES CONIDIOGENOUS CELL PROLIFERATION

S. claviformis' “ Smooth Percurrent

S. latispora’ - Smooth Percurrent

S. lushanensis - Verrucose Percurrent

S. rosae? - Verruculose Percurrent or sympodial

S. setosa* + Smooth Percurrent

S. sinensis? - Smooth Percurrent

*Data from: ! Kirk (1982); ? Wu & Zhuang (2005); * Ma et al. (2012); *McKemy & Wang (1996)

ComMENTS - Sporidesmiella lushanensis resembles S. claviformis P.M. Kirk,

S. latispora W.P. Wu, S. rosae Jian Ma & X.G. Zhang, S. setosa McKemy &

C.J.K. Wang, and S. sinensis W.P. Wu in general conidial shape. However,

S. claviformis can be distinguished by its smooth conidiophores and smaller

conidia with fewer septa; S. latispora differs by its smooth conidiophores

and wider conidia; S. rosae differs by its more numerous conidial septa and

some sympodial proliferation of conidiogenous cells; S. setosa differs by its

smooth conidiophores, longer narrower conidia, and conspicuous setae;

and S. sinensis differs by its smooth conidiophores and narrower conidia

with fewer septa (TABLEs 1, 2).

TABLE 2. Comparisons of conidia in Sporidesmiella lushanensis and similar species’

SPECIES SHAPE SIZE (um) SEPTA BASAL WIDTH (tm)

S. claviformis' Clavate 14-20 x 4.5-9 2 2-2.5

25-30 x

. lati 2 lavate— obovoid 4 5-6

S. latispora Clavate— obovoi 135-17

S. lushanensis Clavate- cuneiform 22-31 x 9.5-12 (3-)4(-5) 3-4.5

4-)5-

S. rosae* Clavate 23-28 x 7-10 re 2.5-3.5

6(-7)

S. setosa* Cuneiform 30-38 x 7.5-9 4-5 3.5-5

S. sinensis? Clavate- cuneiform 24-26 x 7.5-9 3 4

*Data from: ' Kirk (1982); ? Wu & Zhuang (2005); *? Ma et al. (2012); *McKemy & Wang (1996)

—

20um

C E :

—

wiiigz

wun)

Fic. 2. Sporidesmiella jiulianshanensis (holotype, HJAUP M0313). A. Conidiophores showing

conidiogenous cells and percurrent extensions; B, C. Conidiophores and conidia; D, E. Conidia.

Sporidesmiella jiulianshanensis Jian Ma, sp. nov. Fi. 2

MycoBAnk MB 816247

Differs from all other Sporidesmiella species by its ovoid to ellipsoidal conidia with both

eusepta and distosepta.

Type: China, Jiangxi Province, Jiulianshan National Nature Reserve, on dead branches

of an unidentified broadleaf tree, 3 November 2014, J. Ma (holotype, HJAUP M0313).

EryMoLoey: refers to the locality where the type specimen was found.

Cotontgs effuse on the natural substratum, brown, hairy. Mycelium partly

superficial, partly immersed in the substratum, composed of branched, septate,

pale brown to brown, smooth-walled hyphae. Conrp1opHores differentiated,

single, unbranched, erect, straight or flexuous, septate, smooth, brown to

dark brown, 90-205 x 4.5-6 um with up to 5 or more percurrent extensions.

CONIDIOGENOUS CELLS integrated, terminal, cylindrical, brown to pale brown.

Conidial secession schizolytic. Conrp1A holoblastic, acrogenous, solitary, dry,

ovoid to ellipsoidal, brown to pale brown, smooth-walled, 2-distoseptate above

and 1-euseptate below, 21-28 um long, 9.5-10.5 um wide in the broadest part,

apex rounded, base truncate and 2.5-3.5 um wide.

Sporidesmiella spp. nov. (China) ... 579

ComMENtTs - Sporidesmiella cuneiformis (B. Sutton) P.M. Kirk is the only other

species that produces conidia with both eusepta and distosepta. However,

S. cuneiformis has clavate to cuneiform or obovoid, narrower (6-9.5 um)

conidia with (4-)5 distosepta above and 2 eusepta below (Kirk 1982).

Sporidesmiella jiulianshanensis produces somewhat similarly shaped conidia as

S. brachysporioides T.Y. Zhang & W.B. Kendr., which differs by its slightly larger

conidia (15-35 x 7.5-13 um) with 3-4(-5) distosepta (Zhang et al. 1983).

Sporidesmiella garciniae var. brasiliensis Santa Izabel & Gusmio,

Mycosphere 4(2): 158, 2013. FIG. 3

A B

Fic. 3. Sporidesmiella garciniae var. brasiliensis (HJAUP M0250). A, B. Conidiophores and conidia,

with conidiophore apices showing both sympodial and percurrent extensions; C, D. Conidiophores

and conidiogenous cells; E. Conidia.

580 ... Ma

CoONIDIOPHORES differentiated, single, unbranched, smooth, brown to

dark brown, paler at the apex, 140-190 x 3.5-4.5 um. CONIDIOGENOUS CELLS

integrated, terminal, cylindrical, brown to pale brown or subhyaline, with up to

12 or more enteroblastic percurrent extensions, sometimes also with sympodial

extensions. Conidial secession schizolytic. Conrp14 solitary, dry, cylindrical to

subcylindrical, smooth, 4-distoseptate, light brown, 22-31 x 3.5-4.5 um, base

truncate and 1.5-2 um wide, apex rounded.

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

branches of an unidentified broadleaf tree, 8 November 2014, J. Ma (HJAUP M0250).

CoMMENTS - Sporidesmiella garciniae var. brasiliensis, which was originally

described from Brazil (Santa Izabel et al. 2013), has not been previously

recorded in China. It is closely related to S. garciniae Matsush. var. garciniae

in conidial shape but differs in having conidiogenous cells with sympodial

extensions. Our collection corresponds well with the original description of

the Brazilian variety, except that the holotype had slightly narrower conidia

(2.5-4 um) and shorter conidiophores (32-106 um; Santa Izabel et al. 2013).

Acknowledgments

The authors express gratitude to Dr. W.B. Kendrick and Dr. Eric H.C. McKenzie for

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

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

National Natural Science Foundation of China (No. 31360011).

Literature cited

Kirk PM. 1982. New or interesting microfungi. VI. Sporidesmiella gen. nov. (hyphomycetes). Trans.

Br. Mycol. Soc. 79: 479-489. http://dx.doi.org/10.1016/S0007-1536(82)80040-5

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

of a new species. Mycologia 103(2): 407-410. http://dx.doi.org/10.3852/10-176

Ma J, Zhang YD, Ma LG, Castafieda-Ruiz RF, Zhang XG. 2012. Three new species of Sporidesmiella

from southern China. Mycoscience 53: 187-193. http://dx.doi.org/10.1007/s10267-011-0152-1

Ma J, Xia JW, Castafeda-Ruiz RF, Zhang XG. 2014a. Nakataea setulosa sp. nov. and

Uberispora formosa sp. nov. from southern China. Mycol. Progress 13: 753-758.

http://dx.doi.org/10.1007/s11557-013-0958-7

Ma J, Zhang XG, Castafieda-Ruiz RF. 2014b. Ceratosporium hainanensis and Solicorynespora

obovoidea spp. nov., and a first record of Bactrodesmiastrum obscurum from southern China.

Mycotaxon 127: 135-143. http://dx.doi.org/10.5248/127.135

MaJ, Xia JW, Castaneda-Ruiz RF, Zhang XG. 2015. Two new species of Sporidesmiella from southern

China. Nova Hedwigia 101: 131-137. http://dx.doi.org/10.1127/nova_hedwigia/2015/0254

McKemy JM, Wang CJK. 1996. A new species of Sporidesmiella from New York. Mycologia 88:

129-131. http://dx.doi.org/10.2307/3760792

Ren SC, Ma J, Ma LG, Zhang YD, Zhang XG. 2012. Sativumoides and Cladosporiopsis, two new

genera of hyphomycetes from China. Mycol. Progress 11: 443-448.

http://dx.doi.org/10.1007/s11557-011-0759-9

Sporidesmiella spp. nov. (China) ... 581

Santa Izabel TS, Cruz ACR, Gusmao LFP. 2013. Conidial fungi from the semi-arid Caatinga biome

of Brazil. Ellisembiopsis gen. nov., new variety of Sporidesmiella and some notes of Sporidesmium

complex. Mycosphere 4(2): 156-163. http://dx.doi.org/10.5943/mycosphere/4/2/1

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

Divers. Res. Ser. 15. 351 p.

Xia JW, Ma LG, Ma J, Zhang XG. 2013. Two new species of Spadicoides from southern China.

Mycotaxon 126: 55-60. http://dx.doi.org/10.5248/126.55

Zhang TY, Kendrick B, Brubacher D. 1983. Annellidic (percurrent) and sympodial proliferation in

congeneric hyphomycetes, and a new species of Sporidesmiella. Mycotaxon 18: 243-257.

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

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

Zhang YD, Ma J, Ma LG, Zhang XG. 2010. A new species of Heteroconium from Fujian, China.

Mycotaxon 114: 315-318. http://dx.doi.org/10.5248/114.315

MY COTAXON

July-September 2016— Volume 131, pp. 583-588

http://dx.doi.org/10.5248/131.583

Corynesporopsis obclavata and Stanjehughesia jiangxiensis spp. nov.

from Lushan Mountain, China

JIAN Ma*

College of Agronomy, Jiangxi Agricultural University, Nanchang, 330045, China

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

ABSTRACT—Two new species, Corynesporopsis obclavata and Stanjehughesia jiangxiensis,

are described and illustrated from decaying bamboo culms collected in Lushan Mountain,

China. Corynesporopsis obclavata is characterized by its obclavate, smooth, 5-9-euseptate

conidia. Stanjehughesia jiangxiensis is easily distinguished by its obclavate to obclavate-

rostrate, smooth, 11-14-euseptate conidia.

KEY worps—anamorphic fungi, bambusicolous fungi, hyphomycetes, taxonomy

Introduction

Bamboos are known to host a number of highly characteristic fungi, of

which over 1100 species have been described or recorded (Hyde et al. 2002).

Nonetheless, our knowledge of bamboo fungi is incomplete and still at the

cataloguing stage (Hyde et al. 2002). To enrich knowledge of bambusicolous

fungi, we have been investigating saprobic microfungi occurring on decaying

culms of bamboo. Collecting in the forest ecosystems of Lushan Mountain

yielded two new hyphomycetes that clearly belong to the genera Corynesporopsis

PM. Kirk (Kirk 198la) and Stanjehughesia Subram. (Subramanian 1992).

The new species are described, illustrated, and compared with similar taxa.

Specimens are conserved in the Herbarium of the Department of Plant

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

Corynesporopsis obclavata Jian Ma, sp. nov. FIG. 1

MycoBank MB 816248

Differs from Corynesporopsis rionensis by its much longer, obclavate conidia with more

numerous eusepta.

584 ... Ma

20um

Fic. 1. Corynesporopsis obclavata (holotype, HJAUP M0204).

A, B. Conidiophores with catenate conidia; C. Conidiophores; D. Conidia.

Type: China, Jiangxi Province, Lushan Mountain, on decaying culms of bamboo,

8 November 2014, J. Ma (Holotype, HJAUP M0204).

ETYMOLOGy: refers to the obclavate conidia.

Corynesporopsis & Stanjehughesia spp. nov. (China) ... 585

COLONIES on natural substrate effuse, dark brown to black, hairy. Mycelium

partly superficial, partly immersed in the substratum, composed of branched,

septate, pale brown to brown, smooth hyphae. ConrpioPpHor:s differentiated,

single, erect, unbranched, straight or flexuous, brown to dark brown, smooth,

septate, 100-135 x 5-7 um. CONIDIOGENOUS CELLS monotretic, integrated,

terminal, determinate, sometimes percurrent, cylindrical, brown, smooth,

17-22 x 5-6 um. Conidial secession schizolytic. CoNnmpIA acrogenous,

catenate, in chains of two, obclavate, brown, paler toward the apex, smooth,

5-9-euseptate, 27-61 tm long, 6-12 um diam. in the broadest part, protruding

a 2.5-4 um diam. basal scar.

COMMENTS—Corynesporopsis, established by Kirk (1981la) with Corynespora

quercicola Borowska [= Corynesporopsis quercicola (Borowska) P.M. Kirk] as

type species, is characterized by acrogenous, shortly catenate, euseptate conidia

arising from terminal, integrated, monotretic, determinate or rarely percurrently

extending conidiogenous cells on differentiated conidiophores. Fifteen species

have been described or transferred to the genus based on differences in conidial

morphology including shape, size, septation, ornamentation, and pigmentation

(Kirk 1981a,b, 1983; Holubova-Jechova & Mercado Sierra 1986; Holubova-

Jechova 1987; Morgan-Jones 1988; Sutton 1989; Castafeda-Ruiz & Kendrick

1990; Matsushima 1993; Castafieda-Ruiz et al. 2010; Ma et al. 2012; Xia et

al. 2013, 2014; Hernandez-Restrepo et al. 2014; Kirschner 2015). Castafieda-

Ruiz et al. (2010) provided a key and illustrations to the ten species known at

that time; Hernandez-Restrepo et al. (2014) provided an expanded key that

included two additional species; and Xia et al. (2014) provided a synoptic table

of the main morphological features that distinguished 13 accepted species.

Corynesporopsis obclavata is similar only to C. rionensis Hol.-Jech., which

is clearly differentiated by its fusiform to ellipsoidal, 24-36 x 8-11 um,

3-4(-5)-euseptate conidia (Holubova-Jechova & Mercado Sierra 1986).

Stanjehughesia jiangxiensis Jian Ma, sp. nov. Fic. 2

MycoBAank MB 816249

Differs from Stanjehughesia fasciculata by its wider, smooth conidia with eusepta but

no distosepta; from S. fusiformis by its larger conidia with more numerous eusepta; and

from S. polypora by its shorter conidia with fewer eusepta and without a germ pore in

each cell.

Type: China, Jiangxi Province, Lushan Mountain, on decaying culms of bamboo,

8 November 2014, J. Ma (Holotype, HJAUP M0274).

EryMoLoey: refers to the province where the type specimen was found.

COLONIES on natural substrate effuse, brown. Mycelium superficial,

composed of branched, septate, pale brown to brown, smooth-walled hyphae.

586 ... Ma

Fic. 2. Stanjehughesia jiangxiensis (holotype, HHAUP M0274).

Conidiogenous cells and conidia.

CONIDIOPHORES reduced to conidiogenous cells. CONIDIOGENOUS CELLS

monoblastic, determinate, solitary or grouped, cylindrical or ampulliform,

brown to dark brown, smooth, 9.5-22 x 3.5-4 um. Conidial secession

schizolytic. Conrp14_ holoblastic, solitary, acrogenous, straight or curved,

obclavate to obclavate-rostrate, brown, smooth, 11-14-euseptate, 82-120 um

long, 10-12 um diam. in the broadest part, tapering to 2.5-3 um near the apex,

3.5-4 um diam. at the truncate base.

COMMENTS—Subramanian (1992) established Stanjehughesia to accommodate

five Sporidesmium species, including S. hormiscioides Corda [= Stanjehughesia

hormiscioides (Corda) Subram.] as type species. The genus is mainly

Corynesporopsis & Stanjehughesia spp. nov. (China) ... 587

characterized by very reduced or absent conidiophores and solitary, euseptate

conidia seceding schizolytically from monoblastic, determinate, lageniform

or ampulliform, short conidiogenous cells (Subramanian 1992, Wu & Zhuang

2005, Seifert et al. 2011). Sixteen taxa have been included in Stanjehughesia

(Subramanian 1992, McKenzie 1995, Mena-Portales et al. 2001, Wu & Zhuang

2005, Delgado 2008, Marincowitz et al. 2008, Ma et al. 2011, Almeida et al.

2014). However, only 14 species are currently accepted, of which seven have

been recorded from China (Lu et al. 2000, Wu & Zhuang 2005, Ma et al. 2011).

Stanjehughesia jiangxiensis is similar to S. fasciculata J. Mena et al.,

S. fusiformis W.P. Wu, and S. polypora W.P. Wu in conidial shape. However,

S. fasciculata differs by its narrower (7-11 um) verruculose conidia with both

eusepta and distosepta and with a broken outer wall at the apical region (Mena-

Portales et al. 2001); S. fusiformis differs by its much smaller (65-85 x 7-8 um)

6-8-euseptate conidia with a rostrate subhyaline apex and a wider (4-5 um)

base (Wu & Zhuang 2005); and S. polypora differs by its longer (110-165 um)

15-20-euseptate conidia with a germ pore in each cell (Wu & Zhuang 2005).

Acknowledgments

The authors express gratitude to Dr. W.B. Kendrick and Dr. Eric H.C. McKenzie for

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

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

National Natural Science Foundation of China (No. 31360011).

Literature cited

Almeida DAC, Miller AN, Gusmao LFP. 2014. New species and combinations of conidial

fungi from the semi-arid Caatinga biome of Brazil. Nova Hedwigia 98: 431-447.

http://dx.doi.org/10.1127/0029-5035/2013/0162

Castaneda-Ruiz RF, Kendrick B. 1990. Conidial fungi from Cuba: II. Univ. Waterloo Biol. Ser. 33.

61 p.

Castafieda-Ruiz RF, Silvera-Simon C, Gené J, Guarro J, Minter DM, Stadler M, Saikawa M.

2010. A new species of Corynesporopsis from Portugal. Mycotaxon 114: 407-415.

http://dx.doi.org/10.5248/114.407

Delgado G. 2008. South Florida microfungi: a new species of Stanjehughesia (hyphomycetes) from

Sabal palm. Mycotaxon 103: 229-234.

Hernandez-Restrepo M, Gené J, Castafteda-Ruiz RE, Kirk PM, Guarro J. 2014. A new species of

Corynesporopsis from Spain. Mycotaxon 127: 155-160. http://dx.doi.org/10.5248/127.155

Holubova-Jechova V. 1987. Studies on hyphomycetes from Cuba VI. New and rare species with

tretic and phialidic conidiogenous cells. Ceska Mykol. 41(2): 107-114.

Holubova-Jechova V, Mercado Sierra A. 1986. Studies on hyphomycetes from Cuba IV.

Dematiaceous hyphomycetes from the Province Pinar del Rio. Ceska Mykol. 40(3): 142-164.

Hyde KD, Zhou DQ, Dalisay T. 2002. Bambusicolous fungi: a review. Fungal Divers. 9: 1-14.

Kirk PM. 1981a. New or interesting microfungi II. Dematiaceous hyphomycetes from Esher

Common, Surrey. Trans. Br. Mycol. Soc. 77(2): 279-297.

http://dx.doi.org/10.1016/S0007-1536(81)80031-9

588 ... Ma

Kirk PM. 1981b. New or interesting microfungi HI. A preliminary account of microfungi

colonizing Laurus nobilis leaf litter. Trans. Br. Mycol. Soc. 77(3): 457-473.

http://dx.doi.org/10.1016/S0007-1536(81)80093-9

Kirk PM. 1983. New or interesting microfungi VIII. Corynesporopsis indica sp. nov.. Mycotaxon

17: 405-408.

Kirschner R. 2015. Phylogenetic placement of a new species of Corynesporopsis from dead acacia

wood indicates occurrence of tretic conidiogenesis within Xylariales. Phytotaxa 192 (1): 24-34.

http://dx.doi.org/10.11646/phytotaxa.192.1.3

Lu BH, Hyde KD, Ho WH, Tsui K., Taylor JE, Wong KM, Yanna, Zhou DQ. 2000. Checklist of Hong

Kong Fungi. Fungal Divers. Res. Ser. 5. 207 p.

Ma J, Ma LG, Zhang YD, Zhang XG. 2011. Three new hyphomycetes from southern China.

Mycotaxon 117: 247-253. http://dx.doi.org/10.5248/117.247

Ma J, Ma LG, Zhang YD, Castafieda-Ruiz RF, Zhang XG. 2012. New species and record of

Corynesporopsis and Hemicorynespora from southern China. Nova Hedwigia 95: 233-241.

http://dx.doi.org/10.1127/0029-5035/2012/0030

Marincowitz S, Crous PW, Groenewald JZ, Wingfield MJ. 2008. Microfungi occurring on the

Proteaceae in the fynbos. CBS Biodivers. Ser. 7. 166 p.

Matsushima T. 1993. Matsushima Mycological Memoirs No. 7. Published by the author, Kobe,

Japan.

McKenzie EHC. 1995. Dematiaceous hyphomycetes on Pandanaceae. 5. Sporidesmium sensu lato.

Mycotaxon 56: 9-29.

Mena-Portales J, Delgado-Rodriguez G, Mercado-Sierra A, Gené J, Guarro J, Iacona V. 2001. New

or interesting hyphomycetes from the Biosphere Reserve of Sierra del Rosario, Cuba. Mycologia

93: 751-757. http://dx.doi.org/10.2307/3761830

Morgan-Jones G. 1988. Notes on hyphomycetes. LVII. Corynespora biseptata, reclassified in

Corynesporopsis. Mycotaxon 31(2): 511-515.

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

Biodiversity Series 9. 997 p.

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

Divers. Res. Ser. 15. 351 p.

Xia JW, Ma LG, Ma YR, Castafieda-Ruiz RE, Zhang XG. 2013. Corynesporopsis curvularioides

sp. nov. and new records of microfungi from southern China. Cryptogamie, Mycol. 34(3):

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66(2): 241-248. http://dx.doi.org/10.12905/0380.sydowia66(2)2014-0241

MY COTAXON

July-September 2016— Volume 131, pp.589-596

http://dx.doi.org/10.5248/131.589

New records of Rhizocarpon from Hengduan Mountains, China

WEI-CHENG WANG!, ZHAO-JIE REN’, LU-LU ZHANG’, & ZUN-TIAN ZHAO?”

'Key Laboratory of Systematic Mycology and Lichenology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100101, China

*Natural Department, Shandong Museum, Jinan 250014, China

College of Life Sciences, Shandong Normal University, Jinan 250014, China

* CORRESPONDENCE TO: ztzhao@sohu.com

ABSTRACT—Four species of lichen-forming fungi from Hengduan Mountains are reported

new to Asia—Rhizocarpon effiguratum, R. intersitum, R. subgeminatum, and R. umbilicatum.

These species are described and illustrated in detail and comments on their ecology and

distribution ranges are provided.

Key worps—Rhizocarpaceae, Asia, taxonomy

Introduction

The Hengduan Mountains is a large mountainous region in southwest China

(22°-32°N 97°-103°E) and occupies parts of Sichuan, Xizang, and Yunnan

Provinces. It consists of several mountain ranges, most of which run roughly

north to south, and elevations span 4000-6000 m. Most mountain peaks are

permanently covered with snow. Three great rivers of China, the Yangtze,

Mekong, and Salween, run in deep parallel valleys separated by mountain

ranges. Vast stretches of stones that serve as substrates for various lichens are

common in this montane region (FIG. 1).

The Hengduan Mountains were chosen for study because of their rich

lichen flora. There is a surprising lack of published information concerning the

microlichens of this area. Rhizocarpon is a particularly under-studied genus,

with only a few species previously recorded: R. alpicola, R. geographicum,

R. lavatum, R. rubescens, and R. sinense (Wang et al. 2015, Zahlbruckner

1930, Zhao et al. 2013). Here we report four additional Rhizocarpon species

590 ... Wang & al.

Fic. 1. The ecotope of Mt. Zheduo, located in Kangding Co., showing the typically rocky landscape

in the Hengduan Mountains.

(R. effiguratum, R. intersitum, R. subgeminatum, R. umbilicatum) collected in

the Hengduan Mountains as new to China as well as to Asia.

Materials & methods

Observations are based on material collected by the authors from

Hengduan Mountains in 2015 and preserved in the herbarium of Shandong

Normal University, Jinan, China (SDNU), and specimens held in the lichen

herbarium of Kunming Institute of Botany, Chinese Academy of Sciences,

Kunming, China (KUN-L). Morphological and anatomical characters were

examined under an Olympus SZ stereomicroscope and an Olympus CX21

polarizing microscope. Both thallus and medulla were tested with K (10%

aqueous solution of potassium hydroxide), C (saturated solution of aqueous

sodium hypochlorite), I (10% aqueous solution of aqueous potassium iodide),

and P (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).

Lichen photographs were taken with an Olympus DP72 camera attached to the

Olympus SZX16 and BX61 microscopes.

Rhizocarpon spp. new for Hengduan region (China) ... 591

ees:

we,

ocd

ash

ats 3

ts ae -

A wee ee hed a

nin Oy, eee GL Gabe a OS

10 pm

Fic. 2. Rhizocarpon effiguratum (SDNU Wang 20150590). A: habitus, distinct black prothallus

along the margin of the thallus; B: apothecium section with brown-black epihymenium and poorly

developed exciple; C: 1-septate, brown, halonate ascospores. Scale bars: A = 2 mm; B = 50 um;

C= 10 um.

Taxonomic descriptions

Rhizocarpon effiguratum (Anzi) Th. Fr., Lich. Scand. 1(2): 613 (1874) Fic. 2

Saxicolous. THALLUs bright yellow, areolate, <3.0 cm diam; medulla

I+ violet; prothallus well developed, black, visible between the areolae and

along the margin; areoles convex, matt, contiguous, round, 0.2-0.5(-0.8) mm

diam. APOTHECIA 0.25-0.8 mm diam between the areolae, black, epruinose,

angular, flat to convex, scabrid, margin persistent; exciple poorly developed;

epihymenium green-black or brown-black, hymenium colorless. Asci1 8-spored,

Rhizocarpon-type; ascospores ellipsoid, 1-septate, dark green or brown, paler

when young, halonate, 12-15 x 5-6 um (n = 120).

592 ... Wang & al.

CHEMISTRY—Medulla K-, C-, P+ yellow; rhizocarpic and psoromic acids

detected by TLC.

SPECIMENS EXAMINED: CHINA. X1zANG, Chayu Co., Demula col, 29°19.15’N

97°01.75’E, 4794 m a.s.l., on siliceous rock, 23 Sep. 2014, Li S. Wang et al. 14-46628

(KUN-L); Zuogong Co., Xinde vil., 29°43.57’N 98°04.80’E, 4730 m a.s.l., on siliceous

rock, 16 Sep. 2014, Li S. Wang et al. 14-45653, 14-45661 (KUN-L); Mt. Dongda, 5200

m a.s.l., on siliceous rock, 20 Aug. 2004, Li S. Wang 04-25614 (KUN-L); along the road

from Mangkang to Zuogong, 4742 m a.s.l., on siliceous rock, 15 Sep. 2014, Li S. Wang et

al. 14-45562, 14-45574 (KUN-L); Mangkang Co., Dogdala col, 29°05.48’N 98°36.94’E,

4930 ma.s.l., 17 Aug. 2007, Li S. Wang et al. 07-27990 (KUN-L). SICHUAN: Ganzi City,

Shiqu Co., Luoxv vil., Zebuka col, 4900 m a.s.l., on siliceous rock, 9 Sep. 2009, Li S. Wang

09-30462 (KUN-L); Litang Co., Mt. Haizishan, 29°41’32.79”N, 100°23’20.18”E, 4600 m

a.s.l., on siliceous rock, 17 Oct. 2015, Wei C. Wang 20150591, 20150589, 20150593,

20150598, 20150599, 20150600 (SDNU); Litang Co., Mt. Tuwershan, 29°31’00”N

100°16’11”E, 4896 m a.s.l., on siliceous rock, 17 Oct. 2015, Wei C. Wang 20150590,

20150592, 20150594, 20150595, 20150596, 20150597, 20150603, 20150604 (SDNU).

DISTRIBUTION—Rhizocarpon effiguratum has been previously reported from

Europe and North America (Anderson 1965, Timdal & Holtan-Hartwig 1988,

Nimis 1993, Llimona & Hladun 2001). New to Asia.

ComMENTS—Rhizocarpon effiguratum is a small lichen found in China on

high mountains above 4000 m a.s.l. It is characterized by its yellow thallus with

distinct black prothallus, I+ violet medulla, angular black apothecia, small

brown 1-septate ascospores, and the presence of psoromic acid. Rhizocarpon

pusillum Runemark and R. parvum Runemark are similar to R. effiguratum with

small, pigmented, 1-septate ascospores, but both are lichenicolous and have a

much smaller thallus (<1 cm diam). Rhizocarpon superficiale (Schaer.) Malme,

which also resembles R. effiguratum, is distinguished by its larger (26 cm diam)

thallus, and different lichen substances: R. superficiale contains rhizocarpic,

stictic, and hypostictic acids whereas R. effiguratum has rhizocarpic and

psoromic acids. Rhizocarpon alpicola (Wahlenb.) Rabh. and R. eupetraeoides

(Nyl.) Blomb. & Forssell have longer (218 um) ascospores; Zahlbruckner

(1930) reported the only known Chinese occurrence of R. alpicola from

Yunnan Province. Rhizocarpon norvegicum Rasanen, another species similar

to R. effiguratum, also produces brown 1-septate ascospores and an I+ violet

medulla, but differs in its scattered thallus; it is known from Europe and North

America (Thomson 1997).

Rhizocarpon intersitum Arnold, Verh. Zool.-Bot. Ges. Wien 27: 554 (1877) Fic. 3

Saxicolous. THALLUs areolate, 2-3 cm diam; medulla I-; prothallus distinct,

black; areoles dark grey, slightly to strongly convex. ApoTHeciA black,

epruinose, between the areoles, closely attached with areoles, up to 0.2-1.0 mm

diam, flat to convex, margin present; exciple poorly developed, epihymenium

Rhizocarpon spp. new for Hengduan region (China) ... 593

Fic. 3. Rhizocarpon intersitum (SDNU Wang 20150601). A: habitus; B: submuriform green

halonate ascospores. Scale bars: A = 1 mm; B = 10 um.

olive-green, K-; hymenium hyaline, upper part with green tinge, thick, =>100

um tall. Ascr 8-spored; ascospores ellipsoid, submuriform, with 4-7 cells in

optical view, dark green, halonate, 20-25 x 12 um (n = 120).

CHEMISTRY—Medulla K -, C -, P -. No lichen substances detected by TLC.

SPECIMENS EXAMINED: CHINA. SICHUAN, Xiangcheng Co., Mt. Daxueshan, 28°34’N

99°49’E, 4650 m a.s.l., on siliceous rock, 12 Sep. 2002, Li S. Wang 02-21488, 02-22354

(KUN-L); Luding Co., Mt. Gongga, Yajiagen, 29°54’36”N 102°00’14’E, 3800 ma.s.l., on

siliceous rock, 13 Oct. 2015, Wei C. Wang 20150574 (SDNU). YUNNAN, Jianchuan Co.,

Mt. Laojun, 26°38’12”N 99°47’08”E, 4000 m a.s.l., on siliceous rock, 25 Aug. 2015, Wei

C. Wang 20150601 (SDNU).

DISTRIBUTION—Rhizocarpon intersitum has been previously reported from

North America and Scandinavia (Anderson 1965, Timdal & Holtan-Hartwig

1988). New to Asia.

ComMENTS—Rhizocarpon intersitum is characterized by its dark 4-7-celled

ascospores, an olive-green epihymenium, a distinct black prothallus, a dark

grey thallus, and absence of lichen substances. Rhizocarpon flavum C.W. Dodge

& G.E. Baker is close to R. intersitum but has a whitish or yellow to green-

yellow thallus, a dark brown K+ purple epihymenium, and 2-6 ascospores per

ascus.

Rhizocarpon subgeminatum Eitner, Jahresber. Schles. Ges. Vaterl. Cult. 88: 42

(1911) Fic. 4

Saxicolous. THALLUS crustose, dark grey; medulla I-; prothallus black;

areoles 0.2-0.7 mm diam, flat to slightly to convex, angular. APOTHECIA

594 ... Wang & al.

Fic. 4. Rhizocarpon subgeminatum (SDNU Wang 20150609). A: habitus; B: apothecium section;

C: crystals in exciple (the sample was in water); D: two spores per ascus; E: muriform ascospores.

Scale bars: A = 2 mm; B, C = 50 um; D, E= 10 um.

black, 0.5-0.8 mm diam, flat, between areoles, scabrid, epruinose; exciple well

developed, black at the rim, inner part red-brown, containing crystals, extend to

the bottom of hypothecium, <100 um wide, K+ red; epihymenium red-brown,

K+ red-purple (Atra-red); hymenium usually with pale green tinge, thick,

about 125 um tall. Ascr 2-spored; ascospores ellipsoid, muriform, colorless,

halonate, 37-55 x 20-25 um (n = 30).

CHEMISTRY—Medulla K-, C-, P-. Barbatic acid detected by TLC.

SPECIMENS EXAMINED: CHINA. SicHuaNn, Luding Co. Mt. Gongga, Yajiagen,

29°54’36’"N 102°00'14”E, 3800 m a.s.l., on siliceous rock, 13 Oct. 2015, Wei C. Wang

20150605, 20150606, 20150607, 20150608 (SDNU); Kangding Co., Mt. Zheduo,

30°04’29’N 101°48’12”E, 4200 m a.s.l., on siliceous rock, 15 Oct. 2015, Wei C. Wang

20150609 (SDNU).

DISTRIBUTION—Rhizocarpon subgeminatum has been previously reported

from Europe and North America (Timdal & Holtan-Hartwig 1988, Fletcher et

al. 2009). New to Asia.

ComMENtTs—Ihlen (2004), who reported a chemotype without lichen

substances, suggested that one specimen containing barbatic acid was probably

Rhizocarpon spp. new for Hengduan region (China) ... 595

contaminated by fragments of R. rittokense. However, the R. subgeminatum

specimens collected in southwest China all contain barbatic acid detected by

TLC, strongly supporting R. subgeminatum as having two chemotypes: 1) one

with barbatic acid, and 2) one with no lichen substances.

Rhizocarpon umbilicatum (Ramond) Flagey, Mém. Soc. Emu. Doubs, sér. 6, 8: 98

(1894) FIG. 5

Saxicolous. THALLUs rimose to areolate, white, pruinose, thick, 0.5-1.0 mm

tall; medulla I-; prothallus present only along the margin. APoTHeEcza black,

immersed, epruinose, 0.5-1.5 mm diam, disc flat, margin distinct, usually with

a white pruina. Exciple poorly developed; epihymenium green-brown K-,

containing crystals; hymenium colorless, thin, about 120 um tall, containing

crystals. Asci 8-spored; ascospores ellipsoid, submuriform, with 6-10 cells in

optical view, colorless, halonate, 25-30 x 15-18 um (n = 30).

Fic. 5. Rhizocarpon umbilicatum (KUN-L Wang 94-15489). A: white thallus and apothecia

with pruinose margin; B: submuriform halonate ascospores; C: apothecium section; D: crystals

in epihymenium and hymenium (the sample was in water). Scale bars: A = 1 mm; B = 10 um;

C, D = 100 um.

596 ... Wang & al.

CHEMISTRY—Medulla K+ yellow, C-, P+ orange. Stictic acid detected by

ThE.

SPECIMENS EXAMINED: CHINA. SICHUAN, Yanyuan Co., Mt. Huolu, 4150 m a.s.l., on

siliceous rock, 23 Jul. 1983, Li S. Wang 83-1199 (KUN-L); YUNNAN, Deqin Co., Mt.

Baimaxueshan, 4200 m a.s.l., on siliceous rock, 3 Oct. 1994, Li S. Wang 94-15489

(KUN-L).

DISTRIBUTION—Rhizocarpon umbilicatum has been previously reported from

Europe, North America, and Australia (Fletcher et al. 2009). New to Asia.

CoMMENTS—Rhizocarpon umbilicatum is close to R. petraeum (Wulfen)

A. Massal., which differs by its longer (230 um) ascospores and apothecia

lacking a pruinose margin. Rhizocarpon umbilicatum is characterized by

its white pruinose thallus, apothecia with pruinose margin, submuriform

ascospores, and presence of stictic acid.

Acknowledgements

We thank Dr. Martin Westberg (Swedish Museum of Natural History, Stockholm,

Sweden) and Dr. S. Pérez-Ortega (Royal Botanical Garden, Madrid, Spain) for

presubmission reviews; we also thank E. Timdal (University of Oslo, Norway) for helpful

suggestions during our research. This study was supported by the National Natural

Science Foundation of China (31170187, 31400015, 31570017), and the Scientific

Research Innovation Foundation of Shandong Normal University (SCX1525).

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Peninsula and Balearic Islands. Bocconea 14. 581 p.

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Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2nd

edition. London: British Lichen Society.

Thomson JW. 1997. American Arctic lichens 2. The microlichens. Madison WI, University of

Wisconsin Press. 675 p.

Timdal E, Holtan-Hartwig J. 1988. A preliminary key to Rhizocarpon in Scandinavia. Graphis

Scripta 2: 41-54.

Wang WC, Zhao ZT, Zhang LL. 2015. Four Rhizocarpon species new to China. Mycotaxon 130:

883-891. http://dx.doi.org/10.5248/130.883

Zahlbruckner A. 1930. Lichenes. Symbolae Sinicae 3. 254 p.

Zhao ZT, Li C, Zhao X, Zhang LL. 2013. New records of Rhizocarpon from China. Mycotaxon 125:

217-226. http://dx.doi.org/10.5248/125.217

MY COTAXON

July-September 2016— Volume 131, pp. 597-603

http://dx.doi.org/10.5248/131.597

Ellisembia henanensis sp. nov. and

two new hyphomycete records from central China

JI-WEN X1A, JIN-YE WANG, CHUN-LING YANG,

ZHUANG LI* & XIU-GUO ZHANG ®

Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong, 271018, China

* CORRESPONDENCE TO: “/iz552@126.com, * sdau613@163.com

ABSTRACT—Ellisembia henanensis is described and illustrated from specimens collected on

dead branches in Henan Province. The fungus is characterized by distinct single unbranched

erect conidiophores, monoblastic conidiogenous cells, and distoseptate fusiform to ellipsoidal

conidia. Lylea tetracoila and Menispora glauca are newly recorded from China.

Key worps—conidial fungi, taxonomy

Introduction

During our ongoing survey of microfungi associated with woody debris in

subtropical forests of Henan provinces, three species with the morphological

characteristics of the genera Ellisembia Subram., Lylea Morgan-Jones, and

Menispora Pers. (Subramanian 1992, Morgan-Jones 1975, Persoon 1822)

were collected on decaying twigs and dead stems. One of these represents an

undescribed species of Ellisembia, while the other two specimens represent new

species for China. Specimens are deposited in the Herbarium of Department

of Plant Pathology, Shandong Agricultural University, Taian, Shandong, China

(HSAUP) and the Mycological Herbarium, Institute of Microbiology, Chinese

Academy of Sciences, Beijing, China (HMAS).

Materials & methods

Samples of woody debris collected were placed in separate zip-lock plastic bags,

taken to the laboratory, and then incubated at 27°C for more than 2 weeks in an artificial

598 ... Xia & al.

climate box in 9 cm diameter plastic Petri dishes containing moistened filter paper.

Samples were examined under an Olympus SZ61 dissecting microscope. Fifty mature

conidia and 30 conidiophores were mounted in lactophenol, measured at 600x and

1000x magnifications, and photographed with an Olympus BX53 microscope. Adobe

Photoshop 7.0 was used to prepare the photographic images; the backgrounds replaced

for esthetic reasons.

Taxonomy

Ellisembia henanensis J.W. Xia & X.G. Zhang, sp. nov. FIG. 1

MycoBank MB 818289

Differs from Ellisembia ellipsoidea by its smaller conidia with fewer distosepta.

Type: China, Henan Province, Jigong Mountain, on dead stems of an unidentified

broadleaf tree, 16 Oct. 2015, J.W. Xia (Holotype, HSAUP H6697; isotype, HMAS

245597).

EryMoLoGey: in reference to the type locality.

COLONIES on natural substrate effuse, pale brown to brown, hairy. Mycelium

partly superficial, partly immersed in the substrate, composed of septate, pale

brown, smooth, 2-4 um wide hyphae. ConrpropHores differentiated, single,

erect, straight to slightly flexuous, cylindrical, smooth, thick-walled, pale brown

to brown, 3-5-septate, 25-50 x 4-5 um. CONIDIOGENOUS CELLS monoblastic,

integrated, terminal, pale brown to brown, 10-15 x 4-5 um. Conidial secession

schizolytic. Conip14 solitary, acrogenous, straight or slightly curved, fusiform

to ellipsoidal, smooth-walled, pale brown to brown, 4-5-distoseptate, 25-35 x

6-7.5 um, 1.5-2 um wide at the truncate base.

ComMMENTS—Ellisembia was introduced by Subramanian (1992) to

accommodate Sporidesmium-like species with determinate or irregularly

percurrently extending conidiogenous cells that produce distoseptate conidia.

Wu & Zhuang (2005) merged Imicles Shoemaker & Hambl. (Shoemaker &

Hambleton 2001) into Ellisembia and expanded the generic concept to include

species that produce typically lageniform, ovoid, or doliiform percurrently

extending conidiogenous cells.

Ellisembia henanensis is morphologically similar to E. ellipsoidea W.P. Wu

in conidial shape. However, E. ellipsoidea can be easily separated by its larger,

7-8-distoseptate conidia (40-45 x 10-11 um; Wu & Zhuang 2005).

Lylea tetracoila (Corda) Hol.-Jech., Folia Geobot. Phytotax. 13: 437 (1978). Fie. 2

CoLonizs on dead wood effuse, dark brown, hairy. Mycelium superficial

and immersed, septate, pale brown, smooth-walled hyphae, 1.5-2 um thick.

Ellisembia henanensis sp. nov. (China) ... 599

Fic. 1. Ellisembia henanensis (ex holotype, HSAUP H6697). A. Conidiophores, conidiogenous

cells, and conidia; B. conidiophores and conidiogenous cells; C. conidia.

600 ... Xia & al.

Fic. 2. Lylea tetracoila (ex HSAUP H6695). A. Conidiophores, conidiogenous cells, and

conidia; B. conidiophores and conidiogenous cells; C. conidia.

Ellisembia henanensis sp. nov. (China) ... 601

CONIDIOPHORES macronematous, mononematous, cylindrical, unbranched,

3-6-septate, smooth, pale brown, 30-60 x 3-3.5 um. CONIDIOGENOUS CELLS

monoblastic, integrated, terminal, pale brown, 10-15 x 3-3.5 um. Conidial

secession schizolytic. Conrp1a holoblastic, solitary, dry, acrogenous, fusiform

to cylindrical, 3-4-distoseptate, smooth-walled, pale brown, 20-40 x 4-6 um.

SPECIMEN EXAMINED: CHINA, HENAN PROVINCE: Jigong Mountain, on dead stems of

unidentified broadleaf tree, 16 Oct. 2015, J.W. Xia (HSAUP H6695).

ComMENTS—Lylea was established by Morgan-Jones (1975) with L. catenulata

Morgan-Jones as the type species. The genus is characterized by the formation

of catenate, distoseptate conidia from monoblastic conidiogenous cells. Six

species have been described (Morgan-Jones 1975, Holubova-Jechova 1978,

Mercado-Sierra et al. 1997, McKenzie 2009, Karandikar & Singh 2010, Xia et

al. 2014).

Lylea tetracoila is reported for the first time from China. Our specimen

closely matches the description by Holubova-Jechova (1978), which reported

conidia measuring (17.5—)20-40(-65) x (3-)4-5.5(-7) um. We believe they are

conspecific.

Menispora glauca Pers., Mycol. Europ. 1: 32 (1822). FIG. 3

CoLonliEs effuse, cushion-like, velvety to cottony, easily separable from

the substratum, greyish to blue-grey when sporulating, dark grey-brown

when old. Hyphae branched, pale to brown, 2-3 um wide, septate, superficial

and immersed in the substrate. CONIDIOPHORES setiform, erect, mostly

unbranched, sometimes branched in the lower parts, whip-like twisted toward

the apex, dark brown, septate, smooth, 270-450 x 3-4 um, rounded apex up

to 2-3 um wide. CONIDIOGENOUS CELLS monophialidic, intercalary, discrete,

determinate, pale brown, 15-25 x 3-4 um. Conrpia hyaline, cylindrical-

allantoid, slightly tapered and rounded ends, curved, 3-septate, 15-20 x 3.5-4.5

um, with a single, straight or gently curved setula, 4—-8.5 tm long at each end.

SPECIMEN EXAMINED: CHINA, HENAN PROVINCE: Jigong Mountain, on dead stems of

an unidentified broadleaf tree, 16 Oct. 2015, J.W. Xia (HSAUP H6699).

ComMENTS—The genus Menispora, initially erected by Persoon (1822),

was re-described and revised by Hughes & Kendrick (1968) and Holubova-

Jechova (1974) to include the following characteristics. Setae, which are either

sterile independent structures or hair-like sterile extensions of the setiform

conidiophores, are straight (slightly bent to sinuous) or variously coiled and

twisted. Conidiogenous cells are terminal and integrated or intercalary and

discrete on branched conidiophores. When the conidiophore is setiform, the

phialidic conidiogenous cells are (i) lateral and sessile, or (ii) terminal on short

602 ... Xia & al.

20 um

Fic. 3. Menispora glauca (ex HSAUP H6699). A. Conidiophore (distal end of sterile extension);

B. partial conidiophores, conidiogenous cells, and conidia; C. conidia.

Ellisembia henanensis sp. nov. (China) ... 603

simple stalks, or (iii) lateral and terminal on once- or twice-branched divergent

or crowded laterals. The conidiogenous cells are usually slightly or strongly

recurved at the apex.

Menispora glauca is reported for the first time from China. Our specimen fits

well with the description by Holubova-Jechova (1974), which reported conidia

measuring 17—24(-27) x (3.7—)4-5 um. We conclude that they are conspecific.

Acknowledgments

The authors express gratitude to Dr. De-Wei Li and Dr. Rafael F Castafeda-Ruiz for

serving as pre-submission reviewers and for their valuable comments and suggestions.

This project was supported by the National Natural Science Foundation of China

(Nos. 31093440, 31230001, 31493010, 31493011) and the Ministry of Science and

Technology of the People’s Republic of China (No. 2006FY 120100).

Literature cited

Holubova-Jechova V. 1974. Lignicolous hyphomycetes from Czechoslovakia 4. Menispora. Folia

Geobotanica et Phytotaxonomica 8: 317-336. http://dx.doi.org/10.1007/BF02852830

Holubova-Jechova V. 1978. Lignicolous hyphomycetes from Czechoslovakia 5. Septonema,

Hormiactella, and Lylea. Folia Geobotanica et Phytotaxonomica 13: 421-442.

http://dx.doi.org/10.1007/BF02851944

Hughes SJ, Kendrick WB. 1968. New Zealand fungi. 12. Menispora, Codinaea, Menisporopsis. New

Zealand Journal of Botany 6: 323-375. http://dx.doi.org/10.1080/0028825X.1968.10428818

Karandikar KG, Singh SK. 2010. Lylea indica: a new hyphomycetes species from India. Mycotaxon

112: 257-260. http://dx.doi.org/10.5248/112.257

McKenzie EHC. 2009. A new species of Lylea (hyphomycetes) on Rhopalostylis (Arecaceae) in New

Zealand. Mycotaxon 109: 39-42. http://dx.doi.org/10.5248/109.39

Mercado-Sierra A, Figueras MJ, Gené J. 1997. New or rare hyphomycetes from Cuba VIII. Species

of Lylea, Phaeoisaria, Arxiella, Graphium, Periconia and Ramichloridium. Mycotaxon 63:

369-375.

Morgan-Jones G. 1975. Notes on hyphomycetes. VIII. Lylea, a new genus. Mycotaxon 3: 129-132.

Persoon CH. 1822. Mycologia Europaea 1. Erlangen. 356 p.

Shoemaker RA, Hambleton S. 2001. “Helminthosporium” asterinum, Polydesmus elegans, Imimyces,

and allies. Canadian Journal of Botany 79: 592-599. http://dx.doi.org/10.1139/b01-038

Subramanian CV. 1992. A reassessment of Sporidesmium (hyphomycetes) and some related taxa.

Proceeding of the Indian National Science Academy, B 58: 179-190.

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

Diversity Research Series 15. 351 p.

Xia JW, Ma YR, Zhang XG. 2014. New species of Corynesporopsis and Lylea from China. Sydowia

66(2): 241-248.

MY COTAXON

July-September 2016— Volume 131, pp. 605-611

http://dx.doi.org/10.5248/131.605

Thozetella coronata and T. ypsiloidea spp. nov.

from the Brazilian Amazon forest

JOSIANE SANTANA MONTEIRO’, RAFAEL F. CASTANEDA-RUIZ?

& Luis FERNANDO PASCHOLATI GusMAO"

' Universidade Estadual de Feira de Santana,

Avenida Transnordestina, s/n, Novo Horizonte, 44036-900, Feira de Santana, Brazil

? Instituto de Investigaciones Fundamentales en Agricultura Alejandro de Humboldt’ (INIFAT),

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

“ CORRESPONDENCE TO: gusmao@uefs. br

ABSTRACT— Two new species, Thozetella coronata and T. ypsiloidea collected from submerged

leaf litter in the Brazilian Amazon forest, are described and illustrated. Thozetella coronata is

characterized by the production of clavate coronate hyaline microawns. Thozetella ypsiloidea

is distinguished by finely verrucose Y-shaped hyaline microawns.

KEY worDs—Ascomycota, Chaetosphaeriaceae, submerged litter, taxonomy, tropical fungi

Introduction

The Amazon forest is regarded as one of the most important reservoirs of

biodiversity in the world (Peres et al. 2010). During a survey of microfungi

associated with submerged decaying plant materials from the Brazilian

Amazon, several unusual Thozetella specimens were found, two of which

represent new species and are here described and illustrated.

Thozetella Kuntze is distinguished by the presence of sporodochial,

synnematal, or effuse conidiomata, phialidic conidiogenous cells that

produce unicellular setulate lunate hyaline conidia, and sterile elements called

microawns. Microawns are a diagnostic feature of Thozetella (Seifert et al. 2011),

and 19 species have been accepted in this genus (Index Fungorum 2016), with

microawn morphology (Fic. 1) being the primary basis of species delimitation

(Silva & Grandi 2013).

606 ... Monteiro, Castahteda-Ruiz & Gusmio

ee ee re

Thozetella spp. nov. (Brazil) ... 607

Materials & methods

Samples of submerged decaying plant materials were collecting from streams in

the “Area de Protecao Ambiental Ilha do Combu’, Para State, Brazil. Samples were

treating according to Castaneda Ruiz et al. (2016) and examined periodically under

the stereomicroscope for 30 days. Semi-permanent and permanent microscope slides

of fungi were mounted in 100% lactic acid and PVL resin (alcohol polyvinyl, lactic

acid, and phenol). The specimens were identified by examination under a compound

microscope and microscopic elements were measured at a magnification of x1000.

Photomicrographs were obtained with a Olympus BX51 microscope equipped with

bright-field and Nomarski interference optics and a microscopy image acquisition DP25

digital color camera. The specimens are deposited in the Herbarium of Universidade

Estadual de Feira de Santana, Bahia, Brazil (HUEFS).

Taxonomy

Thozetella coronata J.S. Monteiro, R.F. Castafieda & Gusmao, sp. nov. Fia. 2

MycoBank MB 814790

Differs from all other Thozetella species by its clavate, coronate microawn.

Type: Brazil, Para State: Area de Protecdo Ambiental Ilha do Combu, 1°29’S 48°25’W,

on unidentified submerged leaves, 17.1.2011; coll. J.S. Monteiro (Holotype: HUEFS

196471).

EryMo_oey: Latin, coronata, in reference to the crown-like apical projections on the

microawns.

CoLonizs on the natural substrate cream-coloured. Mycelium immersed;

hyphae septate, branched, cylindrical cells, 1.5-2.5 um diam., smooth-walled,

pale brown. Conip1oMatTA sporodochial, superficial, scattered, with a white

conidial mass at the apex, 62.5-75 x 175-200 um diam. CONIDIOPHORES

macronematous, septate, cylindrical, smooth, pale brown. CONIDIOGENOUS

CELLS monophialidic, integrated, determinate, terminal, cylindrical, smooth,

pale brown, 13-15 x 2-3 um, lacking collarette. Conrp1 lunate, aseptate,

smooth, hyaline, 16-20 x 2-3 um, with a single filiform setula at each end,

6-8 um long. MicRoawns mixed and immersed in the conidial mass on the

natural substrate, aseptate, clavate, smooth, hyaline, 10-15 x 2-3 um; apical

part swollen, 4-6 um wide, with 5-6 coronate projections, 1-1.5 um long; base

1-1.5 um wide. Teleomorph unknown.

Fic 1. Microawns of Thozetella spp.: A. T. acerosa (Paulus et al. 2004). B. T. aculeata (Silva &

Grandi 2011). C. T. boonjiensis (Paulus et al. 2004). D. T. buxifolia (Allegrucci et al. 2004).

E. T. canadensis (Nag Raj 1976). E T. capitata (Silva & Grandi 2013). G. T. coronata (this work).

H. T. cristata (Pirozynski & Hodges 1973). I. T: cubensis (Castafteda & Arnold 1985). J. T. effusa (Sutton

& Cole 1983). K. T. falcata (Paulus et al. 2004). L. T: gigantea (Paulus et al. 2004). M. T. havanensis

(Castaneda 1984). N. T. nivea (Pirozynski & Hodges 1973). O. T. pinicola (Jeewon et al. 2009).

P. T. queenslandica (Paulus et al. 2004). Q. T. radicata (Morris 1956, as Neottiosporella radicata).

R. T. serrata (Whitton et al. 2012). S. T’ submersa (Barbosa et al. 2011). T. T: tocklaiensis (Agnihothrudu

1958, as Thozetellopsis tocklaiensis).U. T: ypsiloidea (this work). Scale bars = 20 um.

608 ... Monteiro, Castafeda-Ruiz & Gusmao

Fic. 2. Thozetella coronata (holotype: HUEFS 196471). a. Sporodochial conidioma.

b. Conidiogenous cells. c. Conidia with setulae. d. General aspect of microawns. e. Detail of

microawns. f—j. Microawns. Scale bars: a = 50 um; b-j = 10 um.

NotE: All previously described Thozetella species differ from T. coronata by the

absence of clavate, coronate microawns (Barbosa et al. 2011, Silva & Grandi

2013).

Thozetella ypsiloidea J.S. Monteiro, R.F. Castafieda & Gusmao, sp. nov. Fic. 3

MycoBAnk MB 814791

Differs from all other Thozetella species by its Y-shaped microawns.

Type: Brazil, Para State: Area de Protecao Ambiental Ilha do Combu, 1°29’S 48°25’W,

on unidentified submerged leaves, 18.X.2011; coll. J.S. Monteiro (Holotype: HUEFS

196464).

EryMo.ocy: Latin, ypsiloidea, in reference to the Y-shaped microawns.

Thozetella spp. nov. (Brazil) ... 609

Fic. 3. Thozetella ypsiloidea (holotype: HUEFS 196464). a. Sporodochial conidioma. b. General

aspect of conidiophores. c. Conidia with setulae. d-i. Microawns. Scale bars: a = 50 um; b = 20 um;

c-i= 10 um.

CoLonigs on the natural substrate cream-coloured. Mycelium immersed;

hyphae septate, branched, cylindrical cells, 1.5-2 um diam., smooth-walled,

pale brown. ConipiomatTa sporodochial, superficial, scattered with a white

conidial mass at the apex, 42.5-87.5 x 47.5-150 um diam. CONIDIOPHORES

macronematous, septate, cylindrical, smooth, pale brown. CONIDIOGENOUS

CELLS monophialidic, integrated, determinate, terminal, cylindrical, smooth,

pale brown, 12-25 x 2-3 um, lacking collarette. Conrp1A lunate, aseptate,

smooth, hyaline, 11-15 x 2-3 um, with a single filiform setula at each end, 5-8

uum long. MicRoawns mixed and immersed in mass on the natural substrate,

610 ... Monteiro, Castafeda-Ruiz & Gusmao

aseptate, Y-shaped, finely verrucose on arm which has rounded apex and

partially verrucose or smooth on the other arm, partially smooth on stipe,

hyaline, 12-22 x 8-13 um; 2-4 um wide at the base. Teleomorph unknown.

Note: All previously described Thozetella species differ from T. ypsiloidea by

the absence of Y-shaped microawns (Barbosa et al. 2011, Silva & Grandi 2013).

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 thank to the National Council

for Scientific and Technological Development (CNPq) (Proc. 142014/2011-7) for

supporting this study. The first author is grateful to Coordination for the Improvement

of Higher Education Personnel (CAPES) for the scholarship. 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.

We acknowledge the facilities provided by Dr. P.M. Kirk and Dr. V. Robert, through

the Index Fungorum and MycoBank websites. Dr. Lorelei Norvell’s editorial review and

Dr. Shaun Pennycook’s nomenclature review are greatly appreciated.

Literature cited

Agnihothrudu V. 1958. Notes on fungi from north-east India. I. A new genus of Tuberculariaceae.

Mycologia 50(4): 570-579. http://dx.doi.org/10.2307/3756119

Allegrucci N, Cazau MC, Cabello MN, Arambarri A. 2004. Thozetella buxifolia sp. nov., a new

hyphomycete from Argentina. Mycotaxon 90(2): 275-279.

Barbosa FR, Silva SS, Fiuza PO, Gusmao LFP. 2011. Conidial fungi from the semi-arid

Caatinga biome of Brazil. New species and records for Thozetella. Mycotaxon 115: 327-334.

http://dx.doi.org/10.5248/115.327

Castaneda Ruiz RE. 1984. Nuevos taxones de Deuteromycotina: Arnoldiella robusta gen. et sp. nov.;

Roigiella lignicola gen. et sp. nov.; Sporidesmium pseudolmediae sp. nov. y Thozetella havanensis

sp. nov. Revista del Jardin Botanico Nacional, Universidad de La Habana 5(1): 57-87.

Castaneda Ruiz RF, Arnold GRW. 1985. Deuteromycotina de Cuba. I. Hyphomycetes. Revista del

Jardin Botanico Nacional, Universidad de La Habana 6(1): 47-67.

Castafieda Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and South

America. 197-217, in: DW Li (ed.). Biology of Microfungi, Springer International Publishing.

http://dx.doi.org/10.1007/978-3-319-29137-6_9

Index Fungorum. 2016. http://www.indexfungorum.org/Names/Names.asp (accessed 22 Apr.

2016).

Jeewon R, Yeung SYQ, Hyde KD. 2009. A novel phylogenetic group within Thozetella

(Chaetosphaeriaceae): a new taxon based on morphology and DNA sequence analyses.

Canadian Journal of Microbiology 55(6): 680-687. http://dx.doi.org/10.1139/W08-148

Morris EF. 1956. Tropical fungi imperfecti. Mycologia 48(5): 728-737.

http://dx.doi.org/10.2307/3755382

Nag Raj TR. 1976. Miscellaneous microfungi. I. Canadian Journal of Botany 54(12): 1370-1376.

http://dx.doi.org/ 10.1139/b76-149

Paulus B, Gadek P, Hyde KD. 2004. Phylogenetic and morphological assessment of five

new species of Thozetella from an Australian rainforest. Mycologia 96(5): 1074-1087.

http://dx.doi.org/10.2307/3762090

Thozetella spp. nov. (Brazil) ... 611

Peres, CA, Gardner TA, Barlow J, Zuanon J, Michalski F, Lees AC, Vieira ICG, Moreira FMS,

Feeley KJ. 2010. Biodiversity conservation in human-modified Amazonian forest landscapes.

Biological Conservation 143: 2314-2327. http://dx.doi.org/10.1016/j.biocon.2010.01.021

Pirozynski KA, Hodges CS Jr. 1973. New hyphomycetes from South Carolina. Canadian Journal of

Botany 51(1): 151-173. http://dx.doi.org/10.1139/b73-024

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

Biodiversity Series 9. 997 p.

Silva P, Grandi RAP. 2011. A new species of Thozetella from Brazil. Cryptogamie Mycologie 32:

359-363. http://dx.doi.org/10.7872/crym.v32.iss4.2011.359

Silva P, Grandi RAP. 2013. Taxonomic studies of Thozetella Kuntze (anamorphic Chaetosphaeriaceae,

Ascomycota). Nova Hedwigia 97(3-4): 361-399.

http://dx.doi.org/10.1127/0029-5035/2013/0121

Sutton BC, Cole GT. 1983. Thozetella (Hyphomycetes): an exercise in diversity. Transactions of the

British Mycological Society 81(1): 97-107. http://dx.doi.org/10.1016/S0007-1536(83)80209-5

Whitton SR, McKenzie EHC, Hyde KD. 2012. Anamorphic fungi associated with Pandanaceae.

Fungal Diversity Research Series 21: 125-353. http://dx.doi.org/10.1007/978-94-007-4447-9_4

MY COTAXON

July-September 2016— Volume 131, pp. 613-619

http://dx.doi.org/10.5248/131.613

Long-hidden in Thaxter’s treasure trove:

Laboulbenia camerunensis sp. nov.

parasitic on African Curculionidae

TRISTAN W. WANG!, DANNY HAELEWATERS”* & DONALD H. PFISTER?

"Harvard College, 365 Kirkland Mailing Center, Cambridge, Massachusetts 02138, U.S.A.

? Farlow Reference Library and Herbarium of Cryptogamic Botany, Harvard University,

22 Divinity Avenue, Cambridge, Massachusetts 02138, U.S.A.

" CORRESPONDENCE TO: dhaelewaters@fas.harvard.edu

ABSTRACT—A new species, Laboulbenia camerunensis, parasitic on Curculio sp. from

Cameroon, is described from a historical slide prepared by Roland Thaxter. It is the seventh

species to be described from the family Curculionidae worldwide and the first from the

African continent. The species is recognized by the characteristic outer appendage. The

latter consists of two superimposed hyaline cells, separated by a black constricted septum,

the suprabasal cell giving rise to two branches, the inner of which is simple and hyaline, and

the outer tinged with brown. A second blackish constricted septum is found at the base of

this outermost branch. Description, illustrations, and comparison to other species are given.

Key worps—Laboulbeniales, Laboulbeniomycetes, insect-associated fungi, taxonomy, weevils

Introduction

The order Laboulbeniales (Fungi, Ascomycota, Laboulbeniomycetes) consists

of microscopic ectoparasites that develop on the exoskeleton of certain

invertebrates. The hosts are primarily beetles but also include millipedes,

mites, and a variety of insects (flies, ants, cockroaches, and others). Unlike

other multicellular fungi, Laboulbeniales exhibit determinate growth, meaning

that the fungal body (thallus) develops from a two-celled ascospore through a

restricted and regulated number of mitotic divisions to produce an individual

with a set number of distinctively arranged cells (Tavares 1985, Santamaria

1998).

614 ... Wang, Haelewaters & Pfister

Although the earliest observations on these parasites occurred in the 1840s

(Rouget 1850), it was not until the research of Roland Thaxter [1858-1932]

that the Laboulbeniales were studied in depth. In 1890 Thaxter published the

first in a series of twenty-one non-illustrated papers in which hundreds of new

species were described. His five-volume illustrated monograph, published

between 1896 and 1931, forms the basis for all later studies of the group. Since

Thaxter died one year after the publication of the fifth volume, a planned sixth

volume was never prepared. This was to have been a synthesis of the large genus

Laboulbenia and “a general review, classification, and host-index” (Thaxter

1931). Although Thaxter left no manuscript for this final volume (Benjamin

1971), the Farlow Reference Library of Cryptogamic Botany archived his

sketches and notes on undescribed species, many of which were recently

re-examined along with his slide preparations.

Our recent interest in Thaxter’s permanent slide collection has resulted in

the description of several new species of Laboulbeniales on beetle hosts in the

families Carabidae (Laboulbenia poplitea Haelew.), Chrysomelidae (Laboulbenia

bilobata Haelew. & W. Rossi, L. longipilis Haelew. & W. Rossi, L. pfisteri

Haelew. & W. Rossi), Erotylidae (Laboulbenia erotylaridarum Haelew.), and

Hydrophilidae (Zodiomyces rhizophorus W. Rossi et al.) (Haelewaters & Yaakop

2014, Haelewaters & Rossi 2015, Rossi et al. 2016b). Further examination of

the slides revealed yet another undescribed species of Laboulbenia, on Curculio

sp., a member of the family Curculionidae (Wang 2014). Although reported

by his contemporary Spegazzini (1917) from Argentina and more recently by

Sugiyama & Majewski (1987) and Rossi et al. (2015), this is the only material

studied by Thaxter that occurs on a member of the Curculionidae.

Currently about 2100 species of Laboulbeniales have been described.

Species descriptions in Laboulbeniales are based exclusively on morphological

characters, except for two species (Hesperomyces coleomegillae W. Rossi &

A. Weir, H. palustris W. Rossi & A. Weir) whose morphological descriptions

are supported by molecular characters (Goldmann et al. 2013). Generating

sequences of Laboulbeniales has not been easy, although recent endeavors

offer hope for advancing molecular phylogenetic research in the group when

adequate fresh material is available (Haelewaters et al. 2015).

Although many authors agree that adequate species descriptions require

the study of multiple juvenile and mature thalli, quite a few descriptions have

been based on only one, two, or very few thalli. In these cases, morphology

should be unusual and distinctive enough to make a strong case for describing

the species. Examples where limited material has been used for description

are Cantharomyces magellanicus Thaxt. (3 thalli studied, of which only one

was mature, but damaged), Dimeromyces osellae W. Rossi (1 female and

Laboulbenia camerunensis sp. nov. (Cameroon) ... 615

1 male thallus), Laboulbenia arietina Thaxt. (4 thalli), L. longipilis (2 thalli),

Monoicomyces denticulatus Thaxt. (3 thalli, of which two were mature), and

Peyritschiella xyricola Thaxt. (1 thallus) (Haelewaters & Rossi 2015; Rossi 2010;

Thaxter 1914, 1915, 1931). A more extreme case is Peyritschiella lampropygi

Thaxt., which was described on the basis of material that was not fully developed

(Thaxter 1931). For the description of the genus Columnomyces R.K. Benj., only

a single mature (and broken) thallus was available among the 19 immature/

broken thalli (Benjamin 1955).

One difficulty with our new species is that we did not have multiple adult

specimens, but the available material does include one mature thallus in

addition to several immature thalli. Nonetheless, because this species is easily

distinguished from other Laboulbenia species based on the characters of the

outer appendage, we describe it here as a new species. Such records help to

direct collecting and ultimately inform diversity studies.

Materials & methods

Material was discovered amongst unidentified mounts studied by Roland Thaxter in

the permanent slide collection, deposited at the Farlow Herbarium, Harvard University

(FH).

Observations and measurements were made using an Olympus BX40 light

microscope with Olympus XC50 digital camera and MicroSuite Special Edition software

3.1 (Soft Imaging Solutions GmbH). Line drawings were made using a drawing tube.

Illustrations were optimized (using LEVELS and BRIGHTNESS/CONTRAST tools), cropped,

and edited in Adobe Photoshop CS Version 8.0 (San Jose, California).

Taxonomy

Laboulbenia camerunensis T.W. Wang & Haelew., sp. nov. PLATE 1

MycoBAnk 811237

Differs from all other Laboulbenia species on Curculionidae by its striking outer

appendage.

Type: Cameroon, on the abdomen of Curculio sp. (family Curculionidae, subfamily

Curculioninae), Jun. 1912, no collector, Thaxter 2360 (Holotype, FH 00313482 [slide,

in poor condition, partly dried out; 1 mature thallus and 10 thalli in various stages of

development]).

Erymo ocy: The epithet camerunensis is derived from the country where the holotype

was collected.

THALLUs hyaline, with the exception of the dark brown outermost branch of

the outer appendage; 160 um long from foot to perithecial tip. CeLis I and II of

similar length, up to 3x as long as broad, with subparallel margins; forming an

elongated pedestal. CExt III slightly longer than broad, distally widening. CELL

IV about as long as cell II or somewhat shorter. CELL V wedge-shaped, almost

616 ... Wang, Haelewaters & Pfister

as long as cell IV; septum IV-V is oblique and reaches cell III. INSERTION CELL

black, flattened, not marking an evident constriction on the posterior margin of

the thallus. OUTER APPENDAGE consisting of two superimposed almost hyaline

cells separated by a black constricted septum; two simple branches arising from

the suprabasal cell, composed of cells much longer than broad; the septum

separating the suprabasal cell and the lower cell of the outermost branch, black,

often oblique, and the area above tinged with dark brown. INNER APPENDAGE

with very small basal cell, giving rise to 2-3 small cells, longer than broad,

each carrying one flask-shaped antheridium, separated by a thin, dark-brown

septum. CELL VI quadrangular with subequal width and length. PERITHECIUM

fusiform, largely free, 65 x 30 um; strongly tapered at the tip, ending in four

hyaline and rounded lips, two of which carry small papillae; preostiolar spots

reduced to only the posterior one, which consists of a shaded area occupying

the posterior lip.

Discussion

We think it is safe to assume that the host genus is correctly identified as

Curculio Linnaeus 1758, according to current taxonomical concepts. The

superfamily Curculionoidea (Coleoptera) comprises over 60,000 species

of weevils, most of which represent the Curculionidae (true weevils). The

phylogenetic classification of this family is complex, leading to recent debate

(Jordal et al. 2014). However, the taxonomy of the Curculionini tribe in Africa

has remained almost unchanged since Thaxter’s work (Lobl & Smetana 2013,

Caldara et al. 2014).

Six Laboulbenia species were previously described from Curculionidae:

L. curculionidicola K. Sugiy. & T. Majewski from Peru; L. dichroma W. Rossi et

al. on Geraeus sp. from Ecuador; L. elephantina W. Rossi et al. on Udeus sp. from

Ecuador; L. hyemalis Speg. on Anthonomus sp. from Argentina; L. inopinata

W. Rossi et al. on Lechriops spp. and Macrocopturus spp. from Ecuador; and

L. microcarpa W. Rossi et al. on Sicoderus sp. and Lancearius esau from Ecuador

(Spegazzini 1917, Sugiyama & Majewski 1987, Rossi et al. 2015). Laboulbenia

camerunensis can be easily distinguished from these species by its characteristic

PLaTE 1. Laboulbenia camerunensis (holotype, FH 00313482). A. Juvenile thallus, with trichogyne

(tr), an external thin appendage-like outgrowth of the young perithecium through which

fertilization occurs. B. Mature thallus, with the outermost branch of the outer appendage broken

off above the suprabasal cell (soa). Scale bars = 50 um. Labeled are: the foot (f) with which the

thallus attaches to the host’s integument; cells I, II, III, IV, and V of the receptacle; the insertion cell

(ic); the basal cell (boa), and suprabasal cell (soa) of the outer appendage, as well as its outermost

branch (00a) [only in A]; the very small basal cell of the inner appendage (bia), which carries

2 [in B] or 3 [in A] cells each supporting a single antheridium; and cell VI, carrying the perithecium

(including basal cells).

Laboulbenia camerunensis sp. nov. (Cameroon) ... 617

618 ... Wang, Haelewaters & Pfister

outer appendage structure, in particular the two dark septa at the base and the

outermost branch tinged with dark brown above its lower cell.

Laboulbenia curculionidicola is additionally distinguished from L. camerunensis

by cell II being twice as long as cell I, cell III usually splitting into two,

cell V located in the upper inner corner of cell IV, and the outer appendage

comprising a simple branch. Laboulbenia hyemalis is further distinguished by

its outer appendage consisting of a single main axis that is blackened only at the

posterior margin and its perithecial tip distinctly curved toward the anterior

side. Laboulbenia dichroma, L. elephantina, and L. microcarpa, among other

differences, have a simple, unbranched outer appendage. The final species,

L. inopinata, differs greatly from all other species on Curculionidae in its

undivided androstichum (= cells III + IV + V) and very simple appendage

structure. The undivided androstichum is actually typical for species of

Laboulbenia on Chrysomelidae (Rossi et al. 2016a).

The outer appendage of L. camerunensis, highly distinctive in Laboulbenia,

superficially resembles L. pfisteri, which, however, differs in many other

characteristics: (i) cell V does not reach cell III, (ii) the outer appendage has

an additional third branch borne on the inner angle of the basal cell, and (iii)

the perithecium is noticeably enlarged in its upper portion (Haelewaters &

Rossi 2015).

Roland Thaxter’s original sketch of this species, obtained from the archives

of the Farlow Reference Library of Cryptogamic Botany, was published in

Wang (2014: 3).

Acknowledgments

We wish to thank Lisa Decesare, Head of Archives and Public Services at the Botany

Libraries, Harvard University, for her efforts to access Roland Thaxter’s unfinished

drawings in the archives of the Farlow Reference Library. We also thank Kanchi N.

Ghandhi for nomenclatural advice, Meredith Blackwell and Walter P. Pfliegler for

reviewing the manuscript, and Shaun R. Pennycook for additional edits and suggestions.

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Benjamin RK. 1971. Introduction and supplement to Roland Thaxter’s Contribution towards a

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Systematics (Phytophaga). Berlin, Walter De Gruyter.

Goldmann L, Weir A, Rossi W. 2013. Molecular analysis reveals two new dimorphic

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Brill.

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la Société Entomologique de France 8: 21-24.

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Mycological Progress 9(3): 407-415. http://dx.doi.org/10.1007/s11557-009-0650-0

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rainforest. Phytotaxa 231(2): 187-192. http://dx.doi.org/10.11646/phytotaxa.231.2.8

Rossi W, Bernardi M, Torres JA. 2016a. New species of Laboulbenia parasitic on leaf beetles.

Mycological Progress 15(1): 4 [on-line, 7 p.]. http://dx.doi.org/10.1007/s11557-015-1146-8

Rossi W, Haelewaters D, Pfister DH. 2016b. Fireworks under the microscope: a spectacular

new species of Zodiomyces from the Thaxter collection. Mycologia 108: 709-715.

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Santamaria S. 1998. Laboulbeniales. 1. Laboulbenia. Flora Mycologica Iberica 4. 186 p.

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Historia Natural de Buenos Aires 29: 445-688.

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Ascomycotina) II. Transactions of the Mycological Society of Japan 28: 121-136.

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MY COTAXON

July-September 2016—Volume 131, pp. 621-629

http://dx.doi.org/10.5248/131.621

Erysiphe acantholimonis sp. nov. on Acantholimon hedinii

J1a-GE Sona’, BIAo Xu’, SHAN-HE ZHANG’,

ZHEN-YU ZHAO’, SUNG-EUN CHO? & HYEON-DONG SHIN”

‘College of Life Sciences, Tarim University & Province-Ministry Joint Key

Laboratory of Protection and Utilization of Biological Resources in Tarim Basin,

Alar 843300, China

Division of Environmental Science and Ecological Engineering, Korea University,

Seoul 02841, Korea

“ CORRESPONDENCE TO: hdshin@korea.ac.kr

ABSTRACT—A powdery mildew on Acantholimon hedinii was collected in China,

examined, described, and proposed as a new species, Erysiphe acantholimonis. Recognition

of this new species is supported by phylogenetic analyses of the internal transcribed spacer

regions including 5.88 and 28S rDNA sequences from three E. acantholimonis specimens.

A comparison with and a key to the three species of Erysiphe on hosts belonging to the

family Plumbaginaceae are provided.

Key worps—Erysiphaceae, Erysiphe aurea, Erysiphe limonii, ITS, molecular phylogeny

Introduction

Powdery mildews (Erysiphaceae) are holobiotrophic fungi pathogenic on

plant leaves, stems, flowers, and fruits. These pathogens cause moderate to

serious damage in nearly 10,000 species of angiosperms (Amano 1986, Glawe

2008). Braun (1987) introduced a classification of the Erysiphaceae that split

the family into 18 genera, including Erysiphe, Microsphaera, and Uncinula. On

the basis of phylogenetic analyses, Braun & Takamatsu (2000) subsequently

reduced these three genera to synonymy under Erysiphe (sensu lato), which

they separated into three non-phylogenetic sections: E. sect. Erysiphe, E. sect.

Microsphaera, and E. sect. Uncinula. Erysiphe sect. Erysiphe is characterized by

having chasmothecia with mycelioid appendages.

622 ... Song & al.

Two species of Erysiphe sect. Erysiphe, E. aurea R.Y. Zheng & G.Q. Chen

and E. limonii L. Junell, have been described on plants within the family

Plumbaginaceae. In 2010, a powdery mildew occurring on Acantholimon

(Plumbaginaceae) was found in Xinjiang, China, which Song et al. (2015)

reported as the first record of Erysiphe limonii on A. hedinii from China.

However, a detailed morphological re-examination and molecular analyses

have revealed that powdery mildew on this host is clearly distinct from

E. limonii. Here, we propose the fungus collected on A. hedinii as a new

species, E. acantholimonis, and base our description on a morphological

characterization and DNA sequence comparison. In addition, we discuss

relationships of the new species to closely related powdery mildew fungi.

Materials & methods

Fungal structures were detached from the infected leaves and mounted in a few drops

of distilled water on a glass slide for light microscopy. Morphological characteristics of

the fungal structures were examined in bright field and differential interference contrast

(DIC) light microscopy, using an Olympus BX51 microscope (Olympus, Tokyo, Japan)

for measurements and a Zeiss AX10 microscope equipped with AxioCam MRc5 (Carl

Zeiss, Gottingen, Germany) for photographs. Elements were measured at 100x, 200x,

400x, and 1000x magnifications. Representative voucher specimens were deposited in

the Mycological Herbarium of Tarim University, Xinjiang, China (HMUT), and the

Korea University Herbarium, Seoul, Korea (KUS).

Genomic DNA was extracted from chasmothecia using the Chelex 100 method as

set forth in Walsh et al. (1991) and Hirata & Takamatsu (1996). The 28S ribosomal

DNA (rDNA) including domains D1 and D2 and the internal transcribed spacer

(ITS) regions including ITS1, 5.8S, and ITS2 were amplified by polymerase chain

reaction (PCR) using the primer pairs ITS1/ITS4 (White et al. 1990) for ITS, and PM3

(Takamatsu & Kano 2001) and TW14 (Moriet al. 2000) for 28S. The PCR products were

then sequenced directly (Takamatsu et al. 2009) by a commercial sequencing company

(Macrogen, Seoul, Korea) using the same primers used for PCR. The resulting sequences

were deposited in GenBank (Accession Nos. KU554532-KU554537). The sequences

were used in a BLASTn search against the NCBI nucleotide databases to confirm

the identity of the isolates. The sequences were aligned with published sequences of

E. sect. Erysiphe species retrieved from GenBank, using the online version of MAFFT

ver. 7.215 (Katoh et al. 2002). Neoerysiphe galeopsidis (DC.) U. Braun was used as an

outgroup. Maximum Likelihood (ML) analyses were performed using RAxML-HPC

BlackBox ver. 8.2.4 (Stamatakis 2006, Stamatakis et al. 2008) in the CIPRES cluster

server (http://www.phylo.org/) at the San Diego Supercomputer Center.

Fic. 1. Erysiphe acantholimonis (holotype, HMUT00821). A: hypha with appressoria (arrows);

B, C: conidiophores; D: conidia (arrow indicates a primary conidium); E: germinating

conidium; F: surface structure of a wrinkled conidium; G: chasmothecium with appendages;

H: chasmothecium containing several asci; I, J: asci containing 3-4 ascospores; K: ascospores.

623

Erysiphe acantholimonis sp. nov. (China) ...

624 ... Song & al.

Taxonomy

Erysiphe acantholimonis J.G. Song, B. Xu & H.D. Shin, sp. nov. FIG. 1

MycoBAnk MB 815918

Differs from Erysiphe limonii by its larger conidial length/width ratio, its smaller

2-4-spored asci, and its Acantholimon host.

Type: China, Xinjiang, Wugia county, 39°53’17”N 74°56’47’E, on Acantholimon hedinii

Ostenf. (Plumbaginaceae), 29 Aug. 2010, B. Xu (holotype, HMUT00821; isotype,

KUS-F29047; GenBank KU554537, KU554536).

ErymMo.ocy: The epithet is derived from the host genus, Acantholimon.

CoLoNIEs amphigenous, thinly effused or conspicuous, forming circular to

irregular white patches. HYPHAE straight or almost so, branched, hyaline,

septate, 4-8 um diam. HyPHAL APPRESSORIA well-developed, moderatelylobed

to multi-lobed, solitary or in opposite pairs, 4-9 um diam. CONIDIOPHORES

arising from the upper part of mother cells, 72-105 x 6.5-9.0 um, producing

conidia singly. FooT-cELLs mostly straight or almost so, cylindrical, 17-38

um long, relatively short, followed by 2-3 shorter cells. Conrpra hyaline,

cylindrical-oval to oblong, 30-50 x 11-15 um with a length/width ratio of

2.0-4.1, lacking fibrosin bodies, producing germ tubes in a perihilar position,

with an angular/rectangular wrinkling pattern on the surface. PRIMARY

CONIDIA apically rounded, basally subtruncate, generally smaller than the

secondary conidia. CHASMOTHECIA amphigenous, cauligenous, gregarious,

dark brown, spherical, 80-130 um diam, containing 5-10 asci. PERIDIUM CELLS

irregularly polygonal or rounded, 10-20 um diam, inner wall of the peridium

cells yellowish to hyaline. APPENDAGES few to numerous, 0-1-septate,

mycelioid, 0.5-1 times as long as the chasmothecial diameter, 50-100 x

7.5-9.0 um, tips distinctly recurved, brown at the base, becoming paler toward

the tip. Asc1 obovoid to ellipsoid or saccate, mostly short-stalked, 52-68 x

32-40 um, containing 2-4 spores per ascus. Ascospores oblong-ovoid,

hyaline, guttulate, 25-32 x 12-15 um.

ADDITIONAL SPECIMENS EXAMINED: on Acantholimon hedinii: CHINA, XINJIANG,

Wugia county, 19 Jun. 2010, B. Xu (HMUT00507, KUS-F29044); 28 Aug. 2010, B. Xu

(HMUT01489, KUS-F29046; GenBank KF318724, KU554532); Wensu county, 24 Aug.

2010, B. Xu (HMUT00396, KUS-F29045; GenBank KU554535, KU554534).

HOST RANGE AND DISTRIBUTION: on Acantholimon hedinii (China, endemic).

Phylogenetic analyses

ADDITIONAL SPECIMEN SEQUENCED: Erysiphe limonii: KOREA, IKsAn, 11 May 2013,

H.D. Shin (KUS-F27301; GenBank KU554533).

The GenBank BLAST search of the examined powdery mildew isolates showed

that E. acantholimonis on A. hedinii belongs to E. sect. Erysiphe. Based on the

Erysiphe acantholimonis sp. nov. (China) ... 625

too | JX235964 Erysiphe magnoliae

JX235965 E. magnoliae

57 AF049331 E. necator

AB015934 E. glycines

LC009939 E. juglandis

97) EU 140958 E. cruciferarum

EF592611 E. cruciferarum

eat 10 LC009959 E. buhrii

%8| 'L.C009960 E. buhrii

LC010013 E. polygoni

‘ KF318724 E. limonii ex Limonium latifolium x bellidifolium

81} -4LC010038 E. limonii ex Limonium sp.

LC010039 E. limonii ex L. platyphyllum

AB331648 E. nomurae

100 |AB257437 E. paeoniae

AB257436 E. paeoniae

100) AY870864 E. pulchra

LC009925 E. pulchra

AB498945 Neoerysiphe. galeopsidis

0.06

Fic. 2. ITS (including 5.88 rDNA) phylogenetic relationship between Erysiphe acantholimonis

isolates and reference isolates retrieved from GenBank, inferred by RAXML analysis. Sequences

of E. acantholimonis are shown in bold and their clade is highlighted in gray. Numbers above the

branches represent bootstrap support values >50% obtained from 1000 bootstrap replicates. Scale

bar indicates the number of substitutions per site.

molecular phylogenetic trees generated from the RAxML analyses of the ITS

and 28S rDNA, sequences generated from the three E. acantholimonis isolates

formed an independent clade with bootstrap (BS) supports of 99% for ITS (Fic.

2) and 86% for 28S (Fic. 3). The clade of the new fungus was clearly separated

from E. limonii from Korea (KF318724 and KU554533 ex Limonium latifolium

x bellidifolium) and Ukraine (LC010038 ex Limonium sp.; and LC010039 ex

L. platyphyllum) in the RAxML trees. In addition, E. acantholimonis was shown

to be a sister species to E. limonii with BS supports of 95% for ITS (Fic. 1) and

98% for 28S (Fic. 3), suggesting a close phylogenetic relationship. Thus the

status of E. acantholimonis as a new species is fully supported by the molecular

sequence analyses.

626 ... Song & al.

97| LC009985 Erysiphe cruciferarum

AB102944 iFe

LC010039 E. limonii ex Limonium platyphyllum

KU554533-E. limonii ex L. latifolium x bellidifolium

LC010038 E. limonii ex Limonium sp.

LC009959 E. buhtii

AB022382 E. friesii var. dahurica

AB257438 E. paeoniae

LC009910 E. glycines

LC009939 E. juglandis

98) AB571053.E. ligusiri

AB571057 E. ligustri

JX235969 E. magnoliae

JX235965 E, magnoliae

AB498950. Neoerysiphe galeopsidis

0.04

Fic. 3. 28S rDNA phylogenetic relationships between Erysiphe acantholimonis isolates and reference

isolates retrieved from GenBank, inferred by RAxML analysis. Sequences of E. acantholimonis

are shown in bold and their clade is highlighted in gray. Numbers above the branches represent

bootstrap support values >50% obtained from 1000 bootstrap replicates. Scale bar indicates the

number of substitutions per site.

Discussion

Two Erysiphe species belonging to E. sect. Erysiphe have previously been

recorded as occurring on Plumbaginaceae worldwide (Braun & Cook 2012,

Farr & Rossman 2016). Erysiphe limonii, found from Europe to Asia on various

Limonium spp., is morphologically similar to E. aurea but differs by having

appendages with some septa, 3-8 asci, and an inner peridial wall that is not

golden yellow (Braun & Cook 2012). The phylogenetic status of E. limonii

was recently resolved (Takamatsu et al. 2015, Choi et al. 2016), resulting in

a characterization based on morphological observations and a molecular

sequence analysis. Erysiphe aurea was described from a single collection

on Limonium suffruticosum in China (Zheng & Chen 1980, Braun & Cook

2012); it is distinguished from E. limonii in having aseptate appendages and

an inner peridial wall composed of golden-yellow cells. The conidial state of

E. aurea is inadequately known, and molecular data are not yet available for

Erysiphe acantholimonis sp. nov. (China) ... 627

phylogenetic analyses. Therefore, additional material is needed to ascertain the

taxonomic status of E. aurea, and to clarify its relationship to E. limonii and

E. acantholimonis.

Erysiphe limonii differs from E. acantholimonis by its larger, 3-6-spored

asci (45-85 x 25-50 um) and smaller conidial length/width ratio (1.6-2.9;

Choi et al. 2016), while E. aurea differs from E. acantholimonis by its longer

and narrower (60-80 x 25-35 um) 4—-6-spored asci and golden yellow inner

peridial wall.

Acantholimon comprises about 200 species worldwide (Dogan & Akaydin

2004). Recent molecular phylogenetic studies (Lledo et al. 2005, Moharrek et

al. 2014) have shown that Limonium and Acantholimon are not closely related,

suggesting that the Acantholimon and Limonium powdery mildews are unlikely

to be accepted as conspecific. Amano (1986) listed Acantholimon avenaceum,

A. kotschyi, and Acantholimon sp.as hosts of E. limonii. Additional morphological

and molecular data are needed to supplement our current knowledge about

powdery mildews infecting Acantholimon spp.

DNA sequence comparisons based on the ITS and 28S rDNA data obtained

from the analyzed Acantholimon specimens place their powdery mildew

fungus in E. sect. Erysiphe. In addition, the results indicate that E. limonii

and E. acantholimonis may have diverged along with their plumbaginaceous

hosts. Results obtained from the phylogenetic analyses are further supported

by morphological data, showing that E. acantholimonis is a distinct species

occurring on A. hedinii in China. Based on the information obtained from

literature as well as our own data, we provide a key to the three species of

Erysiphe occurring on Plumbaginaceae:

Key to Erysiphe species known on Plumbaginaceae

1. Inner peridial wall golden yellow; appendages aseptate .................. E. aurea

1. Inner peridial wall not golden yellow; appendages at least partly septate ......... 2

2. Asci relatively large (45-85 x 25-50 um), 3-6-spored;

median conidial length/width ratio <3.0 ............. 0... eee eee E. limonii

2. Asci relatively small (52-68 x 32-40 um), 2-4-spored;

median conidial length/width ratio mostly >3.0 ............ E. acantholimonis

Acknowledgements

The authors are grateful to Uwe Braun (Martin Luther University, Halle, Germany)

and Susumu Takamatsu (Mie University, Tsu, Japan) for critical comments on the

manuscript. This work was supported by the National Natural Science Foundation of

China (Nos. 31360007, 31460008) and the BK21 Plus program in 2016-2020 funded by

the National Research Foundation of Korea (NRF).

628 ... Song & al.

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700 p.

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildews). CBS

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Braun U, Takamatsu S. 2000. Phylogeny of Erysiphe, Microsphaera, Uncinula (Erysipheae) and

Cystotheca, Podosphaera, Sphaerotheca (Cystotheceae) inferred from rDNA ITS sequences

— some taxonomic consequences. Schlechtendalia 4: 1-33.

Choi IY, Choi CH, Cho SE, Park JH, Shin HD. 2016. First report of powdery mildew

caused by Erysiphe limonii on perennial hybrid statice in Korea. Plant Dis. 100: 652.

http://dx.doi.org/10.1094/PDIS-08-15-0893-PDN

Dogan M, Akaydin G. 2004. Three new species with two flowered spikelets in Acantholimon

(Plumbaginaceae) from East Anatolia, Turkey. Bot. J. Linn. Soc. 144: 497-505.

http://dx.doi.org/10.1111/j.1095-8339.2003.00267.x

Farr DE, Rossman AY. 2016. Fungal databases. Syst. Mycol. Microbiol. Lab., Online publication,

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Glawe DA. 2008. The powdery mildews: a review of the world’s most familiar (yet poorly

known) plant pathogens. Ann. Rev. Phytopathol. 46: 27-51.

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Hirata T, Takamatsu S. 1996. Nucleotide diversity of rDNA internal transcribed spacers extracted

from conidia and cleistothecia of several powdery mildew fungi. Mycoscience 37: 283-288.

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alignment based on fast Fourier transform. Nucleic Acids Res. 30: 3059-3066.

Lledé6 MD, Crespo MB, Fay ME, Chase MW. 2005. Molecular phylogenetics of Limonium and

related genera (Plumbaginaceae): biogeographical and systematic implications. Amer. J. Bot.

92: 1189-1198. http://dx.doi.org/ 10.3732/ajb.92.7.1189

Moharrek F, Kazempour Osaloo S, Assadi M. 2014. Molecular phylogeny of Plumbaginaceae

with emphasis on Acantholimon Boiss. based on nuclear and plastid DNA sequences in Iran.

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Mori Y, Sato Y, Takamatsu S. 2000. Evolutionary analysis of the powdery mildew fungi

using nucleotide sequences of the nuclear ribosomal DNA. Mycologia 92: 74-93.

http://dx.doi.org/10.2307/3761452

Song JG, Xu B, Zhang SH, Zhao ZY. 2015. First report of powdery mildew caused by

Erysiphe limonii on Acantholimon hedinii in China. Plant Dis. 99: 1038.

http://dx.doi.org/10.1094/PDIS-12-14-1339-PDN

Stamatakis A. 2006. RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with

thousands of taxa and mixed models. Bioinformatics 22: 2688-2690.

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Erysiphe acantholimonis sp. nov. (China) ... 629

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Microbiol. Sin. 20: 356-364.

MY COTAXON

July-September 2016— Volume 131, pp. 631-638

http://dx.doi.org/10.5248/131.631

Tubulicrinis martinicensis sp. nov.,

a corticioid species from Martinique (French West Indies)

GERALD GRUHN |, NILS HALLENBERG? & REGIS COURTECUISSE ”

' Office National des Foréts, Réseau mycologique, 5 avenue Mirandol, F-48000 Mende, France

’ Hessgade 35, DK-5500 Middelfart, Denmark

> Université de Lille (Laboratoire des sciences végétales et fongiques - LSVF),

B.P.83, F-59006 Lille Cedex, France

* CORRESPONDENCE TO: gerald.gruhn@onf.fr

ABSTRACT—The new species Tubulicrinis martinicensis, collected in Martinique Island

(French West Indies), is described and illustrated. The differences from other species with

cylindric cystidia with obtuse apex are discussed, and this new species is included in the key

to the species of Tubulicrinis known worldwide.

Key worps—Basidiomycota, Tubulicrinaceae, Hymenochaetales

Introduction

Mycological inventories initiated by R. Courtecuisse have been carried out

in French West Indies for several years on behalf of the French forestry office

(Office National des Foréts; Courtecuisse 2006, Voiry & Courtecuisse 2012,

Courtecuisse & Welti 2013). Several new species (primarily in Agaricomycetes

and Sordariomycetes) have been described in this project; here we describe the

first new corticioid species.

Tubulicrinis Donk is a morphologically well-delimited corticioid genus

belonging to the family Tubulicrinaceae Julich (Jiilich 1982) in the order

Hymenochaetales Oberw. (Frey et al. 1977). Its key character is the presence of

long cystidia with thick walls and multiple roots, dissolving in KOH (Hjortstam

et al. 1988, Bernicchia & Gorjon 2010) and often strongly amyloid (Larsson

2007). The type species, T’ glebulosus (Bres.) Donk, is non-typified (Hjortstam

2001) and closely related to T! gracillimus (Ellis & Everh. ex D.P. Rogers & HS.

Jacks.) G. Cunn.

632 ... Gruhn, Hallenberg & Courtecuisse

In this article, we describe as new and illustrate T. martinicensis, discuss its

similarities and differences to closely related species, and propose a modification

of the worldwide key to the genus by Hjortstam (2001).

Materials & methods

Macroscopic and microscopic studies were based on fresh and dried material.

Sections were prepared with a razor blade and observed in several solutions: ammoniacal

Congo Red, 3% KOH with additions of 1% phloxine B, Melzer’s, and Cotton Blue. The

dissolution time of cystidia was computed using 3% KOH. Measurements were done

on photos taken from the microscope under 1000x magnification, using Mycometre

software (Fannecheére 2011). In spore measurements the following format was used:

“N; (minimum-) d1-d9 (-maximum) um’, where “N” is the number of spores in the

sample and “d1l”and “d9” are the Ist and 9th decile measurements (Fannechére 2005).

Spore measurements were based on spores seen in profile and obtained from a spore

print (Duhem 2010), in Melzer's.

Specimens are conserved in the Herbarium, Département de Botanique, Université

de Lille, Lille, France (LIP); the Herbarium, Department of Biological and Environmental

Sciences, University of Gothenburg, Géteborg, Sweden (GB); and the private herbariums

of G. Gruhn (hb. Gruhn) and F Dammrich (hb. Dammrich). Additional material was

examined from the Herbarium, Université Claude Bernard, Lyon, France (LY) and from

the private herbariums of M. Gérard (hb. Gérard) and G. Trichies (hb. Trichies).

Tubulicrinis martinicensis G. Gruhn, Hallenb. & Courtec., sp. nov. Fies 1-3

MycoBAnk MB 815644

Differs from Tubulicrinis confusus by its narrower spores.

Type: French West Indies, Martinique, Les Anses d’Arlet, Grande Anse, Morne

Champagne, on deeply white-rotted wood of undetermined deciduous tree,

in a xerophilic coastal forest, 7 July 2012, leg. R. Courtecuisse (Holotype, LIP

GG-MARI12-206. Isotypes, hb. Gruhn MAR12-206; hb. Dammrich D11157; GB).

ETYMOLOGY: martinicensis epithet refers to the site of collection.

BASIDIOME annual, effused (30 x 10 cm), adnate, strongly attached to the wood,

cream-colored when fresh, irregularly cracked into polygons. Light ochraceous

and concolorous with the supporting wood when dry. HYMENIUM uneven when

fresh, smooth when dry. CONTEXT homogeneous, not stratified, concolorous

with the hymenium in the whole profile, 250-300 um thick, cracking into

pieces when dry. Margin thin and distinct, frequently floccose, concolorous

with the hymenium. SPORE PRINT white.

HYPHAL SYSTEM monomitic, context made up of a dense hyphal texture;

clamp connections present at all septa; hyphae thin-walled and smooth in

context but slightly thick-walled in subiculum, thin-walled in subhymenium,

2-3 um in diam., non amyloid. Lyocystip1a smooth, irregularly cylindrical,

distinctly sinuous in the upper part and rounded at apex, without crystals. Wall

Tubulicrinis martinicensis sp. nov. (Martinique) ... 633

Fic. 1. Tubulicrinis martinicensis (holotype, LIP GG-MAR12-206), fresh material.

Scale bar = 2 cm. Photo G. Gruhn.

thick (2-4 um) at the base and slightly amyloid. About two thirds of the outer

wall is irregularly inflated. Upwards, the wall becomes gradually thinner up

to the top and is quite thin along the apical third of the length (30-40 um).

Total dimensions: (60-)80-92(-112) x 7-7.5(-9) um, the wider part located

in the middle of the length. Near the apex sometimes slightly clavate with a

diameter of 4.5-5.5 um. Lumen usually symmetrical. CystTip1a originate deep

in the context, projecting up to 50-65 um above the level of the hymenium;

multiple roots have not been observed; external cystidial wall dissolving in 3%

KOH between 3 to 20 minutes. BAsip1A in a dense hymenium, short clavate,

9-12(-15) x 3.5-4 um, with one or several minor constrictions, thin-walled,

contents homogenous; sterigmata 4, thin, 3 um long, inamyloid. BasIDIOSPORES

subcylindrical to suballantoid, multiguttulate, N = 44, (5.1-)5.6-7.0(-7.1)

x (1.8-)1.9-2.6 um, Qm = 2.9; thin-walled and smooth, inamyloid, non-

dextrinoid, non-cyanophilous.

ADDITIONAL SPECIMENS EXAMINED:

Tubulicrinis gracillimus: FRANCE, VENDEg, [le d’Yeu, between St-Sauveur and

Port de la Meule,. on Quercus ilex, leg. M. Gérard, det. B. Duhem (hb.Gérard MG2440);

MOSELLE, commune de Moyeuvre, Forét Domaniale de Moyeuvre, on decorticated

Betula, leg. & det. G. Trichies (hb. Trichies GT00612). SWITZERLAND, BErw, Berner

Oberland, Grindelwald, on Juniperus sp., leg. & det. EF Dammrich (hb. Dammrich

D8312).

634 ... Gruhn, Hallenberg & Courtecuisse

Fic. 2. Tubulicrinis martinicensis (holotype, LIP GG-MAR12-206), hymenial details observed in

KOH-phloxine. A. mature and young basidia; B. spores C. cystidia.

Scale bars: A, C = 10 um; B = 5 um. Drawings G. Gruhn.

Tubulicrinis incrassatus: FRANCE, RHONE, St-Martin-en-Haut, lieu-dit le Bois de

Remayou, on Pinus sylvestris, leg. & det. G. Gruhn, conf. N. Hallenberg (hb. Gruhn GG-

151011-016). FRENCH WEST INDIES, GuapELOUPE, Basse-Terre, Carrére, on dead

stump, leg. & det. J. Boidin (LY JB-8106).

Discussion

Tubulicrinis martinicensis is well characterized by the shape of its long

tubular cystidia with obtuse apices and by its narrow slightly allantoid spores.

The new species is close to T. gracillimus, which shows long cylindrical

lyocystidia with a weak amyloid reaction. The shape of the lyocystidia is similar

Tubulicrinis martinicensis sp. nov. (Martinique) ... 635

Fic. 3. Tubulicrinis martinicensis (holotype, LIP GG-MARI12-206).

Cystidia observed in Melzer’s, with the horizontal line showing the emergence level.

Scale bar = 25 um. Drawings G. Gruhn.

636 ... Gruhn, Hallenberg & Courtecuisse

in both species, including a rounded apex without crystals, but in T. gracillimus

the thin-walled upper part is short, less than 20 um long (Hjortstam & Larsson

1988). Moreover, the spores of T: gracillimus are longer and narrower (7.5-8.5

x 1.75-2.0 um; Hjortstam & Larsson 1988).

Tubulicrinis confusus K.H. Larss. & Hjortstam is a rare species described

from Norway (Hjortstam & Larsson 1986). The cystidia are distinctly sinuous,

like those of T: martinicensis, but the diameters are smaller and become

narrower towards apex (Hjortstam et al. 1988), in contrast to the Martinique

species where the diameter widens towards the apex. Moreover, the spores of

T. confusus are ellipsoid and wider (2.8-3.5 um diam.; Hjortstam et al. 1988),

and its basidiome is odontioid in mature specimens.

Neither can T: martinicensis be confused with T: pseudoborealis Boidin &

Gilles, a tropical species known only from a single collection from Réunion

Island (Hjortstam & Ryvarden 2007). The spore dimensions are similar with

those of T: martinicensis, but the spore shape is ellipsoid to cylindrical. Also,

the thin-walled upper part of the lyocystidia is shorter and not sinuous, and the

basidiome is thin and porulose (Boidin & Gilles 1988).

Finally, the spores of T’ martinicensis are close to those of T’ incrassatus

Hallenb., which differs by its typically non-symmetrical cystidia (Hallenberg

1978, Hjortstam & Ryvarden 2007, Bernicchia & Gorjon 2010, Gruhn &

Rivoire 2016).

Key to Tubulicrinis incorporating T. martinicensis

Hjortstam (2001) published a key to known Tubulicrinis species, which we

propose to modify by inserting T. martinicensis [boldface] as follows:

244 (23b): Spores ellipsoid’or subcylindrigal 05.4 .see% tak oh ic® oe OE Ese 25

24b: Spores cyindricaltto-allantotd =. ..2.¢ 44, po gpegieed eo pew Saip edd tye bs Olde pean gee 0) 26

25a) (24a).:C ystidia distinotly-sintealis’ gg, seus ah pcg die picul gym bead dye DOE a pb sh ete a 34

D5b Gystidiaiotherwisen Reunions cfg yt ih ude ain nnce,eieee-ndeyal onder shea T. pseudoborealis

//BREAK//

30a (29b). Basidiomes thick, often cracking conspicuously ...................00. 31

S0bebasidiomies Chaninert cn kA Anak Anal Wyte Mita AR ete Lae, hanes Ba te ae 32

SLaABOaY SPOnes S57 x eB De tiny oe wis ae wo are ins Maree obec nase dee Tate tate 33

31b. Spores 6.0-9.0 x 1.5-2.25 um.

Common species, preferably on deciduous wood

(In its broad sense a cosmopolitan species. Note T. gracillimus,

which seems to be very close morphologically.) ................. T. glebulosus

Tubulicrinis martinicensis sp. nov. (Martinique) ... 637

32a (30b). Cystidia widened towards the apex, spores 6-8 x 1.5-1.8(-2) um.

Rather common in Northern Europe,

also fromi-North-Amertica: «24. 0s 4. end yng 24 dingee ¥en OP yng od tinge ee T. medius

32b. Cystidia with subcapitate apex, spores 5.5-9.5 x 1.5-1.8 um.

North America and Europe, rare species ............2..00 00 eee T. propinquus

33a (31a). Cystidia sinuous with symmetric lumen.

GAT DEAT Fy Sil oo he oe eh A TR 9 PU MRS oS eS T. martinicensis

33b. Cystidia otherwise, with typical asymmetric lumen.

Southern. Europe; Iran, Caribbean ok 008i. ite wo hae» sive aes T. incrassatus

34a (25a). Cystidia slightly acute, spores 2.8-3.5 um wide............... T. confusus

34b. Cystidia not acute, merely expanding at apex,

SPOLes <2; Se WISE ss auiita gs etoat gb sed Eset sa dnb et a mb eof aap pd T martinicensis

Acknowledgments

The authors thank Gérard Trichies, Maurice Gérard and Franck Dammrich for

loan of herbarium specimens. They also thank Dr Isabel Salcedo (UPV/EHU) and

Dr Genevieve Gates (UTAS) for peer reviewing. Collection of the type specimen has

been done during missions of the “Inventaire mycologique des Petites Antilles: diversité,

écologie et protection” headed by one of us since 2003 (RC). For these missions,

we thank the financial partners in 2012: Office National des Foréts (direction générale

et direction régionale Martinique) and DEAL-Martinique (Direction de lécologie, de

laménagement et du logement). The Société Mycologique de France is also thanked for

its help in the financial management of the field trips.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s. 1. Fungi Europaei 12. Ed. Candusso, Italia.

Boidin J, Gilles G. 1988. Basidiomycétes Aphyllophorales de I’ile de la Réunion X - Compléments

aux genres traités antérieurement (1" partie). Bulletin de la Société Mycologique de France

104(2): 59-72.

Courtecuisse R. 2006. Liste préliminaire des Fungi recensés dans les iles francaises des Petites

Antilles: Martinique, Guadeloupe et dépendances. I. Basidiomycétes lamellés et affines

(Agaricomycetideae s.l.). Documents Mycologiques 34(133-134): 81-140.

Courtecuisse R, Welti S. 2013. Liste préliminaire des Fungi recensés dans les iles francgaises des

Petites Antilles: Martinique, Guadeloupe et dépendances. II - Basidiomycétes non lamellés

(espéces gastéroides, rouilles et charbons exclus). Documents Mycologiques 35: 47-173.

Duhem B. 2010. Eléments pour une approche de létude des corticiés. I. La sporée. Bulletin de la

Société Mycologique de France 126(2): 143-160.

Fannechere G. 2005. Statistiques et notation des dimensions des spores. Bulletin de la Société

Mycologique de France 121(3-4): 255-292.

Fannechére G. 2011. Mycometre 2.05. http://mycolim.free.fr/sml/gener/logiciels1.html

Frey W, Hurka H, Oberwinkler F. 1977. Beitrage zur Biologie der niederen Pflanzen. Gustav Fischer

Verlag, Stuttgart. 233 p.

Gruhn G, Rivoire B. 2016. Tubulicrinis incrassatus (Basidiomycota), premiere récolte en France

métropolitaine. Bulletin de la Société Linnéenne de Lyon 85(7-8): 241-245.

638 ... Gruhn, Hallenberg & Courtecuisse

Hallenberg N. 1978. Wood fungi (Corticiaceae, Coniophoraceae, Lachnocladiaceae, Thelephoraceae)

in N. Iran. I. Iranian Journal of Plant Pathology 14: 38-87.

Hjortstam K. 2001. Tubulicrinaceae, a survey of the genera and species. Windahlia 24: 1-14.

Hjortstam K, Larsson KH. 1986. Notes on Corticiaceae (Basidiomycetes) XV. Some new species

from Northern Europe. Mycotaxon 26: 437-443.

Hjortstam K, Ryvarden L. 2007. Checklist of corticioid fungi (Basidiomycota) from the tropics,

subtropics, and the southern hemisphere. Synopsis Fungorum 22: 27-146.

Hjortstam K, Larsson KH, Ryvarden L. 1988. The Corticiaceae of North Europe. Phlebia,

Thanatephorus, Ypsilonidium. Fungiflora 8: 1450-1631.

Julich W. 1982 [“1981”]. Higher taxa of Basidiomycetes. Bibliotheca Mycologica. 85. 485 p.

Larsson KH. 2007. Re-thinking the classification of corticoid fungi. Mycological Research 111(9):

1040-1063. http://dx.doi.org/10.1016/j.mycres.2007.08.001

Voiry H, Courtecuisse R. 2012. Cing ans d’inventaire mycologique en forét de Martinique et de

Guadeloupe, premiers résultats. Rendez-vous Techniques ONF 80: 57-63.

MY COTAXON

July-September 2016— Volume 131, pp. 639-643

http://dx.doi.org/10.5248/131639

Roccella elisabethae with

lichenicolous Arthonia follmanniana in Turkey

MEHMET GOKHAN HALICI & ARIFE MERVE KAHRAMAN

University of Erciyes, Faculty of Science, Department of Biology, 38039 Kayseri, Turkey

CORRESPONDENCE TO: mghalici@gmail.com

AsBstTRACT—Roccella elisabethae and the lichenicolous fungus Arthonia follmanniana growing

on it are reported for the first time from Turkey. Comments on their habitats, substrata, and

key anatomical features are provided for each species along with habit photographs and

photographs of microscopic features of the Arthonia that have not previously been illustrated.

Key worps—Ascomycota, Besparmak Island, lichens, lichenized fungi, Mersin

Introduction

Studies on the lichenized and lichenicolous fungal biota of several regions in

Turkey have been recently increasing. Almost 1300 species of lichenized fungi

and 200 species of lichenicolous fungi are now known from the country (Halici

2008; Kocakaya et al. 2015, 2016). Still, there are many areas in Turkey that are

insufficiently studied or where no lichenological studies have been conducted.

There are few records of lichenized and lichenicolous fungi from the

province of Mersin (Halici et al. 2014, Halici: 2015). During a lichenological

exploration in the vicinity of the Akkuyu Nuclear Power Plant (Silifke, Mersin)

in the Eastern Mediterranean Region of Turkey, we made an excursion to

Besparmak Island located close to the power plant. On the vertical faces of

siliceous rocks in the highest points of the island, we found Roccella elisabethae

to be very common and frequently infected by Arthonia follmanniana; both

species are new records for Turkey.

Roccella elisabethae is known from the Atlantic coast of northwest Spain

(Galicia) and on islands in the warmer parts of the Atlantic Ocean (Azores,

640 ... Halici & Kahraman

Canary Islands, Cape Verde Islands, and Madeira; Aptroot & Schumm 2011,

Carballal 2013). Previously, Roccella phycopsis was the only Roccella species

known from Turkey. This subtropical genus of lichenized fungi is restricted

to coastal habitats and has 32 species worldwide (Tehler et al. 2010; Aptroot &

Schumm 2011).

The genus Arthonia includes both lichenized and lichenicolous fungi with

41 lichenicolous species previously reported from Turkey (e.g., Halici 2008).

The holotypes of five lichenicolous Arthonia species are from Turkey (Steiner

1899; Candan & Halici 2009; Halici & Candan 2009, 2011).

Material & methods

The specimens reported here are deposited in Erciyes University Herbarium,

Kayseri, Turkey (ERC). Examination was by standard microscopic techniques. Hand-

cut sections were studied in water, potassium hydroxide (KOH), and Lugol's solution

(I). Measurements were made in water. Ascospores were measured from five different

ascomata for each species. Measurements are given as minimum-maximum from

N measurements. The descriptions presented below are based on the Turkish specimens.

Taxonomy

Roccella elisabethae Tehler, Symb. Bot. Upsal. 34(1): 413. 2004 FIGS 1A4,B

THALLUS erect, sometimes pendent; branches terete, slightly flattened,

grey to slightly greenish, 1.5—-3 cm long, 0.5—0.8 mm wide. Cortex a palisade

plectenchyma, C+ red; medulla compact, whitish. SoRALIA present, laminal

sometimes terminal, C+ red. APOTHECIA and PYCNIDIA absent.

SPECIMEN EXAMINED: TURKEY, Ice1, Giilnar, Besparmak Island, 36°07’18”N

33°31'58’E, alt. 35 m, on vertical faces of siliceous rocks, 11 April 2012, M.G. Halica

(ERC AMEKA 0. 077).

ComMMENTS— The type specimen of R. elisabethae is from the Canary Islands

(Tehler et al. 2004), and the species is also known from other Atlantic Ocean

islands and the Atlantic coast of Spain (Aptroot & Schumm 2011, Carballal

2013). Our report from the eastern Mediterranean Sea therefore considerably

extends its range and R. elisabethae should be sought in other parts of the

Mediterranean.

Roccella phycopsis is morphologically similar to R. elisabethae but differs by

its C- soralia and its yellowish medulla (Tehler et al. 2004, Carballal 2013).

We note that the Turkish material of R. elisabethae has much shorter branches

compared with the 2-10 cm cited in the original description of the species,

which we attribute to the age of the specimen. Detailed descriptions and

illustrations of R. elisabethae are provided by Tehler et al. (2004), Aptroot &

Schumm (2011), and Carballal (2013).

Roccella elisabethae with Arthonia follmanniana (Turkey) ... 641

Fic. 1. Roccella elisabethae (ERC AMEKA 0.077): A, habitus; B, close view of soralia. Arthonia

follmanniana (ERC AMEKA 0.104): C, habitus; D, cross section of ascomata; E, ascospores

uniseriately arranged in asci; F, ascospores.

Arthonia follmanniana Diederich, Flechten Follmann: 180. 1995 FIGS 1C-F

ASCOMATA arthonioid, 0.1—0.3 mm diam, black, sometimes with a bluish

tinge due to superficial pruina, at first more or less immersed in the thallus

of the host lichen, becoming sessile, and mainly on the tips of the branches.

EPIHYMENIUM dark olivaceous to almost blackish; hypothecium pale to

dark brownish. HyMENIuM colourless, about 25 um tall; I+ red; K/I+ blue.

642 ... Halici & Kahraman

Ascli 8-spored; ascospores uniseriately arranged in the asci. ASCOSPORES

colourless, narrowly soleiform, mostly with a wide and shorter upper cell and

narrower and longer lower cell, 2—3-septate, 10—12(—13.5) x 3—4.5(—5) um

(N = 20).

SPECIMEN EXAMINED: TURKEY, Ice1, Giilnar, Besparmak Island, 36°07'18”N

33°31'58’E, alt. 35 m, on Roccella elisabethae on vertical faces of siliceous rocks, 11 April

2012, M.G. Halici (ERC AMEKA 0. 104).

CoMMENTS—The fungus occurred on the tips of the branches of Roccella

elisabethae. Arthonia follmanniana was described on R. galapagoensis in

Galapagos Islands (Diederich 1995) but is reported as common on R. canariensis

and R. fuciformis in the Canary Isles (Hafellner 1996). Roccella elisabethae is a

new host for this species, and the eastern Mediterranean is the third region

from which the species is known. A detailed anatomical description of Arthonia

follmanniana, illustrated with line drawings of the asci and ascospores, is

provided by Diederich (1995).

Acknowledgements

The manuscript was reviewed prior to submission by David L Hawksworth

(Universidad Complutense de Madrid, Spain) and Sergio Perez-Ortega (Real

Jardin Botanico (CSIC), Madrid, Spain). This study was financially supported by

FYL-2015-6298 coded project to Erciyes University.

Literature cited

Aptroot A, Schumm F. 2011. Fruticose Roccellaceae: an anatomical-microscopical atlas and

guide with a worldwide key and further notes on some crustose Roccellaceae or similar

lichens. 375 pp. Published by the authors and BoD [Books on Demand, available from

fschumm@online.de], The Netherlands.

Candan M, Halici MG. 2009. Two new lichenicolous Arthonia species from Turkey. Mycotaxon

107: 209-213. http://dx.doi.org/10.5248/107.209

Carballal R. 2013. El género “Roccella” en la Peninsula Ibérica y las Islas Baleares. Botanica

Complutensis 37: 13-20. http://dx.doi.org/10.5209/rev_bocm.2013.v37.42263

Diederich P. 1995. New or interesting lichenicolous fungi VI. Arthonia follmanniana, a new

species from the Galapagos Islands. 179-182, in: FJA Daniels et al. (eds). Flechten Follmann:

contributions to lichenology in honour of Gerhard Follmann. Botanical Institute, University

of Cologne.

Hafellner J. 1996. Bemerkenswerte Funde von Flechten und lichenicolen Pilzen auf makaronesischen

Inseln V. Uber einige Neufunde und zwei neue Arten. Herzogia 12: 133-145.

Halici MG. 2008. A key to the lichenicolous Ascomycota (including mitosporic fungi) of Turkey.

Mycotaxon 104: 253-286.

Halici1 MG. 2015. New records of crustose Teloschistaceae and lichenicolous fungi from Turkey.

Mycotaxon 130: 769-773. http://dx.doi.org/10.5248/130.769

Halici1 MG, Candan M. 2009. New lichenicolous fungi from Turkey. Nova Hedwigia 88: 483-490.

http://dx.doi.org/10.1127/0029-5035/2009/0088-0483

Roccella elisabethae with Arthonia follmanniana (Turkey) ... 643

Halici MG, Candan M. 2011. Arthonia anatolica sp. nov. (Arthoniaceae) on Aspicilia contorta

subsp. hoffmanniana, a new lichenicolous species from Turkey. Mycotaxon 116: 335-339.

http://dx.doi.org/10.5248/116.335

Halici MG, Vondrak J, Demirel R, Ceylan A, Candan M. 2014. Teloschistaceae (lichenized

ascomycetes) on Turkey II. - Some poorly known taxa, supported by molecular data. Nova

Hedwigia 98: 449-458. http://dx.doi.org/10.1127/0029-5035/2013/0163

Kocakaya Z, Halic1 MG, Kocakaya M. 2015. Phoma candelariellae sp. nov., a lichenicolous fungus

from Turkey. Mycotaxon 130: 1185-1189. http://dx.doi.org/10.5248/130.1185

Kocakaya M, Halici MG, Pino-Bodas R. 2016. New or additional cladoniicolous fungi for Turkey.

Turkish Journal of Botany 40: 308-311. http://dx.doi.org/10.3906/bot-1502-8

Steiner J. 1899. Lichenes. Denkschriften der Kaiserlichen Akademie der Wissenschaften, Wien.

Mathematisch-Naturwissenschaftliche Classe 68: 222-238.

Tehler A, Dahlkid U, Eldenas P, Feige GB. 2004. The phylogeny and taxonomy of Macaronesian,

European and Mediterranean Roccella (Roccellaceae, Arthoniales). Symbolae Botanicae

Upsalienses 34(1): 405-428.

Tehler A, Irestedt M, Wedin M, Ertz D. 2010. The Old World Roccella species outside Europe and

Macaronesia: taxonomy, evolution and phylogeny. Systematics and Biodiversity 8: 223-246.

http://dx.doi.org/10.1080/14772001003789554

MY COTAXON

July-September 2016— Volume 131, pp. 645-652

http://dx.doi.org/10.5248/131.645

Diaporthe henanensis sp. nov.,

an endophytic fungus in Ziziphus jujuba from China

YI YANG *, YU-XIA Guo *, YA-KUN ZHANG,

HAI-YAN Wu & MENG ZHANG *

Henan Agricultural University, 95 Wenhua Road, Zhengzhou, Henan, 450002, China

* CORRESPONDENCE TO: Z2006@126.com

ABSTRACT—A species of Diaporthe isolated from fruit of Ziziphus jujuba during a study of the

diversity of endophytic fungi in Zhengzhou, Henan, China, was found to be morphologically

and molecularly distinct from known species. This new species, proposed as Diaporthe

henanensis, differed from other Diaporthe species by its pyriform, relatively wide y-conidia.

Analysis of multi-locus DNA sequences (ITS, TUB, HIS) showed that D. henanensis was

closely related to D. nobilis, D. neilliae, and D. alnea.

Key worps—endophytes, Phomopsis, Ascomycota, systematics

Introduction

Diaporthe (= Phomopsis) species are common plant pathogens, endophytes

or saprobes (Webber & Gibbs 1984, Carroll 1986, Boddy & Griffith 1989,

Rehner & Uecker 1994, Udayanga et al. 2011). In recent years there has been an

acceleration in the discovery and naming of new Diaporthe species recognized

primarily through molecular analyses (Santos & Phillips 2009; Santos et al.

2011; Crous et al. 2011; Udayanga et al. 2012, 2014).

Udayanga et al. (2011) suggested Diaporthe species should be identified

by a combination of molecular, morphological, cultural, pathological and

mating type data. Tan et al. (2013) reported six new Diaporthe species from

Australia through ITS, TUB, and TEF gene sequence analyses. Huang et al.

(2013) reported two Diaporthe species based on ITS, EF, TUB, and CAL gene

* These authors contributed equally to this study.

646 ... Yang, Guo & al.

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Diaporthe henanensis sp. nov. (China) ... 647

sequence analyses. Gomes et al. (2013) investigated 243 isolates of Diaporthe,

divided them into 95 species using multi-locus DNA sequence data (incl. ITS,

HIS, TUB, CAL, TEF1), and concluded that future species descriptions should

include at least ITS as well as HIS or TUB sequence data.

Ziziphus jujuba Mill. (Rhamnaceae) is a small deciduous tree grown in

China for its edible fruit. During a study of the diversity of endophytic fungi in

Zhengzhou, Henan, China, a fungus regularly isolated from fruit of Z. jujuba

was shown to represent a novel species of Diaporthe, which is described here as

Diaporthe henanensis.

Materials & methods

Isolates, media, and culturing

Healthy fruit of Ziziphus jujuba was collected near Zhengzhou, Henan Province,

China. Tissue pieces of about 3 mm? were treated with 75% ethanol for 30 s, surface

disinfected in 3% sodium hypochlorite for 3 min, rinsed with sterile distilled water,

then placed on potato carrot agar (PCA) plates at 25 °C for one week. Young colonies

were transferred to potato dextrose agar (PDA) plates and kept at room temperature

under alternating light and dark periods (Xie et al. 2010) for conidium production.

Morphological characteristics of conidia and conidiophores were microscopically

observed after 10 d.

The type specimen was deposited in the Herbarium Mycologicum Academae Sinicae,

Beijing, China (HMAS), and cultures were deposited in China General Microbiological

Culture Collection Centre, Beijing, China (CGMCC) and in the Herbarium Henan

Agricultural University: Fungi, Zhengzhou, China (HHAUF).

DNA extraction, polymerase chain reaction, and sequencing

Total DNA was extracted from mycelium scraped from the surface of a PDA plate

with the Cell Genomic DNA Isolation Kit (LifeFeng, Shanghai, China) following the

manufacturer's instructions. The primers ITS1 and ITS4 (White et al. 1990) were used

to amplify the ITS1-5.8S-ITS2 rDNA region (ITS) (Luo & Mitchell 2002). The primers

T1 (O'Donnell & Cigelnik

1997) and bt2b (Glass & Donaldson 1995) were used to amplify part of the tubulin gene

(TUB). The primers CYLH3F (Crous et al. 2004) and H3-1b (Glass & Donaldson 1995)

were used to amplify part of the histone H3 gene (HIS). PCR was carried out using

an Eppendorf PCR thermal cycler (Germany). The reaction products were analyzed by

electrophoresis on a 1.0% agarose gel in tris-acetate-EDTA (TAE) buffer with ethidium

bromide (0.1 mg/mL) and visualized under UV light (Xie et al. 2010). The PCR products

were sequenced by Shanghai Sangon Biological Engineering Technology & Service Co.,

Ltd., China.

The ITS sequences were initially aligned with published ITS sequence data of

Diaporthe spp. obtained from the NCBI using ClustalW in MEGA ver. 5.2 (Tamura et al.

2011). Diaporthella corylina was selected as the outgroup taxon. A neighbor-joining (NJ)

648 ... Yang, Guo & al.

analysis using the Kimura-2 parameter with Gamma distribution was applied to nine

similar species and the closest phylogenetic neighbours were selected for a combined

analysis using ITS, TUB and HIS genes (TABLE 1). A Maximum likelihood tree using

the combined dataset was generated in MEGA 5.2 using the Tamura-Nei substitution

model with Gamma distribution. Bootstrap support values with 1000 replications were

calculated for tree branches (Tan et al. 2013).

090 OOP

2 00

999 9 9

Fic. 1 Diaporthe henanensis on PDA.

A. a-conidia; B. y-conidia; C. Conidiophores; D. B-conidia.

Scale bar = 10 um.

Diaporthe henanensis sp. nov. (China) ... 649

Taxonomy

Diaporthe henanensis Yi Yang, H.Y. Wu & Meng Zhang, sp. nov. FIG.1

MycoBank MB 804356

Differs from all other Diaporthe spp. by its y-conidia that are broadly pyriform and by

its multi-locus gene sequence data.

Type: China, Henan Province, Zhengzhou, from healthy fruit of Ziziphus jujuba,

26 Sep. 2012, coll. Y. Yang (Holotype, HMAS246233 [dried culture]; ex-type strain,

CGMCC3.17639 = HHAUF12291; GenBank KC898258, KF600608, KF600609).

EtymMoLoecy: Named for the collection locality.

Pycnidia scattered in PDA, dark brown to black, globose to subglobose,

800-1700 um diam. Conidiophores subcylindrical, hyaline, smooth, simple

or branched, 14-18 x 1.5-2.0 um. Conidiogenous cells phialidic, cylindrical,

tapering towards the apex, with periclinal thickening. Collarettes inconspicuous.

Three kinds of conidia were produced: a-conidia, unicellular, hyaline elliptical

or clavate, one end obtuse, the other acute, 2-guttulate, 5-8 x 2-3 um; B-conidia

hyaline, aseptate, without guttules, filiform, curved or hamate, with obtuse

ends, 15.5-31 x 0.8-1.5 um; y-conidia, pyriform, 5-7 x 3.5-5.0 um.

90 Diaporthe alleghaniensis CBS 495.72

99 Diaporthe juglandina CBS 121004

Diaporthe vaccini’ GBS 122114

59 100! Diaporthe vaccinii CBS 122115

Diaporthe celastrina CBS 139.27

_ Diaporthe eres CBS 375.61

99! Diaporihe eres CBS 445.62

s9 7 Diaporthe cf. nobilis RG-2013 CBS 116953

ont Diaporthe cf nobilis RG-2013 CBS 338.89

si Diaporthe henanensis HHAUF12291

100' Diaporthe henanensis HHAUF12292

= Diaporthe neiliae CBS 144.27

~499| | Draporthe ainea CBS 146.46

400 Diaporthe alnea CBS 159.47

Diaporthe gardeniae CBS 288.56

Diaporthella corylina CBS 121124

0.02

Fic. 2 Maximum likelihood tree inferred from combined analysis of three genes (ITS, TUB, HIS).

The percentage of replicate trees in which the associated taxa clustered together in the bootstrap

test (1000 replicates) is shown next to the branches. The tree was rooted to Diaporthella corylina.

The newly sequenced strains of our new species are in bold..

650 ... Yang, Guo & al.

ADDITIONAL MATERIAL EXAMINED: CHINA, HENAN PROVINCE, Zhengzhou, from

healthy fruit of Ziziphus jujuba, 26 Sep. 2012, coll. Y. Yang (HHAUF12292; GenBank

KC898259, KF600610, KF600611).

Discussion

All other Diaporthe species differ from D. henanensis either by lacking

y-conidia or by producing y-conidia that are not broadly pyriform (Crous et al.

2012, Gomes 2013, Uecker 1988, Qi et al. 2007). The initial ITS sequence data

support D. nobilis, D. neilliae, D. alnea, D. juglandina, D. eres, D. gardeniae,

D. alleghaniensis, and D. vaccinii as the most closely related phylogenetic

neighbours. The two isolates clustered with each other in a single clade with

100% bootstrap support in the ML tree inferred from analysis of three genes

(ITS, TUB, HIS; Fia. 2).

Many species of Diaporthe are known to be endophytic on a range of host

plants (Murali et al. 2006, Chaeprasert et al. 2010, Rocha et al. 2011, Sun et al.

2011). In this study, D. henanensis was isolated and described as new based on

its distinctive morphology (broadly pyriform y-conidia) and multi-locus gene

sequence data (ITS, TUB, HIS).

Acknowledgments

We thank Dr R. E. Castafieda Ruiz, Instituto de Investigaciones Fundamentales en

Agricultura Tropical (INIFAT), Cuba and Dr R. G. Shivas (Queensland Plant Pathology

Herbarium, Australia) for pre-submission review. This project was supported by the

National Natural Science Foundation of China (31171804 and 30970016) and Program

for Science & Technology Innovation Talents in Universities of Henan Province

(13HASTIT007).

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Crous PW, Groenewald JZ, Shivas RG, Edwards J, Seifert KA, et al. 2011. Fungal Planet description

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Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR

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Gomes RR, Glienke C, Videira SIR, Lombard L, Groenewald JZ, Crous PW. 2013. Diaporthe:

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Luo GZ, Mitchell TG. 2002. Rapid identification of pathogenic fungi directly from

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Murali TS, Suryanarayanan TS, Geeta R. 2006. Endophytic Phomopsis species: host range

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Qi PK, Jiang ZD, Xiang MM. 2007. Flora Fungorum Sinicorum Vol. 34 Phomopsis. Science

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Rehner SA, Uecker FA. 1994. Nuclear ribosomal internal transcribed spacer phylogeny and

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MY COTAXON

July-September 2016— Volume 131, pp. 653-661

http://dx.doi.org/10.5248/131.653

Notes on rust fungi in China 1.

Autoecious life cycle of Puccinia tatarinovii on Prenanthes

JING-XIN Jr!, QI WANG 1, ZHUANG L1?, Yu Lr & MAKOTO KAKISHIMA !*"

' Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun, Jilin 130118 China

College of Plant Protection, Shandong Agricultural University, Taian 271000 China

> University of Tsukuba, Tsukuba, Ibaraki 305-8572 Japan

* CORRESPONDENCE TO: kakishima.makoto.ga@u.tsukuba.ac.jp

ABsTRACT—During investigations of rust fungi in Jilin Province, northeast China, aecial

and telial stages were observed on Prenanthes tatarinowii in different seasons. The telial

stage was identified as Puccinia tatarinovii by morphological observations. Life cycle

connection between aecial and telial stages was clarified by field observations and molecular

data. Morphology of the spermogonial and aecial stages is described based on specimens

connected to telial stage. Lectotype and epitype specimens are designated.

KEY worps—aecial stage, ecology, morphology, spermogonium, taxonomy

Introduction

Puccinia tatarinovii was described by Tranzschel (1939) based on four

specimens on Prenanthes tatarinowii Maxim. (Compositae) collected in the Far

East of Russia and northeast of China (Manchuria). Although aecial, uredinial

and telial stages were reported in this description no morphological characters

of the aecial stage were described, and no type specimen was designated.

Puccinia tatarinovii is morphologically similar to Pu. orbicula Peck & Clinton

recorded on Prenanthes spp., although verrucae on the surface of its teliospores

are larger than those of Pu. orbicula (Tranzschel 1939, Ito 1950, Hiratsuka

et al. 1992, Zhuang 2003). In China, telia of a rust fungus on Pr. tatarinowii

were collected at Longtan mountain, Jilin, Jilin Province, northeast China in

September 1923 and were reported as Puccinia prenanthis-racemosae P. Syd. &

Syd. [= Pu. orbicula] (Miura 1928). Later, the rust specimens on Pr. tatarinowii

were mostly treated as Pu. tatarinovii (Ito 1950, Tai 1979, Wei & Wang 1986,

654 ... Ji&al.

Zhuang 2003). However, they were sometimes reported as Pu. orbicula because

they are morphologically similar and their host plants are in the same genus,

Prenanthes (Tai 1979). Both rust species have also been reported to have an

autoecious life cycle (Tranzschel 1939, Ito 1950, Cummins 1978, Zhuang

2003), but spermogonial and aecial stages of Pu. tatarinovii have not been

clearly distinguished, and they have been confused with those of Pu. orbicula

as these stages have not been found on the same plants at the same time. ‘This

is also largely because no morphological information has been recorded for the

aecia of Pu. tatarinovii, and there is no record of aecial specimens in China,

though many specimens of this species have been collected on Pr. tatarinowii

and Pr. macrophylla Franch. from various areas of China (Wei & Wang 1986,

Zhuang 2003). Azbukina (2005) described spermogonial and aecial stages on

Pr. tatarinowii, but connection between aecial and telial stages of this species

was not demonstrated.

During our investigation of rust fungi in Jilin Province, China, we found

aecial (Fic. 1A) and telial stages (Fic. 2A) on Pr. tatarinowii at the same place in

Jilin city, although they were observed at different times of the year. Therefore,

we suspected that these stages were produced by the same species. We carried

out morphological observations of these stages and also molecular analyses to

confirm the connection between the aecial and telial stages. We report here the

autoecious life cycle of Pu. tatarinovii and describe its morphology based on

these specimens. We also designate an epitype specimen for this species.

Materials & methods

Specimens

Aecial and telial specimens on Pr. tatarinowii were collected in Hongyegu, Jiaohe,

Jilin city, Jilin Province, China (43°42’13”N 127°04’18”E, alt. 537 m) in June and

September, 2015, and used for morphological observations and molecular analysis.

Specimens identified as Pu. tatarinovii and deposited in the Fungal Herbarium,

Institute of Microbiology, Academia Sinica (HMAS), were borrowed for comparative

morphology. Dry specimens used in the experiments were deposited in the Herbarium

of Mycology, Engineering Research Center of Chinese Ministry of Education for Edible

and Medicinal Fungi, Jilin Agricultural University, China (HMJAU).

Morphological observations

Light microscopy (LM) was used to examine morphological characters including

the size and shape of sori and spores. Spores or thin-sections of sori from specimens

were mounted in a drop of lactophenol solution on glass slides for LM. Approximately

50 spores from each specimen were randomly chosen and the length, width, and wall

thickness of spores were measured using Leica LAS X software attached to a Leica

DM2000 microscope (Leica, Germany).

The surface features of spores were examined by scanning electron microscopy

(SEM). For SEM, samples obtained from dry specimens were attached to specimen

Puccinia tatarinovii on Prenanthes (China) ... 655

holders by double-sided adhesive tape and coated with platinum-palladium using a

Hitachi MC1000 Ion Sputter Coater and examined with a Hitachi SU8010 SEM operated

at 5 kV.

DNA extraction and sequencing of rDNA-ITS regions

The total genomic DNA was extracted from about 200 spores obtained from a single

sorus (aecidium or telium) on specimens. Spores were crushed between two sterilized

glass slides and suspended in 30 ul extraction buffer [10mM Tris-HCl pH 8.3, 1.5 mM

MgCl, 50 mM KCL, 0.01% sodium dodecyl sulfate (SDS), 0.01% Proteinase K], and the

suspensions were incubated at 37°C for 1 hour and 95°C for 10 min, followed by a 4°C

soak (Suyama et al. 1996, Virtudazo et al. 2001). From the crude extract, 5-7 ul samples

were used directly for each polymerase chain reaction (PCR). The rDNA-ITS region

was amplified using primers ITSIF (5’-cTTGGTCATTTAGAGGAAGTAA-3’) (Gardes &

Bruns 1993) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’) (White et al. 1990). The PCR

amplifications were performed in 50 ul of mixture containing 5 ul of template DNA, 200

ul of each primer, 25 ul of Premix Taq™ (TaKaRa Taq™ Version 2.0 plus dye) (TaKaRa,

Tokyo, Japan), and 18 pl of ddH,O. Cycling conditions for amplification consisted of

94°C for 5 min, followed by 35 cycles of denaturation at 94°C for 30 s, annealing at

45°C for 30 s and extension at 72°C for 1 min, and a final extension at 72°C for 10

min. PCR products were separated on 1% agarose gels that pulsed Nucleic Acid Stain

(Beijing Dinggou Changsheng Biotechnology Co. Ltd.) and purified using the TaKaRa

MiniBEST Agarose Gel DNA Exaction Kit Ver.4.0 (TaKaRa, Tokyo, Japan). Purified

PCR products were cloned in pEASY -T1 Cloning Vector (Transgene Biotech, Beijing,

China) and then transferred into Trans1-T1 phage, resistant chemically competent cell

according to the manufacturer's instructions. The positive clones were sequenced by

Sangon Biotech Co., Shanghai, China.

Sequence similarity (%) of rDNA-ITS regions among specimens was calculated by

the NCBI program (NCBI, USA).

Results & discussion

Life cycle

Sequences obtained from specimens of aecial stage with spermogonial

(HMJAU8206) and telial stages (HMJAU8204) were used for analysis and these

data were deposited at GenBank. However, DNA could not be extracted from

spores obtained from the HMAS specimens. Chemical treatment of specimens

in the herbarium may affect their DNA. Sequence similarity of rDNA-ITS

regions (689 bp in length) between aecial and telial specimens was 100%.

Therefore, both specimens are genetically identical and life cycle connection

between aecial and telial stages on Pr. tatarinowii was confirmed. In the Jilin

Province field observation, the uredinial stage was not found, although a few

urediniospores were recognized in most specimens deposited in HMAS. We

suspect that production of the uredinial stage is affected by climatic conditions.

From the results of DNA analysis and specimen observation, the

autoecious macrocyclic life cycle of Pr. tatarinowii was clarified. Inoculations

656 ... Ji &al.

of plants with spores are best for clarifying life cycles, but collections of the

plant are restricted on the mountain because of nature conservation and its

transplantation to distant places is difficult. Therefore, we used DNA analysis

instead of inoculations and confirmed that sequence analysis is a useful tool to

clarifying a life cycle. Tranzschel (1939) and Azbukina (2005) reported aecia,

but provided no information that these aecia were observed on the same host

plants that produced telia. Therefore, our results provide the first evidence of

the complete life cycle in this species.

ee he

+ Jaze Th t “5

FiGurE 2. Uredinial and telial stages of Puccinia tatarinovii on Prenanthes tatarinowii:

A, B. Telia on the upper (A) and under (B) leaf surface of Pr. tatarinowii; D. Vertical section

of a telium; D. Teliospores; E. Urediniospores; F. Vertical section of an uredinium. Scale

bars: C = 40 um; D, F = 30 um; E = 20 um.

Puccinia tatarinovii on Prenanthes (China) ... 657

Morphology

Based on observations with specimens collected in Jilin Province (HMJAU)

and specimens borrowed from HMAS, an updated description of Pu. tatarinovii

is provided.

FiGure 1. Spermogonial and aecial stages of Puccinia tatarinovii on Prenanthes tatarinowii:

A, B. Pr. tatarinowii producing aecia in June 2015; C. Aecia produced on the stem; D. Vertical

section of a spermogonium (Type 4); D. Aeciospores; E. Vertical section of an aecium.

Scale bars: C, D = 20 um; E = 30 um.

Puccinia tatarinovii Kom. & Tranzschel, Consp. Ured. URSS: 393, 1939. Figs 1-3

TYPE: Russia, Russian Far East, Primorsky Territory, near Vladivostok, II on Prenanthes

tatarinowii, 10 Sep. 1913, leg. N. Djukina (Lectotype designated here, LE44326). China,

Jilin, 0, I on Pr. tatarinowii, 24 June 2015, leg. M. Kakishima & J.X. Ji (Epitype designated

here, HMJAU8206; GenBank KU955596).

Spermogonia subepidermal, Group V (type 4). Aecia mainly caulicolous,

sometime hypophyllous, in small roundish groups or along veins, short,

cupulate; peridia yellowish white, the margin revolute, lacerate, peridial cells

658 ... Ji&al.

polygonal or rhomboid, the outer walls striate, the inner walls verrucose, thin;

aeciospores angular-globose or ellipsoid, yellowish, 13.9-22.4 x 11.4-18.0 um

(av. 17.1 x 15.2 um), walls hyaline and verrucose, 0.3-1.6 um (av. 0.9 um) thick.

Uredinia amphigenous, scattered, orange-yellow, globoid; urediniospores

globoid or ellipsoid, 19.0-28.6 x 17.6-25.3 um (av. 24.6 x 22.5 um), walls

uniformly thick 1.0-3.0 um (av. 1.8 um), echinulate. Telia mainly hypophyllous

or rarely epiphyllous, especially in heavy infections, subepidermal, erumpent

when mature, brown to dark brown; teliospores 2-celled, borne singly on

pedicels, fusiform or oblong, sub-rotund or narrowed above, more or less

constricted at the septum 25.8-44.3 x 22-32.2 um (av. 34.1 x 23.8 um), walls

pigmented, yellowish-brown, 0.5-3.3 um (av. 1.9 um) thick, largely verrucose;

pedicels up to 5.2 um long.

Hosts & DISTRIBUTION — On Prenanthes tatarinowii: Russia: Primorsky

Territory; China: Jilin, Hubei, Beijing, Hebei, Shaanxi, Sichuan (Tranzschel

1939, Zhuang 2003, Azbukina 2005). On Pr. macrophylla: China: Sichuan

(Zhuang 2003).

ADDITIONAL SPECIMENS EXAMINED —

0, I on Pr. tatarinowii: CHINA, SICHUAN, 17 Sep. 1989 (HMAS63916); HUBEI, 19

Aug. 1984 (HMAS50409); 27 Aug. 1984 (HMAS50414).

II on Pr. tatarinowii: CHINA, BEIJING, 8 June 1957 (HMAS55746); HuBEI, 4 July

1984 (HMAS57292); SHAANXI, 9 July 1988 (HMAS64179, HMAS64181).

II, III on Pr. tatarinowii: CHINA, Huet, 18 Aug. 1984 (HMAS50410); 19 Aug.

1984 (HMAS50411); 27 Aug. 1984 (HMAS50413); 29 Aug. 1984 (HMAS57289).

III on Pr. tatarinowii: CHINA, JILIN, 2 Oct. 2015 (HMJAU8201); 23 Sep. 2015

(HMJAU8202, HMJAU8203); 21 Sep. 2015 (HMJAU8204; GenBank KU950447);

15 Sep. 2013 (HMJAU8207); Huse, 27 Aug. 1984 (HMAS50412); HEBEI, 1936

(HMAS06987); Aug. 1936 (HMAS12027); BEIJING, 3 Sep. 1957 (HMAS65837); 1 Aug.

1957 (HMAS65838); 5 Oct. 1957 (HMAS25656); SICHUAN, 23 Sep. 1989 (HMAS63915,

HMAS63914, HMAS63931, HMAS63932, HMAS63933); 17 Sep. 1989 (HMAS63934,

HMAS63935, HMAS63936).

III on Pr. macrophylla: CHINA, S1cHuAN, 13 Oct. 1989 (HMAS63917); 14 Oct.

1989 (HMAS63918, HMAS63919); 17 Sep. 1989 (HMAS63920, HMAS63921).

The morphological characters of uredinial and telial stages of Pu. tatarinovii

on Pr. tatarinowii were similar to the descriptions of Pu. tatarinovii reported

by Tranzschel (1939), Ito (1950), Wei & Wang (1986), Zhuang (2003), and

Azbukina (2005), although there is some variation in teliospore size among

these descriptions. Uredinial and telial morphologies of Pu. tatarinovii are

quite similar to Pu. orbicula except for the surface structure of teliospores

(Tranzschel 1939, Ito 1950, Hiratsuka et al. 1992, Zhuang 2003). Verrucae on

teliospores of Pu. tatarinovii are larger than those of Pu. orbicula as reported by

Tranzschel (1939) and Zhuang (2003). Therefore, verrucal size is an important

character for distinguishing these species. Our observations of the aecial stage of

Puccinia tatarinovii on Prenanthes (China) ... 659

FiGure 3. Aecial, uredinial, and telial stages of Puccinia tatarinovii on Prenanthes tatarinowii

observed with SEM: A. Aecidium; B. Aeciospore with verrucose surface; C. Telium;

D. Teliospores with large verrucae; E. Uredinium; F. Urediniospore with echinulate surface.

Scale bars: A = 200 um; B = 0.5 um; C = 100 um; D = 20 um; E = 50 um; F = 8 um.

Pu. tatarinovii showed that its aeciospores are smaller than those of Pu. orbicula

(23-30 x 18-22 um) as also reported by Cummins (1978). Our observations

are also similar to the description of Azbukina (2005). Therefore, size of

aeciospores is an important character to recognize species. Puccinia tatarinovii

is recorded on only two host plants, Pr. tatarinowii and Pr. macrophylla, and

660 ... Ji &al.

distributed only in China and the Russian Far East (Tranzschel 1939, Zhuang

2003, Azbukina 2005), whereas Pu. orbicula is widely distributed on species of

Prenanthes (Cummins 1978, Azbukina 1984, 2005, Hiratsuka et al. 1992).

Tranzschel (1939) did not designate a type specimen of Pu. tatarinovii

although he listed four specimens in his description. Petrak (1950: 92) also did

not indicate the type for this species. Later, Azbukina (2005) selected a lectotype

from these specimens but did not designate it according to the requirements of

ICN Art. 7.10 (McNeill et al. 2012). Therefore, we checked specimens deposited

in LE and selected as lectotype the specimen which Tranzschel separated from

others as a type (personal communication from LE). Additionally, we designate

a spermogonial and aecial specimen as an epitype, because aecial morphology

is important to recognize the species.

Acknowledgments

This work was financed by the Science and Technique Foundation of Education

Department of Jilin Province (20130206073NY) and Recruitment Program of Foreign

Experts (WQ20122200064). We wish to thank Dr. E.H.C. McKenzie (Landcare

Research, Auckland, New Zealand) and Dr. C.M. Denchev (Bulgarian Academy of

Sciences, Sofia, Bulgaria) for critical reading of the manuscript and suggestions. We

express our thanks to Dr. O. Morozova (Komarov Botanical Institute, St. Petersburg,

Russia (LE)) for the examination of specimens. We also thank Dr. Y.-J. Yao, curator of

the Fungal Herbarium (Institute of Microbiology, Academia Sinica, Beijing, HAMS)

for loan of specimens. We also thank Dr. B. Qu (Shenyang Agricultural University) for

identification of plant specimens.

Literature cited

Azbukina ZM. 1984. The manual of rust fungi in Soviet Far East. Nauka, Moscow.

Azbukina ZM. 2005. Rust fungi. Cryptogamic plants, fungi and mosses of the Russian Far East, vol.

5. Dalnauka, Vladivostok. 616 p. (In Russian)

Cummins GB. 1978. Rust fungi on legumes and composites in North America. University of

Arizona Press, Tucson.

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes:

application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

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.

McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp

S, Marhold K, Prado J, Prud’homme van Reine WF, Smith GF, Wiersema JH, Turland NJ. 2012.

International Code of Nomenclature for algae, fungi, and plants (Melbourne Code). Regnum

Vegetabile 154. http://www.iapt-taxon.org/nomen/main.php

Miura M. 1928. Flora of Manchuria and east Mongolia II. Cryptogams, Fungi.

Minamimanshutetsudo, Dalian.

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Petrak FE 1950. List of new species and varieties of fungi, new combinations and new names

published 1936-1939. Petrak’s Lists 8. 117 p.

Suyama Y, Kawamuro K, Kinoshita I, Yoshimura K, Tsumura Y, Takahara H. 1996. DNA sequence

from a fossil pollen of Abies spp. from Pleistocene peat. Genes & Genetic Systems 71: 145-149.

http://dx.doi.org/10.1266/ggs.71.145

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Peking.

Tranzschel W. 1939. Conspectus uredinalium URSS. Academy of Sciences USSR, Moscow. 426 p.

Virtudazo EV, Nakamura H, Kakishima M. 2001. Phylogenetic analysis of sugarcane rusts based

on sequences of ITS, 5.8 S rDNA and D1/D2 regions of LSU rDNA. Journal of General Plant

Pathology 67: 28-36. http://dx.doi.org/10.1007/PL00012983

Wei SX, Wang YC. 1986.Taxonomic studies of Puccinia on Compositae in China. Acta Mycologica

Sinica Suppl. 1: 185-226.

White TJ, Bruns TD, Lee SB, Taylor JW. 1990. Amplification and direct sequencing of

fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds.).

PCR protocols: a guide to methods and applications. San Diego, CA. Academic Press.

http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

Zhuang JY. 2003. Flora Fungorum Sinicorum vol. 19, Uredinales (II). Science Press, Beijing.

MYCOTAXON

July-September 2016— Volume 131, pp. 663-670

http://dx.doi.org/10.5248/131.663

Gymnopus ramulicola sp. nov.,

a pinkish species from southern China

SHU-FANG DENG !”, Ta1-Hut Lt !?", Z1-DE JIANG ! & BIN SONG ?

' Department of Plant Pathology/Guangdong Province Key Laboratory of Microbial Signals &

Disease Control, South China Agricultural University, Guangzhou 510642, China

*State Key Laboratory of Applied Microbiology Southern China,

Guangdong Provincial Key Laboratory of Microbial Culture Collection & Application,

Guangdong Institute of Microbiology, Guangzhou 510070, China

“ CORRESPONDENCE TO: mycolab@263.net

ABSTRACT—Gymnopus ramulicola, a new species of G. subsect. Vestipedes, is described from

China. It is characterized by a pale red to pinkish white pileus, adnate to subfree lamellae,

a (sub)insititious stipe with white mycelium at an enlarged base, a simple pileipellis cutis

with occasional weakly diverticulate elements, abundant clamp connections, and habitat on

twigs or sticks of broadleaved trees. The new species is compared with similar species in its

subsection, and its phylogenetic position is inferred by a maximum likelihood tree based on

ITS sequences.

Key worps—Omphalotaceae, phylogeny, taxonomy

Introduction

During several macrofungal field surveys in southern China, three

interesting gymnopoid collections were obtained from Hainan and Jiangxi

provinces. Morphological examination and internal transcribed spacer (ITS)

sequence analysis indicated that the specimens represented Gymnopus (Pers.)

Roussel (Omphalotaceae, Agaricales).

Although 329 Gymnopus names have been published, only 204 are currently

accepted in the genus (http://www.indexfungorum.org/Names/Names.asp —

accessed Oct 9. 2015). China is very rich in fungal diversity, but only 17 species

of Gymnopus have been reported in the country (Li et al. 2006, Bau et al. 2007,

Wu et al. 2009, Deng et al. 2011, MeSic¢ et al. 2011, Zhang & Liu. 2011, Zhang

et al. 2012, Li et al. 2012); there are probably many more taxa to be discovered.

664 ... Deng & al.

Morphological examination of the gymnopoid collections supports

placement in Gymnopus subsect. Vestipedes, which is characterized by small

to medium-sized basidiomata, a membranaceous or thin-fleshed pileus, a

non-grooved and rootless stipe, a sterile lamella edge with well-differentiated

cheilocystidia, and a pileipellis usually with some weakly to distinctly coralloid or

diverticulate terminal elements (Antonin & Noordeloos 1997). Most members

of this subsection were formerly placed in Collybia sect. Vestipedes (Kithner

1980, Halling 1983, Jansen 1991), but they were transferred to Gymnopus

subsect. Vestipedes after Collybia sensu lato was divided into Collybia sensu

stricto, Gymnopus, and Rhodocollybia (Antonin & Noordeloos 1997). After

comparison with other species in G. subsect. Vestipedes, our Chinese taxon was

determined to represent a new species, described here as G. ramulicola.

Material & methods

Morphological studies

The description of the new species is based on specimens and field photos. Macroscopic

features were recorded in the field. Colour codes in the macroscopic description follow

Kornerup & Wanscher (1978). Microscopic features were observed from dried material

after being sectioned and mounted in 5% potassium hydroxide (KOH) or 1% Congo

Red under an Olympus BX51 light microscope (Tokyo). The notation (a—)b-c(-d) is

used to present dimensions where the range b-c represents over 90% of the measured

values, and both ‘a’ and ‘d’ are the extreme values; “n” refers to the number of measured

basidiospores, “Q” refers to the length/width ratio of basidiospores, and Q__ refers to the

mean ratio. Holotype and paratypes are deposited in the Fungal Herbarium, Guangdong

Institute of Microbiology, Guangzhou, China (GDGM).

Molecular procedures and phylogenetic analyses

Genomic DNA was extracted from dried specimens with the Sangon Fungus

Genomic DNA Extraction kit (Sangon Biotech Co., Shanghai) according to

manufacturer's instructions. The primers ITS1 and ITS4 (White et al. 1990, Gardes &

Bruns 1993) were used to amplify nuclear ribosomal internal transcribed spacer region,

containing the ITS1, 5.8S and ITS2 regions. Dideoxy sequencing was performed by

Beijing Genomic Institute (Guangzhou, China) using the ABI 3730xl DNA sequencer

(Applied Biosystems, Foster City, CA, USA). ITS sequences from 22 species taken from

GenBank were used to construct a maximum likelihood tree after these sequences

were edited and aligned by MEGA 6.0. Relative robustness of individual branches was

estimated by bootstrapping 1000 replications.

Results

Molecular results

The phylogenetic analysis contains 25 sequences, among which 22 were

downloaded from GenBank. Sequences JN714927 (Marasmius type species

Gymnopus ramulicola sp. nov. (China) ... 665

Fic. 1. Gymnopus ramulicola basidiomata.

a, b. holotype, GDGM 43884; c. GDGM 44256; d. GDGM 50060. Scale bars = 10 mm.

M. rotula (Scop.) Fr.) and JN021008 (Collybia type species C. tuberosa (Bull.)

P. Kumm.) were selected as outgroup. Maximum likelihood analysis of ITS

sequences representing Gymnopus sections Gymnopus, Vestipedes, Laevipedes,

and Androsacei placed our three Chinese sequences in a single lineage and

supported our new species as a member of G. sect. Vestipedes.

Taxonomy

Gymnopus ramulicola T.H. Li & S.F. Deng sp. nov. Figs 1, 2

MycoBank MB 815128

Differs from Gymnopus melanopus by its pale red to pinkish white pileus, shorter and

white to greyish red or dull red stipe, smaller basidiospores, and smaller cheilocystidia;

and from G. confluens by its smaller pileus, distant lamellae, shorter stipe, and its

gregarious habit on fallen twigs.

Type: China, Hainan Province: Ledong County, Jianfengling National Forest Park,

18°43’36’N 108°53’56”E, alt. 787 m asl, 29 June 2013, leg. Wang CQ, Zhou SH

(Holotype, GDGM 43884; GenBank KU057798).

Etymology: epithet ramulicola indicates that the new species is growing on fallen twigs.

Piteus 5-25 mm broad, hemispherical to convex when young, expanding to

applanate with a slightly uplifted margin, pale red (7A3, 8A3, 9A3) to pastel red

(7A4, 8A4) at disc and reddish or pinkish white (9A2, 10A2) at margin when

666 ... Deng & al.

Fic. 2. Gymnopus ramulicola. (holotype, GDGM43884).

a. basidiospores; b. basidia and basidioles; c. cheilocystidia ; d. pileipellis. Scale bars = 10 um.

young, sometimes greyish orange (6B3-4), greyish red (7B3-4) or dull red

(8B3, 8C3), usually paler when mature, becoming orange white or pale orange

(5A2-3, 6A2-3) to yellowish white or pale yellow (3A2-3, 4A2-3) when old,

dry, with inconspicuous radial stripes or grooves and fine white tomentum.

LAMELLAE adnate to sub-free, narrow, distant, with 9-12 complete lamellae

and 1-4 lamellulae of different lengths between the two complete lamellae,

concolourous with pileus to somewhat dull red or greyish red (8B4 to 9B5,

8C4-5). Stipe central, 12-23 x 2-3 mm, cylindrical, hollow, often slightly

Gymnopus ramulicola sp. nov. (China) ... 667

inflated at the base, (sub)insititious, white at apex, becoming greyish red

(7B3-4) or dull red (8B3, 8C3) downwards, dry, covered with fine white

tomentum, and with white mycelium at base. Context thin, whitish to similar

to pileus colour. ODouR and TASTE not distinctive.

BASIDIOSPORES (n = 30) (6.3-)6.6-8.4(-8.8) x (3.2-)3.5-4.5 um, Q = 1.6-2.4,

Q_, = 1.94, ellipsoid to oblong, smooth, thin walled, hyaline. Basrp1a 23-27

(-30) x 3.8-5.5 um, clavate to cylindrical, inflated at apex, thin-walled, with

(2-)4 sterigmata, clamped. BasIDIOLES 16-25 x 3.0-4.5 um, cylindrical,

clamped. CHEILOCYSTIDIA 23-27(-30) x 3-6 um, indistinct, irregularly clavate

to cylindrical, inflated at apex, hyaline, thin-walled. PLEURocysTip1A absent.

PILEIPELLIS a cutis of radially arranged cylindrical hyphae which are 4-9 um

in diam., smooth, septate, thin-walled, and weakly to distinctly diverticulate.

CLAMP CONNECTIONS abundant and present in all tissues.

HasBiTaT—gregarious on fallen twigs in tropical or subtropical broadleaf

forests.

ADDITIONAL SPECIMENS EXAMINED: CHINA, HAINAN PROVINCE: Changjiang County,

Bawangling National Forest Park, 19°07’25”N 109°05’35”E, alt. 657 m asl, 6 June 2013,

leg. Wang CQ, Zhou SH (GDGM 44256; GenBank KU321529); JIANGxXI PROVINCE:

Ganzhou City, Chongyiyang National Forest Park, 55°38’99”N 114°19’10’E, alt. 374 m

asl, 2 August 2013, leg. Zhang M, Xu J, Zhou SH (GDGM 50060; GenBank KU321530).

Discussion

Among the species with ITS sequences in the phylogenetic tree (Fic. 3),

Gymnopus ramulicola is closest to G. melanopus A.W. Wilson et al., which

differs by its dark brown to greyish orange or beige pileus, its longer (21-55

mm) brown to black stipe, its larger basidiospores (7.2-10.4 x 3.2-5.6 um),

and its much larger cheilocystidia (32-56 x 9.5-16.5 um; Wilson et al. 2004).

The second closest taxon in the tree is G. nonnullus var. attenuatus (Corner)

A.W. Wilson et al., which differs by its dark chocolate brown to grayish brown

pileus, its larger basidiospores (7.2-9.6 x 3.2-4.8 um), and its much larger

cheilocystidia (37.5-60 x 5.5-7 um; Wilson et al. 2004).

Morphologically, G. confluens (Pers.) Antonin et al., G. huijsmanii Antonin

& Noordel., G. inodorus (Pat.) Antonin & Noordel., G. moseri Antonin &

Noordel., and G. putillus (Fr.) Antonin et al. (all in G. subsect. Vestipedes;

Antonin & Noordeloos 1997) somewhat resemble G. ramulicola. However,

G. confluens has a bigger (10-65 mm), pale brown, yellow-brown or pale grey-

brown pileus, more crowded lamellae, and a longer, distinctly non-insititious

stipe (25-90 x 1.5-4 mm; Halling 1983, Antonin & Noordeloos 1997);

G. huijsmanii has incarnate to violaceous brown stipe, larger basidiospores

(6.5-11.5 x 3.5-5.5 um; Antonin & Noordeloos 1997); G. inodorus has more

668 ... Deng & al.

Gymnopus confluens KJ817065

Gymnopus tamblinganensis AY263450

80 Gymnopus cylindricus DQ450057

97 74 Gymnopus menehune DQ450043

Gymnopus neotropicus AF505769,

50 Gymnopus collybioides AF 505772

Gymnopus nonnullus var.attenuatus AY 263445.

Gymnopus melanopus AY263425

ot Sect. Vestipedes

50 Gymnopus ramulicola KU057798

1 Gymnopus ramulicola KU321529

Gymnopus ramulicola KU321530

Gymnopus trogioides AY263428

100 2 3 Gymnopus brunneigracilis AY263434

Gymnopus gibbosus DQ450020

Gymnopus luxurians DQ450024

Gymnopus fuscotramus JF303730 | Sect. Androsacei

Gymnopus fusipes AF505777 | Sect. G ymnopus

Gymnopus indoctoides AY 263424

Gymnopus bicolor AY263423

Gymnopus spongiosus DQ480113 S ect TE: ae vip e de s

Gymnopus alpinus DQ480114

Gymnopus hybridus DQ449980

Gymnopus ocior DQ449968

Collybia tuberosa JNO21008

Marasmius rotula JN714927

0.05

Fic. 3. Phylogenetic tree based on the ITS rDNA region (ITS1, 5.88 rDNA and ITS2), maximum

likelihood tree after sequences being edited and aligned by MEGA 6.0. Relative robustness of

individual branches was estimated by bootstrapping 1000 replications. Bootstrap values (>50%)

are shown above individual nodes.

crowded lamellae and strongly inflated cheilocystidia that typically have finger-

like apical excrescencies (Antonin & Noordeloos 1997); G. moseri has a brown

to red-brown stipe and larger basidiospores (8.5-11.0 x 4.0-5.0 um; Antonin

& Noordeloos 1997); and G putillus has more crowded lamellae, a brown or

greyish brown, white to greyish pruinose stipe, and no cheilocystidia (Antonin

& Noordeloos 1997);

Among the other species in Gymnopus sect. Vestipedes with clamp

connections, thin-fleshed pileus, and a non-grooved and non-insititious stipe,

G. allegreti (De Seynes) A.W. Wilson et al. is distinguished by a light brown to

pale yellowish brown pileus, a longer stipe with coarse rhizomorphs, and larger

Gymnopus ramulicola sp. nov. (China) ... 669

cheilocystidia (33-46.5 x 6.5-10 um; Wilson et al. 2004); G. brunneigracilis

(Corner) A.W. Wilson et al. has a rusty reddish brown to light brown pileus and

larger basidiospores (8-13.6 x 4.0-6.1 um; Wilson et al. 2004); G. collybioides

(Speg.) Desjardin et al. has smaller basidiospores (5.6-7.2 x 2.4-4.0 um) and

basidia (16-18 x 4-5 um; Mata & Petersen 2003); G. cylindricus J.L. Mata has

crowded lamellae and longer basidiospores (8.8-13.6 x 3.2-4.8 um; Mata et al.

2004); G. indoctus (Corner) A.W. Wilson et al. has a brownish beige or creamy

beige to white pileus, a longer, fibrillose stipe with rhizoids at the base, smaller

basidia and somewhat larger cheilocystidia (26.5-35 x 9-12 um; Wilson et al.

2004); G. johnstonii (Murrill) A.W. Wilson et al. has a dark brownish grey to

brown pileus, more crowded lamellae, smaller basidia (12-17.5 x 5.5-10.5 um),

and cheilocystidia bifid or with a few lobes (Wilson et al. 2004); G. luxurians

(Peck) Murrill has a larger, reddish brown to light brown pileus, more crowded

lamellae, a stipe with rhizoids, and somewhat diverticulate cheilocystidia

(Halling 1983); G. menehune Desjardin et al. has a reddish brown to dark

brown or brown young pileus, close to crowded lamellulae, larger basidiospores

(7.5-9.5 x 3.5-4.2 um), and lobed and diverticulate cheilocystidia (16-42 x

3.8-10.0 um; Desjardin et al. 1999); and G. virescens A.W. Wilson et al. has a

dark reddish brown to light brown or light grey pileus, crowded lamellulae,

a few coarse rhizomorphs at the base of stipe, larger basidiospores (7-10 x

3-6 um), and a lamellar edge lacking cheilocystidia (Wilson et al. 2004).

Acknowledgments

Thanks are given to Shi-Hao Zhou, Ming Zhang and Jiang Xu for their help

in collecting the specimens. We are grateful to Dr. Vladimir Antonin (Moravian

Museum, Department of Botany, Brno, Czech Republic) and Dr. Armin MeSi¢ (Ruder

BoSskovic Institute, Zagreb, Croatia) for their pre-submission reviews. This work was

supported by the National Natural Science Foundation of China (Nos. 31260011,

31570020), the Ministry of Science and Technology of the People’s Republic of China

(Nos. 2014CB460613, 2013FY111200) and the Science and Technology Planning

Project of Guangzhou (No. 201504291620324).

Literature cited

Antonin V, Noordeloos ME. 1997. A monograph of Marasmius, Collybia and related genera in

Europe. Part 2: Collybia, Gymnopus, Rhodocollybia, Crinipellis, Chaetocalathus, and additions

to Marasmiellus. Libri Botanici 17. 256 p.

Bau T, Bulakh YM, Zhuang JY, Li Y. 2007. Agarics and other macrobasidiomycetes from Ussuri

River Valley. Mycosystema 26(3): 349-368.

Deng CY, Li TH, Song B. 2011. A revised checklist of Marasmiellus for China mainland. Czech

Mycol. 63(2): 203-214.

Desjardin DE, Halling RE, Hemmes DE. 1999. Agaricales of the Hawaiian Islands. 5. The genera

Rhodocollybia and Gymnopus. Mycologia 91(1): 166-176. http://dx.doi.org/10.2307/3761206

670 ... Deng & al.

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes —

application to the identification of mycorrhizae and rusts. Molec. Ecol. 2: 113-118.

http://dx.doi.org/10.1111/j.1365-294X.1993.tb00005.x

Halling RE. 1983. The genus Collybia (Agaricales) in the northeastern United States and adjacent

Canada. Mycol. Mem. 8. 148 p.

Jansen AE. 1991. Het geslacht Collybia. Wetensch. Meded. Kon. Ned. Natuurhist. Ver. 205: 64 p.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. London, Eyre Methuen.

Kihner R. 1980. Les hyménomycetes agaricoides (Agaricales, Tricholomatales, Pluteales, Russulales).

Bull. Mens. Soc. Linn. Lyon 49(num. spéc.): 1027 p.

Li JZ, Chen SM, Dan XQ. 2006. Macrofungus resources of the Shunhuang Mountain Natural

Protection Region. Life Sci. Res. 10(3): 276-281.

Li SJ, Zhu TH, Liu GH, Zhu HM. 2012. Diversity of macrofungal community in Bifeng

Gorge: the core giant panda habitat in China. African J. Biotechnol. 11(8): 1970-1976.

http://dx.doi.org/10.5897/AJB11.3232

Mata JL, Petersen RH. 2003. Type studies of neotropical Collybia species. Mycotaxon 86: 303-316.

Mata JL, Halling RE, Petersen RH. 2004. New species and mating system reports in Gymnopus

(Agaricales) from Costa Rica. Fungal Diversity 16: 113-129.

MeSi¢é A, Tkaléec Z, Deng CY, Li TH, Ple&e B, Cetkovié H. 2011. Gymnopus fuscotramus

(Agaricales), a new species from southern China. Mycotaxon 117: 321-330.

http://dx.doi.org/10.5248/117.321

White TJ, Bruns TD, Lee SB, Taylor JW. 1990. Amplification and direct sequencing of

fungal ribosomal RNA genes for phylogenies. 315-322, in: MA Innis et al. (eds).

PCR protocols, a guide to methods and applications. Academic Press, San Diego.

http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

Wilson AW, Desjardin DE, Horak E. 2004. Agaricales of Indonesia. 5. The genus Gymnopus from

Java and Bali. Sydowia 56(1): 137-210.

Wu XL, Li TH, Song B. 2009. The ecological distribution and resources of the macrofungi in

Huaping Nature Reserve of Guangxi, China. Mycosystema 28(4): 528-534.

Zhang J, Liu PG. 2011. Wild economic macrofungi in Bijie District of Guizhou province, southwest

China. Chin. J. Ecol. 30(6): 1177-1184.

Zhang Y, Zhou TS, Ou XK, Zhou DQ. 2012. New records and distribution of macrofungi in Laojun

Mountain, Northwest Yunnan, China. Mycosystema 31(2): 196-212.

MY COTAXON

July-September 2016— Volume 131, pp. 671-678

http://dx.doi.org/10.5248/131.671

Hemithecium hainanense sp. nov. and

a checklist and key to Hemithecium species

from China

ZE-FENG JIA*, MEI-FANG LI & XIN ZHAO

College of Life Sciences, Liaocheng University, Liaocheng, 252059, China

* CORRESPONDENCE TO: Zffia2008@163.com

ABSTRACT—Hemithecium hainanense from Hainan Island, China, is described as new to

science. The lichen is characterized by erumpent lirellae with a lateral to complete thalline

margin, large transparent transversely septate ascospores, and the presence of norstictic acid.

Hemithecium chlorocarpum is reported as a new record from China. A checklist and key to

Hemithecium species from China are presented.

Key worp—Ascomycota, Graphidaceae, Ostropales, Ostropomycetidae, taxonomy

Introduction

The lichenized fungal genus Hemithecium Trevis. (Graphidaceae, Ostopales)

has been recently revised (Staiger 2002; Staiger et al. 2006; Licking et al. 2008;

Rivas Plata et al. 2012, 2013; Lumbsch et al. 2014; Miadlikowska et al. 2014).

The genus is characterized by its crustose thalli, lirelliform apothecia, well-

developed convergent labia, concealed discs, non-carbonized exciples, simple

paraphyses, and hyaline transversely septate or muriform ascospores.

Hemithecium contains about 50 species worldwide (Staiger 2002, Upreti

et al. 2011, Sharma et al. 2012). Miao et al. (2007) reported three species of

Hemithecium (H. chapadanum, H. chlorocarpoides, and H. chrysenteron)

as new records from China, which Liicking et al. (2008) since placed in the

invalid genus “Pallidogramme”. During the compilation of the Flora Lichenum

Sinicorum, five Hemithecium species were recognized in China (Jia et al. 2011,

Jia & Wei 2016). Here, we describe a new species, Hemithecium hainanense, and

add a new Chinese record, H. chlorocarpum.

672 ... Jia, Li & Zhao

Materials & methods

The specimens are deposited in the Herbarium Mycologicum Academiae Sinicae

— Lichenes, Beijing, China (HMAS-L) and the Lichen Herbarium of the College of

Life Sciences, Liaocheng University, Liaocheng, China (LCU-L). Morphological and

anatomical features were examined using an Olympus SZX12 dissecting microscope and

Olympus BX51 & Nikon Eclipse-55i light microscopes. Measurements and illustrations

were taken from the hand cut cross-sections of fruit bodies in tap water. Amyloidity of

the ascospores was tested using Lugol's solution. The thallus surface and thin thallus

sections were spot tested with K (KOH, 20%), C (saturated solution of NaOCl), and

P (ethanol solution of p-phenylenediamine, 5%) reagents. The lichen substances were

detected and identified by thin-layer chromatography (TLC; Culberson & Kristinsson

1970, Culberson 1972, White & James 1985).

Taxonomy

Hemithecium hainanense Z.F. Jia, sp. nov. PL.1

FUNGAL NAME FN570071

Differs from Hemithecium implicatum by its longer ascospores and presence of norstictic

acid.

Type: China. Hainan Province, Changjiang County, Mt. Bawangling, Yajia, 19°14’N

109°26’E, alt. 800 m, on bark, 10/V/2008, J. Li HN081472 (Holotype, HMAS-L 127460).

ErymMo ocy: The specific epithet hainanense is derived from the type locality, Hainan,

and -ense, Latin, place of origin.

THALLUS corticolous, crustose, pale to grayish white, thin, slightly smooth,

tightly attached to the substratum, lacking isidia and soredia.

APOTHECIA lirelliform, numerous, elongate, simple, rarely branched,

erumpent, not striate, 1-4 mm long, 0.1—0.2 mm wide, covered by lateral

to complete thalline margin; pisc concealed, with slit-like appearance;

PROPER EXCIPLE conspicuous, convergent, uncarbonized, yellowish to

brown; HYMENIUM clear, 110—180 um high, I-, with brownish epihymenium;

HYPOTHECIUM brownish, 5-15 um high. ParapuHyses simple, 1-2 um

wide. Asci clavate, 90-130 x 15-20 um, 6—8-spored. Ascosporss hyaline,

surrounded by a thin halo, narrowly ellipsoidal, transversely septate, with

17—20 locules, 50-100 um long and 6.5—10 um wide, I+ violet.

CHEMISTRY: K+ red, C—, P+ yellow; norstictic acid present (TLC).

SUBSTRATE: on bark.

ADDITIONAL SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Changjiang County,

Mt. Bawangling, alt. 800 m, 10/V/2008, J. Li HN081467 (HMAS-L 127462), HN081470

(HMAS-L 127461); Qiongzhong County, Mt. Limushan, alt. 650 m, 25/IX/2009, B. Gao

HN080910 (HMAS-L 127464), J. LiHN081236 (HMAS-L 127463); Ledong County, Mt.

Jianfengling, alt. 900 m, M. Liu HN09414 (HMAS-L 115510).

ECOLOGY & DISTRIBUTION: Hemithecium hainanse is known from altitudes of

650-900 m in a tropical rainforest on Hainan Island in southern China where

Hemithecium hainanense sp. nov. (China) ... 673

PLATE 1. Hemithecium hainanense (holotype, HMAS-L 127460). A. Thallus on bark; B. Cross

section of apothecium; C—E. Asci containing ascospores (showing the process of ascospore

development and maturation); F Mature ascospores. Scale bars: A = 1 mm; B = 100 um;

C-F = 50 um.

it grows on smooth bark in moist and humid places. Associated lichen species

include Diorygma hieroglyphicum (Pers.) Staiger & Kalb, Graphis hossei Vain.,

G. japonica (Mill. Arg.) A.W. Archer & Liicking, “Pallidogramme chrysenteron”

(Mont.) Staiger et al., and other members of the Graphidaceae.

REMARKS: Hemithecium hainanense is characterized by simple and sparsely

branched erumpent lirellae with a lateral to complete thalline margin, a

concealed disc, an uncarbonized exciple, a clear hymenium, transversely septate

ascospores of 50—100 x 6.5—10 um, and the presence of norstictic acid. The new

species resembles Hemithecium implicatum but is distinguished from the latter

by the larger transversely septate ascospores and the presence of norstictic acid.

674 ... Jia, Li & Zhao

Hemithecium balbisii (Fée) Trevis., Spighe e Paglie: 13 (1853).

THALLUS corticolous, greenish-gray, upper surface waxy. APOTHECIA

lirelliform, 2-8 mm long, 0.3-0.4 mm wide, sometimes branched, covered partly

by the cream-colored, indistinctly crenate thalline margin. Discs concealed.

PROPER EXCIPLE convergent, pale to brown. HyMENIvM clear, 80-90 um high.

Ascosporss 8/ascus, hyaline, elongate, muriform, 10-12 x 2-4-locular, 28-50

um long and 10-15um wide, I+ violet, lacking a surrounding halo.

CHEMISTRY: K—, C—, P-; no substances present (TLC).

SUBSTRATE: on bark.

SPECIMENS EXAMINED: CHINA. FUJIAN PROVINCE, Wuyishan City, Mt. Wuyishan,

alt. 540 m, 27/V/2007, J. Li FJ502 (LCU-L). HAINAN PROVINCE, Baoting County, Mt.

Qixianling, alt. 200 m, 23/VII/2009, J. Li HN240-1 (HMAS-L 125725); Ledong County,

Mt. Nanlimu, alt. 180 m, 22/VII/2009, M. Liu HN09357 (HMAS-L 115556). YUNNAN

PROVINCE, Honghe City, Mt. Daweishan, alt. 900 m, 23/IX/2011, Z.F Jia 11-455

(LCU-L); 1250 m, 23/IX/2011, Z.F. Jia 11-508-3 (LCU-L).

REMARKS: The species is similar in appearance to “Pallidogramme chlorocarpoides,”

which differs by its larger, brown ascospores and its production of stictic acid.

Hemithecium balbisii was previously reported from an unstated locality in

China as Graphis balbisii Fée (Krempelhuber 1868; Wei 1991). A pantropical

species, H. balbisii also occurs in South America (Staiger 2002).

Hemithecium canlaonense (Vain.) A.W. Archer, Syst. Biodiv. 5: 17 (2007).

THALLUS corticolous, gray to greenish-gray, upper surface waxy. APOTHECIA

lirelliform, 1-3 mm long, 0.2-0.3 mm wide, sometimes branched, covered partly

by the indistinctly crenate thalline margin. Discs concealed. PROPER EXCIPLE

convergent, pale to brown. HyMENIuM clear, 80-110 um high. AscosporEs

8/ascus, hyaline, ellipsoidal, transversely septate, 4-locular, 15-25 um long and

8-14 um wide, I-, lacking a surrounding halo.

CHEMISTRY: K—, C—, P-; no substances present (TLC).

SUBSTRATE: on bark.

SPECIMENS EXAMINED: CHINA. GUIZHOU PROVINCE, Tongren City, Mt. Fanjingshan,

alt. 133 m, on bark, 16/IX/1963, J.C. Wei 0762 (HMAS-L 047793), 0763 (HMAS-L

047790).

REMARKS: The species is similar to H. oshioi, but differs by its smaller ascospores

and the absence of stictic acid. Hemithecium canlaonense was previously

reported in China as Fissurina canlaonensis (Vain.) Staiger (Meng et al. 2011).

This pantropical species also occurs in Indonesia and the Philippines (Archer

2007) and in India (Patwardhan & Kulkarni 1976; Sharma et al. 2012) where it

was recorded as Fissurina canlaonensis.

Hemithecium hainanense sp. nov. (China) ... 675

Hemithecium chlorocarpum (Fée) Trevis., Spighe e Paglie : 12 (1853).

THALLuUS corticolous, pale gray to greenish-gray, surface smooth.

APOTHECIA lirelliform, 1-4 mm long, 0.2-0.4 mm wide, sometimes branched,

covered partly by the distinctly crenate, thalline margin. Discs concealed.

PROPER EXCIPLE convergent, pale to brown. HyMENIUM clear, 150-180 um

high. Ascosporss 1/ascus, hyaline, ellipsoidal, muriform, 20-30 x 5-8-locular,

85-125 um long and 18-27.5 um wide, I+ violet, lacking a surrounding halo.

CHEMISTRY: K—, C—, P-; no substances present (TLC).

SUBSTRATE: on bark.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Wuzhishan City, Mt. Wuzhishan,

alt. 750 m, on bark, 28/V1/2001, J.B. Chen 20657 (HMAS-L 076854)

REMARKS: The species resembles Hemithecium balbisii, but differs by its larger

ascospores and asci containing one ascospore. Hemithecium chlorocarpum is

here reported as new to China. As a pantropical element, it also occurs in South

America (Staiger 2002).

Hemithecium duomurisporum Z.F. Jia, Mycosystema 30: 871 (2011).

THALLUS corticolous, grayish to olive-green, smooth. APOTHECIA

lirelliform, 2-8 mm long and 0.3-0.5 mm wide, rarely branched, covered partly

by the slightly crenate thalline margin. Discs concealed. PROPER EXCIPLE

convergent, yellow-brown. HyMENtIuM clear, 150-180 um high. AscosporEs

1/ascus, hyaline, lacking a surrounding halo, elongate, muriform, with 20-22 x

3-9-locular with the traverse septa doubled, 100-140 um long and 20-29 um

wide, I+ violet.

CHEMISTRY: K+ yellow, C—, P+ orange; stictic acid present (TLC).

SUBSTRATE: on bark.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Wuzhishan City, Mt. Wuzhishan,

alt. 640 m, 20/VII/2009, M. Liu HN09318 (HMAS-L 115536); alt. 680 m, 28/IX/2008,

Z.E, Jia HN0080706 (HMAS-L 127486). Qiongzhong County, Mt. Limushan, 750 m, 25/

X1/2008, J, Li HN081229 (HMAS-L 126329).

REMARKS: This species is characterized by the large muriform ascospores and

presence of stictic acid. It is similar to H. chlorocarpum, which differs by its

smaller ascospores and its lack of lichen substances (see the description above).

Hemithecium duomurisporum was previously recorded from Hainan, China

(Jia et al. 2011).

Hemithecium implicatum (Fée) Staiger, Biblioth. Lichenol. 85: 287 (2002).

THALLUS corticolous, greenish, slightly smooth. APOTHECIA lirelliform,

1-5 mm long and 0.1-0.2 mm wide, often branched, covered partly by the

676 ... Jia, Li & Zhao

indistinctly crenate thalline margin. Discs concealed, with slit-like appearance.

PROPER EXCIPLE convergent, yellowish to brown. Hymenium clear, 100-150

um high. Ascospores 2-8/ascus, hyaline, lacking a surrounding halo, elongate,

transversely septate with 15-23 locules, 35-80 um long and 5.5-9 um wide,

I+ violet.

CHEMISTRY: K—, C—, P-; no substances present (TLC).

SUBSTRATE: on bark.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Ledong County, Mt. Jianfengling,

900-950 m, 26-28/VII/2009, J. Li HN09127 (HMAS-L 115454), HN09128 (HMAS-L

115437), HN09218 (HMAS-L 115502); Changjiang County, Mt. Bawangling, 880

m, 25/X/2008, J. Li HN081456 (HMAS-L 117055), HN081477 (HMAS-L 126330),

HN081478 (HMAS-L 126298); Qiongzhong County, Mt. Limushan, 650 m, 25/IX/2008,

J. Li HN081237 (HMAS-L 117096), HN081465 (HMAS-L 117097); Wuzhishan City, Mt.

Wuzhishan, 680 m, 20/VII/2009, J. Li HN09064 (HMAS-L 115475), HN204 (HMAS-L

117067).

REMARKS: This species is similar in appearance to H. oshioi, which is

distinguished by its shorter ascospores and its production of lichen substances

(see the description below). Hemithecium implicatum also occurs in Mexico

(Wirth & Hale 1963, as Graphis implicata) and Brazil (Staiger 2002).

Hemithecium oshioi (M. Nakan.) M. Nakan. & Kashiw., Bull. Natl. Sci. Mus. Tokyo,

B 29(2): 88 (2003).

THALLUS corticolous, grayish, rough. APOTHECIA lirelliform, 0.5-3 mm

long and 0.2-0.4 mm wide, rarely branched, covered partly by the indistinctly

crenate thalline margin. Discs concealed. Proper exciple convergent, yellowish

to brown. HyMENIvuM clear, 90-120 um high. Ascospores 2-6/ascus, hyaline,

lacking a surrounding halo, elongate, with 5-9 transverse septa, 25-40 um long

and 5.5-8 um wide, I+ violet.

CHEMISTRY: K+ red, C—, P+ yellow; norstictic acid present (TLC).

SUBSTRATE: on bark.

SPECIMEN EXAMINED: CHINA. FUJIAN PROVINCE, Jianou City, Fangdao Town,

Wanmulin, 310 m, 3/VI/2007, Q.F. Meng FJ925 (LCU-L). Hainan PROVINCE,

Wuzhishan City, Mt. Wuzhishan, 700 m, 21/VII/2009, J. Li HN09092 (HMAS-L

115471); 670 m, 28/1X/2008, T. Zhang HN080346 (HMAS-L 125726). Ledong County,

Mt. Jianfengling, 850 m, 29/VII/2009, Z.F. Jia HNJ63 (HMAS-L 128787), HNJ64

(HMAS-L 128786).

REMARKS: This species is similar to H. laubertianum, which differs by its thick-

walled spores and its lack of lichen substances (Staiger 2002). Hemithecium

oshioi was reported from Zhejiang, China (as Graphis oshioi M. Nakan.: Wu &

Qian 1989, Wei 1991) and Hainan, China (Jia et al. 2011, Wei et al. 2013). It is

also recorded from Japan (Nakanishi 1966, as Graphis oshioi).

Hemithecium hainanense sp. nov. (China) ... 677

Key to species of Hemithecium from China

LAScospores-withy transverse Septal §5..iy sk bsg ences’ se boa Ee bel Eh pow Fn pocbe a peces pest z

TRASCOSpObes Turion ities S20. oF Noe oes eh ee Fa he Same ea rtR NUR cee gBOR veo 5

2. Ascospores 4-locular, 15-25 x 8-14 um ...... 0. eee eee H. canlaonense

2. Ascospotes SAtl@eilar, a2 SUNOS “sn Wee ha ee Battal eee Rades ore Ratan SR ade: cette: whether 3

3. No substances present; ascospores 15-23-locular, 35-80 x 5.5-9 um_ H. implicatum

3. NOBSTICIEC CIC, DECSEM es Surtd. aild ye tail dee peat oen al gh oak gooeehd rape ed eh peed ea See eke 4

4, Ascospores 17-20-locular, 50-100 x 6.5-10 Um ....... 2... eee eee H. hainanense

4, Ascospores 6-10-locular, 25-40 x 5.5-8 UM ... 00... eee eee eee H. oshioi

5. Stictic acid present; ascospores 100-140 x 20-29 Um.......... H. duomurisporum

3 Nowsubstancescsresemt gee te. Seger. te yo ee Re Ree re re Ur Re Oe 6

Gi ASCOSP Ones ea SRLS WS BLIMae Bet np aot Bao oh Banat he Eanes cae tetas H. balbisii

6: ASCOSPOTES. B54 125 418-274 Sis se abe pata poka by pasa ram post « H. chlorocarpum

Acknowledgements

This study was supported by the National Natural Science Foundation of China

(31270066, 31093440, 31493010 & 31493011) and Shandong Excellent Youth Scientist

Foundation Project (BS2011SW028). The authors are grateful to Dr. Alan W. Archer

and Dr. Hua-Jie Liu for reading and improving the manuscript and for acting as

presubmission reviewers.

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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, Kristinsson H. 1970. A standardized method for the identification of lichen products.

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MY COTAXON

July-September 2016— Volume 131, pp. 679-685

http://dx.doi.org/10.5248/131.679

First records of Faurelina in the Neotropics

ROGER FAGNER RIBEIRO MELO '*, ANDREW N. MILLER ”

& LEONOR CosTA Mata !

' Universidade Federal de Pernambuco, Departamento de Micologia,

Av. da Engenharia, s/n, 50740-600, Recife, Brazil

? University of Illinois at Urbana-Champaign, Illinois Natural History Survey,

1816 S. Oak St., Champaign, IL 61820, USA

" CORRESPONDENCE TO: rogerfrmelo@gmail.com

ABSTRACT—Two species of Faurelina, F. fimigena and EF. hispanica, are recorded fruiting

on herbivore dung collected in the Caatinga biome of the semi-arid region of Brazil. These

represent the first records of this genus in the Neotropics. Descriptions, a photographic plate

and a comparative table are provided, along with an identification key to the four species of

Faurelina.

Key worps—coprophilous fungi, Didymellaceae, non-ostiolate ascomycetes, Pleosporales

Introduction

Named after the French mycologist Louis Faurel (1907-73), Faurelina

Locq.-Lin. was first described by Locquin-Linard (1975) as a new genus of

cleistothecial ascomycetes. The genus has since had a problematic taxonomical

history. At first Faurelina was tentatively placed in Chadefaudiellaceae, but

this position was doubtful. Chadefaudiellaceae was validated by Benny &

Kimbrough (1980), based on the suggestion by Faurel & Schotter, to include

species with non-ostiolate immersed ascomata with a pseudoparenchymatous

peridium and aerial thick-walled ‘capillitium, enclosing catenulate, evanescent

asci with non-dextrinoid ascospores devoid of germ pores (Cannon & Kirk

2007).

After examining the type species Faurelina fimigena [as F. fimigenes],

Parguey-Leduc & Locquin-Linard (1976) stated that it belonged to the

680 ... Melo, Miller & Maia

ascolocular ascomycetes and concluded that it should be placed in the class

formerly known as Loculoascomycetes. Two years later, von Arx (1978)

transferred Faurelina to Microascaceae (Microascales), based mainly on the

presence of dextrinoid ascospores without germ pores. Placement of Faurelina

in Microascales was due in part to the work of Tang et al. (2007), who sequenced

the nucLSU, nucSSU, and RPB2 gene regions from a single strain labelled as

Faurelina indica (CBS 126.78), giving results identical to those of Ceratocystis

fimbriata, the type species of Ceratocystis (Microscales). Réblova et al. (2011)

reevaluated Chadefaudiellaceae based on the suggested afhnity of Faurelina

indica Arx et al. with Microascales; they believed that the material studied by

Tang et al. (2007) was a different fungus and that the LSU sequence analysis

suggested a relationship with Didymellaceae (Pleosporales, Dothideomycetes),

thus corroborating the original hypothesis proposed by Parguey-Leduc &

Locquin-Linard (1976), that Faurelina was related to fungi with an ascolocular

development.

In Brazil, recent efforts to compile data on fungal diversity, distribution, and

biogeography of fungi, include the discovery and delimitation of taxa in each

biome and their substrate/climate relationship (Braga-Neto et al. 2013). This

work contributes to these efforts in the semi-arid regions of South America by

recording the discovery of an unreported genus in the Neotropics, represented

by two species fruiting on herbivore dung in the Brazilian Caatinga biome.

Material & methods

Dung samples were collected from farms close to the Instituto Agronémico de

Pernambuco (IPA) in Serra Talhada (7°54’59”S 38°17’00”W), in the semi-arid region

of Pernambuco, Brazil. Fresh samples of goat, cattle, and horse dung were collected

in clean plastic bags, taken to the laboratory, gently air dried when necessary, and

incubated in moist chambers at room temperature (28 + 2 °C) for at least 60 days under

alternating natural light and dark periods. The specimen habit was observed directly

from substrata under a Leica EZ4 stereomicroscope, and cleistothecia in different

stages of maturation were mounted in tap water for measurements, description and

identification under an Olympus BX51 compound microscope. High quality images

were captured with a Qimaging QColor 3 digital camera mounted on an Olympus BX51

compound microscope using differential interference or phase contrast microscopy.

Specimens were identified based on descriptions provided by Locquin-Linard

(1975), Valldosera et al. (1987), and von Arx et al. (1988). Due to lack of material, no

attempt was made to culture these specimens. Permanent slides were prepared with

polyvinyl-lacto-glycerol (PVLG) and deposited at Herbarium Padre Camille Torrend,

Departamento de Micologia, Universidade Federal de Pernambuco, Recife, Brazil

(URM). Additional information regarding accessioned records, as well as photographic

data can be accessed at the “INCT - Herbario Virtual da Flora e dos Fungos” database

website (http://inct.florabrasil.net/).

First record of Faurelina (Brazil) ... 681

Ficure 1. Faurelina fimigena (URM86671): A. cleistothecium in mountant, releasing mature

ascospores; B, C. ascomatal wall; D. mature ascospores. Faurelina hispanica (URM86672):

E. cleistothecium in mountant, releasing mature ascospores; F. ascomatal wall; G. mature

ascospores. Scale bars: A = 50 um; B, C = 25 um; D = 7.5 um; E = 30 um; KF G=5 um.

682 ... Melo, Miller & Maia

Taxonomy

Faurelina fimigena Locq.-Lin., Rev. Mycol. (Paris) 39: 127 (1975) Fic. 1A-D

CLEISTOTHECIA immersed to superficial, solitary, globose to subglobose,

dark red, 190-240 um diam., with a roughened wall, glabrous. ASCOMATAL

WALL composed of inflated, globose to slightly angular stromatic cells (textura

globulosa), thick-walled, rusty red to copper in color, 5-7.5 um diam.

INTERASCAL ELEMENTS not observed in the examined material. Asci 8-spored,

globose to subglobose, often slightly clavate, 10-15.5 x 8-12.5 um, occasionally

with a short stipe, forming short chains during ascospore maturation,

evanescent, irregularly biseriate. AscosporREs variable in morphology, ellipsoid,

rhomboid or angulated, with abrupt ends, hyaline to slightly pale brown when

mature, one-celled, with longitudinal striae and furrows, thick-walled, 7.5-10

x 4.5-5.5 um.

MATERIAL EXAMINED — BRAZIL. PERNAMBUCO, Instituto Agronémico de Pernambuco

(IPA), Serra Talhada, on cattle dung, 28.VII.2012, R-ER Melo (URM86671).

Hasirat: Recorded on cattle and goat dung.

DISTRIBUTION: Asia (India, Japan), Europe (France), South America

(Brazil). This is the first record from South America.

Notes: Faurelina fimigena is the type species of the genus, described by

Locquin-Linard from material obtained in Paris. Although the records

are scarce making it difficult to delimit the species, it differs from the other

Faurelina species by ascospore shape and size. The ascospores in the Brazilian

material are longer than those cited in the original description but are similar

to the specimens analyzed by von Arx et al. (1988) obtained from subcultures

of the type material. Faurelina fimigena resembles F. elongata (Udagawa &

Furuya) Furuya [= Leuconeurospora elongata Udagawa & Furuya], and there

has been some confusion in the delimitation of these species. However,

E fimigena differs from F. elongata in ascospore length.

Faurelina hispanica Valldos. & Guarro, Mycotaxon 30: 5 (1987) Fic. 1E-G

CLEISTOTHECIA immersed to superficial, solitary, globose, dark red,

195-210 um diam., glabrous. ASCOMATAL WALL composed of inflated, globose

to slightly angled stromatic cells (textura globulosa), thick-walled, rusty red

to copper in color, 5-10 um diam. INTERASCAL ELEMENTS not observed

in the examined material. Ascr 8-spored, clavate to cylindrical-clavate,

11-12.5 x 5-9.5 um, occasionally with a short stipe, forming short chains

during ascospore maturation, evanescent, irregularly biseriate. ASCOSPORES

ellipsoidal to oblong, with rounded ends, hyaline to slightly pale brown when

TABLE 1. Synopsis of diagnostic characters in Faurelina

First record of Faurelina (Brazil) ... 683

SPECIES | CLEISTOTHECIA PERIDIUM ASCOSPORES CONIDIA

E elongata Vertically Cephalothecoid; Fusiform to ellipsoidal, Absent or

: elongated, cells thick-walled, 5-7 x 3.5-4.5 um not observed

300-360 um angular

diam.

E fimigena Globose to Non-cephalothecoid; Ellipsoidal, rhomboidal = Absent or

: subglobose, cells globose inflated, or angular, with abrupt not observed

wall roughened; thick-walled, rusty red ends, longitudinally

190-240 um to copper in color, striate and furrowed;

diam. 5-7.5 um diam. thick-walled,

7.5-10 x 4.5-5.5 um.

E hispanica : Globose, Non-cephalothecoid; Ellipsoidal to oblong, Absent or

195-210 um cells globose inflated, with rounded ends, not observed

diam. thick-walled, rusty red longitudinally striate,

to copper in color, 5-6 x 2.5-3 um

5-10 um diam.

E indica Hemispherical Non-cephalothecoid; Fusiform-navicular Arthroconidia,

or pustulate, cells thick-walled, or rhomboidal, with 1-2-celled,

rounded above, in vertical rows, some furrows, finely hyaline, with

170-300 x elongate, striated by irregular, truncate ends

180-250 tum 5-7 um diam. usually longitudinal

thickenings,

6-8 x 4-5.5 um

mature, one-celled, with longitudinal striations, 5-6 x 2.5-3 um in frontal view,

2-2.5 um wide in side view.

MATERIAL EXAMINED:— BRAZIL. PERNAMBUCO: Instituto Agrondémico de Pernambuco

(IPA), Serra Talhada, in horse dung, 14.1I.2013, R.ER Melo s.n. (URM86672).

Hasirat: Recorded on goat dung.

DISTRIBUTION: Europe (Spain), South America (Brazil). This is the first

record from South America.

Notes: Faurelina hispanica was described by Valldosera et al. (1987) and

distinguished from F fimigena by its smaller ascospores with rounded ends.

Key to species of Faurelina

1. Peridium cephalothecoid; ascomata vertically elongate ................ F. elongata

Peridium not cephalothecoid; ascomata globose to subglobose ................... 2

2. Conidial state with cylindrical, 0-1-septate arthroconidia;

ascospores fusiform-—navicular

Conidial state absent or otherwise; ascospores ellipsoidal ....................004. 3

684 ... Melo, Miller & Maia

3. Ascospores 7.5-10 x 4.5-5.5 um, irregular in shape,

abruptly.angular, with tapeted ends: oho eey cole ed bleed oie bp swe E fimigena

Ascospores 5-6 x 2.5-3 um, regular in shape,

phaseoliform to ellipsoidal, with rounded ends .................. E hispanica

Discussion

This study is the first to report Faurelina from the Neotropics, expanding

its biogeographical distribution. Both species recorded in Brazil are

morphologically very similar to the original European descriptions. The other

two Faurelina species (not yet known from Brazil) were described from Asia:

FE. elongata from Kagoshima (southwestern Kyushu, Japan) and F. indica from

Delhi (India), both fruiting on herbivore dung.

It is noteworthy that despite the relatively small number of records of this

genus worldwide, two Faurelina species were recorded in a semi-arid biome.

Conservation issues regarding Brazilian semi-arid diversity present a great

challenge to taxonomists for a number of reasons, the most significant of

which is that with approximately 98% of its territory outside conservation

areas, the semi-arid is the least protected natural area in Brazil and suffers

from environmental degradation caused by unsustainable use of its natural

resources (Leal et al. 2003). Conservation is a major concern of mycologists

in the 21“ century, who are aware of the significant decline of natural habitats

(Moore et al. 2001). The biological diversity in the Caatinga biome is high in

relation to what was previously believed. The lack of knowledge about fungal

diversity, especially coprophilous fungi, is an important factor that complicates

conservation strategies. The focused study of herbivore dung mycobiota can

improve the knowledge of the biogeography and diversity of many unreported

species.

Acknowledgments

The authors thank José Luiz Bezerra (Universidade Federal do Recéncavo da Bahia,

Brazil) and Huzefa A. Raja (University of North Carolina at Greensboro, U.S.A.) for

presubmission review.

Literature cited

Benny GL, Kimbrough JW. 1980. A synopsis of the orders and families of Plectomycetes with keys

to genera. Mycotaxon 12(1): 1-91.

Braga-Neto R, de Giovanni R., Pezzini FF, Canhos DAL, Marino A, Souza S, Maia LC. 2013. Spatial

data for fungal specimens: retrospective georeferencing and practical recommendations for

mycologists. Mycotaxon 125: 289-301. http://dx.doi.org/10.5248/125.289

Cannon PF, Kirk PM. 2007. Fungal families of the world. CABI, Wallingford.

Leal IR, Tabarelli M, Silva JMC. 2003. Ecologia e conservacao da Caatinga. Editora Universitaria

da UFPE, Recife.

First record of Faurelina (Brazil) ... 685

Locquin-Linard M. 1975. Faurelina, nouveau genre d’'Ascomycetes (Chadefaudiellaceae?). Revue de

Mycologie 39(2): 125-129.

Moore D, Nauta MM, Evans SE, Rotheroe M. 2009. Fungal conservation: issues and solutions.

Cambridge University Press, Cambridge. http://dx.doi.org/10.1017/cbo9780511565168

Parguey-Leduc A, Locquin-Linard M. 1976. Lontogénie et la structure des périthéces de Faurelina

fimigenes Locquin-Linard. Revue de Mycologie 40(2): 161-175.

Réblova M, Gams W, Seifert K. 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

Tang AMC, Jeewon R, Hyde KD. 2007. Phylogenetic utility of protein (RPB2, beta-tubulin) and

ribosomal (LSU, SSU) gene sequences in the systematics of Sordariomycetes (Ascomycota,

Fungi). Antonie van Leeuwenhoek 91: 327-349. http://dx.doi.org/10.1007/s10482-006-9120-8

Valldosera M, Guarro J, Figueras MJ. 1987. Coprophilous fungi from Spain VII. Faurelina hispanica

sp. nov. Mycotaxon 30: 5-7.

von Arx JA. 1978. Notes on Microascaceae with the description of two species. Persoonia 10: 23-31.

von Arx JA, Figueras MJ, Guarro J. 1988. Sordariaceous ascomycetes without ascospore ejaculation.

Beih Nova Hedwigia 94. 104 p.

MY COTAXON

July-September 2016— Volume 131, pp. 687-691

http://dx.doi.org/10.5248/131.687

Anaverticicladus uncinatus gen. & sp. nov.

from decaying leaves from Brazil

PHELIPE M.O. Costa !, MARCELA A. BARBOSA ”,

ELAINE MALosso ! & RAFAEL E CASTANEDA-RUIZ ?

"Centro de Biociéncias, Depto de Micologia/Laboratorio de Hifomicetos de Folhedo &

? Programa de Pés-Graduagao em Biologia de Fungos,

Universidade Federal de Pernambuco, Avenida da Engenharia, s/n,

Cidade Universitaria, 50740-600, Recife, PE, Brazil

* Instituto de Investigaciones Fundamentales en Agricultura Alejandro de Humboldt’ (INIFAT),

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

* CORRESPONDENCE TO: elaine.malosso@ufpe.br

ABSTRACT—Anaverticicladus uncinatus, collected from decaying leaves in Brazil, is described

and illustrated as a new genus and species. It is distinguished by conidiophores that are

differentiated, unbranched, erect or prostrate, septate, and pale brown, conidiogenous

cells that are monoblastic, terminal, curved to uncinate near the conidiogenous locus, and

cylindrical or subulate, and conidia that are solitary cylindrical-fusiform, uncinate, smooth

at the base but coarsely verruculose elsewhere, 5-17-euseptate, and golden brown. Conidial

secession is schizolytic.

Key worps—rainforest, neotropic, taxonomy

Introduction

The diversity of conidial fungi in the Brazilian Atlantic Forest has received

little attention (Flora do Brasil 2016). During a survey of hyphomycetes

associated with decaying plant materials in Pernambuco State, a conspicuous

fungus was collected that shows remarkable differences from all previously

described genera (Seifert et al. 2011) and is described here as new.

Materials & methods

Samples of submerged litter were placed in plastic bags for transport to the

laboratory, where they were treated according to Castafieda-Ruiz et al. (2016). Attempts

688 ... Costa & al.

to obtain this species in pure culture were unsuccessful. Mounts were prepared in 90%

lactic acid. Anatomical features were measured at a 1000x magnification under a Nikon

Eclipse Ni-U microscope using bright field optics and photographed using DIC optics

with a Nikon DS-Fi2 digital camera. The holotype was deposited in the Herbarium of

Universidade Federal de Pernambuco, Recife, Brazil (URM).

Taxonomy

Anaverticicladus P.M.O. Costa, Malosso & R.E. Castafieda, gen. nov.

MycoBank MB 816930

Differs from Verticicladus by its unbranched conidiophores, uncinate or curved

conidiogenous cells, and schizolytic conidial secession.

TYPE SPECIES: Anaverticicladus uncinatus P.M.O. Costa et al.

ETyMo_oey: Greek, ana-, meaning upwards; Latin verticicladus referring to the

hyphomycete genus Verticicladus.

Asexual fungi. ConrpropHorss differentiated, erect or prostrate, unbranched,

septate, pale brown. CONIDIOGENOUS CELLS monoblastic, curved to uncinate,

integrated, determinate. Conidial secession schizolytic. Conrp1A solitary,

acrogenous, cylindrical-fusiform, euseptate, pale brown, subconical truncate

at the base.

Anaverticicladus uncinatus P.M.O. Costa, Malosso & R.F. Castafieda, sp. nov.

FIGs 1-2

MycoBAank MB 816931

Differs from all Verticicladus spp. by its unbranched slender conidiophores, its

uncinate or slightly curved conidiogenous loci, and its euseptate conidia that secede

schizolytically.

Type: Brazil, Pernambuco State: Bonito, Mata da Chuva, 8°30’S 35°42’W, on decaying

leaves of unidentified plants, 3.III.2016, coll. RO. Costa (Holotype: URM 89939).

EryMo_oey: Latin, uncinatus, meaning hook-shaped.

CoLoniEs on the natural substratum effuse, hairy, yellowish-brown. Mycelium

mostly superficial, composed of septate, anastomosing or slightly branched,

2-3 um diam., smooth, pale brown hyphae. ConipiopHor:s differentiated,

mononematous, unbranched, erect or prostrate, bent or flexuous, slender, pale

brown or brown, subhyaline or very pale brown toward the apex, smooth,

0-3-septate, 40-70 x 3-4 um. CONIDIOGENOUS CELLS monoblastic, integrated,

terminal, determinate, subulate, curved to uncinate near and just before the

conidiogenous loci, 15-20 x 3 um, pale brown. Conidial secession schizolytic.

Conip1a solitary, acrogenous, 5-17-euseptate, 45-140 x 5-9 um, cylindrical-

fusiform, uncinate or curved at the conical-truncate, smooth-walled basal cell,

7-12 x 2-3 um, remaining wall verruculose toward the attenuated, narrowly

obtuse apex, golden brown, dry.

Anaverticicladus uncinatus gen. & sp. nov. (Brazil) ... 689

Fic. 1. Anaverticicladus uncinatus (ex holotype, URM 89939).

A, B. Conidiophores, conidiogenous cells, and conidium.

C, D. Conidiogenous cells and conidia section. E-I. Conidia.

690 ... Costa & al.

Fic. 2. Anaverticicladus uncinatus (ex holotype, URM 89939).

A-E. Conidia. F Conidiophore, conidiogenous cells, and conidium.

G-I. Conidiogenous cells and conidia.

Anaverticicladus uncinatus gen. & sp. nov. (Brazil) ... 691

ComMENTs: Anaverticicladus is superficially similar to Mycofalcella Marvanova

et al., Paraarthrocladium Matsush., and Verticicladus Matsush., all of which

have short differentiated conidiophores and a blastic conidial ontogeny but lack

uncinate or curved conidiogenous cells near or just before the conidiogenous

loci (Seifert et al 2011). Mycofalcella is distinguished by reduced unbranched

to sparingly branched conidiophores and hyaline scoleco- or phragmoconidia

with a basal appendage (Marvanova et al. 1993). Paraarthrocladium has

unbranched or branched conidiophores, the conidiogenous cells are integrated

and determinate or indeterminate with sympodial extensions, and all structures

including the conidia are hyaline; and Verticicladus is characterized by almost

verticillate branched conidiogenous cells and rhexolytic conidial secession

(Matsushima 1993).

Acknowledgments

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

S. Monteiro for their critical review of the manuscript. The authors are grateful to

the “Coordenacao de Aperfeigcoamento de Pessoal de Nivel Superior (CAPES)” for

financial support through project 88881.062172/2014-01 and the “Programa Ciéncia

sem Fronteiras.” RFCR is grateful to The Cuban Ministry of Agriculture and “Programa

de Salud Animal y Vegetal”, project P131LH003033 for facilities. We acknowledge the

assistance provided by Dr. P.M. Kirk and Dr. V. Robert through the Index Fungorum

and MycoBank websites. Dr. Lorelei Norvell’s editorial and Dr. Shaun Pennycook’s

nomenclatural reviews are greatly appreciated.

Literature cited

Castafieda-Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and South

America. 197-217, in: DW Li (ed.). Biology of microfungi. Springer International Publishing.

http://dx.doi.org/10.1007/978-3-319-29137-6_9

Flora do Brasil. 2020. <http://floradobrasil.jbrj.gov.br/>. Accessed on 25 April 2016.

Matsushima T. 1993. Matsushima mycological memoirs no. 7. Matsushima Fungus Collection,

Kobe, Japan.

Marvanova L, Om-Kalthoum Khattab S, Webster J. 1993. Mycofalcella calcarata, anam. gen. et sp.

nov. Nova Hedwigia 56: 401-408.

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

Biodiversity Series 9. 997 p.

MY COTAXON

July-September 2016— Volume 131, pp. 693-702

http://dx.doi.org/10.5248/131.693

Morphology and phylogeny of Cladosporium subuliforme,

causing yellow leaf spot of pepper in Cuba

BEATRIZ RAMOS-GARCIA’, TOMAS SHAGARODSKY’,

MARCELO SANDOVAL-DENIS’, YARELIS ORTIZ’, ELAINE MALOSSO?,

PHELIPE M.O. Costa}, JOSEP GUARRO‘4, DAvip W. MINTER‘,

DAYNET SoOSaA°, SIMON PEREZ-MARTINEZ’ & RAFAEL F. CASTANEDA-RUIZ?

"Instituto de Investigaciones Fundamentales en Agricultura Tropical ‘Alejandro de Humboldt’ (INIFAT),

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

? University of the Free State, Faculty of Natural & Agricultural Sciences, Dept. of Plant Sciences,

PO. Box 339, Bloemfontein 9300, South Africa

> Universidade Federal de Pernambuco, Depto. de Micologia/Laboratério de Hifomicetos de Folhedo,

Avenida da Engenharia, s/n, Cidade Universitaria, Recife, PE, 50.740-600, Brazil

‘Unitat de Micologia, Facultat de Medicina Ciéncies de la Salut, Universitat Rovira i Virgili,

43201 Reus, Spain

°CABI, Bakeham Lane, Egham, Surrey, TW20 9TY, United Kingdom

°Escuela Superior Politécnica del Litoral, ESPOL, CIBE,

Km 30.5, Via Perimetral, Guayaquil, Ecuador

“Universidad Estatal de Milagro (UNEMI), Facultat de Ingenieria, Milagro, Guayas, Ecuador

*CORRESPONDENCE TO: rfcastaneda@inifat.co.cu,rfcastanedaruiz@gmail.com

ABSTRACT—Cladosporium subuliforme is redescribed and illustrated from a specimen

collected from yellow leaf spots on pepper (Capsicum annuum) cultivated in high tunnels in

Cuba. Molecular phylogenetic analyses of morphologically close Cladosporium species are

provided.

Key worps—Capnodiales, hyphomycetes, plant disease, taxonomy, tropics

Introduction

Cladosporium Link (Bensch et al. 2010, 2012; Crous et al. 2007; Dugan

et al. 2004; Ellis 1971, 1976; Ho et al. 1999; Schubert et al. 2007) typified by

C. herbarum is a large cosmopolitan hyphomycete genus. IndexFungorum

694 ... Ramos-Garcia & al.

(2016) lists 777 names for Cladosporium, although several species have been

transferred to other genera (Bensch et al. 2010, 2012; Crous et al. 2007). Most

species are saprobes and can be found in almost all ecological niches and

substrates for easy spread through the air where they are common (Ho et al.

1999, Mullins 2001). Some species are pathogenic on cultivated plants (Ellis

1976; McKemy & Morgan-Jones 1990, 1992; Morgan-Jones & McKemy 1992;

Schubert & Braun 2004) and a few species are causal agents of human diseases

or disorders, mostly as opportunists (Hoog et al. 2011, Sandoval-Denis et al.

2015).

During an inspection of cultivated plants of Capsicum annuum (pepper) in

high tunnels at the INIFAT Campus, numerous yellow leaf-spotted plants were

found associated with a fungus identified as Cladosporium subuliforme. The

species is redescribed and illustrated below, and phylogenetic analysis of closely

related species is provided.

Material & methods

COLLECTION, ISOLATION, AND MORPHOLOGY— Samples of leafspots of pepper

were collected in high tunnels at the cultivated areas of the INIFAT Campus, Santiago

de Las Vegas, Cuba, and placed in paper bags, taken to the laboratory, and processed

according to Waller et al. (2002). The fungus was isolated from the substrate using a

single conidium isolation technique (Castafeda-Ruiz et al. 2000), grown on oatmeal

agar with adjusted pH 6.4, and incubated at 25 °C under alternating cycles of 12 h of

daylight and 12 h darkness. Mounts were prepared in PVL (polyvinyl alcohol) and

lactic acid and measurements were taken at x1000. Photomicrographs were obtained

with a Zeiss Axio Imager M1 light microscope (Carl Zeiss Meditec, Germany). The

pathogenicity test (Koch's postulates) of C. subuliforme on pepper plants was confirmed

after inoculation of healthy plants with C. subuliforme (INIFAT C13/100) following

the methodology proposed by Waller et al. (2002). Cultures obtained were deposited

in the INIFAT fungus collection, Santiago de Las Vegas, Cuba (INIFAT), in the

CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands (CBS), and in the

Faculty of Medicine, Rovira i Virgili University, Reus, Spain (FMR).

DNA EXTRACTION, SEQUENCING, AND PHYLOGENETIC ANALYSES — Total genomic

DNA was extracted from mycelia obtained from colonies growing on PDA using

FastPrep (MP Biomedicals, Santa Ana, CA) according to the manufacturer’s protocol.

DNA was quantified using the NanoDrop 3000 (Thermo Scientific, Madrid, Spain).

The primers ITS5 and ITS4 (White et al. 1990) were used to amplify the complete ITS

Fic 1. Cladosporium subuliforme. A, B. Infected pepper plant growing in a high tunnel;

C, D. Collected yellow leafspots on pepper;. E. INIFAT C13/100 colony on oatmeal agar after 10

days; F, G. Yellow leafspots on pepper after pathogenicity test inoculation.

Cladosporium subuliforme in Cuba... 695

F CE

696 ... Ramos-Garcia & al.

(ITS1+5.8S gene+ITS2) rDNA region; the primer pair LROR/LRS5 (Vilgalys & Hester

1990, Rehner & Samuels 1994) was used to amplify a fragment of the large subunit

(LSU) rDNA gene; the EF-728F/EF-986R primer pair was used for the translation

elongation factor la gene (EF-la) and the ACT-512F/ACT-783R primer pair for the

actin gene (Act) (Carbone & Kohn 1999).

Sequencing was performed in both directions using the same PCR primers at

Macrogen Europe (Macrogen, Inc., Amsterdam). Consensus sequences were obtained

using SeqMan version 7.0.0 (DNASTAR Lasergene, Madison, WI). An initial presumptive

generic identification of the isolates was performed based on BLAST searches of ITS and

LSU sequences in the GenBank (2016) and CBS (2016) databases. Multiple sequence

alignments of each locus were performed in MEGA version6 (Tamura et al. 2013) using

the ClustalW application (Thompson et al. 1994), refined with MUSCLE (Edgar 2004),

and manually adjusted where necessary. The alignments were complemented with a

total of 57 sequences from ex-type or reference strains obtained from GenBank and

selected on the basis of the sequence similarity results.

Phylogenetic reconstructions were made using maximum-likelihood (ML; MEGA

version 6) and Bayesian Inference (BI; MrBayes version 3.1.2) (Huelsenbeck &

Ronquist 2001). The best nucleotide substitution model (generalized time-reversible

model with gamma distribution and a portion of invariable sites [GTR+G+I] for the

three independent data sets) was estimated using MrModelTest version 2.3 (Nylander

2004) following the Akaike criterion. Phylogenetic analyses using ML were at first

made individually for ITS, EF-la, and Act and compared in order to assess for any

incongruent results between nodes with high statistical support. As no incongruences

were observed, the three loci were combined. For the ML analysis, nearest-neighbor

interchange (NNI) was used as the heuristic method for tree inference. Support

for the internal branches was assessed by a search of 1000 bootstrapped sets of data.

A bootstrap support value (BS) of 270 was considered significant. For BI analysis, two

simultaneous runs of 10,000,000 generations were performed, with samples stored

every 1000 generations. The 50% majority-rule consensus tree and posterior probability

values (PP) were calculated after discarding the first 25% of the samples. A PP value of

20.95 was considered significant.

Taxonomy

Cladosporium subuliforme Bensch, Crous & U. Braun, Stud. Mycol.

67: 77. (2010). Fics 1-3

SYMPTOMS ON HOST: subcircular, angular or irregular yellow leaf spot,

amphigenous, scattered, slightly necrotic at the centre. COLONIES on

oatmeal agar floccose, greenish gray to dark greenish olivaceous, margins

Fic 2. Cladosporium subuliforme (INIFAT C13/100). A. Ramoconidia and conidia;

B, C. Conidiophores, conidiogenous cells, ramoconidia, and conidia; D-F. Conidiogenous

cells, ramoconidia, and conidia. Scale bars = 10 um.

Cladosporium subuliforme in Cuba ... 697

698 ... Ramos-Garcia & al.

Fic 3. Cladosporium subuliforme (INIFAT C13/100).

Conidiophore, conidiogenous cells, ramoconidia, and conidia.

Cladosporium subuliforme in Cuba... 699

C. colocasiae CBS 386.64 cH 48067. HM4s310- H48555)_

Tio86 C. colocasiae CBS 119542 (IM148066- M1 48309.- HM 485

. tenuissimum CBS 125995 (M1 48197 -viMigsaa2, M1.

990.991 LC. tenuissimum CPC 13222 (Hiv 4g2t0-HMLa8455- HNILA8700

C. oxysporum CBS 125991. (HML48118- HML48362- HAT14360

c. oxysporum CBS 126351 (HM 148119- HM 148363. HMA 08)

“es C. cucumerinum CBS. 171. 52 (HM148072-4 FIML48316-

C. cucumerinum CBS 173. 54 (aM148074-8 HM 1483 18- side

C. subuliforme CBS 126500. (HIM 148196-HML48441-H

98/097

S097 C. angustisporum CBS 125983 (HML47995-1IM148236-

99/t C. cladosporioides. CBS. 113738 qisiiasooa- HIM148245- M4891)

C. cladosporioides CBS 112388 (HINT 49003-4 HIMT48244- HiM148490)

air C pseudocladosporioides CBS 125993 iiss finn 434024

9810.99 GC. funiculosum CBS 122129 ran as0%4- HIML48338 “M4898

x4 C. funiculosum CBS. 122128 M148093- HM 148337. HM148582) ees

C. herbarum CBS 121621 (r679363-2F679440-EF6795 16)

ee C. sphaerospermum CBS 193.54 (0Q780343-£U570261-EU570269)

C. halotolerans CBS 119416 (00780364-3N906939-£F 101397)

Cs. beticola CBS: 116456 «nrizi3is-Avs404o4-ays4o4ss)

0.05

Fic 4. Maximum likelihood tree inferred from combined ITS, EF-1a, and Act sequences showing

the phylogenetic relationships between the isolate INIFAT C13/100 and representatives of

Cladosporium spp. Numbers above the branches are bootstrap values (BS =70%) followed by

posterior probability values (PP 20.95). Fully supported branches (BS 100%/PP 1) are shown

in bold. Numbers between parentheses indicate the GenBank accession numbers for the ITS,

EF-la, and Act gene sequences. Sequences from type specimens marked in bold. ‘The scale bar

indicates the number of nucleotide substitutions per site. The tree was rooted with Cercospora

beticola CBS 116456.

pale olivaceous-brown or dull green, after 10 days or pulvinate, scattered,

amphigenous, yellow, 300-550 um diam. Mycelium superficial and immersed,

composed of septate, branched, verruculose, pale olivaceous brown hyphae,

2-4 um diam. CONIDIOPHORES macronematous, branched, erect, straight

or flexuous, cylindrical, unbranched or slightly branched toward the apex,

septate, olivaceous brown, smooth or verruculose near the base, 40-220 x

2-4 um. CONIDIOGENOUS CELLS polyblastic, terminal, sometimes intercalary,

integrated, cylindrical or oblong, somewhat subnavicular, 12-35 x 4 um with

several thickened and refractive conidiogenous loci, slightly denticulate.

Conidial secession schizolytic. RAMOCONIDIA primary and _ secondary,

cuneiform, subcylindrical, subnavicular, truncated at the base, 15-30 x 2-3.5

700 ... Ramos-Garcia & al.

um, smooth, olivaceous brown, dry. Conrpia blastocatenate, ellipsoidal,

ovoid, obovoid to subglobose, forming branched chains, unicellular, smooth,

pale olivaceous brown, dry, 3-7 x 3-5 um, truncated, slightly thickened and

refractive at the base or both ends.

SPECIMEN EXAMINED: CUBA. LA HaBana, Santiago de Las Vegas, INIFAT Campus,

in high tunnels, 22°58’N 82°20’W, on subcircular, angular, or irregular yellow leafspots

on Capsicum annuum L. (Solanaceae), 12-V1-2013, coll. T. Shagarodsky (Live cultures:

INIFAT C13/100 = CBS 140073 = FMR 14409; GenBank LN850753, LN850754,

LN850755, LN850756).

Note: The Cuban material has shorter ramoconidia and longer conidia than

those of the type material (ramoconidia 14-35 um long; conidia 2.5-12 um

long; Bensch et al. 2012), but otherwise fits well the original description.

Phylogenetic analyses Fig. 4

A preliminary identification of the isolate INIFAT C13/100 based on

sequence comparison of the ITS and LSU loci showed that it was related with

members of the Cladosporium cladosporioides complex. However, a species

identification was not possible using this approach given the poor resolution

of these loci.

A further phylogenetic analysis was performed based on the combined

ITS, EF-la, and Act sequences. The final alignment consisted of a total 973

characters (ITS 443; EF-la 325; Act 205), of which 378 were variable (ITS

89; EF-la 186; Act 103) and 182 were informative (ITS 13; EF-la 105; Act

64). The resulting phylogenetic tree (Fic. 4) showed that INIFAT C13/100

formed a strongly supported monophyletic group with the ex-type strain of

C. subuliforme (CBS 126500) (BS 100%, PP 1) with a sequence similarity of 100%

(ITS), 98.8% (EF-la), and 98.8% (Act). The clade representing C. subuliforme

clustered closely related with several species included in the C. cladosporioides

complex of Cladosporium: C. angustisporum, C. cladosporioides, C. colocasiae,

C. cucumerinum, C. oxysporum, and C. tenuissimum.

Therefore, the molecular analyses confirmed INIFAT C13/100 identification

as C. subuliforme. Our phylogeny agrees with previous work on Cladosporium

(Bensch et al. 2010, 2012); C. subuliforme clusters as a phylogenetically well-

defined species sister to C. angustisporum and C. cucumerinum. (Cladosporium

cucumerinum has also been reported from Capsicum annuum in South

America; Mendes et al. 1998). However, despite their genetic closeness,

the three species are easily differentiated based on the shape and length

of their conidiophores and conidia (Bensch et al. 2012). Two other species,

C. cladosporioides and C. oxysporum, also described from Capsicum species in

South America and Asia (Farr et al. 2016), are also related morphologically and

Cladosporium subuliforme in Cuba... 701

phylogenetically with C. subuliforme. Nevertheless, C. subuliforme differs from

C. cladosporioides by its long and narrow subuliform (subulate) conidiophores,

and its narrower conidiogenous loci and conidial hila, and can be distinguished

from C. oxysporum by its non-nodulose non-geniculate conidiophores (Bensch

et al. 2012).

Acknowledgments

The authors express their sincere gratitude to Dr. Xiu-Guo Zhang and Dr. De-Wei

Li for their critical review of the manuscript. The authors are grateful to Miss Mirta

Caraballo Fernandez and Sonia Alvarez for technical assistance. The authors are grateful

to the ‘Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)’ for

financial support through project 88881.062172/2014-01 and the ‘Programa Ciéncias

em Fronteiras. RFCR is grateful to the Cuban Ministry of Agriculture and “Programa

de Salud Animal y Vegetal’, project P131LH003033, for facilities. Dr. Lorelei L. Norvell’s

editorial review and Dr. Shaun Pennycook’s nomenclature review are greatly appreciated.

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http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

MY COTAXON

July-September 2016— Volume 131, pp. 703-715

http://dx.doi.org/10.5248/131.703

Boletus durhamensis sp. nov. from North Carolina

BEATRIZ ORTIZ-SANTANA |, ALAN E. BESSETTE * & OWEN L. MCCONNELL ?

'US Forest Service, Northern Research Station, Center for Forest Mycology Research,

One Gifford Pinchot Dr., Madison WI 53726

170 Live Oak Circle, Saint Marys, GA 31558

2808 Butner St., Durham, NC 27704

* CORRESPONDENCE TO: bortizsantana@fs.fed.us

ABSTRACT—A new bolete with cinnamon-brown pores, Boletus durhamensis, is described.

Collected in northern North Carolina, it is possibly mycorrhizal with Quercus spp.

Morphological and molecular characters support this taxon as a new species.

Key worps—Boletaceae, ectomycorrhizal fungi, oaks, taxonomy

Introduction

Studies on boletes in North Carolina started with Coker & Beers (1943).

Their work, which was concentrated in the Piedmont area of Orange County,

presented morphological descriptions of 68 Boletus, four Boletinus, and one

Strobilomyces species. Subsequent studies by Grand (1970a, b, c; 1981), which

provided more information about the occurrence, distribution, and host

associations of boletes in the mountains and Piedmont regions, added about

10 new records for the state. More recently as part of a workshop devoted to

boletes, Justice (2008) listed species from North Carolina based on Bessette et

al. (2000) and species reported by the Asheville Mushroom Club and North

American Mycological Association’s Wildacres Regional Forays.

In 2001, one of us (McConnell) found an unusual brown-capped bolete in

a hardwood forest at the Edison Johnson Recreation Center in Durham, NC.

The fruiting bodies consistently had brown pore surfaces in all developmental

stages, from the very youngest button through maturity. Boletes with brown

pore surfaces are rare in North Carolina, and despite careful evaluation of the

704 ... Ortiz-Santana, Bessette & McConnell

unknown bolete’s macrofeatures and a search of recent mycological literature, it

was not possible to identify the species. Ernst Both (Buffalo Museum of Science,

NY) recognized the material as an undescribed species, tentatively naming it

“Boletus durhamensis” based on its collection locality. Unfortunately, Both died

before he was able to publish these findings.

Binion et al. (2008) published one image of B. durhamensis with a description

(by McConnell), but the name was not validly published under the rules of the

International Code of Nomenclature. Since the first collection and description

of B. durhamensis were obtained when working in collaboration with Ernst

Both, and because new collections have been made, his provisional species

name is validated here.

Materials & methods

Morphological observation

Macroscopic descriptions are based on fresh and dried specimens, field notes, and

color photographs. Color terms are general approximations, while numerical color

designations are from Kornerup & Wanscher (1978). Macrochemical reactions were

determined using 10% NH,OH, 5% KOH, and 10% FeSO,. Microscopic structures were

observed with an Olympus BH-2 compound microscope; freehand sections of dried

fungal material were rehydrated in 70% ETOH and mounted separately in 3% KOH and

Melzer’s reagent. In the description of basidiospores, n = number measured, followed by

TABLE 1. Taxa included in the study, with their vouchers and GenBank accession

numbers. New sequences are indicated with bold font.

TAXON VOUCHER ITS LSU TEF

Boletus aereus REH-8721 — KF030339 KF030426

SU07 DQ131619 — —

B. amygdalinus 112605ba — JQ326996 JQ327024

src491 DQ974705 — —

B. calopus Bel — AF456833 JQ327019

UF1401 HM347645 — —

B. durhamensis BOS-885 KM675997 KM675998 —

Both-4561 KM675995 KM675996 KM668212

B. edulis BD380 EU231984 HQ161848 —

Bel — — JQ327018

B. luridus B12 — AF139686 —

AMB12640 KC734544 — —

B. pinophilus 42/93 — AF462359 —

isolate 2163 KC750240 — —

B. satanas Bs2 — AF042015 —

B. subvelutipes RV98-102 — AY612804 —

B. tenax REH-6871 — KF030320 KF030437

B. variipes

B. variipes var. fagicola

B. vermiculosus

Bothia castanella

B. fujianensis

Butyriboletus appendiculatus

B. regius

Caloboletus firmus

Sutorius australiensis

S. eximius

Tylopilus alboater

T. appalachiensis

T. atronicotianus

T. badiceps

T. felleus

T. ferrugineus

T. indecisus

T. microsporus

T. plumbeoviolaceus

“T. tabacinus”

Xerocomellus chrysenteron

X. cisalpinus

X. zelleri

Xerocomus fennicus

“X. perplexus”

X. pruinatus

X. ripariellus

X. subtomentosus

Chalciporus piperatus

C. pseudorubinellus

C. rubinellus

BD245

4249

BD190

222/97

MB03-053

MB03-067

HKAS-82694

HKAS-82693

Bap1

VDK-0429

11265

MG408a

MB06-060

Arora-13039

REH-9280

REH-9400

TH-8988

TDB-1206

TENN61182

Both s.n.

78206

AT2001011

HKAS-55832

JMP0093

MB06-053

98-98

HKAS-59661

MB06-056

HN-2295

HN-2295 (CFMR)

HKAS-56494

TENN60896

AT2005034

1B19980850

PDD94421

GS-10020

BB06304-Bv

REH-8724

KGP68

H126

MBO00-005

IB19961055

GR22465

strain Xs1

1549a-Q-6103

MB 04-001

2591

4302

2626

EU231958

EU231963

DQ867114

KM269196

HQ882194

KC584789

KM396278

KT339268

FJ596794

EU685114

EU819449

KX925216

FJ596906

KT271743

HM190074

DQ822794

JQ003657

KM248935

KM248949

KM248951

EU232003

JQ327014

DQ534646

DQ867117

KM269193

AF456837

KF030267

KF030278

JQ327005

JQ327004

AF139708

EU685110

KF030335

JQ326993

HQ326934

JQ326994

AF456820

KF112450

KF030350

AY612837

KX925217

KF112357

KF030354

AF514815

JQ924322

KF030271

AF514820

JQ003702

AF514825

AF514818

AF139716

DQ534648

KF030287

Boletus durhamensis sp. nov. (U.S.A.) ... 705

JQ327017

KF030421

JQ327025

KF030411

KF030408

JQ327031

JQ327029

KF030335

JQ327015

JQ327016

KF112225

KF030439

KF112172

KF030417

KF112171

KF030416

KF030438

JQ327035

GU187690

KF030441

706 ... Ortiz-Santana, Bessette & McConnell

the mean basidiospore lengths and widths + their standard deviations and the Q | value,

which represents the mean Q value + its standard deviation, where Q = length/width

ratio. Specimens examined are deposited in the CFMR herbarium. Herbarium acronym

follows Thiers (2016).

DNA extraction, PCR & sequencing

DNA sequences from two nuclear ribosomal DNA regions (LSU and ITS) and

one protein-coding gene (TEFl-alpha) of B. durhamensis were generated in the

present study. DNA extraction, amplification, and sequencing from dried specimens

of B. durhamensis were conducted at the Center for Forest Mycology Research in

Madison, following Palmer et al. (2008). The ITS region was amplified with primers

ITS1F (Gardes & Bruns 1993) and ITS4 (White et al. 1990), the 5’ end of the LSU

region was amplified using primer pair LROR/LRS5 (Vilgalys & Hester 1990) and

TEF1 was amplified using primer pair EF1-983/EF1-1567R (Rehner & Buckley

2005). For TEF PCR protocols see Minnis & Lindner (2013).

Phylogenetic analysis

DNA sequences were used primarily for molecular identification and were

compared with other sequences available in GenBank via BLASTn search (Benson et

al. 2013). DNA sequences were also used to infer the phylogenetic relationship among

B. durhamensis and other members of the family Boletaceae. Considering the BLASTn

search results and the main morphological characters of B. durhamensis, sequences of

species pertaining to the genera Boletus, Bothia, Butyriboletus, Caloboletus, Sutorius,

Tylopilus, Xerocomellus, and Xerocomus were retrieved from GenBank (TABLE 1).

Chalciporus piperatus (Bull.) Bataille, C. pseudorubinellus (A.H. Sm. & Thiers)

L.D. Gomez, and C. rubinellus (Peck) Singer were used as outgroup in the phylogenetic

analyses. Sequences were edited with Sequencher 4.8 (Gene Codes Corp., Ann Arbor,

Michigan), and aligned using MAFFT v.7 (Katoh & Standley 2013). The alignment

was manually adjusted using MacClade 4.08 (Maddison & Maddison 2002). ITS,

LSU, and TEF datasets were compiled and evaluated with Maximum likelihood (ML)

analysis run in RAxML server, v.7.7.1 (Stamatakis et al. 2008) under GTR GAMMA

model with 100 rapid bootstrap replicates.

Taxonomy

Boletus durhamensis B. Ortiz, Bessette & McConnell, sp. nov. PLATES 1-2

MycoBank MB 810115

Differs from Boletus vermiculosoides and B. vermiculosus by its lack of a bluing reaction

when its flesh is exposed, its partial veil covering the immature pores, and its smaller

basidiospores.

Type: USA. North Carolina: Durham Co., Edison Johnson Recreational Center, 11

August 2001, O.L. McConnell, Both 4561 (Holotype, CFMR, GenBank KM675995,

KM675996, KM668212).

EtryMoLoey: durhamensis refers to Durham, North Carolina, where this bolete was first

collected by mycologist Owen L. McConnell.

Boletus durhamensis sp. nov. (U.S.A.) ... 707

PiaTE 1. Boletus durhamensis (BOS 885). Basidiomata. Photo by OL McConnell.

Icongs: Macrofungi associated with oaks of eastern North America (Binion et al. 2008:

48).

Piteus 4-8 cm diam., hemispherical at first, becoming convex to broadly

convex at maturity; surface dry, subtomentose to subvelutinous, medium brown

to cinnamon-brown (near 5D7-5D5), becoming paler brown to yellowish

brown (near 4B6) in age; margin incurved when young, typically yellowish,

with a slight overlapping band of sterile tissue. FLEsH white, slightly tinged with

yellowish tan, unchanging when exposed; odor pleasant and somewhat fruity

when fresh, strong and disagreeable in dried specimens; taste not distinctive.

HYMENOPHORE tubular; tubes 2-6 mm deep, straw-colored (near 3B4), not

staining when bruised but becoming brown in dried specimens (6D7); pores

2-3 per mm in immature specimens, <1 mm diam. in mature specimens;

pore surface adnate to narrowly depressed near the stipe, uniformly deep

cinnamon-brown (near 6D6) in mature specimens, unchanging or slightly

darker brown when bruised; pores in immature specimens covered by a layer

of whitish hyphae (partial veil or conspicuously developed cheilocystidia), this

layer becoming greyish yellow (4B4-4C4) in dried specimens. STIPE 2.8-7.5

cm long, 1-2 cm broad, nearly equal or slightly enlarged downward, solid;

surface dry and smooth, with a very pale yellow ground color (3A3-4B3) that

typically becomes covered with cinnamon-brown (near 6D6) pruinosity, the

pruinosity being sparse on young boletes and heavier on mature ones, and the

stipe sometimes becoming vertically streaked with pruinosity; true reticulation

708 ... Ortiz-Santana, Bessette & McConnell

is often absent on young specimens, and when found on older stipes consisting

of a fine network typically restricted to the uppermost centimeter or two. Basal

mycelium white. STIPE FLESH solid, concolorous with the pileal flesh. SPORE

PRINT Olivaceous gray-brown (near 4D7) or olive-brown in fresh deposit.

BASIDIOSPORES 8.1-10.8 x 3.6-4.5 Um (n = 30, 9.33 + 0.96 x 3.9 + 0.35

um; Q = 2.40 + 0.15), fusoid to subcylindrical, smooth, grayish yellow in

KOH; pale yellowish or with brighter wall in Melzer’s. Bastp1a 22.5-27

(-41.4) x 7.2-8.1 um, mostly clavate, few cylindro-clavate (2) 4-sterigmate,

hyaline in KOH. Basrp1oLes 18-27(-38.7) x 6.3-8.1 um, mostly clavate.

PLEUROCYSTIDIA 24.3-48.6 x 5.4-11.7 um, most frequently lageniform, but

also fusoid to fusoid-ventricose or ventricose-rostrate, few cylindric, mostly

with rounded tip, smooth and thin-walled, hyaline or with yellow contents

in KOH, non-reactive in Melzer’s. CHEILOCYSTIDIA 22.5-52.2 x 5.4-9 um,

fusoid, ventricose-rostrate, smooth, thin-walled, hyaline or with pale yellow

contents in KOH, non-reactive in Melzer’s; terminal elements covering the

immature pores longer, 45-78.3 x 5.4-8.4 um, cylindrical to sub-constricted,

hyaline in KOH, non-reactive in Melzer’s. PILEIPELLIS a tangled layer of erect

to repent hyphae, 3.6-7.2 um diam., hyaline to grayish yellow in KOH; end

cells cylindrical. PILEUS TRAMA hyphae moderately to tightly interwoven,

4.5-19.8 um diam., hyaline in KOH, non-reactive in Melzer’s, smooth, thin-

walled. StiprTiPELLIs hyphae 3.6-11.7 um diam., parallel to subparallel to

interwoven, hyaline in KOH, yellowish in Melzer’s. CAULOCYSTIDIA 27-97.2 x

6.3-9 um, clavate, fusoid, cylindrical, sub-lageniform, in clusters (fasciculate),

hyaline in KOH, yellowish in Melzer’s, thin-walled. DERMATOBASIDIA present,

19.8-28.8 x 7.2-9 um, hyaline in KOH, contents non-dextrinoid to weakly

dextrinoid in Melzer’s. CLAMP CONNECTIONS absent.

MACROCHEMICAL REACTIONS: Pileipellis staining red-orange with KOH,

slightly dull vinaceous with NH,OH, and negative with FeSO,; flesh not

reacting (negative) with KOH, NH,OH, and FeSO.,.

ECOLOGY & DISTRIBUTION: solitary, scattered, or in groups on the ground in

a sparsely grassy area near a mixed broadleaf woods with willow oak (Quercus

phellos), white oak (Quercus alba), red maple (Acer rubrum), sweetgum

(Liquidambar styraciflua), and dogwood (Cornus florida); August; reported

only from the eastern Piedmont of North Carolina.

ADDITIONAL SPECIMENS EXAMINED: USA. NorTH CAROLINA: Granville Co., Town

of Butner, John Umstead Hospital picnic area, under white oak, 29 July 2014, O.L.

McConnell BOS 885 (CFMR; GenBank KM675997, KM675998); Durham Co., Edison

Johnson Recreational Center, under willow oak, 7 August 2014, O.L. McConnell BOS

886 (CFMR); Duke Forest Jogging Trail, October 1993, HN 2295 (DUKE, as “Tylopilus

tabacinus”).

Boletus durhamensis sp. nov. (U.S.A.) ... 709

PLATE 2. Boletus durhamensis (holotype Both 4561; BOS 855). A. basidiospores; B. basidia and

basidioles; C. pleurocystidia, some with yellow contents; D. cheilocystidia; E. cheilocystidia

covering the immature pores; F. caulocystidia and dermatobasidia; G. terminal elements of the

pileipellis. Scale bar = 10 um.

Additional characters of B. durhamensis based on dried specimens of collection

HN 2295 (initially misidentified as “Tylopilus tabacinus”): PILEUS <12 cm diam.,

plane, uplifted, smooth to finely velutinous, pale brownish orange (5C5). FLESH

cream. TUBES <11 mm deep, cinnamon-brown to raw sienna (6D7); pores

710 ... Ortiz-Santana, Bessette & McConnell

1-2 per mm yellowish brown (5D8), adnate to decurrent. Stipe 10 cm long,

3.5 cm diam., concolorous with pileus; finely reticulate at apex (11 mm from

apex), otherwise smooth; caespitose. BASIDIOSPORES 9-10.8 x 3.6-4.5 Um,

fusoid, smooth. Basip1a 20.7-26 x 8.1-9 um, clavate, 4-sterigmate, hyaline

in KOH. PLEuROCYSTIDIA 49.5-62.1 x 9-9.9 um, most frequently lageniform.

CHEILOCYSTIDIA 24.3-31.5 x 6.3-9.9 um, fusoid, ventricose-rostrate.

CoMMENTS— The cinnamon-brown or brown pores would place B. durhamensis

in Boletus sect. Luridi (Singer 1986), while the presence ofa hyphal layer covering

the immature pores (partial veil) and lack of any bluing reaction would place it

in B. sect. Boletus (Singer 1986). Within B. sect. Luridi, a brown pore surface is

also found in B. vermiculosoides A.H. Sm. & Thiers and B. vermiculosus Peck,

which differ by turning blue when their flesh is exposed, the absence of a partial

veil covering the immature pores, and their larger basidiospores. Within B. sect.

Boletus, the presence of the partial veil on immature pores is shared with several

species including B. aereus Bull., B. edulis Bull., B. pinophilus Pilat & Dermek,

and B. variipes Peck, all of which can be distinguished from B. durhamensis by

their possession of a true reticulum and larger basidiospores.

The presence of the partial veil covering the immature pores is also found

in Butyriboletus (Boletus sect. Appendiculati Lannoy & Estadés; Sutara 2014).

Sutara (2014), who refers to this veil as a layer of conspicuously developed

cheilocystidia, indicates its presence in several Butyriboletus species:

B. appendiculatus (Schaeff.) D. Arora & J.L. Frank, B. fechtneri (Velen.) Arora &

J.L. Frank, B. regius (Krombh.) Arora &J.L. Frank, B. roseogriseus (J. Sutara et al.)

Vizzini & Gelardi, and B. subappendiculatus (Dermek et al.) Arora & J.L. Frank.

Sutara also mentions that it is not clear whether this cheilocystidia arrangement

also occurs in American and Chinese Butyriboletus species, since neither Arora

& Frank (2014) nor Li et al. (2014) mention it in their descriptions. When

comparing Butyriboletus species with Boletus durhamensis, we found that most

of those species have pileus with reddish tones, a yellow hymenophore that

turns blue after bruising, and a stipe finely or strongly reticulate with reddish

tones, and somewhat larger basidiospores. Other species that also appear to

have this partial veil are Sutorius australiensis (Bougher & Thiers) Halling &

N.A. Fechner and S. eximius (Peck) Halling et al.; however both species differ

from B. durhamensis in their overall basidiocarp colors (which range from

reddish brown to purple brown and dark gray), ornamented stipes, reddish

brown spore print color, and larger basidiospores (Bessette et al. 2000, Halling

et al. 2012).

BLASTn searches on GenBank were performed independently with the

newly generated sequences. ITS and TEF BLASTn queries of B. durhamensis

Boletus durhamensis sp. nov. (U.S.A.) ... 711

Boletus_calopus_Bc1

Caloboletus_firmus_MB06_060

Boletus_amygdalinus_112605ba

100 ae Sutorius_eximius_REH9400

Sutorius_australiensis_ REH9280

100 Butyriboletus_appendiculatus_Bap1

Butyriboletus_regius_11265

92 Xerocomus_perplexus_ MBO0_ 005.

100 Boletus_tenax_REH6871

Xerocomus_subtomentosus_Xs1

Bothia_castanella_MB03_053

100 ; Xerocomellus_cisalpinus_PDD94421

Xerocomus_cisalpinus_AT2005034

Xerocomelius_zelleri_REH8724

85 Xerocomellus_chrysenteron_ HKAS 56494

75 Boletus_aereus_REH8721

70 100 Boletus_variipes_fagicola_4249

Boletus_edulis_Be1

100 Tylopilus_ferrugineus_MBO6_053

100

Tylopilus_badiceps_78206

Tylopilus_felleus_AT2001011

Tylopilus_microsporus_HKAS59661

Tylopitus_plumbeoviolaceus_MB06_056

100 Chalciporus_pseudorubinellus_4302

Chalciporus_piperatus_MB04_001

0.08

Figure 1. ML tree based on TEF sequences. Bootstrap values >70% are included above branches.

were uninformative at the generic level, with none of the searches exceeding

95% similarity with any available sequence. The nearest matches obtained were

isolates representing the genus Xerocomellus. When comparing LSU sequences,

the closest matches found were a sequence labelled as “Tylopilus tabacinus”

(isolate HN2295) and a sequence of Xerocomellus fennicus (Harmaja) Sutara

(isolate H126). In view of the search results and certain morphological

characters (e.g., spore print color, presence of partial veil on immature pores),

different bolete taxa were selected for additional sequence analyses to test the

relationship of B. durhamensis with other boletes.

The phylogenetic relationship was inferred using three different datasets

(ITS, LSU, TEF) and one phylogenetic analysis (ML). Results were based on

the topology of the best-scoring ML tree. Datasets comprised: (ITS) 26 ingroup

sequences, 578 characters; (LSU) 42 ingroup sequences, 875 characters;

712 ... Ortiz-Santana, Bessette & McConnell

84 Boletus_varlipes_fagicola_BD190

Boletus_variipes_BD245

80 Boletus_aereus_SU07

Boletus_edulis_BD380

Boletus_pinophilus_2163

Tylopilus_ felleus_JMP0093

ut Tylopilus_appalachiensis_TENN61182

Tylopilus_atronicotianus_Both_sn

100

Sutorius_eximius_TH8988

95 Boletus_appendiculatus_VKD_0429

Butyriboletus_regius_MG408a

Caloboletus_firmus_Arora13039

Boletus_amygdalinus_src491

400 Boletus_furidus_AMB12640

Caloboletus_calopus_UF 1401

100 Bothia_castanella_MB03_067

Bothia_fujianensis_ HKAS82693

100 Xerocomus_perplexus_MB00_005

Xerocomus_subtomentosus_1549aQ6103

100 Boletus_chrysenteron_TENN60896

89 Xerocomus_zelleri_KGP68

94 Xerocomus_cisalpinus_ BBO6_304_Bv_Pi_070507

Xerocomelius_cisalpinus_GS10020

100 Chaiciporus_rubinellus_2626

Chalciporus_piperatus_2591

0.05

FiGurRE 2. ML tree based on nrITS sequences. Bootstrap values >70% are included above branches.

(TEF) 24 ingroup sequences, 592 characters. The TEF analysis (Fic. 1) placed

B. durhamensis on an independent branch within a moderately supported clade

comprising species of Boletus s.s., Bothia, Butyriboletus, Caloboletus, Sutorius,

Xerocomellus, and Xerocomus; this clade appears as a sister clade of Tylopilus

s.s. The ITS and LSU analyses (Fics. 2, 3) clustered B. durhamensis with isolates

labelled as “Tylopilus tabacinus” (HN2295) in an independent clade and not

grouping with any of the other bolete genera mentioned above.

Because of the similarity of these LSU sequences, we obtained the specimen

of HN2295, which we examined morphologically and from which we generated

ITS and LSU sequences. Comparison of the morphological and sequence

data confirms that B. durhamensis and HN2295 are similar and that neither

Boletus durhamensis sp. nov. (U.S.A.) ... 713

- 100 [ Axon bs naticerss2620e

Tylopilus_ferrugineus_MB06_053

Tylopilus_indecisus_98 98

a ee en Aa Tylopilus_plumbeoviolaceus_MB06_056

Tylopilus_microsporus_HKAS59661

Tylopitus_alboater_TDB1206

Tylopilus_felleus_HKAS55832

Tylopilus_felleus_AT2001011

80 Boletus_aereus_REH8721

ia Boletus_pinophilus_42_93

Boletus_edulis_BD380

98 Boletus_variipes_BD245

Boletus_variipes_fagicola_4249

Xerocomus_cisalpinus_IB49980850

Xerocomus_cisalpinus_AT2005034

Xerocomellus_cisalpinus_PDD94421

Xerocomellus_chrysenteron_HKAS56494

Xerocomus_pruinatus_IB19961055

Xerocomellus_zelleri_REH8724

Xerocomus_fennicus_H126

Xerocomus_ripariellus_GR22465

100 — Boletus subvelutipes RV98_102

— Boletus_vermiculosus_222 97

Boletus_luridus_Bl2

Butyriboletus_regius_11265

Butyriboletus_appendiculatus_Bap1

Boletus_satanas_Bs2

Boletus_amygdalinus_112605ba

Xerocomus_perplexus_MB00_005

Boletus_tenax_REH6871

Xerocomus_subtomentosus_Xs1

Sutorius_eximius_REH9400

Sutorius_australiensis_REH9280

Bothia_fujianensis_HKAS_ 82694

Bothia_castanella_MB03_053

100

Boletus_calopus_Bc1

Caloboletus_firmus_MBO6_060

Chalciporus_pseudorubinellus_4302

Chalciporus_piperatus_MB04._001

0.05

FiGure 3. ML tree based on nrLSU sequences. Bootstrap values >70% are included above branches.

collection belongs to the genus Tylopilus or represents Tylopilus tabacinus

(Peck) Singer. Although similar to B. durhamensis in basidiocarp color and

overall appearance, T: tabacinus produces a pinkish brown to reddish brown

spore deposit, is often prominently reticulate over much of the stipe, has bitter

tasting flesh (Singer 1947), its pleurocystidia are dextrinoid in Melzer’s (Smith

& Thiers 1971), and it lacks cheilocystidia (Wolfe 1981).

Our phylogenetic analyses confirmed the results obtained through BLASTn

searches that B. durhamensis seems to be unrelated to any genus in the

Boletaceae currently represented in GenBank. Nonetheless, since several of

its morphological characteristics resemble species of Boletus s.l., we propose

its placement within Boletus. Recent molecular studies (Dentinger et al. 2010,

Nuhn et al. 2013, Wu et al. 2014) have demonstrated that several bolete genera

(e.g., Boletus, Tylopilus, and Xerocomus) are not monophyletic, and the creation

714 ... Ortiz-Santana, Bessette & McConnell

of new genera may differentiate the well-supported groups obtained within

each of them. Therefore, we suggest that more molecular data are still needed

to clarify the evolutionary relationship and history within Boletaceae. Not all

the described bolete species have been studied from a molecular perspective,

and there are characteristics, such as the partial veil on the immature pores or

the brown pore surface, that appear to have evolved independently in several

different lineages of the family. In conclusion, further molecular studies of

boletes are needed to determine whether B. durhamensis should remain in

Boletus or be transferred to a new and as yet undescribed genus.

Acknowledgements

The authors are very grateful to Drs. Roy E. Halling and Alfredo Justo for critical

review of the manuscript; we also thank Drs. D. Jean Lodge and Andrew M. Minnis for

valuable comments on an earlier version of the manuscript.

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Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koeltz Scientific Book, Koenigstein.

Smith AH, Thiers HD. 1971. The boletes of Michigan. University of Michigan Press, Ann Arbor,

USA.

Stamatakis A, Hoover P, Rougemont J. 2008. A rapid bootstrap algorithm for the RAxML web-

servers. Systematic Biology 75: 758-771. http://dx.doi.org/10.1080/10635150802429642

Sutara J. 2014. Anatomical structure in European species of genera Boletus s.str. and Butyriboletus

(Boletaceae). Czech Mycology: 66: 157-170.

Thiers B. 2016. [continually updated]. Index Herbariorum: A global directory of public herbaria and

associated staff. New York Botanical Garden's Virtual Herbarium. [Accessed: September/2014

http://sciweb.nybg.org/science2/IndexHerbariorum.asp ]

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

ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238-4246.

White TJ, Bruns T, Lee SS, Taylor J. 1990. Amplification and direct sequencing of

fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds).

PCR protocols: A guide to methods and applications. New York: Academic Press.

http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

Wolfe CB Jr. 1981. Type studies in Tylopilus. I. Taxa described by Charles H. Peck. Sydowia 34:

1992213;

Wu G, Feng B, Xu J, Zhu XT, Li YC, Zeng NK, Hosen MI, Yang ZL. 2014. Molecular phylogenetic

analyses redefine seven major clades and reveal 22 new generic clades in the fungal family

Boletaceae. Fungal Diversity 69: 93-115. http://dx.doi.org/10.1007/s13225-014-0283-8

MYCOTAXON

July-September 2016—Volume 131, pp. 717-721

http://dx.doi.org/10.5248/131.717

Diorygma fuscum sp. nov. from China

JIAN LI & ZE-FENG JIA*

College of Life Sciences, Liaocheng University, Liaocheng, 252059, China

* CORRESPONDENCE TO: Zffia2008@163.com

ABSTRACT—A new lichen species, Diorygma fuscum, is described from Fujian Province,

China. It is characterized by the conspicuous open immersed lirellae with densely white

pruinose discs, the hyaline to brownish muriform ascospores sized 40-60 x 12-18 um, and

the presence of stictic acid. The type specimen is deposited in HMAS-L.

Key worps—Ascomycota, Ostropales, Graphidaceae, taxonomy

Introduction

Diorygma Eschw. (Graphidaceae) is mainly a tropical to subtropical genus

(Staiger 2002; Kalb et al. 2004), characterized by its inconspicuous pseudocortex,

lirellate ascocarps with a pruinose disc, branched or anastomosing paraphyses

with a thick gelatinous wall, Graphis-type 1-8-spored asci, ascospores that are

transversely septate with lenticular spore locules or muriform, and the presence

of norstictic, stictic, and/or protocetraric acid. At present, more than 50 species

are known worldwide (Kalb et al. 2004; Caceres 2007; Archer 2009; Makhija

et al. 2009; Sharma & Makhija 2009a, b; Sharma & Khadilkar 2012; Lima et al.

2013; Feuerstein et al. 2014; Sutjaritturakan et al. 2014).

Nine Diorygma species have been reported from China: D. hieroglyphicum,

D. hololeucum, D. junghuhnii |= Graphina mendax], D. macgregorii,

D. megasporum, D. pachygraphum, D. poitaei [= Graphina virginea],

D. pruinosum, and D. soozanum [=Graphina soozana] (Jia & Wei 2016, Kalb

et al. 2004, Lamb 1963, Meng & Wei 2008, Nakanishi et al. 2003, Thrower

1988, Wang Yang & Lai 1973, Wei 1991, Wei et al. 2013, Wu & Qian 1989,

Zahlbruckner 1933). Here we describe a new species collected in Fujian

Province, China, for which we propose the name Diorygma fuscum.

718 ... Li & Jia

Materials & methods

Specimens are housed in Herbarium Mycologicum Academiae Sinicae-Lichenes,

Beijing, China (HMAS-L). A dissecting microscope (Olympus SZX12) and a light

microscope (Olympus BX51 & Nikon Eclipse-55i) were used for the morphological and

anatomical studies. Measurements and illustrations were taken from the manual cross-

sections of fruit bodies in 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 (Culberson & Kristinsson 1970; Culberson 1972; White & James 1985).

Taxonomy

Diorygma fuscum Jian Li bis & Z.F. Jia, sp. nov. Pid

FUNGAL NAME FN 570263

Differs from Diorygma pruinosum by its smaller ascospores and the presence of stictic

acid.

Type: China. Fujian Province, Jianou City, Fangdao Town, Wanmulin, 27°02’N 118°08’E,

alt. 310 m, on bark, 3/VI/2007, Q.F. Meng FJ1280 (Holotype, HMAS-L 137193).

ErymMo.ocy: The epithet refers to the brownish mature muriform ascospores.

THALLUS crustose, pale grey to olive-grey, 60-100 um thick, surface uneven to

slightly rugose or warty, without soralia or isidia; PsEUDOCORTEX indistinctly

developed, 5-10 um thick, partly lacking; ALGAL LAYER 30-40 um thick;

MEDULLA poorly developed.

ASCOCARPS numerous, lirellate, often flexuous branched, immersed in the

thallus when young, becoming open and + raised when older, whitish, powdery,

acute or rounded at the ends, 1-4 x 0.3-2 mm; pDisc surrounded by entire

(in young apothecia) raised thalline margins, opened, rarely + convex, with

thick and white pruina, often showing fissures when older; EXCIPLE divergent,

laterally uncarbonized, rudimentarily developed, consisting of a weakly and

irregularly or brownish hyphal tissue intermingled with parts of the substrate,

carbonization sometimes restricted to the basal position; HYMENIUM 100-180

uum high, not inspersed, I+ weakly bluish violet; EPrrHECIUM usually distinctly

developed, consisting of intermingled anastomosing, hyaline or brownish

paraphysis tips with short + globular cells, hyaline granules, and dead hyphae;

PARAPHYSES 1-2 um diam., with a thick gelatinous wall, often anastomosing,

especially in the upper part of the hymenium and near the asci. ASCOSPORES

8 per ascus, hyaline to brownish (when mature), muriform, spore locules of

equal size, often arranged in rows, 10-14/3-4-locular, 40-60 x 12-18 um,

I+ violet, with thin halo.

PYCNIDIA not seen.

CORTICOLOUS.

Diorygma fuscum sp. nov. (China) ... 719

PiaTE 1. (holotype, HMAS-L 137193). A. Thallus with apothecia; B. Cross-section of

apothecium; C. Asci with ascospores, showing the thickened top of ascus; D. Asci with hyaline

to brownish ascospores. E. Ascus with brownish ascospores. F. Mature ascospore with thin halo.

Scale bars: A = 1 mm; B = 100 um; C-F = 20 um.

CHEMISTRY: Stictic acid (major), constictic, hypostictic and hypoconstictic

acids (minor, trace or absent).

ADDITIONAL SPECIMENS EXAMINED: CHINA. FUJIAN PROVINCE, Jianou City, Fangdao

Town, Wangmulin, alt. 400 m, 1/V1I/2007, Q.E Meng FJ1059 (HMAS-L 137198); alt. 420

m, 1/V1/2007, Q.F. Meng FJ860 (HMAS-L 137197); alt. 310 m, 3/VI/2007, Q.F Meng

FJ697 (HMAS-L 137196), FJ1278 (HMAS-L 137195), FJ1279 (HMAS-L 137194); alt.

300 m, 3/VI/2007, J. Li FJ630 (HMAS-L 137200); alt. 540 m, 2/VI/2007, J. Li FJ1066

(HMAS-L 137199).

Eco.oey. Diorygma fuscum grows on bark in relatively dry and open situations

from lowland primeval forests in subtropical area of China. Associated lichens

include Graphis hossei Vain. and other species of Graphidaceae and Lecanora.

REMARKS: Diorygma fuscum is morphologically similar to D. pruinosum

(Eschw.) Kalb et al., which differs in having 1-spored asci, larger ascospores

(95-170 x 19-50 um), and the presence of protocetraric acids. The new lichen

720 ... Li & Jia

species also resembles D. erythrellum (Mont. & Bosch) Kalb et al. and D. poitaei

(Fée) Kalb et al. in producing 8-spores per ascus and similarly sized muriform

ascospores (30-65 x 12-20 um in D. erythrellum; 40-65 x 10-18 um in

D. poitaei), but differs in having opened discs, a slightly carbonized proper

exciple at the base, and the presence of stictic acid (major); D. erythrellum

differs in containing norstictic acid (major), connorstictic acid (minor or

trace), and stictic acid (trace or absent), while D. poitaei contains hypostictic

and hypoconstictic acids (major) and o-acetylhypoconstictic, constictic, and

stictic acids (minor, trace or absent) (Kalb et al. 2004).

Acknowledgements

This study was supported by the National Natural Science Foundation of China

(31270066, 31093440, 31493010 & 31493011). The authors are grateful to Dr. Santosh

Joshi (CSIR-National Botanical Research Institute, Lucknow, India) and Dr. Hua-Jie

Liu (College of Life Sciences, Hebei University, China) for reading and improving the

manuscript, and for acting as presubmission reviewers.

Literature cited

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Caceres MES. 2007. Corticolous crustose and microfoliose lichens of northeastern Brazil. Libri

Botanici 22. 168 p.

Culberson CF. 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, Kristinsson H. 1970. A standardized method for the identification of lichen products.

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Feuerstein SC, Cunha-Dias IPR, Aptroot A, Eliasaro S, Caceres MES. 2014. Three new Diorygma

(Graphidaceae) species from Brazil, with a revised world key. The Lichenologist 46(6): 753-761.

http://dx.doi.org/10.1017/S002428291400036X

Jia ZE, Wei JC. 2016. Flora lichenum sinicorum - Vol.13, Ostropales (1). Science Press, Beijing. 210 p.

Kalb K, Staiger B, Elix JA. 2004. A monograph of the lichen genus Diorygma - a first attempt.

Symbolae Botanicae Upsalienses 34 (1): 133-181.

Lamb IM. 1963. Index Nominum Lichenum inter Annos 1932 et 1960 Divulgatorum. The Ronald

Press Company, New York. 809 p.

Lima EL, Maia LC, Aptroot A, Caceres MES. 2013. New lichen species from Vale do Catimbau,

Pernambuco, Brazil. The Bryologist 116(4): 327-329.

http://dx.doi.org/10.1639/0007-2745-116.4.327

Makhija U, Chitale G, Sharma B 2009. New species and new records of Diorygma (Graphidaceae)

from India: species with convergent exciples. Mycotaxon 109: 379-392.

http://dx.doi.org/10.5248/109.379

Meng QF, Wei JC. 2008. A lichen genus Diorygma (Graphidaceae, Ascomycota) in China.

Mycosystema 27: 525-531. http://dx.doi.org/10.1017/S002428291300025X

Nakanishi M, Kashiwadani H & Moon KH. 2003. Taxonomical notes on Japanese Graphidaceae

(Ascomycotina), including some new combinations. Bulletin of the National Science Museum

(Tokyo), Series B (Botany) 29(2): 83-90.

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Sharma B, Khadilkar P. 2012. Four new species of Diorygma from India. Mycotaxon 119: 1-10.

http://dx.doi.org/10.5248/119.1

Sharma B, Makhija U. 2009a. Four new species in the lichen genus Diorygma. Mycotaxon 107:

87-94. http://dx.doi.org/10.5248/107.87

Sharma B, Makhija U. 2009b. New species and new reports of Diorygma (lichenized Ascomycotina,

Graphidaceae) from India. Mycotaxon 109: 209-217. http://dx.doi.org/10.5248/109.209

Staiger B. 2002. Die Flechtenfamilie Graphidaceae: Studien in Richtung einer natiirlicheren

Gliederung. Bibliotheca Lichenologica 85: 1-526.

Sutjaritturakan J, Saipunkaew W, Boonpragob K, Kalb K. 2014. New species of Graphidaceae

(Ostropales, Lecanoromycetes) from southern Thailand. Phytotaxa 189(1): 312-324.

http://dx.doi.org/10.11646/phytotaxa.189.1.22

Thrower SL. 1988. Hong Kong lichens. Urban Council Publication, Hong Kong. 193 p.

Wang Yang JR, Lai MJ. 1973. A checklist of the lichens of Taiwan. Taiwania 18(1): 83-104.

http://dx.doi.org/10.6165/tai.1973.18.83

Wei JC. 1991. An enumeration of lichens in China. International Academic Publishers, Beijing.

278 p.

Wei JC, Jia ZF, Wu XL. 2013. An investigation of lichen diversity from Hainan Island of China

and prospect of the R & D of their resources. Journal of Fungal Research 11(4): 224-238

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White FJ, James PW. 1985. A new guide to microchemical techniques for the identification of

lichen substances. British Lichen Society Bulletin 57 (Suppl.): 1-41.

Wu JN, Qian ZG. 1989. Lichens. 158-226, in Xu BS (ed.): Cryptogamic flora of the Yangtze Delta

and adjacent regions. Shanghai Scientific & Technical Publisher, Shanghai (in Chinese).

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Vegetabilis 31: 194-224. http://dx.doi.org/10.1002/fedr.19330311108

MYCOTAXON

July-September 2016—Volume 131, pp. 723-731

http://dx.doi.org/10.5248/131.723

BOOK REVIEWS AND NOTICES’

LORELEI L. NORVELL

Pacific Northwest Mycology Service, Portland OR 97229-1309 USA

ABSTRACT—Books reviewed include: GENERAL—Mushrooms of the Redwood Coast

(Siegel & Schwarz 2016); BasiplomycEeTES—The genus Mycena s.]. (Arne Aronsen &

Thomas Laess@e).

NOAH SIEGEL and CHRISTIAN SCHWARZ

GENERAL

Mushrooms of the Redwood Coast—a

comprehensive guide to the fungi of coastal MUSHROOMS

northern California. By Noah Siegel & of the Redwood Coast

Christian Schwarz. 2016. Ten Speed Press

[Crown Publishing Group], Berkeley CA <www.

A Comprehensive Guide

to the Fungi ef Coastal Northern California

randomhouse.com/crown/tenspeed/>. 608 p. ~855

color photos, ISBN 978-1-60774-817-5 US$35 (soft

cover), ISBN 978-1-60774-817-2 $18.99 (e-book)

Two and a half decades of sequence analyses have ‘steam-rollered’ rapid name

changes in fungi, often leaving mushroomers and field taxonomists perplexed

and decidedly behind the nomenclatural curve. There was a time in the not

too distant past that comprehensive regional field guides limited coverage to

relatively few species representing the larger and more commonly encountered

macrofungi. Fortunately, the recent upsurge of beautifully illustrated and

nomenclaturally up-to-date regional field guides (e.g., Trudell & Ammirati

2009, Desjardin & al. 2015) have made life for west coast North American

* Book reviews or books for consideration for coverage in this column should be sent to the

Editor-in-Chief <editor@mycotaxon.com> 6720 NW Skyline, Portland OR 97229 USA.

724 ... MYCOTAXON 131(3)

mushroomers a lot easier by including revised nomenclature along with

striking photos of less commonly encountered fungi.

MUSHROOMS OF THE REDWOOD CoasT [advertised by the press as

“A comprehensive and user-friendly field guide for identifying the many

mushrooms of the northern California coast, from Monterey County to the

Oregon border” is a glorious addition to this new pantheon of field guides. The

print copy is a hefty 24 x 19 x 3.8 cm and provides full treatments of over 750

species and references hundreds more. Unfortunately the attractive paper cover

on my copy soon separated from the glue strip on the front page, foreshadowing

a duct-tape repair in the not too distant future. Glossy inside pages display the

sharp clear photos to their best advantage.

The 22-page introductory text is necessarily brief but more than adequately

covers the basics. A one-page introduction entices the non-initiated to further

coastal forest explorations and precedes an outline of mushroom morphology,

fungal ecology, a life cycle diagram, and human-fungal relationships. Six

pages devoted to the ‘redwood coast’ offer a map of the five regions (Far North

Coast; North Coast; San Francisco Bay Area & East Bay Hills; Santa Cruz &

Peninsula Mountains; and Monterey County and southernmost redwoods) and

tree descriptions and superb photos of the ectomycorrhizal northern conifers

[Sitka spruce, western hemlock, grand fir], ectomycorrhizal hardwoods [3

live oaks, two deciduous oaks, tanoak, chinquapin, madrone, manzanita],

widespread ectomycorrhizal conifers [Douglas-fir, five pine species], and

non-ectomycorrhizal trees [the coast redwood itself (!), Monterey cypress,

California bay laurel). Two and a half pages are dedicated to collection

practices: finding, collecting, and identifying mushrooms, making spore prints,

a list of necessities (basket, field notebooks, tackle boxes, waxed paper bags

[for which I would substitute aluminum foil], hand lenses, KOH, camera, and

food dehydrator), making collections, photographic tips, and collecting for the

table. The introduction ends with an excellent overview on “How to read the

species descriptions” (the sort of instruction too often ignored by eager would-

be identifiers), “General format of the species descriptions” (see below), an

explanation of “How to use the pictorial key to the major sections” followed by

the six-page pictorial key itself.

Photos paired with descriptions serve as the sole key to species, making

this volume definitely geared to the browse-and-point mode of identification.

While I do miss more formal keys to species, participation in enough forays

and mushroom collecting expeditions suggests that a photo matching routine

is perhaps the wisest and most efficient route for would-be identifiers lacking

microscopes and reagents—particularly when so many run into dead ends using

Book Reviews ... 725

written keys. The pictorial key is based on visual appearance, primarily stature

and spore print color. Given the rapid name changes and generic shifts, this

works relatively well, although my ‘generically’ trained brain sometimes found

itself confused at discovering a species in one section I anticipated in another.

The authors sub-divide some of the more unwieldy sections: for example, the

notorious 97-page ‘white-spored multitude’ (divided pictorially into ‘large’

vs. ‘small’) is synoptically apportioned into groups A-N, while the well-

populated bolete section is helpfully sorted according to its 16 long-accepted

(Boletus, Leccinum, Suillus) and quite recent (Butyriboletus? Rubroboletus?)

genera. The other 27 sections include chanterelles & gomphoids; Amanita;

Cystoderma + Cystodermella; Lepiota & allies; Agaricus + Melanophyllum;

dark-spored mushrooms; brown-spored decomposers; mycorrhizal brown

spored mushrooms (3 sections: Inocybe, Hebeloma, Phaeocollybia); Cortinarius;

Entoloma (2 sections: large, small); Pluteus and allies; Russula; Lactarius; waxy

caps (2 sections: I, II); pleurotoids; gilled boletes; polypores and allies; shelflike,

conklike, and rosette-forming polypores; crusts; toothed; corals; clubs;

puffballs, earthballs & earthstars; stinkhorns; bird’s nests; truffles; jellies; morels,

false morels & elfin saddles; and cup fungi. I was pleased to see a gratifyingly

large number of small to tiny mushrooms over which I have puzzled for years.

The species represented seem comprehensive, although one local forayer has

observed that as the authors collected during Californias recent and very

long drought, there are undoubtedly a great many species they never had the

opportunity to capture. Nonetheless I was personally charmed by the inclusion

of six beautifully photographed Phaeocollybia species (all accurately identified)

with reference to 14 others!

Each section outlines its genera with their diagnostic characters. Individual

species treatments each provide a color photo, Latin name and authority, often

a “common” name (some actually common, the remainder useful inventions),

and a formal technical description of cap, gills/other fertile surfaces, stipe, veil

(including partial veil & volva), flesh, odor & taste, KOH or other chemical

reactivity, spore deposit, and microscopic details (always spore shape & size;

other diagnostic characters when helpful). Noting that the microcharacters

provided are minimal, Siegel & Schwarz wryly observe: “There's still much to

be learned about the range and significance of variation in microscopic features

of mushrooms—our data are representative but not definitive. Please seek out

measurements from other sources and tell us what you learn!” Each treatment

is rounded out with information on ecology (more comprehensive than usually

found in field guides), edibility, the all-important COMMENTs paragraph, and

synonyms, misapplications, and other nomenclatural notes.

726 ... MYCOTAXON 131(3)

Each major species entry (1-2 per page) consists of a photo paired with

text. The photography is stellar throughout, expecially noticeable in the

introductory three large full-page plates and numerous half-page photos; the

smaller (~1/6 page) photos accompanying most species descriptions appear

equally clear, but fans (Iam one) of the volume should consider also purchasing

the online version so as to zoom these smaller photos for details not easily seen

on the printed page. With no photographers credited (either by the photos or in

the acknowledgments), we should assume that the two author-photographers

deserve kudos for capturing so many fine photos in the field.

The authors have adopted Arora’ (1986) term ‘group’ (e.g., Lyophyllum

decastes group, Xerocomus subtomentosus group, Scutellinia scutellata group)

and have introduced placeholder names (e.g., Craterellus tubaeformis, Hygrocybe

punicea, Russula cyanoxantha, here all followed by ‘sensu CA’), particularly useful

when depicting taxa known to differ from their European sobriquets but which

are yet to be described. [What made Arora’s ‘group’ so wonderful for so many

years is that it satisfied those anxious to name a specimen while simultaneously

indicating that more research is needed to suss out the ‘real’ identification. ]

No doubt equally satisfying to the ‘namers’ among us is that the authors are

not shy about providing provisional names by enclosing unpublished epithets

(e.g., Dendrocollybia “pycnoramella”, Leptonia “Ruby Grapefruit”, Xerocomellus

“diffractus”) and new combinations (“Phaeoclavulina” myceliosa, “Gliophorus”

flavifolius, “Xerocomellus” mendocinensis) in quotation marks. Exceedingly

helpful to the field mycologist, this also suggests a degree of nomenclatural

trust on the part of Siegel & Schwarz that unethical competitors will not rush to

publish the names ahead of them.

Regarding misapplications and synonomies, the authors note, “Californian

mushrooms are often quite different in morphology, microscopic features,

ecology, and genetics from their European namesakes, but until a new name is

published, the most effective way of referring to them may be to use an epithet

we know to be incorrect. We recommend taking a practical view of things:

incorrect names will be used to convey information, and if they succeed in

doing so without creating any new confusion, don't worry too much about it.”

Thus noting that Cortinarius percomis is a European name long misapplied

for C. citrinifolius, Neoalbatrellus (Albatrellus) caeruleoporus is an eastern

North American name misapplied for N. subcaeruleoporus, and Lactarius

vinaceorufescens & L. chrysorrheus are both eastern North American names

misapplied for L. xanthogalactus help link a familiar image and the new correct

name together. Synonyms serve a similar function (e.g., Atheniella adonis =

Mycena adonis; Lichenomphalia umbellifera = Omphalia ericetorum [somewhat

Book Reviews ... 727

baffling to me, given that the US government's Northwest Forest Plan listed

OMPHALINA ericetorum]; Mycetinus copelandii = Marasmius copelandii, with

a highly informative note citing recent genetic work that supports the species

in Gymnopus; Bolbitius titubans = B. vitellinus). Some ‘newer’ names lack

synonyms—the earlier placement of Tapinella atrotomentosa in Paxillus and

Annulohypoxylon thouarsianum in Sphaeria are not noted—but such omissions

are few. In general, I greatly appreciated finding so many ‘hot-off-the-press’

names with origins in multi-gene sequence analyses; these no doubt owe their

presence in the volume to the careful review of Else Vellinga, whom Siegel &

Schwarz thank first in their acknowledgments.

The volume closes with a list of future directions (mycofloristics,

conservations, climate change, biogeography, systematics, sequence database

quality, evolutionary and ecological synthesis), a 3-page glossary, resources for

the mycophile, bibliography, general index, and genus & species index.

Overall the field guide is engagingly written and with understated humor

highly appreciated here). I highly recommend this book.

Arora D. 1986. Mushrooms demystified (2"¢ edn.). Ten Speed Press, Berkeley

CA. 959 p.

Trudell S, Ammirati J. 2009. Mushrooms of the Pacific Northwest. Timber

Press, Portland OR. 351 p.

Desjardin DE, Wood MG, Stevens FA. 2015. Timber Press, Portland OR. 560 p.

BASIDIOMYCETES

The genus

Fungi of Northern Europe—Vol. 5. The genus

Mycena s.l. By Arne Aronsen & Thomas Lessge.

2016. Svampetryk <www.svampe.dk>, Narayana Press,

Gylling, Denmark. 373 p. ~378 color photos, ~116

micro-drawings. ISBN 978-87-983581-2-1. Price 3

Society members). .

by Arne Aronsen & Thomas Lassae

How monographs have changed! Eighteen years ago I regularly labored far into

the night over scores of Mycena collections while poring over microdrawings

in my venerable Alexander Smith (1947), Maas Geesteranus (1992), Rexer (1994),

728 ... MYCOTAXON 131(3)

and unpublished keys and notes shared by Scott Redhead and riffling through

the far-too-few color plates in Breitenbach & Kranzlin (1991). Not until Robich

(2003) landed on my library shelves did I have a full-color monograph dedicated

to these beautiful agarics—and Mycena is a genus that wears its colors very well,

indeed.

Aronsen & Lzessge’s new volume is as beautiful as the mushrooms it depicts.

Its size, large enough to be encyclopedic yet small enough to take to (if not into)

the field (23 x 17 x 3 cm), and its durable hard cover surrounds semi-glossy

easy-to-read glare-free pages bearing text and glossy photos that are especially

sharp and clear. Photo credits, most often placed unobtrusively to the right of

each photo, cite not only photographer and country but also collection number,

a wonderful addition I’ve not noticed in other monographs. Of the 160 mycenas

recorded from Europe, 115 are fully described here.

Authors who cover a genus in the broad sense must contend with when (or

whether) to fit the nomenclature to taxonomic breakthroughs. As Aronsen &

Leessoe note in their preface (written in May 2016), “It has become increasingly

clear that Mycena is not a monophyletic entity, and we have thus chosen to

title this volume Mycena sensu lato.... Most taxa have been accepted or left

within the genus Mycena but the two satellite genera, Roridomyces [Rexer 1994]

and Resinomycena [Redhead & Singer 1981] have become more or less well

established and are also ad interim recognized here, whilst newer introductions

such as Atheniella (for M. adonis a.o.) [Redhead 2012] and Phloeomana (for M.

speirea a.o.) [Redhead 2013] have not been formally recognized in this book,

although we accept that the genus Mycena as here circumscribed will have to be

split. The old generic name Prunulus, reintroduced by Redhead & co-workers

for Mycena sect. Calodontes, is likewise not treated as an independent genus in

this treatment.” In their historical recapitulation, the authors also draw attention

to the on-going battle between lumpers and splitters: [“... unlike Kihner (1923),

Singer (1938)] excluded some of the inamyloid species, and transferred them to

his newly erected genus Hemimycena.... He, likewise, transferred another group

of species (mainly with huge pleuro- and cheilocystidia and + ornamented

spores to the genus Mycenella. He was in other words a splitter, while Kithner

tended to be a lumper. In the era we live in now, the splitters are having the

upper hand..... Within the temperate world of the Northern Hemisphere the

baton was taken over by Rudolph Arnold Maas Geesteranus.... Although he

rarely worked with living specimens, he, nevertheless, produced a monumental

and long lasting monograph. He excluded Mycena rorida from the genus

mainly based on the structure of the pileipellis, and Rexer (1994) erected

the genus Roridomyces to accommodate this and six other species. Another

Book Reviews ... 729

deviating Mycena, known under a host of names ... was also excluded by Maas

Geesteranus and accepted as Resinomycena saccharifera....”

In their later discussion on phylogeny, the authors offer further generic

insights: “...it has become clear that Mycena as understood by Kihner, Singer,

and Maas Geesteranus ... is polyphyletic. Unfortunately, various efforts have

not so far [led] to a paper that provides a useful overview of the phylogenetic

position of the taxa that are and once were accepted within the genus. Early

poorly resolved trees have been used to create two new genera, and one old genus

has been accepted by some. ... Atheniella for e.g. M. adonis and Phloeomana

for M. speirea were created by Redhead and collaborators (Redhead 2012) ina

very brief contribution. Redhead (2016a, b, c), equally brief, added four more

species to Phloeomana ... and in Gminder (2016) another Atheniella was added.

It will be interesting to see whether a situation that parallels that seen in Boletus

s..—hyper splitting—will be the trend in mycenoid fungi. In this context

we have had to be conservative since no good phylogenetic trees are publicly

available.” In part, these comments address a controversy revolving around the

recent trend toward exceedingly brief publications that fulfill the letter of the

International Code of Nomenclature, if not the spirit, by presenting a name,

diagnosis, holotype, and relatively little discussion placing the new taxon in

context. While I remain at heart a splitter (finding smaller monophyletic genera

easier to master than larger polyphyletic ones), this move toward minimalist

publishing is hardly helpful. Unfortunately additional sequence analyses or

other as-yet untapped scientific innovations are unlikely to settle the splitter

vs. lumper controversy. When it comes down to it, taxonomy seems always to

remain a matter of opinion.

The formal introduction and a brief paragraph on (very) basic materials and

methods (noting that the names used more or less agree with those accepted

by Index Fungorum and MycoBank) are followed by 23 pages devoted to “The

genus Mycena s.l? ‘This chapter lists accepted generic synonyms followed by

a generic description. Macroscopic features are accompanied by stunning

detailed photos of ephemeral characters (including one of the best photos

of a separable elastic gill thread I’ve seen) and the discussion of important

microscopic features includes sketches of cheilocystidial types (p. 19) and

pileipellis types and setae. [Drawings throughout the book are elegantly

minimalist but get the job done.] “Ecology and habitat” pairs six pages of

excellent plates of representative species with habitats (e.g., M. chlorantha in

dunes, M. aetites on lawns, M. clavicularis in pine forests). Brief paragraphs on

conservation, seasonality, and development (one sentence referring to Rexer’s

thesis), cytology & culture studies, bioluminescence, toxicity, and parasites,

and phylogeny (see above) follow.

730 ... MycoTAxon 131(3)

Next comes a brief discussion of systematic arrangement & species concepts

and a list of the 32 sections (with species covered), both according to Maas G.

Particularly helpful are the six pages of photos and brief descriptions of

genera often confused with Mycena. Discussed here are two covered in the

volume (Resinomyces, Roridomyces) and 26 others not included (Arrhenia,

Baeospora, Blasiphalia, Chromosera, Chrysomphalina, Collybia, Contumyces,

Delicatula, Dendrocollybia, Fayodia, Gamundia, Gyroflexus/Sphagnomphalia,

Hemimycena, Hydropus, Hygroaster, Lichenomphalia, Loreleia, Marasmiellus,

Mycenella, Omphalina, Pseudobaeospora, Rickenella, Sphagnurus, Strobilurus,

and Xeromphalina).

Three dichotomous keys (all needed) are presented: [1] keys to mycenoid

genera & species (4 pages), [2] a macro-based key further divided into

Keys A-K (18 pages), and [3] a micro-based key (12 pages), where an entry

key allocating 5 species also provides leads to Keys I-III. Although I would

not have presented the spore Q >1.2 vs Q <1.2 as the first couplet in Key I,

Iam loath to criticize any key that has not been personally tested in the field and

lab. These dichotomous keys are augmented by an extremely helpful synoptic

key to mycenas sorted by habitat/substrate types.

The 115 species treatments comprising the bulk of the volume (pp. 78-333)

do not disappoint. The accepted taxonomic name and nomenclator (followed

by basionyms, synonyms, and misapplications) stand at the top of each first

page above a 2-3 line diagnosis. A complete technical description, ecology

& distribution data, and discussion follow. The text is accompanied by 2-5

excellent photos (most in situ, but often shown with a plate (taken with macro-

lens or dissecting microscope) detailing characters that might otherwise

be missed. Mycena mitis, a rare mushroom not captured fresh in the field, is

represented by a painting and line drawing. While the line drawings are rather

small and definitely not as detailed or elaborate as those found in Smith (1947),

Rexer (1994, who helpfully grouped elements), Maas G. (1992), and Robich

(2003, by far the most intricately rendered), those in Aronsen & Leessoe seem

somehow more accessible and less likely to confuse. Two species also offer a

table or list comparing two look-alikes (e.g., M. polygramma vs. M. vitilis, M.

filopes vs. M. metata), which I found particularly helpful.

Two new species, fully treated on pages 220-221 (M. pasvikensis) and 270-

271 (M. mucoroides), are also formally published on p. 334, accompanied by

the required MycoBank (& GenBank) numbers, etymology, diagnosis, and

type information. A note on the messy nomenclature surrounding M. rosea

nom. illeg. shows Bulliard’s plate used by Maas G. to lectotypify the species

with Bulliard as author. The taxonomic treatments conclude with comments

Book Reviews ... 731

on 47 taxa not treated in the main text, 11 rejected names, 19 dubious taxa, 4

extralimital taxa, 37 excluded taxa now transferred to other genera, and one

‘misinterpretation. References and a taxonomic index round out the volume.

Its stunning sharp yet luminous photos, uncluttered elegant design, and

wealth of information make THE GENUS MYCENA S.L. a joy to own, read, and

use. A reliable reference that is simultaneously striking and restful fosters

the illusion that the reader should easily be able to master the genus with no

problem—an illusion that will keep many of us soldiering on. I highly advise

everyone intrigued by these beautiful delicate agarics (and isn’t that everyone?)

to add this volume to their libraries. You won't regret it.

Breitenbach J, Kranzlin F. 1991. Fungi of Switzerland 3. Verlag Mycologia,

Lucerne. 361 pp., 349 col. ill.

Gminder A. 2016. Nomenclatural novelties. Index Fungorum 302: 1.

Kiihner R. 1938. Le genre Mycena. Encyclopédie mycologique X. P. Lechevalier,

Paris. 710 pp. (in French)

Maas Geesteranus, R.A. 1992. Mycenas of the northern hemisphere. I. Studies

in Mycenas and other papers. 391 pp.; II. Conspectus of the mycenas of

the northern hemisphere. 493 pp.; Koninklijke Nederlandse Akademie

van Wetenschappen verhandelingen, afd. Natuurkunde, Amsterdam.

Redhead SA. 2012. Nomenclatural novelties. Index Fungorum 14: 1.

Redhead SA. 2013. Nomenclatural novelties. Index Fungorum 15: 2.

Redhead SA. 2016a. Nomenclatural novelties. Index Fungorum 289: 1.

Redhead SA. 2016b. Nomenclatural novelties. Index Fungorum 290: 1

Redhead SA. 2016c. Nomenclatural novelties. Index Fungorum 291: 1

Redhead SA, Singer R. 1981. Resinomycena gen. nov. (Agaricales), an ally of

Hydropus, Mycena and Baeospora. Mycotaxon 13: 150-170.

Rexer K-H. 1994. Die Gattung Mycena s.]. Studien zu ihrer Anatomie,

Morphologie und Systematik. PhD dissertation, Fakultat fiir Biologie der

Eberhard-Karls- Universitat Tubingen. 318 pp. (in German)

Robich, Giovanni. 2003. Mycena d’Europa. A.M.B. Fondazione Centro Studi

Micologici, Trento, Italy. 728 p. (in Italian)

Smith, Alexander H. 1947. North American species of Mycena. University of

Michigan Press, Ann Arbor. 521 p.

MYCOTAXON

July-September 2016—Volume 131, pp. 733-734

http://dx.doi.org/10.5248/131.733

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 131(3)

Anaverticicladus P.M.O. Costa, Malosso & R.F. Castaheda [MB 816930], p. 688

Anaverticicladus uncinatus P.M.O. Costa, Malosso & R.E. Castaneda

[MB 816931], p. 688

Boletus durhamensis B. Ortiz, Bessette & McConnell [MB 810115], p. 706

Capsicispora J.Y. Wang, Yu M. Cai & X.G. Zhang [MB 81802], p. 560

Capsicispora mycophila J.Y. Wang, Yu M. Cai & X.G. Zhang [MB 81803], p. 560

Corynesporopsis obclavata Jian Ma [MB 816248], p. 583

Diaporthe henanensis Yi Yang, H.Y. Wu & Meng Zhang [MB 804356], p. 649

Diorygma fuscum Jian Li bis & Z.F. Jia [FN 570263], p. 718

Diploschistes tianshanensis A. Abbas, S.Y. Guo & Ababaikeli [MB 804328], p. 569

Ellisembia henanensis J.W. Xia & X.G. Zhang [MB 818289], p. 598

Erysiphe acantholimonis J.G. Song, B. Xu & H.D. Shin [MB 815918], p. 624

Gymnopus ramulicola T.H. Li & S.F Deng [MB 815128], p. 665

Harmoniella campanaensis Madrid, D. Torres & V. Silva [MB 815759], p. 536

Hemithecium hainanense Z.F. Jia [FN 570071], p. 672

Laboulbenia camerunensis T.W. Wang & Haelew. [MB 811237], p. 615

Lithothelium bermudense F. Berger, LaGreca & Aptroot [MB 815588], p. 528

Marthamyces coronadoae Raymundo, R. Valenz. & Esqueda [MB 815275], p. 522

Periconia notabilis Chuaseehar., Somrith. & Boonyuen [MB 815470], p. 503

Phaeomonilia nanningensis C.L. Yang & X.G. Zhang [MB 818085], p. 548

Pseudoacrodictys ambigua J.M. Gao & X.G. Zhang [MB 818098], p. 559

Puccinia tatarinovii Kom. & Tranzschel 1939 (lectotypified & epitypified), p. 657

Rosellinia hainanensis Wei Li bis & L. Guo [FN 570257], p. 542

Sporidesmiella jiulianshanensis Jian Ma [MB 816247], p. 578