IV ... MYCOTAXON 132(1)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-TWO (1) — TABLE OF CONTENTS

132-1: TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS & EDITORIALS

Nomenclatural novelties & typifications ........ 0. cece eee ee eee ees vii

CVICWE RO AS SEN, 6 Rae Le eee te Se Oe eee PCE URE She Viii

ESOP IPGIE TE ANON a Sa Salle Sane MaMa ag atte he ia mt ease teh ly eRe ps ix

LOL SA DIISSION PT OCR AGT * FAS xeon eet nee Sete RE Re ae ep ests te Seed xi

RESEARCH ARTICLES

A new host for Puccinia menthae

SANLI KABAKTEPE, TURAN ARABACI & TURGAY KOLAC

Mycelephas levisporus sp. nov. on submerged wood

from a freshwater habitat in Brazil Taimy CANTILLO-PEREZ,

JULIO MENA-PORTALES & Luis FERNANDO PASCHOLATI GUSMAO

Scutellospora tepuiensis sp. nov. from the highland tepuis

of Venezuela Z1TA DE ANDRADE',

EDUARDO FURRAZOLA & GISELA CUENCA

Five penzigioid Xylaria species from Veracruz (Mexico)

FIDEL TaptA, YU-MING Ju, SANTIAGO CHACON & ELSA UTRERA-BARILLAS

Three species of wood-decaying fungi in Polyporales

new to China CHANG-LIN ZHAO, SHI-LANG LIv,

GUANG-JUAN REN, XIAO-HONG JI & SHUANGHUI HE

Circinella (Mucorales, Mucoromycotina) from China

RU-YONG ZHENG, XIAO-YONG LIU & YA-NING WANG

Astraeus ryoocheoninii sp. nov. from Korea and Japan

and phylogenetic relationships within Astraeus

RuIM Ryoo, HonG-Duck Sou,

KANG-HYEON Ka & HYUN PARK

New records of Caloplaca, Hydropunctaria, and Verrucaria

from Turkey and Asia KapDIR KINALIOGLU

Chloridium terricola sp. nov. from China YONG WANG, CHUN-YU JIE,

KEVIN D. HybDE, Yu-LAN JIANG,

TIAN-YU ZHANG, & DE-GANG ZHAO

Erysiphe russellii confirmed in Korea by morphological

and sequence analyses Tui THuoNG THUONG NGUYEN & HyANG-BuRM LEE

Phlebia brevibasidia sp. nov. from India

GURPREET KAurR, AVNEET P. SINGH & G.S. DHINGRA

JANUARY-MARCH 2017... V

Two new records of puffballs in Thailand

JATURONG KUMLA, NAKARIN SUWANNARACH & SAISAMORN LUMYONG 99

Cyanodermella asteris sp. nov. (Ostropales)

from the inflorescence axis of Aster tataricus LINDA JAHN,

THOMAS SCHAFHAUSER, STEFAN PAN, TILMANN WEBER,

WOLFGANG WOHLLEBEN, DAVID FEWER, KAARINA SIVONEN,

LIANE FLOR, KARL-HEINZ VAN PEE, THIBAULT CARADEC,

PHILIPPE JACQUES, MIEKE M.E. HUIJBERS,

WILLEM J.H. VAN BERKEL & JUTTA LUDWIG-MULLER 107

Eight Caloplaca species newly recorded from Bolivia,

including C. crocina comb. nov. KaRINA WILK & ADAM Fiaxus 125

Notes on rust fungi in China 3.

Puccinia adenocauli comb. nov. and its life cycle and new host

JING-XIN JI, QI WANG, ZHUANG LI, Yu LI & MAKOTO KAKISHIMA 141

Colletotrichum fioriniae comb. & stat. nov.,

resolving a nomenclatural muddle SHAUN R. PENNycooK 149

Records of terricolous lichens from Ecuador YADIRA GONZALEZ,

GREGORIO ARAGON, ANA Rosa BuRGAZ & Maria PrrETO 153

Three lichen species in Buellia, Catillaria & Cheiromycina,

new to Poland MartIN Kuxwa, PawEzt CZARNOTA & ANNA LUBEK 177

Additions to the mycobiota of Poland ANNA LUBEK & MarTIN Kuxwa 183

Macrocyclic Edythea quitensis rust on Berberis hallii

in Ecuador Marta E. ORDONEZ & CHARLES W. BARNES 197

Leucoagaricus ariminensis sp. nov., a lilac species

from Italy F. Dovana, M. Contu, P. ANGELI, A. BRANDI & M. MucctarELLi 205

Geastrum reinkingii reconsidered

LARISSA TRIERVEILER-PEREIRA & ROSA MARA BORGES DA SILVEIRA 217

Bacidia, Micarea, Sagedia, and Stigmidium spp.

new to Turkey KADIR KINALIOGLU & ANDRE APTROOT 223

Urocystis narcissi, a new record from Asia

SADIQULLAH, A. IsHAQ, M. Fiaz, A.N. KHALID & H. AHMAD 231

Lembosia dianesei sp. nov. associated with Peritassa campestris

in Minas Gerais, Brazil MARUZANETE PEREIRA MELO,

JosE Luiz BEZERRA, SILVINO INTRA MOREIRA & EDUARDO ALVES 235

REGIONAL MYCOBIOTAS NEW TO THE MYCOTAXON WEBSITE 241

A checklist of clavarioid fungi (Agaricomycetes) recorded in Brazil

— ANGELINA DE MEIRAS-OTTONI, LIDIA SILVA ARAUJO-NETA

& TATIANA BAPTISTA GIBERTONI

vI ... MYCOTAXON 132(1)

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-ONE (4)

MYCOTAXON for OCTOBER—DECEMBER 2016, (I-xII + 735-1000)

was issued on January 30, 2017

JANUARY-MARCH 2017...

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 132(1)

Astraeus ryoocheoninii Ryoo

[MB 804156], p. 68

Caloplaca crocina (Kremp.) K. Wilk & R. Vargas

[MB 816545], p. 127

Chloridium terricola Yong Wang bis, Jie & K.D. Hyde

[MB 820926], p. 80

Circinella nodulosa R.Y. Zheng, X.Y. Liu & Y.N. Wang

[FN 570151], p. 54

Circinella ramosa R.Y. Zheng, X.Y. Liu & Y.N. Wang

[FN 570150], p. 56

Colletotrichum fioriniae (Marcelino & Gouli) Pennycook

[IF 553097], p. 150

Cyanodermella asteris L. Jahn & Ludw.-Mill.

[MB 814158], p. 113

Lembosia dianesei M.P. Melo, J.L. Bezerra, S.I. Moreira & E. Alves

[MB 814528], p. 236

Leucoagaricus ariminensis Dovana, Angeli, Contu & Brandi

[IF 552049], p. 209

Mycelephas levisporus Cantillo, J. Mena & Gusmao

[MB 815045], p. 6

Phlebia brevibasidia G. Kaur, Avn.P. Singh & Dhingra

[MB 816998], p. 95

Puccinia adenocauli (Syd. & P. Syd.) Jing X. Ji & Kakish.

[MB 817618] (neotypified, epitypified), p. 144

Scutellospora tepuiensis Furrazola & Cuenca

[MB 818417], p. 11

VII

Vul ... MYCOTAXON 132(1)

REVIEWERS — VOLUME ONE HUNDRED THIRTY-TWO (1)

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

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

prepared for this quarter.

Najam-ul-Sehar Afshan

Teuvo Ahti

M. Catherine Aime

Gerald L. Benny

Janusz Blaszkowski

Tiara S. Cabral

Rafael F. Castafieda-Ruiz

Pablo Pérez Daniéls

Cvetomir M. Denchev

José Carmine Dianese

Zai-Wei Ge

Nils Hallenberg

Maria L. Hernandez Caffot

Mikael Jeppson

Peter Johnston

Sevda Kirbag

Paul M. Kirk

Irmgard Krisai-Greilhuber

Scott LaGreca

Fidel Landeros

De-Wei Li

Laszl6 L6k6s

Robert Liicking

Tom May

Eric H.C. McKenzie

David J. McLaughlin

Jurga Motiejunaite

Gregory M. Mueller

Lorelei L. Norvell

Shaun R. Pennycook

Jadergudson Pereira

Olinto Liparini Pereira

Ronald H. Petersen

Raquel Pino-Bodas

Michele Piercey-Normore

Huzefa A. Raja

Andrea I. Romero

Mark R.D. Seaward

Hacer Sert

B.M. Sharma

H.J.M. Sipman

Steven L. Stephenson

Jeffrey Stone

Sidney Luiz Stiirmer

Reinaldo Vargas Castillo

Josef Vlasak

Felipe Wartchow

Andrew W. Wilson

Li-Wei Zhou

JANUARY-MARCH 2017... IX

FROM THE EDITOR-IN-CHIEF

RICHARD P. KorF IN FULL COLOR—Those wishing a full color printed version of our

memoriam to MycotTaxon’s co-founder may order a special copy of “Richard P. Korf

(1925-2016): A Celebration” for $8 on the MycoTAXON website.

Surviving Co-founder Grégoire Hennebert recently corrected the 1973 date under

the photo of the two founders in Louvain, noting that Dick first visited him in Belgium

in 1972 on his Fulbright Fellowship. During Dick’s stay at the Laboratory of Systematic

Mycology at Louvain University where he worked with Grégoire on Botrytis and

conidial states of Sclerotiniaceae (see Hennebert, “Botrytis and Botrytis-like genera,

PERSOONIA 1973), Grégoire expressed a desire to launch a new mycological journal

(named ‘Biourgia’ for the Belgian Penicillium specialist, Prof. Biourge) “because I

had lost priority of one new species due to slow publication in a traditional scientific

journal”

The two colleagues later discussed setting up a new journal during Grégoire'’s

Sep. 23-30 1973 visit with Dick at Cornell in Ithaca and his Nov. 5-Dec. 14 1973

stint as a Visiting Professor in Canada and the United States. Feeling that the editorial

work might be too time-consuming for Grégoire (who had “no hope for assistance

in Louvain’), Dick suggested serving as Co-editor. He had previously published

a few papers in PHYTOLOGIA, an offset botanical publication and felt that editing

ready-to-print manuscripts would be a time-saving solution. Dick proposed a more

international title, MycoTaxon, for their journal and that he manage the edition from

Ithaca, with Grégoire agreeing to control “the book review job.’

At the time Grégoire still had no electric typewriter in Louvain, and so manually

prepared his paper (with fully justified lines) on Lomentospora prolifica, a new

hyphomycete, for MycoTaxon’s first issue (1974, MycoTaxon 1(1): 45-50), which

was, according to Grégoire, “a real hard job, a problem soon solved by the computer.”

[Younger readers may not realize how much time and paper it took to line up each

right-hand margin, involving multiple attempts at spacing words with the “/” key at the

margin before inserting interior spaces in the old Courier font where every letter and

space had the same width.] Many thanks to Grégoire for these ‘ancient’ historical notes!

BREAK WITH TRADITION—Since its inception, MycoTaxon has presented its

‘nomenclatural novelties’ at the close of each volume (1974-2014 p.p.) or issue (2014

ff.). Our ‘nom novs’ section (which lists new combinations as well as names new to

science and expanded to incorporate typifications in 2012) has until now appeared

online as a separate free-access PDE However, it makes more economic sense to

absorb the novelties within the separate cover section (also free access). To aid

those who will—no doubt—continue to search the end of each issue for the ‘new

nomenclature, we now entitle our cover sections with the issue number followed by

“TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS & EDITORIALS.” This issue's

newly proposed names (with authorities, nomenclatural identification numbers, and

page where published) are listed on p. vii of this free access 132(1) cover section.

NEW REQUIREMENTS FOR RANGE EXTENSION PAPERS—Your editors have fretted

for some time over problems presented by papers reporting on species outside of

x ... MYCOTAXON 132(1)

formerly published distribution ranges. One problem has been the presentation of

capsule ‘field-guide type’ descriptions that do not describe the specimens in sufficient

detail to demonstrate that they represent the species to which they are referred.

In keeping with our taxonomic focus, we alert authors to the necessity of devoting

as much care to range extension descriptions as they do for a new species and ask

them to point out differences that might indicate the existence of a cryptic taxon

warranting research by an expert.

To that end, we have developed the following requirements for authors intending

to publish a previously described taxon in MyCOTAXON.

An acceptable taxonomic contribution must include an original description and

illustration(s) of the newly collected specimens. Additionally, for each taxon range

extension, authors must include:

[1] a TAXONOMIC HEADING including the authorship, bibliographic reference, and

the figure references;

[2] an ORIGINAL technical description based ENTIRELY on the newly collected

specimens (no information from previously published descriptions should be included

in this first description);

[3] a standard SPECIMENS EXAMINED section, listing the collection and vouchering

data for the newly collected specimens; and

[4] a brief piscussion of similarities/differences between the newly collected

specimens and previously published descriptions and data from the same species.

2017 GUIDES & FORMS—Prospective authors should obtain the 2017 guide, sample

manuscript, blank template and peer reviewer and submissions forms from

Mycotaxon’s Author Download page on <www.mycotaxon.com>. Note: there are

two DIFFERENT Sets of forms: one for papers to be published in the journal (requiring

two expert reviewers) and a second set for annotated species lists (mycobiotas) to be

posted on our Regional Checklist Page (requiring three expert reviewers).

MyYCOTAXON 132(1) contains 26 research papers by 94 authors (representing 23

countries) and revised by 49 expert reviewers.

Within its pages are 10 species new to science [representing Astraeus from

Korea; Chloridium and Circinella from China; Cyanodermella from an Austrian

cultivar; Lembosia and Mycelephas from Brazil; Leucoagaricus from Italy; Phlebia

from India; and Scutellospora from Venezuela] and new combinations in Caloplaca,

Colletotrichum, and Puccinia.

In addition to range extensions [lichens in Bolivia, Poland, and Turkey; polypores

in China; powdery mildew in Korea; rust in Pakistan; puffballs in Thailand] and a new

host for Puccinia menthae in Turkey, we also offer the complete life cycle for a rust

in Ecuador, excellent coverage of five penzigioid Xylaria species in Vera Cruz, and

molecular confirmation of Geastrum reinkingii as an independent species.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

21 April 2017

JANUARY-MARCH 2017... XI

2017 MyYCOTAXON SUBMISSION PROCEDURE

Prospective MycoTaxon authors should download the MycoTaxon 2017 guide, review

& submission forms, and MycoTaxon sample manuscript by clicking the ‘file download

page’ link on our INsTRUCTIONS TO AUTHORS page before preparing their manuscript.

This page summarizes our “4-step’ submission process.

1—PEER REVIEW: Authors first contact two (for journal papers) or three (for

annotated species ‘weblists’) peer reviewers before sending them formatted text &

illustration files and the appropriate 2017 MycoTaxon journal or weblist reviewer

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4—FINAL EDITORIAL REVIEW: Files with errors will be rejected or returned for

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The Mycotaxon Webmaster <mycotaxon@gmail.com> posts announcements,

subscription & publications information, and author forms & templates on the

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

subscription articles are offered.

MY COTAXON

January-March 2017— Volume 132, pp. 1-3

http://dx.doi.org/10.5248/132.1

A new host for Puccinia menthae

SANLI KABAKTEPE ', TURAN ARABACI’ & TuRGAY KOLac?

' Battalgazi Vocational School, Inonu University, TR-44210 Battalgazi, Malatya, Turkey

? Department of Pharmaceutical Botany, Faculty of Pharmacy, Inonu University, Malatya, Turkey

° Vocational School of Health, Inonu University, Malatya, Turkey

* CORRESPONDENCE TO: sanli.kabaktepe@inonu.edu.tr

ABSTRACT—Cyclotrichium (Lamiaceae) is reported as a new host genus for the rust fungus,

Puccinia menthae. The morphological and microscopical characteristics of this fungus are

described and illustrated based on the collected materials.

Key worps—Malatya, Pucciniales, taxonomy, Turkey

Introduction

The genus Cyclotrichium (Boiss.) Maden. & Scheng. (Lamiaceae, tribe

Mentheae) contains nine species distributed in Turkey, Lebanon, Iraq, and Iran.

Six Cyclotrichium species have been reported in Turkey, including two endemic

species: C. glabrescens (Boiss. ex Rech.f.) Leblebici and C. niveum (Dirmenci et

al. 2010, Dirmenci 2012). To our knowledge no rust fungi have previously been

described on Cyclotrichium or cited for earlier synonyms of the species.

Puccinia menthae is an autoecious, macrocyclic, long-cycle rust recorded on

28 genera of Lamiaceae (Farr & Rossman 2015).

In Turkey, Puccinia menthae has been reported on Calamintha, Clinopodium,

Mentha, Micromeria, Origanum, and Satureja (Bahcecioglu & Kabaktepe 2012).

Here we present a record of Puccinia menthae on a new host genus,

Cyclotrichium, from Malatya province, Turkey.

Materials & methods

The material on which this study is based was collected from Malatya in 2015. The

host specimen was prepared according to established herbarium techniques. The host

plant was identified according to Leblebici (1982) and Dirmenci et al. (2010). Spores

2 ... Kabaktepe, Arabaci & Kola¢

“ae . : eu. .

Fic. 1. Puccinia menthae on Cyclotrichium niveum. A, urediniospores; B, teliospores; C, infected

brown areas on leaf tissue; D, herbarium specimen (INU 1200).

Cyclotrichium new host for Puccinia menthae (Turkey) ... 3

were scraped from the dried host specimen, mounted in lactophenol, and measured

using Analysis LS Starter software.

Names and families of plants are given according to http://www.theplantlist.org. The

specimen is preserved in the Inonu University Herbarium, Battalgazi, Malatya, Turkey

(INU).

Taxonomy

Puccinia menthae Pers., Syn. Meth. Fung.: 227 (1801). Fia. 1

Spermogonia and aecia not found. Uredinia hypophyllous, rounded,

scattered, 0.5-1 mm, surrounded by the epidermis, sometimes confluent,

yellowish or brownish. Urediniospores globose to obovoid or ellipsoid, 17-28 x

14-19 um; wall <2.5 um thick, echinulate, hyaline to light yellow; germ pores 2,

equatorial. Telia mostly hypophyllous, rarely amphigenous, on leaves, petioles

and stems, rounded, blackish brown, scattered, pulverulent, 0.5-1.5 mm.

Teliospores ellipsoid to obovoid, rounded at both ends, not or only slightly

constricted at the septum, 26-35 x 19-23 um, verrucose (sometimes smooth),

brown, wall 1.5-3 um thick at sides, 3-5 um thick apically, apical in upper

cell and close to septum in lower cell, with a hyaline papilla; pedicel slender,

hyaline, longer than the spores.

SPECIMEN EXAMINED—On Cyclotrichium niveum (Boiss.) Manden. & Scheng.

(Lamiaceae). Turkey: Malatya, Konak, Beydagi, 1700-1750 m a.s.l., 6.Sep.2015, T. Kola¢

1054 (INU 1200).

REMARKS: Our record of Puccinia menthae on Cyclotrichium niveum increases

the host list of this cosmopolitan rust species to include 29 lamiaceous genera.

Acknowledgments

We thank Prof. Dr. Sevda Kirbag and Prof. Dr. Hacer Bakir Sert for presubmission

expert reviews.

Literature cited

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

494. http://dx.doi.org/10.5248/119.493].

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

Dirmenci T. 2012. Cyclotrichium (Boiss.) Manden. & Scheng. p. 553, in: A Giiner et al. (eds).

A Checklist of the Flora of Turkey (Vascular Plants). Istanbul: Nezahat Gékyigit Botanik

Bahcesi ve Flora Arastirmalari Dernegi Yayini (In Turkish).

Dirmenci T, Diindar E, Deniz G, Arabaci T, Martin E, Jamzad Z. 2010. Morphological, karyological

and phylogenetic evaluation of Cyclotrichium: a piece in the tribe Mentheae puzzle. Turkish

Journal of Botany 34: 159-17.

Farr DF, Rossman AY. 2015. Fungal Databases, Systematic Mycology and Microbiology Laboratory,

ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/ [Accessed November 24, 2016]

Leblebici E. 1982. Cyclotrichium (Boiss.) Manden. & Scheng. 346-349, in: PH Davis (ed.). Flora of

Turkey and the East Aegean Islands, Vol. 7. Edinburgh, Edinburgh University Press.

MY COTAXON

January-March 2017— Volume 132, pp. 5-8

http://dx.doi.org/10.5248/132.5

Mycelephas levisporus sp. nov. on submerged wood

from a freshwater habitat in Brazil

TAIMY CANTILLO-PEREZ’, JULIO MENA-PORTALES?

& Luis FERNANDO PASCHOLATI GUSMAO™

' Lab. de Micologia, Dpto. de Ciéncias Biologicas, Universidade Estadual de Feira de Santana,

Avenida Transnordestina s/n, Bairro Novo Horizonte, 44036-900, Feira de Santana, Brazil

? Instituto de Ecologia y Sistematica,

Carretera Varona 11835 e/ Oriente y Lindero, CP 11900, Boyeros, La Habana, Cuba

*CORRESPONDENCE TO: lgusmao@uefs. br

ABSTRACT—A new species from submerged wood in Brazil, Mycelephas levisporus, is

described and illustrated. The long conidiophores and smooth wide conidia with slightly

constricted septa are the main characteristics that distinguish the new species from the type

species.

Key worps—hyphomycetes, aeroaquatic fungi, helicoidal, neotropic

Introduction

During a study of microfungi occurring on submerged plant debris in the

semi-arid Northeast region of Brazil, an interesting species of Mycelephas

R.F. Castaneda was collected on submerged wood.

Mycelephas (Gams et al. 2009) is a monotypic genus described from dead

wood in Cuba (Castafieda-Ruiz 1984; as Arnoldiella nom illeg.); it differs from

other helicoidal microfungi by bifurcate, almost cochleate conidia coiled in

two different planes (Castafeda-Ruiz 1984; Goos 1987; Seifert et al. 2011).

Despite these unique morphological characters, other species such as Helicoma

resinae (Lindau) J.L. Crane & Schokn, which has mono- or polyblastic

conidiogenous cells, could be confused with Mycelephas based on its conidia

coiled 3-4.5 times in one plane. Diplorhynchus G. Arnaud (Arnaud 1952)

and Dichotomophthoropsis M.B. Ellis (Ellis 1971) also share some similarities

6 ... Cantillo-Pérez, Mena-Portales & Gusmao

with Mycelephas. Diplorhynchus has hyaline conidia coiled in one plane and

bifurcated at the end, and due to these characters Goos (1987) considered that

genus as congeneric with Mycelephas; as no other records for Diplorhynchus

have been registered since Arnaud (1952), however, its taxonomic status

remains uncertain. Dichotomophthoropsis can be distinguished by its tretic

conidiogenous cells, lobed cells, and helicoid conidia that do not bifurcate

(Ellis 1971).

Materials & methods

Samples of submerged, decayed wood were collected in streams of Cocho River in

Piata and placed in plastic bags. In the laboratory, wood pieces were washed and placed

in moist chambers. Subsequently, the moist chambers were placed in a large polystyrene

box with sterile water plus glycerol at 24°C for 30 days and were examined periodically

or once a week (Castafieda-Ruiz 2005). Permanent slides were prepared in PVL and

micrographs were obtained with an Olympus microscope BX 51 with bright field and

Nomarski interference optics. The type specimen of the new species is deposited in the

Herbarium of Universidade Estadual de Feira de Santana, Bahia, Brazil (HUEFS).

Taxonomy

Mycelephas levisporus Cantillo, J. Mena & Gusmao sp. nov. Fia. 1

MycoBAnk MB 815045

Differs from Mycelephas robustus by its longer conidiophores and smooth and wider

conidia.

Type: Brazil. Bahia: Piata, Chapada Diamantina, Coché River, 13°00’S 41°51’W, on

submerged wood. 3.IV.2014, coll. T. Cantillo (Holotype, HUEFS 216594; isotype,

HUEFS 216641).

Erymo oey: Latin, levis + sporus,-referring to the smooth conidia surface.

ANAMORPHIC FUNGI, HYPHOMYCETES. COLONIES on natural substrate pulvinate,

granulate, yellow to yellowish brown, aggregated. MycELIUM mostly immersed

in the substrate, septate, branched, brown. CONIDIOPHORES solitary, simple or

branched, straight to flexuous, septate, smooth, light brown, turning brown

with age, 93-205 x 7.5-15 um; 7.5-12.5 um at the apex. CONIDIOGENOUS CELLS

mono or polyblastic, integrated, terminal or intercalary, sympodial, cylindrical,

with dark scars. CONIDIAL SECESSION schizolytic. CONIDIA solitary, dry, light

brown to subhyaline, smooth, helicoidal, 6-8 septa in the main filament,

slightly constricted at septa. The filament bifurcates in two branches coiled

2-3 times giving an almost cochleate appearance, 9-15 um diam. Mature

conidia 23-(38)42.5 um in side view, 25-37.5 um in frontal view. The basal

cells of conidia are V-shaped with truncate base; sometimes darker than other

cells, 8-15 um long, 2-3 um at the base, and 6-10 um at the apex.

Mycelephas levisporus sp. nov. (Brazil) ... 7

Fic. 1. Mycelephas levisporus (HUEFS 216594, holotype). A: immature conidium in side

view; B-F: conidia in frontal view showing bifurcation of filament in two branches in

different coiling stages; G: fully developed conidium; H: conidiophores and conidia;

I: detail of conidiogenous cell; J, K: conidiophores showing integrated conidiogenous cells

with dark scars. Scale bars: A-H, J = 10 um; I = 2 um; K = 20 um.

OTHER MATERIAL EXAMINED: Mycelephas robustus (R.F. Castafieda) R.E. Castafieda.

Cuba. La Habana: Santiago de Las Vegas, on dead branches of Pseudolmedia spuria (Sw.)

Griseb., 1.[II.1983, coll. R.F. Castafeda (Holotype, C83/39 INIFAT).

Notes: Mycelephas robustus (R.E. Castafieda) R.F. Castafeda differs from

M. levisporus by its smaller conidiophores (41-80 um long) and its smaller

rugose conidia (20-27 um diam.; Castafieda-Ruiz 1984, as Arnoldiella robusta).

Acknowledgments

The authors express their sincere gratitude to Dr. Huzefa Raja and Dr. José C. Dianese

for their critical review of the manuscript and Dr. Rafael Castafieda for comments. We

also thank the Programa de Pés-Graduacao em Botanica (PPGBot/ UEFS) and Program

8 ... Cantillo-Pérez, Mena-Portales & Gusmao

of Research on Biodiversity in the Brazilian Semi-arid (PPBIO Semiarid - MCTI/CNPgq,

proc. 457498/2012-9) for financial support. The first author thanks PEC-PG/CAPES

(Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior/ proc. 12636134) for

scholarships, and LFPG also thanks CNPq (proc. 305413/2011-2) for grant.

Literature cited

Arnaud G. 1952. Mycologie concréte: genera. Bulletin de la Société Mycologique de France 68:

19-33;

Castaneda-Ruiz RF. 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.

Castafeda-Ruiz RF. 2005. Metodologia en el estudio de los hongos anamorfos. 182-183, in:

Anais do V Congresso Latino Americano de Micologia. Brasilia.

Ellis MB. 1971. Dematiaceous Hyphomycetes. X. Mycological Papers 125. 30 p.

Gams W, Seifert KA, Morgan-Jones G. 2009. New and validated hyphomycete taxa to resolve

nomenclatural and taxonomic issues. Mycotaxon 110: 89-108. https://doi.org/10.5248/110.89

Goos RD. 1987. Fungi with a twist: the helicosporous hyphomycetes. Mycologia 79: 1-22.

https://doi.org/10.2307/3807740

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

Biodiversity Series 9. 997 p.

MY COTAXON

January-March 2017— Volume 132, pp. 9-18

http://dx.doi.org/10.5248/132.9

Scutellospora tepuiensis sp. nov.

from the highland tepuis of Venezuela

ZITA DE ANDRADE't, EDUARDO FURRAZOLA’” & GISELA CUENCA'™*

' Instituto Venezolano de Investigaciones Cientificas (IVIC), Centro de Ecologia,

Apartado 20632, Caracas 1020-A, Venezuela

?Instituto de Ecologia y Sistemdtica, CITMA,

C.P. 11900, Capdevila, Boyeros, La Habana, Cuba

* CORRESPONDENCE TO: gcuenca@ivic.gob. ve

ABSTRACT—Examination of soil samples collected from the summit of Sororopan-tepui at

La Gran Sabana, Venezuela, revealed an undescribed species of Scutellospora whose spores

have an unusual and very complex ornamentation. The new species, named Scutellospora

tepuiensis, is the fourth ornamented Scutellospora species described from La Gran Sabana and

represents the first report of a glomeromycotan fungus for highland tepuis in the Venezuelan

Guayana.

Key worps—arbuscular mycorrhizal fungus, taxonomy, tropical species, Gigasporaceae,

Glomeromycetes

Introduction

The Guayana shield occupies a vast area that extends approximately 1500

km in an east-to-west direction from the coast of Suriname to southwestern

Venezuela and adjacent Colombia in South America. This region is mostly

characterized by nutrient-poor soils and a flora of notable species richness,

high endemism, and diversity of growth forms (Huber 1995). Vast expanses of

hard rock (Precambrian quartzite and sandstone) that once covered this area as

part of Gondwanaland (Schubert & Huber 1989) have been heavily weathered

and fragmented by over a billion years of erosion cycles, leaving behind just

a few strikingly isolated mountains (Huber 1995). These table mountains,

with their sheer vertical walls and mostly flat summits, are the outstanding

physiographic feature of the Venezuelan Guayana (Schubert & Huber 1989).

The Pemon Amerindians of southeastern Venezuela call these mountains

10 ... De Andradet, Furrazola & Cuenca

“tepuis” (Huber 1995). A tepui is a sandstone mountain or tableland with a

flat summit and vertical walls up to 600-3000 m tall. Rising abruptly over

the forests and savannas of La Gran Sabana, Venezuela, these tepuis are quite

isolated by their peculiar shape and harbor a high degree of endemic flora and

fauna. Soils of tepuis are acid and extremely oligotrophic compared with their

adjacent lowlands.

The often-steep tepui slopes are covered with dense mossy forests bathed

in moisture from dense clouds that form along the cliffs. Many tepui summits

have relatively little soil that is exposed to temperatures averaging 18-24 °C and

heavy rainfall above 2000 mm. Dense clouds and prolonged mist are common

and provide additional moisture at highland sites.

Arbuscular mycorrhizal fungi (AMF) are widespread in most terrestrial

ecosystems where they form mutualistic associations with most plants and

facilitate nutrient uptake from the soil via an extensive extraradical mycelium

(Smith & Read 2008). This mycorrhizal association, which has a low degree of

specificity between partners, involves a plant host and a fungus of the phylum

Glomeromycota (Brundrett 2009, Smith et al. 2011). Although the fungi

contribute many effects that may increase host plant survival and fitness, the

presumed primary benefit to the plant is improved nutrition in exchange for

carbohydrates to the fungal partner (Corréa et al. 2014).

Previous works have reported an important role for glomeromycotan fungi

in all lowland ecosystems of Venezuelan Guayana (Cuenca et al. 2003a,b),

and three Scutellospora species have been described from the lowlands:

S. spinosissima (Walker et al. 1998), S. crenulata (Herrera-Peraza et al. 2001),

and S. striata (Cuenca & Herrera-Peraza 2008). However, the presence of this

fungal group on tepui summits has remained unexplored.

During two brief trips to the Sororopan-tepui in Venezuelan Guayana, we

found scutellosporoid spores with a very complex ornamentation representing

an undescribed species. The fungus, described here as Scutellospora tepuiensis,

constitutes the first report of an arbuscular mycorrhizal fungus for the highland

tepuis.

Materials & methods

The 2050 m high Sororopan-tepui is part of the Ptari massif, a small mountain system

lying north of La Gran Sabana at 5°40-50’N 61°40-50’W, where annual precipitation

exceeds 2000 mm (Huber 1995). During two short trips made by helicopter to the

summit of Sororopan-tepui, composite soil samples (15 subsamples taken 0-15 cm

below ground level) were collected in two different tepui vegetation zones. Zone 1

was characterized by a low shrubland growing on acidic (pH 5.7), relatively deep soils

rich (20%) in organic matter; the dominant vegetation (<6 m tall) comprised woody

species in the Clusiaceae and Melastomataceae. Zone 2 was characterized by pioneer

vegetation growing between fissures and depressions of sandstone on shallow soil with

Scutellospora tepuiensis sp. nov. (Venezuela) ... 11

dominant species representing Poaceae, Cyperaceae, Melastomataceae, and Orchidaceae;

zone 2 soils are also acidic (pH 5.9) but with less (16.5%) organic matter than in zone

1. We found our new Scutellospora species in both vegetation types accompanied by

Scutellospora spinosissima C. Walker & Cuenca, Acaulospora morrowiae Spain &

N.C. Schenck, several Glomus species, and another undescribed Scutellospora.

Open pot trap cultures started from the soil samples were maintained in a glasshouse

for three months and then dried for one week. Two pre-germinated Vigna luteola seeds

(the host plant) were placed in 1-liter pot with undiluted field soil; the pot contents were

stored at room temperature (~20°C) for almost one year to break spore latency (Morton

et al. 1993), after which the spores were isolated and used to start pure cultures. Because

these last pure cultures were unsuccessful, the description of the species presented below

is based only on spores isolated from trap cultures (IVIC-38) and field soils.

Spores were isolated from the trap pots or field soils by wet sieving, decanting,

and sucrose centrifugation (Sieverding 1991). The isolated spores were suspended in

water and illuminated with light from a quartz-iodine fibre-optic source. Their color

was determined by comparison with a British color chart for fungi (Anon 1969). The

specimens were mounted in polyvinyl alcohol lacto-glycerol (PVLG) or in PVLG mixed

with Melzer’s reagent (1:1, v/v). Wall description and terminology follow Walker (1983)

and Walker & Vestberg (1998). Type material was deposited in the Venezuelan National

Herbarium, Universidad Central de Venezuela, Caracas, Venezuela (VEN) and the

Cuban National Herbarium, IES-CITMA, La Habana, Cuba (HAC).

Spore wall ornamentation and structure was also examined with scanning electron

microscopy (SEM). After removing or crushing the outermost spore wall layer with

fine tweezers under a dissecting microscope, we rinsed the spores in a phosphate buffer

solution prior to fixing in 1% osmium tetroxide at 4 °C for 1 h. Fixed samples were

dehydrated for 5 min in each dilution of an acetone series (20, 40, 70, 80, and 100%),

dried at critical point with liquid CO,, placed on aluminum metal holders, coated with

200 A gold-palladium, and then carefully examined by SEM.

Taxonomy

Scutellospora tepuiensis Furrazola & Cuenca, sp. nov. Figs 1-3

MycoBank MB 818417

Differs from all other Scutellospora spp. by its spores formed singly in soil and by the

complex spore ornamentation comprising numerous annulate pores, each surrounded

by a lip composed of either gemmae or columellae.

Type: Venezuela, La Gran Sabana Region, Sororopan tepui; isolated from the

rhizospheric soil of sclerophyllous shrubland, and pioneer vegetation growing between

fissures and depressions of sandstones on a shallow soil under Poaceae and Cyperaceae,

isolation date December 1996 by Z. De Andrade (Holotype: VEN-438658 [De Andrade

114-slide 2]; isotype: HAC-G-VE01 [De Andrade 114-slide 5).

EryMo_oey: Latin tepuiensis, referring to the “tepui” tableland where the species was

found.

Spores formed singly in soil terminally (rarely laterally) on a bulbous base

(sporophore); saffron (49) to orange (48) when young, becoming sienna (11)

12 ... De Andradet, Furrazola & Cuenca

to fulvous (12) when mature; generally globose or subglobose, 162-216 x

159-219 um (mean 191 x 190 um, n = 28), and containing oily globular drops

8-35 um diam.

SPORE WALL arrangement A(EL.M)B(MMM) or A(EL,)B(M)C(MMm) (following

Walker 1983), here referred to A(12,3)B(456) or A(12.,)B(3)c(456), with layer 2.

comprising two sub-layers: 2,4 and 2 B.

OUTERMOST LAYER (1) rarely observed; when present, hyaline, smooth,

<1 um thick, readily visible in younger spores but frequently detaching in older

ones; considered evanescent as not commonly found in mature spores.

LAYER 2, 7-10 um thick, laminated, composed of two sub-layers (2,4, 2.B).

SUB-LAYER 2.4 dark yellow, 5-7 um thick, perforated on its upper surface

with 1-5 um rounded to irregular pores, further inward apparently annulated

and completely filled by round to elliptical or irregular 1.5-7 um wide

annuli separated by 1-7 um with the in-between surface slightly depressed;

each annulus with ~1-um thick lip surrounded by 10-20 rounded gemmae

or elliptical to clavate columellae, 1-3 um long and ~1 um thick, extending

radially around the annulus pore as though hanging down into the lips (pores

and annuli best seen in surface view); inward from each gemma or columella

tip arise yeast-like projections, seemingly composed of chains or clusters

of tiny (much smaller than 0.5 um) wall material granules, the chains and

clusters brush-like to diffuse and lacking in structure; the granules sometimes

fusing together closer to 2.4 or 2.B and then enlarging to ~1 um; annuli

with suspended gemma/columellae and yeast-like granule chains resembling

a shower. [The information above based on crushed spores, with sub-layers

somehow separating and granules disaggregating; we presume that the granules

stay more closely together in nature]. SUB-LAYER 2,B concolorous with 2.4,

but seeming darker when superimposed over sub-laminae; 2-3 um thick,

uniformly covered with hyaline to yellow bacula, 1-1.5 um long, <0.5 um wide;

in unbroken spores, the bacula of this lower sub-layer intermingling with the

yeast-like granule chains from the upper sub-layer, generally taller when just

under the annulus pore.

LAYER 3 tightly adhering to 2.B, rarely separating from it and forming a

separate group (B); <1 um thick, hyaline, membranous.

Innermost wall group comprising layers 4, 5 and 6.

Layer 4 hyaline, ca. lum thick; in crushed spores smooth, slightly wavy or

with a granular surface resembling a “beaded” membrane; commonly tightly

adhering to layer 5, but separating or shrinking to form waves under applied

pressure when the innermost group (endospore) is separated.

Layer 5 hyaline; 1 um thick, expanding to 9-30 um (in PVLG) or to 2-5 um

(in PVLG-Melzer’s reagent), thus revealing its amorphous nature; when

Scutellospora tepuiensis sp. nov. (Venezuela) ... 13

Fic. 1. Scutellospora tepuiensis. a) Whole spore mounted in water. b) Young spore crushed and

mounted in PVLG. c) Crushed spore showing the external wall layer 1 and the two 2) layers.

d) Surface view of layer 2 showing the ring-shaped ornamentation or annuli. e) Left: SEM image

of the underside of layer 2,4, showing gemmae radiating from the pores to form the “showers”.

Right: Underside (inner view) of the layer 2, surface, showing the rounded gemmae radiating from

the annulus pores. f) Crushed spore showing the two sub-layers 4 and B that comprise layer 2,. See

the bacula of 2, B (arrowed) at lower right.

mounted in water, flexible and membranous and difficult to observe in the

endospore wall.

LAYER 6 (innermost) a thin (<1 um thick) flexible membrane that wrinkles

considerably after crushing when mounted in PVLG but not in water or when

intact.

In Melzer’s layers 5 and 6 becoming red-purple while layer 2, turning deep

yellow with the spore content partly yellow; after 24 hours the color reaction

14 ... De Andradeft, Furrazola & Cuenca

Collumellae Gemmae

Ring shaped ornamentation

in surface view

a Component 1

Sub-component 20A

ull neva T

<— Sub-component 20B

en <—— Component 3

Fic. 2. Schematic representation of outermost wall layers of Scutellospora tepuiensis. Annular lips

are defined by either columellae (at left) or gemmae (at right) radiating from a pore.

beginning to fade and after 3-4 days layer 5 turning pink to light purple and

layer 6 remaining red-purple. Other layers do not react.

SPOROPHORE bulbous, 27-43 um diam., 40-57 um tall (from spore base to

the subtending hypha septum); wall ornamented or smooth, contiguous with

spore layers 1 and 2, thickness reaching 4-6 um (<9 um near spore base), with

one stout (10-35 um) projection connected to a slender hypha.

GERMINATION SHIELD (on layer 4) broadly lobed, simple, hyaline, 87-110 x

30-65 um, thin (<1 um thick) walled.

AUXILIARY CELLS: unknown.

DISTRIBUTION & ECOLOGY—Known only from the summit of Sororopan-

tepui, La Gran Sabana, Venezuela. Spores have been collected only from the

low high-tepui shrubland and from the saxicolous community growing on

sandstone at the summit. Soils are acidic and very low in exchangeable P

(0.78-1.8 ppm).

MYCORRHIZAL ASSOCIATIONS unknown. Attempts to form mycorrhizas in

pure culture failed, although the species sporulated abundantly in a multispecies

pot culture with Vigna luteola as host.

ADDITIONAL COLLECTIONS EXAMINED: VENEZUELA, LA GRAN SABANA REGION,

Sororopan-tepui, sclerophyllous shrubland, Dec. 1996, Z. De Andrade, De Andrade-110

Scutellospora tepuiensis sp. nov. (Venezuela) ... 15

Fic. 3. Scutellospora tepuiensis. a) Crushed spore showing germination shield (gs) in lateral view.

b) Crushed spore showing the inner group (4-6) of membranous layers and illustrating the

amorphous nature of layer 5. c) Crushed spore showing the Melzer’s reaction in layers 5 and 6.

d) Bulbous suspensor showing its two layers contiguous with the spore layers. e) Ornamented

bulbous suspensor. The ornamentation is not present in all specimens. f) Partial view of the

germination shield.

(De Andrade personal herbarium); undiluted soil, trap culture (IVIC-38) with Vigna

luteola as host plant, Dec. 1996, Z. De Andrade, De Andrade-118 (De Andrade personal

herbarium); pioneer vegetation, Dec. 1996, De Andrade-123-(1/4) (De Andrade

personal herbarium).

Discussion

The taxonomy and systematics of AMF species forming a bulbous suspensor

and differentiating germinal walls (scutellosporoid spores) have changed

dramatically over recent years. Oehl et al. (2008), who revised Scutellospora

16 ... De Andradet, Furrazola & Cuenca

based on morphological and genetic characters, split the genus into five genera

and three families by weighting germination shield attributes as a diagnostic

character. However, there are overlaps in germination shield morphology

among these segregate genera. Analyses using different molecular tools and

cladistics of 23 morphological characters by Morton & Msiska (2010) did not

support most genera and families of Oehl et al. (2008) but indicated instead

that species producing spores with a bulbous suspensor belong in a single

family, Gigasporaceae, with only three genera: Scutellospora, Gigaspora, and

Racocetra. More recently Redecker et al. (2013), who revised the classification of

arbuscular mycorrhizal fungi—particularly the new taxa proposed by Oehl et al

(2008)—accepted Dentiscutata and provisionally accepted Cetraspora (pending

further study) but rejected Quatunica, Intraornatospora, and Paradentiscutata

and retained all taxa of uncertain affinity in Scutellospora.

For S. tepuiensis, we were unable to see the germination shield in plane

view as required by Oehl et al. (2008) for generic identification, although

what we did see indicates that it is broadly lobed, a trait shared by at least two

genera, Cetraspora and Racocetra. In addition, S. tepuiensis spores comprise

six layers (or components) arranged in two or three groups, as found in most

new genera erected by Oehl et al. (2008). Only Racocetra spores have one inner

group of layers called a germinal wall (Morton & Msiska 2010). In the absence

of molecular support for our new species and in view of its relatively simple

germination shield, we prefer to adopt a conservative approach by describing

this species in Scutellospora.

Scutellospora tepuiensis can be distinguished from other scutellosporoid

species by its extremely complex ornamentation. Only Dentiscutata reticulata

(Koske et al.) Sieverd. et al. has an ornamentation of such complexity. In addition,

mature spores of both species are similar in color. However, S. tepuiensis spores

have an annulate ornamentation, while D. reticulata spores are covered with

a reticulum overlaying spines. Dentiscutata nigra has rounded holes in the

outer wall, but its spores are much larger (~500 um) and darker-colored (dark

brown to black) than those of S. tepuiensis. Other scutellosporoid species with

a remarkable ornamentation are S. crenulata R.A. Herrera et al. (with spores

covered with dome-like papillae) and S. striata Cuenca & R.A. Herrera (with

spore ornamentation resembling a fingerprint). The other ornamented species

retained in Scutellospora after the papers by Morton and Msiska (2010) and

Redecker et al. (2013) are S. nodosa Blaszk. (with the outermost layer forming a

knobbed spore surface) and S. spinosissima (with the laminate spore wall layer

2 ornamented with spines).

Three inner layers similar to those of S. tepuiensis have also been noted

in other Scutellospora species, e.g., S. spinosissima (Walker et al. 1998),

Scutellospora tepuiensis sp. nov. (Venezuela) ... 17

S. projecturata Kramad. & C. Walker (Kramadibrata et al. 2000), S. striata

(Cuenca & Herrera-Peraza 2008), and S. crenulata (Herrera-Peraza et al.

2001). According to Walker et al. (1998), the morphological features of the

innermost wall layers of this fungal group probably differ from germinal wall 2

of other scutellosporoid species. In addition the layers forming this inner group

frequently tightly adhere to each other and thereby this wall may resemble a

structure consisting of one coriaceous layer (Walker et al. 1998).

Layer 4 of the inner wall group B or C of S. tepuiensis spores occasionally

resembles a beaded layer commonly occurring in the innermost spore wall

in species of Acaulosporaceae. A similar layer was also identified in spores of

S. crenulata described from specimens found in La Gran Sabana (Herrera-

Peraza et al. 2001). Molecular and cladistic morphological analyses support

Gigasporales as more closely related to Acaulosporaceae than to Glomerales

(Morton & Benny 1990). The presence of a beaded layer in S. crenulata and

S. tepuiensis additionally supports this finding.

Future sampling of S. tepuiensis at the summit of Venezuelan tepuis will

enable us to conduct the necessary molecular analysis that should indicate

its phylogenetic position within Scutellospora. Nevertheless, its unique

ornamentation when compared to all other described Scutellospora species

distinguishes S. tepuiensis within Gigasporaceae. The La Gran Sabana ecosystem

has become an important one for discovering new AMF species.

Acknowledgements

We are in debt to our late fellow Zita De Andrade, who discovered S. tepuiensis during

a short trip to the Sororopan-tepui, and to late Dr. Ricardo Herrera-Peraza and Roberto

Ferrer from the Institute of Ecology and Systematics of La Habana, Cuba, who prepared

the first draft of species description. Milagros Lovera gave invaluable comments to the

manuscript. We thank Drs. J. Blaszkowski and S. Stiirmer for reviewing the manuscript.

We are also grateful to Mycotaxon, Ltd. for underwriting the cost of this publication.

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82: 721-725. http://dx.doi.org/10.1006/anbo.1998.0728

MY COTAXON

January-March 2017— Volume 132, pp. 19-27

http://dx.doi.org/10.5248/132.19

Five penzigioid Xylaria species from Veracruz (Mexico)

FrIpe Tapia’, Yu-MING Ju’,

SANTIAGO CHACON’ & ELSA UTRERA-BARILLAS'

"Instituto de Ecologia, A.C. Apartado Postal 63, Xalapa, Veracruz 91000, México

*Institute of Plant and Microbial Biology, Academia Sinica, Nankang, Taipei 115, Taiwan

*CORRESPONDENCE TO: santiago.chacon@inecol.mx

ABSTRACT—Five species of the genus Xylaria with penzigioid stromata were collected from

the state of Veracruz, Mexico. Xylaria albocinctoides, X. discolor, and X. xylarioides are new

records for the Mexican mycobiota; X. frustulosa is a new record for Veracruz State; and

X. berteroi is recorded from new localities within the state. Photographs of the stromata of

the five species are presented, as well as a dichotomous key to the penzigioid Xylaria species

known in Mexico.

KEY worDs—Ascomycota, taxonomy, chorology

Introduction

Xylaria Hill ex Schrank is the type genus of the family Xylariaceae, with

approximately 300 species (Kirk et al. 2008), some of which are characterized

by penzigioid stromata. Penzigioid Xylaria were once placed in Penzigia Sacc.,

but Ju & Rogers (2001) showed that the type species, P cranioides Sacc. &

Paol., belongs to Xylaria. There are more than 40 penzigioid Xylaria species

worldwide (Ju et al. 2012), characterized by sessile stromata and adherence to

the substrate by a narrow central connective.

San Martin & Rogers (1995) recorded 105 Xylaria species in Mexico. Of

the 109 species listed by Medel (2007), 45 are from Veracruz State (Medel et

al. 2008). Penzigioid Xylaria species previously known from Mexico include

X. anisopleura (Mont.) Fr., X. berteroi, X. boergesenii (Ferd. & Winge)

P.E Cannon, X. enteroleuca (J.H. Mill.) PM.D. Martin, and X. frustulosa

(San Martin & Rogers 1995). Chacon & Guzman (1983) recorded “Penzigia

20 ... Tapia & al.

conostoma” (a name they attributed to J.H. Miller) from Veracruz, but Miller's

(1961) “Hypoxylon conostomum” is a misdetermination of X. boergesenii (fide

Cannon 1987).

The present study contributes to a better understanding of Xylaria species

with penzigioid stromata from Veracruz State.

Materials & methods

We examined approximately 50 specimens of Xylaria with penzigioid stromata

(all from localities in central Veracruz State), of which most were collected by the authors

and a few are collections made by others and deposited at the Herbarium, Instituto de

Ecologia, A.C., Xalapa, Mexico (XAL). The stromata were sectioned and mounted in

5% KOH or Melzer’s reagent. Descriptions are given for X. albocinctoides, X. discolor,

and X. xylarioides (newly recorded from Mexico) and for X. frustulosa, not previously

described based on Mexican material. Photographs of stromata are presented. All of

the studied specimens were dried, labelled, and deposited at XAL, and duplicates were

deposited at the Herbarium, Biodiversity Research Center, Academia Sinica, Taipei,

Taiwan (HAST).

Taxonomy

Xylaria albocinctoides Y.M. Ju, H.M. Hsieh & J.D. Rogers,

Mycologia 104: 768 (2012) PLATE 1A

STROMATA superficial, gregarious, aggregated to dispersed, sometimes

confluent, semicircular, flattened to pulvinate, occasionally subglobose,

reddish brown to blackish brown, slightly shiny, 0.5-2 mm diam x 0.5-0.8

mm thick, <10 mm long when confluent, attached to substrate by a narrow

central connective, with 4-20 perithecia enclosed within each stroma, with

white interior. PERITHECIA globose to subglobose, 0.2-0.3 mm diam. OSTIOLES

black, papillate, 0.1 mm broad, slightly raised. Asci1 with eight ascospores

arranged in uniseriate manner, hyaline, cylindrical, 150-170 x 12-16 um, with

an amyloid apical ring. Ascosporss (18-)19-23 x 10-12 um, dark brown to

blackish brown, ellipsoid, inequilateral, somewhat rounded at ends, with a

straight germ slit spore-length.

Hasitat—On dead branches of Piper sp. and secondary vegetation in a

montane cloud forest at altitudes between 1300 and 1550 m.

SPECIMENS EXAMINED—MEXICO, VERACRUZ STATE, Municipality of Xalapa, Cerro

Macuiltepec Ecological Park, 9 August 2011, S. Chacon, E. Utrera & F. Tapia 6204; 6

September 2011, S. Chacon & F. Tapia 6349; 27 September 2011, S. Chacon & F. Tapia

6387, 6388; 11 October 2011, S$. Chacén & F. Tapia 6461; 25 October 2011, S. Chacon

& E. Tapia 6533; 6 March 2013, S. Chacon & F. Tapia 6945; 10 April 2013, S. Chacon

7055-B; 11 July 2013, S$. Chacén & FE. Tapia 7207; 30 July 2013, S. Chacon & F. Tapia

7290; 5 August 2014, Chacon & F. Tapia 7699; INECOL Cloud Forest Sanctuary, Km 2.5

Penzigioid Xylaria spp. new for Mexico... 21

old road Xalapa—Coatepec, 25 August 2010, S. Chacon & E. Utrera 5929; 9 September,

2010, S. Chacon, E. Utrera & M. Soto 5970; 24 November 2010, S. Chacon & E. Utrera

6176; 22 August 2011, S. Chacon & E. Utrera 6246; 26 August 2011, S. Chacon, F. Tapia

& E. Utrera 6292; 14 October 2011, S. Chacon 6477; 21 October 2011, S. Chacon & E

Tapia 6522; 28 October 2011, S. Chacon & F. Tapia 6546; 3 April 2014, F. Tapia 3008; 17

May 2012, S. Chacon & FE. Tapia 6790-B; Tecajetes Park, 3 July 2013, S. Chacon, F. Tapia

& E. Utrera 7154; Francisco Javier Clavijero Ecological Park, Km 2.5 old road Xalapa-

Coatepec, 8 December 2003, S. Chacon 5506.

REMARKS—Xylaria albocinctoides is a penzigioid species with a thin, soft

stromatal surface. Macroscopically it is similar to X. albocincta (Rehm)

Y.M. Ju et al., which differs mainly by larger ascospores 27-32 x 12-14 um.

The Mexican X. albocinctoides specimens have ascospore measurements very

similar to those given in Ju et al. [2012; (18-)19-21(-22.5) x 10-11.5 um].

Xylaria albocinctoides, previously known only from Venezuela, represents

a new record for Mexico and is the most frequently encountered penzigioid

Xylaria species recorded from Veracruz.

Xylaria berteroi (Mont.) Cooke ex J.D. Rogers & Y.M. Ju,

N. Amer. Fung. 7(9): 18 (2012) PLATE 1B

= Penzigia enteroleuca J.H. Mill., Mycologia 32: 183 (1940)

Hasitat—On dead wood of angiosperms in a montane cloud forest, in a

coffee farm within a secondary vegetation at altitudes of 1300 to 1600 m.

SPECIMENS EXAMINED—MEXICO, VeERACRUZ STATE, Municipality of Xalapa,

San Roque Hills, 8 August 2012, F. Tapia & S. Chacén 2615; INECOL Cloud Forest

Sanctuary, Km 2.5 old road Xalapa—Coatepec, 3 September 2010, S. Chacon & E. Utrera

5948; 1 October 2010, S. Chacén & E. Utrera 6049; 14 October 2011, S. Chacon & F.

Tapia 6473; 6 June 2013, S. Chacon 7103; 19 June 2013, S. Chacon & F. Tapia 7109, 7128;

2 July 2013, S. Chacon & E. Tapia 7147, 8 August 2013, S. Chacon, E. Utrera & F. Tapia

7342, 7349; Natura Park protected area, 8 October 2011, S. Chacon & F. Tapia 6502; 27

February 2013, S. Chacon 6930; 3 April 2013, S. Chacén 7008, 7009, 7012, 7015, 7021;

19 July 2013, S. Chacon & F. Tapia 7233; Francisco Javier Clavijero Ecological Park,

Km 2.5 old road Xalapa—Coatepec, 12 May 1982, Brown 416; 8 June 1983, S. Chacon

1068; 12 December 1983, S. Chacon 1934; 7 May 1986, L. Montoya 530; 23 March 1987,

R. Medel 329; Francisco Javier Clavijero Botanical Garden, Km 2.5 old road Xalapa-

Coatepec, 17 July 1991, S. Chacén 4519; 9 August 1995, S. Chacon 4925; 14 June 2000, R.

Medel 756; 11 September 2006, R. Medel 1315; 2 September 2011, S. Chacon & E. Utrera

6331; 2 July 2012, FE Tapia 2576; 24 October 2012, F. Tapia 2691; Around the child care

house CONECALLI, Km 2.5 old road Xalapa—Coatepec, 3 December 1990, FE. Tapia 431;

Cerro Macuiltepec Ecological Park, 6 September 2011, S. Chacon 6349-A; 27 September

2011, S. Chacon & FE. Tapia 6390; 6 March 2013, S. Chacén 6935, 6948; 10 April 2013, S.

Chacon 7065; 11 July 2013, S$. Chacon & F. Tapia 7204, 7208; 30 July 2013, S. Chacon &

E Tapia 7279, 7285. Municipality of San Andrés Tlalnehuayocan, Agiiita Fria Ranch,

6 April 2007, R. Medel 1351. Municipality of Acatlan, top Acatlan Volcano, 2 October

2013, S. Chacon, F. Tapia & E. Utrera 7419. Municipality of Coatepec, about 1 km of the

road Coatepec-El Grande, 23 October 2012, F. Tapia 2686-B.

22 ... Tapia & al.

REMARKS—Xylaria berteroi can be diagnosed by its more or less pulvinate to

discoid stromata (0.7—2.5 cm diam x 0.2-0.7 cm thick), a thick hard carbonaceous

outer stromatal layer (sometimes cracked on the surface), and ascospores

[(9-)12-13.5(-14) x (6-)6.5-7.5(-8) um] with a straight germ slit running

almost the entire ascospore length. Welden et al. (1979) and Medel & Chacon

(1997) reported X. berteroi from Veracruz State, and our research extends

its known distribution to other localities in central Veracruz. The specimen

“Brown 416”, cited by Chacon & Guzman (1983) as Penzigia enteroleuca,

corresponds to X. berteroi; these two species are difficult to separate—

Callan & Rogers (1990) suggested that they are probably conspecific, and

Rogers & Ju (2012) treated them as conspecific.

Xylaria discolor (Berk. & Broome) Y.M. Ju, H.M. Hsieh, J.D. Rogers & Jaklitsch,

Mycologia 104: 772 (2012) PLATE 1C,D

= Penzigia bermudensis J.H. Mill., Mycologia 32: 402 (1940)

STROMATA erumpent to superficial, dispersed, gregarious to confluent,

subglobose to pulvinate, sometimes flattened, whitish grey to dark grey due to

the presence of a outer layer, which falls off with age, leaving the surface a dark

brown to blackish brown, 0.3-2 mm diam. x 0.5 mm thick, <1.5 cm long when

confluent, attached to the substrate by a narrow central connective, containing

<25 perithecia per stroma; surface dotted with ostioles; interior soft, yellow,

whitish towards the base. PERITHECIA globose to subglobose, 0.1-0.3 mm

diam. OsTIo Es black, papillate, 0.1 mm broad, slightly raised. Asci with eight

uniseriately arranged ascospores, hyaline, cylindrical, 90-140 x 8-10 um, with

an inamyloid apical ring. Ascosporgs dark brown to blackish brown, ellipsoid,

equilateral, with rounded apices, (8-)9-12 x 5.5-7.5(-8) um, with a straight,

spore-length germ slit.

Hasitat—On dead wood in a montane cloud forest and in secondary

vegetation of an abandoned coffee plantation. Altitudes 1300 to 1450 m.

SPECIMENS EXAMINED—MEXICO, VERACRUZ STATE, Municipality of Xalapa,

Francisco Javier Clavijero Botanical Garden, Km 2.5 old road Xalapa—Coatepec, 7

October 2010, S. Chacon & E. Utrera 6076; Palo Verde pantheon of Xalapa, 21 March

2012, S. Chacén 6984; Cologne El Olmo, former hacienda Las Animas, 17 April 2013,

S. Chacon 7092; Cerro Macuiltepec Ecological Park, 28 August 2014, S. Chacon & F.

Tapia 7783.

REMARKS—Macroscopically X. discolor is somewhat similar to X. lechatii

Y.M. Ju et al., but the latter species has a white interior, an apical ring staining

pale blue only at base in Melzer’s iodine reagent and larger ascospores 12-15

x 6.5-8 um. According to Ju et al. (2012) and Rogers & Ju (2012), Penzigia

bermudensis is a synonym of X. discolor. Seaver & Waterston (1940) described

P. bermudensis with ascospores of 8-10 x 6-8 um; our measurement of the

Penzigioid Xylaria spp. new for Mexico ... 23

PLaTE 1. Stromata of Xylaria species. X. albocinctoides: A. scale bar = 2 mm. X. berteroi: B. scale

bar = 7 mm. X. discolor: C. scale bar = 1.3 mm, D. scale bar = 1 mm. X. frustulosa: E. scale bar =

1.5 mm. X. xylarioides: F. scale bar = 1 mm, G. scale bar = 0.7 mm.

Mexican X. discolor ascospores agrees with those in Ju et al. (2012); (8.5-)9-12

x 5.5-7(-7.5) um and Rogers & Ju (2012; 9-10.5 x 6-8 x 5-6 um). This is the

first record of X. discolor from Mexico.

Xylaria frustulosa Y.M. Ju, H.M. Hsieh & J.D. Rogers, (Berk. & M.A. Curtis) Cooke,

Grevillea 12: 5 (1883) PLATE 1E

= Sarawakus frustulosus (Berk. & M.A. Curtis) Lar. N. Vasiljeva, Nizshie

Rasteniya, Griby i Mokhoobraznye Dalnego Vostoka Rossii 4: 157 (1998)

24 ... Tapia & al.

STROMATA superficial, gregarious to confluent, semicircular to irregular,

flattened to pulvinate, dark brown to black-brown, 0.5-2.7 mm diam. x 0.5-0.8

mm thick, <8 mm long when confluent, attached to the substrate by a narrow

central connective, containing numerous perithecia; surface dotted with

ostioles; interior soft, white. PERITHECIA globose to subglobose, 0.1-0.2 mm

diam. OstIoxzs black, papillate, 0.1 mm wide, slightly raised. Asci with eight

ascospores, hyaline, cylindrical, 40-50 x 4-6 um, with a long stipe, and a small

amyloid apical ring. Ascosporss pale greenish brown, elliptical, 5-6(-7) x

2-3 um, with an inconspicuous germ slit.

HasBitat—On decorticated wood in a Mangifera indica plantation, alt.

500 m.

SPECIMEN EXAMINED—MEXICO, VERACRUZ STATE, Municipality of Jalcomulco,

beside restaurant El Rey, road Tlaltetela-~Xalapa, 6 October 2012, S. Chacén 6869-B.

REMARKS—Xylaria frustulosa is characterized mainly by its subdiscoid

confluent stromata with small (no more than 0.8 mm) diameters and small

ascospores with barely discernible germ slits. The ascospore size range of the

Mexican material in general agrees that given by Miller (1934), Dennis (1970),

Martin (1970), Jong & Rogers (1970), and Ju et al. (2012). The combination of

this species in Sarawakus Lloyd (Vasilyeva 1998) is untenable; Sarawakus has a

hypocrealean type species and is considered to be a synonym of Trichoderma

(Samuels & Rossman 1992, Jaklitsch et al. 2014).

Xylaria frustulosa is known from tropical and subtropical regions in the

Americas and Asia. San Martin & Rogers (1996) cited X. frustulosa (without a

description) from Tamaulipas State, Mexico. Our collection represents the first

record of X. frustulosa from Veracruz State.

Xylaria xylarioides (Speg.) Hladki & A.I. Romero,

Fungal Diversity 42: 86 (2010) PLATE 15,G

STROMATA Solitary to gregarious, the fertile part conical to subrhomboid or

subcylindrical, 0.8-3 x 0.6-2 mm, with perithecial contours well pronounced

and with the apex conical and sterile, sessile or sometimes with a short stipe <1.5

mm long, tomentose, concolorous with the fertile part; peeling layer arranged

in pale brown bands, running from apex to middle. PERITHECIA globose to

subglobose, 0.2-0.7 mm diam. OsTIoLEs shiny black, papillate, small, up to

0.1 mm diam; surface roughened, black to blackish brown; interior soft, white.

AscI containing eight uniseriately arranged ascospores, hyaline, cylindrical,

140-195 x 8-9 um, with an amyloid apical ring. Ascospores brown to blackish

brown, ellipsoid to navicular, inequilateral, with apices more or less rounded

or mucronate, (17—)19-22 x (6.5-)7-9.5 um, with a straight germ slit running

almost the entire spore length.

Penzigioid Xylaria spp. new for Mexico ... 25

Hasitat—On dead branches of unidentified angiosperms, in a montane

cloud forest and natural secondary vegetation at altitudes of 1300 to 1600 m.

SPECIMENS EXAMINED—MEXICO, VERACRUZ STATE, Municipality of Xalapa, Cerro

Macuiltepec Ecological Park, 11 October 2011, S. Chacén & F. Tapia 6459; 11 July 2013,

S. Chacén 7199; INECOL Cloud Forest Sanctuary, Km 2.5 old road Xalapa—Coatepec,

30 September 2011, S. Chacon & E. Utrera 6419; S. Chacon & F. Tapia 6420; Tecajetes

Park, 1 August 2013, S. Chacon, E. Utrera & F. Tapia 7305; Francisco Javier Clavijero

Ecological Park, Km 2.5 old road Xalapa—Coatepec, 24 April 2014, F. Tapia & S. Chacon

3023.

REMARKS— his species is characterized by the presence of a conical sterile

apex, a well-developed peeling layer, and ascospore size and shape. The Mexican

material agrees with the description of Hladki & Romero (2010; ascospores

17-21 x 6.5-9 um). The Mexican material of X. xylarioides shows a great

variability in stipe length; in some collections the stipe is almost indiscernible

but in others it reaches up to 3 mm long.

Dichotomous key to penzigioid Xylaria species in Mexico

1. Apex of the stromata conical, sterile;

ascospores. 17-22 * 6,5-9:5 Ui. i540 cnee eden sy ecnyt end eennyon X. xylarioides

lanexcof stromata flattened-oriconvex, erties wwa.8 fees fecet feacek het tet oe 2.

ZU ASCOSPOLES 22 OS UNT OMG 5 sy on obs gn a¥ ein 6 «af eicnie nabs ule mses eer nibs eghcaln te ogee al pete. oe i!

2. ASCOSPORES 20M LONG Aat ao. Aa a oy tenn sk een doen ares Naess Matera s Nees +

BeStromata flattenechto pulhanate sd... Wi aad oe er ead en kak ier Bod otetad enamel. whanad cd 6

3. Stromata subglobose to elongate, 0.4-4 cm high;

ASCOSPOLES ZO SIO: 229 We S4HINN fa. op Fic dhenscapther ena at sae skh ost a othe X. anisopleura

4, Stromata 1-2 cm diam, with a carbonaceous outer layer;

TSG BAS 0201 eka: Eel Re Rea Oar 0 WU 18 Ae Mh ns RA nA eR A VAAL Ri ALT Sty TER SENSE X. berteroi

4, Stromata 0.5 mm diam, with a thin and soft outer layer .................-.-000- 5

5. Stromatal interior yellow; ascospores 8-12 x 5.5-8 uM .............04. X. discolor

5. Stromatal interior white; ascospores 5-7 x 2-3 um _ .............--5- X. frustulosa

6. Ascospores 18-23 x 10-12 um, with a straight germ slit .......... X. albocinctoides

6. Ascospores 19.5-26 x 7-8.5 «um, with a spiral to sigmoid germ slit ... X. boergesenii

Acknowledgements

We thank the authorities of the Instituto de Ecologia, A.C. of Xalapa, Veracruz, for

financial support: Patterns of Species Diversity in Urban Zones (grant 20035 30842) and

Integral Study 2013-2037 on biodiversity of Francisco Javier Clavijero Botanical Garden,

with emphasis on the cloud forest sanctuary (grant 20035-30890). The authors would

like to express their gratitude to Dr. Andrea I. Romero (Universidad de Buenos Aires,

Argentina) and Dr. Fidel Landeros (Universidad Aut6noma de Querétaro, México) for

reviewing the manuscript and for helpful comments. We thank Manuel Hernandez

26 ... Tapia & al.

for preparing the figure plate and Juan Lara Carmona for his help in curatorial and

technical activities.

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MY COTAXON

January-March 2017— Volume 132, pp. 29-42

http://dx.doi.org/10.5248/132.29

Three species of wood-decaying fungi in Polyporales

new to China

CHANG-LIN ZHAO‘, SHI-LIANG LIu*, GUANG-JUAN REN,

XIAO-HONG JI & SHUANGHUI HE*

Institute of Microbiology, Beijing Forestry University,

No. 35 Qinghuadong Road, Haidian District, Beijing 100083, PR. China

* CORRESPONDENCE TO: shuanghuihe@yahoo.com

ABSTRACT— Three wood-decaying fungi, Ceriporiopsis lagerheimii, Sebipora aquosa, and

Tyromyces xuchilensis, are newly recorded in China. The identifications were based on

morphological and molecular evidence. The phylogenetic tree inferred from ITS+nLSU

sequences of 49 species of Polyporales nests C. lagerheimii within the phlebioid clade,

S. aquosa within the gelatoporia clade, and T: xuchilensis within the residual polyporoid clade.

The three species are described and illustrated based on Chinese material.

Key worps—Basidiomycota, polypore, taxonomy, white rot fungus

Introduction

Wood-decaying fungi play a key role in recycling nutrients of forest

ecosystems by decomposing cellulose, hemicellulose, and lignin of the plant

cell walls (Floudas et al. 2015). Polyporales, a large order in Basidiomycota,

includes many important genera of wood-decaying fungi. Recent molecular

studies employing multi-gene datasets have helped to provide a phylogenetic

overview of Polyporales, in which thirty-four valid families are now recognized

(Binder et al. 2013).

The diversity of wood-decaying fungi is very high in China because of the

large landscape ranging from boreal to tropical zones. More than 1200 species

of wood-decaying fungi have been found in China (Dai 2011, 2012), and some

* Chang-lin Zhao and Shi-liang Liu contributed equally to this work

and share first-author status

30 ... Zhao, Liu & al.

important genera have been extensively investigated (Jia & Cui 2011; He & Dai

2012; Zhou & Dai 2012, 2013; Li & Cui 2013; Li et al. 2014; Song et al. 2014;

Chen et al. 2015, 2016; Zhou 2015; Zhou et al. 2015, 2016a,b,c). Recently, three

additional species in Polyporales—Ceriporiopsis lagerheimii, Sebipora aquosa,

and Tyromyces xuchilensis—were found in Yunnan Province, southwestern

China, and are described and illustrated here. In addition to morphological

analysis, the phylogenetic positions of these Chinese specimens were inferred

from ITS+nLSU sequences.

Materials & methods

Morphological study

The studied specimens are deposited at the herbaria of Beijing Forestry University,

Beijing, China (BJFC) and University of Oslo, Oslo, Norway (O). Macro-morphological

descriptions are based on field notes. Special colour terms follow Petersen (1996).

Micro-morphological data were obtained from the dried specimens under a light

microscope. The following abbreviations are used: KOH = 5% potassium hydroxide,

CB = Cotton Blue, CB- = acyanophilous, IKI = Melzer’s reagent, IKI- = neither amyloid

nor dextrinoid, L = mean spore length, W = mean spore width, Q = variation in the

L/W ratios, n = number of spores (a) measured from number of specimens (b).

Molecular phylogeny

A CTAB rapid plant genome extraction kit-DN14 (Aidlab Biotechnologies Co.,

Ltd, Beijing) was used to obtain PCR products from dried specimens, according to the

manufacturer's instructions with some modifications. ITS region was amplified with

the primer pair ITS5 and ITS4 (White et al. 1990), and the nuclear LSU region was

amplified with the primer pair LROR and LR7 (http://www.biology.duke.edu/fungi/

mycolab/primers.htm). The ITS PCR began with initial denaturation at 95°C for 3 min,

followed by 35 cycles at 94°C for 40 s, 58°C for 45 s, and 72°C for 1 min, and a final

extension at 72°C for 10 min. The nLSU PCR began with initial denaturation at 94°C for

1 min, followed by 35 cycles at 94°C for 30 s, 48°C for 1 min, and 72°C for 1.5 min, and

a final extension at 72°C for 10 min. The PCR products were purified and sequenced at

Beijing Genomics Institute. The sequences were deposited at GenBank (TABLE 1).

Sequencher 4.6 (GeneCodes, Ann Arbor) was used to edit the DNA sequences.

The original ITS and nLSU sequences were combined directly and then aligned in

MAFFT 6 (Katoh & Toh 2008, http://mafft.cbrc.jp/alignment/server/) using the

“G-INS-I” strategy and manually adjusted in BioEdit (Hall 1999). The concatenated

alignment was subjected to the incongruence length difference (ILD) test (Farris et al.

1994) implemented in PAUP* 4.0b10 (Swofford 2002) with a heuristic search and 1000

bootstrap (BS) replicates. The ILD test generated a P value of 1.000 much greater than

0.01, indicating that there was no incongruence between the ITS and nLSU regions. The

sequence alignment was deposited in TreeBase (submission ID 19179). Heterobasidion

annosum (Fr.) Bref. and Stereum hirsutum (Willd.) Pers. were selected as outgroup

(Binder et al. 2013). Clade names follow Binder et al. (2013).

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 31

TABLE 1. Species and sequences used in the phylogenetic analyses.

Newly generated sequences are set in bold.

SPECIES

Abortiporus biennis (Bull.) Singer

Antrodia albida (Fr.) Donk

Antrodia heteromorpha (Fr.) Donk

Antrodiella americana Ryvarden & Gilb.

Antrodiella semisupina (Berk. & M.A. Curtis)

Ryvarden

Ceriporiopsis balaenae Niemela

Ceriporiopsis consobrina (Bres.) Ryvarden

Ceriporiopsis fimbriata C.L. Zhao & Y.C. Dai

Ceriporiopsis gilvescens

Ceriporiopsis guidella

Ceriporiopsis lagerheimii

Climacocystis borealis

Coriolopsis caperata (Berk.) Murrill

Dacryobolus karstenii (Bres.) Oberw. ex Parmasto

Daedalea quercina (L.) Pers.

Earliella scabrosa (Pers.) Gilb. & Ryvarden

Fomitopsis pinicola (Sw.) P. Karst.

Fomitopsis rosea (Alb. & Schwein.) P. Karst.

Fragiliporia fragilis Y.C. Dai et al.

Ganoderma lingzhi Sheng H. Wu et al.

Gelatoporia subvermispora

Grammothelopsis subtropica B.K. Cui & C.L. ZhaO

Heterobasidion annosum

Hornodermoporus martius (Berk.) Teixeira

VOUCHER NO.

TERI 274

CBS 308.82

CBS 200.91

Gothenburg 3161

FCUG 960

H7002389

Rivoire 977

Dai 11672

Cui 1671

BRNM 667882

BRNM 710166

Yuan 2752

HUBO 7659

Ryvarden 58240

Dai 12304

KH 13318

LE(BIN)-0677

KHL 11162

DSM 4953

PR1209

CBS 221.39

ATCC 76767

Dai 13080

Dai 13559

Dai 13561

Wu 1006-38

BRNU 592909

Cui 9041

PFC 5252

MUCL 41677

GENBANK NO.

ITS

EU232187

DQ491414

DQ491415

JN710509

EU232182

FJ496669

FJ496667

KJ698633

KJ698634

FJ496685

FJ496684

KF845946

FJ496687

KX081077

KX161647

JQ031126

AB158316

EU118624

DQ491425

JN165009

DQ491405

DQ491410

KJ734260

KJ734261

KJ734262

JQ781858

FJ496694

JQ845096

KC492906

FJ411092

nLSU

EU232235

AY515348

AY515350

JN710509

EU232266

FJ496717

FJ496716

KJ698637

KJ698638

FJ496719

FJ496720

KF845953

FJ496722

KX161652

KX161651

JQ031126

AB158316

EU118624

DQ491425

JN164793

DQ491405

DQ491410

KJ734264

KJ734265

KJ734266

FJ496706

JQ845099

KC492906

FJ393859

32 ... Zhao, Liu & al.

TABLE 1, concluded

SPECIES

Hypochnicium lyndoniae (D.A. Reid) Hjortstam

Junghuhnia nitida (Pers.) Ryvarden

Obba rivulosa (Berk. & M.A. Curtis)

Miettinen & Rajchenb.

Obba valdiviana (Rajchenb.) Miettinen & Rajchenb.

Perenniporia medulla-panis (Jacq.) Donk

Perenniporiella neofulva (Lloyd) Decock & Ryvarden

Phlebia livida (Pers.) Bres.

Phlebia radiata Fr.

Phlebia subserialis (Bourdot & Galzin) Donk

Piloporia sajanensis (Parmasto) Niemela

Podoscypha venustula (Speg.) D.A. Reid

Polyporus tuberaster (Jacq. ex Pers.) Fr.

Postia guttulata (Sacc.) Jiilich

Sebipora aquosa

Skeletocutis amorpha (Fr.) Kotl. & Pouzar

Skeletocutis jelicii Torti¢ & A. David

Skeletocutis portcrosensis A. David

Skeletocutis subsphaerospora A. David

Steccherinum fimbriatum (Pers.) J. Erikss.

Steccherinum ochraceum (Pers.) Gray

Stereum hirsutum

Truncospora ochroleuca (Berk.) Pilat

Tyromyces chioneus (Fr.) P. Karst.

Tyromyces xuchilensiS

Xanthoporus syringae (Parmasto) Audet

VOUCHER NO.

NL 041031

KHL 11903

KCTC 6892

FF 503

MUCL 49581

MUCL 45091

FCUG 2189

UBCF 19726

FCUG 1434

Mannine 2733a

CBS 65684

CulTENN 8976

KHL 11739

Miettinen 8868

Miettinen 8680

Miettinen 9265

Dai 13268

Dai 13592

Miettinen 11038

H 6002113

LY 3493

Rivoire 1048

KHL 11905

KHL 11902

NBRC 6520

MUCL 39726

Cui 10225

Dai 12234

Ryvarden 44669

Gothenburg 1488

GENBANK NO.

ITS

JX124704

EU118638

FJ496693

HQ659235

FJ411088

FJ411080

AF141624

HQ604797

AF141631

HQ659239

JN649367

AF516598

EU11865

HQ659242

HQ659240

HQ659243

KX161648

KU376422

FN907913

F}496690

FJ496689

FJ496688

EU118668

JQ031130

AB733150

FJ411098

KF698745

KX161649

KX161650

JN710607

nLSU

JX124704

EU118638

FJ496710

HQ659235

FJ393876

FJ393852

AF141624

HQ604797

AF141631

HQ659239

JN649367

AJ488116

EU11865

HQ659240

KX161661

KX161660

FN907913

FJ496727

FJ496689

FJ496688

EU118668

JQ031130

AB733325

FJ393865

KF698756

KX161658

KX161659

JN710607

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 33

Maximum parsimony analysis was applied to the ITS+nLSU sequences dataset,

and performed in PAUP* version 4.0b10 (Swofford 2002) according to Zhao et al.

(2013). All characters were equally weighted, and gaps were treated as missing data.

Trees were inferred using the heuristic search option with TBR branch swapping and

1000 random sequence additions. Max-trees were set to 5000, branches of zero length

were collapsed, and all parsimonious trees were saved. Clade robustness was assessed

using a bootstrap (BT) analysis with 1000 replicates (Felsenstein 1985). Descriptive

tree statistics, tree length (TL), consistency index (CI), retention index (RI), rescaled

consistency index (RC), and homoplasy index (HI) were calculated for each generated

Maximum Parsimonious Tree (MPT). Sequences were also analyzed using Maximum

likelihood (ML) with RAxML-HPC2 on Abe through the Cipres Science Gateway

(www.phylo.org). Branch support for ML analysis was determined by 1000 bootstrap

replicate.

MrModeltest 2.3 (Posada & Crandall 1998, Nylander 2004) was used to determine

the best-fit evolution model for Bayesian inference (BI). BI was calculated with

MrBayes3.1.2 with a general time reversible (GTR) model of DNA substitution and a

gamma distribution rate variation across sites (Ronquist & Huelsenbeck 2003). Four

Markov chains were run for 10 million generations, and trees were sampled every 100

generations. The first quarter generations were discarded as burn-in. A majority rule

consensus tree of all remaining trees was calculated.

Results

Phylogenetic analyses

The combined dataset (ITS+nLSU) included sequences from 62 samples

representing 51 species. The dataset had an aligned length of 2202 characters,

of which 1294 characters are constant, 232 are variable and parsimony-

uninformative, and 676 are parsimony-informative. MP analysis yielded 13

equally parsimonious trees (TL = 4893, CI = 0.313, HI = 0.687, RI = 0.578,

RC = 0.181). Best model estimated and applied in the Bayesian analysis

was: GTR+I+G, lset nst = 6, rates = invgamma; prset statefreqpr = dirichlet

(1,1,1,1). The average standard deviation of split frequencies in BI was

0.001416. BI and ML analyses resulted in similar topologies as that of MP

analysis.

The MP phylogenetic tree supported seven major clades for 49 species of

Polyporales with Ceriporiopsis lagerheimii nested within the phlebioid clade,

Sebipora aquosa within the gelatoporia clade, and Tyromyces xuchilensis

within the residual polyporoid clade (Fic. 1). For all these three species,

specimens from China clustered with authentic specimens from elsewhere

with strong support (Fie. 1).

34 ... Zhao, Liu & al.

100/100/1.00 ~- Obba valdiviana FF503

Stereum hirsutum NBRC 6520

Heterobasidion annosum PFC 5252 Outgr oup

Fic. 1. Maximum parsimony strict consensus tree illustrating the phylogeny of Ceriporiopsis

lagerheimii, Sebipora aquosa, Tyromyces xuchilensis, and related species in Polyporales based on

ITS+nLSU sequences. Branches are labeled with maximum likelihood bootstrap =70%, parsimony

bootstrap proportions 250% and Bayesian posterior probabilities 20.95.

Taxonomy

Ceriporiopsis lagerheimii Lessoe & Ryvarden, Syn. Fung. 27: 44, 2010. Fies 2a, 3

BASIDIOME Annual, resupinate, adnate, soft corky, without odor or taste

when fresh, corky when drying; <2 cm long, 1.5 cm wide, 2 mm thick at center.

Pore surface cream when fresh, cream to buff upon when drying; pores angular,

5-6 per mm; dissepiments thin, entire. Subiculum thin, white, <0.2 mm thick.

Tubes concolorous with pore surface, <1.8 mm long.

HyPHAL STRUCTURE Monomitic; generative hyphae with clamp connections;

IKI-, CB-; tissues unchanged in KOH.

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 35

Fic. 2. Basidiocarps: a. Ceriporiopsis lagerheimii (Dai 12304); b. Sebipora aquosa (Dai 13592);

c, d. Tyromyces xuchilensis (Dai 12234). Scale bars: a= 2 mm; b= 2 cm; c,d = 1 cm.

SUBICULUM Generative hyphae hyaline, thin-walled, branched, flexuous,

interwoven, 3-5 um in diam.

TuBEs Generative hyphae hyaline, thin-walled, branched, flexuous,

interwoven, 3.5-4.5 um in diam. Cystidia and cystidioles absent. Basidia

barrel-shaped to pyriform, hyaline, thin-walled, with four sterigmata and

a basal clamp connection, 9-14 x 5-7 um; basidioles similar to basidia, but

slightly smaller.

Spores Basidiospores cylindrical, tapering toward apiculus, hyaline, thin-

walled, smooth, IKI-, CB-, (3-)3.2-3.7(-4) x (1.3-)1.5-1.8(-2) um, L = 3.5 um,

W= P77; Oe 2(n = 30/7).

TYPE OF ROT: White rot.

SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE: Jinghong, Xishuangbanna Nature

Reserve, Sanchahe, on rotten angiosperm wood, 7 June 2011, Dai 12304 (BJFC010586).

ECUADOR. NaPo PROVINCE, Cuyuja, 4 May 2002, Ryvarden 58240 (isotype in O).

CoMMENTs: The Chinese specimen has cream to buff pore surface after drying,

while the isotype specimen has white to cream pore surface. Otherwise, they

have same morphological characters.

ESSIEN

Fic. 3. Ceriporiopsis lagerheimii (drawn from Dai 12304).

a. Basidiospores; b. Basidia and basidioles; c. Tramal hyphae; d. Subicular hyphae.

Sebipora aquosa Miettinen, Mycol. Prog. 11: 144, 2012. Fics 2b, 4

BASIDIOME Annual, resupinate, watery and soft when fresh, becoming hard

corky when drying; <9 cm long, 5 cm wide, 8 mm thick at center. Pore surface

white when fresh, cream to yellow upon drying, more or less tallowing; pores

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 37

angular, 4-6 per mm; dissepiments thin, entire. Sterile margin cream to pale

brown, <1 mm wide. Context white to cream, soft corky, <1 mm thick. Tubes

concolorous with pore surface, hard corky, <7 mm long.

HyPHAL STRUCTURE Monomitic; generative hyphae with clamp connections,

IKI-, CB-; tissues unchanged in KOH.

SUBICULUM Generative hyphae hyaline, thin- to slightly thick-walled,

occasionally branched, more or less parallel to substrate, 3-5 um in diam.

§§o9d9

Sey ERRTe

10 pm

Fic. 4. Sebipora aquosa (drawn from Dai 13592).

a. Basidiospores; b. Tramal section; c. Subicular hyphae.

38 ... Zhao, Liu & al.

TuBEs Generative hyphae hyaline, thin- to thick-walled, unbranched,

subparallel to the tubes, 2.5-4 um in diam. Small pale-yellow resinous granules

occasionally present. Cystidia and cystidioles absent. Basidia clavate to

pyriform, with four sterigmata and a basal clamp connection, 16-24 x 5-6 um;

basidioles similar to basidia, but slightly smaller.

Spores Basidiospores cylindrical to oblong-ellipsoid, often slightly curved,

hyaline, thin-walled, smooth, usually bearing one or two guttules, IKI-, CB-,

(5.5-)6-6.5(-6.7) x (2-)2.2-2.8 um, L = 6.2 um, W = 2.5 um, Q = 2.3-2.5

(n = 60/2).

TYPE OF ROT: White rot.

SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE: Jinghong, Xishuangbannan

Nature Reserve, Wangtianshu, on fallen angiosperm trunk, 19 October 2013, Dai 13592

(BJFC015054); Nanhua County, Dazhongshan Nature Reserve, on rotten angiosperm

wood, 15 July 2013, Dai 13268 (BJFC014755).

ComMENts: The Chinese material conforms closely to the original description,

which cites slightly longer basidiospores (5.7-7.6 x 2.1-2.7 um; Miettinen &

Rajchenberg 2012).

Tyromyces xuchilensis (Murrill) Ryvarden, Mycotaxon 23: 175, 1985. Fics 2c,d, 5

BASIDIOME Annual, pileate, soft and sappy when fresh, shrinking and

becoming corky when drying. Pilei more or less semicircular, projecting

<3 cm long, 4 cm wide, and 5 mm thick at center. Pileal surface white when

fresh, becoming cream to pale ochraceous upon drying, glabrous. Pore surface

white when fresh, buff to ochraceous upon drying; sterile margin cream,

<1 mm wide; pores angular, 3-5 per mm; dissepiments thin, entire. Context

white to cream, soft corky, <1 mm thick. Tubes concolorous with pore surface,

corky, <4 mm long.

HYPHAL STRUCTURE Monomitic; generative hyphae with clamp connections,

IKI-, CB-; tissues unchanged in KOH.

CoNTEXT Generative hyphae hyaline, thin- to slightly thick-walled,

occasionally branched, interwoven, 4-7 um in diam.

TuBES Generative hyphae hyaline, thin- to slightly thick-walled,

unbranched, interwoven, more or less parallel to tubes, 3-5 um in diam.

Cystidia and cystidioles absent; basidia clavate, with four sterigmata and a basal

clamp connection, 15-18.5 x 5-7 um; basidioles similar in shape to basidia, but

slightly smaller.

Spores Basidiospores subglobose to broadly ellipsoid, hyaline, thin-walled,

smooth, IKI-, CB-, (3.3-)3.5-4.4(-4.7) « (2.3-)2.5-3.5(-3.8) um, L = 4 um,

W = 3.3 um, Q = 1.2 (n = 30/1).

TYPE OF ROT: White rot.

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 39

SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE: Puer County, Laiyanghe Forest

Park, on fallen angiosperm trunk, 6 June 2011, Dai 12234 (BJFC010517). ECUADOR.

ORELLANA PROVINCE, Yasuni National Park, 9 March 2002, Ryvarden 44669 (O).

ComMENts: The Chinese material conforms closely to the Ecuadorian specimen

and the original description, which cites slightly narrower basidiospores

(3.5-4.5 x 2.5-3 um; Ryvarden 1985).

Seer

10 um

Fic. 5. Tyromyces xuchilensis (drawn from Dai 12234).

a. Basidiospores; b. Tramal section; c. Tramal hyphae; d. Context hyphae.

Discussion

The ITS+nLSU sequence analyses revealed seven major clades of Polyporales:

antrodia clade, core polyporoid clade, fragiliporia clade, gelatoporia clade,

phlebioid clade, residual polyporoid clade, and tyromyces clade (Fic. 1). The

phylogenetic placement of the sequences from the three new Chinese records

AO ... Zhao, Liu & al.

was consistent with previous studies (Miettinen & Rajchenberg 2012; Binder

et al. 2013).

In the phylogenetic tree, Ceriporiopsis lagerheimii was closely related to

C. guidella Bernicchia & Ryvarden; however, C. guidella produces a dull yellow

to green pore surface and has larger pores (4-5 per mm) and basidiospores

(4-5 x 2.1-2.4 um; Bernicchia & Ryvarden 2003; Ryvarden & Melo 2014).

Ceriporiopsis flavilutea (Murrill) Ryvarden is similar to C. lagerheimii in

having resupinate basidiocarps with white to cream pore surface, but it has

larger pores (3-5 per mm) and ellipsoid basidiospores (3.5-4.5 x 2-2.5

um; Ryvarden 1985). Ceriporiopsis gilvescens (Bres.) Domanski resembles

C. lagerheimii by having similar basidiospores (3.5-4.5 x 1.5-2 um); however,

C. gilvescens has an orange-brown pore surface, larger pores (4-5 per mm),

and hyphae frequently covered by small, rod-like crystals (Ryvarden & Melo

2014). Ceriporiopsis lagerheimii was described from its type locality in Ecuador

(Lzessoe & Ryvarden 2010). The Chinese specimen (Dai 12304) clustered with

the isotype (O, Ryvarden 58240) with strong support. This is the first report of

C. lagerheimii from China.

Two Sebipora aquosa specimens from China (Dai 13592, Dai 13268)

clustered with the holotype (ANDA, Miettinen 8868) and two other samples

from Indonesia with strong support to form a sister lineage with Gelatoporia

subvermispora (Pilat) Niemela within the gelatoporia clade. Morphologically,

G. subvermispora differs from S. aquosa by larger pores (2-4 per mm) and

smaller allantoid basidiospores (4.5-5.5 x 1-1.5 um; Miettinen & Rajchenberg

2012; Ryvarden & Melo 2014). This is the first report of S. aquosa from China.

Two Tyromyces xuchilensis specimens (Dai 12234 from China; Ryvarden

44669 from Ecuador) formed a strongly supported lineage not in the tyromyces

clade but in the residual polyporoid clade. This unexpected result was also

noted by Binder et al. (2013). The Mexican type specimen of T’ xuchilensis

(Murrill 1171) should be reexamined in order to resolve its taxonomic status.

In the tree, T: xuchilensis appeared close to Climacocystis borealis (Fr.) Kotl. &

Pouzar. However, C. borealis is easily distinguished from T: xuchilensis by its

duplex context, larger pores (1-2 per mm), acute cystidia, larger basidiospores

(4.5-6.5 x 3-4.5 um), and habit on gymnosperms (Song et al. 2014). Tyromyces

xuchilensis was described from Mexico, and later found in Ecuador (Ryvarden

1985); this is the first report of the species from China.

Acknowledgments

We would like to express our appreciation to Prof. Yu-Cheng Dai (BJFU) for allowing

us to study his specimens. Special thanks are due to Drs. Josef Vlasak (Academy of

Sciences of the Czech Republic) and Li-Wei Zhou (Institute of Applied Ecology, Chinese

Ceriporiopsis, Sebipora & Tyromyces spp. new to China... 41

Academy of Sciences, Shenyang, China) who reviewed the manuscript. The research

was financed by the Fundamental Research Funds for the Central Universities (Project

No. 2016Z.CQ04).

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MY COTAXON

January-March 2017— Volume 132, pp. 43-62

http://dx.doi.org/10.5248/132.43

Circinella (Mucorales, Mucoromycotina) from China

RU-YONG ZHENG’, XIAO-YONG LIu* & YA-NING WANG

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

* CORRESPONDENCE TO: zhengry@im.ac.cn, liuxiaoyong@im.ac.cn

ABSTRACT—Nine taxa representing the genus Circinella—C. angarensis, C. minor var. minor,

C. minor var. asperior, C. mucoroides, C. muscae, C. nodulosa sp. nov., C. ramosa sp. nov.,

C. simplex, C. umbellata—were recorded and described or redescribed from twenty-four

different localities in seventeen cities of fifteen provinces in China. A key is provided to the

nine taxa recorded from China. No zygosporic state was formed between any pair of the

isolates studied.

Key worps—Mucoraceae, morphology, new species, taxonomy

Introduction

Circinella (van Tieghem & Le Monnier 1873) is one of the smaller genera

of the family Mucoraceae (Mucorales, Mucoromycotina). It is closely related to

Mucor but differs in forming sporangiophores with circinate branches bearing

sporangia with persistent walls rather than the non-circinate branches and

non-persistent sporangial walls characteristic of Mucor species (Hesseltine &

Fennell, 1955).

Hesseltine & Fennell (1955) recognized eight species in Circinella. Although

subsequently seven additional species were included (Faurel & Schotter 1965;

Milko 1968, 1969, 1974; Patil & Kale 1981; Arambarri & Cabello 1996), six

have since been accepted as representing other genera such as Fennellomyces,

Gongronella, Lentamyces, Mucor, and Pirella (Benny & Benjamin 1975, Benny

& Schipper 1992, Walther et al. 2013).

Six Circinella species have been described in Taiwan: C. minor, C. mucoroides,

C. muscae, C. rigida [° Mucor durus], C. simplex, and C. umbellata (Yang & Liu

1972, Xiao et al. 2013).

44 ... Zheng, Liu & Wang

In Mainland China, R.Y. Zheng isolated and identified Circinella angarensis,

C. minor, C. muscae, and C. umbellata, which had been listed (but not described

or illustrated) in Teng (1964) and Tai (1979). We collected and identified

additional Circinella strains, adding four taxa (including two new species) to

the Chinese records.

Of the nine Circinella taxa found in China described or redescribed in detail

and illustrated here, eight have been recorded from Mainland China and one

only from Taiwan.

Materials & methods

Isolation

All strains found in China were isolated from soils, dung, or rotten materials. The

Warcup (1950) dilution plate method was used for soil samples, a small part of each

dung sample was soaked in sterilized distilled water for 5-6 hours before using different

dilutions to pour plates, and rotten materials were placed directly on the media.

Cultures

The isolates of each taxon studied are listed after the description of that taxon. Our

research group isolated and identified the strains from Mainland China, and Prof.

Hsiao-Man Ho supplied those from Taiwan. Dr. K. O'Donnell kindly supplied cultures

from NRRL for comparative studies; Dr. A. Nakagiri kindly supplied culture NBRC

4453, sent by Dr. H. Naganishi. Living cultures are maintained in the Culture Collection

of the State Key Laboratory of Mycology (accession numbers prefixed with ‘C’) and

the China General Microbiological Culture Collection Center (CGMCC), and dried

cultures are deposited in the Herbarium Mycologicum Academiae Sinicae (HMAS), all

of the Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

Media & cultivation

SMA (modified synthetic Mucor agar, containing dextrose 20 g, asparagines 2 g,

KH,PO, 0.5 g, MgSO,-7H,O 0.25 g, thiamine chloride 0.5 mg, agar 20 g in 1000 mL

distilled water, pH 7) was used for morphological studies. Color codes cited in the

descriptions follow to Ridgway (1912). PDA, also adjusted to pH 7, was used for

isolation, establishing temperature-growth relationships, and mating experiments.

Cultivation period and temperature were 4-7 days at 18°C for isolating, 4-14 days at

25°C for morphological studies, 4 days at temperatures of 32-43°C for determination

of the maximum growth temperature, and 7-20 days at 18°C for mating experiments.

Results & discussion

Maximum growth temperature

The 45 living cultures studied were tested twice for their maximum growth

temperature. The eight species have a very similar maximum growth range:

(31-)36-40°C, which may reflect the close affinity among these taxa. The

number of strains is, however, too small to summarize the exact range for each

Circinella spp. nov. (China) ... 45

taxon. Since the results obtained are not distinctive enough to differentiate

one another, the maximum growth temperature is not suitable for use in the

identification of taxa in this study.

Mating experiments

Zygospores were not produced under different conditions between any of

the mating pairs.

Morphological studies

As noted for many taxa in other mucoralean genera, morphological

characteristics are found to be useful taxonomic criteria in distinguishing taxa

within Circinella.

Taxonomy

Circinella Tiegh. & G. Le Monn., Ann. Sci. Nat., Bot., 5e sér., 17: 298. 1873.

Co.Loniss fast growing, filling the Petri dish in 7-9 days at 25°C, (3-)

7-10 mm high, at first white, then blackish in one week due to formation of

sporangia; reverse occasionally wrinkled; with or without an unpleasant odor.

SPORANGIOPHORES Circinate, often in groups and borne alternately at the both

sides of the main stems of the sporangiophores. SPORANGIA borne at the tip of

the circinate branches, many-spored. COLUMELLAE well developed, variously

shaped, always with a well-defined collar. SPORANGIOSPORES numerous,

hyaline, globose to ovoid, always single-celled. CHLAMyDOspPoRES absent.

ZYGOSPORIC STATE not found. (According to Hesseltine & Fennell (1955),

zygosporangia [as “zygospores’] are nearly smooth-walled and produced

between more or less equal and unadorned suspensors on special zygophores.)

Key to Circinella taxa recorded from China

1. Sporangiospores:>10\pim-diaineter 2 i.e 8. koe bs nee bates oem weep toe C. angarensis

if, 5 POAC S Ores eNO spina amber FF ha tal arg cnt ON bale SOD Bll atte ae 2

2. Sporangiospores angular and irregular in shape ...................00. C. simplex

2. Sporangiospores neither angular nor irregular in shape ...................000. 3

3. Sporangia resistant to breaking; old colonies usually blackish grey

02,7, VSP Oe eet ee eae cet en ae C. rigida [= Mucor durus]

3. Sporangia easy to break; old colonies usually brownish .....................04. 4

4. Sterile spines present on the sporangia-forming branches...................4-. 5

4. Sterile spines absent on the sporangia-forming branches ...................06. 6

5, Apex of -columellae smooth: $0... 0005202005 604 petey taney C. minor var. minor

5 Apexcot columiellae:vernicose:s fA that etcetera! C. minor var. asperior

GaUiinbels Wwathi2=12- sporangia Wo sc Be 5 ois ow ake on abn e mn obene nm phere wptoke at C. umbellata

GxUmibels. with 4e S'sporangid Me isc. wae tan gern nee ere ees PGre REO 7

46 ... Zheng, Liu & Wang

7eopotansiophores slishthundulate: 45.6.4 9. aaah oath aad tac C. mucoroides

7. SpoTansiophOresnot UNAU Ate f cncce nares enw asm date OAR Awe es Melo ev Mine 8

8. Not forming special structures at the base of spines ................... C. muscae

8. Forming special structures. at the base of spinies. 0.5.5 ious beled Melee meals a saeled 9

9. Special structures nodulose, not branched ............. 0... ce eee ee ee C. nodulosa

9. Special structuresniot nodulose, branched). .1..3 03.43 e2.6 52. sensed C. ramosa

Fic. 1. Circinella angarensis (C43). 1. General characteristics of the sporophores; 2. Terminal

portion of the sporangiophores; 3. Columellae with small or more often large pieces of the basal

part of the sporangial wall attached;. 4. Sporangiospores.

Circinella spp. nov. (China) ... 47

25 pm

Fic. 2. Circinella angarensis (C43). More columellae with large pieces of sporangial wall attached.

Circinella angarensis (Schostak.) Zycha,

Krypt.-Fl. Brandenburg 6a: 98. 1935. FIGS 1, 2

= Mucor angarensis Schostak., Ber. Deut. Bot. Ges. 15:473. 1897.

CoLonlEs floccose, growing well at +25°C, 6.8 cm in diameter and reaching

15 mm high on PDA, but 3.5 cm in diameter and 10 mm high on SMA. OpoR

none. At first white, then Light Drab to Cinnamon Drab or Buffy Brown, reverse

Light Drab, margin irregular. Ru1zorps present. SPORANGIOPHORES arising

from both substrate and aerial mycelia, sub-straight to helicoid, main stems

48 ... Zheng, Liu & Wang

11.5-24.5 um diam., branches 6-10 um diam., brownish colored, growing

indefinitely and branching sympodially, branches with 1-2 sporangia, often

accompanied by a spine. Lateral branches circinate, single or united into

whorls and branching sympodially, branches with 1-2 sporangia, often

accompanied by a spine. SPORANGIA (53-)83-145(-160) um diam., at first

hyaline, then blackish, with spores visible inside the sporangium, deliquescing

after broken. COLUMELLAE ovoid and 61-82 x 44-60 um, broadly cylindrical

and 76 x 71 um, cylindrical and 66-98.5 x 49-70 um, nearly globose and

39-65 um diam. SPORANGIOSPORES globose and 6-11 um diam., or ovoid

and 6.5-12 x 6-9.5 um. CHLAMYDOSPORES absent. ZYGOSPORIC STATE not

formed between any pair of the (+) and (—) strains found in China under

different conditions.

STRAINS STUDIED: CHINA, BEIJING, Beijing Zoo, from dung of a special kind of pig,

30 April 1964, Zhe-fang Han (C43 = CGMCC 3.3396). USA, CALIFORNIA, from rodent

dung (C31 [= CBS 172.62 = NRRL 2628]); Los Angeles County, San Gabriel Mountains,

from dung, R.K. Benjamin (C30 [=NRRL 2410, NT of Mucor angarensis]).

Circinella angarensis, originally described as Mucor angarensis by

Schostakowitsch (1897), is recognized by having larger sporangia, columellae,

and sporangiospores than other Circinella species. Zycha (1935) transferred

this taxon to Circinella based on its original description. Hesseltine & Ellis

(1961), who re-examined its isolates, confirmed this combination and

proposed NRRL 2410 as neotype.

Circinella minor Lendn. var. minor, Bull. Herb. Boissier, 2e sér., 5: 199.1905. Fic. 3

CoLontss floccose, growing well at 25°C, 2-5(-10) mm or even higher;

at first white, then Light Drab to Cinnamon Drab or Buffy Brown, reverse

Light Drab SPORANGIOPHORES erect, main stems 8-18.5(-32) wm diam.,

branches 4-9(-14) um diam. and 59-450 um in length. SporaNGia spherical,

terminal ones (41-)57-82(-117) um diam., lateral ones 29-70(-80) um diam.

COLUMELLAE broadly ovoid, pyriform or other shape, (27-)36-63(-99) x

(20-)31-47(-68) um, applanate or hemi-spherical, (7-)13.5-45 x (9-)16-63

um. SPORANGIOSPORES spherical and 3.5-7(-9) um diam., or ovoid and

4.5-7(-11) x 3.5-6.5(-9.5) jwm,, hyaline. CHLAMYDOSPORES absent.

ZYGOSPORIC STATE not formed between any pair of the (+) and (-) strains

found in China under different conditions.

STRAINS STUDIED: CHINA, SHANxI, Datong, from dung of black bear in zoo, 21

August 1963, Fu-mei Hu (C3 = CGMCC 3.14087); from horse dung, Fu-mei Hu (C16

= CGMCC 3.14100). BEIJING, Zoological Institute, from cat dung, 3 August 1960, Hu

Fu-mei (C8 = CGMCC 3.14092). HUNAN, Hengyang City, Yueping Park, from soil, 17

Circinella spp. nov. (China) ... 49

Fic. 3. Circinella minor var. minor (C3). 1. General characteristics of the sporophores;

2. Terminal portion of the sporangiophores; 3. Columellae often with large pieces of the

basal part of the sporangial wall attached; 4. Sporangiospores.

June 1963, Ru-yong Zheng & Fei-mei Hu (C11 = CGMCC 3.14095). UNKNOWN, A.F.

Blakeslee (C32 [= NRRL 1353, = CBS 143.56]).

Circinella minor differs from other Circinella species in producing

sporangiophores that terminate in an umbel of (1-)2-4(-6) sporangia and

lacking a spine.

50 ... Zheng, Liu & Wang

Circinella minor var. asperior M.O. Reinh.,

Ber. Deutsch. Bot. Ges. 45: 131. 1927. Fic. 4

Cotonigs floccose, growing well at +25°C, (2-)3-8(-10) mm high, at

first white, becoming Old Gold at the 7th day, reverse milky white with pale

brownish tint to Pale Orange Citrine, usually with disordered patches. Odorless,

some nearly odorless. StroLons and Ruizorws absent. SPORANGIOPHORES

arising from the substrate or aerial mycelia, erect, growing indefinitely and

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Fic. 4. Circinella minor var. asperior (C1). 1. General characteristics of the sporophores; 2. Terminal

portions of the sporangiophores; 3. Columellae mostly with large pieces of the basal part of the

sporangial wall attached; 4. Sporangiospores.

Lod

Circinella spp. nov. (China) ... 51

branching sympodially, main stems (7—)11-23(-27.5) um diam., branches

(5.5-)7-9(-16) um diam., (47—)82-200(-294) um long, each branch with

a single sporangium or more often terminating into an umbel of 2-5(-6)

sporangia, hyaline to pale grayish-brown, 1-2(-4)-septate. SPORANGIA borne

on circinate branches, spherical, both terminal and lateral sporangia similar

in size, (32-)42-70(-100) um diam., dark brown color, wall translucent,

incrusted and broken. COLUMELLAE broadly ovoid or oblong-ovoid, (14-)

27-70 x (15—)23-60 um, smaller ones roundish conical, 23-46 x 23-55 um, with

1 to several apical spines, brownish to grayish brown. COLLAR always present.

SPORANGIOSPORES globose and 3.5-9(-10.5) um diam., or ovoid and 5.5-10 x

4.5-9 um. CHLAMYDOSPORES absent. ZYGOSPORIC STATE not formed between

any pair of the (+) and (-) strains found in China under different conditions.

STRAINS STUDIED: CHINA, HEBEI, Tianjin City, People’s Park, from rabbit dung,

27 May 1965, Zhe-fang Han (C1 = CGMCC 3.14085). GuizHou, Guiyang, from shrub

forest soil, 8 June 1962, Fu-mei Hu (C5 = CGMCC 3.14089). Ninex1A, Yinchuan, from

pig dung, 19 October 1964, Fu-mei Hu (C17 = CGMCC 3.14101). BE1yjING, Mentougou,

from mouse dung, 26 September 2002, Ji-qi Lu (C27 = CGMCC 3.14111). LIAONING,

Dalian, December 2013 (C42 (= M.124).

Circinella minor var. asperior differs from var. minor by having many columellae

with verrucae.

Circinella mucoroides Saito, Centralbl. Bakteriol., 2. Abth., 17: 159. 1907. FIG. 5

Cotonigs on SMA attaining 9 cm diam. in 8-9 days at 25°C, (2-)5-8

(-10) mm high, at first white, becoming Dresden Brown or Buffy Citrine

to Cinnamon Brown, Saccardos Olive after 3 weeks, Antique Brown after 2

months. Reverse Buffy Citrine or Old Gold. Odorless in most strains, rarely

with a moldy odor. STOLONs and RHIZOIDs not seen. SPORANGIOPHORES arising

from substrate or aerial mycelia, erect, growing indefinitely and branching

sympodially from substrate or aerial mycelia, erect, growing indefinitely and

branching sympodially, (70-)94-250(-505) um in length, main stems (4.5-)

9-18(-27.5) um diam., branches (3.2—)6-11(-16) um diam., each branch with

1(-2) sporangia, hyaline to pale brownish, 1-3-septate. SPORANGIA borne on

circinate branches, spherical, terminal ones (21-)35-70(-82) um, lateral ones

similar in size, dark brown color, wall translucent and incrusted, breaking and

leaving large pieces of basal collar. COLUMELLAE ovoid, cylindroid or near

pyriform and 23-55(-69) x 16-46(-55) um, globoid and (14.5-)41.5-55 um,

depressed globoid and 18.5-30 x 23-37 um. SPORANGIOSPORES globose and

3.5-7(-9) um diam, shortly ovoid and 5.0-7.5 x 4.5-6.5 um. CHLAMYDOSPORES

not seen. ZYGOSPORIC STATE not formed between any pair of the (+) and (-)

strains found in China under different conditions.

52 ... Zheng, Liu & Wang

Fic. 5. Circinella mucoroides (C6). 1. General characteristics of the sporophores; 2.Terminal

portion of the sporangiophores; 3. Columellae with large pieces of the basal part of the sporangial

wall attached; 4. Sporangiospores.

STRAINS STUDIED: CHINA, GUIZHOU, Xiuwen County, from fallow soil, 21 November

1964, Qing-tao Chen (C2 = CGMCC 3.14086). Jiancsu, Nanjing Zoo, from fox dung,

12 October 1960, Xin-sheng Li & Fu-mei Hu (C6 = CGMCC 3.14090). ZHEJIANG,

Hangzhou, Zhejiang Agricultural University, from swine dung, 4 September 1987,

Gui-qing Chen & Fu-mei Hu (C19 = CGMCC 3.14103); from hen dung, Gui-qing

Chen & Fu-mei Hu (C20 = CGMCC 3.14104). BeyinG, Changping, from vegetable

Circinella spp. nov. (China) ... 53

garden soil, 4 May 1982, Ru-yong Zheng (C23 = CGMCC 3.14107); Mentougou,

from mouse dung under a walnut tree, 26 September 2002, Ji-qi Lu (C28 = CGMCC

3.14112). YUNNAN, Xishuangbanna, from soil under a tree, 9 July 1994, Gui-qing Chen

(C26 = CGMCC 3.14110). INNER MONGOLIA, Taipushi, from mouse dung, Wei Liu

(C29 = CGMCC 3.14113). TaArwan, 26 August 2014, Xiao-man He (C45, received as

"Circinella chinensis"). UNKNOWN, A.EF. Blakeslee (C33 [= NRRL 1354]).

Circinella mucoroides is diagnosed by its slightly undulate sporangiophores,

simple sterile spines, and sporangia at the upper end of sporangiophores.

Circinella muscae (Sorokin) Berl. & De Toni, Syll. Fung. 7: 216. 1888. FIG. 6

= Circinella sydowii Lendn., Bull. Bot. Geneve, Ser. II, 5: 29. 1913.

Cotonies on SMA floccose, attaining 9 cm diam. in 8-10 days at 25°C,

4-8(-10) mm high, at first white, becoming Buffy Citrine or Saccardos

Olive in 7 days, near Walnut Brown in 10 days and Antique Brown to

Amber Brown after 2 months. Reverse Sayal Brown, with white to dirty

yellow edge surrounded. Opor absent, rarely with a faint foul smell.

STOLONS absent. Ruizorps absent or present and branching profusely.

SPORANGIOPHORES arising from substrate or aerial mycelia, erect, growing

indefinitely and branching sympodially, (6.5-)9-14(-19) um diam., branches

(37-)74-447 um in length, (4.5-)7-10 um diam.; fertile branches circinate,

with 1 or 2, sometimes 3 sporangia, mostly accompanied by a spine. SPINES

simple or dividing 1-2 times, stiff or circinate, recurved, or meandering,

(47-)70-141(-376) um long. SPoRANGIA globose to slightly dorsiventrally

flattened., 27-60(-82) um diam., hyaline or translucent when young, dark

gray to brown in age, wall persistent, breaking. COLUMELLAE variable in

shape, larger ones pyriform, oblong-ovoid, constricted or not constricted at

the middle portion, (14—)25-46 x (12—)19-32 um, smaller ones conical or

hemi-globose, (8—)12—30(-37) x (7-)11-25(-32) um, mostly hyaline, rarely

light brown, mostly smooth, a few with spines or protuberances at the apex,

often with large pieces of collars attached. SPORANGIOSPORES globose and

3.5-7(-9) um diam., ovoid and 4.5-7(-11) x 3.5-6.5(-9) um. ZYGOSPORIC

STATE not formed between any pair of the (+) and (—) strains found in China

under different conditions.

STRAINS STUDIED: CHINA, GUANGXtI, Guilin, from vegetable soil, Ru-yong Zheng (C13

= CGMCC 3.14097). HEBEI, Kailuan, from underground cable, Zu-tong Qi (C21 =

CGMCC 3.14105; C22 = CGMCC 3.14106; C24 = CGMCC 3.14108). SOUTH AFRICA,

GAUTENG, Johannesburg, from gold mine soil at a depth of 600 m and 40°C, A. Lendner

(C41 [= NRRL 1364 = CBS 107.13, type of Circinella sydowii]). UNKNOWN, (C34 [=

NRRL 1358]; C35 [= NRRL 1359]).

Circinella muscae is characterized by non-undulate sporangiophores and

simple spines often associated with sporangiophore branches. Compared

54 ... Zheng, Liu & Wang

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Fic. 6. Circinella muscae (C13). 1. General characteristics of the sporophores; 2.Terminal portion

of the sporangiophores; 3. Columellae mostly with smooth apices and with large pieces of the basal

part of the sporangial wall attached; 4. Sporangiospores.

with its allied C. mucoroides, this species produces more deeply colored

sporangiophores, fewer sterile spines, shorter and more circinate branches, and

smaller sporangia.

Circinella nodulosa R.Y. Zheng, X.Y. Liu & Y.N. Wang, sp. nov. FIG. 7

FUNGAL NAME FN570151

Circinella spp. nov. (China) ... 55

Fic. 7. Circinella nodulosa (C18). 1. General characteristics of the sporophores; 2. Terminal portion

of the sporangiophores; 3. Columellae with small or large pieces of the basal part of the sporangial

wall attached; 4. Sporangiospores.

Differs from other species of Circinella by forming unbranched nodulose structures at

the base of sporangiophores or spines.

Type: People’s Republic of China, Zhejiang Prov., Hangzhou, campus of Zhejiang

Agricultural University, from sheep dung, 4 September 1987, Gui-Qing Chen & Fu-Mei

Hu (Holotype HMAS 245380; ex-holotype culture CGMCC 3.14102 = C18).

EryMo_Locy: nodulosa, nodular, referring to the nodulose nature of the special structure

forming at the base of spines.

56 ... Zheng, Liu & Wang

CoLonies on SMA floccose, attaining 9 cm diam. in 7-9 days at 25°C, about

10 mm high, at first white, becoming Yellow Ocher to Ochraceous Tawny in

the central part, remaining white in the periphery. Reverse pale yellowish.

ODORLESS. STOLONS absent. RuIzorps abundant. SPORANGIOPHORES

arising from the substrate or aerial mycelia, erect, growing indefinitely and

branching sympodially, 59-450 um in length; main stems 9-23(-50.5) um

diam., branches 4.5-11.5(-16) um diam., each branch with 1(-2) sporangia,

1-3(-5)-septate on the branches, usually without septation on the main

stems, nodulose outgrowths may appear on the main stems or the branches

of the sporangiophores, especially at the base of the branches or spines.

SPORANGIA borne on circinate branches, globose, 26-47 um diam., light

grayish brown to pale yellowish-brown or subhyaline, breaking and leaving

a large collar. COLUMELLAE ovoid, 18.5-41.5(-50.5) x 16-32(-41.5) um,

globose to subglobose, 16-55 um diam., smaller ones depressed-globose,

9-18.5 x 14-23 um, pale to light brown. CoLLars small to large, usually

without, but sometimes with apophyses. SPORANGIOSPORES ovoid and

5.5-9(-10) x 4-7.5(-8.5) um, rarely reaching 12.5 x 9 um, globose and

(4.5-)5.5-7(-8) um diam., hyaline. ZyGOsPoRIC STATE not formed between

any pair of the (+) and (—) strains found in China under different conditions.

Circinella ramosa R.Y. Zheng, X.Y. Liu & Y.N. Wang, sp. nov. Fic. 8

FUNGAL NAME FN570150

Differs from other species of Circinella by sometimes forming, at the base of spines, a

rhizoid-like structure that branches irregularly several times or is otherwise contorted.

Type: People’s Republic of China, Inner Mongolia, from spinach field soil, June 1962,

Xun-Chu Yan (Holotype HMAS 245379; ex-holotype culture CGMCC 3.14088 = C4).

ETYMOLOGY: ramosa, from the Latin ramosus (branched), pertaining to the branched

special structure forming at the base of spines.

CoLontgs on SMA attaining 9 cm in 7-9 days at 25°C, (2-)5-10 mm high,

at first white, becoming Dark Citrine after 7 days in the central portion, or

Saccardo’s Olive after 20 days, with a white periphery which disappearing

after 3 weeks. Reverse among Aniline Yellow and Orange Citrine in 7 days,

2-5 mm high. NEAR ODORLESS. STOLONS absent. Ru1zo1ps abundant and

branching profusely at the base of the main stem. SPORANGIOPHORES arising

from the substrate or aerial mycelia, erect, growing indefinitely and branching

sympodially, curved to helicoids, (82-)141-400(-610) um in length, main

stems (8-)10-20.5(-25) um diam., branches (4.5-)7-11.5(-13) um diam.,

each branch with 1(-3) sporangia, or without sporangia and substituted by a

Circinella spp. nov. (China) ... 57

Fic. 8. Circinella ramosa (C4). 1. General characteristics of the sporophores; 2. Terminal portion

of the sporangiophores; 3. Columellae with large pieces of the basal part of the sporangial wall

attached; 4. Sporangiospores.

rhizoid-like structure that branches irregularly several times or is contorted.

SPORANGIA subhyaline, brownish to dark brown, somewhat depressed globose

or spherical, (19-)33-75(-82) um diam., broken after mature, collar large or

small. COLUMELLAE ovoid, cylindroid, rarely pyriform, (18.5-)28-64(-74)

58 ... Zheng, Liu & Wang

x (13-)30-48(-69) um, or globose and (18.5-)27.5-46(-55) um diam.,

upper most portion of the sporangiophores just beneath the sporangia often

widened and apophyses-like. SPORANGIOSPORES ovoid and 4.5-9 x 3.5-8 um,

or globose and 3.5-7 um diam., pale yellowish gray when single, gray color

in mass. ZYGOSPORIC STATE not formed between any pair of the (+) and (-)

strains found in China under different conditions.

ADDITIONAL STRAINS STUDIED: CHINA, GUANGDONG, Yingde, from soil under grasses,

4 October 1960, Ru-yong Zheng (C7 = CGMCC 3.14091). YUNNAN, Yuanjiang, from

dog dung, 21 May 1963, Zao-yuan Zhang & Yong-gui Ma (C10 = CGMCC 3.14094).

HAINAN, Chuangjiang, Cha River, from soil at river bottom, Dong-hai Ye (C14 =

CGMCC 3.14098).

Circinella simplex Tiegh., Ann. Sci. Nat., Bot., 6e sér., 1: 92 (1875). Fic. 9

Cotontges on SMA attaining 6.7 cm in 6 days at 25°C, at first white, 5-8

mm high, then submerged in the central portion, with a brownish zone

which disappear after 3 weeks, Deep Olive-Buff to Dark Olive-Buff after 20

days, 2-3 mm high. Reverse Pale Olive-Buff. Near odorless. SToLons absent.

RHIZOIDS absent. SPORANGIOPHORES arising from the substrate, erect, hyaline

to pale brownish, growing indefinitely and branching sympodially, curved,

(5-)8-12.5(-16) um diam., branches slightly widening near the basal parts,

(13.5-)31-68(-114) um in length, each branch with one sporangium, branches

(0-)1(-2)-septate, spines absent. SPORANGIA spherical, (27.5-)32-61.5(-69)

um diam., at first hyaline, then brown, deliquescing after broken. Collar small

or absent. COLUMELLAE variable in shape, broadly ovoid, cylindroid or near

pyriform, constricted or notconstrictedatthe middle portion and (11-)14.5-37.5

(-44.5) x (7-)12.5-27.5(-41) um, depressed globoid to conicaland (5.5-)12.5-23

(-33) x (7-)14-25(-41) um, mostly hyaline, the larger ones light brown, mostly

smooth, a few with 1 to several spines at the apex. SPORANGIOSPORES very

irregular in shape, mostly 3.5-9 x 2-7 um, larger ones 10-16.5 x 4-5 um diam.,

with conspicuous oil droplets, hyaline. CHLAMyDosporEs nearly spherical,

16-29 um diam., each containing one larger or several small droplets, elongated

on substrate mycelia. ZyYGOsPORIC STATE not formed.

STRAINS STUDIED: CHINA, TAIWAN, Taibei, from soil (C44 [= BCRC FU30085]).

BRAZIL, from soil (C46 [= BCRC 31711 = CBS 142.35 = IFO 6412 = IMI 252490 =

NRRL 2407)).

Circinella simplex has the typical morphological characteristics of this genus,

circinate branches bearing sporangia with persistent walls, and was recognized

by producing irregular sporangiospores.

Circinella spp. nov. (China) ... 59

i 25 pm

[aaa

Fic. 9. Circinella simplex (C44). 1. General characteristics of the sporophores; 2. Terminal portion

of the sporangiophores; 3. Columellae with small or large pieces of the basal part of the sporangial

wall attached; 4. Sporangiospores; 5. Chlamydospores.

60 ... Zheng, Liu & Wang

Circinella umbellata Tiegh. & G. Le Monn.,

Ann. Sci. Nat., 5e sér., 17: 300. 1873. FIG. 10

= “Circinella chinensis” H. Nagan. &Kojiro, J. Ferm. Tech. 20: 409. Pl. 3. 1942, nom. inval.

Cotonizes on SMA attaining 9 cm diam. in 7-9 days at 25°C, (3-)7-10

mm high, at first white, Old Gold or Isabella Color in 7 days, Light Brownish

Olive to Cinnamon Brown in 2 weeks, then Hazel to Antique Brown after

2 months. Reverse similar to the obverse side or dirty grayish brown.

STOLONS and RuIzorps absent. SPORANGIOPHORES arising from the substrate

or aerial mycelia, erect, growing indefinitely and branching sympodially,

(82—)130-258(-282) um in length, main stems (9-)14-27.5(-33) um diam.,

branches (5.5—)7-14(-18.5) um diam., each branch with a single sporangium

or more often terminating into an umbel of 2-12 SPORANGIA borne on circinate

branches which are hyaline to somewhat brownish and (0-)1-2-septate,

spherical, (27—)41-94(-118) um diam., dark brown color, wall translucent,

incrusted and breaking. COLUMELLAE ovoid, oblong-cylindroid, or pyriform

and (23-)30-60(-74) x (18.5-)24-46(-64.5) um, or globose and (18.5-)

27.5-50.5(-69) um diam. in the larger ones, depressed-globose to conical and

(9-)18.5-41.5(-69) x (14—)23-46(-59) um in the smaller ones, mostly smooth,

a few with 1 to several spines at the apex, brownish or light grayish-brown.

SPORANGIOSPORES globose, (4.5—)5.5-10 um diam., or ovoid, (6—)7—11(-12.5)

x (5.5-)6.5-10(-11) um, easy to germinate and becoming irregular in shape.

ZYGOSPORIC STATE not formed.

STRAINS STUDIED: CHINA, BEeyinG, Zhongguancun, from mouse dung, 8 June 1961,

Fu-mei Hu (C9 = CGMCC 14093). HuNAN, Hengyang, Yueping Park, from soil, 17

June 1963, Ru-yong Zheng & Fu-mei Hu (C12 = CGMCC 3.14096). INNER MONGOLIA,

Arshaan, Yiershi, from wilted Rhododendron flower, 23 June 1963, Xi-ling Chen (C15 =

CGMCC 3.14099). HAINAN, Limu Mountain, from soil under jackfruit plant, Song Lin

(C25 = CGMCC 3.14109). JAPAN (C40 [= NBRC 4453 = CBS 140.28, type of "Circinella

chinensis"]). UNKNOWN, A.E. Blakeslee (C36 [= NRRL 1351 = CBS 144.56]; from dog

dung (C37 [= NRRL 1366 = CBS 145.56]).

Circinella umbellata, the type species for Circinella, differs from other species

in the genus in its sporangiophores that terminate in an umbel of 2-12

sporangia. Circinella “chinensis”, which was published by Naganishi & Kojiro

(1942) without a Latin diagnosis, was treated by Hesseltine & Fennell (1955)

as a synonym of C. mucoroides. Naganishi (1974), however, pointed out

sporangiophore differences by comparing the Osaka strain (IFO 4455) of

C. mucoroides and Dairen strain (NRRL 1354) of C. chinensis. Our study of the

ex-type of C. “chinensis” reveals that morphologically it belongs to C. umbellata.

Circinella spp. nov. (China) ... 61

Fic. 10. Circinella umbellata (C9). 1. General characteristics of the sporophores; 2.Terminal portion

of the sporangiophores; 3. Columellae usually with large pieces of the basal part of the sporangial

wall attached; 4. Sporangiospores.

Acknowledgments

The project was partially supported by the National Science Foundation of China

(No. 31370068) and the Foundation of the Knowledge Innovation Program of the

Chinese Academy of Sciences (No. KSCX2-EW-J-6). Cultures supplied by NRRL and

NBRC used in this study are gratefully acknowledged. Many friends and colleagues

62 ... Zheng, Liu & Wang

who supplied us with their samples or cultures are greatly appreciated. We also thank

Ms Xiang-fei Zhu and Ms Hong-mei Liu (both of this laboratory) for respectively inking

the line drawings and tending the Circinella strains kept in the laboratory. Ms Wen-hua

He is thanked for her devotion to the taking care of our Circinella strains kept in the

CGMCC Culture Collection for many years. Special thanks are due to Dr. P.M. Kirk

and Dr. G.L. Benny for serving as presubmission reviewers of this paper and for their

valuable comments to improve the manuscript.

Literature cited

Arambarri AM, Cabello MN. 1996. Circinella lacrymispora sp. nov. A new mucoral isolated from

Argentine soils. Mycotaxon 57: 145-149.

Benny GL, Benjamin RK. 1975. Observations on Thamnidiaceae (Mucorales). New taxa, new

combinations, and notes on selected species. Aliso 8(3): 301-351.

Benny GL, Schipper MAA. 1992. Observations on Thamnidiaceae (Mucorales). IV. Pirella.

Mycologia 84: 52-65. http://dx.doi.org/10.2307/3760401

Faurel L, Schotter G. 1965. Notes mycologiques VI. Sur quelques champignons coprophiles

d'Afrique Equatoriale. Cah. La Maboke 3: 123-133.

Hesseltine CW, Fennell DI. 1955. The genus Circinella. Mycologia 47: 193-212.

http://dx.doi.org/10.2307/3755410

Milko AA. 1968. [The nomenclature of some Mucorales with diagnostic keys to species of Circinella,

Thamnidium and Kickxella.| Novosti Sist. Nizsh. Rast. 5: 79-88. [In Russian]

Milko AA. 1969. [A new species of the genus Circinella van Tieghem et Le Monnier.] Novosti Sist.

Nizsh. Rast.6: 96-102. [In Russian]

Milko AA. 1974. Opredeltiel mukoral nykh gribov. Naukova Dumka, Kiev. 303 p. [In Russian]

Naganishi H. 1974. On Circinella species isolated from Korean yeast cake (Kyokusi). Trans. Mycol.

Soc. Japan 15:175-177.

Naganishi H, Kojiro K. 1942. Circinella. Jour. Ferm. Tech. 20:408-414. [In Japanese]

Patil SD, Kale JC. 1981. A new species of Circinella van Tiegh. and Le Monn. Current Science

50(12): 544.

Ridgway R. 1912. Color standards and nomenclature. The Author, Washington D.C. 44 pp.,

53 colored plates.

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press. 1527 p.

Teng SC. 1964. Fungi of China. [RP Korf (ed.)]. Mycotaxon Ltd., Ithaca, New York. 586 p.

van Tieghem P, Le Monnier G. 1873. Recherches sur les Mucorinées. Ann. Sci. Nat., Bot. 5e sér.,

17: 261-399,

Walther G, Pawtowska J, Alastruey-Izquierdo A, Wrzosek M, Rodriguez-Tudela JL, Dolatabadi S,

Chakrabarti A, de Hoog GS. 2013. DNA barcoding in Mucorales: an inventory of biodiversity.

Persoonia 30: 11-47. http://dx.doi.org/10.3767/003158513X665070

Warcup JH. 1950. The soil-plate method for isolation of fungi from soil. Nature 166: 117-118.

http://dx.doi.org/10.1038/166117b0

Xiao BX, Ho HM, Liu GY. 2013. Studies on the taxonomic and phylogenetic relationship of the

genus Circinella (Mucorales) from Taiwan Prov. of China. p. 13, in: Eleventh Cross-Taiwan

Straits Joint Symposium for Mycology, 17-19 August, 2013. Beijing. [In Chinese; this citation

translated into English by RY Zheng]

Yang BY, Liu CH. 1972. Preliminary studies on Taiwan Mucorales (1). Taiwania 17(3): 293-303.

Zycha H. 1935. Mucorineae. In Kryptogamenflora d. Mark Brandenburg 6a: 1-264.

MY COTAXON

January-March 2017— Volume 132, pp. 63-72

http://dx.doi.org/10.5248/132.63

Astraeus ryoocheoninii sp. nov. from Korea and Japan

and phylogenetic relationships within Astraeus

RuHIM Ryoo!,», HONG-DUCK Sou’, HYUN PARK? & KANG-HYEON Ka!

"Division of Wood Chemistry and Microbiology, National Institute of Forest Science,

Seoul 02455, Republic of Korea

? Graduate School of Agricultural and Life Sciences, The University of Tokyo,

Tokyo 1138657, Japan

° Division of Global Forestry, National Institute of Forest Science,

Seoul 02455, Republic of Korea

“ CORRESPONDENCE TO: rryoo@korea.kr

ABSTRACT— The morphological characteristics of a collection by C.I. Ryoo and K.-H. Ka

from the demilitarized zone (DMZ) of Korea, invalidly published as “Astraeus koreana’, are

re-evaluated, described, and illustrated. Phylogenetic analysis using nrITS sequence data

indicate that “Astraeus koreana’, an unnamed “Japanese Astraeus group 2”, and a Japanese

specimen E0059827 all differ from A. hygrometricus var. koreanus and represent a new

species, proposed here as Astraeus ryoocheoninii.

Key worps—basidiospore, ornamentations, peridium, ectomycorrhiza

Introduction

Species of Astraeus are ectomycorrhizal and form symbiotic associations

with various tree species in the genera Alnus, Castanea, Eucalyptus, Pinus, and

Pseudotsuga (Malajczuk et al. 1982, Molina 1979, Nouhra & Dominguez 1998,

Trappe 1967). Mycorrhizal synthesis studies have been conducted to confirm

their host specificity in the field (Danielson 1984; Petcharat 2005; Molina 1979,

1981; Molina & Trappe 1982). Mycorrhizae carry out the efficient recycling of

nutrients between roots of host plant and fungi in soil (Malajczuk et al. 1982).

Astraeus species are also economically important as they are collected for sale

in commercial markets as edible mushrooms (Maiti et al. 2008).

64 ... Ryoo & al.

Astraeus (Sclerodermatineae, Boletales, Agaricomycetes) was originally

described by Morgan (1889). The genus contains seven widely distributed

species (http://www.mycobank.org) and has been recorded from Australasia,

Africa, Asia, Europe, and North and South America (Coker & Couch 1928,

Cunningham 1944, Dring 1964, Imazeki & Hongo 1989, Liu 1984, Nouhra

& Dominguez 1998, Wilson et al. 2012). The type species, A. hygrometricus

(Pers.) Morgan, was originally described in Geastrum; a Korean variety has

been described as A. hygrometricus var. koreanus VJ. Stanék [= A. koreanus

(VJ. Stanék) Kreisel] (Stanék 1958, Kreisel 1976). Several new Astraeus species

have been described recently from Asia: A. asiaticus Phosri et al., A. odoratus

Phosri et al. [= A. thailandicus Petcharat], and A. sirindhorniae Watling et al.

(Petcharat 2005; Phosri et al. 2004, 2007, 2014). Recently Astraeus hygrometricus

var. koreanus and an unnamed Astraeus taxon from Japan (“Clade VII,

Japanese Astraeus group 2” of Fangfuk et al. 2010) were morphologically and

molecularly re-evaluated (Fangfuk et al. 2010). However, their relationship

was left unresolved, and Fangfuk et al. (2010) concluded that further studies

were necessary to determine the phylogenetic position of var. koreanus and the

identification of “Japanese Astraeus group 2.

Until 2001, Astraeus was largely defined by two species, A. hygrometricus and

A. pteridis (Kirk et al. 2001, Zeller 1948). Since then several new species have

been described using morphological characteristics and molecular methods

(Phosri et al. 2004, 2007; Fangfuk et al. 2010). Astraeus asiaticus and A. odoratus

were described as phylogenetically distinct from A. pteridis. SEM was used to

characterize basidiospore ornamentation as a character in a taxonomic key of

the genus. Phosri et al. (2007) suggested that the A. hygrometricus complex forms

a polyphyletic group in ITS rDNA sequence analyses. These studies showed

that in Astraeus phylogenetic clades largely correlate with geographical areas.

Fangfuk et al. (2010) also confirmed polyphyletic relationships within Astraeus.

From 1995 to 2000, researchers from NIFoS (National Institute of Forest

Science) conducted an ecological investigation in the Korean demilitarized zone

(DMZ) including the civilian control zone (CCZ). In 1998, an undetermined

Astraeus species was collected from the DMZ, described, and photographed by

C.I. Ryoo and K.-H. Ka. This collection was named “A. koreana” by Ryoo et al.

(1999), but the name was invalid because no Latin description was included and

no type specimen was designated. “Astraeus koreana” is macro-morphologically

similar to A. hygrometricus var. koreanus (Stanék 1958) and shares micro-

morphological characteristics with “Japanese Astraeus group 2” (Fangfuk et al.

2010).

Our study taxonomically re-evaluated A. hygrometricus var. koreanus,

“A. koreana’, and “Japanese Astraeus group 2” by using phylogenetic analysis to

Astraeus ryoocheoninii sp. nov. (Korea) ... 65

identify which morphological characteristics are important for distinguishing

taxonomic boundaries. Astraeus ryoocheoninii is proposed as a new species

incorporating “Astraeus koreana” and “Japanese Astraeus group 2”.

Materials & methods

Morphological characteristics

We studied the macro-morphological characteristics of fresh material as recorded

and photographed in 1998 by C.I. Ryoo & K.-H. Ka and their dried material conserved

in the herbarium of National Institute of Forest Science, Hongnung Arboretum,

Seoul, the Republic of Korea (KFI). Micro-morphological features were described

from dried materials mounted in 3% KOH, Melzer’s reagent, and cotton blue reagent

under a Leica DM 2500 microscope using DIC optics. Basidiospore ornamentation

was examined under a Hitachi S-3500N electron microscope using samples coated in

platinum palladium at 10 kV.

Molecular analysis

DNA was extracted from dried material using the CTAB (cetyltrimethyl ammonium

bromide) buffer method of Rogers & Bendich (1994). The rDNA ITS region was

amplified by PCR using primers ITS1 and ITS4 (White et al. 1990). A 50 ul total volume

per PCR reaction contained template DNA (1.2 ul), 10x buffer, 0.5 M KCI, 0.1 M Tris-

HCl, 0.1 % Triton X-100, and 15 mM MgCl, 2.5 mM dNTP, 100 pM of each primer,

and Taq polymerase (5 unit/ul). PCR thermal cycler protocols comprised 35 cycles of

30 sec at 94 °C, 30 sec at 56 °C, and 1 min at 72 °C with the first denaturation and last

extension times extended to 5 min at 72 °C (Gardes & Bruns 1993). Cycle sequencing

was performed using the primers ITS1 and ITS4 using BigDye™ cycle sequencing kit,

version 3.1 (Applied Biosystems). Purified DNAs were directly sequenced on an ABI

Prism TM 377 DNA automatic DNA Sequencer (Applied Biosystems).

Phylogenetic analysis

A total of 28 sequences, one newly extracted in this study and 27 retrieved from

GenBank, were aligned using Clustal X (Thompson et al. 1997) implemented in the

program MEGA v 5.05 (Tamura et al. 2011). Parsimony analyses were performed in

PAUP’ v 4.0b10 (Swofford 2002) with bootstrap statistics generated using a heuristic

search with 1000 replicates, each with 10 random sequence additions, and TBR

branch swapping. Bayesian analyses were performed in MrBayes v 3.1 (Ronquist

& Huelsenbeck 2003) using 2,000,000 generations and 4 chains under the general

time reversible (GTR) model with gamma-distributed substitution rates. Trees

were saved every 100th generation, and the first 1000 of 2001 trees were discarded

as the burn-in. A 50% majority rule consensus tree on treefiles from Bayesian and

parsimony bootstrap analyses was used to compute posterior probabilities (PPs) and

bootstrapping (BS) values respectively. The outgroup of Scleroderma verrucosum

(Genbank AJ629886) and Pisolithus sp. (AJ629887) was used in the systematic dataset

following the methods by Phosri et al. (2007). Sequences used in this study are listed

in TABLE l.

66 ... Ryoo & al.

TABLE 1. List of Astraeus, Pisolithus, and Scleroderma sequences

used in the phylogenetic analysis

TAXON COUNTRY HOsT PLANT GENBANK NO.

A. asiaticus Thailand > AJ629381

Thailand — AJ629382

Thailand — AJ629386

A. hygrometricus USA _ AJ629398

USA — AJ629399

USA — AJ629402

USA — AJ629403

Greece — AJ629404

Spain => AJ629408

A. hygrometricus var. koreanus Japan Pinus thunbergii AB535105

Japan Pinus thunbergii AB535106

Japan Pinus thunbergii AB535107

A. odoratus Thailand Dipterocarpaceae AB507406

Thailand — AB535108

Thailand Dipterocarpaceae AB535113

Thailand Dipterocarpaceae AJ629875

Thailand — AJ629880

papierias USA = AJ629409

USA = AJ629410

A. ryoocheoninii Korea Pinus densiflora KC985146

Japan — AB507397

Japan = AB535109

Japan Pinus densiflora AB535110

Japan Quercus serrata AB535111

Japan Quercus sp. AB535112

Japan Carpinus laxiflora AJ629405

PISDIBIHISSD. Thailand = AJ629887

S. verrucosum Spain A AJ629886

Results

Phylogenetic analysis

Based on nrITS sequence data obtained in this study and from GenBank,

phylogenetic placement and relationships of Astraeus taxa were inferred

from the Bayesian and MP analyses. No difference was found between the

topologies using different methodologies so only the results from the Bayesian

analysis is shown (Fic. 1). The aligned dataset included 595 total characters,

with 393 constant, 85 variable, and 117 parsimony informative in MP analysis.

This dataset produced 1,000 trees with 315 steps of tree length, a consistency

Astraeus ryoocheoninii sp. nov. (Korea) ... 67

index (CI) of 0.8349, a homoplasy index (HI) of 0.1651, and a retention index

(RI) of 0.9091.

Five Astraeus clades are recognized in Bayesian and parsimony analyses

(Fic. 1). Clade A, representing A. hygrometricus, is subdivided into three

geographical subclades: North American (0.81 PP, 0.76% BS); South

European (0.92 PP, 91% BS); and Japanese (0.80 PP, 83% BS); the Japanese

subclade represents A. hygrometricus var. koreana, which is confirmed as a

variety rather than a species as suggested by Kreisel (1976). Clade B, here

named A. ryoocheoninii, includes “A. koreana’, “Japanese Astraeus group 2”,

AJ629399 USA

0.81/ 76} AJ629402 USA North

AJ629398 USA PL ie a

0.58 AJ629403 USA

1.007 AB535106 Japan ‘| Japan pret We

0.90/84 /100+AB535105 Japan | A. hygrometricus hygrometricus

| 44 AB535107 Japan var. koreanus

AJ629404 Greece South

0.92/91-—~ AJ629408 Spain | Europe

0.94/90 AB535110 Japan

0.56 KC985146 Korea

0.92/85 AB535111 Japan Sdeke

: AB535112 Japan sya

A. ryoocheoninii

1.00/99 AB535109 Japan

AB507397 Japan

AJ629405 Japan

oew7t giiceadtnaina

alan

LOOMO0! AB507406 Thailand | Clade/C

AB535113 Thailand | 4: 0doratus

BeuoS AJ629875 Thailand

0.69/ 72r AJ629382 Thailand

1.00/ 100] \— A J629386 Thailand Clade/D

AJ629381 Thailand Aig eItHGHS

1.00/98 AJ629409 USA Clade/E

AJ629410 USA A. pteridis

Scleroderma verrucosum AJ629886

Pisolithus sp. AJ629887

0.1

Fic. 1. Phylogenetic analysis for the genus Astraeus based on (ITS1+5.8S+ITS2) rDNA region.

Bayesian consensus tree used to a 50% majority-rule carried out two million generations and four

chains. Maximum parsimony analysis generated from 1000 replicates with 10 random addition

sequences. Because no differences were found between the tree topologies from the two analyses,

only the MCMC tree is shown. Numbers at each branch indicate posterior probability (left) and

bootstrap support (right).

68 ... Ryoo & al.

and Japanese specimen E00159827 (GenBank AJ629405) in a monophyletic

clade (1.00 PP, 99% BS). The other three monophyletic clades represent: clade

C—A. odoratus (1.00 PP, 100% BS); clade D—A. asiaticus (1.00 PP, 100% BS);

and clade E—A. pteridis (1.00 PP, 98% BS).

Taxonomy

Astraeus ryoocheoninii Ryoo sp. nov. FIG. 2

MycoBank MB 804156

“Astraeus koreana” C.I. Ryoo, KMS Newsletter 11(2): 20, 1999, nom. inval.

“Japanese Astraeus group 2” Fangfuk et al., Mycoscience 51: 291-299, 2010.

Differs from Astraeus hygrometricus var. koreanus by its larger basidiomes and smaller

basidiospores.

TypE—The Republic of Korea, Central and Eastern mountainous region near the DMZ

of Korea, Kosung, Mt. Hangrohbong, alt. + 850 m, on sandy soil associated with Pinus

densiflora, 20 July 1998, C.I. Ryoo & K.-H. Ka (Holotype, KFI-DMZ002 ; GenBank

KC985146).

EryMoLoGy—Named in honour of mycologist, Dr. Cheon-In Ryoo.

BASIDIOME globose, rough, with an ostiole encircled by a cracked peridium

60-100 mm wide when mature; the exoperidium splitting longitudinally

and re-curving into star-like rays, expanding when exposed to moisture and

re-coiling towards gleba upon desiccation. ExOPERIDIUM slightly viscous,

smooth, grayish brown, covering endoperidium, splitting into 15-19(-21) little,

star-like rays that open when hydrated, upright at first, curving back, inner

surface becoming distinctly cracked with irregular trapezoid and rhomboid

patterns, closing in dry weather. ENDOPERIDIUM a thin membranous layer of

tissue, enclosing the spore mass in a 20-40 mm diameter sac, nearly round, with

an irregularly shaped ostiole at top, surface felty rough, whitish, becoming gray

to brown. BAsIpIospoRE mass whitish when young, brownish and powdery

at maturity. BastpiosporEs globose, (5.3-)7.0-9.1(-9.5) tm in diameter

(n = 48), (4.8-)5.0-9.0 um in diameter without the ornamentation, chocolate

dark brown, cell wall thickened, with spiny, moderately dense ornamentation

with rounded (sometime spiked) top, (0.5—)0.6-1.3(-1.5) um long, dark brown,

negative reaction from 3% KOH, Melzer’s reagent (inamyloid) and cotton blue

reagent. PARACAPILLITIUM hyphae (4.0—)5.0-6.0(-6.5) tum in diameter, septate,

with clamp-connection-like structures present on some septa.

ORIGINAL DESCRIPTION OF “ASTRAEUS KOREANA” (Ryoo et al. 1999): “These

specimens were characterized by having the inner surface of the exoperidium

distinctly cracked in irregular patterns, numerous, 15-19(-21) rays and 5.7-9.3

lum average size of basidiospores with short, moderately dense ornamentation,

growing symbiotic in association with Pinus densiflora. Korean common name of

these taxa were called as Seonbi-meonji-beoseos.”

Astraeus ryoocheoninii sp. nov. (Korea) ... 69

Fic. 2. Astraeus ryoocheoninii (KFI-DMZ002, holotype). A. basidiomata; B. basidiospores under

DIC microscope; C. basidiospores under SEM. Scale bars: B = 10 um; C = 5 um

ADDITIONAL SPECIMENS—Japanese specimens now included in A. ryoocheoninii were

listed and described in Phosri et al. (2007, as ASTRAE 94 = E00159827, GenBank

AJ629405), and in Fangfuk et al. (2010, as “Japanese Astraeus group 2”).

ComMENTS — The Japanese specimens of Astraeus ryoocheoninii agree with

the Korean holotype description in basidiospore size and dense ornamentation

with short spines (TABLE 2).

70 ... Ryoo & al.

TABLE 2. Comparison of taxonomic characteristics of Astraeus hygrometricus and

Asian Astraeus species

ACUTE SPORE SIZE

TAXON SPORE ORNAMENTATION CLADE

RAYS (uum)

A. hygrometricus 6-15" 5.2-13.9° Dense, narrow, 0.7-1.3 um? A

a ;

» oe A ae 15-227 -7.6-12.9" _ Tightly dense, narrow, 0.8-1.6 wm* A

A. ryoocheoninii -

15-21 3-9, lightl , 0.6-1. B

Ba ea ni 5 5.3-9.5 Slightly dense, 0.6-1.3 um

A bari

oe ee ” 5-13? 5.2-9.6° Moderately dense, 0.6-1.7 um? B

Astraeus group 2

A. ryoocheoninii -

— .8-9.0" Dense, 0.8-1. 7 B

AJ629405 6.8-9.0 ense, 0.8-1.6 um

A. odoratus 3-9° 7.5-15.2° Moderately dense, narrow, 1.0-1.7 um‘ C

A. asiaticus 5-12? 8.8-15.2° Very dense, 0.9-1.45 um? D

* Fangfuk et al. (2010); > Phosri et al. (2007); * Phosri et al. (2004).

The A. ryoocheoninii holotype is similar to A. hygrometricus var. koreanus in

the high number of split acute rays, but A. hygrometricus var. koreanus differs

by its larger basidiospores with longer, denser ornamentation (TABLE 2),

much smaller basidiocarps (15-20 mm), and much thinner endoperidium

(10-15 mm; Stanék 1958; Imazeki & Hongo 1989; Fangfuk et al. 2010). The

type locality of A. ryoocheoninii was a warm, dry locality associated with

stands of Korean red pine (Pinus densiflora) on sandy soil, whereas the type

locality of A. hygrometricus var. koreanus was a stony plateau with an eastern

orientation among dwarf-oaks and scattered pines in the mountains of North

Korea (Stanék 1958: 632, 819).

The two Thai species, A. odoratus and A. asiaticus, have larger basidiospores

and fewer cracked rays than A. ryoocheoninii (TABLE 2). The basidiospore

spines of A. odoratus are narrower, more coalescent, and less densely arranged

than those of A. ryoocheoninii (Phosri et al. 2004, 2007; TABLE 2).

Discussion

The studies of Phosri et al. (2007) and Fangfuk et al. (2010) resolved

several questions concerning the phylogenetic relationships of the Astraeus

species. The present study uses ITS rDNA variation to establish the systematic

relationship among A. hygrometricus var. koreanus, “A. koreana’, and

“Japanese Astraeus group 2’, and to identify morphological characteristics

Astraeus ryoocheoninii sp. nov. (Korea) ... 71

that can be used to identify these species. This study suggests that Astraeus

hygrometricus var. koreanus should be recognized at the rank of variety within

the A. hygrometricus complex, rather than at specific rank as proposed by

Kreisel (1976). The new species Astraeus ryoocheoninii is described for the

well-supported clade that includes the sequences of “A. koreana’, “Japanese

Astraeus group 2”, and Japanese specimen E00159827 (GenBank AJ629405),

previously loosely associated with A. hygrometricus by Phosri et al. 2007 and

with “Japanese Astraeus group 1” by Fangfuk et al. 2010).

Acknowledgments

This paper is dedicated to the memory of Dr. Cheon-In Ryoo (1951-2009). The

authors appreciate Vladimir Antonin (Moravian Museum, Czech Republic) for his

comments about the nomenclature. We wish to thank our reviewers, Dr. Andrew W.

Wilson (Purdue University, USA) for revising the manuscript and valuable comments

and Dr. Mikael Jeppson (Sweden) and Dr. Ronald H. Petersen (University of Tennessee,

USA) for critical remarks. This work was supported by project (FP 0801-2010-01) of

National Institute of Forest Science.

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MY COTAXON

January-March 2017— Volume 132, pp. 73-78

http://dx.doi.org/10.5248/132.73

New records of Caloplaca, Hydropunctaria, and Verrucaria

from Turkey and Asia

KADIR KINALIOGLU

Department of Biology, Faculty of Science and Arts, Giresun University,

Giire Yerleskesi, Giresun, Turkey

CORRESPONDENCE: kkinalioglu@hotmail.com

ABsTRACT— Three lichen species—Caloplaca oleicola, Hydropunctaria adriatica, and Verrucaria

elaeina—are described as new to Turkey and to Asia.

Key worps—Ascomycota, Corylus sp., lichenized fungi, Teloschistaceae, Verrucariaceae

Introduction

The lichen biota of Turkey has received considerable attention in recent

years. However, additional and more intensive lichenological field studies

are needed for a complete lichen checklist of Turkey, because some regions

of Turkey have not been investigated. Additional lichen records for Turkey

are reported almost every year (e.g. Oran & Oztiirk 2010, Arslan et al. 2011,

Karagoz et al. 2011, Kinalioglu & Aptroot 2011, Vondrak et al. 2012, Aslan

& Yazici 2013, Cobano§glu et al. 2013, Yazici & Aptroot 2015, Senkardesler et

al. 2016), including species new to science (e.g., Aptroot & Yazici 2012). In

this paper we describe three lichen species: Caloplaca oleicola, Hydropunctaria

adriatica, and Verrucaria elaeina, as new records for Turkey and Asia.

Materials & methods

The specimens examined are deposited at the herbarium of the Biology Department,

Faculty of Science and Arts, Giresun University, Giresun, Turkey (GUB). They were

studied with standard anatomical and morphological methods with the help of

traditional chemical reagents (K, C, P, and I) used in lichenology (Smith et al. 2009).

74 ... Kinalioglu

rea

5 ol

™ '

a a

WN |

Fic. 1. Caloplaca oleicola (GUB 2387): habit. Scale bar = 1 mm.

Taxonomy

Caloplaca oleicola (J. Steiner) van den Boom & Breuss,

Mycotaxon 56: 131. 1995. Fig. 1

Thallus thin, white or pale grey, smooth to very slightly rimose. Apothecia

black, 0.3-0.7 mm diam., usually scattered, without a thalline margin, proper

margin persistent, well-delimited and mostly glossy, disc slightly concave to

flat. Epihymenium dark brown. Ascospores colourless, 10-15 x 5.5-9 um,

ellipsoid, septum 3.5-5 um wide. Thallus C-, K-, KC-, PD-; apothecial proper

margin and epithecium C-, K+violet-red, KC-, PD-.

SPECIMEN EXAMINED: TURKEY. Grresun: Piraziz, Aydere village, 40°56’33”N

38°06’46”E, 156 m, on Corylus sp., 30 March 2006, leg. K.Kinalioglu, det. A.

Khodosovtsev (GUB 2387).

RemMARKs—Our Turkish collection had apothecia and ascospores of a similar

size to those described in the Caloplaca oleicola protologue (apothecia <0.6 mm

diam, ascospores 10-16 x 6-8 um with 5 um septum; Boom & Etayo 1995).

Caloplaca oleicola is an uncommon corticolous species with black apothecia,

previously found only on bark of Alnus incana and Olea europaea in Europe

and North America (Boom & Etayo 1995, Goward et al. 1996); but in Turkey,

we collected it from smooth bark of Corylus sp. in a shady and humid hazelnut

garden. ‘This is the first report of the species from Asia.

Caloplaca oleicola is clearly distinguished from C. servitiana by its thin white

thallus and lack of a thalline exciple (Vondrak et al. 2010).

New Caloplaca, Hydropunctaria, and Verrucaria spp. for Turkey & Asia... 75

Fic. 2. Hydropunctaria adriatica (GUB 2388): habit. Scale bar = 1 mm.

Hydropunctaria adriatica (Zahlbr.) C. Keller & Gueidan,

Taxon 58(1): 194. 2009. Fic. 2

Thallus epilithic, superficial, shiny black. Perithecia scarce, prominent,

concolorous with thallus, globose, 0.3-0.5 mm in diam. Ascospores ovoid,

oblong, 12.5-17.5 x 6-8.75 um. Pycnidia not observed. Thallus and apothecia

C-, K-, KC-, PD-.

SPECIMEN EXAMINED: TURKEY. GrrESUN: Kesap, Degirmenagzi village, sea shore

(mostly intertidal zone), 40°58’20’N 38°37’23”E, 1 m, on siliceous rock, 11 April 2010,

leg. K.Kinalioglu, det. A. Orange (GUB 2388).

REMARKS— Our Turkish sample is distinguished from the European samples by

its substrate preference, its smaller apothecia, and somewhat bigger ascospores;

European material had apothecia 0.45—1 mm in diam and ascospores 12-15 x

7.5-8.5 um (Zschacke 1933-34).

76 ... Kinalioglu

This poorly known species was previously found on calcareous rocks splashed

by seawater on the shores of the Adriatic, Aegean, Black, and Mediterranean

Seas (Zschacke 1933-34, Rechinger 1951, Orange 2012). It is common in the

Adriatic and Aegean Seas where it forms the so-called “black band” on seashore

rocks (Rechinger 1951). In Turkey, it grows mostly on intertidal siliceous rocks

on the Black Sea shoreline. This is the first report of the species from Asia.

Hydropunctaria adriatica differs from H. maura usually by its thinner

thallus, lack of a dark basal layer, and exciple unpigmented below (Zschacke

1933-3), Orange 2012).

Verrucaria elaeina Borrer, in Smith & Sowerby, Engl. Bot. 35: t.2455 f.2. 1813. Fie. 3

Thallus epilithic, continuous, smooth, pale green-brown, rimose. Perithecia

numerous, one-quarter to three-quarters immersed in the thallus, but depth

varies in the same samples, forming low to moderate projections 0.2-0.4 um in

diam. Involucrellum well-developed, hemispherical to conical, dark reddish-

brown. Ascospores ellipsoid-oblong or ellipsoid to narrowly ellipsoid, 15-20 x

6.5-8 um. Conidiomata not observed. Thallus C-, K-, KC-, PD-.

SPECIMENS EXAMINED: TURKEY. GIRESUN: Piraziz, Aydere village, 40°56’33”N

38°06’46’E, 156 m, on siliceous rock, 30 March 2006, leg. K.Kinalioglu, det. A. Orange

(GUB 2389); Trabzon: Arakh, Konak6nii place, sea shore, 40°5717”N 40°02’56’E, 3 m,

on siliceous rock, 12 August 2006, leg. K.Kinalioglu, det. A. Orange (GUB 2390).

ReMARKS— Our Turkish collections had perithecial projections of a similar

size to those described by Orange (2000) and Smith et al. (2009), which

had perithecial projections up to 0.22-0.4 mm. in diam. However, Turkish

ascospores are smaller than those in previous descriptions (15-24 x 6-9.5 um;

Orange 2000, Smith et al. 2009). One Turkish specimen differs ecologically in

occurring on siliceous rocks on the seashore.

Verrucaria elaeina is a common species occurring on a wide variety of

substrata such as shaded sandstone, limestone, calcareous mudstone, concrete,

siliceous rock and brick, in woodland or beneath herbaceous vegetation, on vast

areas of ground in gardens, on damp walls, by unused railways and on ruined

buildings, often on stones embedded in the ground, cliffs, weakly calcareous

rock in shade and occasionally beside streams, also on paths, in the British Isles

and Europe (Smith et al. 2009, Orange 2000). In Turkey, we collected it from

siliceous rocks in moist shady hazelnut gardens and on the seashore. This is the

first report of the species from Asia.

The pale grey-green thallus of Verrucaria elaeina on shaded rocks is

characteristic and often easily recognized by the naked eye. Some poorly

developed morphs have a much less distinctive appearance and could be

New Caloplaca, Hydropunctaria, and Verrucaria spp. for Turkey & Asia... 77

Fic. 3. Verrucaria elaeina (GUB 2389): habit. Scale bar = 1 mm.

confused with V. denudata or V. dolosa. Verrucaria praetermissa differs in its

thicker thallus, more immersed perithecia, slightly larger ascospores, and its

typically freshwater habitat (Smith et al. 2009).

Acknowledgements

I would like to thank Dr. Laszl6 L6k6s and Dr. Michele Piercey-Normore for

critically reviewing the paper, and Dr Alan Orange and Dr. Alexander Khodosovtsev

for the identification of the taxa.

Literature cited

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44(6): 739-741. http://dx.doi.org/10.1017/S0024282912000382

Aslan A, Yazici K. 2013. New Lecanora, Lecidea, Melaspilea, Placynthium, and Verrucaria records

for Turkey and Asia. Mycotaxon 123: 321-326. http://dx.doi.org/10.5248/123.321

Arslan B, Oztiirk, S, Oran S. 2011. Lecanora, Phaeophyscia and Rinodina species new to Turkey.

Mycotaxon 116: 49-52. http://dx.doi.org/10.5248/116.49

78 ... Kinalioglu

Boom PPG van den, Etayo J. 1995. A new epiphytic species of the lichen genus Caloplaca from

southwestern Europe. Mycotaxon 56: 125-132.

Cobanoglu G, Agikgéz B, Baloniu L. 2013. Contributions to lichen diversity of Turkey from the

Sarisu area (Kocaeli). Turkish Journal of Botany 37: 964-969.

http://dx.doi.org/10.3906/bot-1207-23

Goward T, Breuss O, Ryan B, McCune B, Sipman H, Scheidegger C. 1996. Notes on

the lichens and allied fungi of British Columbia. HI. Bryologist 99(4): 439-449.

http://dx.doi.org/10.2307/3244108

Gueidan C, Savi¢ S, Thiis H, Roux C, Keller C, Tibell L, Prieto M, Heidmarsson S, Breuss O,

Orange A, Fréberg L, Wynns AA, Navarro-Rosinés P, Krzewicka B, Pykala J, Grube M, Lutzoni

FE 2009. Generic classification of the Verrucariaceae (Ascomycota) based on molecular and

morphological evidence: recent progress and remaining challenges. Taxon 58(1): 184-208.

Karago6z Y, Aslan A, Yazici K, Aptroot A. 2011. Diplotomma, Lecanora, and Xanthoria lichen species

new to Turkey. Mycotaxon 115: 115-119. http://dx.doi-org/10.5248/115.115

Kanalioglu K, Aptroot A. 2011. Carbonea, Gregorella, Porpidia, Protomicarea, Rinodina,

Solenopsora, and Thelenella lichen species new to Turkey. Mycotaxon 115: 125-129.

http://dx.doi.org/10.5248/115.125

Oran S, Oztiirk $. 2010. Three lichenized fungi new to Turkey. Mycotaxon 112: 389-392.

http://dx.doi.org/10.5248/112.389

Orange A. 2000. Verrucaria elaeina, a misunderstood European lichen. Lichenologist 32: 411-422.

http://dx.doi.org/10.1006/lich.2000.0283

Orange A. 2012. Semi-cryptic marine species of Hydropunctaria (Verrucariaceae,

lichenized Ascomycota) from north-west Europe. Lichenologist 44(3): 299-320.

http://dx.doi.org/10.1017/S00242829 11000867

Rechinger KH. 1951. Phytogeographia Aegaea. Denkschriften, Band. 105, Wien. 208 p.

Senkardesler A, Cansaran DD, Lokés L, Ahti T. 2016. Cladonia trapezuntica (Cladoniaceae,

lichenized Ascomycota): a robust morphotype of Pycnothelia papillaria, a taxonomic

study with conservational survey. Turkish Journal of Botany 40: 104-111.

http://dx.doi.org/10.3906/bot- 1403-49

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA. 2009. The

Lichens of Great Britain and Ireland. British Lichen Society, London. 1046 p.

Vondrak J, Khodosovtsev A, L6kés L, Merkulova O. 2010. The identity of type specimens in BP of

some names in Caloplaca. Mycotaxon 111: 241-250. http://dx.doi.org/10.5248/111.241

Vondrak J, Halici MG, Kocakaya M, Ondrakova OV. 2012. Teloschistaceae (lichenized

Ascomycetes) in Turkey. 1. Some records from Turkey. Nova Hedwigia 94(3-4): 385-396.

http://dx.doi.org/10.1127/0029-5035/2012/0007

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MY COTAXON

January-March 2017— Volume 132, pp. 79-86

http://dx.doi.org/10.5248/132.79

Chloridium terricola sp. nov. from China

YONG WANG"”, CHUN-YU JIE?, KEVIN D. HYDE’,

Yu-LAN JIANG’, TIAN-YU ZHANG & DE-GANG ZHAO*

' Guizhou Key Laboratory Agro-Bioengineering, &

? Department of Plant Pathology, Agriculture College,

Guizhou University, Guiyang, Guizhou 550025, China

> Enshi Tujia and Miao Autonomous Prefecture Academy of Agricultural Sciences,

Enshi, Hubei Province, 445000, China

* Institute of Excellence in Fungal Research and School of Science, Mae Fah Luang University,

Chiang Rai, 57100, Thailand

° Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong 271018, China

* CORRESPONDENCE TO: dgzhao@gzu.edu.cn

ABSTRACT—A new species, Chloridium terricola, was isolated from soil in Sichuan province,

China. Morphologically, it is characterized by the production of pale brown conidia and a

conidiophore bearing a single apical whorl of ampulliform or lageniform phialides. Large

subunit (LSU) rDNA sequence analyses support C. terricola as congeneric with other

Chloridium species.

Key worps—hyphomycetes, ITS, phylogenetic, soil fungi, systematics

Introduction

For competing sexually vs. asexually typified generic names in

Sordariomycetes (except Diaporthales, Hypocreales, and Magnaporthales),

Réblova et al. (2016) have recommended the name Chloridium Link 1809 based

on its priority and widespread use, proposing Gonytrichum Nees & T. Nees

1818 and Melanopsammella Hohn. 1920 as generic synonyms. In 2005, four

hyphomycete isolates morphologically similar to Chloridium were obtained

from soil in the west region of Sichuan province, China. Their taxonomy was

further evaluated through close microscopical examination and large subunit

80 ... Wang & al.

(LSU) rDNA sequence analyses. These morphological and phylogenetic

analyses revealed that the four isolates represented a unique undescribed

Chloridium species, for which we propose the name Chloridium terricola.

Materials & methods

Fungal strains & morphology

Soil samples were collected in Sichuan Province, China, and transported to the

laboratory in sterilized, zip lock polyethylene bags. All isolates are maintained in the

Plant Pathology Herbarium, Guizhou University, Guiyang, China (HGUP). Mycelium

grown from the soil samples was transferred to fresh 1.5% potato dextrose agar (PDA)

medium plates. Cultural characteristics and morphology were determined on PDA

and incubated for 7 d at 25°C under a 12 h/12 h light and darkness cycle, to promote

sporulation. Water solution of 60% (v/v) lactic acid without a color dye was used as the

mounting medium. Dimensions are based on 50 mature conidia and 30 conidiophores

as measured at 100x magnification under a Nikon 90i microscope.

DNA amplification & sequencing

Extracted DNA was amplified via PCR using primers ITS4 and ITS5 for the ITS

region and primers LROR and LRS5 for the LSU region (Vilgalys & Hester, 1990; White

et al. 1990). Additional sequences were obtained from GenBank. After the sequences

were aligned using Clustal X 1.81 (Thompson et al. 1997), the alignments were refined

manually. Our DNA sequences were deposited in GenBank.

Phylogenetic analyses

The sequence data were subjected to parsimony analysis using the heuristic search

option of PAUP” version 4.0b10 (Swofford 2002). MP (maximum parsimony) trees were

inferred using heuristic search option with tree bisection reconnection (TBR) branch

swapping and 1000 random sequence additions with maxtrees at 5000, branches of zero

length collapsed, and all parsimonious trees saved. Measures calculated for parsimony

included tree length (TL), consistency index (CI), retention index (RI), and rescaled

consistence index (RC). Bootstrap analyses (Hillis & Bull 1993) were conducted with

1000 replications.

Taxonomy

Our morphological and molecular phylogenetic analyses support

recognition of our four soil isolates as one phenotypically and phylogenetically

distinct species.

Chloridium terricola Yong Wang bis, Jie & K.D. Hyde, sp. nov. Fie. 1

MycoBank MB 820926

Differs from Chloridium gonytrichii by its single whorl of phialides at the conidiophore

apex and by its light brown conidia.

Type: China, Sichuan Province, Jiuzhaigou, isolated from soil, August 2005, Yu-Lan

Jiang (Holotype, HGUPd4519 [dried culture]; ex-type culture, HGUP4519; GenBank

KM434144, KT893302).

Chloridium terricola sp. nov. (China) ... 81

Fic. 1 (color). Chloridium terricola (HGUPd4519). a: Conidiophores arising from PDA.

b-c: Conidiogenous cells, conidia, and conidiophores. d: Colony on PDA at 7d. e: Conidia.

Scale bars: c-d = 20 um; e = 10 um.

EryMo_oey: Latin, terricola meaning living on the soil.

Cotontgs effuse, velvety, reverse dark brown to black. Mycelium partly

immersed, partly superficial, composed of branched, hyaline, septate, smooth,

3.5-5 um wide hypha. CONIDIOPHORES mononematous, erect arising from

the hypha, hyaline to pale brown, smooth. CONIDIOGENOUS CELLS borne

directly on the hypha or in divergent forks on the conidiophores, ampulliform

82 ... Wang & al.

or lageniform, pale brown, smooth, 7-15 x 2.5-4 um. Conip1A obovoid or

ellipsoid, truncate at the base, pale brown, smooth, 2-3 x 2-2.5 um. Sexual

stage not observed.

ADDITIONAL SPECIMENS EXAMINED: CHINA, SICHUAN PROVINCE: Panzhihua city,

isolated from soil, August 2005, Yu-Lan Jiang (HGUP4509;GenBank KM434143,

KT893301); Chengdu city, isolated from rice soil, August 2005, Yu-Lan Jiang

(HGUP4536; GenBank KM434146, KT893304); Leshan city, isolated from soil, August

2005, Yu-Lan Jiang (HGUP4521; GenBank KM434145, KT893303).

Crytosporiopsis brunnea UAMH 10106 T

Phialocephala glacialis 444 7v T

Phialocephala dimorphospora CBS 300.62 T

Phialocephala botulispora DAOM 75261 T

L Phialocephala lagerbergii CBS 266.33 T

Phialocepahla scopigformis CBS 468.94 T

Phialocepahla sphaeroides UAMH 10279 T

Phialocepahla sp. UAMH 10827

Phialocepahia compacta CBS 507.94 T

Phaeomollisia piceae CdV 2 2.4b T

Acephala applanata CBS 109321 T

Phialocepahia fortinii CBS 443.86 T

96! Phialocepahla fortinii FAP7

Phialocepahla humicola CBS 420.73 T

Phialocepahla xalapensis CBS 218.86 T

Chioridium terricola sp. nov. HGUP4509

Chloridium terricola sp. nov. HGUP4519 T

584% Chioridium terricola sp. nov. HGUP4521

Chioridium terricola sp. nov. HGUP4536

Fic. 2 (above). Topology showing the single most parsimonious tree, inferred from ITS sequence

analysis. Bootstrap values less than 50% are not shown. The tree is rooted with Cryptosporiopsis

brunnea (UAMH 10106). Type specimens are labeled with “T”.

Fic. 3 (right). Topology showing the single most parsimonious tree, inferred from LSU sequence

analysis. Chloridium terricola (in bold) clusters with C. gonytrichii (as Melanopsammella) and

C. viridescens (shown also as Melanopsammella vermicularioides). Bootstrap values less than 50%

are not shown. ‘The tree is rooted with Saccharomyces cerevisiae.

Chloridium terricola sp. nov. (China) ... 83

Saccharomyces cerevisiae Z73326

thdetosphaeria luquillensis AF466074

Striatosphaeria codinaeophora AF466088

50 Brunneodinemasporium brasiliense JQ889288

Pseudolachnea fraxini JQ88930

79 Dendrophoma cytisporoides 1Obt eRe

Codinaeopsis gonytrichoides AF 178556

“*haetosphaeria ovoidea AF064641

Zignoella pulviscula AF466090

4 pnOcHG pulvisciig AF466091

thaetosphaeria ciliata GU180637

—

ire

sa.

97 | Dinemasporium decipiens JQ889291

Dinemasporium hceacbinf ih iV J 08279

92 Wtl| Dinemasporium ‘strigosum JQ88929

Dinemasporium strigosum JQ889299

Dinemasporium strigosum JQ889300

Dinemasporium americana JQ889290

Dinemasporium pseudostrigosum JQ889294

70 | Dinemasporium pseudoindicum JQ889293

Dinemasporium polygonum JQ889292

92 LT Dinemasporium morbidum JQ889296

59 | i Dinemasporium morbidum JQ889297

75 lM Infundibulomyces CUD 113979

ozetella nivea EU825200

Rattania setulifera HM171322

62 Tainosphaeria crassiparies AF466089

87h Melanopsammella ne AF466085

93 IN Melanopsammella vermicularioides AF466086

the Melanopsammella vermicularioides AF466087

95 i Il Chloridium virescens AF064644

100 ill hloridium terricola HGUP4509

| Chloridium terricola HGUP4519

1 i thloridium terricola HGUP 4521

Chloridium terricola HGUP 4536

66 | Melanochaeta hemipsila AY 346292

99 ET! Melanochaeta hemipsila AF466084

100 ee Melanochaeta aotearoae AF466082

iB elanochaeta aotearoae AF466081

100-] | Eilisembia brachypus DQ408563

99 Pyrigemmula aurantiaca HM241692

79 Lécythothecium duriligni AF261071

‘haetosphaeria preussti AF178561

96 lt) Chaetosphaeria myriocarpa AF 466076

*haetosphaeria pygmaea

In Ch ph AF466077

IF ‘haetosphaeria innumera AY017375

- Chloridium lignicola 4

Chaetosphaeria fuegiana EF063574

! Chaetosphaeria callimorpha AF466062

| Chaetosphaeria tropicalis AF466080

rT! Chaetosphaeria sp. AF279418

i Chaetosphaeria ?. AF279419

Cgaetospyaeria fateriphiala AF466070

7 gk ‘haetosphaeria lateriphiala AF466071

‘haetosphaeria lateriphiala fp EeeeT2

‘haetosphaeria hebetiseta

100 Dp AF466

62 haetosphaeria minuta AF466075

Chaetosphaeria cubensis AF466067

g4—___ - Chaetosphaeria conirostris AF466066

65—T Chaetosphaeria decastyla AF466068

66 Chaetosphaeria capitdta AF466061

Wr Chaetosphaeria chlorotunicata AF466064

td Chaetosphaeria biapiculata

Ltt Chaetosphaeria biapiculata AF466065

50 Umbrinosphaeria caesariata AF261069

“*haetosphaeria caesariata AF466060

100 haetosphaeria spinosa AF466079

Ly Chaetosphaeria chalaroides

Ch h halaroides AY 017372

75—CtF | Chaetosphaeria chalaroides AF466063

9 efee ‘haetosphaeria lignomollis AF466073

59 | Chaetosphaeria raciborski AF466078

100 ee Chaetosphaeria sp. AF279416

‘haetosphaeria sp. AF27941

79 Cercophora newfieldiana AF064642

Lasiosphaeria ovina AF064643

84 ... Wang & al.

Phylogenetic analysis

An NCBI-BLAST search of ITS sequences showed a 99% DNA identity

between our isolates and Phialocephala humicola S.C. Jong & E.E. Davis and

P. xalapensis Persiani & Maggi. We generated a phylogenetic tree of Phialocephala

spp. based on ITS sequences (Fic. 2), from which we discovered that our taxon

was close to, but not conspecific with, P humicola and P. xalapensis.

To determine the taxonomic position of our isolates, we used a more

inclusive analysis of the LSU region with Saccharomyces cerevisiae as outgroup

(Crous et al. 2012) and ingroup sequences derived from Crous et al. (2012)

and Fernandez et al. (2006). Of the 1049 characters aligned, 297 are constant,

496 variable characters are parsimony-uninformative, and 256 are parsimony-

informative. One of 32 most parsimonious trees with 1604 steps (CI = 0.610;

HI = 0.390; RI = 0.723) derived from analysis of the LSU sequences is presented

in Frc. 3. Our four Chloridium terricola sequences formed a branch with a

100% BS support (nearly without phylogenetic divergence). They were placed

in Chloridium group with 100% BS, and showed a closer relationship with

Chloridium gonytrichii (F.A. Fernandez & Huhndorf) Réblova & Seifert and

C. virescens (Pers.) W. Gams & Hol.-Jech. with high (95%) BS support.

Discussion

The International Code of Nomenclature for algae, fungi, and plants

(McNeill et al. 2012) eliminated Art. 59, which in previous botanical codes

permitted dual nomenclature for sexual and asexual stages in pleomorphic

fungi. As noted above, Réblova et al. (2016) now accept Melanopsammella as

a synonym of Chloridium. Fernandez & Huhndorf (2005), who treated the

genera before abandonment of Art. 59, noted that Melanopsammella species

are easily distinguished by their small, one-septate ascospores that disarticulate

into partial spores. Sexual stages are almost identical and separation of species

is only possible by noting differences in the asexual stage. Because our isolates

lacked a sexual stage, we base our morphological comparison on asexual

characters.

The obvious morphological difference between Chloridium terricola and the

asexual stage of C. gonytrichii (as Melanopsammella in Fernandez & Huhndorf

2005) is the number of whorls of phialides: C. terricola has only one whorl of

phialides (at the conidiophore apex), whereas C. gonytrichii has one apical whorl

plus 5-8 additional whorls of phialides in the midsection of each conidiophore.

Additionally, conidia are pale brown in C. terricola and light green conidia in

C. (as Melanopsammella) gonytrichii (Fernandez & Huhndorf 2005).

Chloridium terricola sp. nov. (China) ... 85

In comparison to Chloridium terricola, C. virescens (= Melanopsammella

vermicularioides; see Fic. 3) conidia are hyaline, uni-/bi-guttulate and longer

(3.2-4.0 x 2-2.5 um; Gams & Holubova-Jechova 1976).

Phylogenetically, LSU sequence analysis clusters our taxon with (but

taxonomically independent of) C. gonytrichii and C. virescens, supporting

Chloridium terricola as a new taxon.

Acknowledgements

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

by Drs R.F. Castafieda-Ruiz, De-Wei Li, and Shaun Pennycook. We thank Dr Eric

McKenzie for English assistance. This research was supported by the National Science

Foundation of China No. 31560489, Fundamental Research on Science and Technology,

Ministry of Science and Technology of China 2014FY120100, postgraduate education

innovation program of Guizhou Province ZYRC (2014) 004, and the Program for New

Century Excellent Talents in University NCET- 13-0748, the Scientific and Technological

Personnel Training Program of Guizhou Province No. 201341, and Bijie science and

technology project No. (2015) 39.

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Crous PW, Verkley GJ, Christensen M, Castafieda-Ruiz RF, Groenewald JZ. 2012. How important

are conidial appendages? Persoonia 28: 126-137. http://dx.doi.org/10.3767/003158512X652624

Fernandez FA, Huhndorf SM. 2005. New species of Chaetosphaeria, Melanopsammella and

Tainosphaeria gen. nov. from the Americas. Fungal Diversity 18: 15-57.

Fernandez FA, Miller AN, Huhndorf SM, Lutzoni FM, Zoller S. 2006. Systematics of the genus

Chaetosphaeria and its allied genera: morphological and phylogenetic diversity in north

temperate and neotropical taxa. Mycologia 98: 121-130.

http://dx.doi.org/10.3852/mycologia.98.1.121

Gams W, Holubova-Jechova V. 1976. Chloridium and some other dematiaceous hyphomycetes

growing on decaying wood. Studies in Mycology 13:1-99.

Hillis DM, Bull JJ. 1993. An empirical test of bootstrapping as a method for assessing confidence

in phylogenetic analysis. Systematic Biology 42: 182-192.

http://dx.doi.org/10.1093/sysbio/42.2.182

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. 2012. International Code of Nomenclature

for algae, fungi, and plants. Regnum vegetabile 154.

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MA, Cannon PF, Daranagama DA, De Beer ZW, Huang SK, Hyde KD, Jayawardena R,

Jaklitsch W, Jones EBG, Ju YM, Judith C, Maharachchikumbura SSN, Pang KL, Petrini

LE, Raja HA, Romero AI, Shearer C, Senanayake IC, Voglmayr H, Weir BS, Wijayawarden

NN. 2016. Recommendations for competing sexual-asexually typified generic names in

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131-153. http://dx.doi.org/10.5598/imafungus.2016.07.01.08

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

4.0b10. Sunderland, Massachusetts: Sinauer Associates.

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

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

Nucleic Acids Research 24: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876

Vilgalys R, Heste 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 S, Taylor J, 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR

protocols: a guide to methods and applications. New York: Academic Press, U.S.A.

http://dx.doi.org/10.1016/B978-0-12-372180-8.50042-1

MY COTAXON

January-March 2017—Volume 132, pp. 87-94

http://dx.doi.org/10.5248/132.87

Erysiphe russellii in Korea supported

by new morphological and sequence analyses

Tur THUONG THUONG NGUYEN & HYANG BURM LEE*

College of Agriculture & Life Sciences, Chonnam National University,

Gwangju 61186, Republic of Korea

*CORRESPONDENCE TO: hblee@jnu.ac.kr

AsBstTRACT—Morphological and rDNA ITS sequence analyses were conducted on three

strains obtained from leaf lesions on creeping wood sorrel (Oxalis corniculata) covered

by a powdery mildew. No chasmothecia were observed, but the symptoms and traits of

conidiophores and conidia agree well with those of Erysiphe russellii (previously recorded

from Korea as Microsphaera russellii). Phylogenetic analysis shows that the three strains

share 97.9-98.06 % identity with Erysiphe pisi (FJ378867) but form a separate E. russellii clade

with a sequence of that species based on Japanese material. This supports the occurrence of

Erysiphe russellii in Korea.

Key worps—Erysiphales, pathogen, phylogeny, taxonomy

Introduction

Species in the genus Erysiphe (Ascomycota, Erysiphales, Erysiphaceae)

are plant pathogens that cause powdery mildews (Takamatsu et al. 2015).

The Erysiphaceae currently comprises about 873 species belonging to 16

genera (Braun & Cook 2012). It includes five tribes and two basal genera,

which have been described by Mori et al. (2000) and Takamatsu et al. (2005).

Creeping wood sorrel (Oxalis corniculata, Oxalidaceae) is a generally low-

growing, spreading, perennial weedy or invasive plant with a yellow flower

and rounded trifoliate clover-like leaves. It is widely distributed worldwide

and occurs in diverse habitats, including arable fields and gardens or

wastelands, but its origin is unknown. The leaves of the creeping wood sorrel

are sometimes used as edible plant; it has a tangy taste due to a rich amount

88 ... Nguyen & Lee

TABLE 1. Erysiphe samples analyzed.

SPECIES

E. diffusa

E. glycines

E. howeana

E. palczewskii

E. peruviana

E. pisi

E. polygoni

E. sp. [1]

E. sp. [2]

E. trifoliorum

NI: Not identified.

SAMPLE NO.

SMK15414

MUMH1162

MUMH791

MUMH1463

MUMH1451

MUMH56

MUMH52

PMWI

PMQB

DAG08-36

08-3618S

FF 06

GH 07

GH 04

Lathyrus 07

GE 07

bean 2A

DNA14

MUMH2431

UC1512295

UC1512308

EB 2004

DB 121211

EML-COR2

EML-COR3

MUMH0105

VPRI 22166

MUMH29s

SOURCE (HOST)

Glycine soja

Glycine max

Amphicarpaea edgeworthii

var. japonica

Desmodium oxyphyllum

Oenothera biennis

Caragana arborescens

Tecoma capensis

Pea

Lathyrus latifolius

Phaseolus vulgaris

Rumex obtusifolius

Polygonum sp.

Polygonum arenastrum

Rumex crispus

Phaseolus vulgaris

Oenothera biennis

Senna septentrionalis

Oxalis corniculata

Trifolium repens

Trifolium vulgaris

Two unknown species are designated by [1] (still unknown) and

[2] (supported as E. russellii after complete analysis).

LOCALITY

South Korea

Japan

USA

Vietnam

USA

Mexico

Japan

Argentina

USA

Brazil

Italy

Mexico

Korea

Japan

China

GENBANK ITS No.

ABO78812

ABO078813

ABO078800

ABO078810

ABO078809

ABO015934

ABO015927

AFO11301

GQ497277

GQ497276

GU117107

GU987124

FJ378867

FJ378872

FJ378870

FJ378879

FJ378869

AF011306

GU570595

LC009892

LC010013

AF011307

AFO011308

AY739109

JQ288740

JQ730709

KP941751

KP941752

LC009922

JQ282732

AF298542

ABO015913

Erysiphe russellii molecularly supported in Korea ... 89

of vitamin C and is traditionally used for drinks by infusing the leaves in hot

water, sweetening, and then chilling.

Between July and October 2010 and in August 2012, symptoms of powdery

mildew were observed on creeping wood sorrel plants in Suwon, Gwangju

and Cheongyang in Korea. Erysiphe russellii (Clinton) U. Braun & S. Takam.

has been reported from several species of Oxalis worldwide (Farr & Rossman

2015) and is the only powdery mildew that has been previously reported on

creeping wood sorrel in Korea, based on morphology of the asexual morph.

Therefore, the morphology of the new Korean powdery mildew collections

on Oxalis corniculata has been re-examined, supplemented by a molecular

phylogenetic analysis of rDNA ITS sequences, to confirm the identity of the

causal agent of the powdery mildew as E. russellii. Molecular analyses of the

internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) is a well-

established method for fungal identifications at species level (White et al.

1990), which have been frequently applied to powdery mildews (Takamatsu

et al. 1999, Cunnington et al. 2003).

Materials & methods

Sampling and morphological observations

Fresh samples infected by the powdery mildew pathogen were collected and

examined by stereo and light microscopy. Voucher specimens were deposited in

EML (Environmental Microbiology Laboratory) Fungarium (EMLF; Chonnam

National University, Gwangju, Korea as EML-COR1 (obtained from a garden

located in Suwon in July 2010), EML-COR2 (obtained from a pot for red pepper in

Gwangju in August 2010), and EML-COR3 (obtained from a greenhouse located in

Cheongyang in late October 2010).

DNA extraction, polymerase chain reaction amplification, and sequencing

Total genomic DNA was isolated from 50-100 mg of powdery mildew conidia

and mycelia using the Accuprep® Genomic DNA Extraction Kit (Bioneer Corp.,

Daejeon, Korea). The rDNA ITS region was amplified using primers ITS1F and LR5F

following Lee (2012a). The 20 uL PCR amplification mixture contained 2 uL DNA

template, 1.5 uL per primer (5 pM/uL), 1 uL Accupower® PCR premix (containing

Taq DNA polymerase, dNTPs, buffer, and tracking dye; Bioneer Corp.), and 14 uL

of sterile water. Amplification parameters were an initial denaturation step at 95°C

for 5 min, 30 cycles of denaturation at 94°C for 1 min + annealing at 54°C for 30 s

+ extension at 72°C for 1 min, and a final extension step at 72°C for 10 min. The

PCR products were purified using the Accuprep® PCR Purification Kit (Bioneer

Corp.). DNA sequencing was performed in an ABI 3700 Automated DNA sequencer

(Applied Biosystems Inc.).

90 ... Nguyen & Lee

Phylogenetic analyses

After initial sequence alignment using CLUSTAL_X (Thompson et al. 1997), the

alignment was edited manually. Sequence analyses were performed using MEGA 6

(Tamura et al. 2013) with the default settings. Phylogenetic trees were generated from

the data using maximum-likelihood (ML), neighbor joining (NJ), and maximum

parsimony (MP) methods. The Kimura 2-parameter model was selected as the best

model to construct trees for NJ. Nearest-neighbor- interchange (NNI) was selected

for the ML heuristic method, and the initial tree for ML was set automatically.

The MP analysis was conducted with the heuristic search option using the tree-

bisection-reconstruction (TBR) algorithm. All sites were treated as unordered and

unweighted, with gaps treated as missing data. The strength of the internal branches

of the resulting tree was tested with a bootstrap (BS) analysis using 1000 replications

in the ML, NJ, and MP analyses. Tree scores, including tree length, consistency index

(CI), and retention index (RI), were also calculated. Erysiphe glycines was used as

outgroup. Sequences from this study and those from GenBank database are listed

in TABLE 1.

Taxonomy

Erysiphe russellii (Clinton) U. Braun & S. Takam., Schlechtendalia 4: 13 (2000) Fie. 1

= Microsphaera russellii Clinton, in Peck, Rep. (Annual)

New York Stat. Mus. 26: 80 (1874).

= Oidium oxalidis McAlpine, Proc. Roy. Soc. Victoria, N.S., 6: 219 (1894).

Mycelium on leaves, effuse, covering the entire leaf surface, white, thin,

powdery. Hyphae somewhat straight to slightly wavy, septate, branched.

Appressoria lobed and single or in opposite pairs. Conidiophores on top

of hyphal mother cells, erect, 76.6-134 x 4.7-6.8 um, with lecythiform to

clavate apices above two or three cells. Conidia formed singly; primary

conidia ellipsoidal to fusiform; secondary conidia, slender, cylindrical

to oblong with slightly obtuse or truncate apices, 24-55 x 13.5-22 um.

Chasmothecia not observed.

SPECIMENS EXAMINED—KOREA, Cheongyang, 36°27°45”"N 126°51’34”E (EML-COR3),

Gwangju, 35°10°0”N 126°55’°0"E (EML-COR2), Suwon, 37°17'27"N 127°00°32”E

(EML-COR1), on Oxalis corniculata L., October 2010, H.B. Lee. Voucher specimens

(KOSPFG0000134896) were deposited in the herbarium of the National Institute of

Biological Resources (NIBR), Incheon, Korea.

Discussion

In Korea, Erysiphe russellii was previously reported by Shin (2000,

as Microsphaera russellii based on conidial morphology) as the causal

pathogen on creeping wood sorrel. Previous studies (Takamatsu et al.

2003, 2015; Lee 2012a,b, 2013a,b, 2015; Lee et al. 2013) have successfully

Erysiphe russellii molecularly supported in Korea... 91

Fic. 1 Powdery mildew symptoms and the morphology of the causal pathogen, Erysiphe sp.

EML-COR3 isolate. A. Powdery mildew symptoms on the leaves of creeping wood sorrel;

B & C. Conidia and conidia with germ tube (yellow arrow); D. Appressoria on hyphae (yellow

arrow). E-G. Conidiophores. Bars: B, D, F, G =20 um; C, E=50 um.

used rDNA ITS sequence data to identify Erysiphe species. Therefore,

we conducted phylogenetic analysis based on three Korean strains and a

single strain from Japan (LC009922) to confirm the identity of the Korean

collections. ITS sequences from the three Korean strains were compared to

those from related species retrieved from GenBank. Our sequence analysis

(Fic. 2) clustered the sequences from strains EML-COR1, EML-COR2, and

EML-COR3 together with a Japanese sequence in a separate clade in the

phylogenetic tree assignable to Erysiphe russellii, supporting it as a distinct

species. The morphology of the asexual morph most closely resembled that

of E. russellii described by Braun (1987) and Shin (2000).

In the phylogenetic analyses, two small clades, highlighted by question

marks emerged which could not be satisfactorily named (Fie. 2).

Various additional powdery mildew species have been reported on

species of Oxalis worldwide, e.g. Golovinomyces orontii (Castagne) Heluta

(= Erysiphe polyphaga Hammarl.), Leveillula oxalidicola T.Z. Liu & U. Braun,

and a species of Phyllactinia (Braun & Cook 2012, Farr & Rossman 2015).

This report is the first to verify the presence of E. russellii on creeping wood

sorrel in Korea by DNA sequence comparisons. The pathogenicity of the

biotrophic fungus was confirmed through inoculation experiments made by

gently pressing infected leaves onto mature leaves of healthy creeping wood

sorrel plants in a glasshouse. Seven days after inoculation, symptoms similar

to those observed under natural infection were developed on the inoculated

leaves of creeping wood sorrel plants (data not shown). More studies on

92 ... Nguyen & Lee

Erysiphe diffusa ABO78812

Erysiphe diffusa ABO78810

Erysiphe diffusa ABO78809

Erysiphe diffusa ABO78800

E. diffusa

Erysiphe diffusa ABO78813

7 Erysiphe sp. EB2004 AY739109

Erysiphe sp. 3D JQ730709 ?

Erysiphe polygoni GU570595

Erysiphe peruviana GU117107 }

100 ' Erysiphe peruviana GU987124

Erysiphe howeana AFQ11301

99! Erysiphe sp. DB121211 JQ288740 | E. howeana

Erysiphe pisi FJ378870

Erysiphe pisi FJ378867

Erysiphe pisi AFO11306

79) Erysiphe pisi FJ378879

Erysiphe pisi FJ378872

Erysiphe pisi FJ378869

Erysiphe sp. MUMH0105 LC009922

Erysiphe sp. EML-COR3 KP941752

99| Erysiphe sp. EML-COR1 KP941750

Erysiphe sp. EML-COR2 KP941751

98 | Erysiphe palczewskii GQ497277 |

Erysiphe palczewskii GQ497276

Erysiphe trifoliorum ABO15913

Erysiphe trifolii JQ282732 |

95" Erysiphe trifolii AF298542

87 | Erysiphe polygoni LC010013

Erysiphe polygoni AF011307

a7 Erysiphe polygoni AF011308

97" Erysiphe polygoni LC009892

Erysiphe glycines AB015927

100 Erysiphe glycines ABO15934

E. pisi

E. russellii

E. palezewshkii

E. trifoliorum

E. polygoni

Fic. 2 Phylogenetic status of the EML-COR1, EML-COR2, and EML-COR3 and MUMH0105

isolates and related species based on maximum parsimony (MP) of the internal transcribed

spacer (ITS) region sequences. Numbers at the nodes indicate the bootstrap values (>50 %)

from 1000 replications. Erysiphe glycines was selected as outgroup. The thick line is based on

a previous phylogenetic system constructed by Takamatsu et al. (2015). Two small clades,

highlighted by question marks, emerged that could not be satisfactorily named. The scale bar

indicates the number of nucleotide substitutions per site.

the distribution of powdery mildews on Oxalis in Korea, including a search

for the sexual morph, are necessary. Moreover, sequence data based on

North America samples (USA is the type locality of E. russellii) and material

from Australia (type locality of Oidium oxalidis) are urgently required for

comparison.

Erysiphe russellii molecularly supported in Korea ... 93

Acknowledgments

This work was supported by a grant from the National Institute of Biological

Resources (NIBR), funded by the Ministry of Environment (MOE), Republic of

Korea. Authors are grateful to Dr. Paul Kirk (Royal Botanic Garden at Kew, U.K.)

and Dr. Jeffrey Stone (Oregon State University, Corvallis, USA) for kind review of the

paper and to Prof. Dr Susumu Takamatsu (Mie University, Japan) for kind supply of

sequence data of a Japanese strain.

Literature cited

Braun U. 1987. A monograph of the Erysiphales (powdery mildews). Beiheft zur Nova Hedwigia

89: 1-700.

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildews). CBS

Biodiversity Series 11: 707 p.

Cunnington JH, Takamatsu S, Lawrie AC, Pascoe IG. 2003. Molecular identification of

anamorphic powdery mildews (Erysiphales). Australasian Plant Pathology 32: 421-428.

http://dx.doi.org/10.1071/AP03045

Farr DF, Rossman AY. 2015. Fungal Databases, Systematic Mycology and Microbiology

Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/

Lee HB. 2012a. First report of powdery mildew caused by Erysiphe arcuata on lanceleaf

coreopsis (Coreopsis lanceolata) in Korea. Plant Disease 96: 1827 p.

http://dx.doi.org/10.1094/PDIS-08-12-0754-PDN

Lee HB. 2012b. Molecular phylogenetic status of Korean strain of Podosphaera xanthii, a causal

pathogen of powdery mildew on Japanese thistle (Cirsium japonicum) in Korea. Journal of

Microbiol 6: 1075-1080. http://dx.doi.org/10.1007/s12275-012-2618-z

Lee HB. 2013a. First report of Oidium anamorph of Erysiphe hypophylla causing powdery

mildew on leafy lespedeza (Lespedeza cyrtobotrya) in Korea. Plant Disease 97: 287 p.

http://dx.doi.org/10.1094/PDIS-08-12-0774-PDN

Lee HB. 2013b. First report of powdery mildew caused by Erysiphe heraclei on curled dock

(Rumex crispus) in South Korea. Plant Disease 97: 427 p.

http://dx.doi.org/10.1094/PDIS-10-12-0904-PDN

Lee HB. 2015. First report of powdery mildew caused by Erysiphe alphitoides on Quercus acutissima

on Korea. Plant Disease 99: 889 p. http://dx.doi.org/10.1094/PDIS-08-14-0848-PDN

Lee HB, Kim CJ, Mun HY. 2013. First report of powdery mildew on Spanish needles

(Bidens bipinnata) caused by Podosphaera xanthii in Korea. Plant Disease 97: 1385 p.

http://dx.doi.org/10.1094/PDIS-10-12-0966-PDN

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

Shin HD. 2000. Erysiphaceae of Korea. Plant pathogens of Korea 1. NIAST. Suwon. Korea.

Tamura K, Stecher G, Peterson D, Filioski A, Kumar S. 2013. MEGA6: Molecular

evolutionary genetics analysis version 6.0. Molecular Biology Evolution 30: 2725-2729.

http://dx.doi.org/10.1093/molbev/mst197

Takamatsu S, Tetsuya H, Yukio S, Yukihiko N. 1999. Phylogenetic relationships of Microsphaera

and Erysiphe section Erysiphe (powdery mildews) inferred from the rDNA ITS sequences.

Mycoscience 40: 259-268. http://dx.doi.org/10.1007/BF02463963

94 ... Nguyen & Lee

Takamatsu S, Sato Y, Mimuro SK. 2003. Erysiphe wadae: a new species of Erysiphe sect. Uncinula

on Japanese beech. Mycoscience 44: 165-171. http://dx.doi.org/10.1007/s10267003-0100-9

Takamatsu S, Braun U, Limkaisang S. 2005. Phylogenetic relationships and generic affinity

of Uncinula septata inferred from nuclear rDNA sequences. Mycoscience 46: 9-16.

http://dx.doi.org/10.1007/s10267-004-0205-9

Takamatsu S, Ito H, Shiroya Y, Kiss L, Heluta V. 2015. First comprehensive phylogenetic analysis

of the genus Erysiphe (Erysiphales, Erysiphaceae) 1. The microsphaera lineage. Mycologia

107: 475-489. http://dx.doi.org/10.3852/15-007

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

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

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

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

ribosomal RNA gene for phylogenetics. 315-322 in Innis MA, Gelfand DH, Sninsky JJ,

White TJ (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

MY COTAXON

January-March 2017— Volume 132, pp. 95-97

http://dx.doi.org/10.5248/132.95

Phlebia brevibasidia sp. nov. from India

GURPREET KAUR, AVNEET P. SINGH * & G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT—A new agaricomycetous species, Phlebia brevibasidia, is described from Punjab,

India.

Key worps—Meruliaceae, tropical, Anandpur Sahib, Phlebia cremeoalutacea, Phlebia

crassisubiculata

During September 2015, Gurpreet and Avneet collected an unknown

agaricomycetous fungus associated with an angiospermous log from Dabri in

district Anandpur Sahib, Punjab, India. After comparison of macroscopic and

microscopic characters (Rattan 1977; Eriksson et al. 1978, 1981; Bernicchia

& Gorjon 2010; MycoBank 2016; Sharma 2012; Singh et al. 2010), it has

been identified as a tropical species differentiated from temperate Phlebia

cremeoalutacea and P. crassisubiculata by conspicuous morphological

differences. A portion of the basidiocarp was sent to Dr. Nils Hallenberg

(Denmark), who confirmed the findings. The holotype has been conserved in

the Herbarium, Botany Department, Punjabi University, Patiala, India (PUN).

Phlebia brevibasidia G. Kaur, Avn.P. Singh & Dhingra, sp. nov. PratEs 1, 2

MycoBank MB 816998

Differs from Phlebia cremeoalutacea by its bigger basidiospores, its stalked short basidia,

its prominently projecting encrusted cystidia, and its compact pseudoparenchymatous

context; and from P. crassisubiculata by its stalked short basidia and its thin subiculum.

Type: India, Punjab, Anandpur Sahib, Dabri, on angiospermous log, 30 July 2015,

Gurpreet & Avneet 7932 (PUN, holotype).

Erymo.oey: The epithet refers to short basidia

Basidiocarp resupinate, adnate, effused, <230 um thick in section, hymenial

surface smooth, orange white to pale orange to grayish orange when fresh,

not changing much on drying; margins thinning, fibrillose, paler than the

96 ... Kaur, Singh & Dhingra

PiaTE 1. Phlebia brevibasidia (holotype, PUN). 1. Basidiocarp showing hymenial surface.

color of the hymenial surface or indeterminate. Hyphal system monomitic.

Generative hyphae clamped; subiculum <40 um thick, of thick-walled,

less branched, horizontal, <4 um wide hyphae; context and subhymenium

pseudoparenchymatous, <190 um thick, of thin-walled, vertical, more

branched, densely packed hyphae, <3.5 um wide. Cystidia 58-101 x 12.8-17.5

um, prominent, subfusiform, thick-walled, with basal clamp, encrusted with

crystalline matter; projecting up to 50 um out of hymenium. Basidia 15-18

x 5-7.3 um, clavate, stalked, slightly thick-walled, 4-sterigmate, with basal

clamp; sterigmata <3.8 um long. Basidiospores 4.5-7.3 x 3.5-4.5 um, ellipsoid,

smooth, inamyloid, acyanophilous.

REMARKS—Phlebia cremeoalutacea (Parmasto) K.H. Larss. & Hjortstam, a

temperate species, differs in having subclavate-clavate basidia (>20 um long),

cystidia either enclosed and non-encrusted or projecting and encrusted, and

smaller basidiospores (3-4.5 x 2-2.5 um; Eriksson et al. 1981). Another temperate

species, P. crassisubiculata Avn.P. Singh et al., differs in having longer, clavate

basidia and a distinctly thicker subiculum (<120 um thick; Singh et al. 2010).

Acknowledgements

The authors thank SERB, DST, Government of India for financial assistance, Head,

Department of Botany, Punjabi University, Patiala for providing research facilities, Dr.

Nils Hallenberg (Sweden) for expert comments and peer review and Prof. B.M. Sharma

(Department of Plant Pathology, COA, CSKHPAU, Palampur, H.P., India) for peer

review and Mycotaxon Ltd., for underwriting publication charges.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.]. Fungi Europaei 12. Edizioni Candusso. Alassio.

Italia. 1008 p.

ly Da

Mb Pate

Phlebia brevibasidia sp. nov. (India) ... 97

a ay, )

arate: ig

a7? TAL 7, WP Q F an

Waar Vk i TT NEN TY,

/ WA ah Nit Neer ANG

\ DY AVIRONT 2:

S \ 4 C iS BANS <

SOME NaNINAN BPOKR) |S

VAN NNN AN WN

YANNIS oD

ST QA IRV SBE

/ BR

Y ONS 2d TRESS

PLaTE 2. Phlebia brevibasidia (holotype, PUN).

1. Basidiospores 2. Vertical section through basidiocarp.

Eriksson J, Hjortstam K, Ryvarden L. 1981. The Corticiaceae of North Europe. Vol. 6. Phlebia -

Sarcodontia. Fungiflora, Oslo. pp. 1049-1276.

MycoBank. 2016. Fungal databases. Nomenclature and species banks. [Accessed: 26/03/2015].

Rattan SS. 1977. The resupinate aphyllophorales of the North Western Himalayas. Bibliotheca

Mycologica 60. 427 p.

Sharma JR. 2012. Aphyllophorales of Himalaya. Botanical Survey of India, Calcutta. 590 p.

Singh AP, Priyanka, Dhingra GS, Singla N. 2010. A new species of Phlebia (Basidiomycetes) from

India. Mycotaxon 112: 21-24. https://doi.org/10.5248/112.21

MY COTAXON

January-March 2017—Volume 132, pp. 99-106

http://dx.doi.org/10.5248/132.99

Two new records of puffballs in Thailand

JATURONG KUMLA, NAKARIN SUWANNARACH & SAISAMORN LUMYONG*

Department of Biology, Faculty of Science, Chiang Mai University,

Chiang Mai, 50200, Thailand

* CORRESPONDENCE TO: saisamorn.l@cmu.ac.th

AsBstRACT—Calvatia holothurioides and Morganella puiggarii, two puffballs in the

Agaricaceae, are reported for the first time from Thailand. The specimens were identified

based on morphological and ITS sequence analyses. Descriptions and illustrations are

provided.

Key worps—Agaricales, basidiomycete, phylogeny, taxonomy

Introduction

Puffballs in the genera Bovista, Bovistella, Calvatia, Lycoperdon, Morganella,

Tulostoma, Utraria, and Vascellum, formerly classified as Lycoperdaceae, are

now included in Agaricaceae (Kirk et al. 2008). Recent molecular phylogenetic

studies have proposed a widening of Lycoperdon to incorporate traditional

genera such as Bovistella, Morganella, Utraria, and Vascellum (Larsson &

Jeppson 2008, Gube 2009). During a taxonomic survey of puffballs in northern

Thailand, we collected specimens that corresponded to Calvatia holothurioides

and Morganella puiggarii, species previously unknown in the country. Here we

present the morphological characters and ITS sequence analyses of the Thai

specimens.

Materials & methods

Morphology

Basidiomes were collected from Doi Suthep-Pui and Doi Inthanon National

Parks, Chiang Mai Province, Thailand in 2013, and wrapped in aluminum foil or

100 ... Kumla, Suwannarach & Lumyong

placed in plastic specimen boxes for transport to the laboratory, where notes on

macromorphological features and photographs were taken within 24 h. Color names

and codes follow Kornerup & Wanscher (1978). The specimens were dried at 40—45

°C and deposited in the herbarium of the Research Laboratory for Excellence in

Sustainable Development of Biological Resources, Faculty of Science, Chiang Mai

University, Thailand (SDBR-CMU). Tissues from dried specimens were rehydrated

in 95% ethanol followed by distilled water, 3% KOH, or Melzer’s reagent for

microscopical examination. Dimensions are based on at least 50 measurements of

each anatomical structure. Scanning electron images of basidiospores were captured

with a JEOL JSM-5910LV electron microscope.

DNA extraction & amplification

DNA was extracted from fresh specimens using a Favorgen’ Genomic DNA

Extraction Mini Kit. The internal transcribed spacer (ITS) region of ribosomal RNA

gene was amplified by the polymerase chain reaction (PCR) with ITS4 and ITS5

primers under the following thermal conditions: 94 °C for 2 min; 35 cycles of 95 °C

for 30 s, 50 °C for 30 s, 72 °C for 1 min; and finally 72 °C for 10 min. PCR products

were checked on 1% agarose gels stained with ethidium bromide under UV light and

purified using NucleoSpin’ Gel and PCR Clean-up Kit following the manufacturer's

protocol. The purified PCR products were directly sequenced. Sanger sequencing

was performed by 1°’ Base Company (Kembangan, Malaysia) using the same ITS4

and ITS5 PCR primers. The new sequences were used to query GenBank database

via BLAST (http://blast.ddbj.nig.ac.jp/top-e.html).

Phylogenetic analysis

A multiple alignment subroutine in Clustal X (Thompson et al. 1997) and

phylogenetic tree was generated using the PAUP beta 10 vers. 4.0 (Swofford 2002).

In maximum parsimony analysis, all characters were equally weighted and gaps

were treated as missing data. Heuristic searches were performed with 100 random-

addition sequence replicates and tree-bisection-reconnection branch swapping.

Bootstrap analysis (Felsenstein 1985) was conducted with 1000 replicates using the

same settings as above.

Taxonomy

Calvatia holothurioides Rebriev, Mikol. Fitopatol. 47: 21 (2013). FIG. 1

Basidiomes pyriform, turbinate to broadly excipuliform, 3-6 cm diam and

4-6 cm high. Exoperidium two-layered, brownish orange (6C6) to brown

(7E4), slightly wrinkled, formed by hyaline, septate and branched hyphae,

4—7 um wide, intermixed with hyaline, thin-walled vesicles 12-20 um diam,

with an underlying pseudoparenchymatous layer. Endoperidium light

brown (7D6), formed by septate, thin-walled hyphae, 3-5 um diam. Gleba

cottony, yellowish white (4A2) when young, light brown (7D6) when mature.

Subgleba light brown (7D6), composed of compacted cells. Paracapillitium

Calvatia & Morganella spp. new for Thailand ... 101

Fic. 1. Calvatia holothurioides (SDBR-CMU-NK0O152). A: basidiomes; B: exoperidial layer;

C: paracapillitial hyphae; D: basidiospores (LM); E: basidiospore (SEM). Scale bars: A = 10 mm,

B=10um, C=5 um, D=2 um, E=1 um.

thin-walled, septate, branched, hyaline, 3-5 um diam. Pores roundish, <1.5

um diam. Basidiospores ellipsoid to elongate, 3-5.5 x 2-3 um (without

ornamentation), Q = 1.5-1.8; hyaline in water and KOH, unchanging in

Melzer’s reagent; echinate, shortly pedicellate. Spines 0.2-0.5 um long.

SPECIMEN EXAMINED—THAILAND, CHIANG MAI PROvINCcE, Muang District,

Doi Suthep-Pui National Park, 18°47’34”N 98°55’49’E, elevation 746 m, on soil in

deciduous forest, 1 June 2013, Suwannarach N. & Kumla J. (SDBR-CMU-NKO0152,

GenBank KX064242).

Morganella puiggarii (Speg.) Kreisel & Dring, Feddes Repert 74: 116 (1967). Fic. 2

Basidiomes globose to subglobose, 0.6-—2 cm diam. and 0.9-2.3 cm high,

with whitish rhizomorphs attached at the base. Exoperidium white (3A1) to

yellowish white (32) when fresh, dark brown (6F6) when dried, covered by

minute conical tubercles, composed of chains of globose cells, 8-22 x 12-28

um diam. Endoperidium soft, white (3A1). Gleba when young cottony and

white (3A1), when mature powdery and dark brown (7F7). Pseudocolumella

present. Subgleba orange-white (5A2), composed of compacted cells.

102 ... Kumla, Suwannarach & Lumyong

Fic. 2. Morganella puiggarii (SDBR-CMU-NKO141). A, B: basidiomes; C: exoperidial elements;

D: paracapillitial hyphae; E: basidiospores (LM); F: basidiospores (SEM). Scale bars: A, B= 10mm,

C= 10um, D=5 um, E=5 um, F=5 um.

Paracapillitium thin-walled, 1-2.5 um diam., septate, branched, hyaline,

covered by an amorphous hyaline incrustation. Basidiospores globose, 3-4

um diam. (without ornamentation), Q = 1-1.05; pale orange (5A3) in water

and KOH, unchanging in Melzer’s reagent; echinate, shortly pedicellate.

Spines <0.5 um long.

SPECIMEN EXAMINED— THAILAND, CHIANG MAI PROovINcE, Chom Thong District,

Doi Inthanon National Park, 18°34’16”N 98°28’56’E, elevation 235 m, on rotting wood

in mixed evergreen hill forest, 12 July 2013, Suwannarach N., Kumla J. & Lumyong S.

(SDBR-CMU-NK0152, GenBank KX064241).

ITS sequence and phylogenetic analysis

The ITS sequences of C. holothurioides SDBR-CMU-NK0152 (659 bp) and

M. puiggarii SDBR-CMU-NK0141 (699 bp) were deposited in GenBank. The

Calvatia & Morganella spp. new for Thailand ... 103

similarity search revealed a 98% similarity between SDBR-CMU-NKO152

and C. holothurioides JQ734547 and KJ909662 and a 96% similarity between

SDBR-CMU-NKO141 and M. purpurascens (Berk. & M.A. Curtis) Kreisel &

Dring KC414581.

In the phylogenetic analysis, the aligned dataset of 30 ITS sequences

comprised 745 characters, of which 454 characters were constant, 76 variable

characters were parsimony uninformative, and 215 characters were parsimony

100) 71 X064241 Morganella puiggarii

84 KC414581 Morganella purpurascens

76 AJ237626 Morganella subincarnata

AF485065 Morganella fuliginea

DQ112566 Lycoperdon molle

DQ112583 Lycoperdon decipiens

83--DQ112576 Lycoperdon lambinonii

DQ112560 Lycoperdon frigidum

DQ112632 Lycoperdon marginatum

93 DQ112630 Lycoperdon perlatum

65 DQ112631 Lycoperdon norvegicum

199/2Q112554 Vascellum pratense

991 'DQ112556 Vascellum intermedium

HQ235048 Vascellum curtisii

1997 PQ112558 Morganella pyriformis

DQ112557 Morganella pyriformis

KX064242 Calvatia holothurioides

100|KJ909662 Calvatia holothurioides

39 JQ734547 Calvatia holothurioides

AF485064 Calvatia rubroflava

DQ112625 Calvatia craniiformis

DQ112624 Calvatia candida

ne AJ486962 Calvatia fragilis

61 AJ486868 Calvatia cyathiformis

ef —AJ237613 Bovista polymorpha

88 AJ237631 Bovista pusilla

75\'|_1Q112613 Bovista plumbea

DQ112610 Bovista cretacea

DQ415732 Tulostoma squamosum

DQ112629 Tulostoma kotlabae

Lycoperdon sensu lato

100

Calvatia

Bovista

Fic. 3. One of 62 maximum parsimonious trees of 30 ITS sequences. Tulostoma kotlabae and

T. squamosum were used as the outgroup. Numbers above branches show bootstrap supports

>50%; scale bar represents ten substitutions per nucleotide position. The sequences obtained

in this study are in bold.

104 ... Kumla, Suwannarach & Lumyong

informative. Heuristic searches produced a length of 667 steps, CI = 0.609,

RI = 0.686, RC = 0.417 and HI = 0.391. One of 62 maximum parsimonious trees

(Fic. 3) clustered our C. holothurioides ITS sequence with C. holothurioides

sequences from GenBank within the Calvatia clade. Our sequence of Morganella

puiggarii was assigned to the Lycoperdon sensu lato clade and formed a sister

taxon to M. purpurascens with 100% bootstrap support.

Discussion

Calonge et al. (2003) assigned C. holothurioides to “Calvatia sect.

Sporocristata” based on its echinulate and ellipsoidal spores. This invalid

section contained only two species, C. oblongispora V.L. Suarez et al. and

C. sporocristata Calonge (Calonge et al. 2003, Suarez et al. 2009). Calvatia

holothurioides clearly differs from C. oblongispora and C. sporocristata by its

pyriform to broadly excipuliform basidiomes and smaller spores (Calonge et

al. 2003; Suarez et al. 2009; Rebriev 2013). Calvatia holothurioides has been

reported previously from Vietnam (Rebriev 2013).

Morganella puiggarii differs from M. arenicola Alfredo & Baseia,

M. albostipitata Baseia & Alfredo, M. benjaminii (Rick) Cortez et al.,

M. compacta (G. Cunn.) Kreisel & Dring, M. fuliginea (Berk. & M.A. Curtis)

Kreisel & Dring, M. nuda Alfredo & Baseia, M. stercoraria P. Ponce de Leon,

M. rimosa Baseia & Alfredo, M. sulcatostoma C.R. Alves & Cortez, and

M. velutina (Berk. & M.A. Curtis) Kreisel & Dring by its tuberculate

exoperidium (Ponce de Leén 1971; Cortez et al. 2007; Alfredo et al. 2012, 2014;

Alves & Cortez 2013, 2014). The presence of an eucapillitium in M. pyriformis

(Schaeff.) Kreisel & D. Kriiger clearly separates this species from M. puiggarii.

The paracapillitium of M. samoensis (Bres. & Pat) P. Ponce de Leon and

M. subincarnata (Peck) Kreisel & Dring is wider than of M. puiggarii (Ponce

de Leon 1971). Morganella puiggarii forms a sister taxon to M. purpurascens,

the latter having a pitted endoperidium (Ponce de Leén 1971). Moreover,

M. puiggarii was previously known from Brazil and Paraguay (Ponce de Leon

1971).

The combination of morphological and molecular features supports

C. holothurioides and M. puiggarii as new records for Thailand. Prior to this

study, there have been no reports of Calvatia “sect. Sporocristata” and only

four Morganella species (M. compacta, M. fuliginea, M. purpurascens and

M. pyriformis) recorded from the country (Phanichapol 1968, Ellingsen 1982,

Kasuya et al. 2006, Chandrasrikul et al. 2011, Kumla et al. 2014).

Calvatia & Morganella spp. new for Thailand ... 105

Acknowledgements

This work was supported by grants from Chiang Mai University and Thailand

Research Fund, Research-Team Association Grant (RTA5880006). We also thank Dr.

Eric H.C. McKenzie and Dr. Steven L. Stephenson for their helpful comments and

careful review of this article.

Literature cited

Alfredo DS, Leite AG, Braga-Neto R, Baseia IG. 2012. Two new Morganella species from the Brazilian

Amazon rainforest. Mycosphere 3: 66-71. http://dx.doi.org/10.5943/mycosphere/3/1/8

Alfredo DS, Accioly T, Baseia IG. 2014. Morganella arenicola, a new species record from North

and Northeast Brazil. Turkish Journal of Botany 38: 595-599.

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MY COTAXON

January-March 2017— Volume 132, pp. 107-123

http://dx.doi.org/10.5248/132.107

Cyanodermella asteris sp. nov. (Ostropales)

from the inflorescence axis of Aster tataricus

LINDA JAHN” , THOMAS SCHAFHAUSER?’, STEFAN PAN’,

TILMANN WEBER”’, WOLEGANG WOHLLEBEN’, DAVID FEWER?,

KAARINA SIVONEN?, LIANE FLOR‘, KARL-HEINZ VAN PEE‘,

THIBAULT CARADEC®, PHILIPPE JACQUES, MIEKE M.E. HurjBERS®,

WILLEM J.H. VAN BERKEL® & JUTTA LUDWIG-MULLER™

Institut fiir Botanik, Technische Universitat Dresden, 01062 Dresden, Germany

? Mikrobiologie und Biotechnologie, Interfakultares Institut fiir Mikrobiologie und Infektionsmedizin,

Eberhard Karls Universitat Tiibingen, Auf der Morgenstelle 28, 72076 Tiibingen, Germany

> Microbiology and Biotechnology Division, Dept. of Food and Environmental Sciences,

University of Helsinki, Viikinkaari 9, FIN-00014, Helsinki, Finland

‘Allgemeine Biochemie, Technische Universitat Dresden, 01069 Dresden, Germany

° Laboratoire ProBioGEM, Université Lillel- Sciences et Technologies,

Villeneuve d'Ascq, France

° Laboratory of Biochemistry, Wageningen University,

Dreijenlaan 3, 6703 HA Wageningen, The Netherlands

7moved to: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark,

Kemitorvet Bygning 220, 2800 Kgs. Lyngby, Denmark

’ moved to: Gembloux Agro-Bio Tech, Université de Liege,

Passage des Déportés 2, 5030 Gembloux, Belgium

* moved to: Department of Biotechnology, Technical University Delft,

Van der Maasweg 9, 2629 HZ Delft, The Netherlands

*CORRESPONDENCE TO: Linda.Jahn@tu-dresden.de, Jutta.Ludwig-Mueller@tu-dresden.de

ABSTRACT—An endophytic fungus isolated from the inflorescence axis of Aster tataricus is

proposed as a new species. Phylogenetic analyses based on sequences from the ribosomal

DNA cluster (the ITS1+5.8S+ITS2, 18S, and 28S regions) and the RPB2 gene revealed a

relationship between the unknown fungus and the Stictidaceae lineage of the Ostropales.

‘The new species, Cyanodermella asteris, grows in standard fungal growth media as a fluffy,

pink filamentous fungus. Asexual and sexual sporulation has not yet been observed on

media or in the plant.

KEY worDs—Ascomycota, Asteraceae, Lecanoromycetes, Pezizomycotina

108 ... Jahn & al.

Introduction

Aster tataricus (Asteraceae) is native to northern Asia in Siberia,

Mongolia, Japan, Korea, and China. Its roots are well known in traditional

Chinese and Japanese medicine due to their diuretic, antibacterial, antiviral,

and anti-ulcer activities (Shao et al. 1997a, b; Shirota et al. 1997). Diverse

secondary metabolites have been identified in A. tataricus roots: shionone-

type triterpenes [e.g., astataricusones and astataricusol (Zhou et al. 2013);

astershionones (Zhou et al. 2014)], cyclopeptides[e.g., astins (Morita et al.

1996, Xu et al. 2013)], and flavonoids [e.g., quercetin and kaempferol (Wang

et al. 2003)].

With over 2700 species in eleven families, the Ostropales (Hibbett

et al. 2007) represent a large part of the Lecanoromycetes (Ascomycota,

Pezizomycotina). Although most ostropalean fungi are lichenized, parasites

and saprotrophs are also known (Grube & Hawksworth 2007; Lutzoni et

al. 2004; Sherwood 1977a, 1977b; Wedin et al. 2004, 2006). Baloch et al.

(2010), who outline the current taxonomy of the Ostropales, note that most

Stictidaceae are saprotrophs and form very small fruiting bodies.

Here we describe our isolation of an endophytic fungus from the

inflorescence of Aster tataricus and demonstrate its placement in the

Ostropales. DNA sequences from the newly discovered fungus matched no

previously sequenced species, although it did show phylogenetic affinities

with Cyanodermella. For that reason, we propose this endophyte as a new

species, named here Cyanodermella asteris.

Materials & methods

Chemicals were purchased from Duchefa (Haarlem, The Netherlands) and Roth

(Karlsruhe, Germany). Plant hormones were obtained from Duchefa and Sigma-

Aldrich (Hamburg, Germany) and PCR reagents from Thermo Fisher Scientific

(Waltham, USA).

Collecting & field sites

The Aster tataricus host plant was obtained from Sarastro Stauden (Ort im Innkreis,

Austria) and cultivated in the Plant Physiology laboratory greenhouse, Technische

Universitat Dresden, Germany. The cultivar is henceforth referred to as A. tataricus cv.

Austria.

Isolation

The fungus was isolated from surface-sterilized inflorescence axes of Aster

tataricus cv. Austria. After the axes were treated first for 30 s with 70% ethanol +

0.1% Triton X-100, then for 5-7 min with 4.2% sodium hypochlorite + 0.1% Triton

X-100, and washed 3 times with autoclaved distilled water, samples were cultivated

Cyanodermella asteris sp. nov. (Ostropales) ... 109

1 month

——>

Fic. 1. Isolation of the new endophyte Cyanodermella asteris. a. During the first few weeks, the

new endophyte C. asteris was isolated from sterile plantlets growing on hormone containing

MS medium. b. Antimycotic treatment did not reduce the fungal outgrowth, and the fungus

still appeared on the plant surface and grew on the MS medium. c. Isolation and cultivation on

potato dextrose agar produced a fluffy pink filamentous fungus.

on Murashige-Skoog (MS) medium (Murashige-Skoog 1962; 4.4 g/l MS medium

including vitamins, 3% sucrose, 1% phyto agar, pH 5.8) including auxins and/or

cytokinins (ie., naphtylacetic acid, indole-3-butyric acid, 6-benzylaminopurine)

usually used to stimulate sterile plants in vitro. The MS plates were incubated in

long day cycles (16 h light, 23°C; 8 h dark, 18°C). After several transfers of sterile

plantlets to fresh MS plates, we found a fungus growing from the small plants

into the MS medium. Initially considered a contaminant, the fungus was treated

with antimycotics (50 ug/ml Nystatin® for 4 months and penicillin/ streptomycin/

amphotericin B solution for 1 month). However, the treatment did not eliminate the

fungus from the plant culture. In order to identify the fungus, we isolated hyphae

from the plantlets for cultivation on potato dextrose (26.5 g/l potato dextrose

bouillon, 1% phyto agar, pH 5.8) and malt extract agar (3% malt extract, 1% phyto

agar, pH 5.8) in an incubator in the dark at 28°C (Fia.1).

DNA extraction, PCR amplification, and sequencing

DNA was extracted from mycelium grown in potato dextrose broth following

a modified protocol by Mller et al (1992) and precipitated with isopropanol. The

mixture was left overnight at -20°C, spun down, and the DNA pellet washed three

times with 70% ethanol to ensure salt-free DNA. The DNA was amplified using

Phusion’ polymerase according to the LifeTechnologies instruction manual. The

primers (0.5 uM) used for both PCR and sequencing were ITSIF & ITS4 and NS3

& NS8 (White et al. 1990), LROR & LR7 (Vilgalys & Hester 1990), and FRPB2-5F,

-7CR, -7cK, & -114R (Liu et al. 1999). PCR conditions were initial denaturation at

98°C for 30 s, 35 cycles of 98°C for 10 s, appropriate annealing temperature for 30 s,

and 72°C for 1 min followed by a final elongation at 72°C for 5 min. Amplification

was carried out in an Eppendorf Mastercycler ep gradient S. PCR products were

separated on a 1xTAE gel, cut out, extracted using a GE Healthcare Gel Extraction

Kit, and sequenced at Eurofins MWG (Ebersberg, Germany).

110... Jahn & al.

Total DNA was isolated from plants as described above and checked with the

appropriate fungal primers under the same PCR conditions except for an annealing

temperature of 68°C. The PCR fragments were also sequenced and compared with

the known rDNA sequence of the endophytic fungus to confirm the presence of the

fungus in the plants.

Phylogenetic analysis

Sequences of the nuclear internal transcribed spacer region (ITS, including

ITS1+5.8S rDNA+ITS2), the large (28S) and small (18S) nuclear rDNA subunits, and

the second largest RNA polymerase II (RPB2) subunit from Cyanodermella asteris

were run in a BLASTn search (Altschul et al. 1990). The different DNA regions were

also used to build a phylogeny including closely related fungi. For this, the ITS, 18S,

28S and RPB2 sequences from C. asteris and other fungi representing the Stictidaceae

(TABLE 1) were aligned using the Guidance2 Server (Landan & Graur 2008, Penn et

al. 2010, Sela et al. 2015) with the default settings (including MAFFT algorithm) and

Phlyctidaceae and Coenogoniaceae species as out-group. After sequence alignment,

phylogenetic trees were generated from single region and combined sequences

using MEGA6.0 (Tamura et al. 2013). The phylogenetic trees were inferred using

the Maximum Likelihood method based on the General Time Reversible (Nei &

Kumar 2000) or the Kimura-2-parameter model (Kimura 1980). Initial tree(s) for

the heuristic search were obtained by applying the Neighbor-Joining method to a

matrix of pairwise distances estimated using the Maximum Composite Likelihood

(MCL) approach. A discrete Gamma distribution was used to model evolutionary

rate differences among sites. The bootstrap method was used with 1000 replicates

(Felsenstein 1985).

The combined alignment and phylogenetic tree do not include all species, because

rDNA cluster sequences were incomplete for some (TABLE 1).

Microscopy

The fungus was examined under an AP-8 binocular microscope (Thalheim Spezial

Optik, Pulsnitz, Germany) and tissue samples were inspected with transmitted light

with an Axiolab microscope (Zeiss, Jena, Germany). Additionally, fungal hyphae

from a shake culture were stained with 1 g Congo Red in 100 ml 5% SDS to visualize

fungal cell walls and septa (Clémen¢on 2009) under the microscope.

Growth tests

Malt extract broth (50 ml) was inoculated with 0.5 ml of homogenized hyphae of

Cyanodermella asteris and incubated at 180 rpm. The medium was buffered in the

pH test with 50 mM phosphate or 50 mM citrate buffer and cultures were incubated

at 22°C for 10 d. The cultures of the temperature test were incubated in unbuffered

medium (pH 5.3) for 17 d. Growth rates (mm/day) were recorded from 42 d old

cultures on potato dextrose and malt extract agar plates kept at 23°C in the dark.

Colors were coded according to Munsell (1905).

Cyanodermella asteris sp. nov. (Ostropales) ... 111

TABLE 1. DNA sequences from Stictidaceae used for phylogenetic trees

SPECIES

Absconditella

lignicola

A. rubra

A. Sp.

A. sphagnorum

Acarosporina

microspora

Bryodiscus

arctoalpinus

Carestiella socia

Coenogonium

leprieurii

C. luteum

C. pineti

Conotrema

populorum

C. sp.

Cryptodiscus

foveolaris

C. gloeocapsa

C. incolor

C. pallidus

C. pini

C. rhopaloides

C. sp.uncultured

Cyanodermella

asteris

* = ISOLATE

‘ = VOUCHER

*EB211

Svensson & Baloch

SW 187 (S)

Avan den Boom 52517

*Spribille 39168

AGS5_2_17

APalice 3820

*M24

*AFTOL 78

ABaloch SW057

*GG2410

AWedin 7194 (UPS)

AKauft

pa04021998-522

ARyan 31430 (ASU)

*AFTOL 352

AHB Palice

AThor 19164 (UPS)

AUME41471

*GG2610a

AGilenstam 2353 (UPS)

*MW7200

*AN3222

*EBI155

*GG2603a

ABaloch & Arup

SW 166 (S)

*EB93

ATibell 23543 (UPS)

*S:F116574

*EB152

ABaloch & Wedin

SW174 (S)

ABaloch & Arup

SW175 (S)

*EB100

clone F9

strain

03HOR06-2-4

18S

KT454800.1

KRO17250.1

EU940022.1

AY584667.1

AF465 457.1

AF279386.1

U86582.1

FJ904696.1

KT758843

ITS

FJ904669.1

KT454800.1

KRO17125.1

KF128882.1

JX298897.1

DQ782834.1

AY661687.1

HQ650710.1

AY527327.1

AY527313.1

AY527336.1

FJ904673.1

FJ904674.1

NR_121357

FJ904679.1

HM244762.1

FJ904685.1

KP323396.1

KT758843

28S

FJ904669.1

KRO17188.1

AY300825.1

EU940095.1

AY584643.1

HM244760.1

AY661687.1

AF465 442.1

AF279387.1

AY300834.1

AY3008 33.1

AY527313.1

AY527336.1

AY661683.1

FJ904679.1

HM244762.1

KT758843

RPB2

HM244776.1

KRO17501.1

EU940311.1

AY584682.1

HM244781.1

HM244782.1

AY641032.1

AY641038.1

HM244786.1

HM244787.1

HM244788.1

HM244789.1

HM244790.1

HM244791.1

KU934214

112 ... Jahn & al.

SPECIES

C. oleoligni

C. viridula

Glomerobolus

gelineus

Odontotrema

phacidiellum

O. phacidioides

Ostropa barbara

Petractis

luetkemuelleri

P. nodispora

Phlyctis agelaea

P. argena

Schizoxylon

albescens

S. sp.

Stictis brunnescens

S. confusa

S. populorum

S. radiata

S. sp.

S. urceolatum

* = ISOLATE

A = VOUCHER

strain DTO 301-G1

AE. & C. Baloch

SW 129 (S)

AUME29146

*AFTOL 1349

strain JK 5548K

strain JK 5584C

AGilenstam 2625

(UPS)

APalice 11440 (S)

*AFTOL 77

AWedin & Baloch

SW07]1 (S)

ANimis & Tretiach

2000 (TSB 31659)

*AFTOL 381

AGeletti & Tretiach

1995 (TBS)

AA.Orange 17559 (NMW

[C.2007.001.282])

*NMW C.2007.001.284

NPHLAGE08257

*AFTOL 1375

*BP8

A Gilenstam 2696a

(UPS)

AWedin 7919 (UPS)

*GG2365

*GG2359

AWedin 7651 (UPS)

AWedin 7070 (UPS)

AUME41471

*MW7301

AGilenstam 2353 (UPS)

AWedin 7626 (UPS)

APalice (ESS 21520)

*AFTOL 398

AJamie Platt JP222

*GG2445a

*GG2440b

*GG2620b

*AFTOL 96

18S

KX999145.1

U86583.1

DQ247811.1

AY584666.1

AF465 461.1

FJ588712.1

DQ986725.1

DQ401142.1

U86582.1

AY300864.1

DQ983488.1

ITS

KX950434.1

DQ247782.1

HM244773.1

HQ650714.1

NR_120312.1

KJ409433.1

NR_121319.1

AY661689.1

AY661688.1

DQ401143.1

AY527334.1

DQ782846.1

AY527318.1

AY527321.1

AY527332.1

HQ650601.1

28S

KX950461.1

HM244763.1

DQ247798.1

HM244769.1

HM244770.1

AY584642.1

AF465454.1

FJ588713.1

HQ659177.1

DQ986771.1

DQ401144.1

AY661689.1

AY661688.1

DQ401143.1

AY3008 33.1

AF356663.1

AY527318.1

AY527332.1

RPB2

KX999147.1

HM244792.1

DQ247793.1

HM244802.1

HM244803.1

HM244806.1

AY641061.1

KC020280.1

DQ992458.1

HM244813.1

HM244814.1

HM244815.1

HM244817.1

AY641079.1

DQ992478.1

Cyanodermella asteris sp. nov. (Ostropales) ... 113

Taxonomy

Fic. 2. The endophytic fungus Cyanodermella asteris. a, b. Habitus of the host Aster tataricus

showing inflorescences and ground rosette; c. 55 d-old growth on malt extract agar; d. 55 d-old

growth on potato dextrose agar; e. filamentous structure of the colony (36 d old); f. hyphae stained

with Congo Red.

Cyanodermella asteris L. Jahn & Ludw.-Miill. sp. nov. FIG, 2

MycoBAnk MB 814158

Differs from C. viridula by its endophytic habit and its rose-pink hyphae in culture.

TypE—Germany, Saxony, Dresden, isolated as endophyte of Aster tataricus cv. Austria,

21 Mar 2013, L. Jahn, holotype Herb. DR 043292 (metabolically inactive), living culture

at DSMZ under DSM 100826.

EryMoLoGy—tThe epithet ‘asteris’ is derived from the host plant, Aster tataricus cv.

Austria.

Sexual stage unknown. Colonies (14 d old) on potato dextrose agar white

(N 9) to pale pink (2.5R 9/2); in age fading from pale pink to light pink (2.5R

8/6); after 30 days, becoming white towards the margin and turning light and

deep pink (2.5R 6/10) to vivid red (5.0R 4/14) towards the center. Colors less

intense on malt extract agar: 7-14 d old colonies usually colorless (N 9) and

hardly distinguishable from the medium, >50 d old colonies colorless at the

margin and light pink at the center.

114... Jahn & al.

Growth rate 0.7 + 0.3 mm/day on potato dextrose agar and 0.9 + 0.lmm/day

on malt extract agar at an optimum temperature of 22-24°C and a pH of 5.8-6.

HOST & DISTRIBUTION— lhe distribution of the endophytic fungus C. asteris

may be linked to the distribution of its host, A. tataricus, which is native

to northern Asia. Cyanodermella asteris has been isolated and detected by

PCR in several plants of A. tataricus cv. Austria and also detected by PCR in

A. tataricus plants obtained from the Botanical Garden Dresden, Germany,

but which originated in Siberia. Isolation of C. asteris from these plants is

currently in progress.

Phylogenetic results

The nuclear rDNA sequences of Cyanodermella asteris used for identification

(as the “rDNA cluster”) comprised the 18S (1539 bp), ITS (ca. 535 bp) and 28S

(1359 bp) (GenBank KT758843). The first BLASTn search showed a distant

relationship to the Stictidaceae lineage of the ostropalean fungi (TABLE 2). This

led to a second BLASTn search against the Stictidaceae (TABLE 3). The 18S

C. asteris sequence showed as nearest neighbor to C. oleoligni (98% identity)

and C. viridula (95% identity), Acarosporina microspora (94% identity), and

less closely related Stictis species. Similarly, the 28S C. asteris sequence revealed

a relationship with C. oleoligni (94% identity) and C. viridula (95% identity).

The ITS region showed a closest match to C. oleoligni (91% identity), Stictis

sp. (96% identity), and Carestiella socia (96% identity); no ITS sequences were

available for C. viridula.

TABLE 2. BLASTn results of the C. asteris rDNA cluster region

C. ASTERIS BLASTN HITS

Locr # BPs GC % SPECIES ACCESSION # QUERY IDENT E VALUE

COVER

18S 1521 48.0 Cyanodermella oleoligni KX999145.1 74% 98% 0.0

Trapelia involuta AF119499.2 78% 91% 0.0

Trapelia placodioides AF119500.2 78% 95% 0.0

Acarospora smaragdula AY552543.1 78% 91% 0.0

Cyanodermella viridula U86583.1 78% 91% 0.0

ITS 899 53.9 Pleopsidium chlorophanum DQ525472.1 47% 84% 4*10°”

Acarospora insignis LN890273.1 39% 87% 6*10°°

Acarospora smaragdula EU870652.1 38% 86% 4*10°”

28S 1329 49.4 Cyanodermella oleoligni KX950461.1 89% 94% 0.0

Umbilicaria crustulosa HMI161593.1 100% 90% 0.0

Umbilicaria haplocarpa HM161534.1 99% 90% 0.0

Cyanodermella asteris sp. nov. (Ostropales) ... 115

TABLE 3. BLAST» results of the C. asteris rDNA cluster region against Stictidaceae

BLASTN HITS

te SPECIES ACCESSION # tas IDENT E VALUE

Cyanodermella oleoligni KX999145.1 74% 98% 0.0

18S Cyanodermella viridula U86583.1 77% 95% 0.0

Acarosporina microspora AY584667.1 76% 94% 0.0

Stictis urceolatum DQ983488.1 76% 94% 0.0

Cyanodermella oleoligni KX950434.1 32% 91% 210

ie Stictis radiata AY527308.1 30% 88% 3*I.078*

Stictis brunnescens AY661688.1 20% 96% 3*10*

Cryptodiscus pini FJ904682.1 28% 88% 4°10

Carestiella socia AY661682.1 20% 96% 4*10°°

Cyanodermella oleoligni KX950461.1 89% 94% 0.0

Xyloschistes platytropa KJ766680.1 97% 88% 0.0

28S Stictis radiata AY340575.1 77% 90% 0.0

Acarosporina microspore AY584643.1 83% 90% 0.0

Cyanodermella viridula HM244763.1 41% 95% 0.0

69 Absconditella sp. Spribille 39168

7 Abscomditella sphagnorum isolate M24

Phlyctis argena isolate AFTOL 1375

Coenogonium leprieurii voucher K auff pa0402 1998-522

rs) Ostropa barbara isolate AFTOL 77

Sol Stictis urceolatum isolate AFTOL 96

soy Acarosporina microspora isolate AFTOL 78

Glomerobolus gelineus JK 5548K

Conotrema populorum voucher UME4 1471

100! Stictis populorum voucher UME4 1471

Cyanodermella viridula voucher UME29146

Cyanodermella asteris 03 HOR06-2-4

96- Cyanodermella okoligni DTO 301-G1

fk Absconditella rubra voucher van den Boom 52517

fe Stictis radiata voucher Palice (ESS 21520)

= Cryptodiscus gloeocapsa isolate EB93

100) bk Schizoxylon albescens voucher Gilenstam 2696a (UPS)

Petractis nodispora voucher A. Orange 17559 (NMW [C.2007.001.282])

Petractis luetkermuelleri voucher Nimis & Tretiach 2000 (TSB 31659)

Coenogonium luteum voucher Ryan 31430 (ASU)

Fic. 3. 18S sequence-based phylogeny of Cyanodermella asteris and closely related taxa in the

Stictidaceae using Maximum Composite Likelihood based on the Kimura-2-parameter model

(Kimura 1980). The tree with the highest log likelihood (-15822.6420) is shown. A discrete

Gamma distribution was used to model evolutionary rate differences among sites [5 categories

(+G, parameter = 0.6859)]. The percentage of trees clustering the associated taxa is shown next to

the branches. Coenogoniaceae and Phlyctidaceae were used as out-group.

116... Jahn & al.

The RPB2 sequence (2059 bp, GenBank KU934214) was used to refine

phylogenetic relationships within the Stictidaceae. Phylogenetic analyses

based on the individual 18S, ITS, 28S, and RPB2 sequences of Cyanodermella

asteris with other Stictidaceae strongly support a sister-relationship among

C. asteris, C. viridula, and C. oleoligni (Fics 3-6). The combined ML tree also

Stictis populorum isolate MW7301

100} Stictis sp. isolate GG2440b

Conotrema populorum isolate GG2610a

Stictis sp. isolate GG2620b

52 Conotrema sp. isolate AN3222

991 Stictis confiisa voucher Wedin 7070 (UPS)

67| L Carestiella socia isolate GG2410

Stictis brunnescens isolate GG2359

r6| ; Conotrema sp. isolate MW7200

99! Stictis sp. isolate GG2445a

Glomerobolus gelineus isolate AFTOL 1349

Ostropa barbara voucher Wedin & Baloch SW071 (S)

Schizoxylon albescens voucher Gilenstam 2696a (UPS)

1004 Schizoxylon sp. isolate GG2365

Stictis urceolatum isolate AFTOL 96

100

/__-——- Phlyctis argena isolate BP8

Acarosporina microspora isolate AFTOL 78

68 Cyanodermella asteris 033 HOR06-24

Cyanodermella oleoligni DTO 301-G1

Stictis radiata isolate AFTOL 398

99,— Petractis luetkemuelleri isolate AFTOL 381

Petractis nodispora isolate NMW C.2007.001.284

Cryptodiscus rhopaloides voucher Laessoe 12881 (S)

55 Cryptodiscus foveolaris isolate EB155

Cryptodiscus tabularum voucher Gilenstam 2641a (UPS)

92| -— Cryptodiscus gloeocapsa isolate EB93

uncultured Cryptodiscus sp. clone F9

Cryptodiscus pini voucher Baloch & Arup SW175 (S)

9|- Cryptodiscus incolor isolate SF 116574

Cryptodiscus pallidus isolate EB152

100 Absconditella lignicola isolate EB211

Absconditella rubra voucher van den Boom 52517

Absconditella sp. voucher GS5 2 17

100L Absconditella sphagnorum isolate M24

U__. Absconditella sp. Spribille 39168

Coenogonium luteum isolate AFTOL 352

0.5

Fic. 4. ITS sequence-based phylogeny of Cyanodermella asteris and closely related taxa in the

Stictidaceae using Maximum Composite Likelihood based on the General Time Reversible model

(Nei & Kumar 2000). The tree with the highest log likelihood (-11371.6910) is shown. A discrete

Gamma distribution was used to model evolutionary rate differences among sites [5 categories

(+G, parameter = 1.4100)]. The percentage of trees clustering the associated taxa is shown next to

the branches. Coenogoniaceae were used as out-group.

Cyanodermella asteris sp. nov. (Ostropales) ... 117

clusters C. asteris with the other two Cyanodermella species, but the combined

phylogenetic tree is not as well supported as the others due to missing sequences

(Fic. 7; Treebase TB2:S18310).

Discussion

Sequence analyses of the nuclear rDNA cluster from Cyanodermella

asteris implied a close relationship with the Stictidaceae lineage of the

Ostropales (Lecanoromycetes, Pezizomycotina, Ascomycota). The Stictidaceae

are either saprotrophic or lichenized, and no stictidaceous species have been

described as plant endophytes. However, Schulz & Boyle (2005) noted that

Coenogonium leprieurii voucher Kauff pa04021998-522

100 Coenogonium hiteum voucher Ryan 31430 (ASU)

Coenogonium pineti voucher HB Palice

Petractis luetkemuelleri voucher Nimis & Tretiach 2000 (TSB 31659)

Odontrotrema phacidioides voucher Palice 11440 (S)

Odontotrema phacidiellum voucher Gilenstam 2625 (UPS)

Petractis nodispora voucher A. Orange 17559 (NMW [C.2007.001.282])

Ostropa barbara isolate AFTOL 77

Cyanodermella oleoligni DTO 301-G1

Cyanodermella asteris O3HOR06-2-4

6s\— Cyanodermella viridula voucher E. & C. Baloch SW129 (S)

Bryodiscus arctoalpinus voucher Baloch SW057

Acarosporina microspora isolate AFTOL 78

6s Glomerobohus gelineus isolate JK 5584C

99-— Absconditella sp. voucher Palice 3820

76) Absconditella sphagnorum isolate M24

Absconditella sp. Spribille 39168

Absconditella lignicola isolate EB211

Cryptodiscus foveolaris isolate GG2603a

7s| Cryptodiscus pallidus isolate EB152

79L_. Cryptodiscus pini voucher Baloch & Arup SW175 (S)

74, Conotrema sp. isolate MW7200

7| Stictis sp. isolate GG2445a

Stictis brunnescens isolate GG2359

99L Stictis radiata voucher Jamie Platt JP222

Carestiella socia isolate GG2410

gor- Schizoxylon sp. isolate GG2365

Schizoxylon albescens voucher Gilenstam 2696a (UPS)

Stictis sp. isolate GG2620b

Conotrema sp. isolate AN3222

92! | Stictis confusa voucher Wedin 7070 (UPS)

97) Conotrema populorum voucher Gilenstam 2353 (UPS)

Stictis populorum voucher Gilenstam 2353 (UPS)

Phlyctis agelaea

100! Phiyctis argena isolate AFTOL 1375

0.1

Fic. 5. 28S sequence-based phylogeny of Cyanodermella asteris and closely related taxa in the

Stictidaceae using Maximum Composite Likelihood based on the General Time Reversible model

(Nei & Kumar 2000). The tree with the highest log likelihood (-15372.4003) is shown. A discrete

Gamma distribution was used to model evolutionary rate differences among sites (5 categories

[+G, parameter = 0.6449)]. The percentage of trees clustering the associated taxa is shown next to

the branches. Coenogoniaceae and Phlyctidaceae were used as out-group.

118 ... Jahn & al.

$3 Glomerobohus gelineus isolate AFTOL 1349

55 Schizoxylon albescens voucher Wedin 7919 (UPS)

Ostropa barbara voucher Wedin & Baloch SW071 (S)

Carestiella socia voucher Wedin 7194 (UPS)

51 Stictis brunnescens voucher Wedin 7651 (UPS)

Stictis confusa voucher Wedin 7105 (UPS)

100L Stictis populorum voucher Wedin 7626 (UPS)

Acarosporina microspora isolate AFTOL 78

Stictis radiata voucher Jamie Platt JP222

98! Stictis urceolatum isolate AFTOL 96

Absconditella sphagnorum isolate M24

Bryodiscus arctoalpinus voucher Baloch SW057

100 Cyanodermella asteris O3HOR06-2-4

98 Cyanodermella viridula voucher E. & C. Baloch SW129 (S)

Cyanodermella oleoligni DTO 301-G1

36 Absconditella lignicola voucher Svensson & Baloch SW187 (S)

95 Cryptodiscus foveolaris voucher Baloch & Arup SW166 (S)

Cryptodiscus gloeocapsa voucher Tibell 23543 (UPS)

94) Cryptodiscus pallidus voucher Baloch & Wedin SW174 (S)

99_— Cryptodiscus pini voucher Baloch & Arup SW175 (S)

100 Odontotrema phacidiellum voucher Gilenstam 2625 (UPS)

Odontotrema phacidioides voucher Palice 11440 (S)

Cryptodiscus rhopaloides isolate EB100

Petractis luetkemuelleri voucher Geletti & Tretiach 1995 (TBS)

68 Phlyctis agelaea voucher PHLAGE08257

100! Phlyctis argena isolate AFTOL 1375

Absconditella sp. Spribille 39168

Coenogonium pineti voucher Thor 19164 (UPS)

69 $3

78 Coenogonium leprieurii voucher Kauff pa04021998-522

69) Coenogonium luteum voucher Ryan 31430 (ASU)

Fic. 6. RPB2 sequence-based phylogeny of Cyanodermella asteris and closely related taxa in

the Stictidaceae using Maximum Composite Likelihood based on the General Time Reversible

model (Nei & Kumar 2000). The tree with the highest log likelihood (—29390.8544) is shown.

A discrete Gamma distribution was used to model evolutionary rate differences among sites

[5 categories (+G, parameter = 0.5486)]. The percentage of trees clustering the associated taxa

together is shown next to the branches. Coenogoniaceae and Phlyctidaceae were used as out-

group.

many endophytes are facultative in that they adopt a saprotrophic strategy

as soon as their host plant dies. Although C. asteris did not visibly develop

any fruit bodies or other structures on dying and dead plant material of Aster

tataricus, a saprotrophic habit cannot be excluded.

The species within the Stictidaceae live in different ecological niches over

the whole world (see Sherwood 1977a, b, Eriksson 1967, 1981, Wedin et al.

2006, Czarnota, Kukwa 2008, Baloch et al. 2009; Fic. 8). Nonetheless, no

Stictidaceae have been recorded from Siberia or Mongolia. Our species is

Cyanodermella asteris sp. nov. (Ostropales) ... 119

Conotrema populorum voucher Gilenstam 2353 (UPS)

Stictis confusa voucher Wedin 7070 (UPS)

88] ' Conotremasp. isolate AN3222

Conotrema populorum isolate GG2610a

Stictis sp. isolate GG2620b

Carestiella sociaisolate GG2410

Glomerobolus gelineus isolate AFTOL 1349

62 Ostropa barbara voucher Wedin & Baloch SW071 (S)

Schizoxylonsp. isolate GG2365

5 Ostropa barbara isolate AFTOL 77

Stictis urceolatum isolate AFTOL 96

go, Conotremasp. isolate MW7200

Stictis sp. isolateGG2445a

8%] _, StictisbrunnescensisolateGG2359

98 Stictis radiata voucher Jamie Platt JP222

Acarosporinamicrosporaisolate AFTOL 78

72 Conotrema populorum voucher UME41471

Bryodiscusarctoalpinus voucher Baloch SW057

98) AbsconditellalignicolaisolateEB211

Absconditellarubra voucher van den Boom 52517

g2/- Cryptodiscus pallidus isolate EB 152

Cryptodiscus pini voucher Baloch & Arup SW175(S)

Cyanodermella viridula voucher UME29 146

CyanodermellaoleoligniDTO301-G1

Cyanodermella asteris 03HOR06-2-4

Cyanodermella viridula voucher E. & C. BalochSW129(S)

Absconditellasp. Spribille39168

% Absconditellasphagnorum isolate M24

Odontotrema phacidiellum voucher Gilenstam 2625 (UPS)

Odontotrema phacidioides voucher Palice 11440(S)

Petractis nodispora voucher A. Orange 17559

Cryptodiscusrhopaloides isolate EB 100

Cryptodiscus gloeocapsa isolate EB93

56! Schizoxylonalbescens voucher Gilenstam 2696a(UPS)

Coenogonium leprieurii voucher Kauff pa0402 1998-522

3 Coenogonium luteum voucher Ryan31430(ASU)

Fic. 7. Combined rDNA cluster + RPB2 sequence-based phylogeny of Cyanodermella asteris

and closely related taxa in the Stictidaceae using Maximum Composite Likelihood based on the

General Time Reversible model (Nei & Kumar 2000). The tree with the highest log likelihood

(-50824.7554) is shown. A discrete Gamma distribution was used to model evolutionary

rate differences among sites [5 categories (+G, parameter = 0.6150)]. The percentage of trees

clustering the associated taxa together is shown next to the branches. Coenogoniaceae were used

as out-group.

both the first to inhabit a plant with origins in northern Asia and the first

isolated endophyte representing the Stictidaceae.

It should be noted that within the Ostropales, the Stictidaceae are poorly

sampled with relatively few sequences available to generate a phylogeny. The

nearest neighbors of C. asteris were identified as C. viridula (Berk. & M.A.

Curtis) O.E. Erikss. and C. oleoligni van Nieuwenhuijzen & Samson. The only

sequences available for C. viridula—the 18S and 28S—show a 94% identity

match with those from C. asteris. The more diverse ITS region—usually used

120 ... Jahn & al.

Fic. 8. Worldwide distribution of the Stictidaceae and Aster tataricus (Sherwood 1977a,

b, Eriksson 1967, 1981, Wedin et al. 2006, Czarnota, Kukwa 2008, Baloch et al. 2009, Flann

2009). Aster tataricus is native to northern Asia including Siberia, Mongolia, Japan, Korea, and

northern China, from which only a few Stictidaceae are described. Legend: Areas with >15 (®),

10-14 (@), 5-9 (@), or 1-4 (®) different species of Stictidaceae; native distribution of Aster

tataricus (® ); overlapping areas (®) and (®) are coded (®).

to identify and classify unknown fungal species (Schoch et al. 2012)—is not

yet published from C. viridula. Cyanodermella oleoligni shows slightly higher

similarities with C. asteris than C. viridula.

Cyanodermella is characterized by its perithecoid apothecia (Héhnel

1919; Eriksson 1967, 1981), which are very small and closed (Baloch et al.

2010). Only three species are described so far: C. oleoligni, C. viridula, and

C. candida (Setch.) O.E. Erikss.

Cyanodermella viridula is known from old leaves of Leymus arenarius on

beaches found in Sweden and western Russia near St. Petersburg as well as on

twigs in North America, while C. candida is known only on ferns in Mexico

(Eriksson 1967, 1981). Cyanodermella oleoligni (van Nieuwenhuijzen et al.

2016), which was isolated from oil-treated timber of Pinus sylvestris, grows

slowly in grey to green colonies on fungal media. With the sexual stage of

C. asteris not yet known despite an intensive search for ascomata on living

and dead plant material, we expect its classification to be refined as more

sequences from the Stictidaceae become available.

Cyanodermella asteris sp. nov. (Ostropales) ... 121

The role of C. asteris in or for the host plant is not yet clear. Because no

endophyte-free cultivars of A. tataricus are available to us, we are unable to

determine whether the fungus aids the plant’s growth in the environment.

Schulz & Boyle (2005) and Rodriguez et al. (2009) report that many

endophytic fungi do not promote growth even when the plant is under stress.

We speculate that if host plant serves as a habitat to protect C. asteris from the

environment, the need for the fungus to adjust to the host plant might explain

the initially slow growth (later somewhat accelerated; data not shown) of

C. asteris in culture media. Future research will enable us to determine which

metabolic and other features are characteristic for C. asteris.

Acknowledgments

This work was supported by the ERA-IB project MESIAB (EIB 10.004). KH van Pée,

L Flor, J Ludwig-Miller and L Jahn were funded by the Sachsische Staatsministerium

flr Wissenschaft und Kunst (SMWK) and the Sadchsische Aufbaubank (SAB).

W Wohlleben and T Schafhauser were funded by the Bundesministerium fir Bildung

und Forschung (BMBF, grant 0315934). T Weber is supported by a grant of the Novo

Nordisk Foundation. MME Huijbers and WJH van Berkel were supported by grants

from the Netherlands Organisation for Scientific Research (NWO) and The Graduate

School VLAG (Wageningen, The Netherlands). D Fewer and K Sivonen were funded

by Finnish Funding Agency for Technology and Innovation (TEKES 40007/11).

We gratefully acknowledge the revision of the phylogenetic trees by Dr. A Yurkov

(DSMZ, Braunschweig, Germany) as well as Drs. Robert Licking (Botanischer

Garten und Botanisches Museum Berlin, Germany) and Peter Johnston (Landcare

Research, New Zealand) for their expert assistance in revision.

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MY COTAXON

January-March 2017— Volume 132, pp. 125-140

http://dx.doi.org/10.5248/132.125

Eight Caloplaca species newly recorded from Bolivia,

including C. crocina comb. nov.

KARINA WILK*& ADAM FLAKUS

Laboratory of Lichenology, W. Szafer Institute of Botany, Polish Academy of Sciences,

Lubicz 46, Krakow PL-31-512, Poland

* CORRESPONDENCE TO: k.wilk@botany.pl

ABSTRACT— Eight species of Caloplaca s. lat. are reported as new to Bolivia: Caloplaca baueri,

C. cinnabarina, C. crocina, C. darbishirei, C. ochraceofulva, C squamosa, C. subsoluta, and

C. texana. Caloplaca texana is also reported as new for Peru and the Southern Hemisphere.

The new combination Caloplaca crocina is proposed and compared with other Caloplaca spp.

producing hourglass-shaped ascospores. Taxonomic notes and data on the distribution and

habitat preferences are presented for all treated species.

Key worps—lichenized fungi, Ascomycota, Lecanoromycetes, Teloschistaceae, tropical dry

forest

Introduction

Classification of the family Teloschistaceae has been the subject of

intensive research over the last few years (e.g. Gaya et al. 2012, Arup et

al. 2013, Kondratyuk et al. 2013). In the modern classification proposed

by Arup et al. (2013), 39 genera in Teloschistaceae, with more than 1000

species estimated. The former Caloplaca, the most species-rich genus within

Teloschistaceae, has been split into many genera based on phylogenetic

DNA analyses. Research on Teloschistaceae systematics is still in progress

and authors themselves (Arup et al. 2013, Kondratyuk et al. 2013) agree that

the taxonomy is still not settled. As South American Caloplaca species are

poorly known, especially at the molecular level, we decided not to apply the

new systematics in the current paper and temporarily retain the species in

Caloplaca s. lat. (see also note in Materials & methods).

126 ... Wilk & Flakus

Teloschistaceae are represented by five genera in Bolivia: Caloplaca,

Josefpoeltia, Teloschistes, Xanthomendoza, and Xanthoria (Feuerer 2015). Only

six Caloplaca taxa have been reported from the country: C. brebissonii (Fée) J.

Sant. ex Hafellner & Poelt [= Lecanora brebissonii], C. cerina var. chloroleuca

(Sm.) Th. Fr., C. cirrochroa (Ach.) Th. Fr. [= Placodium cirrochroum|], C. crocea

(Kremp.) Hafellner & Poelt subsp. crocea [also as Callopisma xanthaspis],

C. holocarpa (Ach.) A.E. Wade s. lat. [also as Lecanora pyracea], and

C. quadrilocularis (Nyl.) Zahlbr. [= Lecidea quadrilocularis] (Nylander 1859,

1861; Malme 1926; Hafellner & Poelt 1979; Feuerer et al. 1998; Rodriguez

Flakus et al. 2014). There are a few South American countries in which the

knowledge of Caloplaca is relatively comprehensive, e.g., Argentina (57

species), Brazil (52), Chile (45), and Uruguay (30) (Feuerer 2015). Based on

checklists by Feuerer (2015), there are c. 140 Caloplaca species in all of South

America.

Caloplaca s. lat. is fairly well understood in Europe, Asia, North America,

Australasia, and both polar regions, while it remains poorly understood in

Africa and South America. However, data on South American Caloplaca are

included in several taxonomic studies (e.g., Zahlbruckner 1917, 1924; Malme

1926, Magnusson 1950; Hafellner & Poelt 1979;, Karnefelt 1990; Wetmore &

Karnefelt 1998, 1999; Karnefelt et al. 2002; Scutari et al. 2002; Rosato & Arup

2010; Lumbsch et al. 2011; Sochting & Sancho 2012; Vargas Castillo & Beck

2012; Kondratyuk et al. 2014; Sochting et al. 2014), regional floristic surveys

(e.g. Nylander 1859, 1861; Zahlbruckner 1933; Dodge 1936; Follmann 1967;

Osorio 1983; Feuerer et al. 1998; Aptroot 2015), and checklists (e.g. Calvelo

& Liberatore 2002). The work by Malme (1926) deserves special attention,

since it includes descriptions of 34 South American species and a key for their

identification.

This study is part of a project concerning the taxonomic evaluation of the

genus Caloplaca s. lat. in Bolivia. Our main purpose is to determine taxonomic

diversity and phylogenetic relationships of recognized species. We also intend

to analyze the distribution and habitat preferences of individual species.

This paper covers eight species of Teloschistaceae newly recorded for Bolivia:

Caloplaca baueri, C. cinnabarina, C. crocina, C. darbishirei, C. ochraceofulva,

C. squamosa, C. subsoluta, and C. texana. Caloplaca texana is also reported

as new to Peru and the Southern Hemisphere. Caloplaca crocina is compared

with eight Caloplaca species producing ascospores with thickened spore

walls: C. calcitrapa, C. cupulata, C. dichroa, C. haematommona, C. lobulascens,

C. pollinii, C. rheinigera, and C. yuennana. Taxonomic notes and data on the

distribution and habitat preferences for all eight species are presented.

Caloplaca crocina comb. nov. + 8 new records for Bolivia ... 127

Materials & methods

CoLLecTIoNs—The authors examined their own collections and herbarium

specimens from B, KRAM, LD, MEL, S. Both authors collected specimens in the

years 2004-15 throughout Bolivia (depts.: Beni, Chuquisaca, Cochabamba, La

Paz, Potosi, Santa Cruz, Tarija) in the following vegetation zones (Ibisch & Mérida

2004): high mountain vegetation, Tucumano-Boliviano montane forest, semi-desert

Inter-Andean Valleys, Chaquefo and Chiquitano dry forests, savanna Moxos, and

anthropogenic woodlands. Flakus also collected some material from Colca Valley,

Peru. The voucher specimens are available at KRAM and/or LPB.

MORPHOLOGICAL EXAMINATION—The materials were studied using standard

microscopic techniques. Macro-morphological characters were measured from

dry material. Hand-cut sections and squash preparations mounted in water were

examined and measured microscopically, and the granulation of anatomical

structures was observed in polarized light. Solubility of granules/crystals and

color reactions were determined using 25% KOH (K) and 65% nitric acid (N).

Hydrochloric acid (HCI) was used to test the presence of calcium carbonate (CaC0,)

in the substrate.

NOMENCLATURE—In the most recent classification of Teloschistaceae, Arup et

al. (2013) and Kondratyuk et al. (2013) transferred five species treated here to

new genera: Caloplaca darbishirei to Austroplaca Sochting et al.; C. squamosa and

C. subsoluta to Squamulea Arup et al.; C. texana to Wetmoreana Arup et al.; and

C. cinnabarina to Brownliella S.Y. Kondr. The other three featured species and some

other species discussed here do not have modern recombinations.

New combination

Caloplaca crocina (Kremp.) K. Wilk & R. Vargas, comb. nov. PLATE 1

MycoBAnk MB 816545

= Lecidea crocina Kremp., Flora 61: 519. 1878.

Type: Argentina, 1873-1874, Lorentz & Hieronymus (M!, holotype).

= Blastenia crocina (Kremp.) Mill. Arg., Rev. Mycol. (Toulouse) 10: 68. 1888.

= Lecanora crocina (Kremp.) Stizenb., Ber. Thatigk. St.

Gallischen Naturwiss. Ges. 1893-94: 235. 1895.

= Callopisma crocinum (Kremp.) Malme, Ark. Bot. 20A(9): 19. 1926.

= Placodium crocinum (Kremp.) Rasanen, Borbasia 1: 129. 1939.

“Caloplaca crocina” Marcelli (as “crocinum”), nom. inval.

THALLUS not or sometimes slightly parasitized, thin to endophloeic, or

thick, 60-150 tm thick; crustose, continuous to rimose, forming irregular

patches, creamy, sometimes with greenish tinge, epruinose, smooth to slightly

rough (when parasitized); vegetative propagules absent; prothallus usually

present, conspicuous, black. CorTEx poorly developed, amorphous, hyaline,

anthraquinones absent, K-, N-; algal layer discontinuous, algae concentrated

128 ... Wilk & Flakus

in groups. APOTHECIA always present, abundant, scattered or crowded in

small groups of 2-3, round, angular or flexuous when compressed, adnate,

zeorine to almost biatorine, 0.1—-1.0(—1.4) mm wide; disc first concave, then

+plane, rarely slightly convex in old apothecia, orange to brownish orange

or olive; proper margin swollen and prominent in young apothecia, then

thinning and only slightly prominent, yellow orange, often contrasting

against the disc, epruinose; thalline margin conspicuous or inconspicuous to

almost absent, in young apothecia hidden at base of apothecia or invisible,

in mature and old apothecia generally better developed, although often

remaining +hidden at base of apothecia, fragmented (as tiger stripes).

PARATHECIUM thick, <200 um, prosoplectenchymatous and composed of

irregularly orientated hyphae (chondroid hyphae), anthraquinones present;

amphithecium usually strongly reduced, algae scarce and in distinct groups

(only occasionally abundant), cortex absent or amorphous, anthraquinones

absent; epihymenium yellow red or yellow brown, granular (anthraquinones),

K+ purple; hymenium (65-)75-100(-120) um thick; hypothecium yellowish

or pale brownish, K-, oil droplets present or absent, prosoplectenchymatous,

stipe present. PARAPHYSES simple or forked, 1.5-2.0 um broad with upper cell

not or slightly wider, 2-3 um, oil droplets absent. Asci 8-spored. ASCOSPORES

hyaline, hourglass-shaped, 15-24 x 7-10(-13) um, septum (3-)4-5(-6) um,

apical spore walls 1.5-3.0 um.

PYCNIDIA not observed.

SPECIMENS EXAMINED: Caloplaca crocina—BOLIVIA. Dept. BENI, Prov. Ballivian:

near Reyes village, Moxos savanna, 14°18’10”S 67°18’49”W, alt. 192 m, on tree bark,

29 Nov. 2004, Wilk 2507 (KRAM), Flakus 4380 (KRAM, LPB), and 5 Dec 2004, Wilk

2320 (KRAM, LPB); Reyes village, 14°17’54”S 67°20’05’W, alt. 189 m, on wooden fence,

good insolation, 29 Nov. 2004, Wilk 2332 (KRAM). Prov. Cercado, Trinidad, Missones

Guarayos, alt. 250 m, Sept. 1926, E. Werdermann 2490 (S, B). Dept. La Paz, Prov.

Franz Tamayo, Parque National y Area Natural de Manejo Integrado Madidi, Mojos,

along the path to Virgen del Rosario, 14°33’22”S 68°52’27”W, alt. 1635 m, submontane

anthropogenic woodlands, sunny and dry place, 31 Oct. 2007, Wilk 9356, 9380 (LPB),

9367, 9370, 9478 (KRAM). DEPT. SANTA Cruz, Prov. Cordillera, Parque Nacional y

Area Natural de Manejo Integrado Kaa-lya del Gran Chaco: gasoducto Bolivia-Brasil,

between Tucavaca colony and National Park border, 18°31’40”S 60°41’26’W, alt. 314 m,

Chiquitano forest, 4 Dec. 2010, Flakus 19218 (KRAM, LPB); gasoducto Bolivia-Brasil

KP 164, 60 km of Tucavaca colony, 18°27’29”S 61°23’01”W, alt. 292 m, Chaqueiio forest,

3 Dec. 2010, Flakus 18960 (LPB), 18961 (KRAM); Charrata village near campamento

de guardaparques, 18°28’05’’S 62°05’43”W, alt. 307 m, Chaquenio forest, 2 Dec. 2010,

Flakus 20144 (KRAM, LPB). Prov. Chiquitos, near Santa Cruz de la Vieja, mirador a San

José de Chiquitos village, 17°52’21”S 60°45’41”W, alt. 501 m, Cerrado de la Chiquitania

(wooded savannas), 5 Dec. 2010, Flakus 19280 (LPB). Dept. Tarija, Prov. Aniceto

Arce, Serrania de Propiedad Arnold, 22°13’19”S 64°33’41”W, alt. 1309 m, Tucumano-

Boliviano montane forest, 24 Dec. 2010, Flakus 18866/1 (LPB). DEPT. CHUQUISACA,

Prov. Luis Calvo, Parque Nacional y Area Natural de Manejo Integrado Serrania del

Caloplaca crocina comb. nov. + 8 new records for Bolivia ... 129

PiaTE 1. Caloplaca crocina: A—habit (Wilk 2507, KRAM); scale bar= 1 mm. B—ascospores

(Wilk 9380, LPB); scale bar= 10 um. C—vertical section of apothecium (Wilk 9478, KRAM);

scale bar= 200 um. D—prosoplectenchymatic parathecium with irregularly orientated hyphae

(Wilk 9380, LPB); scale bar= 100 um.

Ifao, close to Ticucha, 19°37’26”S 63°50’55”W, alt. 1040 m, disturbed Sub-Andean

Boliviano-Tucumano forests with Acacia, 18 July 2015, Flakus 26310 (KRAM, LPB).

ARGENTINA. Prov. CorRIENTES, Dept. Concepcion, 11 km N de Santa Rosa, 24 Jan.

1974, M.M. Arbo 504 (S).

Caloplaca letrouitioides—AUSTRALIA. Victoria, Gippsland Plain, Gunnamatta

Beach, 1976, R.B. Filson 15871 (MEL, holotype).

ComMMENTS— The combination Caloplaca crocina was suggested by Marcelli in

the original version (dated 2002, unpublished) of the Brazil lichen checklist,

as “Caloplaca crocinum [sic] (Krempelhuber) nao comb. [i.e., not combined]”.

After checking subsequent Marcelli publications and making wide enquiries

amongst lichenologists in South America and internationally, we have found

no evidence that this name was ever validated; Feuerer (2015) cited the species

only as Blastenia crocina.

Caloplaca crocina is diagnosed by its continuous to rimose creamy thallus,

zeorine apothecia with orange discs, and hourglass-shaped ascospores [sensu

130 ... Wilk & Flakus

Navarro-Rosinés et al. 2000; spore walls 1-2(-3) um thick]. The apothecial

thalline margin is usually much reduced and its fragments are visible at

the base of apothecia. In contrast, the proper margin is thick and swollen,

similar to that of the biatorine apothecia in Letrouitia. Caloplaca crocina is

superficially very similar to Letrouitia, which differs in its multi-septate to

muriform spores. Two other Caloplaca species resemble Letrouitia spp.:

Caloplaca letrouitioides S.Y. Kondr. et al. and C. kiewkaensis Yakovczenko et

al. (Kondratyuk et al. 2011). Caloplaca letrouitioides differs from C. crocina

in having truly biatorine apothecia with a more swollen proper margin

that partially overlaps the disc, apothecial discs at first distinctly concave,

and thin-walled ascospores. Caloplaca kiewkaensis is distinguished by its

smaller ellipsoid to almost spherical spores, a parathecium consisting of

radiating hyphae, and a different Far East Russian distribution. See TABLE 1

for a comparison of C. crocina with Caloplaca species characterized by thick-

walled ascospores—C. calcitrapa Nav.-Ros. et al., C. cupulata Poelt & Hinter.,

C. dichroa Arup, C. haematommona Elix & S.Y. Kondr., C. lobulascens Poelt

& Hinter., C. pollinii (A. Massal.) Jatta, C. rheinigera Elix & S.Y. Kondr., and

C. yuennana (Zahlbr.) Poelt & Hinter.

In Bolivia, C. crocina occurs mainly on tree bark (rarely on wood) in

savannas and relatively dry woodlands (e.g., Chaquefio and Chiquitano

forests, Tucumano-Boliviano forest, and submontane anthropogenic

woodlands) at altitudes between 190 and 1635 m. The species is very frequent

in these areas. Caloplaca crocina occurs in subtropical and tropical regions of

Argentina, Brazil, Paraguay (Malme 1926) and Uruguay (Osorio 1972). This

is the first report from Bolivia.

Other species

Caloplaca baueri (Mill. Arg.) Zahlbr., Denkschr. Kaiserl. Akad. Wiss., Wien, Math.-

Naturwiss. Kl. 83: 192. 1909.

SPECIMENS EXAMINED: Caloplaca baueri—BOLIVIA. Dept. BENI, Prov. Itenez, Cerro

de Oricore by Rio San Martin, 13°19’55”S 63°30'37”W, alt. 240 m, dry forest overgrow

granitic hill (an inselberg), 19 Aug. 2008, Flakus 12680/1 & Rodriguez (KRAM, LPB).

Dept. La Paz, Prov. Mufiecas, Camata village, NE slope, 15°14’34”S 68°45’06’W, alt.

2020 m, open place, on a stone fence, 15 May 2006, Wilk 4436 (KRAM).

Caloplaca subunicolor—BRAZIL. MaTTo Grosso, pr. Cuyaba, 6 June 1894, G. Malme

(LD 1036560).

Caloplaca baueri belongs to the C. cinnabarina group (Wetmore & Karnefelt

1999). It is characterized by its continuous to rimose or only slightly areolate,

smooth and glossy, greenish-yellow to yellow-orange thallus that gradually thins

at the margin, with a minimal yellow prothallus. Apothecia are frequent. The

Caloplaca crocina comb. nov. + 8 new records for Bolivia ... 131

TABLE 1. Comparison of Caloplaca crocina and other Caloplaca spp. that produce

thick-walled ascospores with + reduced lumens

(including typical hourglass-shaped spores)

CHARACTERS

SPORES (um)

SEPTUM (tm)

WALL (um)

APOTHECIA

DISC

PROP MARGIN

K RXN

THAL MARGIN

EXCIPLE

PLECTENCHYMA

HYPH. ORIENT

AMPHITHECIUM

ALGAE

THALLUS

K RXN

MORPHOLOGY

SUBSTRATE

HABITAT

DISTRIBUTION

REFERENCES

crocina

15-24 x

8-11

4-9

1.5-2.5

Zeorine

Brownish orange

Orange

+, purple

+, + fragmented

near base

Parathecium,

thick

Prospo-

Irregular

Weakly dev.

Distinct groups

Creamy

K-

Continuous

to rimose

Bark

(rarely wood)

Dry woodland

SAm—Argentina

Bolivia Brazil

Paraguay

Uruguay

Malme (1926);

this paper

calcitrapa

11.5-15 x

6-8

3-6

1-2

Zeorine

Orange

+, purple

Parathecium

Radiating

Abundant

Yellow-orange,

K+ purple

Granulose-

areolate

Calcareous rocks

Medit. vegetation

EUR—France

Spain Italy;

AFR—Algeria

Morocco

Navarro-Rosinés

et al. (2000)

cupulata

16-21 x

8-13

4-7

Unknown

Zeorine

Brownish to

red-orange

Orange

Unknown

Parathecium

Para-

Abundant

White

K-

Granulose-

areolate to

squamulose

Bark, wood

Arid forests

Nepal

Poelt &

Hinteregger

(1993)

dichroa

12-16.5 x

6-8.5

2.5-4

1-2

Zeorine

Orange

+, purple

Parathecium

Radiating

thick-w

Abundant

Yellow to orange

K+ purple

Areolate,

with blastidia

Calcareous rocks

Sun-exposed sites

Europe

Arup (2006)

(concluded on p. 132)

132 ... Wilk & Flakus

Table 1, concluded

CHARACTERS

SPORES

(um)

SEPTUM(um)

WALL (um)

APOTHECIA

DISC

PROP MARGIN

K RXN

THAL

MARGIN

EXCIPLE

PLECTENCH.

HYPH. ORIENT

AMPHITHEC.

ALGAE

THALLUS

K RXN

Morpu.

SUBSTRATE

HABITAT

DISTRIBUTION

REFERENCES

haematommona

10-13 x

5-7

Biatorine (occ.

zeorine)

yOr to brOr

+ purple

Very thin/

Parathecium

Para-

Conglut.

matrix

+/-

Whitish

Unknown

Continuous,

very thin to

almost absent

Bark on

shrub twigs

Dry

woodland

Australia

Kondratyuk

et al.

2007

lobulascens

9-14 x

5-7

2-5

Unknown

Zeorine

Orange-brown

Unknown

+/evanescent

Parathecium

Para-

Abundant

Whitish

Unknown

Rimose-

areolate,

+ phyllidia

Siliceous rock

Warm

temperate

Nepal

Poelt &

Hinteregger

1993

pollinii

12.5-17 x

7-10

4-7

c.1

Biatorine

Brwn/rust/blk

+ rViol > prp

Parathecium

Radiating,

thick-walled

Continuous

to areolate

Bark, wood

Eur NAm

Asia

Wetmore

1994

rheinigera

15-19 x

7-9

3-7

1-2

Biatorine

Rust-brown

Red-violet

Parathecium

Radiating,

conglut.

Grey

Unknown

Continuous

Bark

on twigs

Dry

woodland

Australia

Kondratyuk

et al.

2007

yuennana

21-30 x

11-14

3-5

unknown

Biatorine

(occ. zeorine)

Olive-black

Red-brown

Unknown

—i +

at base

Parathecium

Radiating

— (rarely +)

(few below)

brW

Unknown

Continuous

to areolate

Picea bark

Temperate

mixed forest

China

Poelt &

Hinteregger

(1993)

Caloplaca crocina comb. nov. + 8 new records for Bolivia ... 133

species resembles the European C. ochracea (Schaer.) Flagey, which produces

spores with four locules, non-corticate thallus, and a habit on calcareous rocks.

Caloplaca baueri is very similar to C. subunicolor (Nyl.) Zahlbr., which differs

in having a more areolate, darker orange thallus and lack of prothallus; the

geographical distribution of both species is also quite different, as C. subunicolor

is centered in Africa (Wetmore & Karnefelt 1999). Moreover, C. baueri may also

be confused with C. cinnabarina, which has a distinctly areolate bright orange

thallus with elongated areoles at the thallus margin. Wetmore & Karnefelt

(1999) and Wetmore (2007) provide a detailed description of C. baueri.

The species occurs in Bolivia on silicate outcrops in dry forest, surrounded

by the humid Amazonian zone. Caloplaca baueri is known from Central and

South America, and the Caribbean region; in South America Wetmore &

Karnefelt (1999) have reported C. baueri; this is the first report from Bolivia.

Caloplaca cinnabarina (Ach.) Zahlbr., Nat. Pflanzenfam., 1(1*): 228. 1908.

EXSICCATI EXAMINED— Wetmore, Telos. Exsicc. 28, 29 (KRAM). Weber, Lich. Exs. 446

(as Caloplaca subnitida) (KRAM).

SPECIMENS EXAMINED—BOLIVIA. Dept. La Paz, Prov. Franz Tamayo, Parque

National y Area Natural de Manejo Integrado Madidi, along the way from Mojos to

Insensio, 14°33’22”S 68°52’27’W, alt. 1635-1960 m, sunny, dry area with sparse

trees and shrubs, 1 Nov. 2007, Wilk 9545 (KRAM, LPB). Dept. SANTA CRUZ, Prov.

Chiquitos, near Santa Cruz de la Vieja, mirador a San José de Chiquitos village,

17°52’21”S 60°45’41”W, alt. 501 m, Cerrado de la Chiquitania (wooded savannas), 5

Dec. 2010, Flakus 19376, Quisbert & Wolski (KRAM, LPB). AUSTRALIA. NEw SouTH

WALES, ‘Bush Bottoms, NE of Goulburn, 18 Oct. 1987, H. Streimann 38875 (B).

Caloplaca cinnabarina is characterized by its bright orange areolate thallus with

the areoles slightly elongated at the thallus margin. The apothecia are more or

less immersed in and concolorous with the thallus. See Wetmore & Karnefelt

(1999), Wetmore (2007) and Joshi et al. (2011) for detailed descriptions. To

distinguish C. cinnabarina from C. baueri see remarks above under that species.

The recently described Australian species, C. brownlieae S.Y. Kondr. et al., differs

from C. cinnabarina in having a dull pink to dull brownish-orange thallus and

wider paraphyses tips, ascospores, and spore septa. According to Lumbsch et

al. (2011), C. brownlieae also differs chemically in having ovoic and lecanoric

acids and a high concentration of gyrophoric acid. Caloplaca rubelliana (Ach.)

Lojka, a primarily European species, is very similar to C. cinnabarina, but is

distinguished by its greyish-orange or brownish-orange thallus that thins at the

margin and scarlet red apothecia; moreover, it does not form elongated areoles

at the thallus margin (Wetmore & Karnefelt 1999).

Caloplaca cinnabarina occurs in Bolivia on silicate rocks in dry woodland

areas. This rather well known species, widespread in subtropical and tropical

134 ... Wilk & Flakus

regions, has been reported from North and Central America (Wetmore &

Karnefelt 1999), Africa (Karnefelt 1988), Australia (Karnefelt 2003), New

Zealand (Galloway 2007), and Asia (Joshi et. al. 2010). In South America it is

known from Argentina (Calvelo & Liberatore 2002), Uruguay (Osorio 1983)

and Venezuela (Aptroot 2015). This is the first report from Bolivia.

Caloplaca darbishirei (C.W. Dodge & G.E. Baker) Cretz., Bul. Grad. Bot. Univ. Cluj

21: 140. 1941.

SPECIMEN EXAMINED—BOLIVIA. Dept. Poros, Prov. Nor Lipez, Pinturas Rupestres

near Mallku Villamar village, 21°46’20”S 67°29’05’W, alt. 4038 m, high Andean area, 6

Dec. 2009, Flakus 14802 (LPB).

The species is characterized by its orange-yellow, squamulose, and sorediate

thallus. The squamules are peltate with incised margins, and the soralia develop

on the underside of the squamule margins. Soredia are concolorous with the

thallus. The closely related Caloplaca soropelta (E.S. Hansen et al.) Sochting

differs in its less incised squamule margins and golden yellowish soredia

distinctly lighter than the thallus. For more information about both species see

Sochting & Castello (2012).

Caloplaca darbishirei occurs in Bolivia on silicate rock together with

C. texana in high montane regions. It is known from Antarctica and the

southernmost part of Argentina in South America (Sochting & Castello 2012).

This is the first report of C. darbishirei from Bolivia.

Caloplaca ochraceofulva (Mill. Arg.) Jatta, Nouv. Giorn. Bot. Ital., n.s. 17: 194.

1910.

EXSICCATI EXAMINED—Almborn, Lich. africani 69 (as C. subnitida) (LD).

SPECIMENS EXAMINED: Caloplaca ochraceofulva—BOLIVIA. Dept. La Paz, Prov.

Bautista Saavedra, Charazani village, alt. c. 3000 m, by the road, open place, 13 May

2006, Wilk 4144 (KRAM). DEPT. COCHABAMBA, Prov. Quillacollo, area of Inkarraya-

Sipesipe, semidesert Inter-Andean Valleys, 17°29’25”S 66°22’09”W, alt. 3146 m, rocky

and shrubby slope, sunny place, E exp., 17 Dec. 2004, Wilk 3277 (KRAM, LPB).

ARGENTINA. Prov. Juyuy, Santa Barbara, alt. 1300 m, 9 July 1901, Rob. E. Fries 50

(LD).

Caloplaca isidiosa—BRAZIL. Banta, zwischen Feira de Santana und Milagres, alt. 200

m, an bodennahen Granitblécken in einer Caatinga, 21 July 1980, K. Kalb [Lichenes

neotropici no. 207] (B).

Caloplaca ochraceofulva is characterized by its placodioid thallus with moderately

long and flat lobes. The central thallus is rimose-areolate, with numerous isidia

produced on the margins of rather thick subsquamulose areoles. The species

usually occurs in the sterile form. For more information see Malme (1926, as

C. subnitida), Karnefelt (1990) and Wetmore & Karnefelt (1998). The similar

Caloplaca crocina comb. nov. + 8 new records for Bolivia... 135

C. texana differs in having the central thallus densely covered by phyllidia.

Caloplaca ochraceofulva may also be confused with C. isidiosa (Vain.) Zahlbr.,

which is distinguished by its coralloid or simple elongate isidia that densely

cover the central thallus and frequent apothecia (Karnefelt 1988, 1990).

Caloplaca ochraceofulva occurs in Bolivia on silicate or slightly calcareous

rocks in high Andean regions at an altitude of c. 3000 m. It has an African-

South American distribution pattern (Karnefelt 1988) and has been recorded

from Argentina (Malme 1926), Brazil (Osorio & Fleig 1990), Peru (Ramos

2014), and southern and eastern Africa, including one locality on the Arabian

Peninsula (Karnefelt 1990). This is the first report from Bolivia.

Caloplaca squamosa (B. de Lesd.) Zahlbr., Cat. Lich. Univ. 10: 629. 1940.

EXxsICCATI EXAMINED— Weber, Lich. Exs. Colo. 414 (as Caloplaca modesta) (KRAM).

SPECIMEN EXAMINED—BOLIVIA. Dept. SANTA CRUZ, Prov. Caballero, area of Siberia

village, 17°49’38”S 64°45’14”W, alt. 3480 m, the field on hill above road (on the fringes

of the Yungas cloud forest), open, foggy and windy area, on sandstone, 15 Dec. 2004,

Wilk 3049 (KRAM, LPB), 3073, 3125 (KRAM).

Caloplaca squamosa and C. subsoluta (discussed below) belong to the

C. squamosa group, which is characterized by a more or less squamulose

thallus, orange apothecia, and a paraplectenchymatous apothecial proper

margin (Wetmore 2003). Both C. squamosa and C. subsoluta are highly variable

and more study is needed to refine their taxonomy. Typical C. subsoluta differs

from C. squamosa by its areolate to subsquamulose thallus lacking short lobes

and smaller apothecia lacking distinct thalline margins. For more information

on both species see Wetmore (2003, 2007).

Caloplaca squamosa occurs in Bolivia on silicate rocks in open anthropogenic

areas close to Yungas cloud forest at an altitude of c. 3000 m. Its centre of

distribution lies in southwestern North America (Wetmore 2003, 2007). In

South America it is known from the Galapagos Islands (Bungartz et al. 2013).

This is the first report from Bolivia.

Caloplaca subsoluta (Nyl.) Zahlbr. s. lat., Cat. Lich. Univ. 7: 185. 1931.

EXSICCATI EXAMINED— Wetmore, Telos. Exsicc. 71, 72, 73, 92 (KRAM). Suza, Lich.

Bohemoslov. 206 (as C. irrubescens) (KRAM).

SPECIMENS EXAMINED—BOLIVIA. DEPT. SANTA CRUZ, Prov. Caballero, area of Siberia

village, 17°49’38”S 64°45’14”W, alt. 3480 m, a field on hill above road (on the fringes of

the Yungas cloud forest), open, foggy and windy area, 15 Dec. 2004, Wilk 3134 (KRAM,

LPB). Dept. COCHABAMBA, Prov. Quillacollo, area of Inkarraya-Sipesipe, 17°29’25”S

66°22'09”W, alt. 3146 m, semidesert Inter-Andean Valleys, rocky and shrubby slope,

sunny place, E exp., 17 Dec. 2004, Wilk 3228 (LPB). PERU. AREQUIPA, Prov. Caylloma,

Valle del Colca valley, near Soccoro village, 15°38’32”S 71°43’22”W, alt. 3349 m, open

semi-desert montane area, 3 July 2006, Flakus 9409 & Cykowska (KRAM).

136 ... Wilk & Flakus

Wetmore (2003) also treats Caloplaca subsoluta in the C. squamosa group. The

species, together with C. americana (Malme) Zahlbr., C. irrubescens (Arnold)

Zahlbr., and C. modesta (Zahlbr.) Fink form a group of very similar species,

but the taxonomic affiliation is unclear. The interpretation of these species

varies according to author; for example, Wetmore (2003) treats C. americana,

C. irrubescens, and C. modesta as synonyms of C. subsoluta. For comparison

with C. squamosa, see comments above under that species.

Caloplaca subsoluta occurs in Bolivia on sandstone rocks in open

anthropogenic areas at altitudes above 3000 m. The species is distributed

worldwide and has been reported from Europe (e.g. Hafellner & Turk 2001),

North America (Wetmore 2003), Asia (e.g. Joshi et al. 2010), and Africa (e.g.

Fryday 2015). In South America it has been recorded from Argentina, Brazil

(Malme 1926, as Callopisma americanum), Peru (Ramos 2014), Uruguay

(Osorio 1983, as Caloplaca americana), Venezuela (Aptroot 2015), and the

Galapagos Islands (Bungartz et al. 2013). This is the first report from Bolivia.

Caloplaca texana Wetmore & Karnefelt. Bryologist 101(2): 247. 1998.

EXSICCATI EXAMINED— Wetmore, Telos. Exsicc. 8, 74, 75 (KRAM). Nash, Lich. Exs. 256

(KRAM).

SPECIMENS EXAMINED—BOLIVIA. Dept. Porosl, Prov. Nor Lipez, Pinturas Rupestres

near Mallku Villamar village, 21°46’20”S 67°29’05’W, alt. 4038 m, high Andean area,

6 Dec. 2009, Flakus 14802/1 (KRAM), Flakus 14812 & Rodriguez (LPB). PERU.

AREQUIPA, Prov. Caylloma, Valle del Colca valley, near Soccoro village, 15°38’32”S

71°43’22”'W, alt. 3349 m, open semi-desert montane area, 3 July 2006, Flakus 9404,

9408, 9411, 9414, 9415 & Cykowska (KRAM).

Caloplaca texana is a distinctive species distinguished by its thick placodioid

thallus that produces moderately long flat marginal lobes +loosely attached

to the substratum. The central thallus is areolate with numerous phyllidia.

The species usually occurs in the sterile form. The similar C. ochraceofulva is

distinguished by its globose isidia on the squamulose areole margins. Caloplaca

texana may be confused with C. appressa Wetmore & Karnefelt, which differs

in its marginal lobes tightly appressed to the rock, frequent reddish apothecia,

and lack of vegetative diaspores (Wetmore & Karnefelt 1998).

Caloplaca texana occurs in Bolivia on silicate rocks in high montane areas

at altitudes above 4000 m. It was originally described from North America,

and these Bolivian and Peruvian records represent the first reports from the

Southern Hemisphere.

Acknowledgements

We are grateful to Mark R.D. Seaward (Bradford, UK), Reinaldo Vargas Castillo

(Santiago, Chile), and Shaun Pennycook for reviewing the manuscript and providing

Caloplaca crocina comb. nov. + 8 new records for Bolivia ... 137

important suggestions and improvements. We thank L. Sliwa and M. Piatek (Krakow,

Poland) for constructive discussion on some taxonomic and nomenclatural issues,

and A. Spielmann (Mato Grosso do Sul, Brazil) and A. Aptroot (Soest, Netherlands)

for help in clarifying the C. crocina nomenclatural status. We thank the staff of

the Herbario Nacional de Bolivia, Universidad Mayor de San Andrés (LPB), for

collaboration and invaluable help during the field studies. The first author is also

grateful to P. Jorgensen (St. Louis, MO, US) for the possibility to collaborate with

the team working within the Madidi Project and to T. Wilk (Krakow, Poland) for

assistance during the fieldwork. The second author wishes to thank P. Rodriguez

Flakus (Krakow, Poland), J. Quisbert (La Paz, Bolivia), A. Wolski (Gliwice, Poland),

and all protected areas staff for their kind help during the fieldwork. We thank the

Curators of B, KRAM, LD, MEL, S for providing the material for study, and A. Aptroot

(Soest, Netherlands), C. Wetmore (Minnesota, US) and R. Vargas Castillo (Santiago,

Chile) for literature support. Financial support was provided to the first author by the

National Science Centre (NCN, grant no. N N303 821740) and statutory funds by the

W. Szafer Institute of Botany, Polish Academy of Sciences.

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MY COTAXON

January-March 2017— Volume 132, pp. 141-148

http://dx.doi.org/10.5248/132.141

Notes on rust fungi in China 3. Puccinia adenocauli comb. nov.

and its life cycle and new host

JING- XIN Jr, Q1 WANG’, ZHUANG LY, Yu LY & MAKOTO KAKISHIMA” 3"

‘Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun, Jilin 130118 China

College of Plant Pathology, Shandong Agricultural University, Taian 271000 China

°University of Tsukuba, Tsukuba, Ibaraki 305-8572 Japan

* CORRESPONDENCE TO: kakishima.makoto.ga@u.tsukuba.ac.jp

AsBsTRACT— The heteroecious and macrocyclic life cycle of Aecidium adenocauli was

proved with field observations and inoculation experiments in China. Its spermogonial

and aecial stages were produced on Adenocaulon himalaicum, and uredinial and telial

stages were produced on Carex onoei, which is a new host for this rust fungus. Uredinial

and telial stages (teleomorph) of this rust fungus were formerly described as Puccinia

carici-adenocauli from Japan. However, the current International Code of Nomenclature

(ICN (Melbourne)) has abolished the distinction between “anamorph” names and

“teleomorph” names, and we propose the new combination Puccinia adenocauli, based

on Ae. adenocauli 1913, which now has priority over P. carici-adenocauli 1999. Its neotype

and epitype are also designated.

Key worps—Asteraceae, Cyperaceae, Pucciniomycetes, taxonomy, Uredinales

Introduction

Aecidium adenocauli has been reported to produce spermogonial and aecial

stages (anamorph) on Adenocaulon himalaicum Edgew. (Asteraceae) from

Japan, Korea, Russian Far East, and China (Sydow & Sydow 1913, Ito 1950,

Hiratsuka 1963, Azbukina 1974, Zhuang 1990, Hiratsuka et al. 1992). However,

Kakishima et al. (1999) showed with inoculation experiments that its uredinial

and telial stages (teleomorph) were produced on Carex hakonensis Franch. &

Sav. and C. uda Maxim. (Cyperaceae) in Japan and described this teleomorph

as a new species, Puccinia carici-adenocauli. Azbukina (2005) also reported

142 ... Ji & al.

this species on Ad. himalaicum and C. uda from the Russian Far East based on

morphological observations.

During investigations of rust fungi in Jilin Province, northeast China, we

found the aecial stage of a rust fungus on Ad. himalaicum. This rust fungus

was morphologically similar to Ae. adenocauli, but its uredinial and telial

stages had not been clarified in China. Therefore, we carried out field survey

and inoculation experiments to determine life cycle of this rust fungus. We

report here the results of inoculation experiments with this rust fungus and

its morphology based on specimens collected in China. We also discuss the

taxonomic treatment of this fungus according to the current International

Code of Nomenclature for algae, fungi, and plants (ICN (Melbourne); McNeill

et al. 2012) and propose the new combination Puccinia adenocauli.

Materials & methods

Field survey and inoculation experiments

An aecial stage on Ad. himalaicum was found in the forest floor at Hongyegu,

Jiaohe, Jilin, Jilin Province, China (43°42’13”N 127°04’18”E, alt. 537m) in June

2015 during a survey of rust fungi. This forest mainly consists of species of Betula,

Populus, and Quercus and is a typical deciduous broad-leaved forest in this area.

Uredinial and telial stages of this rust fungus were suspected to occur on species

of Carex because a heteroecious species, P. carici-adenocauli, alternating between

species of Adenocaulon and Carex, had been reported from Japan (Kakishima et al.

1999) and the Russian Far East (Azbukina 2005). Therefore, we looked for rust fungi

on Carex species in the area where the aecial stage on Ad. himalaicum had occurred

and collected telial stages on C. onoei Franch. & Sav. and C. ussuriensis Kom. in

September 2015 and May 2016. These telial stages were morphologically identified

as belonging to Puccinia and were used for inoculations.

Inoculation experiments were carried out during May-June 2016 at Jilin

Agricultural University, Changchun, Jilin Province. For basidiospore inoculations

the leaves with telia collected on 13 May 2016 (Fic. 2B) were kept in water for 1-2

hours to induce teliospore germination. These leaves were cut into small pieces (ca.

5 mm?) and placed on healthy leaves of Ad. himalaicum, which was transplanted

from the same forest cited above and maintained in plastic pots at Jilin Agricultural

University. No natural infection of the plant from fields was confirmed before

inoculations. The inoculated plants were kept in a moist atmosphere in a plastic box

in darkness at 15-20°C for three days and then transferred to a place near windows

at 15-20°C for observations.

Aeciospores produced on Ad. himalaicum by basidiospore inoculations were then

used as inocula. Aeciospores were dusted onto wet filter papers (about 3 mm”) and

these papers were then placed on young healthy leaves of C. onoei planted in plastic

pots. The plants were kept in a plastic box under the same conditions as basidiospore

inoculations, later transferred to a place near windows.

Puccinia adenocauli comb. nov. (China) ... 143

Morphological observations

Specimens collected in the field or obtained from inoculations were used for

morphological observations. Specimens of Puccinia carici-adenocauli including

the type specimen (TSH-R1693) deposited in the Mycological Herbarium of the

Graduate School of Life and Environmental Sciences, University of Tsukuba,

Tsukuba, Japan (TSH), were borrowed for comparative morphology.

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, sori and spores obtained from dry specimens were attached

to specimen 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 5-7 kV.

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).

Results & discussion

Life cycle

Seven to ten days after inoculations with basidiospores from teliospores on

C. onoei, small yellow spots of spermogonia appeared on the upper leaf surface

of Ad. himalaicum (Fic. 1A). About 7 days later, cupulate aecia with aeciospores

were produced on the lower leaf surface of the same leaves (Fic. 1B). No

infection was observed on Ad. himalaicum in the inoculations with teliospores

from C. ussuriensis; this suggests that the C. ussuriensis rust is probably a

different species from the rust on C. onoei. About 12 days after inoculations with

aeciospores from Ad. himalaicum, pale yellow spots appeared on C. onoei and

then powdery uredinia were produced (Fics 2C, 2E). The inoculation results

confirmed that the rust fungus producing spermogonial and aecial stages on

Ad. himalaicum was conspecific with the rust fungus producing uredinial and

telial stages on C. onoei.

Morphology & taxonomy

Morphological observations using LM and SEM of specimens obtained

from the field and inoculations showed that spermogonial and aecial stages on

Ad. himalaicum and uredinial and telial stages on C. onoei were identical with

the morphology of P. carici-adenocauli (anamorph: Ae. adenocauli) described

by Kakishima et al. (1999). Their identity was also confirmed by comparative

144 ... ji & al.

morphology with specimens from TSH. However, according to McNeill et al.

(2012: Art. 59.1), since 1 Jan. 2013, the earliest legitimate holomorphic name

of this species is Ae. adenocauli, and P. carici-adenocauli is a later synonym

(Kakishima et al. 1999). Therefore, we propose a new name based on the

earlier name. Sydow & Sydow (1913) described Ae. adenocauli based on the

specimen on “Ad. bicolor” [a misdetermination of Ad. himalaicum] collected

at Kuzumaki-machi, Iwate Prefecture, Japan on 6 July 1907. This specimen was

not found in any herbarium, including B (Berlin, Germany), and it is suspected

that the specimen was lost during World War II. Therefore, we designate

a neotype specimen on Ad. himalaicum for this species and also an epitype

specimen on Carex onoei. We also provide a description based on specimens

obtained in China.

Puccinia adenocauli (Syd. & P. Syd.) Jing X. Ji & Kakish., comb. nov. Fics 1-3

MycoBank MB817618

= Aecidium adenocauli Syd. & P. Syd., Ann. Mycol. 11: 111, 1913.

Type: Stages 0, I on Adenocaulon himalaicum, Japan, Yamanashi Prefecture,

Yamanakako-mura, 25 May 1983, leg. M. Kakishima (Neotype designated here,

TSH-R1689). Stage III on Carex onoei, China, Jilin Province, Jilin, Jiaohe, 13 May

2016, leg. J.X. Ji & M. Kakishima (Epitype designated here, HMJAU8275).

= Puccinia carici-adenocauli Kakish., Yokoi & Y. Harada, Mycoscience 40: 506, 1999.

Spermogonia subepidermal (type 4 of Cummins & Hiratsuka 2003). Aecia

subepidermal, erumpent, Aecidium-type with peridia and catenulate spores.

Aeciospores catenulate, globose, obovoid or sometimes angular, 13-18 x

10-15 um (av. 15 x 12 um), walls hyaline, 0.6 um thick, verrucose with large

granules. Uredinia subepidermal, pale yellow, erumpent, without paraphyses.

Urediniospores pedicellate, obovoid or ellipsoid, 21-27 x 16-22 um (av. 23 x

20 um), walls hyaline, echinulate, 1.6-1.2 um (av. 0.9 um), germ pores obscure.

Telia subepidermal, erumpent, dark brown to black. Teliospores 2-celled by

transverse septum, borne singly on pedicels, clavate to oblong with round to

obtuse apex and attenuate toward bases, constricted at septum, 30-47 x 15-26

um (av. 40 x 20 um), walls dark brown, smooth, 0.6-2.0 um (av. 1.2 um) thick,

thickened at apex.

SPECIMENS EXAMINED — CHINA, JILIN PROVINCE: Stages 0, I on Adenocaulon

himalaicum, (Jilin, 24 June 2015, HMJAU8205, cultured at Jilin Agricultural Univ.,

Changchun, 11 June 2016, HMJAU8277, cultured at Jilin Agricultural Univ., Changchun,

24 June 2016, HMJAU8278, Tonghua, 8 June 2016, HMJAU8276); Stage II on Carex

onoei, 2 July 2016, cultured at Jilin Agricultural Univ. (HMJAU8274); Stage III on C.

onoei,13 May 2016, Jilin (HMJAU8275).

Hosts & DISTRIBUTION — Stages 0, I on Adenocaulon himalaicum: Cuina (Zhuang

1990), Korea (Ito 1950, Hiratsuka 1963, Hiratsuka et al. 1992, Kakishima et al. 1999),

RUSSIAN Far East (Azbukina 1974, 2005, Kakishima et al. 1999), Japan (Ito 1950,

Puccinia adenocauli comb. nov. (China) ... 145

Fig. 1. Puccinia adenocauli on Adenocaulon himalaicum: spermogonial and aecial stages.

A. Spermogonia on upper leaf surfaces produced by basidiospore inoculation. B. Aecia on lower

leaf surface produced by basidiospore inoculation. C. Vertical section of a spermogonium.

D. Aeciospores with large granules. E. Catenulate aeciospores surrounded by peridium in a

vertical section of an aecium. Scale bars: C, D = 15 um; E = 20 um.

Hiratsuka et al. 1992, Kakishima et al. 1999). Stages II, III on Carex hakonensis: JAPAN

(Kakishima et al. 1999). II, III on C. uda: Russian Far East (Azbukina 2005), Japan

(Kakishima et al. 1999). Stages II, III on C. onoei: CHINA.

The frequent occurrence of aecial stage on Ad. himalaicum indicates that this

rust fungus is widely distributed in East Asia, although its uredinial and telial

stages on Carex species have been rarely reported. Collection of this rust fungus

on Carex is difficult because Carex species are short and their leaves are very

146 ... Ji & al.

Fig. 2. Puccinia adenocauli on Carex onoei: uredinial and telial stages. A. Carex onoei with

flowers taken on forest floor at Jilin, Jilin Province in May 2016. B. Dark brown telia (T) on

the dead leaves. C. Uredinia (U) on the leaf surface produced by aeciospore inoculation.

D. Echinulate urediniospores obtained by aeciospore inoculation. E. Pale yellow uredinia on

lower leaf surface produced by aeciospore inoculation. F. Vertical section of a uredinium.

G. Teliospores. Scale bars: D = 20 um; F = 50 um; G = 30 um.

thin. Our field observations and inoculations have shown, for the first time,

that C. onoei serves as uredinial and telial host of this rust fungus in China.

Puccinia adenocauli comb. nov. (China) ... 147

Fig. 3. Puccinia adenocauli observed with SEM. A. Aecium. B. Catenulate aeciospores in an

aecium on Adenocaulon himalaicum. C. Verrucose aeciospore with large granules. D. Echinulate

urediniospores. E. Uredinium produced on the leaf surface of Carex onoei. F. Vertical section of

a uredinium. Scale bars: A = 40 um; B = 10 um; C = 5 um, D = 15 um; E = 20 um; F = 50 um.

Carex onoei also represents a new host for this rust fungus. Two other Carex

spp., which are uredinial and telial hosts of this rust, have been reported from

Japan and the Russian Far East (Kakishima et al. 1999, Azbukina 2005). These

three Carex species (C. hakonensis, C. onoei, C. uda) belong to Carex sect. Rarae

and it is, therefore, probable/possible that this rust fungus may be host specific

to this section of Carex for its survival.

148 ... fi&al.

Acknowledgments

This work was financed by the Fungal Flora in 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. R. Liicking (Botanischer

Garten und Botanisches Museum Berlin, Berlin, Germany) for checking specimens

of Ae. adenocauli. We also thank Dr. S. Zhang (Institute of Botany, Beijing, China)

for identification of Carex species.

Literature cited

Azbukina ZM. 1974. Rust fungi of the Soviet Far East. Nauka, Moscow.

Azbukina ZM. 2005. Rust fungi. 1-616, in: ZM Azbukina (ed.). Cryptogamic plants, fungi and

mosses of the Russian Far East, vol. 5. Dalnauka, Vladivostok. (In Russian)

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi, 3"! ed. American Phytopathological

Society, St. Paul, Minnesota.

Hiratsuka N. 1963. A provisional list of Uredinales of Korea. Korean Journal of Microbiology

1: 51-64.

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.

Kakishima M, Yokoi M, Harada Y. 1999. Puccinia carici-adenocauli, a new rust fungus

on Carex, and its anamorph, Aecidium adenocauli. Mycoscience 40: 503-507.

http://dx.doi.org/10.1007/BF02461027

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

Sydow H, Sydow P. 1913. Ein Beitrag zur Kenntnis der parasitischen Pilzflora des nordlichen

Japans. Annales Mycologici 11: 93-118.

Zhuang JY. 1990. Additions to Aecidium from China. Acta Mycologica Sinica 9: 191-195.

MY COTAXON

January-March 2017— Volume 132, pp. 149-152

http://dx.doi.org/10.5248/132.149

Colletotrichum fioriniae comb. & stat. nov.,

resolving a nomenclatural muddle

SHAUN R. PENNYCOOK

Landcare Research Manaaki Whenua

Private Bag 92 170, Auckland, New Zealand

CORRESPONDENCE TO: PennycookS@LandcareResearch.co.nz

ABSTRACT— The widely used name “Colletotrichum fioriniae’, published in 2009, is invalid; it

is neither a valid new combination for “Colletotrichum acutatum var. fioriniae” (itself invalid)

nor a valid new name, because no Latin diagnosis or description was presented or referenced.

With the abolition of “dual nomenclature” (the separate nomenclatures of teleomorphs and

anamorphs) under the current International Code of Nomenclature, it is now possible to

publish Colletotrichum fioriniae as a new combination based on Glomerella acutata vat.

fioriniae, the only validly published name for this taxon.

Key worps—Glomerellaceae, Glomerella fioriniae, Colletotrichum acutatum group A3,

Colletotrichum acutatum group C

Introduction

Glomerella acutata var. fioriniae was proposed with an accompanying

Latin diagnosis by Marcelino et al. (2008), together with an asexual

morph, “Colletotrichum acutatum var. fioriniae”. In 2008, the then current

International Code of Botanical Nomenclature (ICBN (Vienna); McNeill et al.

2006: Art. 59.1) permitted the simultaneous publication of separate names for

the asexual and sexual morphs of a single taxon. However, the lack of a Latin

diagnosis or description means that Marcelino et al’s (2008) Colletotrichum

name is invalid (McNeill et al. 2012: Art 39.1).

Within Colletotrichum acutatum sensu lato, Shivas & Tan (2009) identified

a separate clade (“formerly C. acutatum group C [of Lardner et al. 1999] or A3

[of Sreenivasaprasad & Talhinhas 2005]”), for which they proposed that the

two varietal names of Marcelino et al. (2008) should be raised to species rank

150 ... Pennycook

as “C. fioriniae” (anamorph) and “G. fioriniae” (teleomorph). Unfortunately,

both of these names are invalid: the proposal of “Colletotrichum fioriniae”

cites (with a full and direct reference) the invalid name “C. acutatum var.

fioriniae” as its “basionym’, and fails to supply the required Latin diagnosis

or description; the proposal of “Glomerella fioriniae” fails to provide the

required full and direct reference to its basionym (McNeill et al. 2012: Art

41.5) and indeed does not clearly indicate the name of its basionym (although

in subsequent discussion, G. acutata var. fioriniae is mentioned in passing).

This leaves Glomerella acutata var. fioriniae as the only valid name available

for the taxon.

The species has been widely treated under the invalid name “Colletotrichum

fioriniae” in recent taxonomic and phytopathological literature (e.g., Baroncelli

et al. 2015; Braganga et al. 2016; Damm et al. 2012; Garibaldi et al. 2016; Hyde

et al. 2009; Ivic et al. 2013; Kasson et al. 2014; Kou et al. 2014; Liu et al. 2016;

Munda 2014, 2016; Oo et al. 2016; Pszczdtkowska et al. 2016; Sun et al. 2012;

Zhu et al. 2015). A valid Colletotrichum name is an urgent necessity, because

Colletotrichum 1831, rather than Glomerella 1903, has been selected as the

preferred holomorphic genus name (Réblova et al. 2016: 135).

This unsatisfactory situation is easily remedied under the current

International Code of Nomenclature for algae, fungi, and plants (ICN

(Melbourne); McNeill et al. 2012: Art. 59), in which the separation between

“anamorph” and “teleomorph” names has been abolished. Here, I propose the

name Colletotrichum fioriniae, with basionym Glomerella acutata var. fioriniae.

Taxonomy

Colletotrichum fioriniae (Marcelino & Gouli) Pennycook, comb. & stat. nov.

INDEXFUNGORUM IF 553097

= Glomerella acutata var. fioriniae Marcelino & Gouli, in Marcelino et al.,

Mycologia 100: 362. 2008.

Ho.otype: USA, New York, Westchester, Ward Pound Ridge Reserve, from

mummified adult Fiorinia externa Ferris (Insecta, Diaspididae), “liquid-preserved

perithecia (90% ethanol) produced by crossing Colletotrichum acutatum var.

fiorinia [sic] strains EHS,, x EHS,, as well as mature mycelium of strain EHS,,

in PDA agar cubes’, 2005, J. Marcelino & S. Gouli (Worldwide Collection of

Entomopathogenic Fungi, Entomology Research Laboratory, University of

Vermont, Burlington VT, USA; ex-type culture, CBS 128517 = ARSEF 10222).

“Colletotrichum acutatum var. fioriniae” Marcelino & Gouli, Mycologia 100: 365. 2008,

nom. inval. (no Latin diagnosis or description; ICN (Melbourne) Art. 39.1).

“Colletotrichum fioriniae” R.G. Shivas & Y.P. Tan, Fungal Diversity 39: 117. 2009,

nom. inval. (no Latin diagnosis or description; ICN (Melbourne) Art. 39.1).

“Glomerella fioriniae” R.G. Shivas & Y.P. Tan, Fungal Diversity 39: 117. 2009, nom.

inval. (no full and direct reference to basionym; ICN (Melbourne) Art. 41.5).

Colletotrichum fioriniae comb. & stat. nov. ... 151

Discussion

The validation of Colletotrichum fioriniae as a comb. & stat. nov. has

been preferred because it recognizes the existence of the valid basionym in

Glomerella. Alternatively, the name could have been validated as a sp. nov.,

since the varietal name in Glomerella does not have priority at specific rank.

Acknowledgments

I wish to thank Paul Kirk and Tom May for helpful discussions and for

pre-submission refereeing, and Bevan Weir for advice on the International

Subcommission on Colletotrichum Taxonomy’s acceptance of Colletotrichum as the

preferred generic name.

Literature cited

Baroncelli R, Zapparata A, Sarrocco S, Sukno SA, Lane CR, Thon MR, Vannacci G, Holub E,

Sreenivasaprasad S. 2015. Molecular diversity of anthracnose pathogen populations associated

with UK strawberry production suggests multiple introductions of three different Colletotrichum

species. PLoS ONE 10(6): e0129140. https://doi.org/10.1371/journal.pone.0129140

Braganca CAD, Damm U, Barocelli R, Massola Junior NS, Crous PW. 2016. Species of the

Colletotrichum acutatum complex associated with anthracnose diseases of fruit in Brazil.

Fungal Biology 120(4): 547-561. https://doi.org/10.1016/j.funbio.2016.01.011

Damm U, Cannon PF, Woudenberg JHC, Crous PW. 2012. The Colletotrichum acutatum species

complex. Studies in Mycology 73: 37-113. https://doi.org/10.3114/sim0010

Garibaldi A, Gilardi G, Franco-Ortega S, Gullino ML. 2016. First report of leaf spot caused by

Colletotrichum fioriniae on Mexican bush sage (Salvia leucantha) in Italy. Plant Disease 100(3):

654. https://doi.org/10.1094/PDIS-09-15-1001-PDN

Hyde KD, Cai L, Cannon PF, Crouch JA, Crous PW, Damm U, Goodwin PH, Chen H, Johnston

PR, Jones EBG, Liu ZY, McKenzie EHC, Moriwaki J, Noireung P, Pennycook SR, Pfenning

LH, Prihastuti H, Sato T, Shivas RG, Tan YP, Taylor PWJ, Weir BS, Yang YL, Zhang JZ. 2009.

Colletotrichum - names in current use. Fungal Diversity 39: 147-182.

Ivic D, Voncina D, Sever Z, Simon S, Pejic I. 2013. Identification of Colletotrichum species

causing bitter rot of apple and pear in Croatia. Journal of Phytopathology 161(4): 284-286.

https://doi.org/10.1111/jph.12039

Kasson MT, Pollok JR, Benhase EB, Jelesko JG. 2014. First report of seedling blight of eastern

poison ivy (Toxicodendron radicans) by Colletotrichum fioriniae in Virginia. Plant Disease

98(7): 995. https://doi.org/10.1094/PDIS-09-13-0946-PDN

Kou LP, Gaskins V, Luo YG, Jurick WM. 2014. First report of Colletotrichum fioriniae causing

postharvest decay on ‘Nittany’ apple fruit in the United States. Plant Disease 98(7): 993.

https://doi.org/10.1094/PDIS-08-13-0816-PDN

Lardner R, Johnston PR, Plummer KM, Pearson MN. 1999. Morphological and molecular

analysis of Colletotrichum acutatum sensu lato. Mycological Research 103: 275-285.

https://doi.org/10.1017/S0953756298007023

Liu LP, Yang LY, Liu YN, Yang LN, Lu BH, Yu L, Jin XS, Wang X, Yang C, Li Y, Gao J, Hsiang T. 2016.

First report of anthracnose disease caused by Colletotrichum fioriniae on barbary wolfberry in

China. Plant Disease 100(12):, 2534. https://doi.org/10.1094/PDIS-06-16-0930-PDN

Marcelino J, Giordano R, Gouli S, Gouli V, Parker BL, Skinner M, TeBeest D, Cesnik R. 2008.

Colletotrichum acutatum var. fioriniae (teleomorph: Glomerella acutata var. fioriniae var. nov.)

infection of a scale insect. Mycologia 100(3): 353-374. https://doi.org/10.3852/07-174R

152 ... Pennycook

McNeill J, Barrie FR, Burdet HM, Demoulin V, Hawksworth DL, Marhold K, Nicolson DH,

Prado J, Silva PC, Skog JE, Wiersema JH, Turland NJ. 2006. International Code of Botanical

Nomenclature (Vienna Code). Regnum Vegetabile 146.

http://www.iapt-taxon.org/icbn/main.htm

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

Munda A. 2014. First report of Colletotrichum fioriniae and C. godetiae causing apple bitter rot in

Slovenia. Plant Disease 98(9): 1282. https://doi.org/10.1094/PDIS-04-14-0419-PDN

Munda A. 2016. Outbreak of anthracnose on apricots caused by Colletotrichum fioriniae in

Slovenia. New Disease Reports 33, 2. https://doi.org/10.5197/j.2044-0588.2016.033.002

Oo MM, Tweneboah S, Oh SK. 2016. First report of anthracnose caused by Colletotrichum

fioriniae on Chinese matrimony vine in Korea. Mycobiology 44(4): 325-329.

https://doi.org/10.5941/MYCO.2016.44.4.325

Pszczotkowska A, Okorski A, Paukszto L, Jastrzebski J. 2016. First report of anthracnose disease

caused by Colletotrichum fioriniae on blueberry in western Poland Plant Disease 100(10):

2167. https://doi.org/10.1094/PDIS-04-16-0425-PDN

Réblova M, Miller AN, Rossman AY, Seifert KA, Crous PW, Hawksworth DL, Abdel-Wahab

MA, Cannon PF, Daranagama DA, De Beer ZW, Huang SK, Hyde KD, Jayawardena R,

Jaklitsch W, Jones EBG, Ju YM, Judith C, Maharachchikumbura SSN, Pang KA, Petrini

LE, Raja HA, Romero AI, Shearer C, Senanayake IC, Voglmayr H, Weir BS, Wijayawarden

NN. 2016. Recommendations for competing sexual-asexually typified generic names in

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131-153. https://doi.org/10.5598/imafungus.2016.07.01.08

Shivas RG, Tan YP. 2009. A taxonomic reassessment of Colletotrichum acutatum, introducing

C. fioriniae comb. et stat. nov. and C. simmondsii sp. nov. Fungal Diversity 39: 111-122.

Sreenivasaprasad S, Talhinhas P. 2005. Genotypic and phenotypic diversity in Colletotrichum

acutatum, a cosmopolitan pathogen causing anthracnose on a wide range of hosts. Molecular

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Sun W, Su YY, Cai L, Sun W, Sha W. 2012. First report of leaf disease on Cinnamomum

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MY COTAXON

January-March 2017— Volume 132, pp. 153-175

http://dx.doi.org/10.5248/132.153

Records of terricolous lichens

from paramos of southern Ecuador

YADIRA GONZALEZ’, GREGORIO ARAGON?,

ANA ROSA BURGAZ? & MARIA PRIETO?

"Departamento de Ciencias Biolégicas, Universidad Técnica Particular de Loja,

Loja, Apartado postal 11-01-618, Ecuador

Departamento de Biologia y Geologia, Area de Biodiversidad y Conservacion,

Universidad Rey Juan Carlos, Mostoles, 28933, Spain

*Departamento de Biologia Vegetal II, Facultad de Farmacia, Universidad Complutense,

Madrid, 28040, Spain

CORRESPONDENCE TO: xygonzalez@utpl.edu.ec

ABSTRACT—Ecological studies of five paramos in Azuay and Loja provinces recorded one

lichen species new to South America (Bryoria nitidula), five new to Ecuador (Cladonia

halei, C. melanopoda, C. merochlorophaea, C. subreticulata, Diploschistes diacapsis), two new

to mainland Ecuador (Cladonia grayi, C. pyxidata), and 20 new provincial records. Brief

morphological descriptions, with remarks on distribution and ecology, are provided.

Key worps—Andes, biodiversity, Cladoniaceae, Neotropics

Introduction

The paramo is a Neotropical ecosystem located between the upper forest

line and the permanent snow line. In southern Ecuador these ecosystems are

found from 2800 m altitude, due to local factors such as climate, soil, and the

degree of human intervention (Leon-Yanez 2000, Medina & Mena 2001).

The paramo flora is extremely diverse and considered the richest high

mountain flora of the world (Smith & Cleef 1988, Luteyn 1999). Unfortunately,

its lichen communities are relatively poorly studied (Sipman 2002a). Within

the relevant published information we should emphasize the studies by Sipman

(1992, 1999, 2005), which reported a total of 463 lichen species for Neotropical

paramos (264 species from Colombia, 204 from Costa Rica, and 191 from

154 ... Gonzalez & al.

Ecuador). Also, in monographs on Cladoniaceae (Ahti 1992, 2000), 50 species

of this family were reported from the Andean paramos and subparamos from

Costa Rica to Bolivia, 39 of them above 2800 m in paramo ecosystems in

Ecuador.

Despite these previous studies, the lichen biota of Neotropical paramos

still deserves further study, since the detailed distributions of these taxa are

not well known (Ahti 1992, Sipman 2002a) and lichen flora in Ecuador has

been poorly studied in general (Liicking 1999, Noske & Sipman 2004, Paredes

2006). Our ongoing studies of the Ecuadorian lichen biota have compiled ca.

900 lichen species for mainland Ecuador (Cevallos 2012); comparison with the

ca. 800 species reported from the Galapagos Islands (Bungartz et al. 2013) and

the estimate of 2700 species for Ecuador (Ltcking et al. 2009) underscores the

need for further research on lichen flora in Ecuador.

Thus our goal is to contribute to the knowledge of the lichen flora in paramo

ecosystems and in Ecuador overall.

Material & methods

The specimens were collected in five paramos in Loja and Azuay provinces,

southern Ecuador: Cajanuma, El Cajas, Jimbura, Loma del Oro, and Punzara,

located at altitudes between 2770 and 4040 m. The samples were collected

during ecological studies carried out in 2010 and 2014 (Gonzalez et al. 2017).

Dried specimens were morphologically and anatomically examined using a

stereomicroscope and compound microscope. For species identification we

followed Lumbsch (1989), Ahti (2000), Brodo et al. (2001), Smith et al. (2009),

and Rivas Plata et al. (2010). Secondary compounds were detected by spot tests

and thin-layer chromatography (TLC). Specimen dimensions and chemotype

nomenclature follow Ahti (2000). The specimens are stored in the Herbarium

of Universidad Técnica Particular de Loja, Loja, Ecuador (H-UTPL).

Results

We identified one species new to South America (marked ***), five new to

Ecuador (marked **), two new to mainland Ecuador (marked *), and 20 new

provincial records.

Alectoria ochroleuca (Hoffm.) A. Massal., Sched. Critic. 2: 47. 1855.

Spot test: cortex KC+ yellow, PD-; medulla KC+ yellow or KC-, CK+

yellow-gold, PD-.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46’53”S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M.

Prieto (H-UTPL YG-0060); 2°46’47”S 79°12’20”W, 3788 m, 13.3.2014, A. Benitez & Y.

Terricolous lichens from Ecuador ... 155

Gonzalez (H-UTPL YG-0303); 2°47’39”S 79°12’09” W, 3894 m, 26.3.2014 (H-UTPL YG-

0304); 2°47’03”S 79°13’30”W, 4009 m, 15.2.2014 (H-UTPL YG-0305).

Alectoria ochroleuca is characterized by its erect to decumbent shrubby thallus

with greenish yellow branches darkening towards the tips (Brodo et al. 2001).

Widespread in Asia, Central and North America, Europe, and Oceania

(Guzman-Davalos & Alvarez 1987, Smith et al. 2009), in South America

A. ochroleuca is distributed in Argentina, Bolivia, Chile, Colombia, Costa Rica,

Ecuador, Peru, and Venezuela (Nylander 1861, Galloway & Quilhot 1998,

Calvelo & Liberatore 2002, Umafa-Tenorio et al. 2002, Sipman 1999, Ramos

2014).

This species grows on the ground or rarely on shrubs and preferably in

arctic-alpine ecosystems (Brodo et al. 2001). In Ecuador, it was previously

cited in Chimborazo, Cotopaxi, Loja, and Pichincha provinces (Zahlbruckner

1905, 1907; Arvidsson 1991; Sklenaf et al. 2010; Benitez et al. 2012). Here it is

reported for the first time from Azuay province (in El Cajas paramo).

Arthrorhaphis alpina (Schaer.) R. Sant., Lichenologist 12: 106. 1980.

SPECIMENS EXAMINED—ECUADOR, Loja. Loja: Punzara, 4°02’36”’S 79°13’59’W,

2770 m, paramo, 19.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0105);

Saraguro: Loma del Oro, 3°40’52”S 79°14’24’W, 3245 m, paramo, 19.8.2010, A. Benitez,

Y. Gonzalez & M. Prieto (H-UTPL YG-0106).

The thallus of A. alpina is composed of more or less rounded, convex, bullate

squamules, <1 mm diam., yellow to yellowish green, matte, and with a more or

less pruinose roughened surface. Apothecia are infrequent, located between the

squamules, blackish grey-green.

Arthrorhaphis alpina is widespread in Asia, Europe, North America,

and Oceania (Obermayer 1994, Esslinger & Egan 1995, Elix & McCarthy

1998, Aptroot & Feijen 2002, Aptroot & Sparrius 2003). In South America is

distributed in Bolivia, Colombia, Chile, Costa Rica, Ecuador, and Venezuela

(Hafellner & Obermayer 1995, Sipman 1999, Galloway & Quilhot 1998). The

species grows on soil and among mosses (Smith et al. 2009).

Previously found in Ecuador in Pichincha province (Arvidsson 1991), this is

the first record of A. alpina for Loja province.

Bryoria nitidula*** (Th. Fr.) Brodo & D. Hawksw., Op. Bot. 42: 107. 1977. FIG. 1

Fruticose, shrubby thallus, 4-8 cm, erect or spreading horizontally over the

ground, branching anisotomic and usually dichotomous towards the apices.

Branches dark brown to black, darker at the base, shiny and terete, 0.5-0.9 mm

diam., with a few lateral perforations or fissures of 0.4-0.8 mm; lacking soredia

or isidia.

156 ... Gonzalez & al.

Figure 1. Bryoria nitidula.

a. Thallus. b. Branching of podetia. Scale bars: a= 5 mm; b = 2 mm.

Substances detected by TLC: none.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46’53”S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M.

Prieto (H-UTPL YG-0057); 2°47’39”S 79°12’09’W, 3890 m, 26.3.2014, A. Benitez & Y.

Gonzalez (H-UTPL YG-00301); 2°46’47”S 79°12’21”W, 3789 m, 13.3.2014 (H-UTPL

YG-00298).

Bryoria nitidula occurs in Asia, Europe, and North America, growing on arctic

and subarctic heaths (Smith et al. 2009). It was found for the first time in

Ecuador being locally common in El Cajas paramo, at 3750-4020 m elevations.

This is the first record in South America.

Our specimens closely resemble those described from North America, but

the Ecuadorian samples do not contain fumarprotocetraric acid.

Cladia aggregata (Sw.) Nyl., Bull. Soc. Linn. Normandie, Ser. 2, 4: 167. 1870.

Substances detected by TLC: barbatic and 4-O-demethylbarbatic acids

(chemotype I).

SPECIMENS EXAMINED—ECUADOR, Loja. Loja: Parque Nacional Podocarpus,

Cajanuma, 4°06’59”S 79°09’41”W, 3337 m, paramo, 29.6.2010, A. Benitez, Y. Gonzalez

& M. Prieto (H-UTPL YG-0064); 13.7.2010 (H-UTPL YG-0079); Punzara, 4°02’36”S

79°13'59’'W, 2770 m, paramo, 19.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0100); Saraguro: Loma del Oro, 3°40’52”S 79°14’24’W, 3245 m, paramo, 19.8.2010,

A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0005 and H-UTPL YG-0006);

Espindola: Jimbura, 4°42’37”S 79°25’46”W, 3450 m, paramo, 8.7.2010, A. Benitez, Y.

Gonzalez & M. Prieto (H-UTPL YG-0082 and H-UTPL YG-0083).

The thallus of C. aggregata is characterized by its dark brown to blackish, pale

yellowish to straw-colored or pale green pseudopodetia, 5-150 x 0.5-8 mm.

Terricolous lichens from Ecuador ... 157

The pseudopodetial wall is cartilaginous with infrequent to abundant rounded

to elliptic perforations; the central canal surface is white and loosely arachnoid

(Ahti 2000).

Although several species have been described with the same chemosyndrome

within the Cladia aggregata complex (Parnmen et al. 2012, 2013), their

distribution and morphology do not match our samples. Moreover, the

barcoding molecular sequences produced for our samples (nuITS region) place

our samples with Cladia aggregata s. str.

Cladia aggregata s. lat. is distributed in the Southern Hemisphere, including

Australasia, Southeast Asia to South Japan and India, and South Africa (Ahti

2000). In the Neotropical region it is widespread, from Mexico and the West

Indies to the subantarctic islands, but it is absent in lowland and arid regions

(Ahti 2000). It is very common in montane areas, on soil, and wood (Ahti

2000). In Ecuador four chemotypes (II, IH, IV, and VII) have previously been

reported (Ahti 2000). Chemotype I was found for the first time in mainland

Ecuador growing in paramos of Azuay and Loja provinces. This chemotype

was previously reported in Galapagos Islands (Yanez-Ayabaca et al. 2013). Our

specimens constitute the first records from Loja province.

Cladia fuliginosa Filson, Victorian Nat. 87: 325. 1970

Substances detected by TLC: divaricatic and usnic acids.

SPECIMENS EXAMINED—ECUADOR, Loja. Loja: Parque Nacional Podocarpus,

Cajanuma, 4°06’59”S 79°09’41” W, 3337 m, paramo, 29.6.2010, A. Benitez, Y. Gonzalez

& M. Prieto (H-UTPL YG-0081); Punzara, 4°02’36”S 79°13’59”W, 2770 m, paramo,

19.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0098 and H-UTPL YG-

0099); Saraguro: Loma del Oro, 3°40’52”S 79°14’24”W, 3245 m, paramo, 7.7.2010,

A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0001 and H-UTPL YG-0002);

Espindola: Jimbura, 4°42’37”S 79°25’46”W, 3450 m, paramo, 8.7.2010, A. Benitez,

Y. Gonzalez & M. Prieto (H-UTPL YG-0091).

Cladia fuliginosa produces green to blackish brown pseudopodetia, <70 x 2 mm;

the perforated and irregularly branched pseudopodetia are mostly filled by a

dark brown to black inner medulla (Ahti 2000).

Distributed throughout the Andes, from Colombia to Peru growing on

soil in paramos, C. fuliginosa is also present in Australia and New Zealand

(Ahti 2000). In Ecuador, it has previously been cited in Azuay, Imbabura, and

Pichincha provinces (Arvidsson 1991, Ahti 2000). Our collections constitute

the first records of C. fuliginosa from Loja province.

Cladonia aleuropoda Vain., Beibl. Hedwigia 38: 190. 1899.

Substances detected by TLC: fumarprotocetraric and often quaesitic acids

(chemotype I) and grayanic acid (chemotype II).

158 ... Gonzalez & al.

SPECIMEN EXAMINED—ECUADOR, Loja. Loja: Punzara, 4°02’36”S 79°13’59”W, 2770

m, paramo, 19.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPLYG-0104).

The thallus produces 3-10 cm tall unbranched or sparsely branched whitish to

brownish gray podetia. Initially subulate, the podetial tips later produce 1-7

mm wide scyphi (Ahti 2000).

Cladonia aleuropoda is distributed at 2200-4500 m elevations along the

Andes in Colombia, Costa Rica, Ecuador, Guatemala, México, Peru, and

Venezuela, growing on soil and rocks in paramos and upper cloud forests (Ahti

2000). Chemotypes I and II have been previously recorded in Ecuador from

Azuay, Carchi, Chimborazo, Cotopaxi, Napo, and Pichincha provinces (Ahti

2000). This is the first record of chemotype I for Loja province.

Cladonia arbuscula (Wallr.) Flot., Thermen Warmbr. Riesengeb.: 94. 1839, s.lat.

Substances detected by TLC: fumarprotocetraric and usnic acids

(chemotype I).

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53’S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M. Prieto

(H-UTPL YG-0041); 2°4660’S 79°13’61”W, 4012 m, 22.2.2014 (H-UTPL YG-0275);

2°47’40”S 79°12’08”W, 3893 m (H-UTPL YG-0278); 2°47’39”S 79°12’09”W, 3891 m,

26.3.2014 (H-UTPL YG-0274); 2°46’47S 79°12’19” W, 3789 m, 14.3.2014 (H-UTPL YG-

0276); 2°46’50”S 79°12’20’W, 3787 m, 13.3.2014 (H-UTPL YG-0273). Loja. Saraguro:

Loma del Oro, 3°40’52”S 79°14’24”W, 3245 m, paramo, 19.8.2010, A. Benitez, Y.

Gonzalez & M. Prieto (H-UTPL YG-0037); Espindola: Jimbura, 4°42’37”S 79°25’46”W,

3450 m, paramo, 8.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0092).

Under C. arbuscula, we include specimens representing the type subspecies

and the subspecies boliviana. Piercey-Normore et al. (2010), who demonstrated

that molecular data do not support the existence of some subspecies, did not

include subsp. boliviana, and so additional molecular analyses are needed to

test the status of subsp. boliviana.

C. arbuscula subsp. boliviana (Ahti) Ahti & DePriest is characterized by

yellowish to whitish gray podetia with slightly brownish tips. It grows up to

17 cm, its podetia are slender, generally loosely branched, with its ultimate

branchlets erect to divaricate, rather dense at apex but rarely strongly unilaterally

deflexed (Ahti 2000). The subspecies differs from C. subsp. arbuscula mainly

in its darker color, bigger size, and highly frequent dichotomous branching,

and the absence of psoromic acid (Ahti 2000). However, these characters could

be misleading and molecular data are necessary for the identification of the

Andean material (Ahti pers. comm.).

Cladonia arbuscula is common in paramos and upper Andean forests (Ahti

2000) in Bolivia (chemotype J), Brazil (I, II), Colombia (I, IL, II), and Venezuela

(I, IL, III) but rare in Costa Rica (II) and Guatemala (I). In Ecuador chemotype

Terricolous lichens from Ecuador ... 159

II (with usnic acid only) has been previously found in the provinces of Azuay,

Galapagos Islands, Imbabura, Loja, and Tungurahua (Ahti 2000). Chemotype

I was found for the first time in Ecuador, growing in three different paramos.

Cladonia coccifera (L.) Willd., Fl. Berol. Prodr.: 361. 1787.

Substances detected by TLC: zeorin and usnic acid.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53’S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M. Prieto

(H-UTPL YG-0052 and H-UTPL YG-0053); 2°46’46”S 79°12’20”W, 3786 m, 13.3.2014,

A. Benitez & Y. Gonzalez (H-UTPL YG-0160).

Cladonia coccifera is characterized by its persistent primary thallus, consisting

of ascending squamules, often with orange basal parts. The 0.7-3.5 cm tall

podetia are greenish, yellowish or whitish gray, with an areolate-corticate

surface and totally covered by pruinose granules, schizidia, and phyllidia

(also inside the scyphi). Red-coloured hymenial discs are fairly common

(Ahti 2000).

Distributed in the Northern Hemisphere in Himalaya, Java, and Papua New

Guinea, C. coccifera has been found at elevations of 2400-4300 m throughout

Central and South America in Argentina, Bolivia, Brazil, Colombia, Costa Rica,

Ecuador, Guatemala, Hispaniola, Venezuela, and Peru (Sipman 1999, Ahti

2000, Calvelo & Liberatore 2002, Flakus et al. 2008), growing on humous soils

and occasionally on wood and plant debris. Previously reported for Ecuador in

Carchi, Cotopaxi, Loja, and Zamora-Chinchipe (Ahti 2000, Noske & Sipman

2004, Mandl 2007), this is the first record of C. coccifera for Azuay province.

Cladonia corniculata Ahti & Kashiw., Stud. Cryptog. South. Chile: 136. 1984.

Substances detected by TLC: fumarprotocetraric acid.

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’W, 3245 m, paramo, 19.8.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0028); Loja: Parque Nacional Podocarpus, Cajanuma, 4°06’59”S 79°09’41”W, 3337

m, paramo, 13.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0067, H-UTPL

YG-0068, H-UTPL YG-0069 and H-UTPL YG-0080).

Cladonia corniculata is characterized by 12-60 x 0.5-2.5 mm white, green,

or whitish gray podetia. Podetia are flexuous or not, at first unbranched and

later dichotomously branched, with subulate tips, always ascyphose, sometimes

with obtuse or dilated branchlets. Thallus surface is completely ecorticate with

farinose to slightly granulose soredia (Ahti 2000).

Distributed throughout Asia, Africa, Australia, New Zealand, and the

subantarctic islands, C. corniculata is widespread in Central and South America

(especially at high elevations in the Andes). It grows on moist and shady road

banks in paramos being rare on bare soils (Stenroos et al. 1992, Ahti 2000).

160 ... Gonzalez & al.

In Ecuador the species was previously reported from Azuay, Carchi,

Cotopaxi, Galapagos Islands, Imbabura, Napo, and Pichincha (Ahti 2000); our

collections represent the first records for Loja province.

Cladonia crispata (Ach.) Flot., Thermen Warmbr. Riesengeb.: 93. 1839.

Substances detected by TLC: squamatic acid.

SPECIMEN EXAMINED—ECUADOR, Loja. Loja: Punzara, 4°02’36”S 79°13’59’W, 2770

m, paramo, 19.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0097).

The thallus of C. crispata is characterized by its 20-80 x 0.5-2 mm brownish

gray podetia that are not blackish at the base. The podetia have dilated apices,

1-3 mm wide axillary funnels, and a smooth surface (Ahti 2000).

A subcosmopolitan species, distributed throughout Australasia, East Africa,

Asia, Europe, New Guinea, and North America (Ahti 2000, Burgaz & Ahti

2009), C. crispata has been recorded throughout Central and South America

in Colombia, Costa Rica, Dominican Republic, Ecuador, Jamaica, Peru, and

Venezuela (Ahti 2000) as locally common in paramos and upper Andean

forests where it grows on soil and wood at 1700-3300 m altitudes (Ahti 2000).

Previously recorded in Ecuador from Zamora-Chinchipe (Néske & Sipman

2004) in upper montane forests, our first collection in paramo in Loja province

widens the distribution and ecology of Cladonia crispata in Ecuador.

Cladonia grayi* G. Merr. ex Sandst., Sandstede Clad. Exs.: no. 1847. 1929. Fi. 2

Podetia 1-2 cm, scyphose, greenish gray, often brownish in basal parts, with

proliferations from scyphal margins, scyphi 2-4.5 mm diam. Surface rough,

verruculose, upper part and interior of scyphi granulose, clearly sorediate or

eroded in some parts, with age developing macrosquamules. Hymenial discs

frequent, dark brown.

Substances detected by TLC: fumarprotocetraric and grayanic acids.

SPECIMEN EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'47’S 79°14’57”W, 3789 m, paramo, 13.3.2014, A. Benitez & Y. Gonzalez (H-UTPL

YG-0165).

Cladonia grayi, widespread in the Northern Hemisphere from the Arctic to

temperate regions (where it is very common in eastern North America) and is

also present in Australasia (Ahti 2000, Burgaz & Ahti 2009). In the Neotropical

region it is scattered throughout Argentina, Bolivia, Brazil, Colombia, Costa

Rica, Cuba, Dominican Republic, El Salvador, Galapagos Islands, Guatemala,

Haiti, Honduras, Jamaica, Paraguay, Uruguay, and Venezuela, growing at high

elevations on highly acidic soils (Ahti 2000, Burgaz & Ahti 2009, Yanez-Ayabaca

et al. 2013). Our collection from one locality in Azuay province represents a

first record of C. grayi from mainland Ecuador.

Terricolous lichens from Ecuador... 161

FiGuRE 2. Cladonia grayi. a. Macrosquamules on margin of scyphi.

b. Granules on upper part of scyphi. Scale bars: a = 3 mm; b = 2 mm.

Although similar to specimens described for the Neotropical and

temperate regions (Ahti 2000), our Ecuadorian material is does not develop

macrosquamules at the base and is found at higher altitudes (3789 m).

Cladonia halei** (Ahti) Ahti & DePriest, Mycotaxon 78: 501. 2001. Fic. 3

Podetia ash gray or brownish gray, darkening when exposed. ‘Thallus

<12 cm tall, with anisotomic branches, usually deflexed, with clearly distinct

0.6-2 mm thick main axis. Podetia surface markedly verruculose with

arachnoid interspaces.

Substances detected by TLC: fumarprotocetraric acid.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53’S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M. Prieto

(H-UTPL YG-0050 and H-UTPL YG-0051); 2°47’08”S 79°13’32”W, 4032 m, 8.2.2014,

A. Benitez & Y. Gonzalez (H-UTPL YG-0299); 2°47’38’S 79°12’09” W, 3884 m, 25.3.2014

(H-UTPL YG-0286); 2°46’46”S 79°12’20”W, 3788 m, 13.3.2014 (H-UTPL YG-0300).

Cladonia halei has been previously recorded in Colombia, Papua New Guinea,

Peru, and Venezuela (Ahti 2000 and pers. comm.), growing in paramo and

subparamo zones at 2500-4000 m altitudes in wet Sphagnum bogs and on drier

ground in humid montane regions (Ahti 2000). Our discovery of C. halei for

the first time in Ecuador growing in El Cajas paramo considerably widens its

world distribution.

162 ... Gonzalez & al.

Ficure 3. Cladonia halei.

a. Thallus. b. Branching of podetia. Scale bars = 5 mm.

Our specimens are similar in morphology, chemistry, and ecology to that

described for the Neotropical region.

Cladonia leprocephala Ahti & S. Stenroos, Ann. Bot. Fenn. 23: 236. 1986.

Substances detected by TLC: thamnolic acid.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53”S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M.

Prieto (H-UTPL YG-0046); 2°47’37”S 79°12’09”W, 3891 m, 25.3.2014, A. Benitez &

Y. Gonzalez (H-UTPL YG-0166).

Cladonia leprocephala is characterized by its 8-23 x 0.8-2 mm greenish to

whitish gray podetia with scyphi having subentire margins 2-3 mm wide.

Thallus surface in generally basally areolate-corticate up to the scyphi, with

granulose soredia. The red hymenial discs are rare and present at the tips of

proliferations from scyphal margins (Ahti 2000).

Distributed along the Andes in Colombia, Costa Rica, Ecuador, Peru, and

Venezuela in paramos at 2900-4250 m elevations, growing on moist and

humous soils, on wood, and as epiphyte on shrubs at timberline (Ahti 2000,

pers. comm.), in Ecuador C. leprocephala has been previously reported from

Cotopaxi, Napo, Pichincha, and Tungurahua (Ahti 2000). Our collections

represent the first records of the species for Azuay province.

Terricolous lichens from Ecuador ... 163

Cladonia lopezii S. Stenroos, Ann. Bot. Fenn. 26: 250. 1989.

Substances detected by TLC: didymic acid.

SPECIMEN EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53”S 79°13’21” W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M.

Prieto (H-UTPL YG-0106).

The species has <25 x 30 mm brown podetia that are marginal on primary

squamules, which elongate and roll up to form the podetial tube, abruptly

flaring to form conspicuously dorsiventrally flattened web-like structures.

The podetia usually produce red hymenial discs, forming agglomerations up

to 9 mm wide (Ahti 2000).

Cladonia lopezii occurs in the high Andes from Bolivia to Venezuela

and in Brazil at 1200-3935 m altitudes, growing on soil, rocks, and rotten

wood (Ahti 2000). The species has been previously found in Loja, Morona-

Santiago, and Zamora-Chinchipe (Ahti 2000, Noske & Sipman 2004, Mandl

2007); our collection constitutes the first record for Azuay province.

Cladonia macilenta Hoftm., Deutschl. Fl. 2: 126. 1796.

Substances detected by TLC: barbatic acid [chemotype IJ].

SPECIMEN EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'46"S_ 79°12’18”W, 3793 m, paramo, 14.3.2014, A. Benitez & Y. Gonzalez

(H-UTPL YG-0199).

The thallus of C. macilenta is characterized its 5-40 x 0.3-2.5 mm whitish

ascyphose podetia with subulate to blunt apices that are swollen below the

hymenia, unbranched or sometimes branched (Ahti 2000, Burgaz & Ahti

2009). The thallus surface is mostly covered by abundant farinose-soredia

(Ahti 2000, Burgaz & Ahti 2009).

Substances detected by TLC: barbatic acid. There are two major

chemotypes recognized, chemotype I with thamnolic acid as major

compound, and chemotype II with barbatic acid as major compound.

Rare in tropical lowlands and highest mountains, C. macilenta is

widespread between 1500-3250 m along the northern Andes, growing on

wood, stumps, tree bases, peat bogs, acid rocks, and humous or sandy acid

soils (Ahti 2000, Burgaz & Ahti 2009). In Ecuador it has been previously

found in Carchi (chemotype II), Galapagos Islands (chemotypes I and II),

and Tungurahua (chemotype II) (Ahti 2000, Yanez-Ayabaca et al. 2013).

Our specimen (chemotype IJ) represents a first record for Azuay province.

164 ... Gonzalez & al.

Figure 4. a. Cladonia melanopoda. Small squamules on surface of podetium. Scale bar = 2 mm.

b. Cladonia merochlorophaea. Phyllidia and schizidia inside scyphi. Scale bar = 2 mm.

Cladonia melanopoda** Ahti, Symb. Bot. Upsal. 32(1): 7. 1997. Fic. 4a

Podetia 45-60 x 0.5-1 mm, slender, unbranched to slightly branched,

brownish to blackish at the base. The podetial surface either totally decorticated

or with areolate schizidia or small squamules. Scyphi 1-4 mm wide, always

with central proliferations in 5-7 tiers.

Substances detected by TLC: fumarprotocetraric acid and traces of quaesitic

acid.

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’'W, 3245 m, paramo, 19.8.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0019, H-UTPL YG-0020, H-UTPL YG-0021).

Previously found at high elevations of the Andean range in Argentina, Bolivia,

and Peru on soil in paramo and timberline vegetation at 1300-4000 m (Ahti

2000), C. melanopoda is reported here for the first time in Ecuador from one

paramo in Loja province.

Our specimens are very close to those described from the Neotropical

region.

Cladonia merochlorophaea** Asahina, J. Jap. Bot. 16: 713. 1940. Fic. 4b

Podetia 10-30 x 1-2 mm, scyphose with 2-6 mm wide scyphi, sometimes

simple or frequently with proliferations from margins and the old bases

strongly blackish. The surface is usually verruculose with wide bare spaces

Terricolous lichens from Ecuador ... 165

and macrosquamules. Inside scyphi there are phyllidia and schizidia often

brownish. The scyphal margin may produce pyriform pycnidia.

Substances detected by TLC: merochlorophaeic and fumarprotocetraric

acids.

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46’60’S 79°13'31”W, 4007 m, paramo, 22.2.2014, A. Benitez & Y. Gonzalez (H-UTPL

YG-0151); 2°47’38”S 79°12’10”W, 3895 m, 25.3.2014 (H-UTPL YG-0155); 2°47’36”S

79°12’10’W, 3895 m (H-UTPL YG-0158); 2°46’47”S 79°12’21”W, 3789 m, 13.3.2014

(H-UTPL YG-0162); 2°46’46”S 79°12’19”W, 3790 m, 14.3.2014 (H-UTPL YG-0290).

A subcosmopolitan species found from arctic to temperate regions in

Antarctic, Australasia, Eurasia, North America, and southern South America,

C. merochlorophaea is found In the Neotropical region in Bolivia, Brazil, Peru,

and Venezuela (Ahti 2000, pers. comm.) usually growing on acid humus,

peat, and wood (Stenroos et al. 1992, Ahti 2000, Burgaz & Ahti 2009). Our

collections constitute the first reports for the species in Ecuador, where it was

being locally common in Azuay province (in El Cajas paramo).

Our specimens are similar in morphology, chemistry, and ecology to that

described for the Neotropical and temperate regions (Ahti 2000).

Cladonia pyxidata* (L.) Hoffm., Deutschl. Fl. 2: 121. 1796. FIG. 5

Primary thallus persistent, squamules 1-3 x 2-3 mm. Scyphose podetia

4-12 mm tall, greenish gray to dark brown, stalks 1-1.5 mm diam., scyphi

0.8-4.5 mm diam., usually simple or sometimes proliferating from margins.

Podetia surface corticated, often continuous but can be areolate or cracked

and usually decorticated above. Inside of scyphi it presents schizidia, phyllidia,

microsquamules, and flat granules, these latter usually larger.

Substances detected by TLC: fumarprotocetraric and homosekikaic acids

(chemotype II).

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°47'37’S 79°12'09’ W, 3889 m, paramo, 25.3.2014, A. Benitez & Y. Gonzalez (H-UTPL

YG-0168); 2°46'47”S 79°12’21’W, 3791 m, 13.3.2014 (H-UTPL YG-0164 and H-UTPL

YG-0167).

Distributed throughout Africa, Asia, Australia, Europe, North America, and

New Zealand (Smith et al. 2009), in the Neotropical region C. pyxidata has

been previously found in Argentina, Brazil, Colombia, Dominican Republic,

El Salvador, Galapagos Islands, Haiti, Honduras, Guatemala, Jamaica, Peru,

Uruguay, and Venezuela, growing on acid substrates in bare and humous

soils, mossy rocks, and rarely on dead wood (Ahti 2000, pers. comm.,

Burgaz & Ahti 2009, Yanez-Ayabaca et al. 2013). Our collection from Azuay

166 ... Gonzalez & al.

FiGuRE 5. Cladonia pyxidata. a. Phyllidia, schizidia, microsquamules, and flat granules inside

scyphi. b. Decorticate podetia surface. Scale bars = 3 mm.

province (in El Cajas paramo) represents the first record of C. pyxidata for

mainland Ecuador. Chemotype II is fairly rare and has previously been

noted only for Guatemala and Honduras (Ahti 2000).

The Ecuadorian material is similar in morphology and ecology to those

described for the Neotropical region (Ahti 2000, Burgaz & Ahti 2009,

Yanez-Ayabaca et al. 2013) but in Ecuadorian samples the scyphi are wider

(<4 mm).

Cladonia squamosa (Scop.) Hoffm., Deutschl. FI. 2: 125. 1796.

Substances detected by TLC: squamatic acid (chemotype I), thamnolic and

barbatic acids (chemotype II).

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’'W, 3245 m, paramo, 7.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0017); Loja: Parque Nacional Podocarpus, Cajanuma, 4°06’59”S 79°09'41”W,

3337 m, paramo, 13.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0066);

Azuay. Cuenca: Parque Nacional El Cajas, 2°47’38”S 79°12’09”W, 3885 m, paramo,

25.3.2014, A. Benitez & Y. Gonzalez (H-UTPL YG-0193); 2°47’37”S 79°12’09”’W, 3891

m, 26.3.2014 (H-UTPL YG-0194); 2°47’39”S 79°12’09”W, 3894 m, 26.3.2014 (H-UTPL

YG-0195); 2°47’38”S 79°12’09”W, 3886 m, 25.3.2014 (H-UTPL YG-0197); 2°46’47”S

79°12’19”"W, 3782 m, 25.3.2014 (H-UTPL YG-0198); 2°46’37”S 79°12’09”W, 3890 m,

25.3.2014 (H-UTPL YG-0202); 2°47’37”S 79°12’09”W, 3889 m, 25.3.2014 (H-UTPL

YG-0203); 2°47’37”S 79°12’09”W, 3889 m, 25.3.2014 (H-UTPL YG-0204); 2°46’46”S

79°12’20’W, 3733 m, 13.3.2014 (H-UTPL YG-0207); 2°47'08”S 79°13’32”W, 4030 m,

9.2.2014 (H-UTPL YG-0209); 2°47’37”S 79°12’04”’W, 3889 m, 25.3.2014, (H-UTPL

YG-0210); 2°47’00”S 79°13’31”W, 4018 m, 22.2.2014 (H-UTPL YG-0211); 2°47’38”S

79°12’09”W, 3880 m, 25.3.2014 (H-UTPL YG-0212); 2°47’37”S 79°12’10”W, 3886 m,

25.3.2014 (H-UTPL YG-0213); 2°46’47”S 79°12’19”W, 3782 m, 14.3.2014 (H-UTPL

Terricolous lichens from Ecuador ... 167

YG-0214); 2°47’08”S 79°13’32”W, 4029 m, 9.2.2014 (H-UTPL YG-0215); 2°47’03”S

79°13/32”W, 4006 m, 16.2.2014 (H-UTPL YG-0221); 2°47’38”S 79°12’10”’W, 3891 m,

25.3.2014 (H-UTPL YG-0222); 2°47’38’S 79°12’09”W, 3883 m, 25.3.2014 (H-UTPL YG-

0236); 2°46’47”S 79°12’21”W, 3788 m, 13.3.2014 (H-UTPL YG-0258).

In Cladonia squamosa, the podetia are 30-140 x 0.5-5 mm, greenish gray to

strongly brown, not black at the base, unbranched or irregularly branched, with

usually dilated axils and perforated apex (Ahti 2000, Burgaz & Ahti 2009). The

thallus surface is never sorediate, but mostly or totally decorticated and densely

covered with crenulate or laciniate 1-8 mm tall squamules and granules (Ahti

2000).

Widespread in temperate and cooler regions in both Hemispheres, also

along the Andes, from Mexico to Chile, and Argentina at 1000-4500 m

altitudes, C. squamosa is also present in southeast Brazil down to 700 m. It

grows in cloud forests and moist paramos, on rocks and acidic and humous

soils or wood (Ahti 2000, Burgaz & Ahti 2009). In Ecuador, the species has

been previously found in Azuay (chemotype I), Carchi (chemotype I), Pastaza

(chemotype II), and Zamora-Chinchipe (chemotype II) provinces (Arvidsson

1991, Ahti 2000, Noske & Sipman 2004). Our collections represent the first

records of chemotype II specimens from Azuay and Loja provinces and the first

record of chemotype I for Azuay province.

Cladonia subreticulata** Ahti, Acta Bot. Fenn. 10: 168. 1973. FIG. 6a

Podetial surface compact and slightly arachnoid, usually forming well-

developed cushions. Podetia 30-50 x 3.5-4.5 mm, thick, yellow to grayish

yellow, with very irregular branching, open apices and axils, and perforated

podetial wall.

Substances detected by TLC: usnic acid (chemotype II).

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24” W, 3245 m, paramo, 19.8.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0023 and H-UTPL YG-0024).

Previously reported from South America in Brazil, Guyana, Peru, and

Venezuela at high elevations in woodlands and cloud forests on sandy soils and

rock outcrops (Ahti 2000, Ahti & Sipman 2013), C. subreticulata was found for

the first time in Ecuador, where it was very rare found growing on relatively

bare soils in Loma del Oro paramo in Loja province.

Our specimens, which morphologically, ecologically, and chemically

resemble those described for the Neotropic Region (Ahti 2000), differ in having

smaller (<5 cm) podetia.

168 ... Gonzalez & al.

FiGuRE 6. a. Cladonia subreticulata. Branching of podetia. Scale bar = 3 mm.

b. Diploschistes diacapsis. Apothecia. Scale bar: 3 mm.

Dibaeis columbiana (Vain.) Kalb & Gierl, Herzogia 9: 621. 1993.

SPECIMEN EXAMINED—ECUADOR, Loja. Espindola: Jimbura, 4°42’37”S 79°25’46” W,

3450 m, paramo, 8.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0088).

Diabaeis columbiana is characterized by a densely granular crustose thallus

with 0.2 mm wide pale gray granules, 2-11 mm tall podetia with convex pink

apothecia, and lacking schizidia or soredia (Sipman 1997).

A terricolous species found in Bolivia, Colombia, Costa Rica, Ecuador, and

Venezuela (Marcano et al. 1996, Sipman 1999, Flakus & Wilk 2006), in Ecuador

D. columbiana was previously found on subalpine dwarf-forest and paramo in

Zamora-Chinchipe (Néske & Sipman 2004). Our collection is the first record

for Loja province, where it is rare.

Diploschistes diacapsis** (Ach.) Lumbsch, Lichenologist 20: 20. 1988. Fra. 6b

Thallus verrucose and areolate, upper surface whitish to whitish-gray,

slightly to abundantly pruinose; apothecia urceolate, 0.5-2.5(-4) mm diam.

black or pruinose gray; ascospores 4-8 per ascus.

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’W, 3245 m, paramo, 19.8.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0034); Azuay. Cuenca: Parque Nacional El Cajas, 2°46’53”S 79°13’21”W, 3930 m,

paramo, 27.7.2010 (H-UTPL YG-0058); 2°46’46”S 79°12’21”W, 3790 m, 13.3.2014, A.

Benitez & Y. Gonzalez (H-UTPL YG-0302).

Diploschistes diacapsis is identified by the K+ yellow reaction of the thallus

due to the presence of diploschistesic acid as major constituent (Rivas Plata

et al. 2010); the PD-, C+ red and KC+ red chemistry is due to lecanoric and

diploschistesic acids.

Terricolous lichens from Ecuador ... 169

Found in Africa, Asia, Australia, Europe, North, and South America

(Lumbsch 1989), D. diacapsis has been reported from South America from

paramo ecosystems in Chile, Costa Rica, and Peru (Lumbsch 1989, Galloway &

Quilhot 1998, Sipman 1999). Our collections constitute the first records from

Ecuador, where it was found growing in two different paramos in Azuay and

Loja provinces.

Our specimens closely resemble those described in the Holarctic zone

(Lumbsch 1989) except for its larger (<4 mm) apothecia.

Normandina pulchella (Borrer) Nyl., Ann. Sci. Nat., Bot., sér. 4, 15: 382. 1861.

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’W, 3245 m, paramo, 13.8.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0107); Espindola: Jimbura, 4°42’37”S 79°25’46’”W, 3450 m, paramo, 8.7.2010, A.

Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0108).

Normandina pulchella is characterized by its very thin 0.7-2.5 mm tall scattered

to crowded green to slightly bluish green squamules with sharply raised

margins. Soralia are often present on lobe surface and margins. The lower

squamule surfaces are whitish and slightly felted with numerous hyphae (Smith

et al. 2009).

A frequent cosmopolitan species (Smith et al. 2009), in the Neotropical

region N. pulchella is found in Argentina, Bolivia, Brazil, Chile, Colombia,

Costa Rica, Ecuador, Guyana, Peru, Uruguay, and Venezuela (Calvelo &

Liberatore 2002, Flakus & Wilk 2006, Sipman 1999). It grows on mosses, other

lichens, bark, and rocks in habitats with high air humidity (Nash et al. 2004). In

Ecuador the species was previously found in Galapagos Islands and Zamora-

Chinchipe (Arvidsson 1991, Elix et al. 1998, Davey 1999, Noske 2004, Néske

& Sipman 2004, Mandl 2007). Our collections represent the first records from

Loja province, where it was found growing on bryophytes.

Phyllobaeis imbricata (Hook.) Kalb & Gierl, Herzogia 9: 610. 1993.

SPECIMENS EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S

79°14’24’W, 3245 m, paramo, 7.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL

YG-0029, H-UTPL YG-0030 and H-UTPL YG-0031); Loja: Parque Nacional

Podocarpus, Cajanuma, 4°06’59”S 79°09’41”W, 3337 m, paramo, 29.6.2010, A. Benitez,

Y. Gonzalez & M. Prieto (H-UTPL YG-0073 and H-UTPL YG-0074).

Phyllobaeis imbricata is distinguished by its squamulose primary thallus with

long deeply lobed squamules, cylindric podetia, and pink terminal apothecia.

A high-elevation Neotropical species in Bolivia, Brazil, Colombia, Costa

Rica, Ecuador, and Venezuela (Gierl et al. 1993, Sipman 1999, Umafia-Tenorio

et al. 2002, Flakus & Wilk 2006), in Ecuador, P imbricata was previously found

170 ... Gonzalez & al.

in Zamora-Chinchipe on subalpine dwarf-forest and paramo (Noéske & Sipman

2004). Our specimens represent the first records for Loja province where they

were collected from two different paramos.

Stereocaulon obesum Th. Fr., Stereoc. Piloph. Comm.: 28. 1857.

Spot test: K+ yellow, PD- or slowly yellow. Substances detected by TLC:

atranorin and lobaric acid.

SPECIMEN EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53’S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M. Prieto

(H-UTPL YG-0056).

Stereocaulon obesum is characterized its abundant and small upright and

thin pseudopodetia, unbranched at the base, but branched toward the apex,

bicolored peltate phyllocladia, and inconspicuous or irregular translucent

cephalodia (Rincén-Espitia & Mateus 2013).

Restricted to the Neotropics at 1700-4000 m altitudes in Costa Rica,

Ecuador, Guatemala, and Panama (Sipman 2002b), in Ecuador S. obesum was

previously found in Pichincha (Arvidson 1991). Our collection constitutes the

first record for Azuay province.

Stereocaulon ramulosum Raeusch., Nomencl. Bot., Ed. 3: 328. 1797.

Spot test: K+ yellow, PD+ slowly yellow. Substances detected by TLC:

atranorin and perlatolic acid.

SPECIMEN EXAMINED—ECUADOR, Loja. Saraguro: Loma del Oro, 3°40’52”S 79°14’24” W,

3245 m, paramo, 7.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0039).

Stereocaulon ramulosum is characterized by its >20 mm tall pseudopodetia

unbranched at the base with a few branches towards the apex and with few

more or less branched cylindrical phyllocladia, abundant large convex reddish

brown to black apothecia, and globose wrinkled cephalodia (Sipman 2002b,

Rincon-Espitia & Mateus 2013).

Widespread in the Southern Hemisphere and in Neotropical mountains

at 1000-4500 m altitudes in Brazil, Bolivia, Colombia, Costa Rica, Cuba,

Dominican Republic, Ecuador, Mexico, Peru, and Venezuela (Sipman

2002b), in Ecuador S. ramulosum was previously recorded from Chimborazo,

Loja, Pastaza, Pichincha, and Zamora-Chinchipe provinces (Mitten 1851,

Leighton 1866, Miller 1879, Zahlbruckner 1905, 1907, Noske & Sipman

2004). Our collection represents the first record for Loja province.

Stereocaulon tomentosum Fr., Sched. Crit. Lich. Suec. Exs. 3: 20. 1825.

Spot test: PD+ orange, K+ orange. Substance detected by TLC: atranorin

and stictic acid, corresponding to the type variety.

Terricolous lichens from Ecuador ... 171

SPECIMENS EXAMINED—ECUADOR, Azuay. Cuenca: Parque Nacional El Cajas,

2°46'53’S 79°13’21”W, 3930 m, paramo, 27.7.2010, A. Benitez, Y. Gonzalez & M. Prieto

(H-UTPL YG-0061); 2°47’03”S 79°13’30”W, 4009 m, paramo, 15.2.2014, A. Benitez & Y.

Gonzalez (H-UTPL YG-0308).

Stereocaulon tomentosum is characterized by an erect to prostrate thallus,

with lobed, squamulose, warty, or flattened phyllocladia (Brodo et al. 2001),

pseudopodetia covered with a thick felt layer (white tomentum) that also

covers the cephalodia, and numerous small (<1 mm diam) apothecia sitting

on short subapical branchlets and with spherical cephalodia (Sipman 2002b).

A subcosmopolitan species widespread in the Neotropics at 2300-4400

m altitudes in Bolivia, Colombia, Costa Rica, Dominican Republic, Ecuador,

Mexico, Peru, and Venezuela (Sipman 2002b), in Ecuador S. tomentosum was

previously found in Chimborazo, Pichincha, and Zamora-Chinchipe (Leighton

1866, Arvidsson 1991, Néske & Sipman 2004). Our collections are the first

records for Azuay province.

Thamnolia vermicularis (Sw.) Schaer., Enum. Crit. Lich. Eur.: 243. 1850.

Spot test: PD+ yellow-orange, K+yellow-orange, UV-, with thamnolic acid.

SPECIMENS EXAMINED—ECUADOR, Loja. Loja: Parque Nacional Podocarpus,

Cajanuma, 4°06’59”S 79°09’41” W, 3337 m, paramo, 29.6.2010, A. Benitez, Y. Gonzalez

& M. Prieto (H-UTPL YG-0077); Espindola: Jimbura, 4°42’37”S 79°25’46’W, 3450 m,

paramo, 8.7.2010, A. Benitez, Y. Gonzalez & M. Prieto (H-UTPL YG-0109); Azuay.

Cuenca: Parque Nacional El Cajas, 2°46’53”S 79°13’21”W, 3930 m, paramo, 27.7.2010,

A. Benitez, Y. Gonzalez & M. Prieto; 2°47’03”S 79°13’30”W, 4006 m, paramo, 15.2.2014,

A. Benitez & Y. Gonzalez (H-UTPL YG-0306); 2°47’36’S 79°12’09” W, 3890 m, 26.3.2014

(H-UTPL YG-0307).

Thamnolia vermicularis is characterizes as a fruticose species with a 20-70 x

1-2.5 mm decumbent to erect thallus with unbranched to slightly branched

hollow white to cream white stalks that are pointed at the tips, and which lacks

soredia, isidia, and fruiting bodies (Brodo et al. 2001).

The molecular study of Thamnolia Platt & Spatafora (2000) detected enough

genetic differences separating T’ vermicularis (containing thamnolic acid)

and T. subuliformis (containing squamatic and baeomycesic acids and with a

UV+Y cortex and a UV+ blue-white medulla) to support them as independent

species.

Widespread in Australia, Asia, Europe, New Zealand, and North America

(Smith et al. 2009), T: vermicularis has been recorded at 2800-4550 m

altitudes in the Neotropical region from Argentina, Bolivia, Chile, Colombia,

Costa Rica, Ecuador, Peru, and Venezuela (Soukup 1965, Sipman 1999,

Calvelo & Liberatore 2002, Flakus & Wilk 2006, Villagra et al. 2009). In

Ecuador, it was previously found in Chimborazo, Cotopaxi, and Pichincha

172 ... Gonzalez & al.

provinces (Zahlbruckner 1905, 1907; Arvidsson, 1991, Sklenay et al. 2010).

Our collections constitute first records of T. vermicularis for Azuay and Loja

provinces.

Acknowledgements

Financial support for this study was received from the Universidad Técnica

Particular de Loja (UTPL) and a scholarship to the first author provided by Secretaria

Nacional de Educacién Superior, Ciencia, Tecnologia e Innovacién (SENESCYT) of

Ecuador. We thank T. Ahti for the confirmation of some specimens and T. Ahti and

R. Pino-Bodas for their comments on the manuscript.

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MY COTAXON

January-March 2017— Volume 132, pp. 177-182

http://dx.doi.org/10.5248/132.177

Three lichen species in Buellia, Catillaria, and Cheiromycina,

new to Poland

MARTIN Kukwa!', PAWEE CZARNOTA? & ANNA LUBEK?

' Department of Plant Taxonomy and Nature Conservation, University of Gdansk,

Wita Stwosza 59, PL-80-308 Gdansk, Poland

? Department of Agroecology, University of Rzeszow,

Cwiklinskiej 1A, PL-35-601 Rzesz6w, Poland

° Institute of Biology, Jan Kochanowski University in Kielce,

Swietokrzyska 15A, PL-25-406 Kielce, Poland

CORRESPONDENCE TO *: dokmak@ug.edu.pl

ABsTRACT—Three sterile crustose lichen species, Buellia arborea from the Tatra Mts

and Catillaria fungoides and Cheiromycina reimeri from Bialowieza National Park, are

recorded as new to Poland. The record of Cheiromycina reimeri is also the first from Europe.

Characteristics of all three species, notes on similar taxa, distribution, and habitat preferences

are provided.

Key worps—sorediate lichens, lichenized fungi, Ascomycota

Introduction

Crustose, persistently or usually sterile lichens have been for a long time one

of the most neglected groups of species in the Polish lichen biota, especially

before thin layer chromatography (TLC) became a routinely used method in

their determination. However, the knowledge on their occurrences, distribution,

and ecology have been much improved in the past 15 years, including species

reported as either new to Poland or new to science (e.g., Kukwa & Kubiak 2007,

Czarnota et al. 2009, Kukwa & Jabtonska 2009, Czarnota & Guzow-Krzeminska

2010, Kukwa et al. 2012, Zdunczyk & Kukwa 2014, Guzow-Krzeminska et al.

2016).

In this paper, we present the first Polish records of three crustose lichen

species: Buellia arborea, Catillaria fungoides, and Cheiromycina reimeri.

178 ... Kukwa, Czarnota & Lubek

Material & methods

The material studied is deposited in following herbaria: GPN, FR, KTC, and UGDA.

The characteristics of taxa are based on Polish specimens, if not otherwise stated. Lichen

substances were studied by thin-layer chromatography (TLC) using the methods of

Orange et al. (2001). Reactions of the thalli with C (commercial bleach containing

sodium hypochlorite), K (10% solution of potassium hydroxide), and Pd (alcohol

solution of paraphenylenediamine) were also checked.

The localities examined are presented according to the modified ATPOL grid

square system (Kukwa et al. 2010, 2013 and literature cited therein). The numeration

of collecting plots in the Bialowieza National Park follows Falinski & Mulenko (1997).

The species

Buellia arborea Coppins & Tonsberg, Sommerfeltia 14: 111. 1992.

DESCRIPTION—THALLUS inconspicuous, white, endoxylic except for the

concave to convex, crateriform, punctiform to slightly elongated soralia

producing mostly dull, blackish-green pigmented soredia. SorED1IA single

or loosely aggregated, externally encrusted with crystals, which more or less

dissolve in K, composed of several algal cells surrounded by a stout, colorless

or greyish-green pigmented hyphal layer. APOTHECIA absent. Non-pigmented

soredia K+ yellowish, Pd-, C+ yellowish; pigment in soredial walls K+ dulling;

atranorin and placodiolic acid detected by TLC.

SPECIMEN EXAMINED—POLAND. Tatra Mts, High Tatra Mts, Tatra National Park,

forest unit no 53f, Dolina Rybiego Potoku valley, below Opalone Mt., 49°13’08.9”N

20°05’05.6”E, alt. 1330 m, ATPOL grid square Ge-60, Plagiothecio-Piceetum, on wood

of decaying log (Picea abies), 2.07.2010, P. Czarnota 6700 (GPN).

DISTRIBUTION & HABITAT—Although Buellia arborea is widespread in the

Holarctic, it is a rarely reported lichen, most probably due to its inconspicuous

thalli, which are usually sterile. It has been reported from Europe (Estonia,

Germany, Great Britain, Norway, Russia: Ural Mts, Slovakia, Sweden,

Switzerland, Ukraine), Asia (Russia: Siberia) and North America (Canada,

USA) (Dietrich et al. 1992, Tonsberg 1992, Foucard & Nordin 1999, Tonsberg

& McCune 2001, Bogdan 2002, Swensson et al. 2004, Lisicka 2005, Brackel &

Kocourkova 2006, Elvebakk & Bjerke 2006, Spribille & Bjork 2008, Coppins et

al. 2009, Suija et al. 2009, Urbanavichus & Andreev 2010, Davydov & Printzen

2012, Wirth et al. 2013).

In Poland B. arborea was found on the exposed wood of a spruce log in the

upper montane Carpathian spruce forest.

ComMENTS— The characteristic of Polish material of B. arborea agrees with the

species description in Tonsberg (1992). For a description of apothecia, which

were absent in the examined specimen, see Tonsberg (1992).

Three lichen species new to Poland ... 179

Buellia arborea resembles three other lichens—B. griseovirens (Turner &

Borrer ex Sm.) Almb., Violella fucata (Stirt.) T. Sprib., and Xylographa vitiligo

(Ach.) J.R. Laundon—all of which can also form crateriform soralia with

dark-colored soredia and may occupy the same habitat (decorticated logs).

All these species occur in Poland and differ in their secondary metabolite

compositions: B. arborea produces placodiolic acid and does not react with

Pd, whereas B. griseovirens contains norstictic acid (Pd+ yellow to orange);

V. fucata has fumarprotocetraric acid (Pd+ red); and X. vitiligo produces

stictic acid (Pd+ orange) (Tonsberg 1992, Wirth et al. 2013).

Lecanora orae-frigidae R. Sant. is another epixylic species with delimited

soralia producing dark outer soredia. It produces, however, usnic acid, zeorin,

and xanthones and occurs along the seashores in the arctic and boreal regions

of the Northern Hemisphere (Brodo & Vanska 1984).

Catillaria fungoides Etayo & van den Boom, Lichenologist 33: 107. 2001.

DESCRIPTION—THALLUS thin, not clearly delimited, whitish. SoRALIA

<0.25 mm in diam., dark brown to black, slightly convex, discrete or sometimes

confluent. SorEp1A globose, farinose, <20 um in diam., the outermost dark

brown to almost black, but inner soredia hyaline with colorless or pale brown

walls. APOTHECIA absent. Soredia C-, K- (blood-red crystals not formed

in squash-preparation) and Pd-; pigment in soredial walls also K negative;

material too scanty for TLC.

SPECIMEN EXAMINED—POLAND. NortH PopLasi£E District, Bielska Plain,

Bialowieza National Park, forest unit no 256, plot L06, 52°46’15”N 23°52’04”E, ATPOL

grid square Cg-55, Circaeo-Alnetum, on the bark of a hardwood log (Fraxinus excelsior),

19.08.2015, M. Kukwa 17314a, A. Lubek (KTC, UGDA).

DISTRIBUTION & HABITAT— Catillaria fungoides is an epiphytic lichen reported

from Africa (Cape Verde), Asia (South Korea, Turkey) and Europe (Czech

Republic, the Netherlands, Portugal, Slovakia, Spain) (Boom & Etayo 2001,

Boom et al. 2007, Malicek et al. 2014, Aptroot & Moon 2015).

In Poland C. fungoides was found on the bark of a hardwood log in a humid,

hardwood stand in the Bialowieza Forest. The species was sought by the first

authors in northern Poland several times; however, it could not be found there.

This suggests it is a rare lichen, perhaps confined to well-preserved forest

habitats.

ComMENtTS—The characteristics of Polish material of C. fungoides agree

with the species as described in Boom & Etayo (2001). For the description of

apothecia, which were absent in the examined specimen, see Boom & Etayo

(2001).

180 ... Kukwa, Czarnota & Lubek

Catillaria fungoides is characterized by dark brown to black soralia

(resembling sporodochia of some fungi) and the absence of lichen metabolites

detectable by standard TLC. This species can easily be confused with Rimularia

fuscosora Muhr & Tonsberg, which has larger (<0.6 mm diam.) soralia and

contains a trace of norstictic acid (Tonsberg 1992, Boom & Etayo 2001).

Norstictic acid can be difficult to detect in TLC when it occurs in trace

amounts, but blood-red crystals indicative of its presence are formed in squash-

preparations of soralia in K (Tonsberg 1992).

Dark soredia are also formed by Buellia griseovirens and Placynthiella

dasaea (Stirt.) Tonsberg, but they are never as dark brown or black as

in C. fungoides. Additionally, all three species differ in their chemistries:

C. fungoides lacks lichen metabolites detectable by TLC; B. griseovirens

produces atranorin and norstictic acid (sometimes only atranorin or only

norstictic acid); and P dasaea contains gyrophoric and lecanoric acids

(soredia C+ red) (Tonsberg 1992, Boom & Etayo 2001).

Cheiromycina reimeri Printzen, Nova Hedwigia 84: 262. 2007.

DESCRIPTION— THALLUs thin, crustose, contiguous, smooth, greyish green,

matt. CONIDIOMATA sporodochia, white, greyish-white or grey, discrete,

single or confluent, more or less ellipsoid, up to 0.80 x 0.50 mm, initially flat

to moderately convex. CONIDIOGENOUS CELLS terminal or lateral, globose or

broadly ellipsoidal, 7-10 x 7-8 um. Conrp1A palmate, 2-4 times dichotomously

branched, 20-30 um wide, 15-25 um tall, multi-septate, terminal branches

with 1-2 cells.

SPECIMENS EXAMINED—POLAND. NortTH PopLasiE District, Bielska Plain,

Bialowieza National Park, forest unit no 256, plot H10, 52°46’05”N 23°52’25”E, ATPOL

grid square Cg-55, Tilio-Carpinetum, on the bark of Carpinus betulus, 6.10.2015,

M. Kukwa 17681, A. Lubek (KTC, UGDA); plot 109, 52°46’08”N 23°52’20’E, Tilio-

Carpinetum, on bark of Carpinus betulus, 23.08.2015, M. Kukwa 17422, A. Lubek (FR,

KTC, UGDA).

DISTRIBUTION & HABITAT—Cheiromycina reimeri is a corticolous lichen that

has been known previously from Turkey and the Russian Far East (Sakhalin

Island) (Printzen 2007).

In Poland C. reimeri has been found only in Bialowieza National Park, twice

on the bark of Carpinus betulus. Our collections represent the first records of

the species in Europe.

ComMENTS—The characteristics of Polish material of C. reimeri agrees with

the species as described in Printzen (2007).

Cheiromycina reimeri can be confused with two other Cheiromycina species:

C. flabelliformis B. Sutton and C. petri D. Hawksw. & Poelt. Both, C. flabelliformis

Three lichen species new to Poland... 181

and C. reimeri have conidiogenous cells that are distinctly larger than the

conidial cells, but in C. flabelliformis, the conidiogenous cells do not exceed

9 um, whereas in C. reimeri they are consistently larger than 9 um. Additionally,

the terminal branches of the conidia in C. flabelliformis comprise up to 5 cells,

while in C. reimeri they are 1-3-septate (Printzen 2007). Cheiromycina petri

can be distinguished from C. reimeri by its conidiogenous cells, which are

not (or only slightly) larger than its conidial cells, whereas in C. reimeri, the

conidiogenous cells are always conspicuously larger (Printzen 2007).

Acknowledgments

We are very grateful to Dr Jurga Motiejunaité (Vilnius) and Dr Scott LaGreca

(Cornell University, USA) for reviewing the manuscript and for valuable comments

on a previous version of the paper. Dr Christian Printzen (Frankfurt) is thanked for

the confirmation of Cheiromycina reimeri. The research leading to these results was

funded by the Polish-Norwegian Research Programme operated by the National Centre

for Research and Development under the Norwegian Financial Mechanism 2009-2014

in the frame of Project Contract No. Pol-Nor/196829/87/2013 and the grant of Polish

Ministry of Science and Higher Education No. N N305 306835.

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MY COTAXON

January-March 2017— Volume 132, pp. 183-195

http://dx.doi.org/10.5248/132.183

Additions to the mycobiota of Poland

ANNA LUBEK! & MARTIN KUKWA?

' Institute of Biology, Jan Kochanowski University in Kielce,

Swietokrzyska 15A, PL-25-406 Kielce, Poland

? Department of Plant Taxonomy and Nature Conservation, University of Gdansk,

Wita Stwosza 59, PL-80-308 Gdansk, Poland

CORRESPONDENCE TO: ‘anna.lubek@ujk.edu.pl, *dokmak@ug.edu.pl

ABSTRACT—One saprobic fungus (Agyrium rufum), one facultative lichenicolous fungus

(Trimmatostroma quercicola), and six obligatory lichenicolous fungi (Arthonia coronata,

Cornutispora intermedia, Didymocyrtis melanelixiae, Minutoexcipula mariana, Stigmidium

rivulorum, and Weddellomyces xanthoparmeliae) are reported as new to Poland. Lecanora

pulicaris is a new host for Cornutispora intermedia and Ochrolechia turneri for Minutoexcipula

mariana. Seven of the reported species were found in natural forest communities in

Biatowieza National Park. Discussions on characteristics of each species and distributions

are also provided.

KEY worpDs—mitosporic fungi, Ascomycota

Introduction

Lichens, lichenicolous fungi, and allied fungi are a reasonably well-known

group of organisms in Poland (Faltynowicz 2003, Czyzewska & Kukwa

2009). However, additional research, especially in areas with well-preserved

natural forests, continue to provide new data on their occurrence, including

the discovery of species new to science (Kukwa & Diederich 2005, Kukwa &

Kubiak 2007, Kukwa & Flakus 2009, Zhurbenko et al 2009, Kukwa et al. 2010,

2012, 2013, Guzow-Krzeminska et al. 2016). Here we present the first Polish

records of eight species of saprobic and lichenicolous fungi.

Material & methods

The material studied is deposited in KTC, LBL, and UGDA. The technical

descriptions are based on Polish material, except for Didymocyrtis melanelixiae,

184 ... Lubek & Kukwa

where the teleomorph was not found among our samples. The localities are presented

according to the modified ATPOL grid square system (Cieslinski & Faltynowicz

1993, Kukwa et al. 2002). For species collected in Bialowieza National Park, the

numeration of collecting plots follows Falinski & Mutenko (1997).

The species

Agyrium rufum (Pers.) Fr., Syst. Mycol. 2: 232. 1822.

THALLUS endoxylic, non-lichenized, visible as paler patches of the

substratum with scattered, minute, flat to convex, irregularly roundish,

pale orange to red-brown apothecia. ExcIPLE soon excluded, but visible in

young apothecia, consisting of branched and anastomosing hyphae, pale to

deep orange-brown in section, K+ orange-red. HymMentum 70-80 um thick,

intensely I+ blue, K+ orange-red. ParaPpHysEs branched and anastomosing.

Asct broadly clavate, of Trapelia-type, 8-spored. AscosPporEs simple, hyaline,

10-12 x (4.5-)5-5.5 um.

Ecotocy—In Poland A. rufum was found only in Bialowieza National

Park (NE Poland) in two localities in natural pine-oak mixed forest growing

together with Micarea soralifera Guz.-Krzemin. et al. and other Micarea

species on decorticated logs. It is perhaps more common in Poland, but due to

the small size of apothecia easily overlooked in the field.

SPECIMENS EXAMINED—POLAND. NortTH PODLASIE DISTRICT, BIELSKA PLAIN,

Bialowieza National Park, forest unit no 256, plot G03, 52°46’02”N 23°51’48”E, ATPOL

grid square Cg-55, Pino-Quercetum, on decorticated log, 05.2015, M. Kukwa 15930,

A. Lubek (KTC, UGDA); plot G04, 52°46’02”N 23°51'53”E, Pino-Quercetum, on

decorticated log, 05.2015, M. Kukwa 15939, A. Lubek (KTC).

CoMMENTS— For a detailed description see also Dobson et al. (2009).

This is a non-lichenized, saprobic fungus that is superficially similar

to Micarea species with dark apothecia (e.g., M. denigrata (Fr.) Hedl. and

M. misella (Nyl.) Hedl.). Agyrium rufum is readily distinguished by its Trapelia-

type asci and the lack of photobiont in the thallus (Dobson et al. 2009).

Molecular phylogenetic analyses have shown that A. rufum is closely related

to Pertusaria DC. s. str., from which it is easily morphologically distinguished

by its much smaller apothecia and Trapelia-type asci (Hodkinson & Lendemer

2011).

Agyrium rufum has been found in many European countries growing on

wood of diverse woody plants in various forest ecosystems (e.g., Coppins &

Coppins 2006, Dobson et al. 2009, Mihal et al. 2012, Malicek & Palice 2013,

Spribille et al. 2014). The species has also been found in Australia (Kantvilas

2002).

Eight saprobic & lichenicolous fungi new to Poland ... 185

Arthonia coronata Etayo, Bull. Soc. Linn. Provence 47: 95. 1996.

ASCOMATA formed on discoloured parts of the host thalli, black,

convex, usually covered with simple, dark, <25 um long hairs giving the

surface a roughened appearance. HYMENIUM and HYPOTHECIUM hyaline.

ASCOSPORES 1-septate, 10-13 x 3.5-4 um, hyaline at first, becoming pale

brown with age.

EcoLtocy—In Poland A. coronata was recorded in Biatowieza National

Park where it is found on podetia of sorediate Cladonia species growing

on wood and bark of logs and stumps in pine-oak mixed forest and fresh

(spruce)-pine forest.

SPECIMENS EXAMINED—POLAND. NorTH PODLASIE DISTRICT, BIELSK PLAIN,

Bialowieza National Park, ATPOL grid square Cg—55, forest unit no 256, plot E09,

52°45'55"N 23°52’20"E, Pino-Quercetum, on Cladonia subulata, on stump, 08.2015,

A. Lubek (KTC); plot G01, 52°46’02”N 23°51'37”E, Pino-Quercetum, on C. coniocraea

growing on stump, 05.2015, M. Kukwa 15918, A. Lubek (KTC); plot G04, 52°46’02”N

23°51'53”E, Pino-Quercetum, on C. grayi growing on bark of log, 05.2015, M. Kukwa

15897, A. Lubek (KTC) and C. digitata growing on decorticated log, 05.2015, A.

Lubek (KTC); plot H04, 52°46’05”N 23°51’53”E Peucedano-Pinetum, on C. grayi

and C. coniocraea growing on decorticated log, 05.2014, M. Kukwa 13592, A. Lubek

(KTC, UGDA); plot L07, Querco-Piceetum, on C. grayi growing on decorticated log,

29.09.2015, M. Kukwa 17540a, A Lubek (UGDA); plot M01, 52°46’18”N 23°51'37’E,

Peucedano-Pinetum, on C. grayi growing on wood of log, 05.2015, A. Lubek (KTC).

ComMENTS—For descriptions see also Etayo (1996) and Coppins & Aptroot

(2009).

Arthonia coronata is distinguished by its black and convex ascomata

with usually simple dark hairs (best visible in apothecial section under the

microscope) giving the ascomata a roughened appearance, and the 1-septate

hyaline ascospores that become pale brown with age, ascospores. It cannot be

with confused with any other lichenicolous fungi inhabiting Cladonia thalli

(Etayo 1996).

The species was first found in France and Spain on soralia of Flavoparmelia

caperata (L.) Hale but later was also recorded from other countries on thalli of

Cladonia species. The presence of the fungus is manifested by a discolouration

of the host thallus by black ascomata (Etayo 1996, Coppins & Aptroot 2009,

Diederich et al. 2012).

Arthonia coronata has been found in Africa (Canary Islands), North

America, and Europe (Austria, Belarus, Belgium, France, Germany, Great

Britain, Italy, Luxemburg, Portugal, Spain, Sweden) (Etayo 1996, Coppins &

Aptroot 2009, Diederich et al. 2012, Boom 2013, Svensson et al. 2013, Roux et

al. 2016, Tsurykau et al. 2016).

186 ... Lubek & Kukwa

Cornutispora intermedia Punith. & D. Hawksw., Mycol. Res. 107: 920. 2003.

ANAMORPH—This species is known only in the anamorphic state, which

causes discolourations of host thalli. Pycnip1a scattered, immersed or with

upper parts exposed over the host thallus, globose with pseudoparenchymatous

walls, pale brown to dark brown, c. 150-160 um in diam. Conrp1a holoblastic,

hyaline, tri-radiate; main axes 9-12 um high (from the truncate bases to a

point where the conidia branch) and 2-2.5 um wide; divergent arms 3-5 x

2-2.5 um, with swollen bases. Both axes and arms end with narrow, 3-5 um

long, tubular structures or apical appendages.

EcoLtoGy—In Poland the species was found in one locality in Bialowieza

National Park on the thallus of Lecanora pulicaris (Pers.) Ach. in a pine-oak

mixed forest; L. pulicaris is a new host lichen for this fungus.

SPECIMEN EXAMINED—POLAND. NorTH PODLASIE DISTRICT, BIELSK PLAIN,

Bialowieza Forest, Biatowieza National Park, ATPOL grid square Cg—55, forest unit no

256, plot D03, Pino-Quercetum, on Lecanora pulicaris growing on bark of twigs of fallen

Picea abies, May 2015, A. Lubek s.n. (KTC).

CoMMENTS—For a full description of C. intermedia, see also Punithalingam

(2003).

Cornutispora intermedia is the third species of the genus reported from

Poland, the others being C. ciliata Kalb and C. lichenicola D. Hawksw. &

B. Sutton (Kukwa & Flakus 2009, Kukwa et al. 2013). In C. ciliata the main

conidial axes are shorter (usually 6.5-9 um long; in C. intermedia 8-11 um)

and the bases of the divergent arms are bulbous (only swollen in C. intermedia).

Cornutispora lichenicola has narrower main conidial axes (<2 um wide; in

C. intermedia >2 um) (Punithalingam 2003).

Cornutispora limaciformis Piroz. and C. pittii D. Hawksw. & Punith.

produce conidia of very similar in shape to those in C. intermedia, but

the main conidial axes are >13 um long in C. limaciformis and <3.5 um in

C. pittii. In addition, C. limaciformis is a non-lichenicolous fungus growing

on apothecia of Therrya fuckelii (Rehm) Kujala (Punithalingam 2003);

neither of these species has yet been found in Poland.

Cornutispora intermedia has also been reported from Europe (Germany,

Italy, Peru) and USA on thalli of Lecanora chlarotera Nyl., Ochrolechia sp.,

Phaeophyscia orbicularis (Neck.) Moberg, Physcia sp., Thamnolia vermicularis

(Sw.) Schaer., Xanthoria parietina (L.) Th. Fr., and X. polycarpa (Hoffm.)

Rieber (Punithalingam 2003, Brackel 2009, 2014, Etayo 2010, Schiefelbein et

al. 2014).

Eight saprobic & lichenicolous fungi new to Poland ... 187

Didymocyrtis melanelixiae (Brackel) Diederich, Harris & Etayo,

Fungal Div. 74: 74. 2015.

ANAMORPH causing discolourations of the host thalli. Pycnip1a

immersed in the host thallus, <90 um diam., macroscopically black; walls

pseudoparenchymatous, brown (darker around ostiola). Conrp1a_ broadly

ellipsoidal, rounded at both ends, smooth-walled, hyaline, with one large

(rarely two smaller) guttule, 4.5-5 x 3.5 um. SEXUAL STAGE not found.

EcoLtocy—lIn Poland the species has been found only at one locality in

southern Poland on thalli of Parmelia sulcata Taylor.

SPECIMENS EXAMINED—POLAND. RoztoczE SropKowE, Roztoczanski National

Park, Florianka settlement, 50°33’25”N 22°59’15”E, alt. c. 250 m, ATPOL grid square

Fg-01, on Parmelia sulcata growing on wooden fence, 17.09.2015, M. Kukwa 17532

(LBL, UGDA).

CoMMENTS—Only the anamorphic state was seen in the Polish material;

according to Ertz et al. (2015) ascomata are perithecial, the hamathecium

consists of paraphysoids, and asci are 8-spored. Ascospores are usually

2-septate, slightly constricted at the septa, brown, verruculose, 11.5-15 x 4-5.5

um. For full descriptions see also Brackel (2011) and Ertz et al. (2015).

Didymocyrtis melanelixiae in its anamorphic state can be confused with

four other species producing simple hyaline conidia and growing on lichens

of Parmeliaceae. The most common species, Briancoppinsia cytospora

(Vouaux) Diederich et al., differs in its larger conidia (5-7 x 1.5-2 um). Phoma

melanohaleicola D. Hawksw. & Earl.-Benn. develops pycnidia with seta-

like hyphae rising from the walls and does not damage the host thallus. In P

everniae D. Hawksw. the conidia measure 4.5-5.5 x 1-1.5 um, whereas in P.

dubia (Linds.) Sacc. & Trotter they are 3.5-5 x 1.5-2 um (Hawksworth & Cole

2004, Brackel 2011). Neither P. everniae nor P. dubia has been found in Poland.

Didymocyrtis melanelixiae is widespread, being recorded from Europe

(Austria, Belgium, France, Great Britain, Italy, Spain, Switzerland), North

America (Canada, USA), South America (Ecuador), and La Reunion, always

on members of Parmeliaceae Eschw. (Brackel 2011, Ertz et al. 2015, Hafellner

2015).

Minutoexcipula mariana V. Atienza, Biblioth. Lichenol. 82: 142. 2002.

CONIDIOMATA sporodochia-like, concave at first, but later becoming

confluent and convoluted. ExcrpLe distinct, thin, pseudoparenchymatic.

CONIDIOPHORES mostly unbranched, macronematous, conidiogenous cells

almost hyaline, 5-6 x 1.5-2 um with <3 annellations. Conrp14 6-7 um long,

brown, 1-septate, holoblastic, acrogenous, smooth-walled, with truncate bases.

188 ... Lubek & Kukwa

EcoLocy—Collected from the thallus of Ochrolechia turneri(Sm.) Hasselrot

growing on a fallen tree in a humid streamside within a black alder-ash forest

in Bialowieza National Park. It is the first record of Minutoexcipula mariana on

an Ochrolechia species. As Lepra, Ochrolechia, and Pertusaria are very closely

related (Schmitt & Lumbsch 2004), we consider this host range extension of

understandable.

SPECIMENS EXAMINED—POLAND. NorTH PODLASIE DISTRICT, BIELSK PLAIN,

Bialowieza National Park, forest unit no 256, plot O02, 52°46’24”N 23°51’42”E, ATPOL

grid square Cg-55, Circaeo-Alnetum, on Ochrolechia turneri growing on bark of fallen

Fraxinus excelsior, 10.2014, M. Kukwa 13086, A Lubek (KTC, UGDA).

CoMMENTS—For descriptions see also Atienza (2002) and Diederich (2003).

Minutoexcipula mariana can be confused with two other species growing

on members of Pertusariales: M. tuckerae V. Atienza & D. Hawksw. and

M. tuerkii Hafellner. Minutoexcipula tuckerae differs in its convex conidiomata

and 2-3-branched conidiophores, whereas M. tuerkii has larger conidia

and does not develop conidiophores (Atienza 2002, Diederich 2003).

Minutoexcipula tuerkii was reported from Poland by Kukwa & Flakus (2009).

Two Lichenodiplis species, L. pertusariicola (Nyl.) Diederich and

L. hawksworthii F. Berger & Diederich, are also similar, but both produce

closed conidiomata; additionally L. pertusariicola has aseptate conidia whereas

L. hawksworthii has larger conidia (8.5-13 x 5.5-5.5 um) (Diederich 2003). Of

these two, only L. pertusariicola has been recorded in Poland (Czyzewska &

Kukwa 2009).

Minutoexcipula mariana has previously been recorded in Asia (Japan, as

M. cf. mariana), Europe (Germany, Iceland, Italy, Portugal Spain, the Azores),

North America (Canada, USA, Mexico), and New Caledonia and only on

Lepra and Pertusaria species: L. albescens (Huds.) Hafellner, P. glomerata (Ach.)

Schaer., P. heterochroa (Mull. Arg.) Erichsen, P pluripuncta Nyl., P. xanthodes

Mull. Arg., and unidentified Pertusaria sp. (Atienza 2002, Diederich 2003,

Zhurbenko 2013, Brackel 2014, Zhurbenko et al. 2015).

Stigmidium rivulorum (Kernst.) Cl. Roux & Nav.-Ros.,

Bull. Soc. Linn. Provence 44: 449. 1994.

ASCOMATA numerous, <65 um diam., globose to subglobose. PARAPHYSES

absent; periphyses present around ostiola. Ascr 8-spored. ASCOSPORES

1-septate, hyaline, upper cell slightly wider and shorter than lower, 13.5-15 x

4—5 um.

Ecotocy—Collected in a small water spring in a shaded forest in northern

Poland. The species is perhaps more widespread, but due to the small

dimensions of perithecia it is easily overlooked.

Eight saprobic & lichenicolous fungi new to Poland ... 189

SPECIMEN EXAMINED—POLAND. GDANSK POMERANIA, WYSOCZYZNA

ZARNOWIECKA, Puzyckie Legi nature reserve, 54°38’N 17°51’E, ATPOL grid square

A3c-54, a small water spring in Circaeo-Alnetum, on Verrucaria hydrophila growing on

pebble, 12.08.2015, M. Kukwa 17069b (UGDA).

CoMMENTS—For full descriptions, consult Roux & Navarro-Rosinés (1994),

Molitor & Diederich (1997), and Zhurbenko & Hafellner (1999).

Stigmidium rivulorum is confined to freshwater Verrucaria and Staurothele

species. Three other Stigmidium species also grow on Verrucariaceae Eschw.:

S. clauzadei Cl. Roux & Nav.-Ros. has larger ascomata (80-130 x 65-120

um) and wider ascospores (5-6.5 um) and grows on non-aquatic Verrucaria

species (Roux & Navarro-Rosinés 1994); S. marinum (Deakin) Swinscow

is parasitic on marine Verrucaria growing on seashores and produces larger

ascomata (Swinscow 1965); and S. tetrasporum Etayo has 4-spored asci and

wider ascomata (80-150 um diam.), and was found on a sterile thallus probably

belonging to Verrucariaceae (Etayo 1994). Previously only S. clauzadei had

been found in Poland (Czyzewska & Kukwa 2009).

Stigmidium rivulorum has been found in Europe (Austria, Denmark: Faroe

Islands, Germany, Great Britain, Italy, Luxembourg) and Asia (Russia: Siberia,

South Korea) on thalli of Verrucaria aquatilis Mudd and other freshwater

species of Verrucaria Schrad. and Staurothele Norm. (Alstrup et al. 1994,

Molitor & Diederich 1997, Coppins & Coppins 1999, Zhurbenko & Hafellner

1999, Thiis & Dornes 2003, Orange et al. 2009, Brackel 2014, Berger & Turk

2015, Kondratyuk et al. 2015).

Trimmatostroma quercicola Diederich, U. Braun & Heuchert,

Bull. Soc. Nat. Luxemb. 111: 52. 2010.

HYPHOMYCETE facultative lichenicolous, growing on oak bark, often

overgrowing degenerate crustose lichen thalli or corticolous algae. COLONIES

an irregular net of blackish hyphae that are effuse, loose to dense, black, formed

by short, often agglomerated, irregular and branched conidial chains. CONIDIA

in simple or branched chains, globose to oblong, <4-septate (usually 3-septate),

reddish-brown in H,O, olivaceous-brown in K, <10 um wide, with <2 um thick,

rugose-rimulose to verrucose walls.

EcoLocy—In Poland Trimmatostroma quercicola was found on the bark of

old oak in moist oak-spruce forest in Bialowieza National Park. It is probably

widespread but overlooked due to the inconspicuous thalli.

SPECIMENS EXAMINED—POLAND. NorTH PODLASIE DISTRICT, BIELSK PLAIN,

Bialowieza National Park, forest unit no 256, plot E07, 52°45’55”N 23°52’10”E, ATPOL

grid square Cg—55, Querco-Piceetum, on degenerated lichen thalli growing on bark of

Quercus robur, 08.2014, M. Kukwa 14117 (KTC, UGDA).

190 ... Lubek & Kukwa

COMMENTS—For a complete description see also Diederich et al. (2010).

In general T: quercicola produces much thicker conidial chains

than the other Trimmatostroma species, which are easily visible under

stereomicroscope as an irregular blackish hyphal net. It also differs from

T: lecanoricola Diederich et al. by its orange-brown, K+ olivaceous conidia,

and from T: dendrographae Diederich et al. in producing brown conidia

(without a distinct olivaceous tinge). In T: hierrense Diederich & Ertz the

conidia often have darker outer walls on one side, whereas in T: glebarum

Brackel conidia are 0—-pluri-septate with cracking outermost wall layers,

and colonies formed of hyphae developing a visible dark net on or near the

surface of Flavoparmelia caperata thallus (Diederich et al. 2010, Brackel

2015). Trimmatostroma cetrariae Brackel has usually 1-septate conidia

(Zhurbenko & Brackel 2013), T. umbilicariicola Heuchert & U. Braun

develops multicellular and muriform segments in the fertile threads, and

conidia of T: varicellariae Heuchert & U. Braun are dark brown to black

(colour not changing in K) (Heuchert & Braun 2014).

The habit of T. quercicola (and also other lichenicolous Trimmatostroma

species) is similar to Intralichen spp., which differ by having smooth-walled,

paler, mostly 0-2-septate conidia and conidiophores usually that are completely

immersed in the host thalli or apothecia (Diederich et al. 2010). Taeniolella

species can be distinguished by conidia formed in acropetal chains and semi-

macronematous, superficial conidiophores; multicellular aggregations of

conidial cells are also absent in Taeniolella but are common in Trimmatostroma

species (Diederich et al. 2010).

This recently described species is known from Luxembourg and Belgium

(Diederich et al. 2012).

Weddellomyces xanthoparmeliae Calat. & Nav.-Ros., Mycotaxon 69: 505. 1998.

AscoMata black, perithecioid, scattered in the host thallus, which is

discoloured where infected. PERITHECIA arising singly or in small groups, at first

immersed, but later becoming partly exposed over the host thallus. Ascomatal

wall formed of cephalothecioid plates. Ascit 8-spored. HAMATHECIUM of

branched and anastomosing hyphae, AscosporEs are (1-)3-septate, brown,

finely granulose, 25-30 x 8-10 um.

EcoLocy—Found on the thallus of X. conspersa in the Sudety Mts, Poland.

SPECIMEN EXAMINED—POLAND. SuDETY Mrs, Gory SowlE MTs, near Jugow village,

50°38’51”N 16°30'19’E, elev. 720 m, ATPOL grid square Eb-95, on Xanthoparmelia

conspersa growing on rock, 08.2015, E. Ossowska s.n. (UGDA).

Eight saprobic & lichenicolous fungi new to Poland... 191

ComMMENTS—For other descriptions see also Calatayud & Navarro-Rosinés

(1998) and Kocourkova (1999).

Weddellomyces xanthoparmeliae is easily recognizable by its formation

of orange necrotic spots on the host thalli, an ascomatal wall formed of

cephalothecioid plates, usually 8-spored asci, and brown finely granulose

(1-)3-septate ascospores measuring 25-30 x 8-10 um. It differs from all other

Weddellomyces species in its smaller ascospores and host selection (Calatayud

& Navarro-Rosinés 1998). Other lichenicolous fungi with closed ascomata

growing on Xanthoparmelia species include Stigmidium neofusceliae Calat.

& Triebel, S. xanthoparmeliarum Hafellner, and Lichenostigma cosmopolites

Hafellner & Calat. (all distinguished by 1-septate ascospores), and Roselliniella

atlantica Matzer & Hafellner, which produces larger and usually aseptate

ascospores (Matzer & Hafellner 1990, Hafellner 1994, Calatayud & Triebel

1999, Hafellner & Calatayud 1999).

So far W. xanthoparmeliae is known from Europe (Czech Republic,

Germany, Spain) and North America. Hosts are usually Xanthoparmelia

species containing usnic acid: X. conspersa (Ehrh. ex Ach.) Hale, X. mexicana

(Gyeln.) Hale, X. cf. protomatrae (Gyeln.) Hale, and X. somloensis (Gyeln.)

Hale (Calatayud & Navarro-Rosinés 1998, Kocourkova 1999, Brackel 2007,

Kocourkova & Knudsen 2011), but Brackel (2014) reported it from thalli of

brown X. verruculifera (Nyl.) O. Blanco et al.

Acknowledgments

We are very grateful to Dr Jurga Motiejunaité (Vilnius) and Prof. Mark R.D.

Seaward (Bradford) for reviewing the manuscript and valuable comments on a

previous version of the paper. Dr Emilia Ossowska (Gdansk) is thanked for the

collection of Weddellomyces xanthoparmeliae and Dr Beata Krzewicka (Krakéw)

for the determination of Verrucaria hydrophila. The research leading to these

results has received funding from the Polish-Norwegian Research Programme

operated by the National Centre for Research and Development under the

Norwegian Financial Mechanism 2009-2014 in the frame of Project Contract No

Pol-Nor/196829/87/2013.

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MY COTAXON

January-March 2017— Volume 132, pp. 197-204

http://dx.doi.org/10.5248/132.197

Macrocyclic Edythea quitensis rust

on Berberis hallii in Ecuador

Maria E. ORDONEZ? & CHARLES W. BARNES?

' Pontificia Universidad Catélica del Ecuador, Escuela de Ciencias Bioldgicas,

Av. 12 de Octubre 1076 y Roca, Quito, Ecuador

? Instituto Nacional de Investigaciones Agropecuarias, Estacion Experimental Santa Catalina,

Panamericana Sur Km. 1 via Tambillo, Cantén Mejia, Ecuador

“ CORRESPONDENCE TO: meordonez@puce.edu.ec

ABSTRACT— The full macrocyclic autoecious life cycle of five spore stages was confirmed for

Edythea quitensis on Berberis hallii in the highlands of Ecuador. The spermogonial and aecial

stages had not previously been recorded. Spore descriptions and DNA sequence analysis are

included.

Key worps—Pucciniales, phylogeny, taxonomy

Introduction

Reports of rust occurring on Berberis in Ecuador date from 1891, when

Lagerheim collected and distributed specimens of two species that he labeled

as “Sphenospora quitensis’” and “Diorchidium berberidis’; subsequently,

Arthur (1918) published descriptions of these specimens as Uropyxis

quitensis and Sphenospora berberidis. Jackson (1931) proposed a new genus

Edythea to accommodate three South American species of rust on Berberis:

the two collected by Lagerheim (recombined as E. quitensis [the type species]

and E. berberidis (Lagerh. ex Arthur) H.S. Jacks.) and a new species from

Bolivia (E. tenella H.S. Jacks. & Holw.). Jackson (1931) depicts Edythea as

having urediniospores and teliospores “borne in a very characteristic and

presumably unusual manner” because of the lack of proper sori, as the

mycelial threads emerge from the stomata without rupturing the leaf tissue

to give rise to spores. The characteristics described by Jackson (1931) to

198 ... Ordofez & Barnes

differentiate between the three species of Edythea were the presence of

a vertical septum on teliospores for E. berberidis, a variable septum with

nearly globoid teliospores for E. quitensis, and a variable septum but with

ellipsoid teliospores for E. tenella. Urediniospores from the three species are

described as quite similar, but there is no mention of any spore stage, other

than urediniospores or teliospores, for the three Edythea species described

(Jackson 1931).

The host family of these rusts, the Berberidaceae, includes 13 genera,

of which only the genus Berberis L. is present in Ecuador. With over 500

species worldwide, Berberis (including Mahonia Nutt.) is the largest genus

of the family. There are two important centers of diversity for Berberis, one

in Eurasia with around 300 species mostly in China and the Himalayas and

another in South America with approximately 200 species (Ahrendt 1961).

The simple leaf type occurs throughout South America, while the compound

leaf type (previously referred to Mahonia) does not reach South America,

and is found only from Mexico to Costa Rica (Ulloa 2013). In Ecuador, 32

Berberis species have been reported, of which 16 are classified as endemic

(Ulloa 2013). All species occur in the highlands over 2000 meters above sea

level. The Berberis plant is used locally for coal, fencing, and firewood, with

the fruit used in jams and the roots used for medicinal purposes and staining

(Ulloa 2013).

In this paper we report the rust fungus Edythea quitensis found on

Berberis hallii Hieron. in Ecuador, as having a full macrocyclic autoecious

life cycle; sequence analysis of the internal transcribed spacer (ITS) region of

nuclear ribosomal DNA for three spore types also illustrates the phylogenetic

relationship between the Edythea rust samples collected on B. hallii in

Ecuador with rust collected from other Berberis species elsewhere.

Materials & methods

Collection and spore morphology

Rust infected leaves were collected from Berberis hallii in Quito, Pichincha

province. Fresh spores were examined microscopically in water, and teliospores,

urediniospores, and aeciospores were measured using Infinity Analysis software

(Lumenera Corp., ON, Canada). Twenty random spores from each collection

were measured for each spore type. Germinating teliospores, basidiospores,

and spermogonia were also observed. Collections were deposited in the QCAM

fungarium at Pontifica Universidad Catdlica del Ecuador, Quito, Ecuador (TABLE 1).

Molecular analysis

DNA was extracted separately from aecia/spermogonia and uredinia/telia found

on different collections of B. hallii leaves. The tissue was macerated with zirconium

Edythea quitensis on Berberis hallii (Ecuador) ... 199

TABLE 1. Edythea quitensis samples analyzed

SORT EEN GEOGRAPHICAL ALTITUDE SPORE TYPE GENBANK

COORDINATES [m a.s.l.] OBSERVED* NO.

QCAM6449 -0.18646, -78.46178 2890 II, Il KX298148

QCAM6450 -0.17609, -78.46702 2917 0, I, U, UI, IV KX298149

QCAM6451 -0.17455, -78.46968 2879 II, Il KX298150

QCAM6452 -0.17711, -78.46658 2919 II, Il KX298151

QCAM6453 -0.17497, -78.46890 2890 II, Il KX298152

QCAM6454 -0.17708, -78.46670 2887 0, I, II, III, IV KX298153

* Following Cummins & Hiratsuka 1983

TABLE 2. Rust sequences from GenBank included in the phylogenetic analysis

GENBANK NO. TAXON COUNTRY Host REFERENCE

JX047488 Puccinia sp. Sweden Berberis sp. Berlin et al. 2013

JX047486 Puccinia sp. Sweden Berberis sp. Berlin et al. 2013

JX047489 Puccinia sp. Sweden Berberis sp. Berlin et al. 2013

JX047484 Puccinia sp. Sweden Berberis sp. Berlin et al. 2013

JX047456 P. graminis f.sp. tritici Sweden Berberis sp. Berlin et al. 2013

JX047465 P. graminis f.sp. tritici Sweden Berberis sp. Berlin et al. 2013

JX047468 P. graminis f.sp. tritici Sweden Berberis sp. Berlin et al. 2013

JX047467 P. graminis f.sp. tritici Sweden Berberis sp. Berlin et al. 2013

JX047477 P. graminis f.sp. avenae Sweden Berberis sp. Berlin et al. 2013

JX047478 P. graminis f.sp. avenae Sweden Berberis sp. Berlin et al. 2013

JX047482 P. graminis f.sp. avenae Sweden Berberis sp. Berlin et al. 2013

JX047475 P. graminis f.sp. avenae Sweden Berberis sp. Berlin et al. 2013

AY956558* P. striiformoides Iran Dactylis glomerata Abbasi et al. 2004

DQ417406* P. striiformis f.sp. tritici USA Triticum aestivum Barnes & Szabo

2007

GU382672 P. striiformis f.sp. tritici USA Berberis chinensis Jin et al. 2010

GQ457304+ P. pseudostriiformis USA Berberis sp. Jin et al. 2010

GQ457306+ P, pseudostriiformis USA Berberis koreana Jin et al. 2010

* Best Blastn hits (from non-Berberis hosts) to Edythea quitensis ITS sequence

+ Formerly Puccinia striiformis f.sp. poae (Jin et al. 2010)

beads and soaked in 0.5% SDS lysis buffer at 55°C for 3 hours. Protein precipitation

was done with 4M guanidine isothyocianate solution, pH 7.5, followed by isopropanol

and ethanol precipitation. The ITS (ITS1+5.8S+ITS2) region was amplified with

primers ITSIF (Gardes & Bruns 1993) and ITS4BR (Vialle et al. 2009). The PCR

cocktail comprised 0.2 uM of each primer, 1X PCR buffer, 3mM MgCl, 0.2mM

dNTPs, 0.1 U/uL Platinum Tag DNA polymerase (Invitrogen, Brazil), and Ing/uL of

DNA. Amplification conditions were: 94°C for 3 min., 30 cycles of 94°C for 30 s, 50°C

200 ... Ordofez & Barnes

for 30s, and 72°C for 60 s, anda final extension at 72°C for 10 min. PCR products were

sequenced at Macrogen Inc. (Seoul, South Korea). Sequences were edited in Geneious

R9 (Kearse et al. 2012) and submitted to GenBank (http://www.ncbi.nlm.nih.gov).

The alignments were deposited into TreeBASE (http://purl.org/phylo/treebase/

phylows/study/TB2:S19447).

Sequence data were analyzed using Maximum Likelihood with a Custom

(010212) Substitution model determined by jModelTest 2.0.2 (Darriba et al. 2012,

Guindon & Gascuel 2003) according to Corrected Akaike Information Criterion

(AICc). Sequence data available on GenBank from rust fungi infecting Berberis

species from other countries were included in the analysis, as well as the two best

Blastn hits (TABLE 2). A bootstrap of 1000 replicas was used.

Taxonomy

Edythea quitensis (Lagerh. ex Arthur) H.S. Jacks. & Holw.,

Mycologia 23: 99. 1931. Fic. 1

= Uropyxis quitensis Lagerh. ex Arthur, Bot. Gaz. 65: 464. 1918.

Type: Ecuador, Quito, stages II and III on Berberis sp., April 1891, G. Lagerheim

(PUR 68850, holotype, originally deposited as “Sphenospora quitensis”).

Spermogonia, aeciospores, urediniospores, teliospores, and basidiospores

were observed on B. hallii in Quito. A total of six rust samples were analyzed,

two showing all five spore stages and four only urediniospores and teliospores

(TaBLE 1). Spermogonia and aecia were found occurring simultaneously on

leaves several times during the year, usually after rains.

Spermogonia were observed on the upper side of the barberry leaves, and

aecial cups were present on the underside of the leaves. Spermogonia were

yellow initially, becoming more orange in color as they aged. Aecial cups were

bright orange. Aeciospores were ellipsoid to globose, 27.3-21.5 x 27.0-20.9

um, verrucose, with a thin wall, yellow-orange in color. Urediniospores and

teliospores were found year round and matched characteristics described

by Jackson (1931); specifically: both spore types emerged through the

stomates without rupturing the tissue and were cinnamon-brown in color;

urediniospores were mostly globoid, single-celled, 29.5-22.8 x 28.2-22.6

um; teliospores were globoid, two-celled, 27.6-22.0 x 31.6-24.0 um with

pedicels 21.4-30.0 x 5.5-6.6 um. Germinating teliospores were observed

forming a basidium with four basidiospores on sterigmata.

Phylogeny

ITS sequence analyses obtained separately for aecia/spermogonia and for

uredinia/telia supported all within the same species (Fic. 2). The Ecuadorian

samples were distinct from several rust species reported from Berberis

Edythea quitensis on Berberis hallii (Ecuador) ... 201

Fic. 1. Edythea quitensis: macrocyclic stages. A. basidia; B. basidiospores on sterigmata;

C. spermogonia; D. spermogonial infection on upperside of barberry leaf; E. aeciospores;

FE. aecial cup; G. closeup of aecial cups on underside of barberry leaf; H. aecial infection on

underside of barberry leaf; I. teliospore (dashed arrow) and urediniospore (whole arrow);

J. mix of urediniospores and teliospores; K. urediniospore and teliospore infections on

underside of barberry leaf.

from Sweden and the USA. The resulting topology was well supported by

bootstrap values.

Discussion

The complete macrocyclic autoecious life cycle for Edythea quitensis

occurring on B. hallii is described here for the first time. The three original

202 ... Ordofiez & Barnes

E. quitensis KX298148 uredinia/telia

E. quitensis KX298149 aecia/spermogonia

E. quitensis §X298150 uredinia/telia

E. quitensis KX298151 uredinia/telia

E. quitensis KX298152 uredinia/telia

E. quitensis KX298153 aecia/spermogonia

P. striiformoides AY 956558

109 | Pst DQ417406

Pst GU382672

P. pseudostriiformis GQ457306

P. pseudostriiformis GQ457304

Puccinia sp. JX047484

Puccinia sp. JX047486

Puccinia sp. JX047488

Puccinia sp. JX047489

Pga JX047477

Pga JX047478

Pga JX047475

Pga JX047482

Pgt JX047456

Pgt JX047465

Pgt JX047468

Pgt JX047467

0.04

Fic. 2. Maximum Likelihood phylogenetic tree based on the ribosomal DNA sequence of the

ITS1+5.8S+ITS2 region of Edythea quitensis and representative taxa of rust from Berberis spp., plus

the two best Blastn hits not found on Berberis spp. One thousand bootstrap replicates were used;

values 270% are given above branches.

Taxonomic abbreviations: Pst = Puccinia striiformis f.sp. tritici; Pga = P. graminis f.sp. avenae;

Pgt = P. graminis f. sp. tritici.

Edythea rusts on Berberis and a fourth species, E. soratensis Ritschel

(described from a 1920 collection on Berberis in Bolivia; Berndt et al. 2007),

have not previously been subjected to repeated seasonal field observations.

However, the increased attention being paid to Berberis spp. as potential

Edythea quitensis on Berberis hallii (Ecuador) ... 203

alternate hosts for stripe rust Puccinia striiformis Westend. in wheat growing

areas (Jin et al. 2010, Zhao et al. 2013) afforded the opportunity for closer

examination of the rust species occurring on B. hallii in Quito. It is likely that

the original description of E. quitensis did not include the spermogonial and

aecial stages due to the restricted time and number of collections that were

available at that time.

The use of ITS sequence comparison of the different spore stages confirmed

that the aeciospores, urediniospores, and teliospores analyzed belong to the

same species.

Ecuador is within the center of diversity of Berberis species in South

America, and more work is needed to describe the rust fungi that infect

these hosts. Rust species description and identification are important for

managing wheat rust resistance in Ecuador and adjacent regions as well as

for documenting fungal biodiversity in these countries.

Acknowledgments

Funding for this work was provided by Pontificia Universidad Catolica del Ecuador.

We wish to thank Dave McLaughlin and Cathie Aime for reviewing this manuscript,

and Shaun Pennycook for his suggestions and assistance of this work.

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MY COTAXON

January-March 2017— Volume 132, pp. 205-216

http://dx.doi.org/10.5248/132.205

Leucoagaricus ariminensis sp. nov., a lilac species

from Italy

E Dovana’, M. Contv?, P- ANGEL’, A. BRANDI? & M. MUCCIARELLI!

‘Department of Life Sciences and Systems Biology, University of Torino,

Viale P.A. Mattioli 25, 10125 Torino, Italy

? Via Marmilla, 12 (I Gioielli 2), 07026 - Olbia (OT), Italy

> Via Cupa 7, 47923 - Corpolo di Rimini (RN), Italy

* Viale Paolo Mantegazza 41, 47921 - Rimini (RN), Italy

“ CORRESPONDENCE TO: francescodovana@libero.it

ABSTRACT—A new species, Leucoagaricus ariminensis, collected under Cupressus

sempervirens in a park in Rimini (central Italy), is described and illustrated. The new species

is compared with the other known lilac-coloured lepiotoid species, and its phylogenetic

position, based on nrITS sequences, is determined. Colour photographs and illustrations of

diagnostic anatomical characters are also provided.

Key worps—Agaricaceae, Agaricales, lepiotaceous fungi, rDNA, taxonomy

Introduction

The genus Leucoagaricus Locq. ex Singer is considered to have a widespread

geographical distribution even if it might appear more commonly linked to the

tropical and sub-tropical zones of Africa and America (Singer 1986). Data in

Vellinga (2004) indicate that the number of Leucoagaricus species in Europe

increases as one moves southwards from the northern areas. Citations from the

checklist of Italian fungi (Onofri et al. 2005) and other sources (Consiglio &

Contu 2004, Zotti et al. 2008) establish occurrences of at least 52 Leucoagaricus

species in Italy, a figure significantly higher than the numbers recorded in

northern European areas (Vellinga 2004, Lange 2008). Of all species described

in Italy, 58% occur in the north, 62% in the south (with islands included),

and the majority, 73%, in the central regions. Considering only the non-

206 ... Dovana & al.

coastal northern regions, the number of species decreases to less than 40%,

demonstrating the greater biodiversity of Leucoagaricus in milder climatic

zones.

Numerous new species have been described in recent years (Ge 2010; Liang

et al. 2010; Vellinga & Balsley 2010; Vellinga et al. 2010; Munoz et al. 2012,

2014; Kumari & Atri 2013; Ye et al. 2014; Ge et al. 2015), a clear indication that

the exact number of species in this genus is still incomplete. Some have been

found in urban parks showing that these areas can be an interesting habitat for

the study of this genus.

During a recent fungal biodiversity study conducted in Parco XXV Aprile,

a public park in Rimini, a Leucoagaricus species was found, characterized

by a dark violaceous pileus, violet/lilac tinged stipe and annulus rim, rather

fleshy basidiomata, negative reaction to ammonia vapours, and white flesh that

yellowed when handled. Comparison with the known Leucoagaricus species

and ITS sequence analyses indicated that this species represents a novel taxon,

which we describe below.

Materials & methods

Morphology

The macroscopic descriptions are based on observations of fresh material, which

was photographed with a Canon EOS 60D camera fitted with a Canon EF-S 60 mm

macro lens. The specimens were dried with an electric dryer. Voucher specimens

were deposited in the Herbarium, Museo di Storia Naturale di Venezia, Venice, Italy

(MCVE). The terminology used in the descriptions follows Vellinga (2001). Basidioma

colours were coded according to Munsell (1994). The micro-morphological characters

are based on the study of dried material rehydrated in KOH or NH,OH; other reagents

used were Congo Red for staining cell-walls, anionic phloxine for colouring the

cytoplasm, Cresyl Blue for testing the metachromatic reaction of the spores, Melzer’s

reagent to detect spore dextrinoid reactions, glycerine tampon L4 for the observation

of pigments, Chlorazol Black (L4C) for contrasting the septa. Spore nuclei were

stained with DAPI (1 mg/mL) and observed with a fluorescence microscope. All

observations and measurements were carried out with an optical trinocular Optrech

B5 microscope, with x10, x25, x40, x60 lenses and a x100 immersion lens. Micro-

photos were taken with a Canon EOS 60D camera connected to the microscope with an

extension tube. At least 20 basidia, basidiospores, cystidia and terminal elements of the

pileipellis were measured. Basidiospores were measured both from fresh spore deposit

and from dried specimens. Basidiospore dimensions are expressed as (a) b-c (d),

where (a) = minimum value, b = (average - standard deviation), c = (average +

standard deviation), and (d) = maximum value. The following abbreviations are used:

L = number of lamellae, and 1 = number of lamellulae; [x/y/z] indicates that altogether

x spores, from y samples, from z collections were measured.

Leucoagaricus ariminensis sp. nov. (Italy) ... 207

DNA extraction, PCR amplification, DNA sequencing

The DNA was extracted with NaOH (Osmundson et al. 2013); 2 mg of dry sample

were homogenized in 250 ul of 0.5M NaOH with a pestle. After 10 minutes to allow

for sedimentation, 5 ul of the extract was removed and diluted in 195 ul of 100mM

Tris-HCl at pH 8.0, and 1 ul of the dilution was used as template DNA. The ITS region

was amplified with primers ITS1F (Gardes & Bruns 1993) and ITS4 (White et al. 1990).

PCR was performed in a 25 ul reaction volume following Gardes & Bruns (1993).

PCR products were purified and sequenced by IGA Technology Services (Udine,

Italy).

Sequence alignment & phylogenetic analysis

Sequences were assembled and edited in Geneious v. 8.1.2 (Kearse et al. 2012) and

then submitted to GenBank (see Fic. 1 for accession numbers).

The initial BLASTn results revealed that all reference sequences with identities

>85% (and E = 0) to L. ariminensis belong to the Piloselli clade, following criteria

reported in Vellinga (2010), Vellinga et al. (2010, 2011), and Mufioz et al. (2012, 2014).

Our dataset includes sequences in /Piloselli from previous phylogenetic studies

(Vellinga 2010; Vellinga et al. 2010, 2011; Munoz et al. 2012, 2014) and other

sequences with greatest similarity available in GenBank. We also included a sequence

of L. ionidicolor (sequenced in this work, KU953373) collected in the Czech Republic

and described in Holec (2009) and added for comparison six sequences representing

Leucoagaricus sect. Rubrotincti. Cystolepiota seminuda was used as outgroup.

The sequences were aligned using MAFFT v 7.017 (Katoh et al. 2002) with default

conditions for gap openings and gap extension penalties.

Phylogenies were inferred with RAXML v.7.2.8. (Stamatakis 2006) and MrBayes v.

3.2.4 (Huelsenbeck & Ronquist 2001) using jModelTest v. 2.1.7 (Darriba et al. 2012)

to choose best-fit models of nucleotide substitution. The GTR+G model was chosen

for both analyses.

A total of 1000 bootstrap replicates were performed to assess the relative robustness

of the branches of maximum likelihood in RAxML. In Bayesian analysis, MCMC was

performed for four chains and run for 1,0000,000 generations (sampling trees every

1000 generations) with a 25% burn-in.

Taxonomy

The alignment includes 96 ITS sequences representing 28 species of white-

spored Agaricaceae and comprises 805 characters.

Both Maximum Likelihood and Bayesian analyses produced the same

topology. Only the Bayesian tree with posterior probability and bootstrap values

is shown (Fic, 1). Leucoagaricus ariminensis is a sister species of Leucoagaricus

ionidicolor (MLB = 100; BPP = 1), with which it shares 91.7% of bases/residues.

They form the sister clade of Lepiota decorata and are placed within /Piloselli.

The sequence of L. ionidicolor from the Czech Republic specimen shares

658/663 (99%) nucleotides compared with the Dutch sequence (AY 176415).

208 ... Dovana & al.

1/66

0.99/68

1/99

0.98/50

0.997

oi Lepiota fuliginescens AY243642

Leucoagaricus sp AY176440

Lepiota flammeotincta GU136171

Lepiota flammeotincta GU136170

Lepiota flammeotincta GU136169

Lepiota flammeotincta GU136167

Lepiota flammeotincta GU136166

Lepiota flammeotincta GU13616$

Lepiota flammeotincta GU136164

Lepiota flammeotincta GU136163

Leucoagaricus erythrophaeus GQ2S8472

Leucoagaricus erythrophaeus GQ258469

Leucoagaricus erythrophaeus GU136179

Leucoagaricus erythrophaeus GU136177

Leucoagaricus erythrophaeus GQ258471

Leucoagaricus erythrophaeus GQ258468

Leucoagaricus erythrophaeus GQ203803

Leucoagaricus erythrophaeus AY243644

Leucoagaricus decipiens GQ203803

00F Leucoagaricus pyrrhophaeus GU136199

ut Leucoagaricus pyrrhophaeus GQ258473

/78 raricus sp GU136200

VP Leucoagaricus flammeotinctoides GU136175

1/94) ub) * Leucoagaricus flammeotinctoides GU136174

Leucoagaricus flammeotinctoides AY243620

OOF Leucoagaricus pyrrhulus GU136201

Leucoagaricus pyrrhulus GQ2S8474

2100 Leucoagaricus pardalotus GU136202

Leucoagaricus pardalotus GQ258479

00) Leucoagaricus sp AY243633

Leucoagaricus brunnescens GQ203804

79 100° Leucoagaricus georginae GU136198

Leucoagaricus georginae AY243634

Leucoagaricus jubilaei AY 243635

Leucoagaricus variicolor 3X880033

N00F Leucoagaricus variicolor X880032

Leucoagaricus variicolor 3X880031

Leucoagaricus varlicolor JJX880030

Lepiota fuliginescens GU136189

Lepiota fuliginescens GU136186

0.9916

197

Lepiota fuliginescens AY2436A\

Lepiota fuliginescens GU136184

Lepiota fuliginescens GU136183

Lepiota fuliginescens GU136188

Lepiota fuliginescens AY243639

Piloselli

Lepiota fuliginescens GU136187

Lepiota fuliginescens AY243638

Lepiota fuliginescens AY243636

Leucoagaricus badhamii AY243643

woo) Leucoagaricus ariminensis KU953372

Leucoagaricus ariminensis KU953371

Leucoagaricus ariminensis KU953370 TYPE

0% Leucoagaricus ionidicolor KU953373

Leucoagaricus ionidicolor AY 176415

100° Lepiota decorata GU136197

Lepiota decorata AY243645

wis Lepiota roseilivida AY 176394

“| — Leptota roseilivida FFI21815

64 Lepiota roseilivida AY243646

100) U Leucoagaricus marriageae GQ329049

0.989 Lepiota roseilivida AF482866

Lepiota roseilivida EF121816

00° Leucoagaricus croceobasis KP853413

Leucoagaricus croceobasis K¥8$3412

1/100

Leucoagaricus sp GU136182

Leucoagaricus badhamii GQ329056

OOF Leucoagaricus dyscritus GU136181

Leucoagaricus dyscritus GU136180

Leucoagaricus adelphicus GU136190

Leucoagaricus adelphicus GQ258478

197|_ Leucoagaricus adelphicus AY243625

Leucoagaricus adelphicus AY243624

Leucoagaricus adelphicus AY 243623

Leucoagaricus adelphicus AY243622

Leucoagaricus adelphicus AY24362\

Leucoagaricus pilatianus GQ329057

Leucoagaricus pilatianus GQ329040

Leucoagaricus pilatianus A¥243626

/L00

0OF Leucoagaricus hesperius GU139790

Leucoagaricus hesperius GU139789

Leucoagaricus ef cupresseus AY243632

Leucoagaricus cf cupresseus AY243627

Leucoagaricus cupresseus GU136194

163 Leucoagaricus cupresseus AY 243628

Leucoagaricus cupresseus GU136195

Leucoagaricus cupresseus GU136191

19 Leucoagaricus cupresseus AY243631

Leucoagaricus cupresseus AY243630

Leucoagaricus cupresseus AY243629

1/100)

2F~ Leucoagaricus rubrotinctus JN944081

1/100 Leucoagaricus rubrotinctus F3481050

Leucoagaricus rubrotinctus N944082

Leucoagaricus sublittoralis AY 176442

Leucoagaricus wichanskyi AF482874

Leucoagaricus littoralis GQ329041

Rubrotincti

1/100

Cystolepiota seminuda AY176350

Fic. 1. Bayesian phylogram obtained from the general ITS sequence alignment of Leucoagaricus/

Lepiota spp.; Cystolepiota seminuda was used as outgroup taxon. Only BPP values >0.95 (in bold)

and MLB values >50% are shown above clade branches. Sequences of Leucoagaricus ariminensis

are in bold.

Leucoagaricus ariminensis sp. nov. (Italy) ... 209

Fic. 2. Leucoagaricus ariminensis (holotype, MCVE 28443). Basidiomata. Scale bar = 20 mm.

Leucoagaricus ariminensis Dovana, Angeli, Contu & Brandi, sp. nov. FIGS 2-4

INDEX FUNGORUM IF552049

Differs from Leucoagaricus ionidicolor by its more robust basidiomata, stipe that is

usually shorter than the pileus diameter, more variably shaped and larger cheilocystidia,

larger spores, and longer basidia.

TYPE: Italy, Emilia-Romagna, Rimini(RN), Parco XXV Aprile, along an avenue of cypress

trees, 27.12.2014, legit L. Brandi (Holotype, MCVE 28443; GenBank KU953370).

EryMotoay: the specific epithet “ariminensis” derives from Ariminum, Latin name for

Rimini, a city in Emilia Romagna Region of the Italian Republic.

Pileus 3.5-8 cm, initially convex, becoming plano-convex, then depressed

with age and plane in the centre; margin regularly involute at first, becoming

incurved for a long time, then finally extended and somewhat irregularly

sinuate; violet grey, violet brown-grey, violet (5P 3/4-8; 5P 4/4-8), tomentose,

with tufts of thick hairs forming fine, densely packed squamules (lens), sparser

at the margin and there the white flesh showing through. Negative reaction to

aqueous ammonia.

Stipe 2-4(-4.5) x 1-2 cm, cylindrical, shorter than the pileus diameter,

tapering upwards from an enlarged but not bulbous base, tapering downwards

in some specimens, base well-covered with leaf-mould or debris, fibrous,

fistulous; surface covered with white hairs faintly tinged with violet (5P 7/2-4)

210 ... Dovana & al.

. ihe eye

Fes t GN

ai eNO IVE

LGP4 SORRY

PAIN

Fic. 3. Leucoagaricus ariminensis (holotype, MCVE 28443). A, B. Cheilocystidia. C. Spores.

D. Terminal elements of the pileipellis. E. F Basidia. Scale bars = 10 um.

above the annulus, whitish and stained purple below (5RP 5/2-4)) where

brown stains appear with age or handling. Annulus pendant, membranous,

tomentose on the underside like the stipe surface, rim concolorous with the

pileus.

Lamellae, L = 70-90, | = 2-3, moderately crowded, rounded and free at

the stipe with a collarium, interspersed with numerous truncate lamellulae of

various lengths; white to pale cream; edge sterile.

Flesh thick, tender, fragile, fibrous in the stipe, yellowing when handled

or with age; odour faint, acidulous, only perceptible in young specimens,

becoming unpleasant and smelling of fish with age; taste mild. Non-staining

in contact with ammonia. Spore deposit light cream (2.5YR 8/1).

Basidiospores [270/3/4] (5.5-)6.5-8.1(-10.5) x (3.3-)4.1-4.9(-6.1) um,

on average 6.9-7.8 x 4.3-4.6 um, Q = (1.18-)1.45-1.84(-2.31) Q. = 1.5-1.8;

Leucoagaricus ariminensis sp. nov. (Italy) ... 211

10 um

Fic. 4. Leucoagaricus ariminensis.

a Spores. b Basidia. c Cheilocystidia. d. Terminal elements of the pileipellis. e. Caulocystidia.

Scale bars: a = 5 um; b-e = 10 um. (Drawings by Maria Tullii).

212 ... Dovana & al.

mainly ellipsoid, with a rounded or sometimes subacute apex, some

amygdaliform or subcylindrical, smooth, guttulate, thick-walled, without

a germ pore or suprahilar depression, dextrinoid, metachromatic in Cresyl

Blue, binucleate (rarely tetranucleate). Basidia 33.9-38.6 x 7.0-8.8 um,

4-spored, some 2-spored, seldom 1-spored, at times somewhat swollen in the

upper part. Lamellar trama + regular, hyphae 5-10 um wide, without clamps.

Pleurocystidia not observed. Cheilocystidia 48-73 x 12-15 um, abundant,

variable in shape, from cylindrical, clavate with an obtuse apex to lageniform,

rarely bifurcate. Pileipellis formed by repent interwoven hyphae, becoming a

trichoderm near the squamules, with long, apically rounded terminal elements

intermingled with elements that are tapered towards the apex, 150-330 x

7-12 um; vacuolar pigment brownish-green-grey, at the centre some hyphae

with granular vacuolar pigment; greenish-yellow excretory hyphae present.

Non-gelatinised hyphae at disc. Subpellis formed by + parallel hyphae,

9-16 (20) um wide, slightly interwoven, with apically rounded, clavate and

cylindrical terminal elements. Caulipellis formed by hyphae 5.9-13.8 um

wide, + parallel, the outer ones interwoven with apically rounded, clavate and

cylindrical terminal elements, some hyphae ending with long hairs; branched

hyphae and some with excrescences also observed; caulocystidia lageniform,

cylindrical, clavate, trunco-conical, 74-130 x 11-40 um. Clamps absent.

ECOLoGy: gregarious on bare sandy soil rich in vegetable litter (and animal

excrements) under old cypress trees, Cupressus sempervirens L. In winter,

from a single locality in Emilia Romagna.

ADDITIONAL MATERIAL STUDIED: ITALY, EMILIA-ROMAGNA, Rimini (RN),

Parco XXV Aprile: along an avenue of cypress trees, 12.12.2014, legit L. Brandi

(MCVE29052; GenBank KU953372); 15.01.2015 legit A. Brandi (MCVE29053;

GenBank KU953371).

Discussion

Leucoagaricus ariminensis has, as major morphological features, a rather

fleshy predominately violaceous basidioma, usually with a stipe shorter than

the pileus diameter and with a membranous purple-rimmed white annulus.

It is distinguished by spores without a germ pore and a non-gelatinous

pileus covering with somewhat elongated terminal elements that at times

taper towards the apex. Since the basidioma surfaces in L. ariminensis react

negatively to ammonia, this species would be placed in Leucoagaricus subsect.

Trichodermi Bon & Mig]. according to Bon’s (1993) classification. However, our

molecular results do not support this attribution.

Leucoagaricus ariminensis sp. nov. (Italy) ... 213

ITS sequence analyses support L. ariminensis in Leucoagaricus sect. Piloselli

(Kihner) Singer, indicating that a negative reaction to ammonia is not a

constant within this section.

In both Bayesian (Fic. 1) and ML trees, L. ariminensis is distinct from

L. ionidicolor Bellu & Lanzoni from the Netherlands (AY176415) and the

Czech Republic (KU953373). Statistics support this separation (BPP = 1,

MLB = 100%), and sequences from both L. ionidicolor collections differ from

L. ariminensis by at least 42 nucleotides.

The ITS sequence from the Czech L. ionidicolor basidioma differs from

that of the Dutch basidioma by only 5 nucleotides. The Czech specimen has

“terminal cells which are narrowly clavate when young, then very long, 80-200

x 15-20 um narrowly fusiform with rostrate apex” (Holec 2009), unlike the

pileipellis of L. ariminensis where no rostrate elements are present.

Compared to L. ariminensis, L. ionidicolor has a more slender stem, a

minutely squamulose pileus surface, and shorter basidia, (14-21 x 7-8 um,

Bellu & Lanzoni 1988; 14-21 x 5.5-6.5 um, Vellinga 2001), smaller, ellipsoid to

oblong basidiospores ((5—)5.5-6.5 x (3-)3.3-3.7(-4) um, Bellu & Lanzoni 1988;

6.0-7.0 x (3.5-)4.0-4.5 um, Vellinga 2001), and smaller clavate cheilocystidia

(23-45 x 8-11(-12) um, Bellu & Lanzoni 1988; 23-40 x 8.0-10 um often with

curved pedicel, Vellinga 2001). By contrast, the L. ariminensis spores range

from ellipsoid to oblong with a rounded apex, are rarely amygdaliform, and

show a great variability with regard to their dimension and Q within different

basidiomata.

Leucoagaricus ionidicolor var. major J. Charb. et al., which differs mainly

from the type species by its larger pileus and deeper shade of violet / purple, is

very similar to L. ariminensis from which it can be distinguished by its slightly

smaller spores (6-7 x 4—4.5(5) um), shorter (100(—150) um) fusiform terminal

pileipellis elements, and smaller and more uniformly shaped cheilocystidia

(Bon & Charbonnel 2000; no basidium dimensions provided). Further

molecular studies are needed to clarify the position of this variety within the

L. ionidicolor group.

Bon (1993) placed two more violaceous-coloured species in Leucoagaricus

subsect. Trichodermi: L. ianthinophaeus Locq. and L. ianthinosquamulosus

Guinb., both described from France.

Leucoagaricus ianthinophaeus, still not well-known possibly because of its

rarity, can be distinguished from L. ariminensis by its slender white glabrous

stipe with no trace of an annulus, smaller spores, inconspicuous cheilocystidia,

and its pileus covering composed of much shorter hyphae with terminal

214 ... Dovana & al.

elements 30-50 x 5-10 um according to Bon (1993), who re-examined

Locquin’s original material. Leucoagaricus ianthinosquamulosus can easily be

distinguished by its smaller basidiomata, definitely larger subfusiform spores

(6.5-10(-12.5) x 4-5(-6) um), shorter (16-50 um) and more variably shaped

cheilocystidia, and encrusting pigments in the pileus covering (Bon 1993,

Guinberteau 1993).

Leucoagaricus idae-fragum Guinb. et al., a pink European species

synonymized with Lepiota decorata based solely on morphology by Vellinga

(2006), differs mainly in its remarkable raspberry red cap colours, the presence

of a persistent general veil, a green ammonia reaction on gills and cap, and the

differently shaped cheilocystidia (Guinberteau et al. 1998).

Lepiota roseilivida Murrill from western North America (= Leucoagaricus

marriageae (D.A. Reid) Bon from Europe) can be diagnosed immediately

by its slender appearance, spores 6.7-9.8 x 3.8-5.7 um that are oblong or

amygdaliform in profile, and green-staining reaction of the external basidioma

surfaces to ammonia (Vellinga 2006).

Leucoagaricus brunneolilacinus Babos is a brownish lilac species with a

negative ammonium reaction. It becomes distinctly orange when bruised

and differs mainly from L. ariminensis by smaller spores (4—)4.2-5.1(-5.4) x

2.3-3.2(-3.5) um, shorter (13-26 Um) cylindrical to clavate cheilocystidia, and

a pileipellis with shorter (6-60 x 5-12(-16) tm) clavate terminal elements that

in young basidiocarps are intermixed with scattered globose cells (Candusso &

Lanzoni 1990).

Two other Leucoagaricus species with lilac basidiomes, L. purpureolilacinus

Huijsman from Europe and L. subpurpureolilacinus Z.W. Ge & Zhu L. Yang

from Asia, are phylogenetically well separated from /Piloselli and differ from

L. ariminensis mainly by their clavate cheilocystidia crowned with crystals and

different pileipellis (Vellinga 2001, Ge et al. 2015).

Acknowledgements

The authors are very grateful to Dr. Else C. Vellinga for helpful comments on

an earlier draft of the manuscript. We also thank Dr. Irmgard Krisai-Greilhuber

and Dr. Zai-Wei Ge for their pre-submission reviews and Dr. Shaun Pennycook

for nomenclatural review. We thank Dr. Jan Holec for providing the collection of

Leucoagaricus ionidicolor from the Czech Republic. We also thank Rosemary

Worrall for translating our manuscript into English and Maria Tullii providing the

microscopic line drawings.

Leucoagaricus ariminensis sp. nov. (Italy) ... 215

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MY COTAXON

January-March 2017— Volume 132, pp. 217-222

http://dx.doi.org/10.5248/132.217

Geastrum reinkingii reconsidered

LARISSA TRIERVEILER-PEREIRA & ROSA MARA BORGES DA SILVEIRA

Programa de Pés-Graduacdo em Botanica, Departamento de Botanica,

Universidade Federal do Rio Grande do Sul

Av. Bento Goncalves 9500, CEP 91501-970 Porto Alegre, RS, Brazil

" CORRESPONDENCE TO: Lt_pereira@yahoo.com. br

ABsTRACT—Morphological studies based on type specimens revealed that G. reinkingii

(originally described from Honduras, but subsequently synonymized under G. Iloydianum)

is a good species that is probably well distributed in tropical and subtropical forests in

America. Here we present a modern description and color photographs of the species, and

a comparison with the morphologically closest South American species: G. glaucescens, G.

hariotii, G. lloydianum, and G. setiferum.

KEY worps—Agaricomycetes, earthstar fungi, gasteromycetes, Geastraceae, Neotropical

mycota

Introduction

Geastrum Pers. is a widespread genus with numerous taxonomical problems.

Many authentic species have been treated as synonyms (Trierveiler-Pereira &

Silveira 2012, Zamora et al. 2014); and sometimes different species have been

identified under a single name (e.g., in G. fimbriatum Fr.) due to a lack of

differentiating morphological characters (Kasuya et al. 2012).

Several Geastrum species collected in tropical and subtropical areas of

America were described by C.G. Lloyd, who during many years received

specimens from contributors. The type specimen of G. reinkingii was collected

in 1923 in Honduras (Central America). Its original publication (Lloyd

1924) does not include a full description of the specimen, but instead, briefly

differentiates it from G. hariotii Lloyd. Later, the name also appeared, with no

additional information, in Rick’s posthumous list of the gasteroid species found

218 ... Trierveiler-Pereira & Silveira

in southern Brazil (Rick 1961). Finally, the species was considered a synonym

of G. lloydianum Rick by Ponce de Leén (1968) in his revision of the family

Geastraceae.

Recent morphological studies based on Geastrum type specimens revealed

that G. reinkingii is distinct from G. lloydianum and should be considered an

authentic species.

Materials & methods

In order to better circumscribe the species, recently collected specimens and

vouchers from herbaria BPI, ICN, LPS, PACA (Fungi Rickiani), and URM (Thiers 2016)

were examined macro- and microscopically. Macroscopic characters were described

based on observations of fresh and dried material, according to Sunhede (1989). Colors

and measurements refer to dry material unless otherwise stated. Colors of macroscopic

structures were determined according to Kornerup & Wanscher (1978). Colors of

microscopic features were defined in KOH preparation.

Observations of microscopic characters were made under a light microscope on

glass slide mounts (in 5% KOH) prepared from dried specimens. Scanning electronic

microscopy (SEM) was conducted at ‘Centro de Microscopia Eletrénica, UFRGS.

Basidiospores were coated with 15 nm of gold using a Bal-Tec SCD050 sputter coater

and photographed with a JEOL JSM-5800 scanning electron microscope.

Taxonomy

Geastrum reinkingii Lloyd, Mycol. Writings 7: 1280. 1924 [as “Geaster”]. PL, 4

BASIDIOMATA 2.0-3.4 cm high when expanded. ExopERIDIUM non-

hygroscopic, 1.4-2.7 cm high x 2.8-4.2 cm broad, split into 6-8 rays, some

ramifying at the tip, arched; mycelial layer pale yellow (1A3), usually peeling off

at maturity, if persistent, encrusted with debris; fleshy layer usually not persistent,

pale orange (5A3) to brownish orange (5C3) when fresh, then yellowish brown

(5D5) to brown (6E5) when dry; fibrous layer brownish orange (5C5) to light

brown (5D4). ENDOPERIDIUM subglobose to globose when recently exposed,

then depressed globose to ellipsoid, 0.5-1.2 cm high (not including the

peristome) x 1.4-2.0 cm broad, yellowish brown (5E4) to grayish brown (5E3,

6E3), when fresh, then yellowish brown (5D4) to dark brown (6F3) when dry,

stalked; stalk short, up to 1.0 mm high, cylindrical or flattened, dark blond

(5D4) to yellowish brown (5D5), endoperidial surface asperulate to rough due

to the presence of conical fascicules of hyphae (most easily observed at the base

of the endoperidium, near the apophysis); apophysis beige (4B3) to yellowish

brown (5D5); peristome deeply sulcate, usually strongly delimited, up to 0.45

cm high, concolor or slightly darker than endoperidium, almost black in some

specimens. GLEBA pulverulent at maturity, dark brown (6F4).

Geastrum reinkingii reconsidered ... 219

PLATE 1. Geastrum reinkingii. A. Holotype (BPI 705664). B. Detail of the endoperidium (holotype).

C. Fresh basidiome (ICN 177057). D. Dried specimens (URM 82017). E. Basidiospore (from

the holotype). F. Capillitial hyphae (from the holotype). Scale bars: A, C, D = 2 cm; B = 1 cm;

E=1 um; F=2.5 um.

CAPILLITIAL HYPHAE <8 um wide, pale yellow, brown to golden brown,

straight, not branched, thick-walled with narrow lumen, tips round, with or

without encrusted material. Bastp1ospores globose, 3.5-4.5 um in diam.

(including ornamentation), brown to dark brown, ornamented with short

220 ... Trierveiler-Pereira & Silveira

columns, some confluent; basidia not seen. HYPHAE OF THE MYCELIAL LAYER

<3 um wide, hyaline to pale yellowish, thick-walled; hyphae of the fibrous layer

3-11 um wide, yellowish to pale brown, thick-walled to solid; HYPHAE OF THE

PSEUDOPARENCHYMATOUS LAYER globose to ellipsoid, <74 um in the largest

dimension, hyaline to yellowish; HYPHAE OF THE ENDOPERIDIUM <10 um wide,

yellowish to pale brown, thick-walled to solid, the ones forming the hyphal

fascicules of the surface are tortuous, tightly interwoven with other hyphal

elements (mycosclerids).

HABITAT—growing solitary or in small groups (four or five basidiomata) on

forest soil (litter), without forming a subiculum.

DIsTRIBUTION—known from Honduras and tropical and subtropical areas

of Brazil. It is very likely that the species occurs throughout the Neotropical

rainforests and warmer regions of temperate America.

SPECIMENS EXAMINED—HONDURAS. ATLANTIDA, Tela, 20.III.1923, leg. O.A.

Reinking (BPI 705664, holotype!). BRAZIL. PERNAMBUCO, Sao Vicente Férrer, Mata

do Estado, 18.1X.2008, leg. L. Trierveiler-Pereira & Baltazar LTP180 (URM 82017, as

G. lloydianum; ICN 177060); R10 GRANDE DO SUL, Sao Leopoldo, leg. J. Rick (BPI

705665); 1933, leg. J. Rick (PACA 15976, as G. berkeleyi); Cagapava do Sul, Parque

Municipal Pedra do Segredo, 6.VI.2011, leg. L. Trierveiler-Pereira LTP277 (ICN

175626); PARANA, Foz do Iguacu, Parque Nacional do Iguacu, Trilha das Bananeiras,

9.11.2011, leg. L. Trierveiler-Pereira LTP179 (ICN 177057).

ADDITIONAL SPECIMENS EXAMINED— Geastrum glaucescens: ARGENTINA. BUENOS

AIRES, La Plata, IX.1894, leg. C. Spegazzini (LPS 15860, holotype).

Geastrum hariotii: BRAZIL. C.G. Lloyd’s Collection cat. No. 52535 (BPI 841469,

holotype).

Geastrum lloydianum: BRAZIL. Leg. J. Rick, C.G. Lloyd’s Collection cat. No.

57279 (BPI 841471, holotype).

Geastrum setiferum: BRAZIL. PERNAMBUCO, Serra Negra, 28.V.2002, leg.

I.G. Baseia (URM 77077, paratype); PARAiBA, Mataraca, Mineradora Millenium

(Cristal), mata controle, 2.[X.2009, leg. L. Trierveiler-Pereira (URM 82118).

COMMENTS—Geastrum reinkingii is characterized by medium-sized basidioma,

arched exoperidium; usually not persistent mycelial and fleshy layer; subglobose

to ellipsoid, stalked endoperidium, with dark, asperulate to rough endoperidial

surface, and a whitish apophysis.

The endoperidial stalk and the apophysis are not clearly evident when

the endoperidium is recently exposed, but both are remarkable features in

dried collections. Some herbarium specimens exhibit a whitish pruina on

the endoperidial surface. The basidiospores are typically dark brown and

with discrete ornamentation. Immature forms were not found in herbarium

collections or in the field, therefore basidia morphology remains unknown.

Brazilian specimens show no major differences from the holotype, the whitish

Geastrum reinkingii reconsidered ... 221

apophysis and short-stalked blackish depressed globose endoperidium being

the most remarkable features. Basidiospore measurements and morphology are

constant among the examined specimens.

One G. reinkingii collection from Southern Brazil, sent by Rick to Lloyd, is at

BPI (705665), but no collections were deposited at PACA, Rick’s most important

fungal collection. However, one of Rick’s collections identified as G. berkeleyi

(PACA 15976) does correspond to G. reinkingii. One G. reinkingii specimen

(URM 82017) was reported from Northeastern Brazil as G. lloydianum

(Trierveiler-Pereira et al. 2011).

Geastrum hariotii and G. lloydianum are very closely related species, and

their differences have been discussed by Trierveiler-Pereira & Silveira (2012).

Microscopically, the differences separating G. reinkingii and G. lloydianum are

evident, since the G. lloydianum basidiospores are 5-6 um in diam., yellowish

brown, and prominently ornamented. Since there are a limited number of

morphological characters available to differentiate species of Geastrum, we still

believe that basidiospore morphology is an important diagnostic feature. The

G. hariotii basidiospores are more similar to those of G. reinkingii; nonetheless,

the two species are macroscopically distinct: G. hariotii produces smaller

basidiomata with a globose sessile, not asperulate, endoperidium. Moreover,

no whitish apophysis is observed in G. hariotii.

Geastrum setiferum Baseia produces large arched basidiomata and a short

stalked endoperidium (Baseia & Milanez 2002, Trierveiler-Pereira et al. 2011).

In many aspects it resembles G. reinkingii, but the presence of dark setae in the

G. setiferum endoperidium is an important distinguishing feature. Moreover,

the endoperidial surface of G. setiferum is lighter (usually light brown), not

remarkably ellipsoidal, without a whitish apophysis, and the peristome

morphology is a transition between fibrillose and plicate.

Geastrum glaucescens Speg. was described from Argentina and its basidiomes

are small, arched, and dark colored. The endoperidium is globose, brownish

to blackish, and covered with abundant persistent white mesoperidial matter

(Kuhar et al. 2013). According to the protologue, the endoperidium is short

stalked (Spegazzini 1912), but this feature was difficult to confirm by analysis of

the type specimen, which is sectioned in half. Soto & Wright (2000) illustrated

the species as clearly stalked. The primary macroscopical features distinguish

this species from G. reinkingii, as depicted by Kuhar et al. (2012).

Acknowledgments

The senior author thanks Jean Louis Pierre for discussions on the taxonomy of

Geastrum; curators and staff of all cited herbaria for the access to the exsiccatae;

222 ... Trierveiler-Pereira & Silveira

and colleagues that helped during fieldwork. PROPG-UFRGS (‘Edital 001/2013’)

and PROTAX (‘Edital MCT/CNPq/MEC/CAPES 52/2010’) are acknowledged

for financial support. This study is a partial result of the Ph.D. thesis by LTP, with a

scholarship provided by the Brazilian government (agency: CAPES). We also thank the

contributions of the pre-submission reviewers, Dra. Maria Luciana Hernandez Caffot

(Universidad Nacional de Jujuy-CONICET, Argentina) and Dra. Tiara Sousa Cabral

(Instituto Nacional de Pesquisas da Amazénia-INPA, Brazil).

Literature cited

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endoperidium. Mycotaxon 84: 135-140.

Kasuya T, Hosaka K, Uno K, Kakishima M. 2012. Phylogenetic placement of Geastrum

melanocephalum and polyphyly of Geastrum triplex. Mycoscience 53(6): 411-426.

http://dx.doi.org/10.1007/s10267-012-0186-z

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

243 p.

Kuhar F, Castiglia V, Papinutti L. 2012. Geastrum species of the La Rioja province, Argentina.

Mycotaxon 122: 145-156. http://dx.doi.org/10.5248/122.145

Lloyd GC. 1924. Mycological Notes N° 72. Mycological Writings 7: 1269-1300.

Ponce de Leon, P. 1968. A revision of the Geastraceae. Fieldiana, Bot. 31: 303-349.

Rick J. 1961. Basidiomycetes Eubasidii no Rio Grande do Sul - Brasilia. Iheringia 9: 451-480.

Soto MK, Wright JE. 2000. Taxonomia del género Geastrum (Basidiomycetes, Lycoperdales) en la

Provincia de Buenos Aires, Argentina. Bol. Soc. Argent. Bot. 34(3-4): 185-201.

Spegazzini C. 1912. Mycetes Argentinenses. Series VI. Anales Mus. Nac. Buenos Aires 23: 1-146.

Sunhede S. 1989. Geastraceae (Basidiomycotina). Morphology, ecology, and systematics with

special emphasis on the North European species. Syn. Fungorum 1: 1-534.

Thiers B. 2016. Index Herbariorum: a global directory of public herbaria and associated staff. New

York Botanical Garden's Virtual Herbarium. Available from: http://sweetgum.nybg.org/ih/

(accessed: June 2016).

Trierveiler-Pereira L, Calonge FD, Baseia IG. 2011. New distributional data on

Geastrum (Geastraceae, Basidiomycota) from Brazil. Acta Bot. Bras. 25(3): 577-585.

http://dx.doi.org/10.1590/S0102-3306201 1000300010

Trierveiler-Pereira T, Silveira RMB. 2012. On the Geastrum species (Geastraceae, Basidiomycota)

described by Rick. Phytotaxa 61: 37-46. http://dx.doi.org/10.11646/phytotaxa.61.1.3

Zamora JC, Kuhar F, Castiglia V, Papinutti L. 2014. On Geastrum argentinum, a forgotten species.

Mycoscience 55(3): 177-182. http://dx.doi.org/10.1016/j.my c.2013.08.003

MY COTAXON

January-March 2017— Volume 132, pp. 223-229

http://dx.doi.org/10.5248/132.223

Bacidia, Micarea, Sagedia, and Stigmidium species

new to Turkey

KADIR KINALIOGLU '* & ANDRE APTROOT ”

‘Department of Biology, Faculty of Science and Arts, Giresun University, Giresun, Turkey*

?Adviesbureau voor Bryologie en Lichenologie,

G.v.d. Veenstraat 107, NL-3762 XK Soest, The Netherlands

CORRESPONDENCE TO: kkinalioglu@hotmail.com & andreaptroot@gmail.com

AsBstTRAcT—During lichenological exploration in Turkey, two lichenised fungi (Micarea

micrococca, Sagedia zonata) and one lichenicolous fungus (Stigmidium microspilum) were

established as new records for Asia, and one lichenised fungus (Bacidia chloroticula) as new

for Turkey. Illustrations are accompanied by brief descriptions and comments on habitat,

substrate, and geographical distribution.

Key worps—biodiversity, Corylus, Giresun, Istanbul, Trabzon

Introduction

Studies on the lichen biodiversity of Turkey are as yet incomplete, but many

new lichenized and lichenicolous fungi records for Turkey have been found in

recent years (Kinalioglu & Aptroot 2010, Kocakaya et al. 2011, Karagéz & Aslan

2012, Aslan & Yazici 2013, Halici 2015, Yazic1 & Aptroot 2015, Senkardesler

et al. 2016). Despite the increase in the number of lichen records, additional

studies are needed in some places where data on lichen biodiversity are still

scarce. The objective of this paper is to add new species to the list of lichens

known from Turkey.

Materials & methods

The study is based on lichen material collected in three different Turkish

provinces (Giresun, Istanbul, Trabzon). Vouchers are preserved in the herbarium of

the Biology Department, Faculty of Science and Arts, Giresun University, Giresun,

Turkey (GUB).

224 ... Kinalioglu & Aptroot

Morphological, anatomical, and chemical features of the lichen thalli were

examined with a stereomicroscope, a light microscope and the standard chemical spot

tests (e.g. 10% potassium hydroxide).

Species recorded

Bacidia chloroticula (Nyl.) A.L. Sm., Monogr. Brit. Lich. 2: 155. 1911. FIG. 1

Thallus thin, grey—pale green. Photobiont cells 5-12.5 um diam. Apothecia

small, flat, 0.07-0.3 mm diam., pinkish to pale brown. Hymenium colourless,

32.5-40 um tall, epihymenium and hypothecium colourless. Ascospores

1-4-celled, 20-35 x 1-1.6 um, acicular. Thallus C-, K-, KC-, PD-.

SPECIMEN EXAMINED: TURKEY. [Stanbul, Maltepe, in the vicinity of Basibiiyiik,

40°57°32”N 29°09’27”E, 235 m, on siliceous rocks, 24 April 2007, Kinalioglu (GUB

2383).

REMARKS—For a detailed description of the species see Smith et al. (2009).

Bacidia chloroticula is a pioneer species. It grows on various shaded, often

man-made substrata, such as tree bases and exposed roots, dead grasses, tops

of cut stumps and fence posts, cement and brick rubble, and even painted

Fic. 1. Bacidia chloroticula (GUB 2383): habit. Scale bar = 1 mm.

Lichenized and lichenicolous fungi new for Turkey... 225

iron-work and tarmacadam (Smith et al. 2009). In Turkey, we have recorded it

only once, from siliceous rocks at low elevation.

Known previously from Asia, Europe, and North America (Smith et al.

2009). New to Turkey.

Bacidia chloroticula is most easily confused with Bacidina saxenii (Erichsen)

M. Hauck & V. Wirth, which has a very similar ecology. It differs in having

often darker, pale to dark-brown apothecia with pale brown hymenium and

larger, vesicle-like, 20 x 12 mm large cells in the outer exciple (Smith et al.

2009). In B. chloroticula the cells in the exciple do not exceed 5(-7) um in

width (Smith et al. 2009). The size of the apothecia and hymenium in the

Turkish collection is almost the same as given by Smith et al. (2009; apothecia

0.08-0.2 mm. diam., hymenium 40-42 um). However, the ascospores in the

Turkish material are slightly smaller than those reported by Smith et al. (2009;

30-50 x 2-3um).

Micarea micrococca (Korb.) Gams ex Coppins,

Checkl. Lich. Gr. Brit. Ire.: 86. 2002. Fic. 2

Thallus crustose, greenish. Apothecia convex to subglobose, immarginate,

mostly tuberculate, 0.1-0.3 mm diam., to 0.5 mm diam. when tuberculate,

pinkish to flesh or rarely pale brownish. Epihymenium colourless, K-,

hymenium 32.5-45 um tall, colourless. Asci clavate, 8-spored. Ascospores

ellipsoid to oblong, hyaline, 1-celled, 7.5-10 x 2.5-3 um. Thallus and apothecia

C-, K-, KC-, PD-.

SPECIMEN EXAMINED: TURKEY. Giresun: Espiye, Cepnikéy, 40°51’38”N 38°43’57”E,

493 m, on garden fence, 5 October 2013, Kinalioglu (GUB 2384).

REMARKS—For detailed descriptions of the species see Smith et al. (2009) and

Barton & Lendemer (2014).

Micarea micrococca, the most common representative of the M. prasina

group, is found on acid bark of trees and stumps, shrubs, plant debris

(usually in conifer plantations), soil or debris in rock crevices in coastal sites,

rarely on shaded sandstone rocks, mostly in shaded sites or niches, even close

to large city centres (Smith et al. 2009). In Turkey, Micarea micrococca was

collected on a garden fence next to a shady hazelnut field.

Known previously from Australia, Europe, and North America (Smith et

al. 2009). New to Asia.

Micarea micrococca is extremely variable. It is very similar to M. prasina

Fr. and can sometimes only be separated by using TLC. The apothecia are

the clearest morphological character differentiating M. micrococca from

226 ... Kinalioglu & Aptroot

Fic. 2. Micarea micrococca (GUB 2384): habit. Scale bar = 1 mm.

M. prasina, which has whitish, smaller apothecia (<0.2 mm diam.; Smith et al.

2009, Dobson 2011).

The ascospores and hymenium in the Turkish specimen are smaller than

those cited by Barton & Lendemer (2014; ascospores 7.4-10.9 x 2.3-3.6 um,

hymenium 40-60 tm) and Smith et al. (2009; ascospores 6-17 x 2-4.5 um,

hymenium 30-50 um). The Turkish collection differs from North American

samples by the K- reaction.

Sagedia zonata Ach., Kongl. Vetensk. Acad. Nya Hand. 30: 165. 1809. FIG. 3

Thallus crustose, saxicolous, greyish or very slightly bluish, irregularly

cracked-areolate, areoles to 1.5 mm wide. Apothecia black, 0.25-1 mm

diam., numerous, immersed or somewhat raised, with a prominent black

rim. Hymenium 80-95 um tall, colourless. Asci 8-spored. Ascospores simple,

mostly subglobose, broadly ellipsoid or very rarely ellipsoid, 13.5-20 x

7.5-11 um. Thallus C-, K-, KC-, PD-.

SPECIMEN EXAMINED: TURKEY. Trabzon: Arakh, S of Kizilkaya Yaylasi, 40°39’56”N

40°01’49”E, 2300 m, on siliceous rocks, 17 August 2005, Kinalioglu (GUB 2385).

REMARKS—For a detailed description of the species see Wirth et al. (2013).

This boreal-temperate species grows on non-calcareous blocks, rocks, and

boulders in slightly shady to mostly well-lit areas, but also on periodically or

Lichenized and lichenicolous fungi new for Turkey ... 227

Fic. 3. Sagedia zonata (GUB 2385): habit. Scale bar = 1 mm.

rarely inundated surfaces next to streams in montane to subalpine zones (Wirth

et al. 2013). In Turkey, we collected it from siliceous rocks in an exposed area.

Known previously from Europe (Wirth et al. 2013). New to Asia.

The Turkish material of Sagedia zonata differs from European specimens by

having bigger areoles and a lower hymenium. In the European collections the

areole is 0.5-1 mm diam., and the hymenium is 85-100 um high (Wirth et al.

2013). Otherwise the Turkish collection matches the sizes given by Wirth et al.

(2013; apothecia 0.2-1 mm diam., ascospores 14—19 x 8-12 um).

Stigmidium microspilum (K6rb.) D. Hawksw., Kew Bull. 30(1): 201.1975. FIG. 4

Thallus dark brownish circular patches, patches <0.5-1.5 mm _ wide,

regularly scattered on the thallus surface of the host (Graphis scripta (L.) Ach.).

Perithecia small, black, 0.1-0.12 mm diam. Ascospores hyaline, elongate or

narrowly ellipsoid, 2-celled, 14—18.8 x 2.5-3.8 um. Thallus C-, K-, KC-, PD-.

228 ... Kinalioglu & Aptroot

Fic. 4. Stigmidium microspilum (GUB 2386): habit. Scale bar = 1 mm.

SPECIMEN EXAMINED: TURKEY. GiRESuN: Espiye, N of Yenikdy village, 40°4’42”N

38°45'31”E, 781 m, on the thallus of Graphis scripta on Corylus sp., 6 October 2013,

Kinalioglu (GUB 2386).

REMARKS—For a detailed description of S. microspilum, see Hawksworth (1983).

This widespread lichenicolous species occurs on thalli of Graphis scripta on

Corylus sp. in old woodlands (Hawksworth 1983). In Turkey it was growing on

a Graphis scripta thallus in a damp hazelnut garden.

Known previously from Europe (Hawksworth 1983). New to Asia.

The Turkish material of Stigmidium microspilum has slightly longer

and narrower ascospores than those described from European material

(Hawksworth 1983; ascospores 14-15 x 3-5 um).

Acknowledgements

We are grateful to Dr. H. Sipman and Dr. L. Lok6s for reviewing the manuscript and

providing valuable comments.

Literature cited

Aslan A, Yazici1 K. 2013. New Lecanora, Lecidea, Melaspilea, Placynthium, and Verrucaria

records for Turkey and Asia. Mycotaxon 123: 321-326. http://dx.doi.org/10.5248/123.321

Barton J, Lendemer JC. 2014. Micarea micrococca and M. prasina, the first assessment

of two very similar species in eastern North America. Bryologist 117(3): 223-231.

http://dx.doi.org/10.1639/0007-2745-117.3.223

Lichenized and lichenicolous fungi new for Turkey ... 229

Dobson FS. 2011. Lichens: an illustrated guide to the British and Iris species. Richmond

Publishing, Slough. 496 p.

Halici MG. 2015. New records of crustose Teloschistaceae and lichenicolous fungi from Turkey.

Mycotaxon 130(3): 769-773. http://dx.doi.org/10.5248/130.769

Hawksworth DL. 1983. A key to the lichen-forming, parasitic, parasymbiotic and

saprophytic fungi occurring on lichens in the British Isles. Lichenologist 15(1): 1-44.

http://dx.doi.org/10.1017/S0024282983000031

Karag6z Y, Aslan A. 2012. Floristic lichen records from Kemaliye District (Erzincan) and Van

Province. Turkish Journal of Botany 36: 558-565. http://dx.doi.org/10.3906/bot-1101-30

Kinahioglu K, Aptroot A. 2010 Catillaria, Cladonia, Strigula, and Cresporhaphis species new to

Turkey and Asia, Mycotaxon 114: 329-332. http://dx.doi.org/10.5248/114.329

Kocakaya M, Halict MG, Aksoy A. 2011. Zwackhiomyces turcicus sp. nov. (Ascomycota,

Xanthopyreniaceae) from Turkey. Mycotaxon 116: 329-333. http://dx.doi.org/10.5248/116.329

Senkardesler A, Cansaran Duman D, Lokés L, Ahti T. 2016. Cladonia trapezuntica

(Cladoniaceae, lichenized Ascomycota): a robust morphotype of Pycnothelia papillaria,

a taxonomic study with conservational survey. Turkish Journal of Botany 40: 104-111.

http://dx.doi.org/10.3906/bot- 1403-49

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA. 2009. The

lichens of Great Britain and Ireland. British Lichen Society, London, 1046 p.

Wirth V, Hauck M, Schultz M. 2013. Die Flechten Deutschlands (2 volumes). E. Ulmer, Stuttgart.

1244 p.

Yazici K, Aptroot A. 2015. Buellia, Lempholemma, and Thelidium species new for Turkey and Asia.

Mycotaxon 130(3): 701-706. http://dx.doi.org/10.5248/130.701

MY COTAXON

January-March 2017— Volume 132, pp. 231-234

http://dx.doi.org/10.5248/132.231

Urocystis narcissi, a new record from Asia

SADIQULLAH *, A. ISHAQ 2, M. Fiaz’, A.N. KHALID 7 & H. AHMAD 3

"Department of Botany & * Department of Genetics,Hazara University,

Mansehra, Dhodial 21310, Pakistan

? Department of Botany, University of the Punjab,

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

* CORRESPONDENCE TO: muhammadfiazhu@gmail.com

AxBsTRACT—Urocystis narcissi, reported here from Pakistan, represents a new record for

Asia. This smut fungus was collected for the first time outside its type locality in Spain.

Narcissus tazetta is recognized as a new host plant for this rare smut.

Key worps—Shangla, Swat, taxonomy, Ustilaginales, Ustilaginomycetes

Introduction

During a survey of Ustilaginales in the Shangla and Swat districts of Khyber

Pakhtunkhwa (KP) Province, Pakistan, Narcissus tazetta was found infected

with Urocystis narcissi. This rare smut fungus was previously known only from

its type locality in Spain. The sampling sites are neighborhood districts located

at 34°34’-35°55’N 71°08’-72°50’E. Narcissus tazetta was found infected with

smut in two areas of Khyber-Pakhtunkhwa (Ahmad & Sirajuddin 1996): one

infected sample was collected from Kabal valley, Swat, in February 2015, and

the second from Puran valley, Shangla, in March 2015. Nine smut fungi have

previously been reported from Swat, but none from Shangla (Ahmad et al.

1997).

Materials & methods

After collection of the infected plants with their inflorescences, the specimens were

photographed, and preserved. Free hand sections were cut and scrape mounts of the

infected portions were observed under a Labomed CSM2 stereomicroscope. For light

microscopic (LM) observations, spores were mounted in lacto phenol solution on

glass slides, gently heated to boiling point to rehydrate the spores, and then cooled.

232 ... Sadiqullah & al.

Preparations were examined using a Nikon YS 100 microscope, and photographed

with the aid of a Digipro-labomed. Forty spores were measured using a Zeiss

eyepiece screw micrometer. For SEM, the spores were critically point dried, attached

to specimen holders with double sided adhesive tape, and coated with a 50 nm gold

film in a Polaron E5300 freeze drier. The gold-coated specimens were observed and

photographed in a Camscan 3-30BM scanning electron microscope. Specimens have

been deposited in the herbaria at the University of the Punjab, Lahore, Pakistan (LAH)

and Hazara University Mansehra, Pakistan (HUP).

—

Fic.1: Urocystis narcissi. A, B, D. Infected host plants. C, E. Spore balls in LM.

Scale bars: A, B = 51 um; C, D=10 um; E=11 um; F= 8 um.

Urocystis narcissi new to Pakistan ... 233

Taxonomy

Urocystis narcissi (Gonz. Frag.) Vanky, Mycotaxon 48: 41 (1993) FIGS 1, 2

= Urocystis colchici f. narcissi Gonz. Frag., Trab. Mus.

Nac. Cienc. Nat., Ser. Bot. 7: 22 (1914)

= Turburcinia narcissi (Gonz. Frag.) Cif., Fl. Ital. Crypt., Fungi 1(17): 101 (1938)

Sori in leaves and scales, yellowish-silver or yellowish-brown or black

pustules containing a black powdery mass. Spore balls globose to subglobose

or ellipsoid, 25-44 x 31-54 um, composed of 1-3(-4) spores, completely

surrounded by sterile cells. Spores globose to subglobose, ovate or ellipsoidal,

12-21 x 13-24 um, dark brown, walls smooth; sterile cells variable in shape and

size, subglobose to elongated, 4-14 um long, wall 0.6-1.5 um thick, yellowish

brown to hyaline, smooth.

SPECIMENS EXAMINED: On Narcissus tazetta L. (Amaryllidaceae): PAKISTAN, KHYBER

PAKHTUNKHWA PROVINCE, Swat District, Kabal valley Deolai, 1000 m a.s.1 , February

2015, leg. Sadiqullah SUS-02 (HUP 315); Shangla District, Puran Aloch, 1100 ma.s.l.,

March 2015, leg. Sadiqullah SUS-03 (HUP 316).

ComMMENTS—Urocystis narcissi has previously been reported on Narcissus

pallidulus Graells from the Sierra de Guadarrama, Spain (Gonzalez Fragoso

1914, as U. colchici f. narcissi; Ciferri 1938, as Turburcinia narcissi; Vanky 1993,

2011). Our collections on N. tazetta from Pakistan are similar to those cited in

x4,3898 Sum CRL UOP

X1,986 16xm CRL UOP Sum CRL UOP

Fic. 2: Urocystis narcissi. Spore balls in SEM.

234 ... Sadiqullah & al.

Vanky (2011), except that the maximum spore size in our material was slightly

larger. Narcissus tazetta is a new host record for this fungus.

Acknowledgments

We sincerely thank Dr. Kalman Vanky, Honorary Member of Hungarian

Academy of Science, for verification of the fungus and Mr. Mehboob Ur Rahman for

verification of the host plant. We are highly obliged to Prof. Dr. Cvetomir M. Denchev

(Bulgarian Academy of Sciences, Sofia) and Dr. Najam-ul-Sehar Afshan (University

of the Punjab, Lahore) for acting as pre-submission reviewers.

Literature cited

Ahmad H, Sirajuddin. 1996. Ethnobotanical profile of Swat. 202-206, in: ZK Shinwari et al. (eds).

Proceedings of First Training Workshop on Ethnobotany and its Application to Conservation.

National Herbarium/PARC, Islamabad.

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

Pakistan, Department of Botany, University of the Punjab, Quaid-e-Azam campus, Lahore.

Ciferri R. 1938. Ustilaginales: Tilletiaceae, Graphiolaceae, Ustilaginaceae. Flora Italica Cryptogama,

Pars I: Fungi, Fasc. 17. 443 p.

Gonzalez Fragoso R. 1914. Nueva contribucién a la flora micolédgica del Guadarrama:

Teleomicetos y Deuteromicetos (adiciones). Trabajos del Museo Nacional de Ciencias

Naturales, Serie Botanica, Num. 7. 80 p.

Vanky K. 1993. Taxonomical studies on Ustilaginales. X. Mycotaxon. 48: 27-44

Vanky K. 2011 [“2012”]. Smut fungi of the world. APS Press, St. Paul, Minnesota, USA. xvii +

1458 p.

MY COTAXON

January-March 2017— Volume 132, pp. 235-240

http://dx.doi.org/10.5248/132.235

Lembosia dianesei sp. nov. associated with

Peritassa campestris in Minas Gerais, Brazil

MARUZANETE PEREIRA MELO’, JOSE LUIZ BEZERRA’ ,

SILVINO INTRA MOREIRA? & EDUARDO ALVES?

"Departamento de Fitotecnia, Universidade Federal do Piaui,

Campus UFPI, Av. Universitaria, Ininga, Teresina PI, 64049-550, Brazil

?Universidade Federal do Recéncavo da Bahia, Centro de Ciéncias Agrarias, Ambientais e Bioldgicas,

Campus UFRB, Centro, Cruz das Almas BA, 44380-000, Brazil

°Universidade Federal de Lavras, Departamento de Fitopatologia,

Campus UFLA, Centro, Lavras MG, 37200-000, Brazil

“CORRESPONDENCE TO: jlulabezerra@hotmail.com

ABSTRACT—A new Lembosia species is described in association with Peritassa campestris

from a cerrado region of Minas Gerais, Brazil. The new species is described based on

morphological characteristics and compared with other Lembosia species associated with

celastraceous hosts.

Key worps— Ascomycota, Asterinaceae, Celastraceae, cerrado

Introduction

The plant genus Peritassa Miers (Celastraceae R. Br.) includes nine species

endemic to Brazil. These species are distributed across many Brazilian states and

are found in the cerrado, cerradao, and campo ralo biomes. Peritassa campestris

has been used pharmacologically, and the local population consumes the fruit

in natura. However, since 2009 the leaves of some P campestris plants in the

Cristalia and Botumirim municipalities of Minas Gerais have been found to

have a black colored fungal colonization. This infection occurs most frequently

during the rainy season. The fungal colonies have macroscopic characteristics

similar to those of Meliolales Gaum. ex D. Hawksw. & O.E. Erikss. However,

laboratory observations revealed that the fungus represents a species of

Lembosia Lév., (Asterinaceae Hansf.). This genus is marked by the presence

236 ... Melo & al.

of hyphopodiated mycelium, linear hysteriothecia with radiate upper walls,

parallel bitunicate asci, and light brown bicellular ascospores (Bezerra 2004,

Hofmann & Piepenbring 2014, Song & Hosagoudar 2003). Additionally,

the identity of the host plant is required to distinguish Lembosia species

(Inacio & Cannon 2003, Miller & von Arx 1962, Mebey & Hawksworth

1995). Asterinaceae are usually associated with tropical and subtropical plants

around the globe (Hofmann & Piepenbring 2014, Guatimosim et al. 2015).

Infection has been reported in wild plants but also occurs in cultivated plants,

such as Asterina manihotis Syd. on living leaves on Manihot esculenta Crantz

(Hofmann & Piepenbring 2008). New species of Asterinaceae have been

described from Brazil, including Lembosia bezerrae Firmino & O.L. Pereira

and L. epidendri Meir. Silva & O.L. Pereira (Silva & Pereira 2008, Firmino

& Pereira 2014). Here we propose another new species, Lembosia dianesei,

described and illustrated here.

Materials & methods

Peritassa campestris leaves were collected and sent to the Electron Microscopy

and Ultrastructural Analysis Laboratory at the Universidade Federal de Lavras

(LME/UFLA). Fragments and manual sections were mounted on slides in

lactoglycerol for light microscopy examination of fungal structures. Melzer’s

reagent was used to observe the amyloid zones of the asci. Permanent slides were

made using PVLG (Polyvinyl alcohol-lactic acid-glycerol). Observations were made

with a Leica DM2000 bright field vertical microscope using LAS v.4.5 software.

The leaf fragments used for scanning electron microscopy studies were fixed in

Karnovsky solution (pH 7.2), post-fixed with 1.0% osmium tetroxide solution, and

dehydrated in an acetone series (Bozzola & Russell 1999). The specimens were dried

in a Bal-tec CPD 030 critical point dryer and coated with gold in a Bal-tec SCD 050

sputter coater for observation using the LEO EVO 40 and Smart Sem softwareat20

Kv scanning electron microscope with a working distance of 9 mm. Both light and

scanning electron micrographs were edited in Corel Draw X7.

Two representative specimens were deposited in the herbarium of the Universidade

Federal de Vicosa, Minas Gerais, Brazil (VIC).

Taxonomy

Lembosia dianesei M.P. Melo, J.L. Bezerra, S.I. Moreira & E. Alves, sp. nov. PL. 1

MycoBAnk MB 814528

Differs from Lembosia albersii by its smaller asci and its supra-median ascospore septum;

and from L. lophopetali and L. salaciae by its smaller asci and its shorter unicellular

hyphopodia.

Type: Brazil, Minas Gerais, Cristalia, on living leaves of Peritassa campestris (Cambess.)

A.C. Sm. (Celastraceae), 30 July 2015, M.P. Melo (Holotype, VIC 42934).

Lembosia dianesei sp. nov. (Brazil) ... 237

PLaTE 1. Lembosia dianesei (VIC 42934) on Peritassa campestris. A: hysteriothecia and mycelium

(SEM). B: epiphyllous colonies. C, G: asci and ascospores. D, E: section through ascoma.

F: uniseptate ascospores, constricted at the septum. Scale bars: A, F = 10 um; C-E, G = 40 um.

238 ... Melo & al.

ErymMo oey: In honor of the Brazilian mycologist, José Carmine Dianese.

Cotonies epiphyllous, irregular or circular, single or confluent, black color,

hyphae straight flexuous, branched at 45° angle, brown, septate, hyphal cells

cylindrical 3.4-5.3 um diam., smooth. HypHopPopia (appressoria) numerous,

lateral, alternated and unilaterally distributed, nodule-shaped, unicellular,

4.5-7.1 x 4.7-6.6 um, brown, penetration peg central on the appressorial cell.

No haustoria were observed. ASCoMATA superficial, hysteriothecia, single to

confluent, fringed at margins, linear or Y-X shaped, 150-335 x 130-160 um,

with a height of 29-70.5 um, opening by a longitudinal dehiscence, dark brown;

upper wall of textura prismatica, cells radiating, rectangular, 6.5-13.2 um thick,

brown; basal wall 3.5-16.3 um thick. ASCI bitunicate, disposed as an upright

palisade layer, ellipsoid to subglobose, 8-spored, hyaline to brown, 28.7-36.3

x 14.4-19.2 um. Ascospores oblong to ellipsoid, ends rounded, straight,

1-septate, slightly constricted at the supra-median septum, hyaline when young

and brown at maturity, smooth, conglobed, 19.7-28.7 x 6.8-10 um; apical cell

smaller, basal cell larger and thinner. Asexual morph not seen.

ADDITIONAL SPECIMEN EXAMINED: BRAZIL, MINAs GERAIS, Botumirim, on living

leaves of Peritassa campestris, 30 July 2015, M.P. Melo (VIC 42935).

Comments

Three Lembosia species have been previously reported in association

with living leaves of celastraceous hosts: L. salaciae Hosag. & Archana,

associated with Salacia sp. from India (Hosagoudar & Archana 2009);

L. albersii Henn., associated with Elaeodendron sp. from Tanzania (Hennings

1902); and L. lophopetali (Rehm) Hansf., associated with Lophopetalum

toxicum from the Philippines (Hansford 1949). Three additional records of

Lembosia spp. have been reported from Brazil associated with living leaves

of undetermined (or unstated) celastraceous hosts (Farr & Rossman 2015).

The three previously described species are easily distinguished from

Lembosia dianesei (TABLE 1): L. albersii differs by its larger asci and its

median ascospore septum (Hennings 1902); L. lophopetali differs by its

longer two-celled hyphopodia, its larger asci, and its wider ascospores

(Hansford 1949); and L. salaciae differs by its longer two-celled hyphopodia

and its larger ascomata, asci, and ascospores (Hosagoudar & Archana 2009).

Another peculiar characteristic of Lembosia dianesei is its strong hyphal

adherence to the foliar surface.

Acknowledgements

The authors thank Jaime Pereira de Melo, a technician at the Empresa de Extensao

Rural do Estado de Minas Gerais (EMATER-MG), for his support in collecting

Lembosia dianesei sp. nov. (Brazil) ... 239

TABLE 1. Comparisons of hosts and morphology in Lembosia species

associated with Celastraceae

HyYPHOPODIA ASCOMATA ASCI ASCOSPORES

SPECIES Host

(um) (wm) (wm) (wm)

L. dianesei Peritassa 4.5-7.1 205-335 28.7-36.3 19.7-28.7

campestris x 130-160 x 14.4-19.2 x 6.8-10

L. salaciae Salacia sp. 9-18 500-560 32-42 28-35

x 250-300 x 26-35 x 12-18

L. albersii Elaeodendron sp. — — 45-90 20-30

x 20-30 x 10-12

L. lophopetali Lophopetalum — — 40-50 25-30

toxicum x 20-25 x 11-14

specimens, Capes and CNPq for the scholarship given to J.L. Bezerra, and the

Fundacao de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG) for financial

support. They are also grateful to Jadergudson Pereira (Universidade Estadual de

Santa Cruz, Brazil) and Olinto Liparini Pereira (Universidade Federal de Vicosa,

Brazil) for helpful comments and presubmission review.

Literature cited

Bezerra JL. 2004. Taxonomia de ascomicetos: revisao da ordem Asterinales. Revisio Anual de

Patologia de Plantas 12: 91-115.

Bozzola JJ, Russell LD (eds). 1999. Electron microscopy: principles and techniques for biologists,

24 edition. Jones and Bartlett Publishers, USA.

Farr DF, Rossman AY. 2014. Fungal database, Systematic Mycology and Microbiology Laboratory,

ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/ (viewed online on November 14, 2014).

Firmino AL, Pereira OL. 2014. Lembosia bezerrae, a new asterinaceous fungus

associated with a terrestrial orchid from Bahia, Brazil. Mycotaxon 127: 199-205.

http://dx.doi.org/10.5248/127.199

Guatimosim E, Firmino AL, Bezerra JL, Pereira OL, Barreto RW, Crous PW. 2015. Towards a

phylogenetic reappraisal of Parmulariaceae and Asterinaceae (Dothideomycetes). Persoonia

35: 230-241. http://dx.doi.org/10.3767/003158515x688046

Hansford CG. 1949. Tropical fungi.—III. New species and revisions. Proceedings of the Linnean

Society of London 160(2): 116-153. http://dx.doi-org/10.1111/j.1095-8312.1949.tb00521.x

Hennings PC. 1902. Fungi africae orientalis. IH. Botanische Jahrbiicher fiir Systematik

Pflanzengeschichte und Pflanzengeographie. 33: 34-40.

Hofmann TA, Piepenbring M. 2008. New species and records of Asterina from Panama.

Mycological Progress 7: 87-98. http://dx.doi.org/10.1007/s11557-008-0555-3

Hofmann TA, Piepenbring M. 2014. New records of plant parasitic Asterinaceae (Dothideomycetes,

Ascomycota) with intercalary appressoria from Central America and Panama. Tropical Plant

Pathology 39: 419-427. http://dx.doi.org/10.1590/S1982-56762014000600001

Hosagoudar VB, Archana GR. 2009. Studies on foliicolous fungi-X XVII. Indian Journal of Science

& Technology 2: 1-26.

240 ... Melo & al.

Inacio CA, Cannon PF. 2003. Viegasella and Mintera, two new genera of Parmulariaceae

(Ascomycota) with notes on the species referred to Schneepia. Mycological Research 107: 82-92.

http://dx.doi.org/10.1017/s0953756202007013

Mebey RK, Hawksworth DL. 1995. Diporothecaceae, a new family of ascomycetes, and the term

“hyphopodium”. Systema Ascomycetum 14: 25-31.

Mendes MAS, da Silva VL, Dianese JC (eds). 1998. Fungos em plantas no Brasil. Embrapa-SPI/

Embrapa-Cenargen, Brasil.

Miller E, von Arx JA. 1962. Die Gattungen der didymosporen Pyrenomyceten. Beitrage zur

Kryptogamenflora der Schweiz. 11(2). 922 p.

Silva M, Pereira OL. 2008. Black mildew disease of neotropical orchid Epidendrum secundum

caused by Lembosia epidendri sp. from Minas Gerais, Brazil. Mycotaxon 104: 385-390.

Song B, Hosagoudar VB. 2003. A list of Lembosia species based on the literature. Guizhou Science

Ab 93-101-

MYCOTAXON

January-March 2017— Volume 132, pp. 241

http://dx.doi.org/10.5248/132.241

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT—Mycotaxon is pleased to announce another new species distribution paper to

our ‘web-list’ page covering clavarioid fungi in Brazil (by Meiras-Ottoni, Araujo-Neta &

Gibertoni). This brings to 124 the number of free access mycobiotas now available on the

Mycotaxon website: http://www.mycotaxon.com/resources/weblists.html

SOUTH AMERICA

Brazil

ANGELINA DE MEIRAS-OTTONI, LipIA SILVA ARAUJO-NETA & TATIANA

BAPTISTA GIBERTONI. A checklist of clavarioid fungi (Agaricomycetes)

recorded in Brazil. 22 p.

ABSTRACT—Based on an intensive search of literature about clavarioid fungi

(Agaricomycetes: Basidiomycota) in Brazil and revision of material deposited in

Herbaria PACA and URM, a list of 195 taxa was compiled. These are distributed

into six orders (Agaricales, Cantharellales, Gomphales, Hymenochaetales,

Polyporales and Russulales) and 12 families (Aphelariaceae, Auriscalpiaceae,

Clavariaceae, Clavulinaceae, Gomphaceae, Hymenochaetaceae, Lachnocladiaceae,

Lentariaceae, Lepidostromataceae, Physalacriaceae, Pterulaceae, and Typhulaceae).

Among the 22 Brazilian states with occurrence of clavarioid fungi, Rio Grande do

Sul, Parana and Amazonas have the higher number of species, but most of them

are represented by a single record, which reinforces the need of more inventories

and taxonomic studies about the group.