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VOLUME 105 JULY-SEPTEMBER 2008
Peter Roberts Heterochaete tenuicula sp. nov. ( p. 144, Fig 4.)
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Volume 105, pp. 1-5 July-September 2008
A new Pseudocercospora species on Passiflora setacea
ALEXEI C. DIANESE’, ANA M. COSTA’ & JOSE C. DIANESE?™
' alexei.dianese@cpac.embrapa.br 'Embrapa Cerrados, Br-020, Km 18, 73310-970 Planaltina, D.E, Brasil.
“Departamento de Fitopatologia, Universidade de Brasilia 70.910-900 Brasilia, D.F, Brasil.
Abstract — A new Pseudocercospora species, P. passiflorae-setaceae, was detected causing severe leaf spotting on Passiflora setacea in Brasilia, Distrito Federal, Brazil. The new species is described, and differences from other Pseudocercospora species on Passiflora are discussed.
Key words — cercosporoid fungi, Mycosphaerellaceae, tropical fruits, wild passion- fruit
Introduction
Native Passiflora L. species are common in the Cerrado as well as in other Brazilian Biomes (Cervi 1997, Nunes & Queiroz 2001, Vieira & Carneiro 2004). Among them P setacea DC. deserves attention being now domesticated for fruit production. Mendes et al. (1999) listed fungal species associated with Passiflora in Brazil, but among them species of Pseudocercospora Speg. were not mentioned. Crous & Braun (2003) revised the names of hyphomycetes previously referred to as Cercospora Fresen. and Passalora Fr. and published numerous new combinations with updated descriptions of known cercosporoid fungi including those on Passiflora species |viz., two Passalora species (Passalora biformis (Peck) U. Braun & Crous and P. fuscovirens (Sacc.) U. Braun & Crous), four Cercospora species (C. granadillae Chupp, and three belonging to C. apii Fresen. s. lat. (viz., C. passifloricola Chupp, C. regalis Tharp, C. truncatella G.E. Atk.), and three Pseudocercospora species (P. calospilea (Syd.) Deighton, P. passiflorae U. Braun & Crous, and P. stahlii (F. Stevens) Deighton)] (Chupp 1954, Deighton 1976, Ellis 1976, Crous & Braun 2003). However, the fungus on P. setacea is quite distinct from the Cercospora and Passalora species mentioned, which possess conidiogenous cells and conidia conspicuously
' Corresponding author: jcarmine@unb.br
2 ... Dianese, Costa & Dianese
cicatrized. Furthermore, the new species does not coincide with the known Pseudocercospora species occurring on hosts of the Passifloraceae (Chupp 1954, Deighton 1976, Braun et al. 1999, Crous & Braun 2003, Farr et al. 2008). The specimen studied revealed unique features sufficient to be described as a new Pseudocercospora species.
Materials and methods
Leaves of P. setacea showing strong yellow leaf spotting were collected from a passion-fruit nursery at Embrapa Cerrados in Brasilia. The sample was dried, numbered and deposited in the Mycological Collection of the Herbarium of the University of Brasilia (UB- Col. Micol.). Studies under the stereomicroscope were followed by observations of squash preparations and sections made with a freezing microtome. The morphological features were described, measured, and documented using a Leica DM 2500 microscope coupled with a Leica DFC 490 digital camera connected to a microcomputer. Image capture, editing, and measurements were made with the help of Leica QWin V3 software. In some cases, the samples were stained with lacto-glycerol cotton blue and the slides sealed with nail polish, but most of the photographic work was done without staining using Nomarski optics. A minimum of fifty replicates of spore and hyphal measurements were made.
Taxonomy
Pseudocercospora passiflorae-setaceae A.C. Dianese, A.M. Costa & Dianese, sp. nov. Figs. 1-11. MycoBank MB 511598
Pseudocercosporae passiflorae similis, sed stromatibus majoribus, (49-) 60-166 (-195) um diam., hyphis superficialibus cum conidiophoris solitariis evolutis et conidiis longioribus et latioribus, ad 129 x 5.5 um, ad 15-septatis.
Lesions adaxial, widespread, formed as irregularly scattered yellow solitary leaf spots, later coalescing into light brown necrotic areas with central black dots; grayish irregular downy spots on the abaxial surface. IMMERSED MYCELIUM septate, light brown, within the mesophyll, forming epiphyllous stromata and hypophyllous superficial mycelium. STROMATA (49-)60-(102)-165(-195) um diam, epiphyllous, subglobose to globose, brown to dark brown, subepidermal, erumpent, textura angularis. SUPERFICIAL MYCELIUM exclusively hypophyllous, strongly branched, intricate, abundant, light grayish brown to brown, giving rise to conidiophores and conidiogenous cells; hyphae 4-6 um diam, subhyaline to light bown, septate, smooth, thin-walled. EprpHyLLous CONIDIOPHORES 7-(14)-21(-31)x2.5-(4)-5 um, numerous, densely fasciculate,
Pseudocercospora passiflorae-setaceae sp. nov. (Brazil) ... 3
Figs. 1-4. Pseudocercospora passiflorae-setaceae on leaves of Passiflora setacea. 1. Yellow irregular leaf spots on the adaxial face. 2. Gray irregular leaf spots on the abaxial face. 3. Adaxial lesion showing small dots (stromata) in the central portion of the adaxial side. 4. Abaxial lesions covered by velvety gray superficial mycelium.
formed on stromata, geniculate, light brown, septate, smooth, HyPOPHYLLOUS CONIDIOPHORES Solitary, arising from superficial hyphae, lateral or terminal, aseptate, i.e. conidiophores reduced to conidiogenous cells. CONIDIOGENOUS CELLS integrated, terminal, 20-50 um long, sympodial; conidiogenous loci flattened, inconspicuous. Conip1a solitary, cylindrical, rarely obclavate- cylindrical, 43-(86)-129 x 3-(3.5)-5.5 um, 5-15-septate, solitary, sub-hyaline to light brown, thin-walled, smooth; apex subacute to subobtuse; base truncate to somewhat obconically truncate, 3-4.5 um wide; hila neither thickened nor
darkened.
SPECIMEN EXAMINED. BRAZIL. DISTRITO FEDERAL: Planaltina. CENTRO DE PESQUISA AGROPECUARIA DO CERRADO (CPAC), Km 18, BR-20 HiGHWay NorTH, on living leaves of Passiflora setacea, 12 Feb 2008, leg. Alexei de Campos Dianese 95, holotype. (UB- Col. Micol. 20872).
4 ... Dianese, Costa & Dianese
Figs. 5-6. Pseudocercospora passiflorae-setaceae on leaves of Passiflora setacea. Cross section of stromata with conidiophores on infected leaves showing the trans-mesophyllic mycelium (black arrows), and also the emerging hypophyllous superficial mycelium (white arrow).
Figs. 7-10. Pseudocercospora passiflorae-setaceae. 7. Hypophyllous intricate superficial mycelium. 8. Branching of the superficial mycelium with terminal conidiogenous cells. 9. Superficial mycelium (left arrow) bearing a geniculate conidiophore (right arrow) with a 10-septate conidium. 10. Conidia.
Discussion
All three known Pseudocercospora species on Passiflora are easily segregated from the new species, first of all, because they all lack an external hypophyllous mycelium with solitary conidiophores. Due to well-developed epiphyllous stromata with fasciculate conidiophores and similar conidia, Pseudocercospora passiflorae resembles P._ passiflorae-setaceae, but the stromata are much smaller, and the conidia are shorter and narrower, usually 30-80 x 2-4 um, only with up to 10 septa (Crous & Braun 2003). The second species on Passiflora, Pseudocercospora stahlii, was originally described from Puerto Rico as Helminthosporium stahlii F. Stevens in 1917, and later found in the Virgin Islands and Dominican Republic. This species is also known from 16 countries in Asia and Australia mainly affecting Passiflora foetida (Hsieh & Goh 1990, Hyde & Alcorn 1993, Liu 1977, Guo t al. 1998, Shaw 1984, Thaung 1984, Zhuang 2001). Pseudocercospora stahlii differs from P. passiflorae-setaceae by having amphigenous fasciculate conidiophores, smaller stromata (20-40 um), and short, broad conidia with up to seven septa (Deighton 1976). Finally,
Pseudocercospora passiflorae-setaceae sp. nov. (Brazil) ... 5
P. calospilea is clearly different from the new species by its smaller hypophyllous stromata (25-35 um diam.), hyaline to subhyaline, cylindrical-obclavate, 1-7- septate, smaller conidia (20-60 x 2-4 um). Based on the clear morphological differences, the introduction of a new species for the Pseudocercospora on Passiflora setacea is justified.
Acknowledgements
The authors thank Embrapa, CNPq and Fundacao Banco do Brasil for financial support, and Prof. Mariza Sanchez for assistance with the herbarium work, and C.A. Inacio for help with the photographic work. Also sincere thanks are given to Drs. Uwe Braun and Roland Kirschner for the pre-submission reviews of our manuscript.
References
Cervi AC. 1997. Passifloraceae do Brasil. Estudo do género Passiflora L., Subgénero Passiflora. Fontqueria 14:1-92.
Chupp C. 1954. Monograph of the fungus genus Cercospora. Published by the Author: Ithaca, New York (USA). 667 pp.
Crous PW, Braun U. 2003. Mycosphaerella and its anamorphs - 1. Names published in Cercospora and Passarola: Centraalbureau voor Schimmelcultures: Ultrecht (The Nederlands). 571 pp. Deighton FC. 1976. Studies on Cercospora and allied genera. VI. Pseudocercospora Speg., Pantospora
Cif. and Cercoseptoria Petr. Mycological Papers 140:1-168.
Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute: Kew, Surrey (England). 507 pp.
Farr DE, Rossman AY, Palm ME, McCray EB. 2008. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved February 14, 2008, from http://nt.ars-grin. gov/fungaldatabases/.
Guo Y-L, Liu X-J, Hsieh W-H. 1998. Pseudocercospora. Flora Fungorum Sinicorum. Vol. 9. Science Press: Beijing (China). 474 pp.
Hsieh W-H, Goh T-K. 1990. Cercospora and Similar Fungi from Taiwan. Maw Chang Book Co.: Taipei (Taiwan). 376 pp.
Hyde KD, Alcorn JL. 1993. Some disease-associated microorganisms on plants of Cape York Peninsula and Torres Strait Islands. Australasian Plant Pathology 22: 73-83.
Liu PSW. 1977. A supplement to a host list of plant diseases in Sabah, Malaysia. Phytopathological Papers 21: 1-49.
Mendes MAS, Silva VL, Dianese JC, Ferreira MASV, Santos CEN, Neto EG, Urben AEF, Castro C. 1998. Fungos em Plantas no Brasil. Embrapa - SPI / Embrapa - Cenargen: Brasilia (Brazil). 569 pp.
Nunes TS, Queiroz LP. 2001. A Familia Passifloraceae na Chapada Diamantina, Bahia, Brasil. Sitientibus Série Ciéncias Bioldgicas 1: 33-46.
Shaw DE. 1984. Microorganisms in Papua New Guinea. Department of Primary Industry Research. Bulletin 33: 1-344.
Thaung MM. 1984. Some fungi of Cercospora complex from Burma. Mycotaxon 19: 425-452.
Vieira MLC, Carneiro MC. 2004. Passiflora spp. Passionfruit. In: Litz, R. Biotechnology of fruit and nut crops. Oxford, CABI: Wallingford. p. 436-453.
Zhuang W-Y. 2001. Higher Fungi of Tropical China. Mycotaxon Ltd., Ithaca, NY. 485 pp.
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Volume 105, pp. 7-10 July-September 2008
Type studies on South American Strophariaceae: 2. Pholiota trinitensis is transferred to Stropharia
VAGNER G. CORTEZ
cortezvg@yahoo.com.br Universidade Federal do Rio Grande do Sul Programa de Pés-graduacdo em Botanica Av. Bento Goncalves, 9500, 91501-970, Porto Alegre, RS, BRAZIL
Abstract — The holotype of Pholiota trinitensis (= Hypholoma trinitensis) was examined, and the presence of numerous acanthocytes in its rhizomorphs necessitates its transfer to the genus Stropharia. The new combination, S. trinitensis, is proposed. The taxonomic discussion is accompanied by a brief description and illustrations of relevant microscopic features.
Key words — Agaricales, Naematoloma, neotropical mycobiota, nomenclature
This new contribution to type studies on South American Strophariaceae Singer & A.H. Sm. (Cortez 2008) presents results from the type study of Pholiota trinitensis. Although originally described from Central America (Trinidad Isl].), there are known records for the species from Brazil (Pegler 1997 from Sao Paulo State; de Meijer 2006 from Parana State; Bononi et al. 2007 from Mato Grosso do Sul State) and Argentina (Lechner et al. 2006). This species was described initially in Pholiota (Fr.) P. Kumm., later transferred to Hypholoma (Fr.) P. Kumm. (Pegler 1983), and then to Naematoloma P. Karst. (Singer 1986). However, the characters listed by Dennis and Pegler suggest that the species belongs to Stropharia (Fr.) Quél. following the current concept proposed by Cortez & Silveira (2008). The author therefore considered it necessary to examine the holotype to clarify its generic position.
The type specimens received from Kew were in fragments, consisting of pieces of two or three basidiomes. Abundant white rhizomorphs were easily seen on first observation. Pegler (1983), who studied both type and fresh specimens, provided a detailed macroscopic description that noted a 30-75 mm, convex to umbonate, cinnamon-brown to ochraceous-buff, hygrophanous pileus with white velar remnants appendiculate on the margin; adnate, pale grey to tobacco-brown, moderately crowded lamellae; a 30-50 x 5-8 mm, cylindrical, ochraceous cinnamon, striate stipe with abundant white rhizomorphs at the
8 ... Cortez
base; a cream colored, fibrillose to floccose veil forming an indistinct annular zone on upper stipe; and a dark maroon.
Prior to examination, thin sections of dried basidiomes from the holotype were mounted in 5% KOH and 1% Congo red solutions. The following characters were observed:
BASIDIOSPORES 6-7.5 X 3.7—4.6(—5) wm, Q= 1.45-1.78, Q = 1.65, n= 42, elliptic to phaseoliform in side view, ovoid to elliptic in frontal view, with a smooth and thick wall, and a reduced germ-pore (Fig. 4); BAsIDIA 17-21 x 6-7.5 um, subclavate, with a medial constriction, 4-spored (Fig. 3); PLEUROCYSTIDIA 38-59 x 10-14.2 wm, as chrysocystidia, lageniform to ventricose, with rounded apex, not mucronate, thin-walled, with greenish amorphous contents (Fig. 2); CHEILOCYSTIDIA not found, probably collapsed - according to Pegler (1983) they are 22-27 x 10-17 um, pyriform to globose, hyaline, and thin-walled, not of the chrysocystidia type; PILEIPELLIS composed of hyaline, smooth and thin-walled hyphae, 6-9.5 um diam. on a hypodermium formed by brownish, inflated hyphae, 14-25 wm diam. (Fig. 1); HYMENOPHORAL TRAMA subregular, composed by yellowish interwoven hyphae; CAULOCYSTIDIA 27-47 x 9-12 um, as chrysocystidia, scattered or in fascicles on stipe apex, and similar to the pleurocystidia; ACANTHOCYTES very abundant on rhizomorphs surface (Fig. 5):
The combination of the above characters makes generic assignment in Strophariaceae difficult, for the spore print suggests Pholiota while the subcellular hypodermium is more closely related to Hypholoma (= Naematoloma). However, the presence of acanthocytes easily supports placement of the species in Stropharia. This morphological feature, which is unique to the genus (Farr 1980), has been recently useful in diagnosing other Stropharia species (Cortez 2008; Cortez & Silveira 2007, 2008; Desjardin & Hemmes 2001; Norvell & Redhead 2000). Recent research (Luo et al. 2006) suggests that in S. rugosoannulata Farl. ex Murrill acanthocytes function as a nematode- attacking device unique to the genus.
Pholiota trinitensis must be compared with other neotropical members of Stropharia previously classified in Pholiota section Albivelatae A.H. Sm. & Hesler, including S. apiahyna (Speg.) Cortez & R.M. Silveira and S. varzeae (Singer) Cortez. Stropharia apiahyna differs in producing a smaller pileus (< 30 mm), a membranous annulus, and longer cheilocystidia (Cortez & Silveira 2008). Stropharia varzeae, which also has membranous annulus, produces cheilocystidia as chrysocystidia (Cortez 2008). Stropharia acanthocystis Cortez & R.M. Silveira, from Araucaria angustifolia (Bertol.) Kuntze forests in south Brazil, is similar in basidiospore size and cheilocystidial shape. However, in S. acanthocystis the cheilocystidia have thickened walls, pleurocystidia are
Stropharia trinitensis comb. nov. (Brazil) ... 9
Figs. 1-5. Stropharia trinitensis - Holotype: K(M) 147682. 1. Pileipellis and hypodermium. 2. Pleurocystidia. 3. Basidia. 4. Basidiospores 5. Acanthocytes.
clavate, caulocystidia are of three types (leptocystidia, chrysocystidia, and acanthocytes), and (unique within Stropharia) acanthocytes are also present as hymenial cystidia (Cortez & Silveira 2007).
Naematoloma amazonicum Singer, described by Singer (1973) from Brazil, was considered by Pegler (1983, as Hypholoma) and Singer (1986, as Naematoloma) as a synonym of P. trinitensis. This synonymy only will be confirmed after the study of the holotype of N. amazonicum.
Based on all above cited characters, P. trinitensis is considered a good species of Stropharia and the new combination is introduced:
10 ... Cortez
Stropharia trinitensis (Dennis) Cortez, comb. nov. MycoBank MB 511476 BasionyM: Pholiota trinitensis Dennis, Kew Bull. Add. Ser. 3: 467, 1970. SynonyMs: Hypholoma trinitense (Dennis) Pegler, Kew Bull. Add. Ser. 9: 509, 1983. Naematoloma trinitense (Dennis) Singer, Agar. Mod. Taxon.: 564, 1986.
EXAMINED SPECIMEN: TRINIDAD ISL. St. Augustine, 27.IX.1949, R.W.G. Dennis 78 —- K (M) 147682 (holotype).
Acknowledgements
The author is grateful to the herbarium Kew for loan of the type collection. Special thanks to Dr. Maria A. Neves (Universidade Estadual de Feira de Santana, Brazil) and Dr. Lorelei L. Norvell (Pacific Northwest Mycology Service, USA) for critical review of the manuscript. CNPq (Brazil) is acknowledged for financial support.
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Cortez VG. 2008. Type studies on South American Strophariaceae. 1. Pholiota varzeae, from Brazilian Amazon. Mycotaxon 103: 137-140.
Cortez VG, Silveira RMB. 2007. A new species of Stropharia with hymenial acanthocytes. Mycologia 99: 13 5= 158.
Cortez VG, Silveira RMB. 2008. The agaric genus Stropharia (Strophariaceae) from Rio Grande do Sul State, Brazil. Fungal Diversity 32: 31-57.
de Meijer AAR. 2006. Preliminary list of the macromycetes from the Brazilian State of Parana. Bol. Mus. Bot. Mun., Curitiba 68: 1-55.
Dennis RWG. 1970. Fungus Flora of Venezuela and adjacent countries. Kew Bull. Add. Ser. 3: 125318
Desjardin DE, Hemmes DE. 2001. Agaricales of the Hawaiian Islands - 7. Notes on Volvariella, Mycena Sect. Radiatae, Physalacria, Porpoloma and Stropharia. Harv. Pap. Bot. 6: 85-103.
Farr DE 1980. The acanthocyte, a unique cell type in Stropharia (Agaricales). Mycotaxon 11: 241-249.
Lechner BE, Wright JE, Popoff O. 2006. New taxa and new records for Argentina of fungi from Iguazu National Park, Misiones. Fungal Div. 21: 131-139.
Luo H, Li X, Li G, Pan Y, Zhang K. 2006. Acanthocytes of Stropharia rugosoannulata function as a nematode-attacking device. Appl. Environ. Microbiol. 72: 2982-2987.
Maerz A, Paul MR. 1930. A Dictionary of Color. New York (USA): McGraw-Hill, 207pp.
Norvell LL, Redhead SA. 2000. Stropharia albivelata and its basionym Pholiota albivelata. Mycotaxon 76: 315-320.
Pegler DN. 1983. Agaric Flora of the Lesser Antilles. Kew Bull. Add. Ser. 9: 1-668.
Pegler DN. 1997. The Agarics of Sao Paulo, Brazil. London (UK): HMSO/Royal Botanic Gardens, Kew, 68pp.
Singer R. 1973. Diagnoses Fungorum Novorum Agaricalium HI. Beih. Sydowia 7: 1-106.
Singer R. 1986. The Agaricales in Modern Taxonomy. 4"° ed. Koenigstein (Germany): Koeltz, 981pp.
MYCOTAXON
Volume 105, pp. 11-18 July-September 2008
Ambispora brasiliensis, anew ornamented species in the arbuscular mycorrhiza-forming Glomeromycetes
BRUNO TOMIO GOTO!, LEONOR CosTA MAIA! & FRITZ OEHL?
™brunogoto@hotmail.com ‘Departamento de Micologia, CCB, Universidade Federal de Pernambuco Av. Prof. Nelson Chaves s/n, Cidade Universitaria, 50670-420, Recife, PE, Brazil
*Zurich-Basel Plant Science Center, Institute of Botany, University of Basel Hebelstrasse 1, 4056 Basel, Switzerland
Abstract — A new dimorphic fungal species in the arbuscular mycorrhiza-forming Glomeromycetes was found in the Cerrado biome in Minas Gerais (Southeast Brazil) and is here described from acaulosporoid glomerospores under the epithet Ambispora brasiliensis. Glomoid glomerospores of this species were found only once, connected to a germinating acaulosporoid glomerospore. The acaulosporoid glomerospores are formed on a pedicel branching from the neck of a sporiferous saccule. They are hyaline to pale yellow in color, globose, 62-95 um in diameter, to subglobose to oval, 59-88 x 69-100(-118) um, and with three spore walls. Crowded irregular pustules, 2.4-7.0(-10.0) um high and 4.2—9.8(—15.0) um wide, are formed on the surface of the outer wall. The glomoid glomerospores are hyaline to subhyaline, globose, 25-30 um in diameter, and have a bi-layered spore wall without ornamentation. The acaulosporoid glomerospores of the new fungus can easily be distinguished from all other species in the genus Ambispora by spore size and by the distinct pustulate ornamentation.
Key words — Glomeromycota, Ambisporaceae, bimorphic, germination
Introduction
Species of the genus Ambispora C. Walker et al. of the arbuscular mycorrhiza (AM) forming Ambisporaceae C. Walker et al. (Glomeromycetes) can form acaulosporoid and glomoid glomerospores (Spain et al. 2006, Walker et al. 2007a, Walker 2008). For some species only the acaulosporoid morph is known (e.g. A. jimgerdemannii (N.C. Schenck & T.H. Nicolson) C. Walker), for others only the glomoid morph (e.g. A. leptoticha (N.C. Schenck & G.S. Sm.) C. Walker et al. and A. callosa (Sieverd.) C. Walker et al.), while at least A. appendicula (Spain et al.) C. Walker, A. gerdemannii (S.L. Rose et al.) C. Walker et al. and A. fennica C. Walker et al. definitely form both morphs (Spain et al. 2006, Walker et al. 2007a, Walker 2008). There has been some confusion regarding the valid taxonomic nomenclature of Ambisporaceae and its type genus Ambispora in the
12 ... Goto, Maia & Oehl
recent literature resulting from past incorrect re-combinations and synonymies (see review by Spain et al. 2006), or obviously illegitimate names attributed by Spain et al. (2006) at the genus level (see Walker et al. 2007a,b, 2008).
Acaulosporoid glomerospores of Ambispora can quite easily be recognized by their morphology (Spain et al. 2006) whereas the glomoid morph might be mistaken with glomerospores of the Glomeraceae. Indeed, Walker et al. (2007a) identified Glomus callosum Sieverd. as belonging to Ambispora only using molecular biological techniques.
Acaulosporoid glomerospores of an undescribed Ambispora species were detected in rhizospheric soil from a natural ecosystem of the Brazilian Cerrado biome in Serra do Cipé (Minas Gerais State, Southeast Brazil). The species, characterized by a distinct pustulate ornamentation on the outer spore wall, is presented here under the epithet Ambispora brasiliensis.
Material and methods
Soil sampling and culturing of AM fungi
Soils were sampled in the Cerrado biome of Serra do Cip6o (Minas Gerais State, Brazil) from the rhizosphere of typical native plant vegetation, mainly consisting of Vellozia caruncularis Mart. ex Seub., in July 2004. The site is located at 19° 06'S and 43° 35°W. Soil pH (H,O) was 4.7.
For culturing the native AM fungal communities, 50 g of the field soil were mixed with 400 g autoclaved sand-vermiculite substrate (1:1 w/w, pH 5.5) and filled into 500 ml pots. Sorghum bicolor (L.) Ménch was used as bait plant. The culture pots were maintained at the greenhouse of the Department of Mycology, Universidade Federal de Pernambuco, Recife. Additionally, multiple glomerospores of the new species were separated and used as infective propagules in single species cultures using the same pot size and sand-vermiculite culture substrate as described above. So far, all trials failed to establish a successful symbiosis with the new AMF species both in bait cultures and in single species cultures.
Morphological analyses
Acaulosporoid glomerospores were extracted from field soils and the culture substrates by wet sieving (Gerdemann & Nicolson 1963) and sucrose centrifugation (Jenkins 1964). The spores were thereafter isolated under a stereo microscope and mounted in polyvinyl-lacto-glycerol (PVLG), PVLG + Melzer’s reagent (Brundrett et al. 1994) and in water, respectively. For the description of the glomerospores and the spore walls, the terminology of Spain et al. (2006) recently presented for species of the genus was used. We use the term glomerospore for all spores formed by species of the Glomeromycetes, as proposed by Goto & Maia (2006).
Ambispora brasiliensis sp. nov. (Brazil) ... 13 Description of the new species
Ambispora brasiliensis B.T. Goto, L.C. Maia & Oehl, sp. nov. Figs. 1-9 MycoBank MB 511612
Sporocarpia ignota. Sporae dimorphae; sporae acaulosporoideae hyalinae vel subhyalinae, globosae (62-95 um in diametro) vel subglobosae vel ovales (59-88 x 69-100(-118)) um, cum ternibus tunicis stratis pluribus, in solo singulatim formatae appendice lateraliter ex hypha terminata in sacculo sporangifero; sporae cum tunica exterior, tunica media et tunica interior: tunica exterior tunica mediaque coniuncta sacculi et hyphae; tunica exterior stratis tribus in totum 2.7-3.8 um crassa, stratum exterius tunicae exterioris hyalinum, evanescens, 0.4-0.7 um crassum; stratum secundum hyalinum at flavo-pallidum, 2.2-3.2 um crassum, pustulibus irregularibus ornatum altis 2.4—7.0(-10)um et 4.2-9.8(-15) um latis ornatum; stratum interius tunicae exterioris hyalinum at flavor-pallidum, subtile; tunica media duobus stratis hyalinis in totum 2.8-5.2 um crassa; tunica interior de novo formans stratis duabus vel tribus, in totum 2.2-2.9 um crassa. Sporae glomoidae hyalinae vel subhyalinae, 25-30 um in diameterum, stratis duobus. Holotypus hic depositus 78879: URM.
EryMo.ocy: brasiliensis referring to Brazil, the country where the species was found.
GLOMEROSPORE FORMATION. Sporocarp formation is unknown. The species is dimorphic producing acaulosporoid and glomoid morphs. Acaulosporoid glomerospores form singly in soils upon a short pedicel arising laterally on the tapering hyphal neck of a sporiferous saccule (Figs. 1-3). The acaulosporoid glomerospores form three spore walls: an outer wall (ow), a middle wall (mw) and an inner wall (1w) (Figs. 4-9). Glomoid glomerospore formation was observed once on a hypha connected to a germinating acaulosporoid morph.
SPORIFEROUS SACCULE OF ACAULOSPOROID GLOMEROSPORESare hyaline to light yellow, globose to subglobose, 65-115 um in diameter and formed terminally on a hypha (Figs. 1-2). The saccule wall generally is bi-layered with a hyaline to light yellow, rapidly degrading, evanescent outer layer (0.5-0.9 ttm thick) and a hyaline to subhyaline, semi-persistent inner layer (0.6-1.5 um thick). The hyphal neck of the sporiferous saccule is 10-17 um wide at the saccule base tapering to 8-12 um at the point where the pedicel of the acaulosporoid glomerospore branches, narrowing further to 5-8 um at a distance of 40-120 um from the pedicel. A septum arises in the tapering hyphal neck distal to the saccule where the pedicel forms. The sporiferous saccule generally collapses and detaches from the mature spore while the pedicel often persists on the glomerospore.
ACAULOSPOROID GLOMEROSPORES form singly on a short hyphal pedicel (Fig. 3). The spores are globose (62-95 um in diameter) to subglobose to oval, 59-88 x 69-100(-118), hyaline to subhyaline in water and generally become light yellow to ochre-yellow in PVLG-based mountants. The two wall layers of the saccule continue on the glomerospore differentiating into two separate glomerospore walls: a two to three layered outer wall (ow) and a bi-layered middle wall (mw).
14 ... Goto, Maia & Oehl
A two-to-three layered inner wall (tw) forms de novo, presumably after the spore pore has been closed by the inner layer of the middle wall (mw12) which differentiates from MwL1. The rw functions as a germinal wall.
OUTER WALL generally consists of three layers (Figs. 4-8). The outer layer (OWL1) is hyaline to subhyaline, evanescent, 0.4-0.7 um thick, and often is difficult to detect, as it tightly adheres to owL2 (Fig. 8). OWL2 is hyaline to light yellow and in total 2.2-3.2 um thick and crowded with conspicuous, convex pustules (Fig. 3-8). The pustules vary greatly in shape and size and are generally 2.4—7.0(-10) um wide and 4.2-9.8(-15) high (Figs. 4-7). The inner layer (OWL3) is thin (0.3-0.6 um), hyaline to subhyaline and tightly attached to owL2 making it difficult to detect. In older spores, the outer wall might degrade completely, but usually some fragments remain on the surface of the middle wall.
MIDDLE WALL is bi-layered and in total 2.8-5.2 um thick (Figs. 4-6; Figs. 8-9). MWLI is hyaline, 1.3-2.6 um thick and continuous with the persistent inner layer of the pedicel. A septum arising in the pedicel from MWwL1 during spore formation was rarely detected. MWL2 is also hyaline, 1.5-2.6 tim thick, tightly adherent to MwL1 and generally closes the pore of the pedicel.
INNER WALL is hyaline and three-layered but generally only the central layer (IWL2; Figs. 4-6; Fig. 8—9) or the inner two layers (IwL2 and IWL3) are visible. The outer layer IWL1 is 0.3-0.6 um and difficult to detect, as it is tightly adherent to the central layer IwL2 which is 1.5-2.2 um thick. The innermost layer is 0.4—0.8 tum thick and also tightly adherent to rwL2 but sometimes separates slightly showing several thin folds when pressure is applied.
PEDICEL OF ACAULOSPOROID GLOMEROSPORE branches from the hyphal neck of the sporiferous saccule in about 40-130 um distance from the globose saccule terminus. The pedicel is (5—)7—16(-25) um long, and 8-13 um wide at the glomerospore base tapering to 5-9(-13) um at the connection to the hypha. The two wall layers of the hyphal neck of the sporiferous saccule differentiate into the outer and the middle wall of the pedicel (ow and Mw). On mature spores, the pedicel often persists resembling a subtending hypha of glomoid glomerospores (Fig. 5). Sometimes only the ow (Fig. 5) or the Mw (Fig. 6) of the pedicel persist on the spore; rarely both pedicel walls are found on mature glomerospores. The ow sometimes forms a collar (Fig. 7) on the glomerospore (8.5-14 um in diameter). The ow, if present on the pedicel, is 1.2-2.5 um thick at the glomerospore base tapering to 0.5-0.9 um towards the saccule neck. The Mw is 1.3-2.1 um at the glomerospore base tapering to 0.9-1.5 um.
GLOMOID GLOMEROSPORES were detected only once on a connecting hypha about 250 um from a germinating acaulosporoid glomerospore. ‘The hyaline to subhyaline glomoid glomerospores were bi-layered and about 25-30 um in diameter. The glomerospore wall was 1.9-2.5 um thick in total, and the
Ambispora brasiliensis sp. nov. (Brazil) ... 15
Dee ser
Figs. 1-9: Ambispora brasiliensis forming acaulosporoid glomerospores with three walls (outer wall ow, middle wall mw and inner wall rw) upon a short pedicel arising laterally on the neck of a sporiferous saccule. Fig. 1. Acaulosporoid glomerospore and sporiferous saccule (sac) attached. Fig. 2. Saccule with two layers: evanescent outer layer continuous with ow of the spore and semi- persistent inner layer continuous with MwL1 of the acaulosporoid spore. Fig. 3. Pedicel (pcL) and degrading saccule. Figs. 4-6. Irregular pustulate ornamentation (ORN) on middle layer of ow; MW has two layers (MwLI + Mw12). Fig. 5. Pcl wall continuous with ow. Fig. 6. Pcl wall continuous with MwLl. Fig. 7. Glomerospore pore and collar formed on ow. Figs. 8-9. Spore wall structure: ow with three layers (evanescent owLl, pustulate owL2; owL3 hardly detectable), mw bi-layered (MwLI1 + MWL2); only central layer evident on Iw.
16 ... Goto, Maia & Oehl
subtending hypha was 2.0-3.1 «sm wide at the base of the glomerospore, where a septum was not visible.
GERMINATION was observed once on an acaulosporoid glomerospore, where it initiated on the rw below the pedicel collar. Three germ tubes (2.4-4.0 um wide) emerged from the center of a lobed germination structure, which was 34 x 15 um in diameter and consisted of 6-7 lobes that were each 12-15 um long and 5-8 um wide. The germ tubes had grown through the pedicel collar and branched in about 200-250 ttm distance from the glomerospore. However, it was not clearly discernable if the lobed germination structure was formed between Iw and Mw or between the mw and the collar formed by the pedicel on the ow.
MYCORRHIZAL FORMATION — unknown.
DisTRIBUTION — Brazil: so far only found in the Cerrado biome of Serra do Cipo, Minas Gerais State. SPECIMENS EXAMINED — BRAZIL. Minas Gerais. Serra do Cipé. HOLOTYPE (54- 5401) and type specimens (54-5402) deposited at URM, Recife, Pernambuco, Brazil (accession numbers URM 78879 and 78880). Isotype (54-5402) specimens deposited at OSC, Corvallis, Oregon, USA (accession number OSC# 134501); and 54-5406 & 54- 5407, deposited at Z+ZT, Zurich, Switzerland (accession number ZT MYC 159).
Discussion
The new fungal species, Ambispora brasiliensis, can easily be identified by its acaulosporoid morph and can be distinguished from all other Ambispora species forming acaulosporoid morphs by glomerospore size and by the distinct pustular ornamentation on the acaulosporoid glomerospore walls.
Acaulosporoid glomerospores of other genera in the Glomeromycetes — namely Acaulospora, Archaeospora and Otospora (Gerdemann & Trappe 1974, Morton & Redecker 2001, Palenzuela et al. 2008) — cannot be confused with A. brasiliensis. Spores of Archaeospora and Otospora have two walls (Palenzuela et al. 2008) contrasting with Acaulospora and Ambispora, which have three spore walls (Spain et al. 2006, Walker et al. 2007a, Palenzuela et al. 2007). Glomerospores of Acaulospora are not formed on a pedicel (= appendix) as known for the acaulosporoid glomerospore formation of Ambispora. Furthermore, in Acaulospora the middle spore wall forms de novo (e.g. Oehl et al. 2006) and is not continuous with the wall of the sporiferous saccule as noted for Ambispora brasiliensis and other Ambispora species (Spain et al. 2006, Walker et al. 2007a). Finally, only three Acaulospora species are known to form projections on the - outer spore wall: Ac. spinosa, Ac. tuberculata and Ac. denticulata, which form — according to their names — either spiny, tuberculate or dentate ornamentations (Walker & Trappe 1981, Janos & Trappe 1982, Sieverding & Toro 1987) but not convex, irregular pustules as in A. brasiliensis.
Ambispora brasiliensis sp. nov. (Brazil) ... 17
We found a glomoid morph of this new species connected via hypha to a germinating acaulosporoid morph of A. brasiliensis only in one occasion. Such glomoid glomerospore formation during germination of acaulosporoid spores was also found for A. appendicula (Spain pers.comm. 2008; Goto & al. unpub. obs.). In addition to the glomoid morph, swollen hyphal tips on mycelia were also noted for A. appendicula (Schenck et al. 1984).
We were not able to culture A. brasiliensis but hope to conduct genetic analyses as soon as clean cultures are available. However, in the absence of genetic data, A. brasiliensis can be identified morphologically by observing its acaulosporoid glomerospores. As stated in the introduction, the glomoid morph of some of the Ambispora spp. is still unknown, and, in any case, Ambispora is more easily diagnosed by observing the acaulosporoid rather than the glomoid morph.
Acknowledgements
The authors acknowledge, in special, Dr. Ewald Sieverding (Institute of Plant Production and Agroecology in the Tropics, University of Hohenheim, Stuttgart, Germany) and Joyce L. Spain (Corvallis, Oregon USA), for reviewing the manuscript and making helpful comments and suggestions, and Dr. Nanuza Luiza de Menezes (Universidade de Sao Paulo, Brasil) for providing the opportunity to visit the Serra do Cipé, MG. This work was supported by the Conselho Nacional de Desenvolvimento Cientifico e Tecnoldgico (CNPq) and Coordenacao de Aperfeigoamento de Pessoal de Nivel Superior (CAPES) that provided, respectively, a research grant to Leonor C. Maia and a PhD scholarship to Bruno T. Goto. This work was also supported by the Universidade Federal de Pernambuco, which provided a grant to F. Oehl as ‘visiting professor’
Literature cited
Brundrett M, Melville L, Peterson L. 1994. Practical Methods in Mycorrhizal Research. Mycologue Publications, University of Guelph, Guelph, Ontario, Canada.
Gerdemann JW, Nicolson TH. 1963. Spores of mycorrhizal Endogone species extracted from soil by wet sieving and decanting. Trans. Br. Mycol. Soc. 46: 235-244.
Gerdemann JW, Trappe JM. 1974. The Endogonaceae in the Pacific Northwest. Mycologia Mem. No. 5. 76 p.
Goto BT, Maia LC. 2006 Glomerospores: a new denomination for the spores of Glomeromycota, a group molecularly distinct from Zygomycota. Mycotaxon 96: 129-132.
Janos DP, Trappe JM. 1982. Two new Acaulospora species from Tropical America. Mycotaxon 15: DES-522.
Jenkins WR. 1964. A rapid centrifugal-flotation technique for separating nematodes from soil. Plant Dis. Rep. 48: 692.
Morton JB, Redecker D. 2001. Two new families of Glomales, Archaeosporaceae and Paraglomaceae, with two new genera Archaeospora and Paraglomus, based on concordant molecular and morphological characters. Mycologia 93: 181-195.
Nicolson TH, Schenck NC. 1979. Endogonaceous mycorrhizal endophytes in Florida. Mycologia 71: 178-198.
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Oehl F, Sykorova Z, Redecker D, Wiemken A, Sieverding E. 2006. Acaulospora alpina, a new arbuscular mycorrhizal fungal species characteristic for high mountainous and alpine grasslands of the Swiss Alps. Mycologia 98: 286-294.
Palenzuela J, Ferrol N, Boller T, Azcén-Aquilar C, Oehl F. 2008. Otospora bareai, a new fungal species in the Glomeromycetes from a dolomitic shrub-land in the National Park of Sierra de Baza (Granada, Spain). Mycologia 100: 282-291.
Schenck NC, Spain J, Sieverding E, Howeler RH. 1984. Several new and unreported vesicular- arbuscular mycorrhizal fungi (Endogonaceae) from Colombia. Mycologia 76:685-699.
Sieverding E, Toro S. 1987. Acaulospora denticulata sp. nov. and Acaulospora rehmii sp. nov. (Endogonaceae) with ornamented spore walls. Angew. Bot. 61: 217-223.
Spain JL, Sieverding E, Oehl F. 2006. Appendicispora, a new genus in the arbuscular mycorrhizal- forming Glomeromycetes, with a discussion of the genus Archaeospora. Mycotaxon 97: 163-182.
Walker C. 2008. Ambispora and Ambisporaceae resurrected. Mycol. Res. 112: 297-298.
Walker C, Trappe JM. 1981. Acaulospora spinosa sp. nov. with a key of Acaulospora. Mycotaxon | Ped be eA
Walker C, Vestberg M, Demircik F, Stockinger H, Saito M, Sawaki H, Nishmura I, Schiissler A. 2007a. Molecular phylogeny and new taxa in the Archaeosporales (Glomeromycota): Ambispora fennica gen. sp. nov., Ambisporaceae fam. nov., and emendation of Archaeospora and Archaeosporaceae. Mycological Research 111: 137-153.
Walker C, Vestberg M, Schiissler A. 2007b. Nomenclatural clarifications in Glomeromycota. Mycol. Reés91 12532255)
MYCOTAXON
Volume 105, pp. 19-22 July-September 2008 Lichens of Malatya, Elazig and Adiyaman provinces (Turkey)
MEHMET CANDAN* & AYSEN OZDEMIR TURK
mecandan@anadolu.edu.tr e& aturk@anadolu.edu.tr Department of Biology, Faculty of Science, Anadolu University Eskisehir, Turkey
Abstract—The lichenized and lichenicolus fungi of Malatya, Elazig and Adiyaman provinces of Turkey, based on the authors’ collections and literature records, are listed, of which 20 are new for Turkey and 187 are new for the study area. As a consequence of this study, there are 315 infrageneric taxa recorded from these provinces. The full Checklist can be viewed at http://www.mycotaxon.com/resources/weblist.html
Key words—lichenized fungi, new records, biodiversity
An increasing number of studies on the lichen flora of Turkey have been carried out in the last two decades, most focused on central, western and northern Turkey where even today some areas remain unexplored. The present floristic study of the previously overlooked Malatya, Elazig and Adityaman provinces redresses to some extent this situation.
The first lichen records of six independent researchers from the study area, concerned with the distribution and uses of manna lichens undertaken during 1864-1897, are reviewed by Donkin (1981). Steiner (1921) reported 74 taxa from Elazig and Adiyaman provinces, based on Handell-Mazettis collections from Mesopotamia, Syria and Iraq. Szatala (1960) recorded 8 lichens from Elazig Province, as well as records from other parts of Turkey. Szatala’s paper (1960) based on the lichen collection of zoologist Victor Pietschmann, all contained lichen records from Syria and Iraq. Recently, Candan & Ozdemir Tiirk (2000) recorded 50 infrageneric taxa from eastern part of Malatya City. Other records for the study area are two exsiccates (Lumbsch & Feige 1999, John 1999), two by Degelius (1954), one by Steiner & Poelt (1982), one by Timdal (1991) and new lichenicolous fungi by Halici et al. (2007a,b) and Halici & Candan (2007).
As in other parts of Turkey, the natural vegetation of Malatya, Elazig and Adiyaman provinces has been subjected to anthropogenic stress for many years, mainly due to heavy grazing, afforestation, farming and urbanisation. Furthermore, many large dams have been built in southeastern Anatolia during the last 30 years and others are currently under construction. Hence, a relatively large portion of the study area is covered by dam lakes. As well as determining the current lichen
20 ... Candan & Tiirk
Figure 1. Map of the study area
diversity of Malatya, Elazig and Adiyaman provinces, this article also includes previous records from the region dating back to the 1860s. It is also hoped that this study will provide a database for monitoring the effects of the climate change on the lichen flora due the development of dams and their resultant lakes.
Stupy AREA: Malatya, Elazig and Adiyaman provinces are located in Anatolia and are part of the Irano-Turanian Phytogeographical Region of Turkey. According to Davis (1965), Malatya and Elazig are in the Upper Euphrates and Adiyaman is in the Mesopotamian area. The study area (Fig. 1) is located between 37°25’ 39°11’ N and 37°25’ 40°21’ E at altitudes ranging from 500 m in the south of Adryaman Province to 2650 m in the Malatya Mountains lying on the border of Adtyaman and Malatya provinces. The northern part of the study area, including Malatya and Elazig provinces, is mainly covered by highlands, the lowest altitudes of c. 800 m being found in valleys.
The study area also covers the eastern side of the Nurhak Mountains, the Malatya Mountains, the Maden Mountains and Akdag located in the Southeast Taurus Mountains, the highest altitudes being the summits of Akdag (2650 m), Bey (2545 m), Hazar (2347 m), Akcababa (2052 m) and Hasan (1864 m). The south side of Yama Mountain (2402 m), situated between Malatya and Sivas provinces, is also included in the study area. The Upper Firat, Tohma Stream, Siirgii Stream, Sultan Creek, Murat, and Dicle Valleys are located between the high mountains. Although Hazar Lake is the largest natural lake in the area, the lakes of Keban, Karakaya, Atatiirk, Siirgii and Cat Dams cover a much larger area collectively.
The rich topographical diversity of the study area provides varied climatic conditions, which are further modified the effects of the dam lakes; to date, no
Lichens of Malatya, Elazig, & Adiyaman... 21
research has been carried out to determine these effects. The study area has a Meditteranean climate. According to Emberger’s principles, Akman (1990) reported that precipitation-temperature coefficients (Q,) are 36.2, 39.6 and 75.4 and aridity indices (S) are 0.7, 0.5 and 0.2 for Malatya, Elazig and Adryaman provinces, respectively.
The dominant vegetation of the region is steppe in character. For anthropological reasons, no large forests are to be found in the study area; Quercus spp. are sparsely distributed. The most eastern distribution of Pinus brutia Ten. in Turkey is located at the junction of the borders of Kahramanmaras, Malatya and Adiyaman provinces which coincides with the southeast part of the study area. In spite of poor forest cover, there is intensive apricot (Prunus armeniaca L.) agriculture in some parts of Malatya Province and pistacio (Pistacia vera L.) agriculture in Adtyaman Province. Wild Prunus, Crataegus and Rosa spp. are also common in the study area, and Salix spp., Populus spp., and Platanus orientalis L. are found in the wet habitats such as valleys.
MeTHoboLoGy: This study is based on an evaluation of the authors’ lichen collections made between 2003 and 2005 and on literature sources relating to Malatya, Elazig and Adiyaman provinces of Turkey. The specimens were examined with a Leica MZ6 stereomicroscope and an Olympus BX51 microscope with standard identification methods for lichens and lichenicolous fungi (Purvis et al. 1992, Wirth 1995). Also TLC analyses of the specimen were carried out when needed. Voucher specimens are stored in ANES (Herbarium of Anadolu University, Anadolu University, Faculty of Science, Eskisehir, Turkey).
CONCLUSIONS: Twenty ofthe lichens in the Checklist are new for Turkey: Acarospora macrospora subsp. murorum, A. peliscypha, Aspicilia emiliae, Buellia imshaugii, B. pulverulenta, Caloplaca erodens, Dermatocarpon rivulorum, Fulgensia pruinosa, Immersaria usbekica, Lecanora laatokkaensis, L. reuteri, Placidiopsis custnani, Placidium tenellum, Polyblastia albida, Rinodina interjecta, R. luridata, R. roscida, Staurothele bacilligera, Thelidium incavatum, and Xanthoparmelia glabrans.
According to the literature, 128 lichens and lichenicolous fungi have been previously recorded from the study area. As a consequence of this study, this number has increased to 315. However, some early papers (Donkin 1981, Steiner 1921, Szatala 1960) provide only vague locality details (many of which have changed their names), making it difficult to evaluate distributional changes of lichens.
The study area is a transition region between the Mediterannean and the Irano-Turanian Phytogeographical Regions, the western and southwestern parts bordered by the former in Turkey where the distribution of Pinus brutia ends. This is why some authors delimit this region by the distribution of P. brutia while others delimit it by the Mediterranean sclerophyll forests, but both have very similar phytogeographical boundary.
22 ... Candan & Tiirk
Because of the interesting geographical position of the study area, lichens commonly distributed in Central Asia, Middle East and Europe also occur with local lichens, leading to a rich lichen diversity. Furthermore the altitude of the study area starts from 500 m in Adiyaman Province and 800 m in Elazig Province, and reaches higher than 2000 m in some localities. Therefore, high mountain lichens also make a significant contribution to the lichen flora of the area. However, lichen diversity is modified by the limited forest cover which reduces humid and shade loving species; most lichen epiphytes grow on free-standing trees and are nitrophilic and heliophilic.
Acknowledgements
We are grateful to Prof. Mark R. D. Seaward for linguistic revision, helpful comments and reviewing the manuscript, to Dr. P. Divakar for reviewing the manuscript and to the authors whose names are indicated in the List of Taxa. This study was funded by Anadolu University Scientific Research Project Unit (grant no. 031043).
Literature cited
Akman Y. 1990. [klim ve Biyoiklim. Ankara, Palme.
Davis P. 1965. Flora of Turkey. Vol: 1. Edinburgh, Edinburgh University Press.
Candan M, Ozdemir Tiirk A. 2000. Orduzu-Malatya bélgesi likenleri. OT Sistematik Botanik Dergisi 7: 219-230.
Degelius G. 1954. The lichen genus Collema in Europe. Symb. Bot. Upsal. 13 (2): 183, 329.
Donkin RA. 1981. Reports and Comments, The Manna Lichen: Lecanora esculenta. Anthropos 76: 562-572.
Halici1 MG, Candan M. 2007. Notes on some lichenicolous fungi from Turkey. Turkish Journal of Botany 31: 353-356.
Halici MG, Ozdemir Tiirk A, Candan M. 2007a. New records of pyrenocarpous lichenicolous fungi from Turkey. Mycotaxon 99: 201-206.
Halici MG, Atienza V, Hawksworth DL. 2007b. Two new Polycoccum (Dothideales, Dacampiaceae) species from Turkey. Mycotaxon 101: 157-163.
John V. 1999. Lichenes Anatolici Exsiccati. Arnoldia Fasc. 1-3 (No. 1-75): 44.
Lumbsch HT, Feige GB. 1999. Exsiccat. Lecanoroid lichens. Univ. Essen, Essen, Fasc. 5: 1-9.
Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM. 1992. The Lichen Flora of Great Britain and Ireland. London, Natural History Museum Publications in Association with The British Lichen Society.
Steiner J. 1921. Lichenes aus Mesopotamien un Kurdistan sowie Syrien und Prinkipo. Gesammelt von Dr. Heinrich Frh. v. Handell-Mazzetti (wissenschaftliche Ergebnisse der Expedition nach Mesopotamien 1910). Ann. des Naturhist. Museums in Wien 34: 1-68.
Steiner M, Poelt J. 1982. Caloplaca sect. Xanthoriella, sect. nov: Untersuchungen tiber Xanthoria lobulata-Gruppe (Lichenes, Teloschistaceae). Plant Systematics and Evolution 140: 151-177.
Szatala O. 1960. Lichenes Turcicae Asiaticae ab Victor Pietscmann collect. Sydowia 14: 312-325.
Timdal E. 1991. 41 monograph of the genus Toninia (Lecideaceae, Ascomycetes). Opera Botanica 110: 1-137.
Wirth V. 1995. Di2 Flechten Baden Wiirttembergs. Teil 1-2, Stuttgart, Ulmer.
MYCOTAXON
Volume 105, pp. 23-27 July-September 2008
Two new anamorphic rust fungi from northern areas of Pakistan
MUHAMMAD ASIM SULTAN’, IKRAM-UL-HAQ?, A.N. KHALID? & HAMID MUKHTAR?
asim418@ yahoo.com ‘Department of Biology, University College Lahore, Pakistan ikrhaq@ yahoo.com *Department of Botany, Government College University, Lahore, Pakistan
drankhalid@ yahoo.com *Department of Botany, University of the Punjab, Lahore, Pakistan
hamidwaseer@yahoo.com *Department of Botany, Government College University, Lahore, Pakistan
Abstract — Two anamorphic rust fungi, Aecidium pakistanicum on Spiraea tomentosa and Aecidium ikramii on Rosa sp. are described as new. These new discoveries raise the total of known rust taxa in northern Pakistan to forty-four.
Key words — Basidiomycota, new taxa, Phragmidium tuberculatum
Introduction
Northern areas of Pakistan are bounded by Afghanistan, People’s Republic of China, Indian-held territory of Kashmir and North West Frontier Province of Pakistan. The southern slopes near Kashmir lie within the Himalayan mountain system. Snow covered mountains with deep gorges and narrow valleys surround the area. Climatic conditions vary widely in the northern areas, ranging from monsoon influenced moist temperate zone in western Himalayas to arid and semi-arid cold desert in the northern Karakoram and Hindu Kush. Temperature in the valley bottoms can vary from extremes of 40°C in summer to less than 10°C in winter (Jacobose 1993).
The rust fungi of Northern areas of Pakistan are poorly known and only forty- two rust taxa have been published from the region so far (Ahmed 1956a, b;
* Author for correspondence
24 ... Sultan & al.
Gjaerum & Iqbal 1969, Kaneko 1993, Kakishima et al. 1993, Khalid et al. 1995, Khalid & Iqbal 1996a, b, 1997; Ahmed et al. 1997).
This paper reports two anamorphic rust fungi collected on rosaceous plants, Spiraea tomentosa L. and Rosa sp., from northern areas of Pakistan during 2001-02
Materials and methods
The host plants, Spiraea tomentosa and Rosa sp. were found infected with rust fungi. Aecidium pakistanicum on S. tomentosa from Fairy-meadows on July 22, 2001 and Aecidium ikramii on Rosa sp. from Bashu Jungle, Skardu, on October12, 2002 were collected by the senior author.
Dried herbarium materials were examined under light microscope and Scanning Electron Microscope (SEM). For LM observations, the hand sections and spores were mounted in a drop of Lactophenol solution on glass slides, gently heated to boiling point and then cooled. The measurements of spores are given in the form: min-max (mean = | standard division). The sections showing rust stages were observed under a microscope (NIKON YS 100) and micro photographed by Canon AE-1 programmed camera. For SEM the spores were attached to specimen stub in a drop of acetone and coated with carbon with Joel Jee-420 evaporator. The surface structure of spores was observed at 10 kV and photographed with Joel Jsn-6480 LV SEM.
Enumeration of taxa
Aecidium pakistanicum Sultan, I.U. Haq, Khalid & Mukhtar, sp. nov.
MycoBank MB 511527 (Plate 1, Figs A, C) Pycnia epiphylla, brunneis, dispersa, 0.1-0.2 mm. Aecidia hypophylla, dispersa, aggregate, revoluto, 0.2-0.7 mm, brunneis, peridio longo, hyalinae, cellula peridii ellipsoideae, 68-
88 x 15-19 um, pariete externo 3.5-5.5 um crasso, verrucoso, pariete interno 5.0-7.5 um crasso, verrucoso, Aecidiosporae 34-36 x 26-28 um, globosae vel subglobosae, ellipsoideae, hyalinae, brunneis, verruculoso, Poris obscures. Uredinia et telia ignota.
Holotype: On Spiraea tomentosa L., with 0 & 1 stages, Northern Pakistan, Fairy- Meadows, July 22, 2001, M.A. Sultan (LAH Herbarium # AS 41, Isotype GCU Herbarium).
EryMo.oey: Based on the name of country Pakistan.
Pycnia epiphyllous, brown, scattered, 0.1-0.2 mm. Aecia hypophyllous, scattered, sori appear aggregate, in groups, revolute, 0.2-0.7 mm, brown, peridia long, hyaline. Peridial cells (Fig 1A) ellipsoid, 68-88 x 15-19 um, outer wall 3.5- 5.5 um thick, verrucose, inner wall 5.0-7.5 um thick, verrucose. Aeciospores (Fig 1B) 34-36 x 26-28 um, globose to subglobose, ellipsoid, hyaline, brown, densely and finely verruculose, pores obscure. Uredinia and telia unknown.
Aecidium spp. nov. (Pakistan) ... 25
Plate 1. Figs. A-B: Aecidium pakistanicum on Spiraea tomentosa (A) Infected host plant Bar = 2.0 cm. (B) Aeciospores in Light microscope Bar =160 um. (C) Verruculose aeciospores in SEM. Bar = 18 um.
COMMENTS — Aecidium pakistanicum is unique among the known species of Aecidium in having a sac-like sorus and verruculose aeciospores. These characteristics separate A. pakistanicum from fourteen (Sato & Sato 1984) described aecium types. A. pakistanicum resembles none of the aecial types described so far.
Aecidium haqii Sultan, Khalid & Mukhtar, sp. nov. Plate 2, Figs. A-C MycoBANK MB 511528 Spermogonia, uredinia et telia ignota. Aecidia hypophylla, dispersa, aggregate, caeomoidea, paraphysibus peripheralis, flavida, 0.1-0.2 mm. Aecidiosporae globosae vel subglobosae, ellipsoideae, hyalinae, 20-26 x 19-25 um, clavo superne granulis refractivis, parietibus usque ad lum crassis, poris obscures.
Holotype: On Rosa sp., with I stage, Northern Pakistan, Bashu Jungle near Skardu, October 12, 2002, M.A. Sultan (LAH Herbarium # AS 39, Isotype GCU Herbarium)
26 ... Sultan & al.
Plate 2. Figs. A-C: Aecidium hagii on Rosa sp. (A) Infected host plant Bar = 2 cm. (B) Aeciospores in Light microscope Bar = 2 um. (C) Annulate aeciospores in SEM. Bar = 6.0 um
EtymMo oey: In honor of the renowned botanist Prof. Dr. Ikram-ul-Haq.
Spermogonia, uredinia and telia unknown. Aecia hypophyllous, scattered, in groups, caeomoid, paraphysis peripheral, yellow, 0.1-0.2 mm. Aeciospores (Fig 2) globose, subglobose, ellipsoid, hyaline, 20-26 x 19-25 um, annulate, wall 1 um thick, pores obscure.
ADDITIONAL SPECIMENS EXAMINED: LAH # 14131, 14132 S. Ahmed, on Rosa webbiana
Wall. ex Royle, Kaghan Valley August 28, 1949. Aecidium hagii was found on an unidentified Rosa species lacking thorns and with small leaves. The new species closely resembles Aecidium of Phragmidium tuberculatum J.H.H. Mill. Both have a caematoid aecidium and similarly sized aeciospores (see Wilson & Henderson 1966). Both species can be clearly differentiated on the basis of wall ornamentation of aeciospores. In P tuberculatum the aeciospores are verrucose and in Aecidium haqii aeciospores are annulate.
Aecidium spp. nov. (Pakistan) ... 27
Acknowledgements
We sincerely thank Dr. H. B. Gjaerum for help in verification of the taxa. We greatfully acknowledge Dr. Y. Ono, Ibaraki University, and Dr. S. H. Iqbal, University of Punjab, for reading the manuscript as pre-reviewers. We extend our thanks to A. R. Niazi for help in laboratory and to Drs. A. Latif, A. Hameed, M. Ghauri, Government College, University Lahore, for the SEM photographs. We are grateful to Dr. M. Weif’, Universitat Tubingen, for help in Latin diagnosis.
References
Ahmed S. 1956a. Uredinales of West Pakistan. Biologia 2(1): 29-101.
Ahmed S. 1956b. Fungi of Pakistan. Biological Society Pakistan Lahore Monograph 1: 1-126.
Ahmed S, Iqbal SH, Khalid AN. 1997. Fungi of Pakistan. Mycological Society of Pakistan,
_ Department of Botany, University of Punjab, Lahore, Pakistan.
Gjaerum HB, Iqbal SH. 1969. Some rust fungi from West Pakistan. Nytt Mag. fiir Botanik 16: 221-223.
Jacobose JP. 1993. Climatic records from Northern Areas of Pakistan. In: Culture Area Karakorum Newsletter, Tubingen, Germany, 3: 13-17.
Kaneko S. 1993. Parasitic fungi on woody plants from Pakistan. Cryptogamic Flora of Pakistan 2: 149-168.
Kakishima M, Okaen I, Ono Y. 1993. Rust Fungi (Uredinales) of Pakistan collected in 1991. Cryptogamic Flora of Pakistan 2: 169-179.
Khalid AN, Iqbal SH, Masood A. 1995. New records of Uredinales from Pakistan. Science International (Lahore) 7(4): 531-532.
Khalid AN, Iqbal SH. 1996a. Additions to the Rust Flora of Pakistan. Pak. J. Bot. 28(1): 114-117.
Khalid AN, Iqbal SH. 1996b. New Rusts from Pakistan. Can. J. Bot. 74: 506-508.
Khalid AN, Iqbal SH. 1997. Puccinia deosaiensis sp.nov on Epipactis helleborine (L.) Crantz. Can. J. Bot. 75: 864-886
Sato T, Sato S. 1984. Morphology of aecia of rust fungi. Trans. Br. mycol. Soc. 85: 223-238.
Sultan MA, Haq I, Khalid AN, Bajwa R. 2006. Some Uredinales from Northern areas of Pakistan. Pak. J. Bot. 38 (3): 837-841.
Sultan MA, Haq I, Khalid AN, Mukhtar H. 2007. A contribution to Uredinales of Northern Areas of Pakistan. Mycopath 4(2): 9-11.
Stewart RR. 1972. History and exploration of plants in Pakistan and adjoining areas. Flora of Pakistan, pp. 7.
Wilson M, Henderson DM. 1966. British Rust Fungi, Cambridge Univ. Press, Cambridge, U.K.
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MY COTAXON
Volume 105, pp. 29-36 July-September 2008
A new species of /saria isolated from an infected locust
*ZONGQI LIANG’, XUEYOU HE’, YANFENG HAN’, SHOUPING CAI’ & JIANGDONG LIANG!
zqliang1@yahoo.com.cn' swallow112886@yahoo.com.cn' ddl1977@yahoo.com.cn' ‘Institute of Fungus Resources, College of Life Sciences, Guizhou University Guiyang, China 550025 hexueyou@public.fz.fj.cn? caishouping@gmail.com? ?Fujian Academy of Forestry, Fuzhou, China 350012)
Abstract — Isaria locusticola sp. nov. was isolated from a locust in Fuzhou, Fujian Province, China. The new species is described and illustrated, and its relationships to the related species are briefly discussed. This fungus is characterized by the variability of the shape of conidia. Morphological characteristics and molecular analyses of ITS- 5.8S region sequences strongly support the establishment of I. locusticola in the genus Isaria.
Key Words — hyphomycetes, Paecilomyces, entomopathogenic fungi, morphology, rDNA analysis, Orthoptera
Introduction
Hodge et al. (2005) reviewed the nomenclatural status and history of the generic name Isaria and concluded that Isaria Pers. is a valid name. Consequently, they suggested that the name Isaria be used for those species previously assigned to Paecilomyces Bainier section Isarioidea Samson. Luangsa-ard et al. (2005) accepted this suggestion based on their molecular analysis and accepted 10 Paecilomyces species having Cordyceps teleomorphs as belonging in the genus Isaria. These species were I. farinosa (Holmsk.) Fr., I. amoenerosea Henn., I. cateniannulata (Z.Q. Liang) Samson & Hywel-Jones, I. cateniobliqua (Z.Q. Liang) Samson & Hywel-Jones, I. cicadae Migq., I. coleopterorum (Samson & H.C. Evans) Samson & Hywel-Jones, I. fumosorosea Wize, I. ghanensis (Samson & H.C. Evans) Samson & Hywel-Jones, I. javanica (Frieder. & Bally) Samson & Hywel-Jones and I. tenuipes Peck. They also suggested that both P ramosus Samson & H.C. Evans and P. xylariiformis (Lloyd) Samson belonged to Jsaria.
Corresponding author: Zonggqi Liang, zqliang1@yahoo.com.cn
30 ... Liang & al.
So far, more than 12 entomogenous species related to Paecilomyces have been assigned to Isaria.
Species of Paecilomyces are considered the most important pathogens parasitizing lepidoptoran insects (Evans & Samson 1982, Kubatova & Dvorak 2005). Species that parasitize orthopteran insects, however, have been rarely reported. Evans was the first to isolate fungus P. reniformis Samson & H.C.Evans from an infected grasshopper (Tettigoniidae, Orthoptera) (Samson 1974); the same fungus has been also recently isolated from long horned grasshoppers (Tettigoniidae, Orthoptera) in Sulawesi, Indonesia (Kalkar et al. 2006). Liang et al. (1991) described Paecilomyces atrovirens Z. Q. Liang & A.Y. Liu from an infected Tettigonia from Guizhou Province, China. During recent research on Chinese entomogenous fungi, the authors discovered an undescribed Isaria species from Fujian Province (eastern China), which is described below.
Materials and methods
Sample collection and strain isolation
Strain GZUIFR-Fj0916 was isolated from an infected adult of Chondracris sp. (Acrididae, Orthoptera) collected in the Fuzhou National Forest Park, Fujian Province. The infected insect collected from the field was first removed soil and other debris. Few conidia were transferred to plates of Martins medium. Cultures were incubated at 26°C until colonies were developed. Pure cultures having uniform conidiogenous structures from the infected insect were transferred to Sabouraud’s slants and stored in the Institute of Fungus Resources, Guizhou University.
Strain identification
The strain was transplanted to Czapek agar and potato dextrose agar (PDA), incubated at 26°C for 14 days, and identified based on colony characters, conidiogenous structures, temperature responses following Brown & Smith (1957) and Samson (1974) and molecular analysis.
DNA extraction |
Taq enzyme and dNTP were supplied by Shanghai Sangon. ‘The strain GZUIFR- Fj0916 used for the molecular identification was incubated on Czapek agar. DNA was extracted from the fresh sporulating cultures following Tigano- Milani et al. (1995). The extracted DNA was stored at -20°C.
PCR and ITS rDNA sequencing
The internal transcribed spacer (ITS) region including the 5.88 rDNA was amplified - by polymerase chain reaction (PCR) using the primers ITS5 (5’- GGT GAG AGA TTT CIG TGC -3’).and IT$4,(5-TGC TCC GCT TAT TGA TAT GG-3’). After a first denaturation step at 94 °C for 5 min, the amplification reaction was performed for 35 cycles with denaturation at 94°C for 40 s, annealing at 49°C for 40 s, and extension at
Isaria locusticola sp. nov. (China) ... 31
Table 1 Fungi used in the study with their GenBank accession numbers
NAMES GENBANKNo NAME GENBANK No. Byssochlamys fulva AY753341 I, javanica AY624186 B. nivea AY753339 I, javanica DQ403723 Cookeina colensoi AF394531 I. locusticola EU363501 C. venezuelae AF394041 Paecilomyces aerugineus AY753346 Cordyceps militaris AY725790 P. amoenerosea AY624169 C. takaomontana AY624198 P. carneus AY624170 Isaria japonica AF200370 P. marquandii AB099511 I. japonica AY624199 P. militaris EU363502 I. cateniannulata AF368802 P. niphetodes AY624192 I. cateniannulata AY624172 P. penicillatus AY624194 I. cateniobliqua AF368799 P. purpureus EF640809 I. farinosa AY624179 P. reniformis ARSEF429 I. fumosorosea AJ608982 P. reniformis ARSEF577 I. ghanensis AY624185 P. reniformis DQ069283 I. cateniobliqua AY624173 I. tenuipes AF368808 I. cicadae AB085887 P. variotii AY373941 _I. coleopterorum AY624176 P. viridis AY624197
72°C for 1 min; followed by a final extension step at 72°C for 10 min. PCR products were purified using the Agarose Gel DNA Purification kit version 2.0 (Takara Company) according to the manufacturer's protocol and were sequenced with the above primers at Beijing Sunbiotech Co. Ltd. The sequence of ITS1-5.8S-ITS2 rDNA region of strain GZUIFR-Fj0916 was submitted to GenBank (accession number: EU363501).
Sequence alignment and phylogenetic analysis
Table 1 lists strains used in the molecular study. Some ITS1-5.8S-ITS2 region nucleotide sequences of representative Isaria and Paecilomyces species were obtained from GenBank. The sequence of the new species was aligned with related fungi using the Clustal X1.83 computer program for multiple sequence alignment and manually corrected. The phylogenetic tree was constructed by neighbor-joining method (NJ) of MEGA version 3.1 (Kumar et al. 2004). Confidence values for individual branches were determined by bootstrap analysis (1000 replications).
Results and discussion Taxonomy
Isaria locusticola Z.Q. Liang, X.Y. He & Y.F. Han, sp. nov. Fig. 1 MycoBank MB 511579; GENBANK EU363501
In agaro Czapekii, coloniae 40 mm diam, 14 diebus ad 26°C, ablae ad luteolum; reversum luteolum. Hyphae septatae, hyalinae. Conidiophora erecta, septata, hyalina. Phialides 4.2-4.8 x 1.2-2 um, e basi inflata ellipsoida, claviformisa vel cylindricusa in collum longum angustatae. Conidia continua, hyalina, levia, ellipsoidea, (2.4—)3-3.6(-4.2) x (1.2-)1.5-1.8(-2) um, vel 4.8 x 3 um; fusiformia vel cylindricusa 3.5-4.5 x 1.2-1.5 um. Teleomorphosis ignota.
32 ... Liang & al.
Fig. 1 The host insect and conidiogenous structures of I. locusticola. 1. A locust infected by I. locusticola; 2-5. The conidiogenous structures; 6. Conidia.
Holotypus GZUIFR-Fj016 isolatus, e Chondracris sp.(Acrididae, Orthoptera), Fuzhou regio,
Fujian provinca, China. IX, 2007, X.Y. HE et S. P CAL in Guizhou Univ, conservatur. Cotony on Czapek agar, attaining a diameter of 40 mm within 14 days at 26°C, flat, white to yellowish, villiform. Reverse yellowish. VEGETATIVE HYPHAE hyaline, smooth-walled. CONIDIOPHORES septate, forming branches with
Isaria locusticola sp. nov. (China) ... 33
Table 2. A comparison of morphological characters between I. locusticola and related species
Names Colony Conidia Hosts Ff ihe, iP dab orcen cylindrical, pee sp. 4.2-7.2 x 2.4-3um (Acrididae, Orthoptera)
cylindrical to fusiform
Li : hit ; javanica white 57 ae 417 in lepidopteran insects I. locusticola white-yellowish ecae Sh etligs ae ee 2.4-4.2 x 1.2-2 um (Acrididae, Orthoptera)
fusiform to oblong-ellipsoid 3.9-4.9 X1.2-1.5, 1m
greyish-green, — reniform Grasshopper
P. reniformis ‘ eh ais olive-green 4.5-5.5 x 2-3.5 um (Tettigontidae, Orthoptera)
phialides in whorls of 2 to 4. PHIALIDEs divergent, 4.2-4.8 x 1.2-2 um, consisting of an ellipsoidal, cylindrical or clavate basal portion and a distinct thin neck. Conrp1A hyaline, smooth-walled, variable, ellipsoid, (2.4-)3-3.6(—4.2) x (1.2-) 1.5-1.8(-2) um, a few up to 4.8 x 3 um; fusiform to oblong-ellipsoid, 3.5-4.5 x 1.2-1.5 um. Bi-celled rare.
TELEOMORPH: unknown.
DISTRIBUTION: Fujian Province, China. MATERIAL EXAMINED: The holotype, GZUIFR-Fj0916, was isolated by X.Y He & S.P. Cai from an infected cotton grasshopper Chondracris sp. (Acrididae, Orthoptera), Fuzhou, Fujian Province, China, September, 2007, deposited in the Institute of Fungus Resources, Guizhou University. The two species of Paecilomyces known to infect locusts — P. reniformis and P. atrovirens (Samson 1974, Liang et al. 1991, Kalkar et al. 2006) — distinctly differ from I. locusticola (Table 2). P reniformis produces grayish-green or olive-green colonies and reniform conidia, while Isaria locusticola produces white colonies and ellipsoid to oblong-ellipsoid or fusiform conidia. Some interesting bioassay tests also show that P reniformis isolated from grasshoppers in Indonesia did not infect lepidopteran larvae at the concentrations tested and the exposure method used, although the species did infect long- and short- horned tettigoniid grasshoppers, suggesting that P reniformis may be specific to this insect group (Kalkar et al. 2006).
Paecilomyces atrovirens differs from I. locusticola by its dark green colony, long cylindrical conidia (4.2-7.2 um), phialides with a lecythiform basal portion and imbricate conidial chain (Liang et al. 1991). I. locusticola and I. javanica have similar conidiogenous structures , but the conidia of the former are ellipsoid,(1.2-2 x 2.4-4.2 um), fusiform to oblong- ellipsoid (3.5-4.5 x 1.2-1.5 um), while they are cylindric (5-7.4 x 1.5-1.7 um) in the latter (Samson
34 ... Liang & al.
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Tsciria tenuipes AP 368808 * Cordyceps takaomontana AY624198 L. japonicadF 200370 id os BL japonicaAY624199 1. ghanensisAY624185 é 1. fumosoroseasJ6038982 I, coleopterorumAY624176 1, civcadaeABO8588? I. cateniannulataAF 368802 88 § F cateniannulataAY624 172 ag jPaecilomyces militariskU363502 C. militar isAY7 25790 1 farinosaAY624179 P. reniformisDQ069283 P, reniformisARSEFS?7 P. reniformisARSEF429 eg [4 Javanicad Y624186 L javanicaDQ 403723 f locusticolakU 363501 L, amaeneraseaAY624169 1. cateniabliquaAY624173 I. cateniobliquadF 368799 P, niphetodes AY624 192 P. penicillatusAY624 194 P. carneusAY624170 P. purpureusEF 640809 P, viridisAY624197 P. marquandiABO995 11 P. aerugineusAY 733346 P. var totiiaY 373941 Byssochlamys filva A¥753341
Il-2 100
my
[l-2-1
36
-1
Byssochlamys ntvea AY733339
Cookeina colensoidF 394531
150 Cookeina venezuelaedF 394041
Fig. 2 Phylogenetic tree based on analysis of rDNA ITS1-5.8S-ITS2 sequences of I. locusticola
and some related species.
1974). A morphological comparison of I. locusticola and related species is
shown in Table 2.
Molecular identification
A BLAST search through GenBank was performed by using the ITS sequence of I. locusticola as the query. Close matches showing maximal sequence identities of” 92-98% included I. javanica, I. fumosorosea, I. cateniannulata, I. cateniobliqua,
I. amoenerosea, and I. farinosa. The ITS sequences of these species and other
related species of Paecilomyces and other entomogenous fungi were retrieved
from GenBank for phylogenetic analysis.
Isaria locusticola sp. nov. (China) ... 35
Relationships of I. locusticola and related species in Isaria and Paecilomyces were showed in the phylogenetic tree based on analysis of rDNA ITS1- 5.8S - ITS2 sequences (Fig. 2). Cookeina colensoi and C. venezuelae were designated as outgroups.
Luangsa-ard et al. (2004, 2005) demonstrated that Paecilomyces is polyphyletic, with species falling in two subclasses, Sordariomycetidae and Eurotimycetidae. Two clades (Clades I and II) were recognized in the phylogenetic tree. Clade I consisted of the thermophilic Byssochlamys fulva, B. nivea, P. aerugineus and P. variotii in the Eurotiales (Eurotimycetidae). Clade II consisted of Paecilomyces spp. (Hypocreaceae, Hypocreales, Eurotimycetidae) and Isaria spp. (Clavicipitaceae, Hypocreales, Eurotimycetidae,). I. locusticola grouped with I. javanica as sister to I. cateniobliqua and I. amoenerosea. ‘This association was well supported by bootstrap analysis (values = 96%).
Luangsa-ard et al. (2005) did not include P. reniformis in their phylogenetic analyses. Our molecular analyses suggest that P reniformis does not group with other Paecilomyces species but rather belongs to the entomogenous Isaria group, which further suggests that P reniformis belongs to Isaria, not Paecilomyces. Although both P reniformis and I. locusticola parasitize locusts, their separation in the tree support them as independent species.
Both I. cateniobliqua and I. amoenerosea, which are characterized by red colonies, are in the same subclade but apparently differ from the subclade of I. locusticola and I. javanica. Although I. locusticola and I. javanica are closely related by molecular analysis, they produce different conidia and live on different insect hosts. Both the phenotypic description and molecular analyses support recognition of strain GZUIFR-Fj0916 as a new member of Isaria.
Acknowledgements
This study was supported by the National Natural Science Foundation of China (No. 30499340). We are grateful to Prof. Y.C. Dai and Dr. R.T.V. Fox for their comments on the manuscript. We also warmly thank Dr. $.R. Pennycook and Dr. L.L. Norvell for editorial review and revisions.
References
Brown AHS, Smith G. 1957. The genus Paecilomyces Bainier and its perfect stage Byssochlamys Westling. Trans Brit Mycol Soc. 40(1): 17- 89.
Evans HC, Samson RA. 1982. Cordyceps species and their anamorphs pathogenic on ants (Formicidae) in tropical forest ecosystem I. The Cephalotes (Myrmicinae) complex. Trans Brit Mycol Soc. 79: 431- 453.
Hodge KT, Gams W, Samson RA, Korf RP, Seifert KA. 2005. Lectotypification and status of Isaria Pers.:Fr. Taxon 54 (2): 485-489.
Kalkar O, Carner GR, Scharf D, Boucias DG. 2006. Characterization of an Indonesian isolate of Paecilomyces reniformis. Mycopathologia 161: 109-118.
36 ... Liang & al.
Kubatova A, Dvorak L. 2005. Entomopathogenic fungi associated with insect hibernating in underground shelters. Czech Mycol. 57(3-4): 221-237.
Kumar S, Tamura K, Nei M. 2004. MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Briefings in Bioinformatics 5: 150-163.
Liang ZQ, Liu AY, Feng DM. 1991. Some entomogenous fungi from Fanjing Mountain Preserve in China. Acta Mycologica Sinica 12 (2): 110 -117.
Luangsa-ard JJ, Hywel-Jones NL, Manoch L, Samson RA. 2005. On the relationships of Paecilomyces sect. Isarioidea species. Mycol. Res. 109(5): 581-589.
Luangsa-ard JJ, Hywel-Jones NL, Samson RA. 2004. The polyphyletic nature of Paecilomyces sensu lato based on 18S-generated rDNA phylogeny. Mycologia 96 (4): 773-780.
Samson RA. 1974. Paecilomyces and some allied Hyphomycetes. Studies in Mycology 6: 1-119.
Tigano-Milani MS, Samson RA, Martins I, Sobral BWS. 1995. DNA markers for diferentiating isolates of Paecilomyces lilacinus. Microbiology 141: 239-245.
MYCOTAXON Volume 105, pp. 37-42 July-September 2008
Calathella columbiana (Basidiomycota): new record of a cyphelloid fungus from Brazil
MARCELO A. SULZBACHER’, DENNIS E. DESJARDIN? & JAIR PUTZKE!
marcelo_sulzbacher@yahoo.com.br 'Universidade de Santa Cruz do Sul, Laboratorio de Botanica Av. Independeéncia, 2293, Santa Cruz do Sul, RS, BRAZIL
ded@sfsu.edu *San Francisco State University, Department of Biology 1600 Holloway Ave., San Francisco, CA, 94132, USA
Abstract — Calathella columbiana is described and illustrated based on material collected in Santa Catarina State, southern Brazil. The species was originally described from Colombia in 1983 and no more recorded from South America, the present record is the second one for the species in this continent.
Key words — Agaricomycetidae, Cyphellaceae s.l., neotropical fungi
Introduction
Calathella D.A. Reid is a small genus of tubular or cupulate cyphelloid fungi described by Reid (1964). Agerer (1983) studied six Calathella species and considered the following characters to distinguish it from other related genera: incrusted surface hairs with rounded tips, suburniform basidia, and oblong- elliptical to cylindrical basidiospores. Currently, eight species are known in the genus (Kirk et al. 2001).
From Brazil, there are few reports on cyphelloid mushrooms (Viégas 1945, Putzke 2002), possibly because of their much reduced size and difficult taxonomy. The genus Calathella was only recently reported in Brazil by de Meijer (2001, 2006), who mentioned unidentified specimens growing on Cyathea tree-ferns.
In this paper, we report the occurrence of Calathella columbiana, the first species reported from Brazil, collected in montane vegetation of Santa Catarina State.
38 ... Sulzbacher, Desjardin & Putzke
Materials and methods
Specimens were collected and photographed, then macroscopically analyzed following Bodensteiner et al. (2001). Microscopical observations were made from material mounted in 3% KOH and Melzer’s reagent. Measurements of the basidiospores do not include the apiculus; the shape was determined based on Q (length-width ratio) values according to Largent et al. (1977). Color codes used in the description of the species are those from Maerz & Paul (1930). Specimens are deposited in the herbarium SMDB.
Taxonomy
Calathella columbiana Agerer, Mitt. Bot. Staatssamml. Miinchen 19: 192, 1983. Fics. 1-4
BASIDIOMES 0.5—0.7 mm diam., scattered to clustered, with a short, stalk-like, attenuated base when young, cupulate, unlobed but some becoming lobed in age, with 2—5 broad finger-like lobes; margins slightly involute; external surface woolly, pale-brown, becoming nearly white at the margin. HYMENIUM smooth, glabrous, cream-colored. SuBicuLUM lacking. EXTERNAL SURFACE HAIRS 2—4 um diam., somewhat twisted, with obtusely rounded tips; tips hyaline, grading into pale yellowish toward the base from intraparietal pigmentation; thickness and color of adjacent trama hyphae similar to those of the bases of external hairs; hairs with irregular incrustation patterns, with alternating densely incrusted and nearly crystal-free areas, and tips sometimes loosely incrusted or entirely glabrous; hairs not swelling in 5 % KOH, dextrinoid. TRAMA neither agglutinated nor gelatinous; hyphae 2.5—4 um diam, with walls 0.5 um thick; outer tramal elements +/- parallel to basidiome surface, frequently septate, yellowish from intraparietal pigmentation; SUBHYMENIAL HYPHAE hyaline, short-celled, hardly differentiated from inner tramal hyphae. CLAMP CONNECTIONS absent. Bastp1A 24—27 x 6—7 um, clavate, 4-spored; sterigmata narrowly horn-like; lacking distinctive transition elements between basidia and surface hairs at basidiome margin. BASIDIOLEs (17—) 19-27 (—32) x (3.5-) 5—7.5 um, clavate. CysTrp1a absent. BASIDIOSPORES 7.5—10 x (-3) 4—5 um [x = 8.4+0.7 x 4.1 + 0.6 um, Q= 1.7 - 2.7,Q_ =2.4 + 0.6, n = 20], asymmetrically oblong-elliptical to cylindrical or slightly allantoid, apiculate, smooth, hyaline, thin-walled, inamyloid, nondextrinoid.
HABITAT: scattered to gregarious on the abaxial surface of decayed grass leaves _ Poaceae, in cleared forest. STUDIED MATERIAL: BRAZIL. Santa Catarina State, municipality of Riqueza, Linha Alta
Riqueza, (27° 02°01,4”S — 53°20°13,2”W), 340 m alt., 27.XII.2006, leg. A.A. Spielmann & M.A. Sulzbacher-89 (SMDB 10.988)
Calathella columbiana in Brazil ... 39
Fig. 1. Calathella columbiana. Fresh basidiomes.
REMARKS: The studied Brazilian material is similar to C. digitiformis Bodenst. et al. (Bodensteiner et al. 2001), another recently described species from Bali, Indonesia. However, the latter species presents a set of features that differ from C. columbiana,; i.e., digitiform basidiomes 0.7—1.5 mm diam. with numerous finger-like lobes, narrower basidiospores (2.5—3.5 um diam. with mean width 3 um; Q_ = 2.8) and smaller basidia (14.5—21 x 4.5-6 um — Bodensteiner et al. 2001). Both species grow on herbaceous substrates.
The Brazilian collection exhibits intermediate characters between the species cited above. For example, the size of the basidiospores and basidia are smaller than those described for C. columbiana by Agerer (1983). In addition, a few basidiomes are slightly lobed, but not as dissected as those of C. digitiformis. Based on the neotropical distribution and macroscopic features, we consider our specimen within the morphological variation of C. columbiana, which is a poorly known species. Prior to this report C. columbiana was known only from Colombia (type locality) and its distribution is now extended to southern Brazil.
AO ... Sulzbacher, Desjardin & Putzke
Fig. 2-4. Calathella columbiana: 2) Basidiospores. 3) Basidia. 4) Basidiome section.
The taxonomic circumscription and phylogenetic position of the genus Calathella is somewhat controversial, and recent molecular studies indicate unresolved phylogenetic relationships suggesting that Calathella is not monophyletic. Calathella columbiana was segregated from the other Calathella species included in the analyses of Bodensteiner et al. 2004. In this work, C. columbiana was sister of Mycenoporella lutea Overeem, a mycenoid species with a poroid hymenophore and together they were sister of the Entolomataceae. In comparison, C. mangrovei E.B.G. Jones & Agerer, and C. gayana (Lév.) Agerer were distantly related to each other in the Nia clade and collectively were sister of the Omphalotaceae. It should be noted, however, that the relationships amongst the major clades presented in Bodensteiner et al. (2004) were significantly different from those in the multi-locus analyses of Matheny et al. (2006). Singer (1986) accepted Calathella as a synonym of the genus Flagelloscypha because both taxa formed cyphelloid basidiomes covered with dextrinoid, thick-walled hairs, and he maintained the distinction from the genus Lachnella whose members have inamyloid hairs. In addition Singer (1986) was
Calathella columbiana in Brazil ... 41
reluctant to accept the numerous segregate genera recognized by Agerer (1983) that were delimited by subtle micromorphological features. Recent molecular data (Bodensteiner et al. 2004; Binder et al. 2005) support the acceptance of Calathella, Flagelloscypha and Lachnella as distinct genera, but the relationships amongst them are equivocal, and as already indicated, Calathella is polyphyletic. It should be noted that the type species of Calathella, C. eruciformis (Batsch) D.A. Reid, is not included in any published molecular studies, and accordingly whether its phylogenetic affinities lie with C. columbiana or C. mangrovei or C. gayana cannot be ascertained. Until the generic circumscription of Calathella is clarified, we tentatively accept C. columbiana in Calathella.
Acknowledgments
The authors thank to Adriano A. Spielmann for the photograph of the collection. To Vagner G. Cortez and Gilberto Coelho for providing the line drawings. To Dr. Timothy J. Baroni (State University of New York - College at Cortland) and Dr. D. Jean Lodge (Center for Forest Mycology Research, USDA Forest Service) for kindly reviewing the manuscript.
Literature cited
Agerer R. 1983. Typusstudien an Cyphelloiden Pilzen IV. Lachnella Fr. s.1. Mitt. Bot. Staatssamml Miinchen 19: 163-334.
Binder M, Hibbett DS, Larsson K-L, Larsson E, Langer E, Langer G. 2005. The phylogenetic distribution of resupinate forms across the major clades of mushroom-forming fungi (Homobasidiomycetes). Syst. Biodiv. 3: 113-157.
Bodensteiner P, Agerer R, Desjardin DE, Horak E. 2001. A new species of Calathella from Bali. Mycologia 93: 1010-1013.
Bodensteiner P, Binder M, Moncalvo JM, Agerer R, Hibbett DS. 2004. Phylogenetic relationships of cyphelloid homobasidiomycetes. Mol. Phylogen. Evol. 33: 501-515.
de Meijer AAR. 2001. Mycological work in the Brazilian state of Paranda. Nova Hedwigia 72: 105-159.
de Meijer AAR. 2006. Preliminary list of the macromycetes from the Brazilian state of Parana. Bol. Mus. Bot. Munic., Curitiba 68: 1-55.
Kirk PM., Cannon PE, David JC, Stalpers JA. 2001. Ainsworth & Bisby’s Dictionary of the Fungi. 9" ed. Wallingford (UK): CABI.
Largent DL, Jonhson D, Watling R. 1977. How to Identify Mushrooms to Genus. III. Microscopic features. Eureka (USA): Eureka Publishing.
Maerz AJ, Paul MR. 1930. A dictionary of color. New York (USA): McGraw-Hill.
Matheny PB, Curtis JM, Hofstetter V, Aime MC, Moncalvo J-M, Ge Z-W, Yang Z-L, Slot JC, Ammirati JF, Baroni TJ, Bougher NL, Hughes KW, Lodge DJ, Kerrigan RW, Seidl MT, Aanen DK, DeNitis M, Daniele GM, Desjardin DE, Kropp BR, Norvell LL, Parker A, Vellinga EC, Vilgalys R, Hibbett DS. 2006. Major clades of Agaricales: a multilocus phylogenetic overview, Mycologia 98: 982-995.
Putzke J. 2002. Agaricales (Fungos - Basidiomycota) pleurotdides no Rio Grande do Sul. I - Anthracophyllum, Aphyllotus, Campanella, Chaetocalathus e Cheimonophyllum. Cad. Pesq., Sér. Biol. 14: 45-66.
42 ... Sulzbacher, Desjardin & Putzke
Reid DA. 1964. Notes on some fungi of Michigan. I. “Cyphellaceae’. Persoonia 3: 97-154. Singer R. 1986. The Agaricales in Modern Taxonomy. 4th ed. Koenigstein (Germany): Koeltz
Scientific Books.. Viégas AP. 1945. Alguns fungos do Brasil, VII-VIII: Cyphellaceae e Thelephoraceae. Bragantia >:
253-290.
MYCOTAXON
Volume 105, pp. 43-52 July-September 2008
A new species within the Gymnopus dryophilus complex (Agaricomycetes, Basidiomycota) from Italy
ALFREDO VIZZINI', GIOVANNI CONSIGLIO’, VLADIMIR ANTONIN? & MARCO ContTU*
‘alfredo. vizzini@unito.it Dipartimento di Biologia Vegetale, Universita di Torino Viale Mattioli 25, 10125 Torino, Italy *giovanni.consiglio@fastwebnet. it Via Ronzani, 61 - I 40033 Casalecchio di Reno, Bologna, Italy -vantonin@mzm.cz
Moravian Museum, Department of Botany, Zelny trh 6
CZ-659 37 Brno, Czech Republic
* marcocontu@interfree.it Via Traversa via Roma, 12 (I Gioielli 2) - 07026 Olbia, Sassari, Italy
Abstract — Gymnopus inexpectatus, collected from a coastal ecosystem in Italy on a dead branch of Quercus ilex, is proposed as a new species. ITS and LSU rDNA sequences place it in the G. dryophilus complex within sect. Levipedes subsect. Levipedes, where its compact coralloid pileipellis structure and broadly ellipsoid spores (Q = 1.5) make it unique. A Latin diagnosis, technical description, illustrations, and taxonomic speculations on Gymnopus-Marasmiellus relationships are provided. Comparisons with the European, North/South American and Indonesian species that represent the most closely related species are also presented.
Key words — Collybia, gymnopoid/omphalotoid clade, Mediterranean Agaricales, Omphalotaceae, taxonomy
Introduction
The genus Gymnopus (Pers.) Roussel (Agaricomycetes, Agaricales, Omphalotaceae) includes saprotrophic (mainly litter decay) non-mycotrophic agarics (Antonin & Noordeloos 1997, 2008) with white to cream-coloured spore-prints, stipes that are not insititious but usually with basal mycelium and/ or rhizomorphs, rooting or rarely originating from a sclerotium, a cutis type pileipellis composed of narrow, cylindrical hyphae or a complex structure of inflated terminal elements (dryophila—structure), inamyloid, non-dextrinoid,
44 ... Vizzini & al.
acyanophilic basidiospores, and non-dextrinoid contextual hyphae. Gymnopus is a character-poor genus, since many taxa diverge in only very subtle features, such as basidiomata macromorphology, pileipellis structure, cystidia shape and basidiospore size. This complicates species identification, often daunting from a morphological perspective.
While studying mycobiota of the Natural Park of Rimigliano (S. Vincenzo, Livorno, Tuscany, Italy), 120 hectares of sandy coastal ecosystem formed by holm oak (Quercus ilex L.) and cork oak (Q. suber L.) woods and by a dense mediterranean pine forest (Pinus pinea L., P. pinaster Aiton), specimens of an unusual Gymnopus species have been found. After comparison with species from Europe, (e.g. Vilgalys & Miller 1987a, Noordeloos 1995, Bon 1999, Antonin & Noordeloos 1997, 2008), North and South America (e.g. Vilgalys & Miller 1983, Halling 1983, 1996, 1997; Mata et al. 2006) and Asia (Indonesia, Wilson et al. 2004), it is considered to represent a new species. It is described and illustrated below.
Materials and methods
Macro- and micromorphology
Macroscopic characters were examined from fresh material. Microscopical studies are based on dried material using a Leica DM 4500 B and an Olympus BX50 light microscope with magnifications up to 1000 x. Observations were made on mounts in the following reagents: Congo Red in 10% ammonia, Cresyl-Blue in water, Melzer’s reagent and 3% KOH. Measurements are based on observation of 30 basidiospores of two basidiomata (apiculus not included). The following abbreviations have been used: E = the quotient of length and width of the spores; Q = the mean value of E values in all collections studied; L = number of entire lamellae; | = number of lamellulae between each pair of entire lamellae. All examined material has been deposited and preserved in MCVE (Museo Civico di Storia Naturale, Venice, Italy). Herbarium abbreviations follow Holmgren & Holmgren (1998). Latin description of the new species has been deposited in MycoBank.
DNA amplification and sequencing
The DNA from a dried basidiome was extracted using the DNA Plant Minikit (Qiagen) and following manufactor’s instructions. The partial internal transcribed rDNA spacer (ITS) and the D2 region of the large subunit (LSU) rDNA were amplified through the polymerase chain reaction (PCR). The — primer pairs ITS1F/ITS4 for ITS (White et al. 1990, Gardes & Bruns 1993) and 0061/NDL22 for LSU (van Tuinen et al. 1998, Kjoller & Rosendahl 2000) were used both for PCR reactions and sequencing. The PCR products were purified
Gymnopus inexpectatus sp. nov. (Italy) ... 45
by the PCR purification kit and sequenced by DINAMYCODE S.r.l. Searches for similar sequences allowing taxonomic identification were conducted using the BLASTN algorithm available through the NCBI (http://www.ncbi.nlm. nih.gov/blast/Blast.cgi). The two consensus sequences have been deposited in GenBank and the relative accession numbers are listed with the collection.
Taxonomy
Gymnopus inexpectatus Consiglio, Vizzini, Antonin & Contu, sp. nov. FIGURES 1,2 MycoBank MB 511701
Pileus 25-30 mm latus, convexus vel plano-convexus, hygrophanus, ochraceo-brunneus, maculis ferruginosis designatus, laevis, glaber. Lamellae (sub)distantes, L = 25-30, 1 = 2-6 (7), adnatae, cremeo-roseae. Stipes longus tam quam pilei diametron vel vix longior, 5-6 mm crassus, cylindraceus, insiticius, concolor cum pileo, longitudinaliter fibrillosus, basi tomentosus. Caro cartilaginea. Odor nullus. Sapor mitis. Sporae 5-6 (7.0) x 3.2-4 (4.5) um, E = 1.4-1.7, (late) ellipsoideae, ovoideae, tenuitunicatae, hyalinae, laeves, inamyloideae. Basidia tetraspora, clavata. Cheilocystidia 27-45 x 5-8 um, lageniformia, subfusoidea vel subcylindracea, saepe rostrata, saepe irregularia vel subcoralloidea. Lamellarum trama ex cylindraceis vel fusoideis hyphis constituta. Pileipellis ex singulari specie “dryophila-structurae” efformata, ex versiformibus, cylindraceis, clavatis, fusoideis vel utriformibus, hyalinis vel pigmento parietali luteolo-brunneolo hyphis cum digitiformibus propagationibus, constituta. Caulocystidia discretis cystidiis vel (plerumque) terminalibus cellulis similia. Fibulae adsunt ubique.
Holotypus in loco Rimigliano dicto prope S. Vincenzo (Livorno, Italy), 2.XII.2006 lectus, leg. G. Consiglio, E. Franceschini et G. Perdisa, in Herbario MCVE sub n° 16126 conservatur.
ETryMoLocy: from inexpectatus (Latin), meaning “unexpected, unforeseen’.
PILEUS 25-30 mm, convex to plano-convex, applanate with age, with slightly involute margin, hygrophanous, not translucently striate when moist, brownish ochre, rusty red spotted, pallescent to ochraceous cream, smooth, glabrous, slightly tomentose at margin. LAMELLAE rather distant, subdistant, L = 25-30, | = 2-6 (7), adnate, rosy cream, with entire to hardly uneven, concolorous edge. STIPE of the same length, or slightly longer, than pileus diameter, 5-6 mm thick, cylindrical, slightly broadened at base, insititious, stuffed, concolorous with the pileus, longitudinally fibrillose, tomentose at base, without rhizomorphs. ConrTEXT cartilaginous, concolorous with the external surfaces. SMELL none, indistinct. TasTE mild.
BASIDIOSPORES 5-6 (7) x 3.2-4 (4.5) um, on average 5.9 x 3.8 um, E = 1.4-1.7, Q = 1.5, (broadly) ellipsoid, ovoid, smooth, thin-walled, hyaline, inamyloid (Fig. 2b). Basrp1a 20-25 x 6-7.5 um, 4-spored, clavate, often subcapitate. BASIDIOLES 14-23 x 3-6 um, clavate, cylindrical. CHEILOCYSTIDIA 27-45 x 5—- 8 um, lageniform, subfusoid or subcylindrical, often rostrate, often irregular to subcoralloid, thin-walled (Fig. 2a). PLEUROCYSTIDIA absent. HYMENOPHORAL TRAMA consisting of cylindrical to fusoid, thin-walled, up to 13 um wide hyphae. PILEIPELLIS a special form of “dryophila-structure’, composed of versiform,
46 ... Vizzini & al.
Fig. 1. Gymnopus inexpectatus (holotype). Basidiomata. Scale bar = 10 mm.
cylindrical, clavate, fusoid or utriform, + thin-walled, 30-62 x 8-15 um elements, which are irregular, coralloid or with + digitate projections, hyaline to pale yellowish-brownish in KOH (Fig. 2d); with parietal and fine epiparietal- encrusting pigment. STIPITIPELLIS a cutis consisting of cylindrical, parallel, thick-walled (up to 1 um), smooth, up to 7 um wide hyphae. CAULOCYSTIDIA in the form of discrete cystidia or (mostly) terminal cells, 17-48 x 4.5-10 um, adpressed to erect, cylindrical, clavate, regular, irregular to coralloid, + thin- to thick-walled (Fig. 2c). CLAMP CONNECTIONS present in all tissues. CHEMICAL REACTIONS, no part of the basidiome dextrinoid or amyloid and not staining green in KOH.
HasitTat. On a little branch of Quercus ilex fallen on the soil.
COLLECTION EXAMINED — Italy, S. Vincenzo, Livorno, in the Quercus ilex forest of Rimigliano: two basidiomes on a branch of Q. ilex, 2°! Dec. 2006, leg. G. Consiglio, E. Franceschini et G. Perdisa (holotype MCVE 16126; GenBank accession numbers EU622905 and EU622906).
Molecular results
The ITS and LSU rDNA D2 regions were amplified from the dried basidiome. The size of the ITS and LSU rDNA fragments was 238 bp and 606 bp, respectively. Searching for similar sequences in GenBank database pointed to taxa within the Gymnopus dryophilus complex (Tab. 1).
Gymnopus inexpectatus sp. noy. (Italy) ... 47
Fig. 2. Gymnopus inexpectatus. Micromorphologic characters of the basidiomata. a. Cheilocystidia. b. Basidiospores. c. Caulocystidia. d. Pileipellis elements (all from holotype). Scale bar = 20 um.
Discussion
This new species is well characterised by a unique combination of features. Nevertheless its taxonomic relationships are problematic: our collection combines characters such as a gymnopoid habitus (viz. pileus relatively fleshy, not distinctly striate, lamellae usually not distant, stipe not filiform to hair- like but cartilaginous), that suggest a close relationship with Gymnopus, with characters (viz., coralloid pileipellis structure, lignicolous habit, insititious stipe) reminiscent of several Marasmiellus s.]. species (Antonin et al. 1997, Antonin & Noordeloos 1993, 1997, 2008). However, recent molecular analyses by Mata et al. (2004, 2006) and Wilson & Desjardin (2005) note that the type species of
A8 ... Vizzini & al.
Gymnopus (G. fusipes), Micromphale Gray (M. foetidum) and Setulipes Antonin (S. androsaceus) cluster within the same clade and that Marasmiellus Murrill represents an artificial, polyphyletic genus, suggesting that some Marasmiellus species should be included with Setulipes and Micromphale within Gymnopus.
Analyses of ITS and LSU rDNA sequences place our fungus in the Gymnopus dryophilus complex, a group of species belonging to section Levipedes (Fr.) Halling, subsection Levipedes (Antonin & Noordeloos 1997) (Tab. 1). This subsection traditionally encompasses species that share the following anatomical features: a pileipellis composed of relatively short, inflated, branched, loosely arranged hyphae, usually not coralloid or diverticulate (forming the so-called dryophila-structure, with elements which resemble a jig-saw puzzle, best seen in paradermal scalps); well-differentiated cheilocystidia, and pileipellis hyphae that do not turn green in alkali.
G. dryophilus could be conceived as a complex of morphologically similar species. This complex was clearly distinguished from the rest of the section Levipedes by both morphologically based phenetic and cladistic inferences (Vilgalys 1986), and molecular analyses (Wilson et al. 2004, Mata et al. 2006). The presence of mating intersterile groups within this species complex (“biological species”) from both North America (Vilgalys & Miller 1983) and Europe (Vilgalys & Miller 1987b) was also supported by DNA/DNA hybridization experiments (Vilgalys 1991). Concurrent morphological studies also demonstrated small morphological differences among species recognized within either continent (Vilgalys & Miller 1983, 1987a). These species and their infraspecific taxa could be distinguished based on a combination of shape and size of cheilocystidia together with rhizomorphs and gill colour.
Wilson et al. (2004) and Mata et al. (2006) are the first to attempt to circumscribe the taxa within this complex using ITS sequences. Both Bayesian and parsimony analyses of sequences within sect. Levipedes and the G. dryophilus complex show little variation and differ from one another by only 5- 10 bp (Mata et al. 2006). Morphological plasticity and lack of resolution of ITS sequence analyses complicate assignment of complex members to morphotaxa. Mata et al. (2006) observe that the G. dryophilus complex seemingly has a Euro- American distribution; this may be an artefact resulting from poor specimen sampling.
Within this section and complex, G. inexpectatus is well characterized by (1) a stipe with poorly developed basal mycelium, (2) growth on Quercus ilex wood debris, (3) a trichoderm-like pileipellis comprising coralloid digitate elements, irregular to subcoralloid cheilocystidia, and (4) broadly ellipsoid spores (Q = © 1.5). These results show that sect. Levipedes (Fr.) Halling s. Antonin et al. (1997) may also contain species with coralloid pileipellis cells as an extreme case of dryophila-structure.
Gymnopus inexpectatus sp. nov. (Italy) ... 49
Table 1. Similarities of G. inexpectatus ITS and LSU rDNA sequences to those retrieved from the GenBank database.!
ACCESSION # DESCRIPTIONS IDENTITY (%) E*VALUE ITS DQ480098 Gymnopus dryophilus voucher Duke 09 7e-114 DQ480097 Gymnopus dryophilus Duke 242 99 7e-114 DQ449960 Gymnopus ocior voucher TFB4284 2) 7e-114 DQ449959 Gymnopus ocior voucher TFB9015 99 7e-114 DQ449957 Gymnopus ocior voucher TFB3861 SPS) 7e-114 LOLA BE PE ah NE a ua al Ol nbd E@b waale AF042595 Collybia dryophila isolate RV83/180 99 0.0 AY640619 Gymnopus dryophilus isolate AFTOL-ID 559 99 0.0 AF291305 Gymnopus dryophilus 28S large subunit rDNA, 99 0.0 partial sequence AY639411 Gymnopus bicolor voucher AWW116-SFSU on 0.0 AF261328 Micromphale foetidum strain JEJ.VA.567 97 0.0
‘Search method: algorithm BLASTN 2.2.17 (Jun. 24, 2007) available at the NCBI site (http://www.ncbi.nlm.nih. gov/BLAST/), performed on Febr. 8, 2008.
(*) The expected E value represents the estimated number of sequences randomly corresponding (the lower the value, the better the correspondence with the sequence in the database).
None of the Gymnopus monographs from Europe (Antonin & Noordeloos 1997, 2008; Bon 1999), the Americas (Vilgalys & Miller 1983, Halling 1983, 1996, 1997; Mata et al. 2006), or Asia (Wilson et al. 2004) include a taxon close to the new species proposed here. The most closely related described species seems to be G. dryophilus var. lanipes (Malencon & Bertault) A. Ortega et al. (Malencon & Bertault 1975, Lonati 1986, Bon 1999, Vila & Llimona 2002, Ortega et al. 2003, Antonin & Noordeloos 2008), which exhibits similar habit and microscopical features. However, it differs from G. inexpectatus by having a non-insititious stipe, long stipe hairs, a more distinctly “jigsaw” (dryophila-like) pileipellis structure, and different spore size (E = 1.6-2.3, Q =1.85; Ortega et al. 2003). G. dryophilus var. lanipes typically grows in Mediterranean thermophilous forests (Morocco, Italy and Spain) among vegetal debris (such as Quercus ilex, Pinus spp., or, more rarely, Cistus spp). Ortega et al. (2003), who studied the holotype (n° 4508) in Malencon’s Montpellier herbarium (MPU), notes that the holotype material does not correspond to the taxon described in the protologue. It has a pubescent stipe, but its spores are larger (9-11 x 4-5 um), the pileipellis is a cutis of typical filamentous hyphae without a dryophila-structure, and the basidiomata are brown-reddish: this collection may represent a taxon within the G. terginus group. Because Malencon knew G. terginus very well, Ortega et al. (2003) supposed that a designation of the material as holotype was surely an unintentional error and selected a neotype from Iberian material that fitted the original description in all aspects, keeping the original name.
50 ... Vizzini & al.
One recently described Spanish species, G. bisporus (J. Carbo & Pérez-De-Greg.) J. Carbo & Pérez-De-Greg. [= G. catalonicus (Vila & Llimona) Vila & Llimona 2006, fide Antonin & Noordeloos 2008)], differs from G. inexpectatus in having clavate cheilocystidia, mono- and bisporic basidia and spores (8)8.9-10.6(12) x (3.5)4-4.9(5.5) um in size (Pérez-De-Gregorio & Carbé 2002, Antonin & Noordeloos 2008).
Based only on macro- and micromorphological features, G. inexpectatus could be mistaken for a Marasmiellus, as interpreted by Antonin & Noordeloos (1993), of sect. Dealbati Singer, subsect. Quercini Singer (with coloured pileus; Singer 1973, Desjardin 1997). In Europe (Antonin & Noordeloos 2008), the species belonging to the M. carneopallidus/M. mesosporus/M. maas-geesterani/ M. maritimus group may look like G. inexpectatus. M. carneopallidus (Pouzar) Singer, which has similar pileipellis structure and caulocystidia, can be distinguished by larger spores (7.5-12.5 x 4.5-6.5 um) and broadly clavate simple cheilocystidia. M. mesosporus Singer, with similar cheilo- and caulocystidia, has larger spores (10.5-14.5 x 5-7.5 um) and larger basidia (25-40 x 8-12 um) (Takehashi et al. 2007). M. maas-geesterani Robich & E. Campo has larger spores [(9)10-—12(13.5) x 5-6.5(7.5) um], differently shaped cheilo- and caulocystidia, and larger basidia (35-45 x 6.5-8.5 um) (Robich & Campo 2000). M. maritimus Contu & Noordel (Noordeloos & Antonin 2008 = Marasmiellus roseotinctus Contu & Noordel., nom. illeg., non Pegler) has larger spores (10-14.5 x 4-6 um) and larger basidia (22-35 x 7-10.5 um) (Noordeloos & Contu 2007).
In comparison with similar South-American species described by Singer (1973), M. xerophyticus Singer has a smaller, 7-13 mm broad, brown pileus with whitish margin, larger spores [(8)9-11 x (4.2)5-6(7) um], larger basidia (19-36 x 6-7.5 um), and differently shaped cheilocystidia; M. aurantiorufescens Singer has an ochraceous pileus that dries to rufous, longer and narrower spores [(4.5)5.5-6.7 x 2.5-2.8 um], differently shaped cheilocystidia, and growth on oak leaves; M. dryogeton Singer has a smaller (5-11 mm) ochraceous brown to cinnamon brown pileus, larger spores (6.2-8.5 x 3-3.5 um), smaller cheilocystidia (17-20 x 2-8 um), different caulocystidia, and grows on oak leaves; M. enodis Singer has a brown to deep brown, radially sulcate pileus, larger spores [6.5-9 x 2.5-4(4.5) um], differently shaped and smaller (22-35 x 7-9.5 um) cheilocystidia and caulocystidia in the form of Crinipellis-like hairs, and grows on dicotyledonous wood; M. dendroegrus Singer has an ochraceous- cinnamon, sulcate and striped pileus, larger spores (6-8.5 x 2.8-4.5 um), | differently shaped cheilocystidia and grows on dead wood of dicotyledons; M. quercinus Singer has a chestnut to deep cinnamon, small (6-10 mm) pileus, differently shaped, single, ventricose to fusoid cheilocystidia, different stipe covering, and grows on leaves (less frequently on small sticks and acorns) of
Gymnopus inexpectatus sp. nov. (Italy) ... 51
Quercus spp. The four last mentioned species are further distinguished by a distinctly dark brown incrusted hypodermium.
Acknowledgments
We would like to thank Antonio Ortega (Departamento de Botanica, Facultad de Ciencias, Universidad de Granada, Espafia) and Jordi Vila (Societat Catalana de Micologia - Facultat de Biologia Departament de Biologia Vegetal, Unitat de Botanica, Universitat de Barcelona, Espafia) for providing helpful suggestions. The research of the third author was supported by the Ministry of Culture of the Czech Republic (MK00009486201).
Literature cited
Antonin V, Halling RE, Noordeloos ME. 1997. Generic concepts within the groups of Marasmius and Collybia sensu lato. Mycotaxon 63: 359-368.
Antonin V, Noordeloos ME. 1993. A monograph of Marasmius, Collybia and related genera in Europe. Part. I. Marasmius, Setulipes, and Marasmiellus. Libri botanici, vol. 8. IHW Verlag, Eching (Germany).
Antonin V, Noordeloos ME. 1997. A monograph of Marasmius, Collybia and related genera in Europe. Part. II. Collybia, Gymnopus, Rhodocollybia, Crinipellis, Chaetocalathus, and additions to Marasmiellus. Libri botanici, vol. 17. IHW Verlag, Eching (Germany).
Antonin V, Noordeloos ME. 2008. Marasmioid and Collybioid Fungi in Europe. IHW Verlag, Eching (Germany), in press.
Bon M. 1999. Flore Mycologique d'Europe. 5. Les collybio-marasmioides et ressemblants. Doc. Mycol. Mém. hors série n. 5. Lille (France).
Desjardin D. 1997. A synopsis of Marasmiellus in the southern Appalachian Mountains. Mycotaxon 65: 237-261.
Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.
Halling RE. 1983. The genus Collybia (Agaricales) in the northeastern United States and adjacent Canada. Mycol. Mem. 8: 1-148.
Halling RE. 1996. Notes on Collybia V. Gymnopus Section Levipedes in Tropical South America, with comments on Collybia. Brittonia 48 (4): 487-494.
Halling RE. 1997. A revision of Collybia s.1. in the northeastern United States and adjacent Canada. http://www.nybg.org/bsci/res/col/colintro.html (Last update: 10/19/2004).
Holmgren PK, Holmgren NH. 1998. (continuously updated). Index Herbariorum: A global directory of public herbaria and associated staff. New York Botanical Garden's Virtual Herbarium. http:// sweetgum.nybg.org/ih/ (accessed 28 February 2008).
Kjoller R, Rosendahl S. 2000. Detection of arbuscular mycorrhizal fungi (Glomales) in roots by nested PCR (polymerase chain reaction) and SSCP (single stranded conformation polymorphism). Plant Soil 226: 189-196.
Lonati G. 1986. Altri due miceti Maroccani-Laziali. Marasmius dryophilus var. lanipes Malencon et Bertault, Hebeloma pallidum Malencon. Boll. Assoc. Micol. Ecol. Romana 5: 15-18.
Malencon G, Bertault R. 1975. Flore des Champignons supérieurs du Maroc, vol. II. Fac. Sci. Rabat (Maroc).
Mata JL, Hughes KW, Petersen RH. 2004. Phylogenetic placement of Marasmiellus juniperinus. Mycoscience 45: 214-221.
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Mata JL, Hughes KW, Petersen RH. 2006. An investigation of Omphalotaceae (Fungi, Euagarics) with emphasis on the genus Gymnopus. Sydowia 58: 191-289.
Noordeloos ME. 1995.-Genus Collybia. In: Bas C, Kuyper TW, Noordeloos ME, Vellinga EC (eds.), Flora Agaricina Neerlandica. A.A. Balkema, Rotterdam (The Netherlands), pp. 106-123.
Noordeloos ME, Antonin V. 2008. Contribution to a monograph of marasmioid and collybioid fungi in Europe. Czech Mycol. 60(1): 21-27.
Noordeloos ME, Contu M. 2007. On two remarkable Marasmiellus species from Sardinia, Italy. Oster. Z. Pilzk. 16: 181-186.
Ortega A, Antonin V, Esteve-Raventés FE. 2003. Three interesting thermophilic taxa of Gymnopus (Basidiomycetes, Tricholomataceae): G. pubipes sp. nov., G. pubipes var. pallidopileatus var. nov. and G. dryophilus var. lanipes comb. nov. Mycotaxon 85: 67-75.
Pérez-De-Gregorio MA, Carbé J. 2002. Una nueva especie de Collybia (Fr.: Fr.) Staude, eneontrad? en Catalufa. Revista Catalana Micol. 24: 277-282.
Robich G, Campo E. 2000. Marasmiellus maas-geesterani, una nuova specie dall ‘Italia. In: Micologia 2000. AMB, Fondazione Centro Studi Micologici, Trento (Italia), pp. 463-470.
Singer R. 1973. The genera Marasmiellus, Crepidotus and Simocybe in the Neotropics. Nova Hedwigia Beih. 44: 1-339.
Takehashi S, Kasuya T, Kakishima M. 2007. Marasmiellus mesosporus, a Marasmius-blight fungus newly recorded from sand dunes of the Japanese coast. Mycoscience 48: 407-410.
van Tuinen D, Zhao B, Gianinazzi-Pearson V. 1998. PCR in studies of AM fungi: from primers to application. In: Varma, A.K. (Ed.), Mycorrhiza Manual. Springer-Verlag, Heidelberg (Germany), pp. 387-399.
Vila J, Llimona X. 2002. Noves dades sobre el component flingic de les comunitats de Cistus de Catalunya. Revista Catalana Micol. 24: 75-121.
Vila J, Llimona X. 2006. Noves dades sobre el component flingic de les comunitats de Cistus de Catalunya. II. Revista Catalana Micol. 28: 167-207.
Vilgalys R. 1986. Phenetic and Cladistic Relationships in Collybia sect. Levipedes (Fungi: Basidiomycetes). Taxon 35 (2): 225-233.
Vilgalys R. 1991. Speciation and species concepts in the Collybia dryophila complex. Mycologia 83 (6): 758-773.
Vilgalys R, Miller OK Jr. 1983. Biological species in the Collybia dryophila Group in North America. Mycologia 75 (4): 707-722.
Vilgalys R, Miller OK Jr. 1987a. Morphological studies on the Collybia dryophila group in Europe. Trans. Brit. Mycol. Soc. 88: 461-472.
Vilgalys R, Miller OK Jr. 1987b. Mating relationships within the Collybia dryophila group in Europe. Trans. Br. Mycol. Soc. 89: 295-300. —
White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T (eds.), PCR Protocols. A Guide to Methods and Application. Academic Press, London (UK), pp. 315-322.
Wilson AW, Desjardin DE. 2005. Phylogenetic relationships in the gymnopoid and marasmioid fungi (Basidiomycetes, euagarics clade). Mycologia 97 (3): 667-679.
Wilson AW, Desjardin DE, Horak E. 2004. Agaricales of Indonesia. 5. The genus Gymnopus from Java and Bali. Sydowia 56: 137-210.
MYCOTAXON
Volume 105, pp. 53-58 July-September 2008
Phellinus mori sp. nov. (Basidiomycota, Hymenochaetales) from northern China
YU-CHENG Dar
*Corresponding author, yuchengd@yahoo.com Institute of Microbiology, Beijing Forestry University Beijing 100083, China
BaAo-Kat Cul
baokaicui@yahoo.com.cn Institute of Microbiology, Beijing Forestry University Beijing 100083, China
WAN-QIANG TAO
twq@bifb.gov.cn Station of Forest Protection, Beijing Municipal Bureau of Landscape & Forestry Beijing 100029, China
Abstract — A new resupinate member of the Hymenochaetaceae, Phellinus mori, is described from northern China. It belongs to the Phellinus laevigatus complex, but differs by strongly cracked basidiocarps when mature or dry, subparallel skeletals along the tubes, presence of cystidioles, ovoid or subglobose basidiospores, and habitat on Morus exclusively.
Key words — lignicolous fungi, poroid fungi, polypore, taxonomy
Introduction
During an investigation on lignicolous and poroid fungi in China, several specimens were collected on Morus from Heilongjiang Province and Beijing area in northern China. These collections are macroscopically similar to Fomitiporia punctata (P. Karst.) Murrill by resupinate and cushion-shaped basidiocarps, but they become strongly cracked when mature or dry. White mycelial strands are common in old tubes. Microscopically there are many hymenial setae and hyaline, fairly thick-walled, nondextrinoid and moderately cyanophilous basidiospores. The collections belong to the Phellinus laevigatus
54 ... Dai, Cui & Tao
complex but differ sufficiently from all Bevioush described taxa to be described as a new species.
Materials and methods
The studied specimens are deposited at the Herbarium of Beijing Forestry University (BJFC) and the Herbarium of Institute of Applied Ecology, Chinese Academy of Sciences (IFP). The microscopic procedure follows Dai (1999). Sections were studied at magnification up to x1000 by using a Nikon Eclipse E600 microscope and phase contrast illumination. Drawings were made with the aid of a drawing tube. In presenting the variation in the size of the spores, 5% of measurements were excluded from each end of the range, and are given in parentheses. In the text the following abbreviations are used: IKI = Melzer’s reagent, IKI- = negative in Melzer'’s reagent, KOH = 5% potassium hydroxide, CB = Cotton Blue, CB+ = cyanophilous, CB- = acyanophilous, L = mean spore length (arithmetic average of all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the L/W ratios between the specimens studied, n = number of spores measured from given number of specimens. Special colour terms followed Petersen (1996) and Anonymous (1969).
Description
Phellinus mori Y.C. Dai & B.K. Cui, sp. nov. Figs.1-3 MycoBank MB 511704)
Carpophorum perenne, resupinatum. Facies pororum cinnamomeum vel brunneum, pori rotundi, 7-8 per mm. Systema hypharum dimiticum, hyphae generatoriae septatae sine fibulis, hyphae skeletales contextum 2-4 um in diam. Setae sdsunt in hymenio, 11.6-26 x 5-8.4 um. Sporae ovoideae vel subglobosae, pallidae, crassitunicatae, 4.5-5.2 x 3.8-4.6 um.
Type. — China. Heilongjiang Prov., Ning’an County, Jingbohu Nature Reserve, on living tree of Morus, 8.1X.2007 Dai 8363 (holotype in IFP, isotypes in BJFC & H).
ETYMOLOGY — mori (Lat.): referring to the host tree genus.
Fruirnopy — Basidiocarps perennial, resupinate, firmly attached to the substrate, not readily separable, without odour or taste when fresh, woody hard and medium in weight when dry, up to 15 cm long and 6 cm wide, and 1 cm thick, usually receding and becoming cushion-shaped with age. Pore surface cinnamon brownish when fresh, becoming dark brown when bruised, clay buff and strongly cracked when mature or dry, with a slightly silky sheen; sterile margin narrow to almost lacking, concolorous with pores; pores mostly ~ circular, some slightly sinuous, (6—)7—8(-—10) per mm (n=90/3); dissepiments thin, entire. Context present at each layer, cinnamon brown to fawn brown, hard corky, ca. 0.1 mm thick. Tubes concolorous with pores, woody hard; white mycelial strands usually present in old tubes; tube layers distinct.
Phellinus morus sp. nov. (China) ... 55
Figs. 1-2. Phellinus mori. Fig. 1. Fresh basidiocarp. Fig. 2. Dried basidiocarp.
56 ... Dai, Cui & Tao
b-f:
10 pm
Fig. 3. Microscopic structures of Phellinus mori (drawn from the holotype). —a: Basidiospores. __b: Basidia and basidioles. —c: Cystidioles. —d: Setae. —e: Hyphae from trama. —f: Hyphae from
context.
Phellinus morus sp. nov. (China) ... 57
HyPHAL STRUCTURE — Hyphal system dimitic; all septa without clamp connections; tissue darkening but otherwise unchanged in KOH.
CONTEXT — Generative hyphae hyaline to pale yellowish, thin-walled, frequently branched and simple septate, 2-3.2 um in diam; skeletal hyphae dominant, rust brown, thick-walled with a narrow lumen to subsolid, unbranched, rarely septate, flexuous, interwoven, 2.5-4 um in diam; hyphae of white mycelial tufts, hyaline, thin-walled, strongly branched and flexuous, 1-1.8 um in diam.
TuBes — Generative hyphae hyaline, thin-walled, occasionally branched, frequently septate, 1.5-2.5 um in diam; skeletal hyphae dominant, rust brown, thick-walled with a narrow lumen to subsolid, more or less straight to flexuous, subparallel along the tubes, 2.2-3.5 um in diam. Setae frequent, ventricose to subulate, some furcate, dark brown, thick-walled, 11-24 x 5-8.5 um; fusoid cystidioles occasionally present, 8.7-14.7 x 3.4—5.2 um; basidia barrel-shaped, with four sterigmata and a simple septum at the base, 9-13.2 x 5.3-8 um; basidioles in shape similar to basidia, but slightly smaller; secondary hyphae of mycelial strands hyaline, thin-walled, frequently branched and septate, 1.5-2 ttm in diam. Rhomboid crystals frequently present in trama and hymenium.
SPORES — Basidiospores mostly ovoid, sometimes subglobose, hyaline, fairly thick-walled, smooth, IKI-, moderately CB+, (4—)4.3-5.2(-5.4) x (3.5-)3.8- 4.6(-4.8) um, L=4.74 um, W=4.16 um, Q=1.11-1.31 (n=210/7). ADDITIONAL SPECIMENS (PARATYPES) EXAMINED. — China. Beijing, Daxing County, Gusangyuan, dead branch of living Morus, 4.[X.2007 Dai 8291, 8292, 8293, 8294, 8295, 8297, 8298, 8300, 8301, 8303 & 8305 (IFP & BJFC). Heilongjiang Prov., Ning’an County, Jingbohu Nature Reserve, on living tree of Morus, 8.IX.2007 Dai 8309, 8310, 8339, 8342, 8344 & 8353 (IFP & BJFC).
Discussion
Phellinus mori belongs to the P. laevigatus complex, which includes P. betulinus (Murrill) Parmasto, P laevigatus (P. Karst.) Bourdot & Galzin, P prunicola (Murrill) Gilb., P rhamni (Bondartseva) H. Jahn, and P. spiculosus (W.A. Campb. & R.W. Davidson) Niemela. Parmasto (2007) studied the complex and provided statistical analyses of the basidiospores.
Phellinus mori resembles P. laevigatus s. str., but basidiospore shapes differ in the two species: ovoid or subglobose in P. mori, while ellipsoid in P laevigatus s. str. In addition, basidiospores of P. laevigatus s. str. (4.56-5.46 x 3.58-4.15 um, L=5.01 um, W= 3.87 um, Q=1.21-1.39, Parmasto 2007) are narrower than those in P mori. Macroscopically, the basidiocarp of Phellinus mori is strongly cracked when mature or dry, while it is not cracked in P. laevigatus s. str.
According to Parmasto (2007) Phellinus laevigatus occurs in Europe, West Siberia and Middle Asia, and its hosts are exclusively trees of Betula. Phellinus
58 ... Dai, Cui & Tao
mori was found in Beijing and Heilongjiang in northern China, growing exclusively on dead branches and living trees of Morus.
Phellinus mori is morphologically similar to P. betulinus, and the latter species occurs in China (treated as P. cf. laevigatus, Dai 1999). But P. betulinus has distinctly smaller basidiospores (3.25-4.55 x 2.51-3.49 um, Parmasto 2007), and it has been found so far only on Betula.
Among the members of the Phellinus laevigatus complex, basidiospores of P. prunicola, P. rhamniand P. spiculosus are 5.21-6.43 x 3.86—4.63 um, 5.46—-5.78 x 4.22-4.82 um and 4.99-6.1 x 3.83-5.23 um respectively (Parmasto 2007), and P. mori differs from these species by smaller basidiospores. Basidiospores of Phellinus spiculosus and P. mori may be slightly overlapped, but the former has basidiocarps not cracked and lacks cystidioles. Ecologically Phellinus spiculosus causes cankers on Quercus and Carya (Gilbertson and Ryvarden1987), but P. mori does not cause cankers, and lives on Morus exclusively.
Acknowledgements
We are grateful to Drs. Mario Rajchenberg (Centro Forestal CIEFAP, Argentina) and Zheng Wang (Yale University, USA) for reviewing the manuscript. The study is supported by the Department of Beijing Science and Technology (Project No. D0705002040391) and the Ministry of Science and Technology of China (Project No. 2005DFA30280).
Literature cited
Anonymous. 1969. Flora of British fungi. Colour identification chart. Her Majesty’s Stationery Office, London.
Dai YC. 1999. Phellinus sensu lato (Aphyllophorales, Hymenochaetaceae) in East Asia. Acta Bot. Fennica 166: 1-115.
Gilbertson RL, Ryvarden L. 1987. North American polypores 2. Fungiflora, Oslo. 452 pp.
Parmasto E. 2007. Phellinus laevigatus s.l. (Hymenochaetales): a ring species. Folia Cryptog. Estonica 43: 39-49.
Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til Svampekundskabens Fremme, Greve.
MYCOTAXON
Volume 105, pp. 59-64 July-September 2008
Perenniporia minor (Basidiomycota, Polyporales), a new polypore from China
HONG-XIA XIONG
xiongxiongbao@126.com Institute of Applied Ecology, Chinese Academy of Sciences Shenyang 110016, China
Graduate University, Chinese Academy of Sciences Beijing 100039, China
Yu-CHENG DAT
*Corresponding author, yuchengd@yahoo.com Institute of Applied Ecology, Chinese Academy of Sciences Shenyang 110016, China
BaAo-Kali CuI
baokaicui@yahoo.com.cn Institute of Microbiology, Beijing Forestry University Beijing 100083, China
Abstract — A new polypore, Perenniporia minor, collected from Changbaishan Nature Reserve, northeastern China, is described and illustrated. It is a saprophyte growing on fallen angiosperm branches. Macroscopically it differs from other species in the genus by tiny and pileate basidiocarps. Unlike most species of the genus, P minor is characterized microscopically by almost negative skeletal hyphae in Melzer’s reagent. P. minor and P. truncatospora have more or less similar basidiospores, but the latter has smaller pores, and its basidiospores are nondextrinoid.
Key words — lignicolous and poroid fungi, Polyporaceae, taxonomy
Introduction
Perenniporia Murrill is a cosmopolitan genus with around 90 species described and accepted (Gilbertson & Ryvarden 1987, Ryvarden & Gilbertson 1994, Hattori & Lee 1999, Decock & Figueroa 2000, Decock 2001a, b; Decock et al. 2000, 2001; Nufiez & Ryvarden 2001, Dai et al. 2002, Decock & Ryvarden1999a,
60 ... Xiong, Dai & Cui
b, 2000, 2003a, b). Twenty-three species in the genus have been recorded from China (Dai et al. 2002, Cui et al. 2006, 2007), and most of them occur in the tropics or subtropics. During an investigation on wood-inhabiting fungi in northeastern China, a tiny polypore was found that after microscopical examination turned out to represent an undescribed species of Perenniporia.
Materials and methods
The studied specimens are deposited at the Herbarium of Institute of Applied Ecology, Chinese Academy of Sciences (IFP). Anatomy was studied, and measurements and drawings were made from slide preparations stained with Cotton Blue. Drawings were made with the aid of a drawing tube. The microscopic routine used in the study followed Yuan et al. (2006). In presenting the variation in the size of the spores, the 5% of the measurements at each end of the range are shown in parentheses. The following abbreviations are used: IKI = Melzer’s reagent, IKI- = negative in Melzer’s reagent, KOH = 5% potassium hydroxide, CB = Cotton Blue, CB+ = cyanophilous, CB- = acyanophilous, L = mean spore length (arithmetic average of all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the L/W ratios between the specimens studied, n = number of spores measured from given number of specimens. Special colour terms are from Petersen (1996).
Description
Perenniporia minor Y.C. Dai & H.X. Xiong, sp. nov. Fig.1 MycosBank MB 511703
Carpophorum annuum, solitarium, pileatum; facies pororum cremea, pori rotundi, 4-6 per mm. Systema hypharum dimiticum vel trimiticum, hyphae generatoriae fibulatae, hyphae skeletales contextum 2.7-5 um in diam. Sporae pallidae, ellipsoideae vel truncatae, crassitunicatae, dextrinoidae, CB+, 4.9-6.2 x 3.8-4.5 um.
Type. — China. Jilin Prov., Antu County, Changbaishan Nature Reserve, Huangsongpu, alt. 1100 m, on fallen branch of Acer, 14.1X.2007 Dai 9198 (holotype in IFP, isotype in BJFC). i
ETyMOLOoGy — minor (L.), referring to the tiny basidiocarp.
FRUITBODY — Basidiocarps annual, pileate, solitary, soft corky, without odour or taste when fresh. Pileus semicircular to spathulate, projecting up to 0.8 cm, 1 cm wide, and 0.3 cm thick. Pileal surface cream to very pale buff when fresh, smooth, indistinctly concentrically zonate, becoming cinnamon-buff to pale brick, distinctly concentrically zonate or sulcate when dry. Pore surface cream when fresh, becoming cinnamon-buff when dry; pores round, 4-6 per mm, tube mouths fairly thick, entire. Context white when fresh, becoming cream and corky when dry, up to 0.1 cm thick; with a very thin cuticle present at the
Perenniporia minor sp. nov. (China) ... 61
Fig. 1. Microscopic structures of Perenniporia minor (drawn from the holotype). —a: Basidiospores. —b: Basidia and basidioles. —c: Hyphae from trama. —d: Hyphae from context.
62 ... Xiong, Dai & Cui
upper surface, cinnamon-buff. Tubes concolorous with pore surface, darker in contrast with context, hard corky, up to 0.2 cm long.
HYPHAL STRUCTURE — Hyphal system dimitic to trimitic; generative hyphae with clamp connections; skeletal hyphae IKI-, or very weakly amyloid, CB+; tissue unchanged in KOH.
CONTEXT — Generative hyphae infrequent, hyaline, thin-walled, occasionally branched, 2.2-3.5 um in diam.; skeletal hyphae dominant, hyaline, thick-walled, mostly with a narrow lumen, occasionally branched (trimitic-like), flexuous, interwoven, 2.7—5 um in diam.
TUBES — Generative hyphae infrequent, hyaline, thin-walled, occasionally branched, 2-3 um in diam.; skeletal hyphae dominant, hyaline, thick-walled with a narrow lumen, occasionally branched, flexuous, interwoven, 2.5-4.3 um in diam. Hymenium usually collapsed, basidia usually infrequent, barrel- shaped to pyriform, with four sterigmata and a basal clamp connection, 13- 16 x 4.5-6.5 um; basidioles in shape similar to basidia, but slightly smaller. Rhomboid crystals frequently present in trama and hymenium.
SpoRES — Basidiospores ellipsoid, mostly truncate, hyaline, thick-walled,
smooth, usually bearing a small guttule, dextrinoid, CB+, (4.7—)4.9-6.2(-7) x
(3.6-)3.8-4.5(-4.7) um, L=5.36 um, W=4.12 um, Q=1.26-1.35 (n=96/3). ADDITIONAL SPECIMENS (PARATYPES) EXAMINED. — China. Jilin Prov., Antu County, Changbaishan Nature Reserve, Huangsongpu, alt. 1100 m, on fallen angiosperm branch, 29.X1.2007 Wei 3500 (IFP). Fusong Country, Lushuihe, Hongwei, on fallen angiosperm branch, 28.X1.2007 Wei 3467 (IFP).
REMARKS — In the field Perenniporia minor resembles a juvenile Tyromyces
chioneus (Fr.) P. Karst., but the latter is readily differentiated microscopically by
its thin-walled, nondextrinoid basidiospores.
The new species differs from other taxa in the genus by its tiny and pileate basidiocarp (less than 1 cm), and almost negative skeletal hyphae in Melzer’s reagent. Perenniporia contraria (Berk. & M.A. Curtis) Ryvarden, P truncatospora (Lloyd) Ryvarden, P ohiensis (Berk.) Ryvarden and P. ochroleuca (Berk.) Ryvarden also have small basidiocarps when young, but these species are perennial. In addition, P contraria has smaller basidiospores (3.4-4 x 2.5-3.1 um, Cui et al. 2006), and spores of P. ohiensis (12-14 x 7-8.8 um, Dai et al. 2002) and P. ochroleuca (9-12 x 5.5-7.5 um, Dai et al. 2002) are distinctly bigger than those of P minor. P. truncatospora resembles P. minor in basidiospore shape and size (5.3-7.5 x 4-5 um vs. 4.9-6.2 x 3.8-4.5 tm), but the former has smaller pores (6-8 per mm) and basidiospores that are negative in Melzer’s reagent.
Unlike most species of Perenniporia, skeletal hyphae of P minor are nondextrinoid, but this character is not unique to the species. In fact, several species of Perenniporia have non- to faintly dextrinoid skeletal hyphae, e.g.,
Perenniporia minor sp. nov. (China) ... 63
P. medulla-panis (Jacq.) Donk and P. ohiensis; however, basidiospores of these species are dextrinoid and cyanophilous. We consider the spore Melzer’s and Cotton Blue reactions as more important and consider them as key characters for the genus.
Acknowledgements
We are grateful to Drs. Mario Rajchenberg (Centro Forestal CIEFAP, Argentina) and Zheng Wang (Yale University, USA) for reviewing the manuscript. The study is supported by the National Natural Science Foundation of China (Project No. 30771730) and the Ministry of Science and Technology of China for research grant (Project No. 2005DFA30280).
Literature cited
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Cui BK, Dai YC, Decock C. 2007. A new species of Perenniporia (Basidiomycota, Aphyllophorales) from China. Mycotaxon 99: 175-180.
Dai YC, Niemela T, Kinnunen J. 2002. The polypore genera Abundisporus and Perenniporia (Basidiomycota) in China, with notes on Haploporus. Ann. Bot. Fennici 39: 169-182.
Decock C. 2001a. Studies in Perenniporia. Some Southeast Asian taxa revisited. Mycologia 93: 774-795.
Decock C. 2001b. Studies in Perenniporia (Basidiomycetes, Polyporales): African taxa 1. Perenniporia dendrohyphidia and Perenniporia subdendrohyphidia. Syst. Geogr. PJ. 71: 45-51.
Decock C, Buchanan P, Ryvarden L. 2000: Revision of some Australasian taxa of Perenniporia (Basidiomycetes, Aphyllophorales). Australian Syst. Bot. 13: 823-844.
Decock C, Figueroa H. 2000. Studies in Perenniporia, Navisporus ortizii,a synonym of Perenniporia martius, and a note on Navisporus and Perenniporia in Cuba. Crypt. Mycol. 21: 153-162.
Decock C, Figueroa H, Ryvarden L. 2001. Studies in Perenniporia. Perenniporia contraria and its presumed taxonomic synonym Fomes subannosus. Mycologia 93: 196-204.
Decock C, Ryvarden L. 1999a. Studies in neotropical polypores. Some coloured resupinate Perenniporia species. Mycol. Res. 103: 1138-1144.
Decock C, Ryvarden L. 1999b. Studies in Perenniporia: Perenniporia detrita and its taxonomic synonyms. Mycologia 91: 386-395.
Decock C, Ryvarden L. 2000. Studies in neotropical polypores 6. New resupinate Perenniporia species with small pores and small basidiospores. Mycologia 92: 354-360.
Decock C, Ryvarden L. 2003a. Perenniporiella gen. nov. segregated from Perenniporia, including a key to neotropical Perenniporia species with pileate basidiomes. Mycol. Res. 107: 93-103.
Decock C, Ryvarden L. 2003b. Studies in Perenniporia (Basidiomycetes, Perenniporiales) Pseudopiptoporus chocolates comb. nov., a synonym of Perenniporia ahmadii, and a note on Pseudopiptoporus. Nova Hedwigia 77: 199-211.
Gilbertson RL, Ryvarden L. 1987. North American polypores 2. Megasporoporia - Wrightoporia. Fungiflora, Oslo. 434-885 pp.
Hattori T, Lee S. 1999. Two new species of Perenniporia described from a lowland rainforest of Malaysia. Mycologia 91: 525-531.
64 ... Xiong, Dai & Cui
Nufiez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Synopsis Fungorum 14: 165-522.
Petersen JH. 1996. Farvekort. The Danish Mycological Society's colour-chart. Foreningen til Svampekundskabens Fremme, Greve.
Ryvarden L, Gilbertson RL. 1994. European polypores 2. Synopsis Fungorum 7: 394-743.
Yuan HS, Li J, Huang MY, Dai YC. 2006. Antrodiella stipitata sp. nov. from Heilongjiang Province, northeastern China, and a critical checklist of polypores from the area. Cryptogamie Mycologie 272129,
Volume 105, pp. 65-78 July-September 2008
A taxonomic study of Heterodermia (Lecanorales, Ascomycota) in South Korea based on phenotypic and phylogenetic analysis
XIN Li WEI’”*, HENG Luo?*, YOUNG JIN KOH? & JAE SEOUN HuR”
*ishur1@sunchon.ac.kr ‘Key Laboratory of Systematic Mycology & Lichenology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China *Korean Lichen Research Institute, Sunchon National University Sunchon 540-742, Korea
Abstract — A study on taxonomy and phylogeny of Heterodermia in South Korea is presented for the first time. Phenotypic analysis was based on morphological, anatomical, and chemical characters, and phylogenetic analysis was based on nrDNA ITS sequences. The results suggest that the genus Heterodermia is monophyletic. Some characters (e.g. corticate vs. ecorticate lower surface, absence vs. presence of yellow pigments or soredia) are shown to be taxonomically significant within the genus. Of the eighteen species of Heterodermia known to occur in South Korea, eleven were confirmed in this study. Brief species diagnoses accompany a key to Heterodermia in South Korea, and comparisons with similar taxa are included.
Key words — lichen flora, macrolichen, Physciaceae, comprehensive analysis
Introduction
The genus Heterodermia belongs to the lichenized ascomycetous family Physciaceae Zahlbr. Heterodermia is mainly characterized by linear lobes and by spores of the Pachysporaria- or the Polyblastidia-type (Poelt 1965, Brodo et al. 2001). All species of Heterodermia were included in Anaptychia Korb. (Kurokawa 1962) until thick-walled spores and the presence of atranorin came to be regarded as useful characters for separating these genera (Poelt 1965, Swinscow & Krog 1976). Despite the many papers on Heterodermia (Awasthi 1973, Swinscow & Krog 1976, Wei 1991, Kurokawa 1998, Moberg & Nash 1999, Grube & Arup 2001, Nordin & Mattsson 2001, Kurokawa 1962), almost no studies on the genus were available for South Korea until the recent macrolichen flora by Park (1990) that treated 12 Heterodermia species and provided brief
*Xin Li Wei and Heng Luo contributed equally to this work
66 ... Wei & al.
Table 1. Seventeen Heterodermia specimens used for DNA extraction
CoLL.# SPECIES NAME LOCALITY, GPS, ALT. (m) ACCESSION # 040340 H. boryi Mt. Chiri, N35°19’07.0” E127°40°42.7°, 1580 EU045422 040754 H. boryi Jeju.Do, N33 21 32,9" 8126 31 54.97 1920 EU045423 050093 H. diademata Mt. Deogyu, N35°51’24.1” E127°44’53.67,1580 EU045425 050273 H. diademata Mt. Sorak, N38°09718.1” E128°19°46.1, 880 EU045426 060639 H. dissecta Mt. Chiri, N35°19'01.0” E127°44’30.1", 1160 EU045424 040658 H. hypochraea Jeju Do, N33°24’26.6” E126°29°46.0", 645 EU045427 060232 H. hypoleuca Mt. .Chir,,N3517935.3° B127°32750.3,1390 EU045428 050017 ~—«-H. hypoleuca Mt. Naejang, N35°29°46.1” E126°53’56.7”, 600 EU045429 050317 H. japonica Mt. Sorak, N38°09711.2” E128°26'24.2”, 1265 EU045431 060291 H. japonica Mt. Chiri, N35°18'14” E127°34’10.4”, 1430 EU045432 060173 H. microphylla Mt. Chiri, N35°17'25.78” E127°31’59.7°, 1420 EU045433 060494 H. microphylla Mt.Deogyo, N35°51°10.0” E127°44°56.9, 1568 EU045434 050136 H. obscurata Wando, N34°21'10.3” E126°41°10.9", 535 EU045435 040623 H. obscurata Jeju Do, N33°28'25.5” E126°29°47.7°, 145 EU045436 050493 H. pseudospeciosa Mt. Haegsok, N34°41’21.1” E126°40°47.5” EU045438 040619 H. subascendens Jeju Do, 145, N33°28'25.5” E126°29°47.7” EU045441 040847 H, subascendens Jeju Do, 370, N33°27'15.4” E126°33’41.1” EU045442
descriptions of each species and a key to the taxa. However, some ambiguous morphological characters left some taxonomic problems in need of additional research. The most recently published checklist of Korean lichens (Hur et al. 2005), reports twenty Heterodermia species from the Korean peninsula. Thus, in the present study, we have tried to clarify the taxonomy of Heterodermia in South Korea based on specimens collected during recent nationwide field surveys. In addition, we also evaluate whether Heterodermia is monophyletic and discuss the importance of certain morphological and chemical characters in differentiating species.
Materials and methods
MATERIALS — Two hundred and fifty-six specimens from South Korea have been examined and are deposited in the herbarium of the Korean Lichen Research Institute (KoLRI) in Sunchon, Korea. No type specimens were included in this study.
MorrHo.ocy — ‘The description of the external morphology is based on air-dried material observed under a dissecting microscope. For anatomical descriptions, sections were made with a razor blade under the stereomicroscope, mounted in GAW (glycerol: alcohol: water = 1:1:1), and observed under a light microscope (Olympus BX 50). Illustrations were taken with a digital camera (Nikon Coolpix 4500).
CHEMISTRY — Lichen substances were detected by the color reaction of a spot test (with KOH) and by thin-layer chromatography (with B solvent system) (Culberson 1972, White & James 1985).
Heterodermia in South Korea ... 67
Table 2. Sixty-nine phenotypic characters chosen for phenotypic analysis of
Heterodermia No. CHARACTER No. CHARACTER 1 Thallus foliose 36 Yellow pigment present in the lower surface 2 Thallus subfiliform oe Rhizines marginal 3 Lobes linear 38 Rhizines at lower surface 4 Lobes subdichotomously branched 39 Rhizines marginal and at lower surface >» Lobes irregularly branched 40 Rhizines black 6 Lobes discrete 4} Rhizines tan 7 Lobes crowded 42 Rhizines white 8 Lobe tips plane 43 Rhizines concolorous to thallus 9 Lobe tips ascending 44 Rhizines simple to squarrosely branched 10 Pape mnscitcinle 4s Paice dibs to subdichotomously 11 Lobe tips with pruina 46 Medulla white 12 The pruina of lobe tips round-patch 47 Pycnidia and (or) apothecia not seen 13 sti ie preps peace Get nlte 48 Pycnidia laminal, limited near tips 14 Lobe tips sometimes orange 49 Conidia bacilliform 15 Soredia present 50 Conidia small 16 Soredia marginal ba Conidia big 17 Soredia laminal Lys Apothecia lecanorine 18 Soredia marginal and laminal 53 Apothecia laminal 19 Soredia at lower surface 54 Apothecia terminal 20 Round-patch soralia 55 Apothecia marginal 21 Labriform soralia 56 Apothecia margin with lobules 22 Capitate soralia 57 The lobules of apothecia margin pruinose 23 Isidia present 58 Disks pruinose 24 Lobules present aS) Eight ascospores per asci 25 Lobules marginal 60 Ascospores brown 26 Lobules marginal and laminal 61 Ascospores 1-septate 27 Lobules tips suberect 62 Upper cortex prosoplectenchymatous 28 Lobules tips with pruina 63 Grayish surface layer present og Lobules isidioid 64 Gonidia green 30 Upper surface maculate 65 Lower cortex prosoplectenchymatous 31 Upper surface papillate 66 No substances contained in the thallus 32 Lower surface corticate 67 Atranorin and zeorin present 33 Lower surface white 68 Norstictic acid present 34 Lower surface pale brown 69 Norstictic and salazinic acids present 35 Lower surface arachnoid
68 ... Wei & al.
PHENOTYPIC ANALYSIS — The phenotypic analysis is based on sixty-nine phenotypic characters including morphological, anatomical, and chemical characters (Table 2). Maximum parsimony analysis was performed using PAUP version 4.0b10 (Swofford, 2002). The reliability of the inferred tree was tested by 1000 bootstrap replications. Anaptychia palmatula (Michx.) Vain. was used as outgroup for the analysis.
DNA EXTRACTION AND NRDNA AMPLIFICATION — Seventeen lichen thalli (Tablel) were fractioned with cryo-tissue-crasher (SK200, Tokken, Japan). Total DNA was extracted directly from whole thalli according to Ekman (1999) with DNeasy Plant Mini Kit (QIAGEN, Germany), then purified by PCRquick-spin™ PCR Product Purification Kit (iNtRON Biotechnology, INC.). ThenrDNAITS region (ITS1-5.8S-ITS2) was amplified by PCR. The primers for amplification were: ITS1F (5'-CTTGGTCATTTACAGGAAGTAA- 3'; Gardes & Bruns, 1993) and ITS4A (5°-ATTTGAGCTCTTCCCGCTTCA-3;; White el al. 1990). Conditions for PCR amplification and cycle sequencing have been described previously (Arup 2002).
SEQUENCING AND PHYLOGENETIC ANALYSIS — PCR products were sequenced using ABI 3700 automated DNA Sequencer in NICEM at Seoul National University. The phylogenetic analysis was executed with software Mega3.1 (Kumar et al., 2004). The Kimura 2-parameter model was selected, and gaps were retained initially while being excluded in the pairwise distance estimation. The neighbor joining (NJ) (Saitou & Nei 1987) method was used to construct the phylogenetic tree, reliability of the inferred tree was tested by 1000 bootstrap replications. Some Anaptychia species were available as outgroups for the phylogenetic analysis.
Results and discussion
Phenotypic analysis
Phenotypic analysis (Fig.1) showed that all treated taxa of Heterodermia form a monophyletic group, obviously separated from the outgroup Anaptychia palmatula with 98% bootstrap support. Within Heterodermia, two sections (I and II) based on the presence or absence of a lower cortex were indicated, suggesting that the character-pair ‘lower surface corticate or ecorticate’ is important to distinguish species.
Section I includes two species clusters (I-1 and I-2) according to the presence of soredia and lobules, respectively. These characters are crucial in the taxonomy among species with a lower cortex. In section IJ, no striking characters were found to correspond to the topology structure except the spot test character of the lower surface (K+, due to the presence of a yellow pigment). Three species with this character — H. obscurata, H. hypochraea, H. subascendens — and one species that sometimes expresses the character — H. japonica — clustered in II-1 to some extent, suggesting that the spot test reaction has taxonomic significance among species lacking a lower cortex. No other phenotypical characters were shown to have taxonomic significance in Heterodermia.
Heterodermia in South Korea ... 69
Heterodermia japonica 53
Heterodermia obscurata
Heterodermia hypochraea Hi-1 Lower surface Kt; 51 yellow pigment present
Heterodermia subascendens Il Lower surface ecorticate Heterodermia boryi
Heterodermia hypoleuca
Heterodermia microphylla
98
98
Heterodermia diademata |e Lobules present
Heterodermia dissecta
: : I Lower surface corticate 98 Heterodermia pseudospeciosa
: |u Soredia present Heterodermia speciosa
Anaptychia palmatula
Fig. 1. Maximum parsimony tree based on morphological, anatomical and chemical characters of 11 species of Heterodermia in Korea (Anaptychia palmatula = outgroup). Data matrix includes 12 taxa, 69 characters. All characters are of type ‘unord, all characters have equal weight, among which 13 characters are constant, 14 variable characters are parsimony-uninformative, number of parsimony-informative characters = 42. Tree length = 138, consistency index (CI) = 0.4058, homoplasy index (HI) = 0.5942, CI excluding uninformative characters = 0.3387, HI excluding uninformative characters = 0.6613, retention index (RI) = 0.4058, rescaled consistency index (RC) = 0.1647. Heuristic search, bootstrap=1000. The numbers in each node represents bootstrap support value, and the numbers lower than 50 were not shown.
Phylogenetic analysis
The NJ consensus tree (Fig.2) clusters all Heterodermia species into a well supported (100% support) monophyletic clade that is clearly separated from Anaptychia species. Two groups are included within the Heterodermia clade, with group I species corticate and group II species ecorticate. This result agrees with those from the phenotypic analysis and confirms that the presence or absence of a cortical layer on the lower surface is an important diagnostic character.
Within the group I, H. diademata, H. pseudospeciosa and H. dissecta are clearly separated. On the one hand, H. diademata and H. dissecta differ from H. pseudospeciosa by the absence or presence of soredia; on the other, H. diademata differs from H. dissecta by absence or presence of norstictic and salazinic acids, which are also present in H. pseudospeciosa. ‘This suggests that absence or presence of soredia or norstictic and salazinic acids are important in separating species in Group I.
70 ... Wei & al.
p A Heterodermia diademata EU045426 1001 A Heterodermia pseudospeciosa £U045438 A Heterodermia dissecta FU045424 , & Heterodermia hypoleuca EU045429 ; A Heterodermia hypoleuca EU045428 100 | A Heterodermia microphylla EU045433 61> & Heterodermia microphylla EU045434 A Heterodermia obscurata EU045436 199! & Heterodermia obscurata EU045435 Group if lower cortex absent 100, A Heterodermia subascendens EU045441 1007! A& Hetorodermia subascendens EU045442
~
| Group I: lower cortex present
100, A Heterodermia diademata FU045425 |]
1
|
98 |
Monophyletic clade
~ & Heterodermia hypochraea £U045427 | 100,;-— A Heterodermia boryi FU045422
63] 1A Heterodermia boryi EU045423 | A Heterodermia japonica EU045431 | 100! & Heterodenmia japonica EU045432 | Anaptychia palmatala DQ394376
Anaptychia ciliaris AF389939
Anaptychia runcinata AF 224364
une Anaptychia bryorum AF2507&1
Anaptychia ulotricoides AF 540494
— Anaptychia elburzuana AY368 145
76
100
an | 0.05
Fig. 2. NJ consensus tree based on nrDNA ITS sequences. Nucleotide: Kimura 2-parameter, pairwise deletion, bootstrap=1000. The numbers in each node represents bootstrap support value, and the numbers lower than 50 were not shown. The sequences marked with Awere obtained in this study, and others were downloaded from GenBank.
Group II shows species witha yellow pigment at the lower surface (H. hypochraea, H. subascendens, H. obscurata) in one cluster separated from species lacking this yellow pigment (H. boryi) or only sometimes possessing the pigment (H. japonica), suggesting that presence or absence of yellow pigment may serve as a helpful taxonomic character. Although there is only a short genetic distance separating H. hypoleuca and H. microphylla, we think they should be treated as two different species based on the phenotype showing two taxa are easily distinguished phenotypically by rhizine color and the presence or absence of soredia.
Based on our experimental results, we conclude that the genus Heterodermia forms a well-supported monophyletic clade. We also identify four characters as important for separating taxa within Heterodermia: (i) cortication of the lower surface (Fig.3A, B), (ii) presence or absence of soredia (in some cases, Fig.3D), (iii) presence or absence of a yellow pigment (Fig.3C), and (iv) the presence or absence of norstictic and salazinic acids.
Taxonomy
Based on the comprehensive analysis given above, a key to the species is presented, in which seven additional species not examined in our study but reported before in South Korea are also included. All treated taxa are subsequently listed and include diagnostic characters of each species. In case
Heterodermia in South Korea... 71
of large numbers of studied specimens only some representative specimens are cited.
Key to the species of Heterodermia in South Korea
i ORS CATE ay <2) AS a a oh, Se Sr) A eee H. isidiophora APASIC IA ADSCN Utes py hey ae ede etere pace es he ME an ce 2. 2, LOWenSUrlacercOrticatesss oa ia: jee aL NRC ACER aut Lae deca tt eee eee 3 Oe Lower surface noGeorticaté ainiiewe Ginn eRe eee eel tis ee ee eee a mi ZINES Inapeitial Neate". eeeeaneree omer tt ear, sone cP: eee H. rubescens Da IZINES NOM ar Cll alate cone ie ete. cyte eee es, oa eee cee nL ee er ae 4 A MLODESSOLCCIALC HORNS a) ie ates See Otis <p Caen ak tS | a ee D A MICODES COOLCCIALE Lo atts Noligtt avis aucde.t eae meres Sams ao ek tol Ea tee aed ae ee 6 PR NOFStICLIGIACICH present <4iiut.. Mei nee rene nea ee H. pseudospeciosa Da Norsticucacicabsent@. . aeiniee ys. 5 stip meet eanel ne cumin Proves tan, ete pe H. speciosa 6. Norstictic and salazinic acids absent; lobules marginal and laminal H. diademata 6. Norstictic and salazinic acids present; lobules only marginal............ H. dissecta 72 Lone andew nite, ciliapresentim: eft iraqake. Mat eee Genny sere eae ere H. comosa Failte absentee. hei ae aa.” pies ban ad tee. lvo alse. 0c eae 8 SMBVEC CELL ARV CLO Wieeemites caCuis Naret een Nagle Wee rem © or leet kt ef ene H. firmula SmIVICdULaWHItCte ee ee ee Cee eee ee 9 9. Lower surface K+ purple-red, yellow pigment present ......................-5. 10 DLowetsuttace Kesyellow pigmentabsent-a jacessiriacts ieee Seaiaases 15 TOMES esGrediaterts, 28102. el SMoiTi02 © Dit: .“olUGOl | LeOOoRL See noe lish AVE ARCRAU Rel fota TEC Le tora Plas Lh ek AUREL) Pot Mn A robataetn Rio hed ah iy oo 9 oe 14 itt SiLowerrstirtace purplesblagk avcenters A0s ci. Ars Hee cite tat akira See Deen le ivie WowelssUllace WhitesalCEN(Ch cage coc ts pineal concn Beene cae 13 12 URhizinestwiiteeeee weit ts ce er cae ose ot ROR Te ee aaa H. loriformis PORE IZiN eS DAC Kee terme ie tame e. cos ne So skac car cee eee H. dendritica 13s Unknown yellowsupstalice present. a5. 920i) Ot, Ge eine 4 aa, H. pandurata 15, Unknownryellow substancembsent: sacs = ta. Oe tam See eee H. hypochraea {AsSonediamtlower suriace non-labruormie orc ae ee H. subascendens 14 Leabriform. sora lian tie acc ste kat bekres cnet oby das ture cork ees eee H. obscurata 15: Thallus esorediatetn Wweahaeb es Face. igh ete seater eee H. hypoleuca [5othallistsoretiate wt oe ee ee ec ee cs ete + COR SUE Ere ie See ave. 16 VOusGkecialaininalemixed WithslODUleSs ges. .e ee nn eer eee H. microphylla LonSorediaimarginakorsséerminal 12.065 vive tlt yee eee nae ee 17 Wesercciaterminal thallus foliose. .utisn> sie ae cla ee eee eee eee H. japonica ivPsoredia marpinalsthallus'subfiliform yee Oe a ee get H. boryi
72. ... Wei & al.
Heterodermia Trevis., Atti Soc. Ital. Sci. Nat. (Milano) 11: 613. 1868 Lectotype: H. speciosa (Wulfen) Trevis. (selected by Poelt 1965).
1. Heterodermia boryi (Fée) K.S. Singh & S.R. Singh, Geophytology 6 (1): 33, 1976
REMARKS: H. boryi is easily characterized by its strap-shaped to subfiliform lobes, circinate tips, and the round or patch-shaped soralia that are lateral and laminal. SPECIMENS EXAMINED: KOREA. Mt. Jiri, N35°19°09.1”, E127°40726.9”, alt. 1620 (1564) m, on rock, 23 April 2004, Hur 040329; Mt. Jiri, N35°19°07.0” E127°40°42.7°, alt. 1580(1531) m, on rock, 23 April 2004, Hur 040340; Jejudo, N33°21°32.9" E126°31°54.9", alt. 1920 m, on rock, 27 August 2004, Hur 040754.
2. Heterodermia diademata (Taylor) D.D. Awasthi, Geophytology 3: 113, 1973
REMARKS: H. diademata is characterized by a sublinear thallus, the abundance of marginal and laminal lobules, and a corticate lower surface (Fig. 3B). The species is similar to H. dissecta in morphology, but differs in chemistry, salazinic and norstictic acids absent in H. diademata, while present in H. dissecta.
SELECTED (OF 30) SPECIMENS EXAMINED: KOREA. Mt. Naejang, alt. 680 m, on rock, 29
June 2003, Hur 030429; Baekdam temple, N38°11°16.4” E128°21’42.7°, alt. 450 m, on
rock, 11 October 2004, Hur 041509; Mt. Jiri, N35°20731.5” E127°41°08.9”, alt. 715 m, on
bark, 4 September 2004, Hur 040928; Mt. Deogyu, N35°51'24.1” E127°44’53.6", alt. 1580
m, on bark, 3 April 2005, Hur 050093; Mt. Sorak, N38°09'18.1” E128°19°46.1°, alt. 880 m,
on bark, 16 June 2005, Hur 050273; Wando arboretum, N34°21710.3” E126°41°10.9”alt.
535 m, on rock, 13 April 2005, Hur 050133.
3. Heterodermia dissecta (Kurok.) D.D. Awasthi, Geophytology 3: 113, 1973
REMARKS: H. dissecta is characterized by sublinear lobes, the abundance of marginal (occasionally laminal) lobules, and a corticate lower surface. The species is similar to H. diademata in morphology, but differs in chemistry, with salazinic and norstictic acids present in H. dissecta and absent in H. diademata.
SPECIMENS EXAMINED: KOREA. Mt. Baega, N34° 10°14’, E127°8°45’, alt. 500 m, on rock,
8 October 2005, Hur 050572. Mt. Chiri, N35°19°1”, E127°44’30.1", alt. 1160 m, on rock,
15 September 2006, Hur 060639.
4. Heterodermia hypochraea (Vain.) Swinscow & Krog, Lichenologist 8 (2): 119, 1976
REMARKS: H. hypochraea has a papillate upper surface, erect lobe tips, and an ecorticate
lower surface with a yellow pigment (Fig. 3C). The species is similar to H. hypoleuca in
morphology, but differs in its white rhizines, erect lobe tips, and yellow lower surface.
SPECIMEN EXAMINED: KOREA. Jejudo, N33°24'26.6” E126°29°46.0’, alt. 645 m, on bark, 26 August 2004, Hur 040658.
5. Heterodermia hypoleuca (Mihl.) Trevis., Atti Soc. ital. Sci. nat. (Milano) 11: 615, 1868
Remarks: H. hypoleuca has a maculate upper surface, an ecorticate lower surface (Fig. 3A), center that is reticulately veined, and hyaline to tan rhizines. This species
Heterodermia in South Korea... 73
Fig. 3. A. Ecorticated lower surface of Heterodermia hypoleuca, Hur 030174; B. Corticate lower surface of H. diademata, Hur 030429; C. Yellow pigment (arrow) at the lower surface of H. hypochraea, Hur 040658; D. Labriform soralia of H. pseudospeciosa, Hur 030204.
is similar to H. hypochraea, but differs by the tan colored rhizines, the plane lobe tips, and the absence of a yellow pigment in the lower surface. H. hypoleuca is also similar to H. japonica in morphology, but differs in that H. hypoleuca has hyaline to tan rhizines and lacks soredia.
SELECTED (OF 107) SPECIMENS EXAMINED: KOREA. Mt. Baekwoon, alt. 1050 m, on rock, 5 May 2003, Hur 030174; Mt. Hanla, alt. 1300, on moss, 22 June 2003, Hur 030333; Mt. Sobaek, N36°56’50.5” E128°29°51.2”, alt. 1110 m, 2 October 2003, Hur 030752; Mt. Gaya, N35°48’28.5” E128°07°00.6", alt. 965(916) m, on rock, 15 April 2004, Hur 040196; Mt. Odea, N37°43’42.8” E128°35736.6’, alt. 650(662) m, on Abies, 7 May 2004, Hur 040400; Jejudo, N33°25’21.9” E126°33’35.7°, alt. 505(614)m, on bark, 26 August 2004, Hur 040676; Mt. Taebaek, N37°06700.3” E128°57702.5”, alt. 1220 m, on bark, 12 September 2004, Hur 041046; Mt. Naejang, N35°29°46.1” E126°53°56.7’, alt. 600 m, on bark, 8 January 2005, Hur 050017; Mt. Deogyu, N35 51-292” E127 4503.4, alt 1530 m, on bark, 2 April 2005, Hur 050079; Mt.Chiri, N35°17°35.3” E127°32’50.3”, alt. 1390 m, on Quercus, 17 June 2006, Hur 20060232.
6. Heterodermia japonica (M. Sat6) Swinscow & Krog, Lichenologist 8 (2): 122, 1976
REMARKS: H. japonica is characterized by ascending lobe tips, labriform to subcapitate
soralia, an ecorticate lower surface, and the presence (‘H. propagulifera’) or absence
7A ... Wei & al.
(H. japonica s.str.) of a yellow pigment. This species is similar to H. hypoleuca, but differs in having black rhizines and labriform to subcapitate soralia at the lobe tips.
SELECTED (OF 46) SPECIMENS EXAMINED: KOREA. Mt. Baekwoon, alt. 705 m, on bark, 1 June 2003, Hur 030282; Mt. Taebaek, N37°06’21.7” E128°57'12.1yalt. 1025 m, on rock, 2 November 2003, Hur 030841;Jejudo,33°24’26.6” E126°29°46.0", alt. 645 m, on bark, 26 August 2004, Hur 040654; Mt. Sorak, N38°06'40.4” E128°24’26.8’, alt. 1335 m, on Abies, 10 October 2004, Hur 041460; Mt. Deogyu, N35°47°19.4” E127°42°30.3°, alt. 1269 m, on rock, Hur 050207; Mt.Chiri, N35°18'14” E127°34710.4”, on 1430 m, on rock, 17 June 2006, Hur 060291.
7. Heterodermia microphylla (Kurok.) Skorepa, Lichenologist 8 (2): 132, 1976
Remarks: H. microphylla has labriform soralia and the abundance of erect lobules (mixed with granular soredia) on the upper surface. This species is similar to H. hypoleuca, but differs in having labriform soralia, marginal and laminal lobules, and granular soredia and lobules mix o the upper surface.
SELECTED (OF 36) SPECIMENS EXAMINED: KOREA. Mt. Naejang, alt. 695 m, on rock, 29 June 2003, Hur 030427; Mt. Taebaek, N35°29°46.9” E126°53’40.7°, alt. 1025 m, on rock, 2 November 2003, Hur 030837; Jejudo, N33°21732.3” E126°31°23.5”, alt. 1920 m, on bark, 27 August 2004, Hur 040762; Mt. Jiri, N35°17°25.78” E127°31°59.7°, alt. 1420 m, on Quercus, 17 June 2006, Hur 060173; Mt.Deogyo; N35°51°10.0” E127°44°56.9", alt. 1568 m, on bark, 10 August 2006, Hur 060494; Mt.Baewoon, N35°36’31.3” E127°38°25.67, alt. 1140 m, on Quercus, 17 August 2006, Hur 060605; Mt.Songni, N36°32’39.7” E127°51°49.3”, alt. 700 m, on rock, Hur 060048.
8. Heterodermia obscurata (Nyl.) Trevis., Nuovo Giorn. Bot. Ital. 1: 114, 1869
REMARKS: H. obscurata has labriform soralia, marginal lobules, yellow pigmented lower surface, and by black rhizines. Similar to H. subascendens, but differs in the occurrence of a yellow pigment (which covers almost all the lower surface), the black rhizines, and the striking labriform soralia at the lobe tips.
SPECIMENS EXAMINED: KOREA. Sorokdo, alt. 15 m, on bark, 23 March 2003, Hur 030034; Mt. Gaya, alt. 400 m, on bark, 22 April 2003, Hur 030153; Jejudo, N33°28'25.5” E126°29°47.7°, alt. 145 m, on bark, 26 August 2004, Hur 040623; Jejudo, N33°25’21.9” E126°33'35.7°, alt. 505(614) m, on bark, 26 August 2004, Hur 040680-1; Jejudo, N33°27'15.4” E126°33’41.1’, alt. 370 m, on bark, 29 August 2004, Hur 040863, 040867; Mt. Wonlak, on bark, 18 September 2004, Hur041171; Wando arboretum, 34°21710.3” E126°41°10.9", alt. 535 m, on rock, 13 April 2005, Hur 050136; Mt. Hegsog, N34°41’21.4” E126°40°51.4", alt. 203 m, on rock, 23 September 2005, Hur 050462.
9. Heterodermia pseudospeciosa (Kurok.) W.L. Culb., Bryologist 69: 484, 1967
REMARKS: H. pseudospeciosa is characterized by labriform soralia, lateral branches with subcapitate soralia, and a corticate lower surface. The species is similar to H. speciosa . in morphology, but differs in chemistry. Norstictic and salazinic acids are present in H. pseudospeciosa, while absent in H. speciosa.
SPECIMENS EXAMINED: KOREA. Mt. Jiri, alt. 515 m, on rock, 4 May 2003, Hur 030204; Mt. Baekwoon, alt. 700 m, on rock, 1 June 2003, Hur 030281; Mt. Kem, N34°45’32.3”
Heterodermia in South Korea... 75
E127°59°31.9", alt. 340(310) m, on rock, 15 Febuary 2004, Hur 040033; Baegdam temple, N38°1116.4” E128°21°42.7”, alt. 450 m, on rock, 11 October 2004, Hur 041508; Mt. Hegsog, N34°41'21.4” E126°40’51.4", alt. 203 m, on rock, 23 September 2005, Hur 050451; Mt. Hegsog, N34°41’21.1” E126°40°47.5”, alt. 230 m, on rock, 23 September 2005, Hur 050493; Mt. Juwang, N36°23’58.9” E129°09’53.1”, alt. 310 m, on rock, 16 October 2005, Hur 050618; Mt.Songni, N36°32’41.6” E127°51719”, on Quercus, 10 August 2006, Hur 060025.
10. Heterodermia speciosa (Wulfen) Trevis., Atti Soc. Ital. Sci. Nat. Milano 11: 614, 1868
REMARKS: H. speciosa has capitate soralia on the tips of lobes and lobules, and a lower cortex. The species is similar to H. pseudospeciosa in morphology, but differs in chemistry. Norstictic and salazinic acids are absent in H. speciosa, while present in H. pseudospeciosa.
SPECIMENS EXAMINED: KOREA. Jejudo, alt. 145 m, on bark, 26 August 2004, Hur 040615; Jejudo, N33°28'25.5” E126°29°47.7’, alt. 145 m, on bark, 26 August 2004, Hur 040620; Jejudo, N33°27°15.4” E126°33’41.1’, alt. 370 m, on bark, 29 August 2004, Hur 040849; Mt. Juwang, N36°24’09.6” E129°10’27.0”, alt. 380 m, on rock, 15 October 2005, Hur 050602.
11. Heterodermia subascendens (Asahina) Trass, Folia cryptog. Estonica 29: 20, 1997
REMARKS: H. subascendens is characterized by its maculate upper surface, marginal and grayish white rhizines, an ecorticate yellow lower surface and granular soredia near lobe tips. The species resembles H. hypoleuca in morphology, but differs in the color of rhizines, the presence of a yellow pigment, and the presence of granular soredia on the lower surface. It is also similar to H. hypochraea, differing in the color of rhizines and the presence of granular soredia on the lower surface.
SPECIMENS EXAMINED: KOREA. Sorokdo, alt. 15 m, on bark, 27 December 2003, Hur 030895; Jejudo, N33°28725.5” E126°29°47.7”, alt. 145 m, on bark, 26 August 2004, Hur 040619; Jejudo, N33°27°15.4” E126°33’41.1°, alt. 370 m, on bark, 29 August 2004, Hur 040847.
Earlier reports on further species, not found in the present study
An additional seven species previously recorded from the Korean peninsula but not found in our recent surveys are included in the key according to descriptions provided in the literature.
1) Heterodermia comosa (Eschw.) Follman & Redon, Willdenowia, 6: 446, 1972
Its striking diagnostic character is the presence of long and white cilia on the lobe edges. In Korea, the species has been recorded only once at a single locality on Jeju Island by Kashiwadani et al. (2002). It is widely distributed in tropical regions of Asia and America (Kurokawa 1962).
76 ... Wei & al.
2) Heterodermia dendritica (Pers.) Poelt, Nova Hedwigia 9: 31, 1965
The species is characterized by absence of soredia, often pruinose margins, lower surface ecorticate, white, purple-black only at center, partly yellow pigmented. The taxon is similar to H. propagulifera, but differs in the absence of soredia. Kurokawa (1962) included H. propagulifera as a variety in H. dendritica (as Anaptychia). In Korea, this species has been once recorded from Mt. Jiri by Park (1990).
3) Heterodermia firmula (Nyl.) Trevis., Atti Soc. Ital. Sci. Nat. Milano 11: 615, 1868
The diagnostic character of this species is the yellow medulla. It is close to the endemic Indian species H. albidiflava (Kurok.) D.D. Awasthi, which shows a PD+ deep-yellow reaction (unknown substance). In Korea, H. firmula was recorded only once from Mt. Sobaek by Park (1990). However, we found no species with a yellow medulla on this mountain, although we have many specimens collected there.
4) Heterodermia isidiophora (Nyl.) D.D. Awasthi, Geophytology 3: 114, 1973
Heterodermia isidiophora is a peculiar species in having corticate lower surface and isidia in upper surface. In Korea, the species has been recorded by Park (1990). However, during our study, all the similar specimens with isidia are Anaptychia species, containing no chemical substances, so, whether H. isidiophora really exists in South
Korea still needs further study.
5) Heterodermia loriformis (Kurok.) Swinscow & Krog, Lichenologist 8: 129, 1976
Heterodermia loriformis is a peculiar species in having a white ecorticate lower surface with zones of a yellow pigmentation and white rhizines. Similar to H. dendritica, but differs in its white rhizines. In Korea, the species has been recorded only once by Park (1990) as H. cf. loriformis based on specimens from Mt. Jiri. However, Moon (1999) identified the specimens as H. pandurata. Further studies are still needed to clarify the identity of the taxon.
6) Heterodermia pandurata (Kurok.) J.C. Wei, An enumeration of lichens in China, p. 112, 1991
This species was reported by Moon (1999) on Mts. Jiri and Kaya of Korea. It is very similar to H. hypochraea, but differs in presence of unknown yellow substance (Kurokawa 1962). We didn't find this species during our study.
7) Heterodermia rubescens (Rasanen) D.D. Awasthi, Geophytology 3: 114, 1973
The species is characterized by a corticate lower surface and by mainly marginal rhizines. It had been known only from India until Moon (1999) reported it from Korea. - The voucher specimens are deposited in TNS (Japan), and we found no corresponding specimens in our study.
Heterodermia in South Korea ... 77
Acknowledgments
This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant through Korean National Research Resources Center program (R21-2007-000- 10033-0). We are indebted to Prof. Bruce McCune and Dr. Walter Obermayer for giving valuable comments on the manuscript. The first author also gives her thankfulness to Prof. Jian-Bin Chen, Ms. Mei-Rong Ren, Kwang-Mi Lim and Hae-Sook Jeon for their kind help and cooperation during this study.
Literature cited
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Awasthi DD. 1973. On the species of Anaptychia and Heterodermia from India and Nepal. Geophytology 3: 113-116.
Brodo IM, Sharnoff SD, Sharnoff S$. 2001. Lichens of North America. Yale University Press, New Haven and London, pp. 334-341.
Culberson CF. 1972. Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. J Chromatography 72: 113-125.
Culberson WL. 1967. Chemistry and taxonomy of the lichen genera Heterodermia and Anaptychia in the Carolinas. Bryologist 69: 472-485.
Ekman S. 1999. PCR optimization and troubleshooting, with special reference to the amplification of ribosomal DNA in lichenized fungi. Lichenologist 31: 517-531.
Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes. Application for the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.
Grube M, Arup U. 2001. Molecular and morphological evolution in the Physciaceae (Lecanorales, lichenized Ascomycotina), with special emphasis on the genus Rinodina. Lichenologist 333032772,
Hur JS, Koh YJ, Harada H. 2005. A checklist of Korean lichens. Lichenology 4: 65-95.
Kashiwadani H, Moon KH, Inoue M, Thor G, Kim YS. 2002. Lichens of the Cheju island, Republic of Korea. I. The macrolichens. In Kubodera T., Higuchi M. & Miyawaki (eds.), Proceedings of the 3rd and 4th symposium on collection building and Natural History studies in Asia and the Pacific rim, pp. 115-135, National Science Museum, Tokyo.
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Kurokawa S. 1962. A monograph of the genus Anaptychia. Beihefte Nova Hedwigia 6: 1-115.
Kurokawa S. 1998. A catalogue of Heterodermia. Folia Cryptogamica Estonica 32: 21-25.
Moberg R, Nash TH. 1999. The lichen genus Heterodermia in the Sonoran Desert Area. Bryologist 102: 1-14.
Moon KH. 1999. Lichens of Mt. Sorak in Korea. J. Hattori Bot. Lab. 86: 187-220.
Nordin A, Mattsson JE. 2001. Phylogenetic reconstruction of character development in Physciaceae. Lichenologist 33: 3-23.
Park YS. 1990. The macrolichen flora of South Korea. Bryologist 93:105-160.
Poelt J. 1965. Zur Systematik der Flechtenfamilie Physciaceae. Nova Hedwigia 9: 21-32.
Saitou N, Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4: 406-425.
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Swinscow TDV, Krog H. 1976. The genera Anaptychia and Heterodermia in East Africa. Lichenologist 8: 103-138.
Swofford DL. 2002. PAUP: phylogenetic analysis using parsimony and other methods. Sinauer Associates, Sunderland, Mass.
Wei JC. 1991. An Enumeration of Lichens in China. International Academic Publishers, Beijing.
White FJ, James PW. 1985. A new guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57 (Suppl.): 1-41.
White TJ, Bruns TD, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal DNA genes for phylogenetics. In Innis M.A., Gelfand D.H., Sninsky J.J. & White TJ. (eds), PCR Protocols: a Guide to Methods and Applications, Academic Press, San Diego, pp. 315-321.
Volume 105, pp. 79-87 July-September 2008
Scutellospora striata sp.nov., a newly described glomeromycotan fungus from La Gran Sabana, Venezuela
GISELA CUENCA! & RICARDO HERRERA-PERAZA2‘*
*gcuenca@Ivic.ve
‘Instituto Venezolano de Investigaciones Cientificas (IVIC), Centro de Ecologia Apartado 20632, Caracas 1020-A, Venezuela
*Instituto de Ecologia y Sistematica, CITMA AP 8029, Capdevila, Boyeros, La Habana, Cuba
Abstract — Examination of soil samples from a sclerophyllous shrubland near the town of Iborib6 in La Gran Sabana, Venezuela, revealed an undescribed species of Scutellospora whose spores are ornamented with finger-print-like processes, are ochraceous yellow with a pinkish tint, and have a complex wall structure. The new species, named Scutellospora striata, is the third Scutellospora species described from La Gran Sabana, Venezuela.
Key words — arbuscular mycorrhizal fungus, AMF, Gigasporaceae, taxonomy, tropical species
Introduction
The La Gran Sabana region is located in southeastern Venezuela, South America. It is a highly undulate plain occupying ca. 30.000 km? of a slope from 1450 m to 750 m above sea level directed from the north to the south. This mosaic of igneous and metamorphic rock is one of the earliest shields of the earth crust. It formed the western section of the ancient supercontinent of Gondwana (Schubert & Huber 1989). It was formed in Precambrian age and due to its antiquity and advanced phase of weathering, soils originated from that substrate (Roraima Formation) are mainly sandy with a very small clay fraction and generally very low nutrient content. La Gran Sabana is also the land of the tepuis, which are mountains of vertical walls and flat hilltops (tablelands), and an outstanding region due to the high degree of endemism of its flora (Rull 1991). The plant cover of La Gran Sabana is an intricate mosaic, composed of
+Dr. Herrera-Peraza passed away in December of 2006.
80 ... Cuenca & Herrera—Perazat
numerous types of vegetation. With the exception of the continuous forests at the foot of the eastern tepuis, forests occur in patches encircled by extensive treeless savannas, as well as by shrublands (Dezzeo 1994).
Shrublands are primary plant communities in which the shrub stratum constitutes the principal functional compartment. They consist of a well- developed shrubby stratum 1-3 m high, occasionally with emergent shrubs up to 5 or 7 m tall, with predominantly sclerophyllous leaves (Huber 1994), which almost always grow on rocky sandstone outcroppings or on white quartzitic sands. According to Huber & Febres (2000) they contain a significant number of endemic plants. :
During the inventory of the arbuscular mycorrhizal fungi (AMF) associated with the sclerophyllous shrublands of La Gran Sabana, spores of an undescribed species of the genus Scutellospora with a distinctive ornamentation were found. The fungus is described here as S. striata sp. nov.
Material and methods
Intermittent soil samples were taken during two years below vegetation ina sclerophyllous shrubland situated at Iborib6 (5°36.796’N, 61°29.351’W) in La Gran Sabana. This shrubland grows on sandstone outcrops (Huber 1994). It is highly diverse in plant species, but among the most common are Clusia pusilla, Gongylolepis benthamiana, Euphronia guianensis, Humiria balsamifera, Calliandra sp., Bonnetia sessilis and some small rosettes belonging to the Cyperaceae family such as Rhynchospora barbata and Bulbostylis conifera. Open pot trap cultures started from these soils were maintained in a glasshouse for three months and then dried for one week. The content of the pot was stored at room temperature (~20°C) for almost one year to break latency of spores (Morton et al. 1993). Then, spores were isolated and used to start pure cultures that all failed. Therefore, the description of the species presented below was prepared based on spores isolated only from trap cultures and field soils. The cultures were initiated with Vigna luteola as the host plant, but it died and then was replaced by wild plants germinating from the seed bank. Various very interesting species of Scutellospora spp. were isolated from this place (Herrera-Peraza et al. 2001, Walker et al. 1998). However, no pure culture of any of them could be obtained.
Spores were isolated from the trap pots or from the 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 color chart for British 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 are based on those suggested by Walker (1983) and Walker & Vestberg (1998). Type material has been deposited in the Venezuelan National Herbarium (VEN) and the Cuban National Herbarium, IES-CITMA (HAC).
To study more carefully the external appearance of the spores, a number of them were prepared for Scanning Electron Microscopy (SEM). Prior to the preparation of the material for SEM, spores were put under a dissecting microscope to remove or to
Scutellospora striata sp.nov. (Venezuela) ... 81
break the outer unit component with the aid of fine tweezers. Then, intact or broken spores were rinsed in a phosphate buffer solution and immediately fixed 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 observed for SEM.
Taxonomic description
Scutellospora striata Cuenca & R.A. Herrera sp.nov. Figures 1-3 MycoBAnk MB 511702
Spore singulatim enatae in solo, ochraceae roseo suffusae, superficie nitida, vulgo globosae vel subglobosae, 130-184 x 116-152 um, supra suspensorem bulbosum ochraceum locatae. Sporae parietum structura 6-tunicata. Tunicae prima et secunda respective unitaria et laminata, caeterae omnes membranaceae. Tunica prima 1.5-2 crassa, laevis. Tunica secunda 5-10 um crassa ochracea, in juventute tenuiter laminata et ornata muris 1.0-1.5 um latis, invicem 0.5-0.8 um separatis, striatim dispositis, vestigia digitalia referentibus. Tunica tertia, 0,7 um crassa, rigida, firmiter tunicae secundae adpressa. Tunica quarta, quinta et sexta, flexibles, membranaceae. Tunica quarta difficilis visu, sed Melzeris reactivo dilute rosea evadens. Tunica quinta et sexta firmissime adpressae, solum sub microscopio Normanski obviae, vel sexta Melzeris reactivo purpurea evadens. Scutellum germinativum varie et dense plicatum. Cellulae auxiliares nobis non observatae.
Ho.LotyPe: Slide no. Cuenca 479-4, 13 Feb. 1998, Venezuelan Institute for Scientific
Research.
EryMo_oey Latin striata “with striae’, referring to the spore ornamentation composed of elongated, generally parallel elements separated by grooves.
Spores borne singly in soil, ochraceous yellow of a pinkish tint in color (Ochraceous [8G] to Saffron [49] according to the color chart), having a shinning surface when observed under a dissecting microscope and illuminated with tungsten lamp, generally globose or subglobose (Fig. 1a), 130-184 x 116- 152 um (mean 163 x 152 um, n= 13) with a terminal attached bulbous base, produced from a septate subtending hypha. Bulbous spore base ochraceous yellow in color and 22-29 um wide, with a recurved septum at its base.
SPORE WALL STRUCTURE composed of 6 components or layers generally organized in 3 groups. With the exception of components | and 2, being unit and laminated, respectively, all the remaining components are membranous. Component 1 is contiguous with the sporophore outer wall component, 1.5- 2.0 um thick, and have a smooth surface (Figs. 2a, b and 3). Component 2 is laminated, 5.0-10.0 um thick, and ochraceous yellow (Fig. 3). In young spores the component 2 is finely laminated and ornamented with muri 1.0-1.5 um wide and spaced 0.5-0.8 um apart (Fig. 1b). When seen in a plane view, the muri resemble a finger-print. When spores develop 1 to 3 thick laminae (1.0- 2.0 um thick) originate from component 2 outwards (Figs. 3a-c). Each of these laminae have the same ornamentation as described above. When three laminae
82 ... Cuenca & Herrera—Perazat
are formed, next fine inseparable laminae are synthesized inward. (Fig. 3c). As mentioned before, each thick lamina is ornamented on the upper surface with muri showing a striate pattern. The muri are commonly squared, angular or wave-like in a cross section. The striations rarely follow the same direction on the surface of each of the thick laminae and, thereby, in a plane view the spore ornamentation seems to be composed of tiny squares or rhombs (Fig. 1c). At each two-muri interception wall material is deposited to form a small columella. Therefore, in a cross section the component 2 is composed of up to three lines of rounded cavities resulting from inter-muri and columella inter- spaces (Fig. 2c).
Component 3, measuring approx. 0.7 tm, is a rigid membrane tightly adhered to the lower surface of component 2 or separates from it in vigorously crushed spores (Fig. 1h). Components 4 to 6 are flexible membranes usually tightly adherent to each other.
Component 4 is generally very difficult to discern (Fig. 1f), but its presence becomes evident in Melzer’s reagent because it reacts pale pink (Fig. 1g). Components 5 and 6 are firmly cemented in between and can be discerned only under a microscope equipped with Nomarski interference contrast or in spores crushed in Melzer’s reagent because component 6 becomes readily purple in it. None of these innermost membranes are beaded or amorphous. Germination shield 64 x 72 um in size with complex infolding (Figs. le and 3b).
BULBOUS SPOROPHORE lacks ornamentation. The wall of the sporophore is composed of two layers that are contiguous with components | and 2 of the spore wall (Figure 1d). This wall is 1.5-2.0 um thick, reaching up to 3.0-5.0 um near the spore base. The pore at the attaching point is approximately 1.0 uum diam. Hyphae attached to the sporophore are regularly septated, measuring 5.0-9.0 um diam., and have walls 1.0-2.0 um thick. Auxiliary cells unknown.
DISTRIBUTION AND HABITAT. Known only from La Gran Sabana, Venezuela. Spores of this species have been collected only from the sclerophyllous shrubland at Iboribo and in a treeless savanna dominated by Axonopus canescens, also in the way to the Iborib6 shrubland. Soils are highly acidic, sandy with a very low exchangeable phosphorus and medium level of total N.
MYCORRHIZAL ASSOCIATIONS unknown. Attempts to form mycorrhizae in pure culture have failed, though the species sporulated in a multispecies pot culture with Scutellospora spinosissima, and other unknown species of Acaulospora, Glomus, and Scutellospora.
COLLECTIONS EXAMINED: Type. Isolated from trap culture at the Venezuelan Institute
for Scientific Research, Caracas, Venezuela. Holotype slide no. Cuenca 479-4, 13 Feb. 1998. Isotypes: Cuenca 411-17, March 1997, Cuenca 386-1, 19 Dec. 1996.
Scutellospora striata sp.nov. (Venezuela) ... 83
Figure 1. Scutellospora striata. a) Intact spore mounted in PVLG. b) Ornamentation of spore wall component 2 seen in a plane view. c) Ornamentation of spore wall component 2 of older spore. It resembles rhombs due to the overlapping laminae, each ornamented with striae. d) Non-ornamented bulbous suspensor. e) Germination shield. f) Crushed spore with inner group of membranous components. Most internal components are labeled. g) Heavily crushed spore mounted in PVLG+Melzer’s reagent with stained components 4 and 6. h) Cross view of spore with components 1-6 arranged in two groups.
84 ... Cuenca & Herrera—Perazat
Figure 2. SEM images of S. striata. a) Intact spore with the outermost unit component obscuring the ornamented spore wall component 2. b) The striae on the upper surface of component 2 visible through the broken component 1. c) Cross view of component 2 with lines and round cavities.
Discussion
Scutellospora striata can be readily distinguished from other species in the genus by its peculiar ornamentation. Under a dissecting microscope, the ornamentation is not always evident, because the unit outer component is smooth and usually shines under incident light, though it detaches from the spore easily after low pressure. The presence of the very robust component 1 was evidenced by SEM (Fig. 2a). It has a smooth surface, which is often free of organic debris. However, in some spores lacking the outer component 1, fine soil debris firmly adhere to the spore surface, which show the ornamentation when detached. The ornamentation of component 2 could be visualized under SEM only after manipulating the spore to break the outer unit component (Fig. 2b).
Component 2 is quite complex. Because each layer of the laminated component 2 is ornamented with striae, in a plane view the striae of overlapping layers are visible as tiny squares or rhombs (Fig. 1c). When spores are very young and only one layer (subcomponent) of component 2 is differentiated or when the outermost subcomponent is in focus, the ornamentation resembles a finger-print (Figs. 1b and 3a).
Assessing the number of wall components is very difficult. The number of inner components may be determined when first intact spores are mounted in PVLG for 1-2 hours and crushed by applying pressure to the cover slip to
Figure 3 (on next page). Differentiation of the laminate spore wall component 2 of S. striata. a) Only one lamina of component 2 of young spore. Outer unit component 1 is transparent and easy to see. b) Component | and two laminae of component 2 of older spore. Germination shield is also visible. c) Component | and four laminae of component 2 of mature spore.
Scutellospora striata sp.nov. (Venezuela) ... 85
86 ... Cuenca & Herrera—Perazat
push the internal membranes out (Fig. 1f). Component 4 may also be revealed in spores crushed in PVLG+Melzer’s reagent, because it stains light pink (Fig. 1g).
The sporulation of S. striata in both trap cultures and the field was not abundant. Unfortunately, attempts to grow this fungus in one-species cultures failed and, hence, the properties of its mycorrhizae remain unknown, similar to those of many other unculturable members of the Glomeromycota (Fitter 2005).
Of the described species of the genus Scutellospora, eleven produce ornamented spores. However, while the ornamentation of S. striata spores consists of striae resembling a finger-print when seen in a plane view, that of spores of S. coralloidea, S. dipapillosa, S. gregaria, S. heterogama, S. persica and S. verrucosa is composed of warts, S. crenulata of dome-like subpolygonal papillae separated by pits, S. minuta of spines with round apices, S. nigra of pits, S. nodosa of knobs, and S. spinosissima of blunt spines.
In addition to S. crenulata and S. spinosissima, S. striata is the third newly described species from La Gran Sabana, Venezuela.
Acknowledgments
Fundacite Guayana is gratefully acknowledged for the financial support provided. Professor Bruno Manara from the Venezuelan Botanical Institute prepared the Latin diagnosis and Milagros Lovera, Eduardo Furrazola and Rigel Fernandez gave invaluable comments to the manuscript. We thank Dr. Janusz Blaszkowski and Dr. Marta Cabello for reviewing the manuscript.
Literature cited
Anon. 1969. Royal Botanic Garden Edinburgh. Flora of the British Fungi. Colour Identification Chart. Edinburgh Her Majesty’s Stationery Office.
Dezzeo N. 1994. Ecologia de la Altiplanicie de la Gran Sabana (Guayana Venezolana) I. Scientia Guaianae 4: 1-205.
Fitter AH. 2005. Darkness visible:reflections on underground ecology. Journal of Ecology 93: 231-243.
Herrera-Peraza RA, Cuenca G, Walker C. 2001. Scutellospora crenulata a new species of Glomales from La Gran Sabana Venezuela. Canadian Journal of Botany 79: 674-678.
Huber O. 1994. Los arbustales. Pages 95-106. in N. Dezzeo editor. Ecologia de la Altiplanicie de La Gran Sabana (Guayana Venezolana) I. Scientia Guaianae N° 4.
Huber O, Febres G. 2000. Guia ecoldgica de La Gran Sabana. The Nature Conservancy Caracas. 192 pp. p
Morton JM. 1995. Taxonomic and phylogenetic divergence among five Scutellospora species based on comparative developmental sequences. Mycologia 87: 127-137.
Morton JB, Bentivenga SP, Wheeler WW. 1993. Germ plasm in the international collection of arbuscular or vesicular-arbuscular mycorrhizal fungi (INVAM) and procedures for culture develop- ment documentation and storage. Mycotaxon 48: 491-528.
Scutellospora striata sp.nov. (Venezuela) ... 87
Rull V. 1991. Contribucién a la paleoecologia de Pantepui y la Gran Sabana (Guayana Venezolana) clima biogeografia y ecologia. Scientia Guaianae N° 2:1-132.
Schubert C, Huber O. 1989. The Gran Sabana. Panorama of a Region. Lagoven booklets. Refolit Venezuela.
Sieverding E. 1991. Vesicular-arbuscular mycorrhiza management in tropical agrosystems. GTZ Eschborn Germany.
Walker C. 1983. Taxonomic concepts in the Endogonaceae: spore wall characteristics in species descriptions. Mycotaxon 18: 443-455.
Walker C, Cuenca G, Sanchez F. 1998. Scutellospora spinosissima sp. nov. a newly described Glomalean fungus from low nutrient communities in Venezuela. Annals of Botany 82: 721-725.
Walker C, Vestberg M. 1998. Synonymy amongst the arbuscular mycorrhizal fungi: Glomus claroideum G. maculosum G. multisubstensum and G. fistulosum. Annals of Botany 82:601-624.
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MY COTA ZXON
Volume 105, pp. 89-93 July-September 2008
Arthonia hawksworthii sp. nov. (Ascomycota, Arthoniaceae) on Dimelaena oreina from Turkey
MEHMET GOKHAN HALICI
'mghalici@erciyes.edu.tr Biyoloji Boliimii, Fen Fakiiltesi, Erciyes Universitesi 38039 Kayseri, Turkey
Abstract — Arthonia hawksworthii is described on the thallus of Dimelaena oreina from two localities in Turkey. It is most similar to A. gelidae, a lichenicolous species confined to Placopsis gelida, but mainly differs from this species in that it has ascospores with a gelatinous sheath that stain red in I-Lugol and an epithecium that is light to dark brown with an olivaceous tinge in KOH. The new species is the first Arthonia species reported on Dimelaena.
Key words —fungi, lichens, taxonomy
Introduction
The genus Arthonia Ach. includes c. 500 species, the majority of which are lichenized (Hawksworth et al. 1995). The current concept of the genus is rather broad, as it includes some species with and others without a K/I+ blue ring in the tholus, and the interascal filaments are highly variable within the genus (Coppins 1989, Grube & Matzer 1997). Grube & Matzer (1997) provided useful taxonomical characters for Arthonia species, focusing primarily on lichenicolous taxa.
Material and methods
The type of the new species is deposited in the herbarium at the Erciyes University, Department of Biology, Kayseri. Specimens were examined with an Olympus BH-2 research microscope fitted with Nomarski differential interference contrast optics and a drawing tube. Sections were prepared by hand and examined in I (Lugol’s iodine (MERCK 9261) and Meltzer’s iodine, with [KI] and without [I] pre-treatment with 10% KOH), 10% KOH, and water. Ascospore measurements were taken in water; the extreme values outside the main range are seen in parentheses. The length/breadth (1/b) ratio of the ascospores is provided in the same way.
90 ... Halici
The species
Arthonia hawksworthii Halici, sp. nov. FIGURE 1 MycoBank MB 511724
Fungus in thallo Dimelaenae oreinae incolens. Apothecia nigra, 0.1- 0.4 mm diam. Epithecium 10-20 um altum, olivaceo-brunneum. Hymenium 50-65 um altum, hyalinum. Hypothecium 10-15 ym altum, hyalinum vel pallide brunneum. Asci (23-)27-38 x 14- 18(-22) um, clavati, octospori. Ascosporae 1- septatae, hyalinae, halonatae, (10-)11.5-14 x 4-5.5(-6.5) um, l/b = (1.8-)2.3-3.0(-3.5).
Typus: Turkey, Yozgat, Cayiralan District, El¢i Village, 39°15’N, 35°39’E, alt. 1720 m, on thallus of Dimelaena oreina on siliceous rocks, 14 August 2004, leg. M. G. Halici 0.1386 (herbarium of Erciyes University — holotypus).
EtyMoLocy: Named in honour of David L. Hawksworth, one of the pioneers of
lichenicolous fungi and a tutor of the author. DESCRIPTION: Lichenicolous, on the thalli of Dimelaena oreina, causing slight bleaching, weakly parasitic. AScoMATA apothecia, numerous, dispersed on the surface of the host areoles, black, epruinose, superficial, plane at first, soon becoming + convex, 0.1-0.4 mm diam, 2-4 per areole, lacking an exciple, arthonioid. Epithecium light to dark olivaceous brown, 10-20 um tall, K-, N-; hymenium colourless, 50-65 jm tall, I, + orange-red, KI,,+ slightly blue, I etzer > HYpothecium colourless to pale brown, 10-15 um tall. HAMATHECIUM of paraphysoids, abundant, septate, branched and anastomosed, 2-2.5 um wide; markedly swollen apices, olivaceous brown, to 4-5.5 um wide. AscI broadly clavate, short-stalked, bitunicate in structure, 8-spored, inner part ee red, LSE ca without a So ag blue apical ring, (23-)27-38 x 14-18(-22) uum. ASCOSPORES irregularly arranged in the asci, ellipsoid, hyaline, 1-septate, rounded to somewhat broadly pointed at the apices, slightly constricted at the septa, the upper cell slightly larger than the lower one, with 1-3 oil droplets at times and a gelatinous sheath, the gelatinous sheath I,,+ red, (10-)11.5-14 x 4—5.5(-6.5) um (n = 36), I/b = (1.8-)2.3-3.0(-3.5), all measurements including the closely adhering sheath. CoNIDIOMATA not observed.
ECOLOGY AND DISTRIBUTION: Commensalistic or weakly parasitic on the thalli of Dimelaena oreina as slight bleaching occurs on the infected areoles of the host thalli, especially in the later infection stage. The new species is known from two localities in central and eastern Turkey. As the host species has a wide distribution in the Northern Hemisphere, the species should be sought for elsewhere.
OBSERVATIONS: Arthonia hawksworthii is most similar to A. gelidae R.Sant.,. which was described by Santesson (1986) on the thallus of Placopsis gelida from Sweden. Besides the different hosts, A. hawksworthii differs from A. gelidae in that it has ascospores with a gelatinous sheath that stains red in I wgo, 20d has an epithecium that is light to dark brown with an olivaceous tinge in KOH.
Arthonia hawksworthii sp. nov. (Turkey) ... 91
10 pm.
Fig. 1. Arthonia hawksworthii (holotype). A, mature asci. B, Six ascospore outlines showing the gelatinous sheath, two of the ascospores including oil droplets. Scale = 10 um.
The lichenicolous species of Arthonia characterized by positive iodine reactions of ascospore sheaths include A. pseudopegraphina Matzer, A. intermedia Matzer, A. santessonii Matzer, A. diploiciae Calat. & Diederich, A. almquistii Vain., A. amylospora Almq., A. intexta Almq. and A. obscurior Triebel (Grube & Matzer 1997). The hymenium is also I, + dark blue and KI,,,,+ red in A. gelidae (Santesson 1986), but I, ,,+ only slightly blue and KI,,,+ orange-red in A. hawksworthii. A. epimela Norman, a lichenicolous species on Amandinea punctata, has larger ascomata than A. hawksworthii [0.1-0.4 mm diam vs. 0.4- 0.6 mm diam], narrower ascospores [3.5-4.5 um vs. 4-5.5(-6.5) um] and a brown hypothecium (Almquist 1880, Clauzade et al. 1989). A. epiphyscia Nyl., which is commensalistic or weakly parasitic on Physcia species, has a dark reddish K+ olivaceous hypothecium (Coppins 1992). A. phaeophysciae Grube & Matzer, a parasite causing considerable damage to Phaeophyscia thalli, has asci with a KI + blue apical ring and a K+ greyish hypothecium (Grube & Matzer 1997). A. caerulescens (Almq.) R.Sant., a lichenicolous species confined to the apothecia of Lecanora varia, has shorter ascospores [10-12 um vs. (10-)11.5-14 um] (Almquist 1880). The characters distinguishing the similar species are provided in Table 1.
92 ... Halici
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Arthonia hawksworthii sp. nov. (Turkey) ... 93
ADDITIONAL SPECIMENS EXAMINED: Turkey, Kahramanmaras, Narli District, near the highway to Gaziantep, 37°19’N, 37°08’E, alt. 600 m, on areoles of Dimelaena oreina on siliceous rocks, 7 July 2004, leg. M. G. Halici 0.1387 (herbarium at the Erciyes University).
Acknowledgements
Paul Diederich (Luxembourg) and Violeta Atienza (Spain) are thanked for reviewing this paper. This study was carried out while I was in the Facultad de Farmacia, Universidad Complutense de Madrid, under the direction of David L. Hawksworth, with a grant from TUBITAK. Gokhan Hinisli prepared the figures. I also wish to express my gratitude to Ahmet Aksoy and M. Yasar Dadand1 for their help in the fieldwork.
Literature cited
Almquist S. 1880. Monographia Arthoniarum Scandinaviae. Kongl Svenska Vetensk. Akad. Hand. 17: 1-69.
Clauzade G, Diederich P, Roux C. 1989. Nelikenigintaj fungoj likenlogaj. Bulletin de la Société linnéenne de Provence, numéro spécial 1: 1-142.
Coppins BJ. 1989. Notes on the Arthoniaceae in the British Isles. Lichenologist 21: 195-216.
Coppins BJ. 1992. Arthonia Ach. (1806). In: Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM (eds): The Lichen Flora of Great Britain and Ireland: 74-87. Natural History Museum Publications, London.
Grube M, Matzer M. 1997. Taxonomic concepts of lichenicolous Arthonia species. Bibliotheca Lichenologica 68: 1-17.
Hawksworth DL, Kirk PM, Sutton BC, Pegler DN. 1995. Ainsworth & Bisby’s dictionary of the fungi. 8"" edn. Wallingford.
Santesson R. 1986. Fungi lichenicoli exsiccatae. Fasc. 3-4 (Nos 51-100). Thunbergia 3: 1-18.
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MYCOTAXON
Volume 105, pp. 95-104 July-September 2008
Corticolous lichens of the city of Giresun with descriptions of four species new to Turkey
KENAN YAZICI** & ANDRE APTROOT?
*kcagri_1997@yahoo.com 'Karadeniz Technical University, Faculty of Sciences and Letters Biology Department, Trabzon, Turkey andreaptroot@wanadoo.nl
*Adviesbureau voor Bryologie en Lichenologie G.v.d. Veenstraat 107, NL-3762 XK Soest, The Netherlands
Abstract — Corticolous lichens were studied in the city of Giresun (Turkey). The survey yielded 54 species, all Ascomycota, among which Anisomeridium polypori, Lecanora compallens, Lepraria umbricola and Schismatomma decolorans are new to Turkey. Lecanora compallens is new to Asia while Anisomeridium polypori and Lepraria umbricola are reported for the second time from Asia. Data and accompanying species are given for all taxa while short descriptions are also presented for the four species that are new to Turkey.
Keywords — biodiversity, flora
Introduction
Studies on the lichen flora of Turkey are not as extensive as elsewhere. Last year, many new lichen taxa were reported for Turkey (Dogru & Giiven¢ 2007, Yazici & Aptroot 2007, Yazici et al. 2007a,b). Although there was some previous lichenological research in the Black Sea Coast city of Giresun (Aslan & Yazici 2006, Duman & Yurdakulol 2007, Kinalioglu 2006), several regions in Giresun remained still unexplored. The present paper provides information on corticolous lichens in the city center of Giresun, Turkey.
Materials and methods
The corticolous lichen samples were collected from deciduous trees from 01-30 October 2007 in the city of Giresun (Turkey). The study area is situated at 40°54N, 38°23E. Air-dried lichen samples were observed and studied using stereo and light microscopes. Secondary metabolites were identified by the usual spot tests (Poelt 1974, Purvis et al. 1992, Herk & Aptroot 1999, Wirth 1995, Zedda 2000).
96 ...Yazici & Aptroot
Results
A lichen survey of the city of Giresun yielded 54 species, all Ascomycota, among which Anisomeridium polypori, Lecanora compallens, Lepraria umbricola and Schismatomma decolorans are new to Turkey. Lecanora compallens is new to Asia, while Anisomeridium polypori and Lepraria umbricola are reported here for the second time from Asia. Additionally, Caloplaca flavocitrina and Phaeophyscia chloantha represent second reports for Turkey. Collection data and information accompanying species are given for all lichen taxa, and short descriptions are also presented for the four species new to Turkey. The lichen taxa are listed alphabetically.
Amandinea punctata (Hoftm.) Coppins & Scheid.
City center and Giresun Castle, on Fraxinus sp., Albizia julibrissin, main roadside on Robinia pseudoacacia, accompanying species: Caloplaca obscurella, Candelariella reflexa, Hyperphyscia adglutinata, Lecanora compallens, Phaeophyscia orbicularis and Rinodina gennarii. Yazici 1304.
Anisomeridium polypori (Ellis & Everh.) M.E. Barr
Thallus mostly inconspicuous, effuse, grey or pale grey to green. Perithecia 0.15-0.25 mm diam., subconical to + globose, initially immersed; upper wall with a slightly converted involucrellum. Asci 5-75(-85) x 11-16 um, cylindric- clavate. Ascospores (13-)15-20(-22) x (3.5-)4.5-5 um, 1-3 septate, clavate- fusiform, Pycnidia black, of two types, first one: 100-150 um diam., + sessile, conical, colourless, with ellipsoid or ovoid macroconidia, (3-)3.5-4.5(-5) x 1.8- 2 um, extruded as a white cirrus, in which the conidia are bound by a gelatinous matrix; second one: 50-100 um diam., + immersed, globose, with rod-shaped to narrowly ellipsoid microconidia, 2-3 x 1-1.3 um. Photobiont Trentepohilia. Known throughout Europe, Australia, Canada, China, Costa Rica, Japan,
North America, Russia, Thailand. A second report for Asia.
Giresun: Center; Giresun Castle, 120 m, 40°54’57” N, 38°23’24” E, on Fraxinus sp.,
unpolluted area, accompanying species: Gyalecta truncigena. 10 October 2007. Yazici
1318. ECOLOGY OF THE SURVEY AREA-The area has a typical oceanic climate with a mild and high precipitation prevailing along the coast. It is at 120 m elevation and at 150 m distance from the Black Sea. Annual temperature is 14.2°C in the city of Giresun while annual precipitation is 1305 mm. On average there are 184 rainy days, 6 snowy days and 11 snow-covered days. Winters are more intense with longer snow cover, while the summers are cool. Rainy days occur in every - season, the most rainy days however in autumn and winter. The well-lit site, which is occasionally exposed to strong wind in winter, is rich compared to the forest. It has tree communities with Albizia julibrissin, Robinia pseudoacacia, Salix sp. and Platanus orientalis.
Corticolous lichens of Giresun (Turkey) ... 97
REMARKS—Anisomeridium polypori is a cosmopolitan species frequent suboceanic and mild-temperate region. Generally it especially grows on rough bark of Sambucus, Populus, and Ulmus by rivers and brooks in humid woodland, rarely on shaded rock, and is sometimes common in polluted areas. Anisomeridium polypori is very similar to A. biforme, but it has a less conspicuous thallus, smaller perithecia with a thinner upper wall, and longer and narrower ascospores with often 1-3 septa, but is especially characterized by the long-ostiolate pycnidia with agglutinating conidia.
Arthonia dispersa (Schrad.) Nyl.
City center, on Acer sp., Fraxinus sp., Ficus sp.; Giresun Castle, on Albizia julibrissin, Ficus sp., Fraxinus sp., and Salix sp., accompanying species: Naetrocymbe punctiformis. Yazici 1323.
Arthonia radiata (Pers.) Ach.
Giresun Castle, 120 m, on Robinia pseudoacacia and Fraxinus sp. Yazici 1322.
Bacidia laurocerasi (Duby) Zahlbr.
City center, on Ligustrum sp., Betula sp.; Giresun Castle, 120 m, on Fraxinus sp. Yazici Keay
Caloplaca cerina (Ehrh. ex Hedw.) Th. Fr.
City center and main roadside on Robinia pseudoacacia, accompanying species: Physcia adscendens, P. tenella and Xanthoria parietina. Yazici 1327.
Caloplaca cerinella (Nyl.) Flagey
City center, on Acer sp., accompanying species: Hyperphyscia adglutinata, Lecania fuscella and Lecidella achristotera. Yazici 1301.
Caloplaca citrina (Hoffm.) Th. Fr.
City center, park, on Salix sp., Platanus orientalis, Robinia pseudoacacia, and Ligustrum sp., main roadside on Fraxinus sp. and Robinia pseudoacacia; Giresun Castle, on Albizia julibrissin, accompanying species: Hyperphyscia adglutinata, Phaeophyscia orbicularis, Physcia tenella, P. adscendens and Xanthoria parietina. Yazici 1328.
Caloplaca flavocitrina (Nyl.) H. Olivier
City center and main roadside, on Robinia pseudoacacia, accompanying species: Candelariella reflexa, Lecanora dispersa and Phaeophyscia orbicularis. Found for the second time in Turkey. Yazici 1305.
Caloplaca holocarpa (Hoffm.) A.E. Wade
City center, on Acer sp. and Robinia pseudoacacia, accompanying species: Caloplaca cerina, Hyperphyscia adglutinata, Lecania cyrtella, Lecanora chlarotera, L. dispersa, Lecidella elaeochroma, Physcia adscendens and P. tenella. Yazici 1307.
Caloplaca obscurella (J. Lahm ex K6rb.) Th. Fr.
City center, on Robinia pseudoacacia, accompanying species: Amandinea punctata, Hyperphyscia adglutinata, Phaeophyscia orbicularis and Rinodina gennarii. Yazici 1303.
98 ...Yazici & Aptroot
Candelaria concolor (Dicks.) Stein
City center and main roadside, on Fraxinus sp., accompanying species: Hyperphyscia adglutinata, Lecanora dispersa, Physcia adscendens, P. tenella and Xanthoria parietina. Yazici 1329.
Candelariella aurella (Hoffm.) Zahlbr.
City center and main roadside, on Robinia pseudoacacia, accompanying species: Lecanora dispersa. Yazici 1330.
Candelariella reflexa (Nyl.) Lettau
City center, on Robinia pseudoacacia and Fraxinus sp., accompanying species: Amandinea punctata, Caloplaca flavocitrina, Lecania cyrtella, Lecanora compallens, L. dispersa and Phaeophyscia orbicularis. Yazici 1302.
Chrysothrix candelaris (L.) J.R. Laundon
Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1331.
Evernia prunastri (L.) Ach.
Giresun Castle, 120m, on Albizia julibrissin, accompanying species: Flavoparmelia caperata and Pseudevernia furfuracea. Yazici 1332.
Flavoparmelia caperata (L.) Hale
City center, 5 m, on Pyrus sp., main roadside; Giresun Castle, 120 m, on Robinia pseudoacacia and Albizia julibrissin, accompanying species: Punctelia subrudecta, Parmelia sulcata, Evernia prunastri and Melanelixia fuliginosa. Yazici 1334.
Graphis scripta (L.) Ach.
Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1321.
Gyalecta truncigena (Ach.) Hepp
Giresun Castle, on Fraxinus sp. accompanying species: Anisomeridium polypori. Yazici PS1S:
Hyperphyscia adglutinata (Flérke) H. Mayrhofer & Poelt
City center and along the main roadside 5-20 m, on Tilia sp., Acer sp., Salix sp., Robinia pseudoacacia, Ligustrum sp., Cercis siliquastrum and Fraxinus sp., also Giresun Castle 120 m, on Albizia julibrissin and Platanus orientalis, accompanying species: Amandinea punctata, Caloplaca cerinella, C. holocarpa, C. obscurella, Lecania fuscella, Lecidella achristotera, Phaeophyscia chloantha, P. orbicularis and Rinodina gennarii. Yazici 1303.
Hypogymnia physodes (L.) Nyl. Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Hypogymnia tubulosa, Yazici 1333.
Hypogymnia tubulosa (Schaer.) Hav.
Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Hypogymnia physodes. Yazici 1335.
Corticolous lichens of Giresun (Turkey) ... 99
Lecania cyrtella (Ach.) Th. Fr.
City center, 40 m, on Acer sp., main roadside, on Robinia pseudoacacia, accompanying species: Candelariella reflexa and Lecanora umbrina. Yazici 1316.
Lecania fuscella (Schaer.) A. Massal.
City center, 40 m, on Acer sp., main roadside, on Robinia pseudoacacia, accompanying species: Caloplaca holocarpa, C. cerinella, Hyperphyscia adglutinata, Lecidella achristotera, Physcia adscendens and Lecanora dispersa. Yazici 1307.
Lecania naegelii (Hepp) Diederich & Van den Boom
City center, 40 m, on Acer sp., accompanying species: Lecania cyrtella. Yazici 1336.
Lecanora argentata (Ach.) Malme
City center, 5 m, on Salix sp.; Giresun Castle, 120 m, on Albizia julibrissin, Castanea sp., and Acer sp., main roadside, 5 m, on Ficus sp., accompanying species: Lecidella achristotera. Yazici 1310.
Lecanora campestris (Schaer.) Hue City center, 5 m, on Ficus sp. Yazici 1337.
Lecanora chlarotera Ny.
City center, 5m, on Salix sp. and Acer sp., accompanying species: Caloplaca holocarpa. Vazicw lle
Lecanora compallens Herk & Aptroot
Thallus corticolous, usually 1-3 cm diam., continuous; margin whitish grey, immersed and shiny, usually with low, glaucous grey warts of 0.1-0.2 mm diam., without hypothallus; medulla below soredia 0.1-0.3 mm thick, white. Soralia always present, usually covering most of the thallus, except for a marginal zone of c. 1 mm., 0.1-0.3 mm punctiform openings, soon uprising and aggregating into irregular patches. Soredia granular, in a dense mass, 15- 30 in diam., yellowish to slightly mint-green, contrasting in colour with the thallus. Apothecia and pycnidia unknown. Thallus C- , P- , K-, UV-; soredia C- , P-, K+ yellowish to yellowish brown usnic acid and zeorin found with TLC (Herk & Aptroot 1999).
Lecanora compallens, previously known from Western Europe, most commonly occurs on medium-aged trees, mainly on the flat surfaces of the ridges, and also on acid as well as neutral bark. Usually on the west side of exposed wayside trees, especially Quercus, Fraxinus, Populus, Salix, Sorbus, Tilia, Ulmus and Sambucus.
Known from Belgium, Denmark, England, France, Germany, Lithuania, Luxembourg, Poland, the Netherlands, and Ukraine. New to Asia.
Giresun: Center: Giresun Castle, 40°545°7” N, 38°23’24” E, on Albizia julibrissin, 120 m,
accompanying species: Amandinea punctata, Phaeophyscia orbicularis and Candelariella reflexa. 10 October 2007. Yazici 1302.
100 ... Yazici & Aptroot
ECOLOGY OF THE SURVEY AREA-See Anisomeridium polypori.
REMARKS — Lecanora compallens is very similar to L. expallens and L. barkmaniana, but L. compallens is distinguished from L. expallens in always having a non-sorediate thallus margin of 1 mm with low glaucous grey corticate warts. Furthermore, L. expallens has usually excavate soralia and a more yellowish, never mint-green, colour, and sometimes a black hypothallus is present. L. expallens can be easily separated by the C +orange reaction due to the presence of thiophanic acid. L. barkmaniana and L. compallens have microscopically identical soredia, but L. compallens differs by the yellowish tinge of the soredia and by a much thinner, often mostly immersed thallus, and L. barkmaniana reacts Pd+ yellow.
Lecanora dispersa (Pers.) Sommerf.
City center, on Fraxinus sp., Pyrus sp., and Robinia pseudoacacia, main roadside, on Acer sp., Fraxinus sp., Salix sp., and Robinia pseudoacacia, accompanying species: Caloplaca flavocitrina, C. holocarpa, Candelariella aurella, C. reflexa, Lecania fuscella, Physcia adscendens, Phaeophyscia orbicularis and Rinodina gennarii. Yazici 1309.
Lecanora umbrina (Ach.) A. Massal.
City center and main roadside, 2 m, on Robinia pseudoacacia and Acer sp., accompanying species: Lecania cyrtella. Yazici 1316.
Lecidella achristotera (Nyl.) Hertel & Leuckert
City center and main roadside, on Acer sp., Ficus sp., Fraxinus sp., Pyrus sp.; Giresun Castle, 120 m, on Robinia pseudoacacia, accompanying species: Caloplaca cerinella, Hyperphyscia adglutinata, Lecania fuscella, Lecanora argentata and Lecidella elaeochroma. Yazici 1301.
Lecidella elaeochroma (Ach.) M. Choisy
City center, 10 m, on Acer sp., Ligustrum sp., and Prunus sp., main roadside, 2 m, on Ficus sp. and Robinia pseudoacacia; Giresun Castle, 120 m, on Albizia julibrissin, Laurus sp., and Robinia pseudoacacia, accompanying species: Caloplaca holocarpa and Xanthoria parietina. Yazici 1338.
Lepraria incana (L.) Ach.
Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1339.
Lepraria umbricola Tonsberg
Thallus leprose, powdery appearance, diffuse, deep greenish or whitish greyish, sometimes weft—like, usually thin, rarely thick, cortex absent, lacking marginal lobes and without medulla, protected from direct rain in shaded sites; soredia _ abundant to scattered, forming small granules, granules to 0.1 mm diam and short projecting hyphae frequent. Thamnolic acid present, thallus K + deep yellow, C-, KC-, P + deep orange.
Corticolous lichens of Giresun (Turkey) ... 101
Lepraria umbricola grows on bark, rock, mosses, soil, mostly acidic substrata; shaded, sheltered, humid, sometimes extremely shaded.
Known throughout Europe, Africa, Australia, Canada, Colombia, North America, Pacific Northwest, Russia, and even (probably incorrectly) reported from Thailand. Found for the second time in Asia.
Giresun: Center: Giresun Castle, 40°545’7” N, 38°23'24” E, on Albizia julibrissin, 120 m. 10 October 2007. Yazici 1308. ECOLOGY OF THE SURVEY AREA—See Anisomeridium polypori.
REMARKS-Lepraria umbricola is an oceanic to warm-—temperate species, found on sheltered mossy siliceous rocks and terricolous mosses, sometimes on basal parts of old trunks on shaded sandy soil. Lepraria umbricola is somewhat similar to L. incana and L. eburnea, but L. incana has a more conspicuous medulla. Additionally L. incana has divaricatic acid and zeorine while L. umbricola has thamnolic acid.
Lepraria vouauxii (Hue) R.C. Harris
City center, 20 m, on Tilia sp., main roadside, 2 m, on Magnolia sp. Yazici 1313.
Melanelixia fuliginosa (Fr. ex Duby) O. Blanco et al.
Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Flavoparmelia caperata and Parmelia sulcata. Yazici 1340.
Melanelixia subaurifera (Nyl.) O. Blanco et al.
Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1319.
Naetrocymbe punctiformis (Pers.) R.C. Harris
Giresun Castle, 120 m, on Albizia julibrissin, Fraxinus sp., and Prunus sp., accompanying species: Arthonia dispersa. Yazici 1320.
Opegrapha atra Pers.
City center, 50 m, on Corylus sp., Quercus sp.; Giresun Castle, 120 m, on Acer sp., Corylus sp., and Fraxinus sp. Yazici 1325.
Opegrapha varia Pers.
City center, 2 m, on Acer sp.; Giresun Castle; 120 m, on Acer sp. Yazici 1324.
Opegrapha vulgata (Ach.) Ach. City center, 10 m, on Morus sp. Yazici 1326.
Parmelia sulcata Taylor
City center, main roadside, 2 m, on Fraxinus sp.; Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Flavoparmelia caperata and Melanelixia fuliginosa. Yazici 1341.
Pertusaria amara (Ach.) Nyl. Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1342.
102 ...Yazici & Aptroot
Phaeophyscia chloantha (Ach.) Moberg
City center, main roadside, 2 m, on Magnolia sp. and Robinia pseudoacacia, Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Hyperphyscia adglutinata and Phaeophyscia orbicularis. Found for the second time in Turkey. Yazici 1311.
Phaeophyscia orbicularis (Neck.) Moberg
City center, 5-50 m, on Acer sp., Ligustrum sp., main roadside, 2 m, on Robinia pseudoacacia, Acer sp., Fraxinus sp., and Populus sp.; Giresun castle, 120 m, on Albizia julibrissin, accompanying species: Amandinea punctata, Caloplaca flavocitrina, C. obscurella, Candelariella reflexa, Hyperphyscia adglutinata, Lecanora compallens, L. dispersa, Phaeophyscia chloantha, Physcia tenella, Rinodina gennarii and Xanthoria parietina. Yazici 1302.
Physcia adscendens (Th. Fr.) H. Olivier
City center, 5—50 m, on Acer sp., Fraxinus sp., Ligustrum sp., Populus sp, and Robinia pseudoacacia; Giresun Castle, 120 m, on Albizia julibrissin, Fraxinus sp., Acer sp., accompanying species: Caloplaca cerina, C. holocarpa, Phaeophyscia orbicularis and Physcia tenella. Yazici 1307.
Physcia tenella (Scop.) DC.
City center, 10 m, on Ligustrum sp., main roadside, 2m, on Robinia pseudoacacia and Populus sp., accompanying species: Caloplaca cerina and Physcia adscendens. Yazici 1343.
Punctelia subrudecta (Nyl.) Krog
City center, 2 m, on Pyrus sp., accompanying species: Flavoparmelia caperata. Yazici 1344.
Pseudevernia furfuracea (L.) Zopf
Giresun Castle, 120 m, on Albizia julibrissin, accompanying species: Evernia prunastri and Flavoparmelia caperata. Yazici 1345.
Ramalina farinacea (L.) Ach. Giresun Castle, 120 m, on Albizia julibrissin. Yazici 1346.
Rinodina gennarii Bag.
City center and main roadside, 2 m, on Robinia pseudoacacia, accompanying species: Amandinea punctata, Caloplaca obscurella, Hyperphyscia adglutinata, Lecanora dispersa and Phaeophyscia orbicularis. Yazici 1306.
Schismatomma decolorans (Turner & Borrer ex Sm.) Clauzade & Vézda
Thallus crustose, sorediate, thin, to 3 cm in diam., attached to the substrate with the whole lower surface or with raised, free margins lilac to pinkish grey, — mauve-grey to brown, developing concolorous or slightly paler sorediate patches which usually cover the whole surface of mature specimens, giving a leprose appearance, becoming grey to brownish grey in the herbarium, + rimose-—cracked; a grey-black prothalius often + present; with entire margins
Corticolous lichens of Giresun (Turkey) ... 103
and continuous or more ore less fissured and divided into areoles, flat and mainly parallel to the substrate; upper side clearly different from the underside, not much swollen when wet and usually not wrinkled when dry; photobiont Trentepohlia, algal cells and hyphae surrounded by air. Soredia puntiform, 0.2-0.8(-1) mm daim., + concolorous discrete or sparingly confluent, often covering relatively large areas, mauve-grey to pale lilac-grey, occasionaly with an ochry tinge. Apothecia and pycnidia unknown. Thallus K-, KC-, C-, P-, UV-.
Known throughout Europe (e.g. eastern part of the Italian peninsula, England, France, Germany, Northern Ireland, Spain, Sweden, The Netherlands), Algeria, Cyprus, Iran, North America, Syria, Tanzania. Reported for the second time from Asia.
Giresun: Center; Giresun Castle, 40°545°7” N, 38°23°24” E, on Albizia julibrissin, 120 m, 10 October 2007. Yazici 1314. ECOLOGY OF THE SURVEY AREA-See Anisomeridium polypori.
REMARKS-Schismatomma decolorans is a mild—temperate and suboceanic species, most commonly growing in sheltered and shaded recesses of old rough- barked deciduous trees in wayside and woodland sites, rarely on dry vertical rocks or walls, exposed mineral-rich rock stones and steeply inclined faces in lichen—poor communities, tolerant of relatively high levels of agricultural chemicals and fertilizers.
Xanthoria parietina (L.) Th. Fr.
City center, 2-50 m and main roadside, 2 m, on Ligustrum sp., Acer sp., Elaeagnus sp., Ficus sp., Fraxinus sp., Platanus orientalis, Pyrus sp., Robinia pseudoacacia, and Salix sp., accompanying species: Caloplaca cerina, Candelaria concolor, Lecidella elaeochroma, Physcia adscendens and Phaeophyscia orbicularis. Yazici 1312.
Discussion
The population of Giresun city at the Eastern Black Sea coast is approximately 100,000, according to the census in 2007. The average elevation of the city center is 50 m. Giresun Castle, which is situated in the north of the city, lies at 120 m elevation and at 150 m distance from the Black Sea, in an unpolluted area. Until 2003, sulphur dioxide measurements from the area are not reliable, and these measurements were not performed in 2003—2006. From January 2007 onwards, more reliable measurements have been made. According to these, the annual average sulphur dioxide concentration in the atmosphere is approximately 50ug/m?in 2007 (Turkiye Cumhuriyeti Cevre ve Orman Bakanligi 2007).
In total 33 lichen species, including the four new to Turkey, were found in Giresun Castle, while 30 species were present along the main roads; 36 taxa in the other parts of the city center. All species were found on 20 different
104 ...Yazici & Aptroot
deciduous tree species, such as Acer sp., Albizia julibrissin, Ficus sp., Fraxinus sp., Ligustrum sp., Robinia pseudoacacia and Salix sp.: 26 species were found on Albizia julibrissin, 23 on Robinia pseudoacacia, 18 on Acer sp. and 18 on Fraxinus sp. The area Giresun Castle is apparently an unpolluted area since its 33 lichen taxa include sensitive species such as Evernia prunastri, Pseudevernia furfuracea, Ramalina farinacea, Hypogymnia physodes and H. tubulosa. On the other hand, it is remarkable that still 30 lichen species have been found along the main roads, although these sites are exposed to heavy metals from motor vehicles and other forms of pollution.
Acknowledgements
We are grateful to Professor Orvo Vitikainen and Dr. Leo Spier for linguistic revision and helpful comments on an earlier draft of this manuscript. We also acknowledge the assistance of Dr. Piercey-Normore and Dr. Guttova here.
Literature cited
Aslan A, Yazici K. 2006: Contribution to the lichen flora of Giresun province of Turkey. Acta Bot. Hung. 48: 231-245.
Dogru Z, Giiveng S. 2007: Lichenized and lichenicolous fungi from Bursa province new to Turkey. Mycotaxon 102: 389-394.
Duman DC, Yurdakulol E. 2007: Lichen Records from Saricgicek Mountain in Southern Giresun Province, Turkey. Turk. J. Bot. 31: 357-365.
Herk CM van, Aptroot A. 1999: Lecanora compallens and L. sinuosa, two new overlooked corticolous lichen species from Western Europe. Lichenologist 31: 543-553.
Kinalioglu K. 2005: Lichens of Giresun District Giresun Province, Turkey. Turk. J. Bot 29: 417-423.
Poelt J. 1974: Bestimmungsschliissel Europaischer Flechten. J.Cramer, Lehre.
Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM. 1992: The Lichen Flora of Great Britain and Ireland. Natural History Museum & British Lichen Society, London.
Turkiye Cumhuriyeti Cevre ve Orman Bakanligi. Tiirkiye Hava Izleme Agi, Hava Izleme Raporu, 2007.
Wirth V. 1995: Die Flechten Baden-Wiirttembergs. Teil 1-2. Ulmer, Stuttgart.
Zedda L. 2000: The lichen genera Lepraria and Leproloma in Sardinia (Italy). Cryptogamie Mycol. 21: 249-267:
Yazici K, Aptroot A. 2007: Five lichens new to Turkey. Mycotaxon 100: 21-26.
Yazic1 K, Aptroot A, Aslan A. 2007a: Six lichenized and non-lichenized fungi new to Turkey. Mycotaxon 102: 307-313.
Yazic1 K, Aptroot A, Aslan A. 2007b: Lichen biota of Zonguldak, Turkey. Mycotaxon 102: 257-260.
MYCOTAXON
Volume 105, pp. 105-110 July-September 2008 Balaniopsis triangularis sp. nov. from indoor environments
DeE-WEI Li’, BRYCE KENDRICK’, DAVID SPERO? & CLAIRE MACDONALD4
dewei.li@po.state.ct.us ' The Connecticut Agricultural Experiment Station, Valley Laboratory 153 Cook Hill Road, Windsor, CT 06095
bryce@mycolog.com ’ 8727 Lochside Drive, Sidney, BC, V8L 1M8, Canada
3 Natural Link Mold Lab 4900 Mill Street, Suite 3, Reno, NV 89502
*SanAir Technologies Laboratory 1551 Oakbridge Dr, Suite B, Powhatan, VA 23139
Abstract — A new species of Balaniopsis is described and illustrated from specimens collected during indoor mold inspections in North America.
Key Words — hyphomycete, mitosporic fungi, taxonomy, type
Introduction
In recent years, unidentified triangular spores that appeared to be conidia were recorded by the authors on tape lifts and in spore traps. Occasionally, conidiogenous cells and parts of conidiophore stipes were also seen. On rare occasions, the orientation of the spores relative to the conidiogenous cells and conidiophores was seen. Although the organism has not been successfully grown in culture, a specimen recently collected from conifer wood over a crawl space in a building in Vancouver, Canada, allowed the authors to characterize the species as Balaniopsis triangularis anam. sp. nov..
Materials and methods
Conidiophores and conidia of the fungus were mounted in lacto-fuchsin (0.1 g acid fuchsin, 100 ml 85% lactic acid) (Carmichael 1955). Microscopic
* Corresponding author
106... Li & al.
observations were made using Nomarski differential interference contrast optics. Herbarium acronyms follow Index Herbariorum (Holmgren & Holmgren 1998).
Results
Balaniopsis triangularis D.W. Li & W.B. Kendr., anam. sp. nov. Figures 1-13 MycoBAnkK MB 511446.
Conidiophora macronemata, determinata, erecta, non-ramosa vel ramosa, recta vel flexuosa, lutea, septata, interdum >200 um longa et 0.7-2 um crassa apud basis, 2.9-3.4 um apud apicem. Cellulae conidiogenae polyblasticae, laeves, cupulatae vel obconicae. Conidia unicellularia, triangularia, non catenata, laevia, lutea, 6.0-7.4 x 6.4-8.6 x 3.0- 3.6 um. Teleomorphosis ignota.
Holotypus BPI 878719, Isotypus DAOM 239847 superficie in ligno domi, Vancouver, Canada leg. S. Campbell,. 28 iv. 2008.
EryMoLocy: referring to the triangular shape of the conidia.
CONIDIOPHORES solitary, determinate, erect, unbranched or branched, straight or flexuous, yellowish, smooth, septate, thin-walled when young, becoming thick-walled at maturity, sometimes exceeding 200 um in length, 0.7-2 um wide at base, widening to 2.9-3.4 um at the apex.
CONIDIOGENOUS CELLS terminal, subapical or lateral, obovoid first, becoming cupulate or obconic (champagne flute-shaped) after conidia have been released, due to partial collapse of apical portion, smooth, yellowish, terminal, solitary, (6.8—) 9.3-14.7 (-16.6) (mean = 12 + 2.7, n=17) long, (5.6-) 6.3-7.9 (-8.6) (mean = 7.1 + 0.8 ttm, n=17) wide at the widest point, narrowing to the proximal end (2.6-) 2.7-3.3 (-3.8) (mean = 3.0 + 0.3 um, n=17), subapical or lateral in whorls of 3-5, (7.8—) 8.3-10.5 (-12.6) (mean = 9.4 + 1.1, n = 19) x (5.1-) 6.6-8.4 (-8.9) (mean = 7.5 + 0.9, n = 19). Developing 6-8 conidia polyblastically and perhaps synchronously in the apical area.
Conip1A 1-celled (amerosporous), cuneiform (wedge-shaped) when young, triangular at maturity, yellowish, smooth-walled, later becoming thick-walled, (5.6-) 6.0-7.4 (-8.8) (mean = 6.7 + 0.7, n = 31) x (5.5-) 6.4-8.6 (-9.6) um (mean = 7.5 + 1.1 um, n = 31) x (2.7-) 3.0-3.6 (-4.2) um (mean = 3.3 + 0.3 um, n = 31) um, side wall about 1 um thick, at the three corners 2-2.4 um thick. Conidial secession is rhexolytic. The remnants of separating cell walls persist at the base of some conidia (Figure 7). The corners of the released conidia may appear rounded or, more commonly, truncate. The truncate condition probably indicates some minor collapse of the outer wall subsequent to drying.
TELEOMORPH unknown.
KNOWN GEOGRAPHICAL DISTRIBUTION: British Columbia, Canada; California and New York, USA.
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Balaniopsis triangularis sp. nov. ... 107
20.00 um
F880 um
Figures 1-5. Balaniopsis triangularis. 1. Branched conidiophore, conidiogenous cells, and conidia. 2. Conidiophore with terminal and lateral conidiogenous cells, and conidia. 3, 4. Terminal conidiogenous cells. 5. Terminal and lateral conidiogenous cells. 5. Conidiogenous cells. Scale bars: 1 = 20 um, 2, 5 = 10 um, 3-4 = 5 um.
HABITAT: saprotrophic on damp wood and plywood.
SPECIMENS EXAMINED: CANADA, British Columbia, Vancouver, Marine Drive, crawl space of building, 28 April 2008, Shannon Campbell, holotype (BPI 878719), isotype (DAOM 239847); UNITED STATES, New York, Livingston, 7 March 2007, anonymous, (BPI 878411).
108 ::.. Mer Sal:
5.00 um Figures 6-13. Balaniopsis triangularis. 6. Terminal conidiogenous cells. 7-8. Lateral conidiogenous cells. 9. Lateral conidiogenous cells bearing conidia in face- and side-view
10-12. Conidia in face-view. 13. Conidia in face- and side-view. Scale bars: 6, 8-12 = 5 um, 7, 13 = 10 um.
Discussion
The genus Balaniopsis P.M. Kirk was proposed by Kirk (1985) with a single species, Balaniopsis africana (Kiffer) P.M. Kirk (= Balanium africanum Kiffer). His establishment of the new genus was based on three major characters:
“+.
Balaniopsis triangularis sp. nov. ... 109
1) rhexolytic conidial secession, 2) percurrent extension, and 3) conidia in short, branched chains on unbranched conidiophores. However, Whitton et al. (2002) emended the generic description, reinterpreting the conidiogenous cells and adding a second species. There were significant