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Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius
m y c o s c i e n c e x x x ( 2 0 1 3 ) 1 e9
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Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius Vladimı´r Antonı´n a,*, Rhim Ryoo b, Kang-Hyeon Ka b, Hyeon-Dong Shin c a
Moravian Museum, Department of Botany, Zelny´ trh 6, CZ-659 37 Brno, Czech Republic Department of Forest Products, Korea Forest Research Institute, Seoul 130-712, Republic of Korea c Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Republic of Korea b
article info
abstract
Article history:
Species of Marasmius sect. Marasmius (subsect. Marasmius and Sicciformes) collected in
Received 3 February 2013
South Korea were studied. The detailed morphological descriptions with figures for five
Received in revised form
species and a key for the identification are provided. Marasmius wisteriae is described as a
10 July 2013
new species, and M. bulliardii, M. rotalis, M. tubulatus (all in the subsect. Marasmius), and
Accepted 12 July 2013
M. ruforotula (subsect. Sicciformes) are newly recorded in Korea. Their taxonomic position
Available online xxx
was confirmed by the phylogeny based on the internal transcribed spacer (ITS) and nuclear large ribosomal subunit (LSU) rRNA genes. ª 2013 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Keywords: Euagarics ITS LSU New taxa
1.
Introduction
The section Marasmius of the genus Marasmius (Basidiomycota, Marasmiaceae) contains more than 100 species world-wide (Singer 1986), including the type species of the genus, M. rotula (Scop.: Fr.) Fr. Members of this section form small marasmioid basidiocarps with a membranaceous, radially grooved to sulcate pileus, well-developed collariate lamellae and an insititious filiform stipe. Cheilocystidia and pileipellis cells are in the form of broom cells of the Rotalis or Siccus-type. This group is very complicated taxonomically because of the similarity of macro- and microscopical characters of many taxa, especially in tropical and subtropical regions, is very difficult to obtain exact results without using DNA studies.
This paper represents a continuation of a series of papers dealing with marasmioid and gymnopoid fungi collected in the Republic of Korea (Antonı´n et al. 2009, 2010a,b,c, 2012a,b).
2.
Materials and methods
2.1.
Morphology
Macroscopic descriptions of collected specimens are mostly based on fresh basidiocarps and made by the first and the second author. Microscopic features are described from dried material mounted in H2O, c. 5% KOH, Melzer’s reagent and Congo Red using an Olympus BX-50 light microscope (Tokyo,
* Corresponding author. Tel.: þ420 533435238; fax: þ420 533435313. E-mail addresses: [email protected], [email protected] (V. Antonı´n). 1340-3540/$ e see front matter ª 2013 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.myc.2013.07.003
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Japan). For lamellae, L means number of entire lamellae and l means number of lamellulae tiers between each pair of entire lamellae. For basidiospores, the factors E (quotient of length and width in any one spore) and Q (mean of E-values) are used. Color abbreviations follow Kornerup and Wanscher (1983), herbarium abbreviations Thiers (2013), and the authors of fungal names the Authors of Fungal Names page (http://www. indexfungorum.org/AuthorsOfFungalNames.htm).
2.2.
Molecular phylogeny
Molecular analyses such as DNA extraction, PCR amplification of ITS and LSU rRNA genes and the sequencing were performed according to the methods by Antonı´n et al. (2010a). Phylogenetic analyses were generated in MrBayes version 3.1.2 (Ronquist and Huelsenbeck 2003), applying a GTR þ I þ G substitution model with 1 106 generations, saving a tree every 10,000th generation, with the first 1000 trees discarded as burn-in for ITS and LSU datasets. The two analyses were performed atleast four times and were combined in a 50% majority rule consensus tree to obtain estimates for the posterior probabilities (PPs) by Antonı´n et al. (2010b). ITS and LSU dataset of M. siccus were chosen as an outgroup for rooting purpose based on a study by Tan et al. (2009) and Wannathes et al. (2009). The alignment datasets were deposited in TreeBASE (http://www.treebase.org) under ID S14462. For the list of specimens studied, including GenBank accession numbers, see Table 1.
3.
Results and discussion
3.1.
Taxonomy
Marasmius subsect. Marasmius Synonyms: Marasmius subsect. Pararotulae (Singer) Singer (type species: M. pararotula Singer). Marasmius subsect. Penicillati Singer (type species: M. graminum (Lib.) Berk.).
Type species: Marasmius rotula (Scop.: Fr.) Fr. Species are characterized in having pileipellis consisting of broom cells of the Rotalis-type. Marasmius wisteriae Antonı´n, R. Ryoo & H.D. Shin, sp. nov. Figs. 1A, 2. MycoBank no.: MB 803299. Pileus brownish orange to whitish, central dot dark brown or black. Basidiospores 11.5e13.5 2.5e3.2 mm, clavatefusoid, fusoid. Cheilocystidia and pileipellis cells of the Rotalis-type. Holotype: Republic of Korea, Chuncheon, Kangwon University experimental forest, 15. VI. 2009 leg. V. Antonı´n and R. Ryoo (BRNM 718761). rRNA gene sequences ex-holotype: JN003839 (ITS), JN003844 (LSU). Etymology: wisteriae, found on leaves of Wisteria floribunda. Pileus up to 2.5 mm broad, convex-conical with central umbilicus with dark brown or black dot, striateesulcate with crenulate, inflexed to straight margin, finely tomentose, pale brownish orange (paler than 6C4) to whitish. Lamellae distant, L ¼ 8e12, l ¼ 0, collariate, pale cream with concolorous, finely pubescent edge. Stipe up to 30 mm long, glabrous, lustrous, concolorous with lamellae and translucent at apex, brown, black-brown towards base. Basidiospores 11.5e13.5 (e14) 2.5e3.2 mm, 12.8 2.9 mm on average, E ¼ 3.8e5.2, Q ¼ 4.45, clavate-fusoid, fusoid, narrowly lacrimoid. Basidia (only one found) 21 8.0 mm, 4-spored, clavate. Basidioles 15e26 (2.5e) 4.0e8.0 mm, clavate, fusoid. Cheilocystidia similar to pileipellis broom cells, 12e20 7.0e13 mm, (broadly) clavate to pyriform, subvesiculose, diverticulate, thin-walled. Trama hyphae cylindrical, thin-walled, up to 10 mm wide. Pileipellis a hymeniderm composed of 14e25 10e15 mm, clavate, pyriform to subvesiculose cells of the Rotalis-type, thin-walled; diverticula digitate, thick-walled, obtuse. Stipitipellis a cutis of cylindrical, slightly thick-walled, smooth, up to 5.0 (e6.0) mm wide hyphae
Table 1 e Data on Marasmius specimens used to this study. Species
Marasmius bulliardii agg. M. bulliardii M. rotalis agg. M. rotalis agg. M. rotalis agg. M. rotula M. rotula M. rotula M. ruforotula M. ruforotula M. siccus M. siccus M. tubulatus M. wisteriae M. wisteriae
Locality
Korea Hungary Cameroon Korea Korea Germany Denmark USA Korea Korea Korea Russia Korea Korea Korea
Specimen ID
Ryoo KG 154, ¼BRNM 724476 Antonı´n 06.160, ¼BRNM 705006 Antonı´n Cm 01.107, ¼BRNM 666156 Ryoo KG 132, ¼BRNM 724479 Ryoo KG 232, ¼BRNM 724477 GLM 45962 Landvik NN005958 PBM 2563 Antonı´n 07.158, ¼BRNM 714674 Ryoo KG 107, ¼BRNM 714676 Ryoo KG 028, ¼BRNM 714680 Landvik NN050245 Antonı´n 07.160, ¼BRNM 714675 Antonı´n 09.102, ¼BRNM 718761 Ryoo KG 223, ¼BRNM 724478
GenBank accession number ITS
LSU
No data JN540056 KC415763 JN003837 KC415765 No data JN943598 DQ182506 FJ936150 FJ936152 FJ904985 JN943596 FJ936151 JN003839 JN003838
JN003846 JN540057 KC415764 JN003843 No data AY207238 JN941146 No data FJ917612 FJ917614 FJ904980 JN941142 FJ917613 JN003844 No data
Species names in bold type were retrieved from GenBank. The others were sequenced in this study. The herbarium abbreviations follow Thiers (2013).
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 1 e Basidiocarps of Marasmius species from South Korea. A: M. wisteriae (Ryoo KG 223). B: M. tubulatus (Antonı´n 07.160). C: M. rotalis (Ryoo KG 232). D: M. bulliardii (Ryoo KG 154). E: M. ruforotula (Antonı´n 07.158).
with brown walls in KOH. Clamps present. Chemical reactions: Trama, stipe medulla and stipitipellis hyphae non-dextrinoid or slightly dextrinoid, other structures non-dextrinoid. Habitat: On leaves of Wisteria floribunda, and on dead fallen leaves. Specimens examined: Republic of Korea, Chuncheon, Kangwon University experimental forest, alt. c. 1080 m, coord. 37 480 48.6900 N, 127 510 04.1400 E, 15 July 2009 leg. V. Antonı´n (09.102) and R. Ryoo (holotype, BRNM 718761); Seoul, Bukhansan National Park, in the vicinity of Jeongreung stream, 13 July 2008 leg. R. Ryoo (KG 223) (BRNM 724478). ? Japan, Miyazaki, Miyazaki College of Agri-culture, 29 June 1939 leg. Y. Arataka (K(M) 171077, as M. rotula). Marasmius wisteriae is especially characterized by a pale colored pileus with a dark central dot, distant lamellae, and large, clavate-fusoid, fusoid, narrowly lacrimoid basidiospores. Such combination of characters is not published in any described species. Marasmius pandoanus Singer has a white pileus and larger basidiospores (14.5e16.5 3.7e4.6 mm); M. oaxacanus Singer has also a white pileus, more close lamellae (L ¼ 10e16) and larger basidiospores (10e14 3e4 mm); M. arimanus Dennis differs by colored lamella edge and larger basidiospores (11e14 3.5e4 mm) (Singer 1976). A microscopically very similar specimen from Japan has been found in the K(M) herbarium. Its macroscopic description is not available, therefore, the exact identification is not possible (mentioned with a question mark in the list above).
Marasmius tubulatus Petch, Trans. Br. mycol. Soc. 31: 42, 1948. Figs. 1B, 3. Pileus up to 3 mm broad, convex-conical with obtuse center, then convex or almost applanate with slightly depressed center and inflexed margin, finely tomentose, entirely striateesulcate with crenulate margin, uniformly light brown to brownish orange (6C4). Lamellae distant, L ¼ 6e7, whitish, then dirty pale cream, with concolorous, later granulose-colored (pileus color) edge. Stipe up to 10 mm long, glabrous, brown (6E5e6) to black-brown with paler apex, originating from substrate. Rhizomorphs smooth, black-brown. Basidiospores (collapsed, only a few found) (7.0e) 8.0e10 3.75e4.5 mm, 9.0 4.0 mm on average, E ¼ 1.9e2.4, Q ¼ 2.2, ellipsoid-fusoid or subfusoid. Basidioles 12e20 4.0e9.0 mm, clavate, fusoid. Cheilocystidia of the Rotalis-type, 15e19 8.0e11 mm, (broadly) clavate to pyriform, thin-walled, with pale (yellow) gray-brown walls in KOH. Pileipellis a hymeniderm composed of 12e25 10e14 mm, clavate to pyriform cells of the Rotalistype, thin-walled; diverticula up to 3.0 1.0 mm, digitate, often irregular; thick-walled parts (yellow) gray-brown in KOH. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps present. Chemical reactions: Trama, stipitipellis and stipe medulla hyphae (slightly) dextrinoid, other structures nondextrinoid. Habitat: On dead twigs of Salix sp. Specimen examined: Republic of Korea, Namyangju, Korea University experimental farm, alt. c. 140 m, 37 340 5200 N, 127
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 2 e Marasmius wisteriae (BRNM 718761). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
140 4000 E, 30 July 2007 leg. V. Antonı´n (07.160), H.D. Shin, R. Ryoo and M.J. Park (BRNM 714675). Our collection of M. tubulatus is characterized by a small uniformly light brown to brownish orange pileus, very distant lamellae (L ¼ 6e7) with a concolorous or granulose-colored edge, a short stipe not originating from rhizomorphs, moderately large basidiospores, and pileipellis cells of the Rotalis-type. Marasmius tubulatus has been mentioned by various authors in various senses with different character combinations. A description by Manimohan and Leelawathy (1989) is the closest one to our fungus. Description by Tan et al. (2009) differs by slightly closer lamellae (L ¼ 10e13) and larger cheilocystidia (12e27 7e18 (e22) mm).
Marasmius bulliardii agg. Que´l., Bull. Soc. bot. Fr. 24: 323, 1878. Fig. 1C. Pileus conical, then convex with depressed center and central dot or small papilla in it, sulcate, ochraceous brown, whitish to pale brownish at center. Lamellae distant, L ¼ 15e17, pale cream, then darker, with concolorous edge. Stipe glabrous, dark brown to black-brown. Rhizomorphs probably absent. (Description based on dry basidiocarps and photographs.) Basidiospore (only one found) 8.5 5.5 mm, E ¼ 1.55, ellipsoid. Hymenium damaged. Pileipellis a hymeniderm of 20e35 (e45) 17e25 mm, broadly clavate or vesiculose broom cells of the Rotalis-type, thin- to slightly thick-walled; projections up to 3.0 1.5 mm; thickwalled parts of cells and projections brown in KOH.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 3 e Marasmius tubulatus (BRNM 714675). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Stipitipellis a cutis of cylindrical, up to 4.0 mm wide hyphae with dark brown walls in KOH. Clamps present. Chemical reactions: Stipitipellis and stipe medulla hyphae (slightly) dextrinoid at apex, other structures non-dextrinoid. Habitat: On dead leaves. Specimen examined: Republic of Korea, Deogyusan National Park, Cheon-yeon falls, 24 Aug. 2007 leg. R. Ryoo (KG 154) (BRNM 724476). The characters agree with M. bulliardii, and LSU sequences are also similar to the European taxon from Hungary (Table 1, Fig. 7). Microscopically, it differs only by larger pileipellis cells. However, material is very poor, a macroscopic description of fresh basidiocarps is absent, and, moreover, hymenium is damaged. Therefore, it is necessary to re-collect it to settle its taxonomic status. Marasmius wettsteinii Sacc. & P. Syd. published by Cho and Yoo (1999) from the Republic of Korea may represent our collection. However, the authors mentioned very small basidiospores (5e6 3e4 mm).
Marasmius rotalis agg. Berk. & Broome, J. Linn. Soc., Bot. 14: 40, 1873. Figs. 1D, 4. Pileus conical with depressed center and dark central dot, sulcate, finely tomentose, pale greyish with slightly paler margin. Lamellae distant, L ¼ 13e16, white, with concolorous edge. Stipe glabrous, black-brown. Rhizomorphs (probably) absent. (Description based on dry basidiocarps and photographs.) Basidiospores 7.5e9.0 (4.25e) 4.5e5.0 mm, 8.4 4.7 mm on average, E ¼ 1.6e2.0, Q ¼ 1.8, ellipsoid. Basidioles 15e29 4.0e7.0 mm, clavate, cylindrical or fusoid. Cheilocystidia 11e24 7.0e15 mm, of the Rotalistype, clavate, (sub)vesiculose, hyaline. Pileipellis a hymeniderm of (12e) 15e25 (e30) 8.0e20 mm, pyriform, broadly clavate or subvesiculose broom cells of the Rotalis-type, thin- to slightly thick-walled at apex. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps present. Chemical reactions: Stipitipellis and stipe medulla hyphae non-dextrinoid or weakly dextrinoid, other structures non-dextrinoid.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 4 e Marasmius rotalis (BRNM 724477). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Habitat: On dead leaves (e.g., Quercus). Specimen examined: Republic of Korea, Pyeongchang, Chungtaesan Resort Forest, 17 July 2008, leg. R. Ryoo (KG 232) (BRNM 724477). Our collection agrees with descriptions of M. rotalis (e.g., Antonı´n 2007), however, Singer (1976) described it without a dark central dot. Desjardin and Horak (1997) mentioned only 7.5e9 3e3.5 mm large basidiospores. The type specimen of M. rotalis (Sri Lanka, Peradeniya, K(M)92650!) differs by having slightly narrower basidiospores (7.0e10 3.5e 4.5 mm) and smaller pileipellis broom cells ((12e) 15e17 5.0e12 mm). Another very similar South-Korean collection (Chuncheon, Dongsan-myeon, Bongmyong-ri, 20 Aug. 2007 leg. R. Ryoo KG 132, BRNM 724479) forms, according to LSU and ITS, the same tree with the collection of M. rotalis from Cameroon (prov. Sud, Dja Biosphere Reserve, Nkoelgnegue
near Ekom, ca. 37 km to E of Somalomo, 12 Apr. 2001 leg. V. Antonı´n, BRNM 666156). However, the Korean collection is too poor for detailed microscopic studies. Both above mentioned collections are close in comparison of LSU sequences with two sequences of M. rotula taken from GenBank. However, both species groups distinctly differ by their macroscopic characters. Marasmius rotalis species complex is very complicated in East and South-East Asia (cf., Tan et al. 2009; Wannathes et al. 2009). Marasmius subsect. Sicciformes Antonı´n, Acta Mus. Moraviae, Sci. nat., 76: 145. 1991. Type species: Marasmius curreyi Berk. & Broome Species are characterized in having pileipellis consisting of broom cells of the Siccus-type. Marasmius ruforotula Singer, Sydowia 2: 34, 1948. Figs. 1E, 5.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 5 e Marasmius ruforotula (BRNM 714676). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Pileus 1e7 mm broad, convex-hemispherical or conical, then (flat-)convex, with depressed center and small papilla, sulcate, finely tomentose, dark brown or reddish brown (7e8BeD6e8), darker reddish brown (8E8 to 9CeDeE8) at center, paler (7e8C5e6) when old, paler in striae to less distinctly striped, almost cream-whitish at very margin and in oldest (revived?) basidiocarps. Lamellae distant, L ¼ 9e11, slightly intervenose when old, pale cream, later yellowish white (3e4A2), with colored (pileus color) edge. Stipe up to 25 mm long, glabrous, dark brown to black-brown (7EeF6e7). Rhizomorphs present. Basidiospores 7.5e8.5 (e9.5) 4.0e5.0 mm, 8.4 4.5 mm on average, E ¼ 1.5e2.1, Q ¼ 1.8e1.9, ellipsoid, ellipsoid-fusoid. Basidia 23e30 7.0e10.5 mm, 4-spored, clavate. Cheilocystidia of the Siccus-type, 11e19 8.0e12 mm, (broadly) clavate to pyriform, thin-walled with thick-walled apex, mixed with scattered smooth, clavate, thin-walled, regular to irregular cells with brown walls in KOH. Pileipellis a hymeniderm of (9.0e) 12e18 7.0e11 mm, clavate, pyriform or vesiculose, often irregular broom cells of the Siccus-type, thin-walled with slightly thick-walled apex; diverticula 4e20, up to 6.0 1.0 mm, cylindrical to conical; mixed with irregular, thickwalled cells (almost) without projections; thick-walled parts (reddish) brown in KOH. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps
present. Chemical reactions: Stipitipellis, pileus context and stipe medulla hyphae (slightly) dextrinoid, other structures non-dextrinoid. Habitat: On dead twigs of a broadleaved tree (Salix sp.?) and on dead stems of grass and Juncus sp. Specimens examined: Republic of Korea, Namyangju, Korea University experimental farm, alt. c. 140 m, 37 340 5200 N, 127 140 4000 E, 30 July 2007 leg. V. Antonı´n (07.158), H.D. Shin, R. Ryoo and M.J. Park (BRNM 714674); Yangpyeong, Jungmisan National Resort Forest, 37 340 4600 N, 127 270 0700 E, alt. c. 395 m, 1 July 2008 leg. V. Antonı´n (08.47) and R. Ryoo (BRNM 718685); Guri, Dongguneung (East Nine Tombs), alt. 30 m, 37 370 0300 N, 127 080 0100 E, 16 Aug. 2007 leg. R. Ryoo (KG 107) (BRNM 714676). In literature M. ruforotula has been mentioned in the several senses (e.g., Singer 1976; Pegler 1983; Desjardin and Horak 1997). They especially differ each other by the basidiospore size. Our collections fit to the description by Desjardin et al. (2000) from Indonesia. The Yangpyeong collection (BRNM 718685) differs by the presence of a dark central pileus dot, slightly more close lamellae (L ¼ 11e13), an up to 40 mm long stipe, and the presence of smooth cells among typically developed cheilocystidia.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 6 e Phylogenetic tree of Marasmius sect. Marasmius inferred from Bayesian analysis of the ITS region (ITS1-5.8S-ITS2), showing mean branch lengths of a 50% majority rule consensus tree, obtained from an MCMC analysis of 106 generations. Species names in bold type were retrieved from GenBank.
Fig. 7 e Phylogenetic tree of Marasmius sect. Marasmius inferred from Bayesian analysis based on the LSU rRNA gene sequences, showing mean branch lengths of a 50% majority rule consensus tree, obtained from an MCMC analysis of 106 generations. Species names in bold type were retrieved from GenBank.
3.2. Key to the species in Marasmius sect. Marasmius found in the Republic of Korea 1 Cheilocystidia and pileipellis broom cells in the form of Siccus-type (subsect. Sicciformes) ........................................................................................... M. ruforotula 1 Cheilocystidia and pileipellis broom cells in the form of Rotalis-type (subsect. Marasmius) ................................................................................................................. 2 2 Basidiospores 12–13.5(–14) × 2.5–3.25 µm, fusoid, narrowly lacrimoid M. wisteriae 2 Basidiospores 7.0–10 × 3.75–5.0 µm, ellipsoid to ellipsoid-fusoid ........................... 3 3 Pileus ochraceous brown except for whitish centre; lamellae moderately distant (L = 15–17); basidiospore 8.5 × 5.5 µm, E = 1.55; pileipellis cells 20–35(–45) × 17–25 µm; on dead leaves ................................................................................... M. bulliardii 3 Pileus pale greyish or light brown to brownish orange; basidiospores 2.5–4.5 µm broad, Q = 1.8–4.5 ....................................................................................................... 4 4 Pileus light brown to brownish orange; lamellae distant (L = 6–7); basidiospores (7.0–)8.0–10 × 3.75–4.5 µm, Q = 2.2; on twigs ...................................... M. tubulatus 4 Pileus pale greyish; lamellae more close (L = 13–16); basidiospores 7.5–9.0 × 4.5– 5.0 µm, Q = 1.8; on dead leaves .................................................................... M. rotalis
3.3.
Phylogenetic analysis
Phylogenetic trees of the genus Marasmius sect. Marasmius were inferred from MCMC analyses based on ITS and LSU rRNA gene
sequences obtained in this study and retrieved from GenBank (Figs. 6, 7). For ITS and LSU rRNA gene dataset, 13 and 12 sequences were aligned and trimmed to 514 and 897 base pairs at each other. The analyzed sequences are divided in two groups following subsections in this topology. The one group consisted of subsect. Marasmius and the other included M. ruforotula (subsect. Sicciformes). The results are concordant with the delimitations of five species based on macro- and micromorphological characteristics. Marasmius wisteriae forms a distinct clade. Marasmius rotalis formed the same subclade with sequences of M. rotula from GenBank, type species of the genus Marasmius in both datasets (see remarks in M. rotalis). Although two specimens of this species were collected in Korea, they represented the distinct phylogenetic position in ITS dataset. One of them, BRNM 724479, formed a grouping with Cameroon in LSU as well as ITS of phylogenetic tree. We excluded samples BRNM 724477 and 724478, from the phylogenetic analysis of LSU rRNA genes owing to its amplifications of multiple fragments. Because a very small quantity of DNA was extracted from M. bulliardii, BRNM 724476, we couldn’t obtain ITS sequences of this isolate and be included in a phylogeny tree. LSU sequences of
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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this isolate, BRNM 724476, however, were similar to those of M. bulliardii collected from Hungary. They were different 3 out of 918 nucleotide characters. Phylogenetic analysis of ITS and LSU rRNA genes for Marasmius sect. Marasmius suggested the independent taxonomic position of morphologically similar species, although they were limited by the absence of sequences of several isolate.
4.
Conclusions
Studies of South-Korean Marasmius sect. Marasmius specimens confirmed the presence of five taxa till now. Macro- and microscopical characters of some species of this section (see e.g., discussion on M. bulliardii and M. rotalis in this study) are very similar to almost identical, and it is very difficult to obtain exact taxonomic results without using DNA studies. Moreover, it seems that many Marasmius species have their morphologically very similar vicariant taxon in other geographical areas (Antonı´n unpublished studies, Petersen pers. comm.).
Acknowledgements The collecting trip to the Republic of Korea and the studies of the material by the first author were supported by the Czech Science Foundation (No. 206/07/J003), and due the support provided to the Moravian Museum by the Ministry of Culture of the Czech Republic as part of its long-term conceptual development programme for research institutions (ref. MK000094862). The other authors were supported by the Korea Research Foundation Grant funded by the Korean Government (KRF-2006-F00001) and by project (FP 0801-2009) of Korea Forest Research Institute.
references
Antonı´n V, 2007. Monograph of Marasmius, Gloiocephala, Palaeocephala and Setulipes in tropical Africa. Fungus Flora of Tropical Africa 1: 1e177. Antonı´n V, Ryoo R, Ka KH, Shin HD, 2012a. Marasmioid and gymnopoid fungi of the Republic of Korea. 5. Marasmius sect. Hygrometrici. Mycotaxon 119: 405e419; http://dx.doi.org/ 10.5248/119.405.
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Antonı´n V, Ryoo R, Shin HD, 2009. Marasmioid and gymnopoid fungi of the Republic of Korea. 1. Three interesting species of Crinipellis (Basidiomycota, Marasmiaceae). Mycotaxon 108: 429e440. Antonı´n V, Ryoo R, Shin HD, 2010a. Marasmioid and gymnopoid fungi of the Republic of Korea. 2. Marasmius sect. Globulares. Persoonia 24: 49e59; http://dx.doi.org/10.3767/003158510X496107. Antonı´n V, Ryoo R, Shin HD, 2010b. Marasmioid and gymnopoid fungi of the Republic of Korea. 3. Two new taxa of Marasmius sect. Sicci with caulocystidia and/or setae. Mycotaxon 111: 369e377. Antonı´n V, Ryoo R, Shin HD, 2010c. Two new marasmielloid fungi widely distributed in the Republic of Korea. Mycotaxon 112: 189e199. Antonı´n V, Ryoo R, Shin HD, 2012b. Marasmioid and gymnopoid fungi of the Republic of Korea. 4. Marasmius sect. Sicci. Mycological Progress 11: 615e638; http://dx.doi.org/10.1007/ s11557-011-0773-y. Cho DH, Yoo ID, 1999. Notes on Korean higher fungi (XV). Plant Research 2: 133e138. Desjardin DE, Horak E, 1997. Marasmius and Gloiocephala in the South Pacific Region: Papua New Guinea, New Caledonia, and New Zealand taxa. Part 1: Papua New Guinea and New Caledonia taxa, Part 2: New Zealand taxa. Bibliotheca Mycologica 168: 1e152. Desjardin DE, Retnowati A, Horak E, 2000. Agaricales of Indonesia. 2. A preliminary monograph of Marasmius from Java and Bali. Sydowia 52: 92e194. Kornerup A, Wanscher JH, 1983. Methuen handbook of colour, 3rd edn. Methuen Co., London. Manimohan P, Leelawathy KM, 1989. Marasmius species new to India. Sydowia 41: 185e199. Pegler DN, 1983. Agaric flora of the Lesser Antilles. Kew Bulletin Additional Series 9: 1e668. Ronquist F, Huelsenbeck JP, 2003. MRBAYES 3: Bayesian phylogenetic inference under mixed molds. Bioinformatics 19: 1572e1574. Singer R, 1976. Marasmieae (Basidiomycetes e Tricholomataceae). Flora Neotropica 17: 1e348. Singer R, 1986. The Agaricales in modern taxonomy, 4th edn. Koeltz, Koenigstein. Tan Y-S, Desjardin DE, Perry BA, Vikineswary S, Noorlidah A, 2009. Marasmius sensu stricto in Peninsular Malaysia. Fungal Diversity 37: 9e100. Thiers B, 2013 [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 5 Jan 2013. Wannathes N, Desjardin DE, Hyde KD, Perry BA, Lumyong S, 2009. A monograph of Marasmius (Basidiomycota) from Northern Thailand based on morphological and molecular (ITS sequences) data. Fungal Diversity 37: 209e306.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius Vladimı´r Antonı´n a,*, Rhim Ryoo b, Kang-Hyeon Ka b, Hyeon-Dong Shin c a
Moravian Museum, Department of Botany, Zelny´ trh 6, CZ-659 37 Brno, Czech Republic Department of Forest Products, Korea Forest Research Institute, Seoul 130-712, Republic of Korea c Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Republic of Korea b
article info
abstract
Article history:
Species of Marasmius sect. Marasmius (subsect. Marasmius and Sicciformes) collected in
Received 3 February 2013
South Korea were studied. The detailed morphological descriptions with figures for five
Received in revised form
species and a key for the identification are provided. Marasmius wisteriae is described as a
10 July 2013
new species, and M. bulliardii, M. rotalis, M. tubulatus (all in the subsect. Marasmius), and
Accepted 12 July 2013
M. ruforotula (subsect. Sicciformes) are newly recorded in Korea. Their taxonomic position
Available online xxx
was confirmed by the phylogeny based on the internal transcribed spacer (ITS) and nuclear large ribosomal subunit (LSU) rRNA genes. ª 2013 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Keywords: Euagarics ITS LSU New taxa
1.
Introduction
The section Marasmius of the genus Marasmius (Basidiomycota, Marasmiaceae) contains more than 100 species world-wide (Singer 1986), including the type species of the genus, M. rotula (Scop.: Fr.) Fr. Members of this section form small marasmioid basidiocarps with a membranaceous, radially grooved to sulcate pileus, well-developed collariate lamellae and an insititious filiform stipe. Cheilocystidia and pileipellis cells are in the form of broom cells of the Rotalis or Siccus-type. This group is very complicated taxonomically because of the similarity of macro- and microscopical characters of many taxa, especially in tropical and subtropical regions, is very difficult to obtain exact results without using DNA studies.
This paper represents a continuation of a series of papers dealing with marasmioid and gymnopoid fungi collected in the Republic of Korea (Antonı´n et al. 2009, 2010a,b,c, 2012a,b).
2.
Materials and methods
2.1.
Morphology
Macroscopic descriptions of collected specimens are mostly based on fresh basidiocarps and made by the first and the second author. Microscopic features are described from dried material mounted in H2O, c. 5% KOH, Melzer’s reagent and Congo Red using an Olympus BX-50 light microscope (Tokyo,
* Corresponding author. Tel.: þ420 533435238; fax: þ420 533435313. E-mail addresses: [email protected], [email protected] (V. Antonı´n). 1340-3540/$ e see front matter ª 2013 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.myc.2013.07.003
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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m y c o s c i e n c e x x x ( 2 0 1 3 ) 1 e9
Japan). For lamellae, L means number of entire lamellae and l means number of lamellulae tiers between each pair of entire lamellae. For basidiospores, the factors E (quotient of length and width in any one spore) and Q (mean of E-values) are used. Color abbreviations follow Kornerup and Wanscher (1983), herbarium abbreviations Thiers (2013), and the authors of fungal names the Authors of Fungal Names page (http://www. indexfungorum.org/AuthorsOfFungalNames.htm).
2.2.
Molecular phylogeny
Molecular analyses such as DNA extraction, PCR amplification of ITS and LSU rRNA genes and the sequencing were performed according to the methods by Antonı´n et al. (2010a). Phylogenetic analyses were generated in MrBayes version 3.1.2 (Ronquist and Huelsenbeck 2003), applying a GTR þ I þ G substitution model with 1 106 generations, saving a tree every 10,000th generation, with the first 1000 trees discarded as burn-in for ITS and LSU datasets. The two analyses were performed atleast four times and were combined in a 50% majority rule consensus tree to obtain estimates for the posterior probabilities (PPs) by Antonı´n et al. (2010b). ITS and LSU dataset of M. siccus were chosen as an outgroup for rooting purpose based on a study by Tan et al. (2009) and Wannathes et al. (2009). The alignment datasets were deposited in TreeBASE (http://www.treebase.org) under ID S14462. For the list of specimens studied, including GenBank accession numbers, see Table 1.
3.
Results and discussion
3.1.
Taxonomy
Marasmius subsect. Marasmius Synonyms: Marasmius subsect. Pararotulae (Singer) Singer (type species: M. pararotula Singer). Marasmius subsect. Penicillati Singer (type species: M. graminum (Lib.) Berk.).
Type species: Marasmius rotula (Scop.: Fr.) Fr. Species are characterized in having pileipellis consisting of broom cells of the Rotalis-type. Marasmius wisteriae Antonı´n, R. Ryoo & H.D. Shin, sp. nov. Figs. 1A, 2. MycoBank no.: MB 803299. Pileus brownish orange to whitish, central dot dark brown or black. Basidiospores 11.5e13.5 2.5e3.2 mm, clavatefusoid, fusoid. Cheilocystidia and pileipellis cells of the Rotalis-type. Holotype: Republic of Korea, Chuncheon, Kangwon University experimental forest, 15. VI. 2009 leg. V. Antonı´n and R. Ryoo (BRNM 718761). rRNA gene sequences ex-holotype: JN003839 (ITS), JN003844 (LSU). Etymology: wisteriae, found on leaves of Wisteria floribunda. Pileus up to 2.5 mm broad, convex-conical with central umbilicus with dark brown or black dot, striateesulcate with crenulate, inflexed to straight margin, finely tomentose, pale brownish orange (paler than 6C4) to whitish. Lamellae distant, L ¼ 8e12, l ¼ 0, collariate, pale cream with concolorous, finely pubescent edge. Stipe up to 30 mm long, glabrous, lustrous, concolorous with lamellae and translucent at apex, brown, black-brown towards base. Basidiospores 11.5e13.5 (e14) 2.5e3.2 mm, 12.8 2.9 mm on average, E ¼ 3.8e5.2, Q ¼ 4.45, clavate-fusoid, fusoid, narrowly lacrimoid. Basidia (only one found) 21 8.0 mm, 4-spored, clavate. Basidioles 15e26 (2.5e) 4.0e8.0 mm, clavate, fusoid. Cheilocystidia similar to pileipellis broom cells, 12e20 7.0e13 mm, (broadly) clavate to pyriform, subvesiculose, diverticulate, thin-walled. Trama hyphae cylindrical, thin-walled, up to 10 mm wide. Pileipellis a hymeniderm composed of 14e25 10e15 mm, clavate, pyriform to subvesiculose cells of the Rotalis-type, thin-walled; diverticula digitate, thick-walled, obtuse. Stipitipellis a cutis of cylindrical, slightly thick-walled, smooth, up to 5.0 (e6.0) mm wide hyphae
Table 1 e Data on Marasmius specimens used to this study. Species
Marasmius bulliardii agg. M. bulliardii M. rotalis agg. M. rotalis agg. M. rotalis agg. M. rotula M. rotula M. rotula M. ruforotula M. ruforotula M. siccus M. siccus M. tubulatus M. wisteriae M. wisteriae
Locality
Korea Hungary Cameroon Korea Korea Germany Denmark USA Korea Korea Korea Russia Korea Korea Korea
Specimen ID
Ryoo KG 154, ¼BRNM 724476 Antonı´n 06.160, ¼BRNM 705006 Antonı´n Cm 01.107, ¼BRNM 666156 Ryoo KG 132, ¼BRNM 724479 Ryoo KG 232, ¼BRNM 724477 GLM 45962 Landvik NN005958 PBM 2563 Antonı´n 07.158, ¼BRNM 714674 Ryoo KG 107, ¼BRNM 714676 Ryoo KG 028, ¼BRNM 714680 Landvik NN050245 Antonı´n 07.160, ¼BRNM 714675 Antonı´n 09.102, ¼BRNM 718761 Ryoo KG 223, ¼BRNM 724478
GenBank accession number ITS
LSU
No data JN540056 KC415763 JN003837 KC415765 No data JN943598 DQ182506 FJ936150 FJ936152 FJ904985 JN943596 FJ936151 JN003839 JN003838
JN003846 JN540057 KC415764 JN003843 No data AY207238 JN941146 No data FJ917612 FJ917614 FJ904980 JN941142 FJ917613 JN003844 No data
Species names in bold type were retrieved from GenBank. The others were sequenced in this study. The herbarium abbreviations follow Thiers (2013).
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 1 e Basidiocarps of Marasmius species from South Korea. A: M. wisteriae (Ryoo KG 223). B: M. tubulatus (Antonı´n 07.160). C: M. rotalis (Ryoo KG 232). D: M. bulliardii (Ryoo KG 154). E: M. ruforotula (Antonı´n 07.158).
with brown walls in KOH. Clamps present. Chemical reactions: Trama, stipe medulla and stipitipellis hyphae non-dextrinoid or slightly dextrinoid, other structures non-dextrinoid. Habitat: On leaves of Wisteria floribunda, and on dead fallen leaves. Specimens examined: Republic of Korea, Chuncheon, Kangwon University experimental forest, alt. c. 1080 m, coord. 37 480 48.6900 N, 127 510 04.1400 E, 15 July 2009 leg. V. Antonı´n (09.102) and R. Ryoo (holotype, BRNM 718761); Seoul, Bukhansan National Park, in the vicinity of Jeongreung stream, 13 July 2008 leg. R. Ryoo (KG 223) (BRNM 724478). ? Japan, Miyazaki, Miyazaki College of Agri-culture, 29 June 1939 leg. Y. Arataka (K(M) 171077, as M. rotula). Marasmius wisteriae is especially characterized by a pale colored pileus with a dark central dot, distant lamellae, and large, clavate-fusoid, fusoid, narrowly lacrimoid basidiospores. Such combination of characters is not published in any described species. Marasmius pandoanus Singer has a white pileus and larger basidiospores (14.5e16.5 3.7e4.6 mm); M. oaxacanus Singer has also a white pileus, more close lamellae (L ¼ 10e16) and larger basidiospores (10e14 3e4 mm); M. arimanus Dennis differs by colored lamella edge and larger basidiospores (11e14 3.5e4 mm) (Singer 1976). A microscopically very similar specimen from Japan has been found in the K(M) herbarium. Its macroscopic description is not available, therefore, the exact identification is not possible (mentioned with a question mark in the list above).
Marasmius tubulatus Petch, Trans. Br. mycol. Soc. 31: 42, 1948. Figs. 1B, 3. Pileus up to 3 mm broad, convex-conical with obtuse center, then convex or almost applanate with slightly depressed center and inflexed margin, finely tomentose, entirely striateesulcate with crenulate margin, uniformly light brown to brownish orange (6C4). Lamellae distant, L ¼ 6e7, whitish, then dirty pale cream, with concolorous, later granulose-colored (pileus color) edge. Stipe up to 10 mm long, glabrous, brown (6E5e6) to black-brown with paler apex, originating from substrate. Rhizomorphs smooth, black-brown. Basidiospores (collapsed, only a few found) (7.0e) 8.0e10 3.75e4.5 mm, 9.0 4.0 mm on average, E ¼ 1.9e2.4, Q ¼ 2.2, ellipsoid-fusoid or subfusoid. Basidioles 12e20 4.0e9.0 mm, clavate, fusoid. Cheilocystidia of the Rotalis-type, 15e19 8.0e11 mm, (broadly) clavate to pyriform, thin-walled, with pale (yellow) gray-brown walls in KOH. Pileipellis a hymeniderm composed of 12e25 10e14 mm, clavate to pyriform cells of the Rotalistype, thin-walled; diverticula up to 3.0 1.0 mm, digitate, often irregular; thick-walled parts (yellow) gray-brown in KOH. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps present. Chemical reactions: Trama, stipitipellis and stipe medulla hyphae (slightly) dextrinoid, other structures nondextrinoid. Habitat: On dead twigs of Salix sp. Specimen examined: Republic of Korea, Namyangju, Korea University experimental farm, alt. c. 140 m, 37 340 5200 N, 127
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 2 e Marasmius wisteriae (BRNM 718761). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
140 4000 E, 30 July 2007 leg. V. Antonı´n (07.160), H.D. Shin, R. Ryoo and M.J. Park (BRNM 714675). Our collection of M. tubulatus is characterized by a small uniformly light brown to brownish orange pileus, very distant lamellae (L ¼ 6e7) with a concolorous or granulose-colored edge, a short stipe not originating from rhizomorphs, moderately large basidiospores, and pileipellis cells of the Rotalis-type. Marasmius tubulatus has been mentioned by various authors in various senses with different character combinations. A description by Manimohan and Leelawathy (1989) is the closest one to our fungus. Description by Tan et al. (2009) differs by slightly closer lamellae (L ¼ 10e13) and larger cheilocystidia (12e27 7e18 (e22) mm).
Marasmius bulliardii agg. Que´l., Bull. Soc. bot. Fr. 24: 323, 1878. Fig. 1C. Pileus conical, then convex with depressed center and central dot or small papilla in it, sulcate, ochraceous brown, whitish to pale brownish at center. Lamellae distant, L ¼ 15e17, pale cream, then darker, with concolorous edge. Stipe glabrous, dark brown to black-brown. Rhizomorphs probably absent. (Description based on dry basidiocarps and photographs.) Basidiospore (only one found) 8.5 5.5 mm, E ¼ 1.55, ellipsoid. Hymenium damaged. Pileipellis a hymeniderm of 20e35 (e45) 17e25 mm, broadly clavate or vesiculose broom cells of the Rotalis-type, thin- to slightly thick-walled; projections up to 3.0 1.5 mm; thickwalled parts of cells and projections brown in KOH.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 3 e Marasmius tubulatus (BRNM 714675). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Stipitipellis a cutis of cylindrical, up to 4.0 mm wide hyphae with dark brown walls in KOH. Clamps present. Chemical reactions: Stipitipellis and stipe medulla hyphae (slightly) dextrinoid at apex, other structures non-dextrinoid. Habitat: On dead leaves. Specimen examined: Republic of Korea, Deogyusan National Park, Cheon-yeon falls, 24 Aug. 2007 leg. R. Ryoo (KG 154) (BRNM 724476). The characters agree with M. bulliardii, and LSU sequences are also similar to the European taxon from Hungary (Table 1, Fig. 7). Microscopically, it differs only by larger pileipellis cells. However, material is very poor, a macroscopic description of fresh basidiocarps is absent, and, moreover, hymenium is damaged. Therefore, it is necessary to re-collect it to settle its taxonomic status. Marasmius wettsteinii Sacc. & P. Syd. published by Cho and Yoo (1999) from the Republic of Korea may represent our collection. However, the authors mentioned very small basidiospores (5e6 3e4 mm).
Marasmius rotalis agg. Berk. & Broome, J. Linn. Soc., Bot. 14: 40, 1873. Figs. 1D, 4. Pileus conical with depressed center and dark central dot, sulcate, finely tomentose, pale greyish with slightly paler margin. Lamellae distant, L ¼ 13e16, white, with concolorous edge. Stipe glabrous, black-brown. Rhizomorphs (probably) absent. (Description based on dry basidiocarps and photographs.) Basidiospores 7.5e9.0 (4.25e) 4.5e5.0 mm, 8.4 4.7 mm on average, E ¼ 1.6e2.0, Q ¼ 1.8, ellipsoid. Basidioles 15e29 4.0e7.0 mm, clavate, cylindrical or fusoid. Cheilocystidia 11e24 7.0e15 mm, of the Rotalistype, clavate, (sub)vesiculose, hyaline. Pileipellis a hymeniderm of (12e) 15e25 (e30) 8.0e20 mm, pyriform, broadly clavate or subvesiculose broom cells of the Rotalis-type, thin- to slightly thick-walled at apex. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps present. Chemical reactions: Stipitipellis and stipe medulla hyphae non-dextrinoid or weakly dextrinoid, other structures non-dextrinoid.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 4 e Marasmius rotalis (BRNM 724477). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Habitat: On dead leaves (e.g., Quercus). Specimen examined: Republic of Korea, Pyeongchang, Chungtaesan Resort Forest, 17 July 2008, leg. R. Ryoo (KG 232) (BRNM 724477). Our collection agrees with descriptions of M. rotalis (e.g., Antonı´n 2007), however, Singer (1976) described it without a dark central dot. Desjardin and Horak (1997) mentioned only 7.5e9 3e3.5 mm large basidiospores. The type specimen of M. rotalis (Sri Lanka, Peradeniya, K(M)92650!) differs by having slightly narrower basidiospores (7.0e10 3.5e 4.5 mm) and smaller pileipellis broom cells ((12e) 15e17 5.0e12 mm). Another very similar South-Korean collection (Chuncheon, Dongsan-myeon, Bongmyong-ri, 20 Aug. 2007 leg. R. Ryoo KG 132, BRNM 724479) forms, according to LSU and ITS, the same tree with the collection of M. rotalis from Cameroon (prov. Sud, Dja Biosphere Reserve, Nkoelgnegue
near Ekom, ca. 37 km to E of Somalomo, 12 Apr. 2001 leg. V. Antonı´n, BRNM 666156). However, the Korean collection is too poor for detailed microscopic studies. Both above mentioned collections are close in comparison of LSU sequences with two sequences of M. rotula taken from GenBank. However, both species groups distinctly differ by their macroscopic characters. Marasmius rotalis species complex is very complicated in East and South-East Asia (cf., Tan et al. 2009; Wannathes et al. 2009). Marasmius subsect. Sicciformes Antonı´n, Acta Mus. Moraviae, Sci. nat., 76: 145. 1991. Type species: Marasmius curreyi Berk. & Broome Species are characterized in having pileipellis consisting of broom cells of the Siccus-type. Marasmius ruforotula Singer, Sydowia 2: 34, 1948. Figs. 1E, 5.
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Fig. 5 e Marasmius ruforotula (BRNM 714676). A: Pileipellis cells. B: Basidiospores. C: Cheilocystidia. Bar 20 mm.
Pileus 1e7 mm broad, convex-hemispherical or conical, then (flat-)convex, with depressed center and small papilla, sulcate, finely tomentose, dark brown or reddish brown (7e8BeD6e8), darker reddish brown (8E8 to 9CeDeE8) at center, paler (7e8C5e6) when old, paler in striae to less distinctly striped, almost cream-whitish at very margin and in oldest (revived?) basidiocarps. Lamellae distant, L ¼ 9e11, slightly intervenose when old, pale cream, later yellowish white (3e4A2), with colored (pileus color) edge. Stipe up to 25 mm long, glabrous, dark brown to black-brown (7EeF6e7). Rhizomorphs present. Basidiospores 7.5e8.5 (e9.5) 4.0e5.0 mm, 8.4 4.5 mm on average, E ¼ 1.5e2.1, Q ¼ 1.8e1.9, ellipsoid, ellipsoid-fusoid. Basidia 23e30 7.0e10.5 mm, 4-spored, clavate. Cheilocystidia of the Siccus-type, 11e19 8.0e12 mm, (broadly) clavate to pyriform, thin-walled with thick-walled apex, mixed with scattered smooth, clavate, thin-walled, regular to irregular cells with brown walls in KOH. Pileipellis a hymeniderm of (9.0e) 12e18 7.0e11 mm, clavate, pyriform or vesiculose, often irregular broom cells of the Siccus-type, thin-walled with slightly thick-walled apex; diverticula 4e20, up to 6.0 1.0 mm, cylindrical to conical; mixed with irregular, thickwalled cells (almost) without projections; thick-walled parts (reddish) brown in KOH. Stipitipellis a cutis of cylindrical, up to 5.0 mm wide hyphae with brown walls in KOH. Clamps
present. Chemical reactions: Stipitipellis, pileus context and stipe medulla hyphae (slightly) dextrinoid, other structures non-dextrinoid. Habitat: On dead twigs of a broadleaved tree (Salix sp.?) and on dead stems of grass and Juncus sp. Specimens examined: Republic of Korea, Namyangju, Korea University experimental farm, alt. c. 140 m, 37 340 5200 N, 127 140 4000 E, 30 July 2007 leg. V. Antonı´n (07.158), H.D. Shin, R. Ryoo and M.J. Park (BRNM 714674); Yangpyeong, Jungmisan National Resort Forest, 37 340 4600 N, 127 270 0700 E, alt. c. 395 m, 1 July 2008 leg. V. Antonı´n (08.47) and R. Ryoo (BRNM 718685); Guri, Dongguneung (East Nine Tombs), alt. 30 m, 37 370 0300 N, 127 080 0100 E, 16 Aug. 2007 leg. R. Ryoo (KG 107) (BRNM 714676). In literature M. ruforotula has been mentioned in the several senses (e.g., Singer 1976; Pegler 1983; Desjardin and Horak 1997). They especially differ each other by the basidiospore size. Our collections fit to the description by Desjardin et al. (2000) from Indonesia. The Yangpyeong collection (BRNM 718685) differs by the presence of a dark central pileus dot, slightly more close lamellae (L ¼ 11e13), an up to 40 mm long stipe, and the presence of smooth cells among typically developed cheilocystidia.
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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Fig. 6 e Phylogenetic tree of Marasmius sect. Marasmius inferred from Bayesian analysis of the ITS region (ITS1-5.8S-ITS2), showing mean branch lengths of a 50% majority rule consensus tree, obtained from an MCMC analysis of 106 generations. Species names in bold type were retrieved from GenBank.
Fig. 7 e Phylogenetic tree of Marasmius sect. Marasmius inferred from Bayesian analysis based on the LSU rRNA gene sequences, showing mean branch lengths of a 50% majority rule consensus tree, obtained from an MCMC analysis of 106 generations. Species names in bold type were retrieved from GenBank.
3.2. Key to the species in Marasmius sect. Marasmius found in the Republic of Korea 1 Cheilocystidia and pileipellis broom cells in the form of Siccus-type (subsect. Sicciformes) ........................................................................................... M. ruforotula 1 Cheilocystidia and pileipellis broom cells in the form of Rotalis-type (subsect. Marasmius) ................................................................................................................. 2 2 Basidiospores 12–13.5(–14) × 2.5–3.25 µm, fusoid, narrowly lacrimoid M. wisteriae 2 Basidiospores 7.0–10 × 3.75–5.0 µm, ellipsoid to ellipsoid-fusoid ........................... 3 3 Pileus ochraceous brown except for whitish centre; lamellae moderately distant (L = 15–17); basidiospore 8.5 × 5.5 µm, E = 1.55; pileipellis cells 20–35(–45) × 17–25 µm; on dead leaves ................................................................................... M. bulliardii 3 Pileus pale greyish or light brown to brownish orange; basidiospores 2.5–4.5 µm broad, Q = 1.8–4.5 ....................................................................................................... 4 4 Pileus light brown to brownish orange; lamellae distant (L = 6–7); basidiospores (7.0–)8.0–10 × 3.75–4.5 µm, Q = 2.2; on twigs ...................................... M. tubulatus 4 Pileus pale greyish; lamellae more close (L = 13–16); basidiospores 7.5–9.0 × 4.5– 5.0 µm, Q = 1.8; on dead leaves .................................................................... M. rotalis
3.3.
Phylogenetic analysis
Phylogenetic trees of the genus Marasmius sect. Marasmius were inferred from MCMC analyses based on ITS and LSU rRNA gene
sequences obtained in this study and retrieved from GenBank (Figs. 6, 7). For ITS and LSU rRNA gene dataset, 13 and 12 sequences were aligned and trimmed to 514 and 897 base pairs at each other. The analyzed sequences are divided in two groups following subsections in this topology. The one group consisted of subsect. Marasmius and the other included M. ruforotula (subsect. Sicciformes). The results are concordant with the delimitations of five species based on macro- and micromorphological characteristics. Marasmius wisteriae forms a distinct clade. Marasmius rotalis formed the same subclade with sequences of M. rotula from GenBank, type species of the genus Marasmius in both datasets (see remarks in M. rotalis). Although two specimens of this species were collected in Korea, they represented the distinct phylogenetic position in ITS dataset. One of them, BRNM 724479, formed a grouping with Cameroon in LSU as well as ITS of phylogenetic tree. We excluded samples BRNM 724477 and 724478, from the phylogenetic analysis of LSU rRNA genes owing to its amplifications of multiple fragments. Because a very small quantity of DNA was extracted from M. bulliardii, BRNM 724476, we couldn’t obtain ITS sequences of this isolate and be included in a phylogeny tree. LSU sequences of
Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003
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this isolate, BRNM 724476, however, were similar to those of M. bulliardii collected from Hungary. They were different 3 out of 918 nucleotide characters. Phylogenetic analysis of ITS and LSU rRNA genes for Marasmius sect. Marasmius suggested the independent taxonomic position of morphologically similar species, although they were limited by the absence of sequences of several isolate.
4.
Conclusions
Studies of South-Korean Marasmius sect. Marasmius specimens confirmed the presence of five taxa till now. Macro- and microscopical characters of some species of this section (see e.g., discussion on M. bulliardii and M. rotalis in this study) are very similar to almost identical, and it is very difficult to obtain exact taxonomic results without using DNA studies. Moreover, it seems that many Marasmius species have their morphologically very similar vicariant taxon in other geographical areas (Antonı´n unpublished studies, Petersen pers. comm.).
Acknowledgements The collecting trip to the Republic of Korea and the studies of the material by the first author were supported by the Czech Science Foundation (No. 206/07/J003), and due the support provided to the Moravian Museum by the Ministry of Culture of the Czech Republic as part of its long-term conceptual development programme for research institutions (ref. MK000094862). The other authors were supported by the Korea Research Foundation Grant funded by the Korean Government (KRF-2006-F00001) and by project (FP 0801-2009) of Korea Forest Research Institute.
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Please cite this article in press as: Antonı´n V, et al., Marasmioid and gymnopoid fungi of the Republic of Korea. 6. Marasmius sect. Marasmius, Mycoscience (2013), http://dx.doi.org/10.1016/j.myc.2013.07.003