MYCOBIOLOGY 2020, VOL. 48, NO. 4, 252–262 https://doi.org/10.1080/12298093.2020.1769541
RESEARCH ARTICLE Marasmioid and Gymnopoid Fungi of the Republic of Korea. 8. Gymnopus Section Levipedes
Rhim Ryooa, Vladim ır Anton ınb and Kang-Hyeon Kaa aDepartment of Forest Bioresources, National Institute of Forest Science, Suwon, Republic of Korea; bDepartment of Botany, Moravian Museum, Brno, Czech Republic
ABSTRACT ARTICLE HISTORY Collections of Gymnopus sect. Levipedes from the Republic of Korea have been studied. Two Received 21 July 2019 new species, Gymnopus dryophiloides and G. brunneodiscus, are described based on their Revised 6 May 2020 macro- and micromorphological and phylogenetic characteristics. Three other species, Accepted 11 May 2020 Gymnopus referred to as spp. 1, 2, and 3, are distinguished as separate taxa without formal KEYWORDS descriptions. Taxonomic and phylogenetic positions of all taxa have been inferred and con- Omphalotaceae; molecular firmed by analyses of ITS and LSU sequence data. Their detailed descriptions, illustrations systematics; new taxa; and an identification key are provided. ITS; LSU
1. Introduction 2. Material and Methods The genus Gymnopus (Pers.) Roussel belongs to the 2.1. Morphology family Omphalotaceae, and is, according to Anton ın Macroscopic descriptions of collected specimens are and Noordeloos [1], divided into five sections. based on fresh basidiomata. Color abbreviations fol- Species of sect. Levipedes (Fr.) Halling are character- low Kornerup and Wanscher [13]. The authors of ized by a pileipellis composed of inflated, mostly fungal names are cited according to the irregular, lobed or coralloid elements (Dryophila- International Plant Names Index Authors website structure). Other Gymnopus sections have a pileipel- (http://www.ipni.org/ipni/authorsearchpage.do). lis in the form of an (ixo)cutis or an irregular tri- Microscopic features were studied using dried choderm [1,2]. There are about 45 species described material mounted in H2O, approximately 5% KOH, worldwide and their distribution is very broad Melzer’s reagent, and Congo Red (for recipes, see [1–8]. However, information from many parts of [14]) using an Olympus BX-50 light microscope the world (e.g., Africa, South America) is missing or (Tokyo, Japan) at 1000 magnification. For lamel- very incomplete. lae, L refers to the number of entire lamellae, while None of the species belonging to Gymnopus sect. l refers to the number of lamellulae tiers between Levipedes had been recorded in the Republic of each pair of entire lamellae. For basidiospores, the Korea until 2011 [9]. Jang et al. [10] published their factor E indicates the quotient of the length and own Korean sequence called G. dryophilus in the width in any one basidiospore and Q is the mean of the E-values; the basidiospore values are based on phylogeny tree (but see our results, Table 1, 20 measurements in each collection. Specimens are Figure 1). preserved in the herbarium of the Moravian Specimens of Gymnopus sect. Levipedes collected Museum, Brno, Czech Republic (BRNM). Cultures from various localities throughout the Republic of are housed in the herbarium of the National Korea since 2007 are evaluated in this study. This Institute of Forest Science (NIFoS), Republic paper is one of a series of papers describing the of Korea. diversity of marasmioid, marasmielloid, and gymno- poid fungi in the Republic of Korea, supported by the National Institute of Forest Science. Two papers 2.2. Phylogenetic analyses dealing with sections Androsacei and Impudicae of DNA extraction was performed by means of the the genus Gymnopus have already been pub- modified method by Lee and Taylor [15]. The PCR lished [11,12]. primer used ITS1F, ITS1, ITS4B and ITS4 for the
CONTACT Vladim ır Anton ın [email protected] ß 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. MYCOBIOLOGY 253
Table 1. The list on the GenBank accession number of nrITS and nrLSU sequences used for this study. GenBank accession Number Scientific name Herbarium Number Locality ITS LSU Gymnopus alpinus Duke15 Sweden DQ480101 – Gymnopus alpinus Duke14 U.K., Scotland DQ480102 – Gymnopus alpinus TENN 55,834 U.K., Scotland DQ480114 – Gymnopus alpinus CB 16251 Latvia JX536168 – Gymnopus aquosus TENN 57958 Germany AY256691 – Gymnopus aquosus TENN 55,883 U.K., Scotland DQ450003 – Gymnopus aquosus BRNM 710027 Czech Republic JX536170 – Gymnopus aquosus BRNM 670755 Czech Republic JX536171 – Gymnopus aquosus BRNM 695556 Czech Republic JX536173 – Gymnopus aurantiipes SFSU-AWW118 Java AY263432 – Gymnopus bicolor SFSU-AWW116 Java AY263423 AY639411 Gymnopus brunneodiscus BRNM 808974 Korea MH589971 MH589989 Gymnopus brunneodiscus BRNM 808971 Korea MH589972 MH589992 Gymnopus brunneodiscus BRNM 714974 Korea MH589973 MH589988 Gymnopus brunneodiscus BRNM 808976 Korea MH589974 MH589990 Gymnopus brunneodiscus BRNM 808975 Korea MH589975 MH589991 Gymnopus confluens TENN F-059603 Russia KP710300 – Gymnopus confluens BRNM 734005 Czech Republic JX536124 – Gymnopus confluens TENN F-059603 Sweden – KP710313 Gymnopus confluens GLM 45930 Germany – AY207164 Gymnopus dryophilus TENN 58257 Russia DQ449963 – Gymnopus dryophilus TENN 52412 USA, ID DQ449964 – Gymnopus dryophilus TENN 51438 Canada, NS DQ449966 – Gymnopus dryophilus Duke29 DQ480098 – Gymnopus dryophilus BRNM 695586 Czech Republic JX536143 – Gymnopus dryophilus BRNM 712600 Czech Republic JX536158 – Gymnopus dryophilus TENN 57012 – AY640619 Gymnopus dryophilus ZRL201566 China – KY418871 Gymnopus dryophilus FO 21603 Germany – AF291305 Gymnopus dryophilus TENN 57012 – NG027632 Gymnopus dryophilus var. lanipes (isoneotype) BRNM 670686 Spain JX536137 – Gymnopus dryophilus (isoneotype) DUKE 193411 Sweden JX536153 – Gymnopus dryophiloides KUC20140627-37 Korea KX513744 – Gymnopus dryophiloides BRNM 781446 (specimen) Korea MH589959 – Gymnopus dryophiloides BRNM 781446 (culture) Korea MH589961 Gymnopus dryophiloides BRNM 781444 Korea MH589960 – Gymnopus dryophiloides BRNM 781448 Korea MH589962 MH589980 Gymnopus dryophiloides BRNM 781450 Korea MH589963 MH589983 Gymnopus dryophiloides BRNM 718679 Korea MH589964 - Gymnopus dryophiloides BRNM 781451 Korea MH589965 MH589981 Gymnopus dryophiloides BRNM 781453 Korea MH589966 MH589977 Gymnopus dryophiloides BRNM 781447 Korea MH589967 MH589985 Gymnopus dryophiloides BRNM 781452 Korea MH589970 MH589984 Gymnopus dryophiloides BRNM 781449 Korea – MH589978 Gymnopus dryophiloides BRNM 781454 Korea – MH589979 Gymnopus dryophiloides BRNM 781456 Korea – MH589982 Gymnopus earleae TENN 59457 USA, OR AY256694 – Gymnopus erythropus SAV XI 2002 Slovakia DQ449996 – Gymnopus erythropus BRNM 705224 Czech Republic JX536131 – Gymnopus erythropus BRNM 664995 Czech Republic JX536133 – Gymnopus erythropus BRNM 693553 Switzerland JX536135 – Gymnopus erythropus GLM 45932 – DQ071804 Gymnopus fagiphilus BRNM 712422 Czech Republic JX536125 – Gymnopus fagiphilus BRNM 712407 Czech Republic JX536126 – Gymnopus fagiphilus BRNM 691489 Czech Republic JX536128 – Gymnopus hybridus Dvo r ak 138/02, BRNU Czech Republic JX536175 – Gymnopus hybridus Dvo r ak 393/07, BRNU Czech Republic JX536176 – Gymnopus hybridus BRNM 695773 Italy JX536177 – Gymnopus indoctoides AWW125 AY263424 AY639419 Gymnopus inusitatus SCM B-4065 Spain JN247551 JN247555 Gymnopus inusitatus SCM B-4057 Spain JN247552 JN247556 Gymnopus inusitatus SCM B-4058 Spain JN247553 JN247557 Gymnopus inusitatus var. cystidiatus BRNM 737257 Hungary – JN247554 Gymnopus junquilleus TENN 59532 USA, TN AY256693 – Gymnopus macropus TENN 58090 Costa Rica AF505788 – Gymnopus macropus TENN 56636 Costa Rica DQ449978 – Gymnopus macropus TENN 58619 Costa Rica DQ449979 – Gymnopus nubicola NY REH8290 Costa Rica AF505781 – Gymnopus ocior BRNM 728565 Slovakia JX536160 – Gymnopus ocior BRNM 737693 Norway JX536164 – Gymnopus ocior BRNM 699795 Czech Republic JX536166 – Gymnopus sepiiconicus AWW117 Java AY263448 – Gymnopus sepiiconicus AWW126 – AY639427 Gymnopus subsulphureus TENN 56718 USA, NC AY256692 – Gymnopus subsulphureus TENN 56321 USA, NC DQ449972 – Gymnopus spongiosus Duke229 DQ480093 – Gymnopus spongiosus TENN 59640 USA, Tennessee DQ480113 – Gymnopus vitellinipes AWW115 AY263453 – Gymnopus sp.1 BRNM 808978 Korea MH589969 MH589986 Gymnopus sp.2 BRNM 718669 Korea MH589976 MH589993 Gymnopus sp.3 BRNM 718701 Korea MH589968 MH589987 Entries in bold type were newly sequenced by the authors. 254 R. RYOO ET AL.
Figure 1. Phylogenetic tree for Gymnopus sect. Levipedes based on the nrITS region (ITS1, 5.8S and ITS2) inferred from Bayesian analysis. Specimen names and their GenBank accession numbers in bold letters were newly obtained for this study. nuclear ribosomal internal transcribed spacer from GenBank [4,5,18]. Eighteen sequences of the (nrITS) region and selected LR0R, and LR7 for the nrITS dataset and 17 sequences of the nrLSU dataset nuclear large subunit ribosomal DNA (nrLSU) were formed for this study. Gymnopus confluens region according to Gardes and Bruns [16]. PCR belonging to section Vestipedes was selected as an cycling condition and DNA sequencing was carried outgroup [18]. GenBank accession numbers of all out according to the protocol described by Anton ın sequences for the phylogenetic trees are listed in et al. [12,17]. For the nrITS region, PCR was per- Table 1. Both the nrITS and nrLSU datasets were formed with a 30 s denaturation at 94 C, a 30 s aligned with Mega 6.06 [19] and Clustal X2 [20]. annealing at 56 C and a 1 min extension at 72 C Each phylogenetic analysis was performed by and was run with 35 cycles in the first denaturation MrBayes 3.1.2. [21]. In the nrITS and nrLSU data- and the last extension. For the nrLSU region, PCR sets, the ends of them were trimmed to 714 and 882 was carried out with a 30 s denaturation at 94 C, a characters including 492 and 796 constant, and 188 45 s annealing at 45 C and a 90 s extension at 72 C and 46 variable sites, respectively. Both Markov and then 35 cycles were run with the first denatur- chains were run for 2,000,000 generations with a ation and the last extension. Forty nrITS sequences sample taken every 100th generation, discarding a of 19 species and 12 nrLSU sequences of eight spe- burn-in of 1,000 generations. The general time cies belonging to section Levipedes were retrieved reversible (GTR) was used with gamma-distributed MYCOBIOLOGY 255 substitution for a given data set. The Bayesian of 714 nucleotide characters of the nrITS dataset branch supports were assigned as posterior probabil- from G. ocior (Pers.) Anton ın & Noordel. They have ities (PP) according to the 50% majority-rule of the a sequence divergence of about 5.9%. The independ- consensus trees. The PP values showed to be higher ent taxonomic positions of Gymnopus spp. 1 and 2 than 0.95, including 95% confidence intervals (CI). are supported by the nrITS and nrLSU sequences. Gymnopus sp. 1 and five sequences of G. aquosus (Bull.: Fr.) Anton ın & Noordel. form one clade and 3. Results differ in 7 nucleotide sites, although the two species 3.1. Phylogenetic analyses are morphologically different. In the nrITS dataset, Gymnopus sp. 3 forms a group with G. erythropus The two newly described species, Gymnopus dryo- (Pers.: Fr.) Anton ın, Halling & Noordel., G. earleae philoides and G. brunneodiscus, and Gymnopus spp. Murrill, G. fagiphilus (Velen.) Anton ın, Halling & 1, 2 and 3, are distinguished from morphologically Noordel., G. hybridus (Kuhner€ & Romagn.) Anton ın similar species by phylogenetic analyses of nrITS & Noordel., G. inusitatus (Vila & Llimona) Vila & and nrLSU (Figures 1 and 2). In two phylogenetic Llimona, G. spongiosus (Berk. & M.A. Curtis) trees, taxa of the Gymnopus dryophilus complex col- Halling, and G. vitellinipes A.W. Wilson, Desjardin lected in Korea showed a monophyletic clade (95% & E. Horak. This species forms also a significantly and 98%), and formed one other clade close to distant branch with G. earleae according to 20 base European and American taxa with high supporting substitutions with 714 base pairs of nrITS sequences values (100% and 100%). This clade is described as and has a sequence divergence of 2.8%. a new species, G. dryophiloides. In the nrITS region, this species have a sequence divergence of 1.3% and differ in 9 out of 714 nucleotide sequences from 3.2. Taxonomy typical G. dryophilus (isolate; AFTOL_ID 559, 3.2.1. Species descriptions GenBank; DQ241781). In the nrLSU region, this Gymnopus dryophiloides Anton ın, R. Ryoo & K.H. new species has different sequences within 3 bp Ka, sp. nov. among 822 bp in several isolations from other local- MycoBank MB 828254 ities (China, Germany, USA). In the monoclade Diagnosis: Gymnopus dryophiloides differs from including holotype G. dryophiloides, the nrITS G. dryophilus by a more distinct orange tinge of the sequence (MH589967) of the G. dryophiloides holo- stipe, slightly smaller (shorter) basidiospores, shorter type differs the only 1 bp from KX513744, 2 bp with cheilocystidia with a larger variability in shape, and MH589958, and the nrLSU sequences have the same forms an independent monoclade in the ITS and base pair. Gymnopus brunneodiscus differs in 42 out LSU dataset. Gymnopus dryophiloides is
Figure 2. Phylogeny of Gymnopus sect. Levipedes based on the nrLSU (nuclear large subunit of ribosomal RNA gene) inferred from Bayesian analysis. Specimen names and their GenBank accession numbers in bold letters were newly sequenced for this study. 256 R. RYOO ET AL.
Figure 3. Basidiomata. (A) Gymnopus dryophiloides (holotype, BRNM 781447); (B) G. dryophiloides (BRNM 781452); (C) G. brun- neodiscus (holotype: BRNM 808975); (D) Gymnopus sp. 3 (BRNM 718701); (E) Gymnopus sp. 1 (BRNM 808978); (F) Gymnopus sp. 2 (BRNM 718669). characterized by a pale to light yellow, orange or Lamellae very close, L ¼ (35)50–60, l ¼ 3–4(6), watery ochraceous yellow pileus. It has yellowish emarginate and attached with small tooth, narrow, white lamellae, a pale to light yellow or orange stipe, whitish then yellowish white (3A2–3(4)), with a (4.0)4.5–6.0(6.5) 2.5–3.5(4.0) lm large basidio- concolorous, finely denticulate and pubescent edge. spores, and 17–38(50) 4.5–10(17) lm large, cla- Stipe 30–75 1.5–5 mm, cylindrical or laterally com- vate, (sub)cylindrical, fusoid, mostly lobate, irregular pressed (sometimes with groove), slightly broadened to coralloid cheilocystidia (Figures 3 and 4). at apex, slightly broader to subbulbose (up to Holotype: Republic of Korea. Mt. Kariawangsan, 10 mm) at base, bulba more distinct in young basi- Dae-hwa-myeon, Pyeonchang, 37 2803200N, 128 30ˊ diomata, glabrous or finely pruinose at apex, smooth 1100E, alt. c. 1060–1080 m, 21 August 2012, V. or finely longitudinally fibrillose, concolorous with Anton ın 12.138 & K.-H. Ka (BRNM 781447, desig- lamellae at apex, pale to light yellow or orange nated here). (4–5A3–6), sometimes darker, light orange or gray- Basidiomata single. Pileus 10–50 mm broad, con- ish orange (5A4, 6B6) in old basidiomata; with whit- vex to conical-convex with involute margin when ish to pale (pinkish) ochraceous basal rhizoids. young, then conical-convex to plano-convex with Context membranaceous, concolorous with surface less distinct to absent broad umbo and inflexed to of basidiomata, hollow in stipe, with slightly fungoid straight margin, often uplifted and undulate when smell and mild taste, sometimes with slightly old, hygrophanous, striate only at outer margin or acrid aftertaste. not translucently striate, smooth or slightly rugulose, Basidiospores (4.0)4.5–6.0(6.5) 2.5–3.5(4.0) mm, glabrous, pale to light yellow or orange (4–5A3–6, average ¼ 5.3 3.1 mm, E ¼ 1.50–2.40, Q ¼ 1.70–1.86, 6A–B5) or watery ochraceous yellow when moist, ellipsoid, ellipsoid-fusoid, pip-shaped, hyaline, thin- light yellow to yellowish white (3–4A2–3) when dry. walled. Basidia 17–23(25) 5.0–7.5 mm, 4-spored, MYCOBIOLOGY 257
Deogyusan National Park, Cheon-yeon falls, 24 August 2007, R. Ryoo KG 158 (BRNM 781446); Deogyusan National Park, Chilyeon falls, 23 August 2007, R. Ryoo KG 153 (BRNM 781446); Cheongju, Songnisan-maltijae Recreational Forest, 37 2903300N, 127 4604300E, alt. c. 230 m, 27 August 2015, V. Anton ın 15.106, K.-H. Ka, S.-K. Kim & J.-A. Kang (BRNM 781451); Chungju, Joryeongsan Natural Forest, 36 4804300N, 128 0204300E, alt. c. 495 m, 28 August 2015, V. Anton ın 15.117, K.-H. Ka, S.-K. Kim & J.-A. Kang (BRNM 781452); Chungju, Bonghwang Nature Forest, 37 0302000N, 127 4904600E, alt. c. 115 m, 29 August 2015, V. Anton ın 15.125, K.-H. Ka, S.-K. Kim & J.-A. Kang (BRNM 781453); Youngdong, Mt. Minjooji Recreational Forest, 36 0301200N, 127 4905000E, alt. c. 620 m, 26 August 2015, V. Anton ın 15.97, K.-H. Ka, S.-K. Kim & J.-A. Kang (BRNM 781450); Wonju, Mt. Chiaksan, 37 2502400N, 128 0401600E, alt. c. 290 m, 27 August 2015, V. Anton ın 15.75, K.-H. Ka, S.-J. Yeong & E.- Figure 4. Gymnopus dryophiloides (BRNM 781447). (A) chei- J. Wang (BRNM 781449); Jecheon, recreational for- 0 00 0 00 locystidia; (B) basidiospores; (C) pileipellis cells. Scale bar est, 37 08 42 N, 128 02 03 E, alt. c. 350 m, 21 ¼ 20 lm. August 2015, V. Anton ın 15.65, K.-H. Ka, S.-J. Yeong & E.-J. Wang (BRNM 781448); Muju, Mt. clavate, sometimes subcapitate. Basidioles Deogyu Recreational Forest, 35 5402600N, 12–25 3.0–8.0(9.0) mm, subcylindrical, clavate (and 127 4803900E, alt. c. 750 m, 30 August 2016, V. sometimes subcapitate), fusoid. Cheilocystidia Anton ın 16.130, K.-H. Ka & S.-H. Kim (BRNM 17–38(50) 4.5–10(17) mm, variable in shape, cla- 781454); ibid., 5 September 2016, V. Anton ın vate, (sub)cylindrical, fusoid, mostly lobate, irregular 16.155, R. Ryoo, K.H. Wang & Y.S. Jang (BRNM to coralloid, thin-walled. Pleurocystidia absent. 781456); Jangsu, Waryong National Recreational Trama hyphae cylindrical or subinflated, ± thin- Forest, 35 4004500N, 127 2803700E, alt. 570 m, 1 walled, branched, smooth or minutely incrusted, September 2016, V. Anton ın 16.145, K.-H. Ka & S.- 4.0–15 mm wide. Pileipellis composed of cells forming H. Kim (BRNM 781455); Gyeongnam Prov., a Dryophila-structure, up to 80 5.0–25 mm in size, Geochang-gun, Geumwonsan Natural Recreational cylindrical, fusoid or clavate, irregular, lobate, Forest, 35 4305500N, 127 4705200E, alt. 480 m, 11 July branched, subcoralloid, thin- to slightly thick-walled, 2017, V. Anton ın 17.25, K.-H. Ka, R. Ryoo & M.-H. smooth or incrusted, pale (dirty) yellow-brown in Jeon (BRNM 808970). KOH. Pileocystidia absent. Stipitipellis a cutis of Remarks: Gymnopus dryophiloides is character- cylindrical, parallelly arranged, slightly thick-walled, ized by having a small, light to grayish orange smooth, up to 6.0 mm wide hyphae. Caulocystidia pileus, a light yellow to light orange, at base subbul- absent, scattered cylindrical, thin-walled terminal bose stipe, small basidiospores, clavate, subcylindri- cells present. Clamp connections present in cal, fusoid, mostly lobate, irregular to coralloid all tissues. cheilocystidia, and a pileipellis of the Dryophila- Ecology: On detritus and litter of Abies holo- structure. A collection from Dongsan-myeon, phylla, Acer sp., Castanea crenata, Juglans man- Chuncheon (BRNM 781444) differs by smaller, dshurica, Larix campestris, Pinus densiflora, P. broadly clavate, fusoid, utriform, only more or less koraiensis, P. rigida, Quercus acutissima, Q. mongol- regular cheilocystidia, and a collection from ica, Q. serrata, Quercus sp. and Zelkova serrata. Deogyusan National Park, Chilyeon falls (BRNM Etymology: dryophiloides ¼ similar to 781446) also by mostly clavate, regular G. dryophilus. cheilocystidia. Additional Specimens Examined: Republic of Korea. Inje, Buk-myeon, Seoraksan National Park, Gymnopus brunneodiscus Anton ın, R. Ryoo & 0 00 0 00 38 09 51 N, 128 22 23 E, alt. c. 480 m, 27 June K.-H. Ka, sp. nov. 2008, V. Anton ın 08.39 (BRNM 718679); MycoBank MB 828304 Chuncheon, Dongsan-myeon, Bongmyong-ri, 20 Diagnosis: Gymnopus brunneodiscus differs from August 2007, R. Ryoo KG 139 (BRNM 781444); G. ocior by a light brown to (chestnut) brown pileus 258 R. RYOO ET AL.
broadened at base (up to 8 mm), slightly broadened above, smooth, glabrous except for finely pruinose apex, pale brown with orange to orange-brown tinge (7–8C–E7), apex paler, yellowish white (4A3) when young; with dirty whitish or pale brownish basal tomentum. Context concolorous with surface of basidiomata, hollow in stipe, whitish in pileus, with- out a special smell and with a mild taste. Basidiospores (4.7)5.0–7.0 2.7–4.0 mm, average ¼ 5.6 3.3 mm, E ¼ (1.4)1.6–2.2, Q ¼ (1.5)1.7–1.9, ellipsoid-fusoid, ellipsoid, hyaline, thin-walled. Basidia 20–25 5.5–7.0 mm, 4-spored, clavate. Basidioles 12–28 3.0–7.5 mm, cylindrical, clavate, fusoid. Cheilocystidia 18–68 3.5–6.0(8.0) mm, cylindrical, (narrowly) clavate, irregular, lobed, branched to coralloid, thin-walled. Pleurocystidia absent. Trama hyphae cylindrical or subinflated, thin- to slightly thick-walled, branched, smooth or less frequently minutely incrusted, hyaline, Figure 5. Gymnopus brunneodiscus (BRNM 714970). (A) chei- 3.0–15 mm wide. Pileipellis composed of cells form- locystidia; (B) basidiospores; (C) pileipellis cells. Scale bar ing a Dryophila-structure, cells often relatively slen- ¼ 20 lm. der, irregular, lobate to coralloid, smooth or incrusted, ± thin-walled, 3.0–15 mm wide; incrusta- with a paler, almost whitish margin, a pale brown tion ± hyaline (margin) to (dark) brown (center) in stipe with an orange to orange-brown tinge, and KOH. Pileocystidia absent. Stipitipellis a cutis of larger cheilocystidia, forming an independent clade in cylindrical, parallelly arranged, slightly thick-walled, the nrITS and nrLSU phylogeny. Pileus of G. brunneo- smooth or minutely incrusted, 3.0–5.0(6.0) mm wide discus translucently striate, light brown to (chestnut) hyphae. Caulocystidia absent; scattered adpressed to brown, with almost whitish margin; stipe pale brown suberect, clavate to cylindrical, ± thin-walled ter- with orange tinge to orange-brown with yellowish minal cells present. Clamp connections present in – – m white apex; basidiospores (4.7)5.0 7.0 2.7 4.0 m, all tissues. average ¼ 5.6 3.3 mm; cheilocystidia 18–68 Ecology: On detritus of Larix sibirica, Pinus den- 3.5–6.0(8.0) mm, cylindrical, (narrowly) clavate, siflora, Acer palmatum, Ailanthus glandulosa, irregular, lobed, branched to coralloid; pileipellis of Castanea crenata, Quercus mongolica and Quercus the Dryophila-structure Figures 3 and 5. sp. in mixed forests. Holotype: Republic of Korea. Tean, Mt. Baekhwa, Etymology: brunneus ¼ brown, discus ¼ disk. 0 00 0 00 36 46 16 N, 126 18 54 E, alt. c. 100 m, 10 July 2014, Having a brown-colored pileus center. V. Anton ın 14.73 & K.-H. Ka (BRNM 808975, des- Additional Specimens Examined: Republic of ignated here). Korea. Chuncheon, Dongsan-myeon, Experimental Basidiomata single or in groups. Pileus 15–38 mm forest of Kangwon National University, 37 4604600N, broad, convex-conical to low convex when young, 127 4805900E, alt. c. 210 m, 22 July 2007, V. Anton ın then almost applanate with ± plane center or with 07.102, H.D. Shin & R. Ryoo (BRNM 714970); Ibid., indistinct umbo in small central depression and V. Anton ın 07.108, H.D. Shin & R. Ryoo (BRNM inflexed, later inflexed to straight margin, hygropha- 714974); Cheongju, Sangdang-sanseong Natural nous, (slightly) translucently striate up to half of the Recreational Forest, 36 4004800N, 127 3204000E, alt. c. pileus, smooth, glabrous, light brown to (chestnut) 180 m, 27 August 2015, V. Anton ın 15.112, K.-H. brown (6 C–D5 to 7 D–F6–7or7–8E–F7), with Ka, K.-S. Kim & J.A. Kang (BRNM 808971); paler, almost whitish margin (especially when Hongcheon, Dong-myeon, 37 4100700N, 128 0000600E, young), drying out to pale yellow or light yellow (± alt. 285 m, 17 August 2013, V. Anton ın 13.175 & R. 4A3–4); the pale margin seems to be a constant Ryoo (BRNM 808972); ibid., 37 4200800N, character. Lamellae close, L ¼ 25–45, l ¼ 3–6, emar- 128 0000600E, alt. c. 580 m, 17 August 2013, V. ginate and attached with tooth, ±horizontal, slightly Anton ın 13.173 & R. Ryoo (BRNM 808973); ibid., intervenose, pale yellowish white (3–4A2), some- 37 4101000N, 128 0000200E, alt. c. 310 m, 17 August times with brownish tinge when young, with con- 2013, V. Anton ın 13.180 & R. Ryoo (BRNM colorous, finely pubescent edge. Stipe 808974); Tean, Anmyeon-do, 36 2904700N, 35–70 1–3 mm, cylindrical, sometimes slightly 126 2103600E, alt. 40 m, 11 July 2014, V. Anton ın MYCOBIOLOGY 259
14.83 & K.-H. Ka (BRNM 808976); Chungju, 4. Pileus orangish ochraceous; stipe pale brownish Joryeongsan Natural Forest, 36 4804300N, with paler apex; basidiospores 5.0–6.0(6.5) 128 0204300E, alt. 495 m, 28 August 2015, V. Anton ın 2.7–3.5 mm, average 5.6 3.1 mm, Q ¼ 15.115, K.-H. Ka, S.-K. Kim & J.-A. Kang 1.83 Gymnopus sp. 1 ...... (BRNM 808977). 4 . Pileus ochraceous brownish at center, pale Remarks: Gymnopus brunneodiscus is character- cream at margin; stipe pale cream at apex, ized by having a light to dark brown pileus with a (orange) brown toward base; basidiospores paler margin (which seems to be a constant charac- 5.5–6.5 (2.5)2.7–3.2(3.5) mm, average m ¼ ter), closely placed yellowish white lamellae, a 6.0 3.0 m, Q 2.03...... Gymnopus sp. 2 smooth and glabrous stipe, (4.7)5.0–7.0 2.7–4.0 mm large basidiospores, and cylindrical, (narrowly) cla- 4. Discussion vate, irregular, lobed to coralloid cheilocystidia. Our collection from Tean, Anmyeon-do (BRNM 808976) Species of Gymnopus sect. Levipedes from the differs by having smaller basidiospores Republic of Korea have been distinguished based on ((4.2)4.5–5.5(6.0) 2.5–3.2 mm), and shorter cheilo- their macro- and micromorphology and molecular cystidia (18–35 4.0–7.0 mm). characteristics. The phylogenetic position of Gymnopus dryophilus complex is similar in nrITS 3.2.2. Key to species of Gymnopus sect. Levipedes and nrLSU to that published in previous papers collected in the Republic of Korea [5,18] but the Korean collections form an independ- 1. Pileus ochraceous to (chestnut) brown, with ent clade, a new species, G. dryophiloides. This study constantly almost whitish outer margin; cheilo- shows that G. nubicola Halling is a part of the G. cystidia 18–65 3.5–6.0(8.0) mm, cylindrical, dryophilus group in the nrITS phylogeny, although (narrowly) clavate, irregular, lobed, branched to it has morphologically different characters [5]. The coralloid; pileipellis of the typical Dryophila- separate positions of G. brunneodiscus, Gymnopus structure. Gymnopus brunneodiscus spp. 1, 2 and 3 have been confirmed based on their ...... 1 . Pileus white, light to grayish orange or watery morphological and molecular characterization. ochraceous yellow, ochraceous brownish, light Although Gymnopus spp. 1, 2 and 3 represent both or grayish orange; cheilocystidia up to 38(50) morphologically and phylogenetically separate new 4.5–13(17) mm, more distinctly clavate; pileipel- species, the collected material (only one collection, lis of the typical Dryophila-structure or transi- sometimes only one basidioma) is too poor for for-
ent to a cutis...... 2 mal descriptions of these taxa. 2. Pileus milky white to pale yellow-white with Gymnopus dryophiloides is morphologically and irregularly dispersed brown stains; stipe whitish phylogenetically closely related to G. dryophilus,a to pale yellowish ochraceous; basidiospores species widely distributed throughout Europe [1]. It (6.0)6.5–7.0(7.5) 3.5–4.0(4.5) mm, average differs by a more distinct orange tinge of the stipe, 6.7 3.9 mm Gymnopus sp. 3 sometimes distinctly yellow lamellae, slightly smaller ...... 2 . Pileus light to grayish orange or watery ochra- (shorter) basidiospores and shorter cheilocystidia ceous yellow, ochraceous brownish, light or with a greater variability in shape [1,18]; for a grayish orange; stipe light yellow to light detailed comparison, see Table 2. According to our orange, pale brownish or (orange) brown; basi- phylogenetic studies, it seems that North American diospores (4.0)4.5–6.5 2.5–3.5(4) mm, average collections also form a separate clade. Gymnopus m less than 6.0 3.1 m...... 3 nubicola, described from Ecuador, should differ by a 3. Pileipellis of the typical Dryophila-structure; dark reddish brown pileus when young and fresh, pileus light to grayish orange or watery ochra- eventually becoming brown to light brown, occa- ceous yellow; stipe light yellow to light orange, sionally becoming grayish orange and then mostly sometimes darker in old basidiomata; basidio- at the margin, flesh with a pungent, almost waxy spores (4.0)4.5–6.0(6.5) 2.5–3.5(4.0) mm, smell, and a mild to waxy taste, a brown to dark average ¼ 5.3 3.1 mm; basidia 17–23(25) mm brown stipe toward base, and small, long Gymnopus dryophiloides 10–20 3–5 mm, scattered and inconspicuous, ...... 3 . Pileipellis transient between the Dryophila- irregularly lobed and knobbed cheilocystidia [3]. structure and a cutis; pileus ochraceous brown- However, the phylogenetic placement of this collec- ish, paler toward margin, light or grayish tion supports the proposal by Matta et al. [5]to orange; stipe pale cream and (orange) brown consider it merely a form of an (American) type of toward base or pale brownish; basidiospores “G. dryophilus”. Gymnopus sp. 2 (Yangyang, 5.0–6.5 2.7–3.5 mm, length more than 5.3 mm Micheongol Natural Resort Forest, 26 June 2008, V. – m ı on average; basidia 16 21 m long...... 4 Anton n 08.29, BRNM 718669, Figures 3 and 6) 260 R. RYOO ET AL.
Table 2. Differential macro- and micromorphological characters of G. dryophiloides against similar species. G. dryophiloides G. dryophilus [1,18] Pileus pale to light yellow or orange or watery ochraceous-yellow pale colored, orange-brown or ochraceous brown, then ochraceous-brown, yellow-ochraceous to pinkish ochraceous Lamellae whitish then yellowish white white, cream to yellow Stipe 30–75 1.5–5 mm, concolorous with lamellae at apex, pale 30–120 1–5 mm, yellowish with paler apex, sometimes to light yellow or orange, sometimes darker, light somewhat darker ochraceous brown in basal part orange or grayish orange Basidiospores (4.0)4.5–6.0(6.5) 2.5–3.5(4.0) lm 5.0–7.0(8.0) (2.5)3.0–4.0(4.5) lm Cheilocystidia 17–38(50) 4.5–10(17) lm, variable in shape, clavate, 17–55 4.0–10 lm, (sub)cylindrical, narrowly clavate, (sub)cylindrical, fusoid, mostly lobate, irregular mostly coralloid, also lobate or with apical projections to coralloid
Figure 6. Gymnopus sp. 1 (BRNM 808978). (A) cheilocystidia; (B) basidiospores; (C) pileipellis cells. Scale bar ¼ 20 lm. differs by a centrally ochraceous brownish and mar- ginally pale cream pileus, an (orange) brown stipe Figure 7. Gymnopus sp. 2 (BRNM 718669). (A) cheilocystidia; ¼ l toward base, larger basidiospores, 5.5–6.5 (B) basidiospores; (C) pileipellis cells. Scale bar 20 m. (2.5)2.7–3.2(3.5) mm, Q ¼ 2.03, shorter basidia (18–21 5.5–7.5 mm) and a pileipellis forming a Gymnopus sepiiconicus (Corner) A.W. Wilson, transition between a Dryophila-structure and a cutis. Desjardin & E. Horak, Corner [22] mentioned Gymnopus sp. 3 (Hamyang, Macheon-myeon, 8 July smaller basidiomata (pileus up to 18 mm wide, stipe 2008, V. Anton ın 08.72 & R. Ryoo, BRNM 718701, up to 50 1.5 mm), and slightly smaller basidio- Figures 3 and 7) differs by a milky white to pale yel- spores (4.5–6 2.7–3.5 mm). According to Wilson low-white pileus with irregularly dispersed brown et al. [4], the Indonesian collections of G. sepiiconi- stains, brown stained lamellae, larger basidiospores, cus differ by slightly different basidiospores – – m (6.0)6.5 7.0(7.5) 3.5 4.0(4.5) m, and smaller (4.8–6.4 2.4–4.4 mm) and shorter cheilocystidia – – m cheilocystidia (23 30 6.0 10 m). (20–44 3–7 mm), G. alpinus (Vilgalys & O.K. Mill.) The clade formed by Gymnopus brunneodiscus Anton ın & Noordel. has a dark brown pileus and has 1.00 posterior probabilities and shows one inde- larger, (6.2)6.5–8.5 3.0–4.4 mm basidiospores and pendent group phylogenetically. Gymnopus ocior dif- differently shaped cheilocystidia [1]. Gymnopus fers by a dark red- or orange-brown pileus bicolor A.W. Wilson, Desjardin & E. Horak also has sometimes with yellowish or yellow-red (never whit- a distinctly paler margin, but has larger, ish) margin, a yellow to ochraceous or reddish 5.2–8.0 2.4–3.6 mm basidiospores and differently brown stipe and smaller, 12–45 3.0–9.0 mm cheilo- shaped cheilocystidia [4]. Gymnopus earleae Murrill cystidia [1,18]. In the original description of has a dark brown pileus, but differs by a basally MYCOBIOLOGY 261 m, ] m 2 [ 2.8 m – m 2(4) mm, – 1 2.0 3.5 – G. earleae 30 2.8 – 7 40(50) – orangish buff when young, becoming yellowish orange (ochraceous orange) at the narrowly cylindric to flexuous contorted, rarely diverticulate and rare when present, often collapsed, 24.5 apex and a tawnybelow brown with age fresh, fading to an orangish buff becoming orangish buff – usually absent, inconspicuous 22 Dark brown when young and m, ] 4 m [ m 5.6 m 9.5 – 3.6 – 6.5 3 mm, reddish brown, with white – 2.4 – G. bicolor 2 25.5 8.0 – 42 – broadly clavate, lobulate, smooth beige apex brown, becoming pale orange white margin – 17.5 30 Brown with beige m, m ] m 5.2 l 18 , 11.5
1 Figure 8. Gymnopus sp. 3 (BRNM 718701). (A) cheilocysti- 4.4 [ – – dia; (B) basidiospores; (C) pileipellis cells. Scale bar ¼ 20 lm. 6.0 3.0 6 mm, pale yellow – 2.5 8.5
G. alpinus brown stipe, smaller basidiospores – 30(44)
– – – m 60 (5.6 6.4 2.8 3.5 m) and smaller, usually absent or clavate, simple, irregular to coralloid at apex, orange-yellow to orange-brown below brown, uniform, or only slightly paler at very margin – 16.5 30 Very dark red- to purplish inconspicuous (and then 24.5–30 2.0–2.8 mm) cheilocystidia [6]; for a comparison, see Table 3. Gymnopus sp. 1 (Hocheong, 4 Jul 2014, V. Anton ın ]
4 14.45, R. Ryoo & K.-H. Ka, BRNM 808978, Figures [
m (6.2)6.5 3 and 8) differs by an almost entirely orangish l m, irregularly 4.4 ochraceous pileus with a paler outer margin, yellow l – 7 2 mm, brownish against similar species. – – 2.4 lamellae, a pale brownish stipe, smaller basidio- 3 1 G. sepiiconicus spores, 5.0–6.0(6.5) 2.7–3.5 mm, and smaller, 6.4 44 52 – cylindrical to obtuse clavate, contorted, lobulate orange to brown orbuff, creamy with orange beige apex beige yellow to white margin in age – – 14–30 6.0–11(13) mm, clavate, subcylindrical chei- 20 4.8 40 Brown to dark brown with locystidia, which are only irregular, lobate or with projection(s). G. brunneodiscus ] m l 18 , Acknowledgement 1 m, clavate, [ l ı 12 The authors thank J.W. Jongepier (Vesel nad Moravou, 3.5(4.0) – 5 mm, yellow to –
– Czech Republic) for linguistic corrections of 6.0 2 6.5(7.0) – the manuscript. 60 60 subcylindrical or subutriform, often lobed, branched, coralloid or with apical projections (2.5)2.75 ochraceous or reddish brown pallescent to reddish yellow or pinkish brownish – – 16 20 Whitish or yellowish lamellae White White or cream colored White Pale orangish yellow, Dark red- or orange-brown, Disclosure statement No potential conflict of interest was reported by m (5.0)5.5
m, the author(s). l l 4.0 – 2.7
6.0(8.0) Funding – 3 mm, pale brown – 3.5 1 7.0 G. brunneodiscus G. ocior – This research was supported by a Korea Research 68 70 cylindrical, (narrowly) clavate, irregular, lobed, branched to coralloid, with orange tinge to orange-brown, with paler, yellowish white apex when young sometimes with brownish tinge when young brown, with paler, almost whitish margin Foundation grant funded by project FP0801-2010-01 of – – the National Institute of Forest Sciences (NIFoS). The laboratory studies of the second author were supported
Differential macro- and micromorphological characters of by the Moravian Museum funded by the Ministry of Culture of the Czech Republic as part of its long-term conceptual development program for research institutions Cheilocystidia 18 Basidiospores (4.7)5.0 LamellaeStipe Pale yellowish white, 35 Table 3. Pileus Light brown to (chestnut) [DKRVO, ref. MK000094862]. 262 R. RYOO ET AL.
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