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Research Article
Two new species of Gymnopus sect. Levipedes (Omphalotaceae, Agaricales) from Central and North China
expand article infoJia-Jun Hu‡§, Yong-Lan Tuo§, Zheng-Xiang Qi§, Dong-Hua Jiang‡, Yu Li§, Bo Zhang§
‡ Zhejiang Normal University, Jinhua City, China
§ Jilin Agricultural University, Changchun City, China

Corresponding author: Dong-Hua Jiang ( jdh@zjnu.cn )

Corresponding author: Yu Li ( fungi966@126.com )

Corresponding author: Bo Zhang ( zhangbofungi@126.com )

Academic editor: Thorsten Lumbsch
© 2025 Jia-Jun Hu, Yong-Lan Tuo, Zheng-Xiang Qi, Dong-Hua Jiang, Yu Li, Bo Zhang.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Hu J-J, Tuo Y-L, Qi Z-X, Jiang D-H, Li Y, Zhang B (2025) Two new species of Gymnopus sect. Levipedes (Omphalotaceae, Agaricales) from Central and North China. MycoKeys 118: 19-34. https://doi.org/10.3897/mycokeys.118.143277
Open Access

Abstract

The genus Gymnopus has a long research history and is known for its high species diversity worldwide. However, its species diversity in China remains poorly understood. Through a combination of detailed morphological studies and phylogenetic analysis, this study described two new species from Northeast and Central China, Gymnopus biyangensis and Gymnopus sinodryophilus, both belonging to Gymnopus sect. Levipedes. Gymnopus biyangensis is characterized by basidiomata appearing in summer and originating from broad-leaved forests, a dark reddish-brown pileus, a cylindrical to clavate stipe, and clavate to cylindrical, diverticulate cheilocystidia. Gymnopus sinodryophilus differs by the yellowish-white to light brown basidiomata arising from coniferous and broad-leaved mixed forests, coralloid but inflated pileipellis terminal cells, and an apex of cheilocystidia not coralloid. Additionally, a key to the reported species of Gymnopus sect. Levipedes in China is provided.

Key words:

Cheilocystidia, ecology, Gymnopus dryophilus complex

Introduction

Gymnopus (Pers.) Gray has a long research history dating back to 1801 (Persoon 1801) and plays a significant role in various ecosystems (Mata et al. 2006). In many countries, including China and elsewhere, certain species of Gymnopus are also treated as food resources (Hu and Zhang 2023). As with many macrofungi, Gymnopus (previously classified under the genus Collybia) was first studied in Europe, with significant contributions made by European mycologists (Persoon 1801; Fries 1821; Antonín et al. 1997; Antonín and Noordeloos 1997). Over time, this research enthusiasm spread to other regions (Cooper and Leonard 2013; Antonín et al. 2014; Coimbra et al. 2015; Deng 2016; Oliveira et al. 2019; Hu et al. 2022a, b), particularly North America, where many studies have been conducted (Halling 1996; Desjardin et al. 1999; Mata et al. 2006; Petersen and Hughes 2014).

However, compared to other genera, such as Agaricus L. and Cantharellus Adans. ex Fr., Gymnopus has not been as thoroughly studied. This discrepancy in research interest is partly due to regional differences in mycologists’ areas of focus, which, have led to significant research gaps between Gymnopus and more widely studied genera. For example, approximately 300 species of Gymnopus have been described globally (Hu et al. 2022a, b), with the majority of species documented in North America, Europe, and Asia (Sun et al. 2021b; Hu and Zhang 2023). In contrast, fewer species have been reported in Oceania and Africa (Hu and Zhang 2023).

In recent decades, Asia has emerged as a new hotspot for Gymnopus research, evidenced by the discovery of new species or new combinations (Hu and Zhang 2023). This indicates an increasing recognition of the genus’ biodiversity and ecological importance. Nevertheless, the full extent of Gymnopus species diversity remains underappreciated and warrants further exploration.

Subsection Levipedes is characterized by a stipe that is smooth, polished, or pubescent; a pileipellis that typically forms an entangled (never radially oriented) trichoderm, consisting of inflated, often lobed elements or coralloid “dryophila-type” structures; trama and its elements are non-dextrinoid, and usually saprophytic, commonly found in coarse humus, forest litter, or on decaying wood (Halling 1996; Antonín and Noordeloos 2010; Oliveira et al. 2019). In this subsection, some species belonging to the Gymnopus dryophilus (Bull.) Murrill complex have historically been treated as food resources in Northeast and Southwest China (Hu and Zhang 2023). Despite this, recent studies have shown that the species complex, Gymnopus dryophiloides, can cause diarrhea in humans (Ma et al. 2024). Furthermore, due to the similarity in appearance, difficulties remain in identifying the species of this complex. Although Vilgalys and Miller (1983, 1987a, b) and Antonín et al. (2013) have carefully studied the North American or European G. dryophilus species complex and identified key characteristic features, all these results were based on the North American or European specimens. Additionally, there is still a lack of comprehensive study on the complex outside of North America and Europe.

Looking back at the research history, it is evident that taxonomic studies are mainly focused on Europe and North America, leaving a significant gap on other continents. As research on Gymnopus deepens, taxonomic issues and questions about the classification of the genus emerge, highlighting the need for further attention to this group. Consequently, taxonomic research on Gymnopus has begun in China. Specimens of the G. dryophilus complex collected from Henan and Jilin Provinces are studied in detail here. As a result, two new species belonging to Gymnopus sect. Levipedes are described and illustrated. This study enriches our understanding of the species diversity of Gymnopus and provides valuable insights for future studies.

Materials and methods

Specimen collection

The specimens used in this study were collected between 2020 and 2023 in Henan and Jilin Provinces, China. All specimens were photographed in situ, with an emphasis on capturing basidiomata at various stages of development. Subsequently, three or more basidiomata were collected for detailed morphological and molecular analysis. The morphological characteristics, including size, color, and odor, were documented. The color references followed Flora of British Fungi: Color Identification Chart (Royal Botanic Garden 1969). A clean tissue sample from each specimen was dried using allochroic silica gel for DNA extraction. The specimens were dried in an electric oven at approximately 45 °C.

Identification

The recognition and description of macro-characteristics were based on field notes and photographs. Dried specimens were rehydrated in 94% ethanol and subsequently mounted in 3% potassium hydroxide (KOH), 1% Congo Red, or Melzer’s Reagent for examination. Structures such as basidiospores, basidia, and cheilocystidia were observed using a Zeiss Axio Lab. A1 microscope. For each specimen, a minimum of 40 measurements were taken from at least two different basidiomata. The size of basidiospores is expressed as length × width (L × W). To account for size variation, 5% of the measurements from each end of the range were excluded, and the final measurements are given as (a) b × c (d). Q represents the ratio of L to W for each studied specimen, while Qm denotes the average Q value ± standard deviation. The examined specimens are deposited in the Herbarium of Mycology at Zhejiang Normal University (ZNU-F).

DNA extraction, PCR, and sequencing

Total DNA was extracted from dried materials using the NuClean Plant Genomic DNA Kit (Kangwei Century Biotechnology Company Limited, Beijing, China), following the manufacturer’s instructions. The internal transcribed spacer (ITS) region, nuclear large ribosomal subunits (nLSU), and translation elongation factor (tef-1α) loci were selected for phylogenetic analysis. The primer pairs ITS4-ITS5 (Gardes and Bruns 1993), LROR-LR5/LR7 (Vilgalys and Hester 1990; Cubeta et al. 1991), and 983F-1567R (Rehner and Buckley 2005) were used to amplify the ITS, nLSU, and tef-1α, respectively.

PCR reactions (25 μL) were prepared as follows: 8.5 μL of dd H2O, 12.5 μL of 2 × Taq PCR MasterMix, 1 μL of each primer, and 2 μL of DNA sample. The reaction conditions were based on those described by Coimbra et al. (2015) for ITS, Ryoo et al. (2020) for nLSU, and Xu et al. (2021) for tef1-α. PCR products were visualized under UV light following electrophoresis on 1% agarose gels stained with ethidium bromide. The PCR products were then sent to Hangzhou Huada-Qinglan Innovation Technology Co., Ltd. for sequencing, using the Sanger method. The new sequences were deposited in GenBank (http://www.ncbi.nlm.nih.gov/genbank), and the detailed sequence information is provided in Table 1.

Table 1.
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Voucher/specimen numbers, country, and GenBank accession numbers of the specimens included in this study. Sequences produced in this study are in bold and obtained from type materials marked as T.

Scientific name Country Specimen/Voucher numbers GenBank Accession Numbers
ITS nLSU tef1-α
Collybiopsis juniperinus USA TENN59540 AY256708 KY019637 –
Collybiopsis obscuroides Norway GB-0150514 KX958399 KX958399 –
Collybiopsis subnuda USA TENN-F-61138 KY026667 FJ750262 –
Gymnopus abruptibulbus nom. prov. China HMJAU61050 OQ597084 – –
Gymnopus alkalivirens USA TENN51249 DQ450000 – –
Gymnopus alliifoetidissimus China GDGM76695 MT023344 MT017526 –
Gymnopus alpicola Spain BRNM705055 MK278102 MK278102 –
Gymnopus alpinus Latvia CB16251 JX536168 – JX536191
Gymnopus androsaceus Russia TENN-F-59594 KY026663 KY026663 –
Gymnopus androsaceus France CBS239.53 MH857174 MH868713 –
Gymnopus aquosus Czech Republic BRNM665362 JX536172 – JX536192
Gymnopus atlanticus (T) Brazi URM87728 KT222654 KY302698 –
Gymnopus aurantiipes – AWW118 AY263432 AY639410 –
Gymnopus austrosemihirtipes (T) Indonesia SFSU-AWW65 AY263422 – –
Gymnopus barbipes USA TENN67855 KJ416269 NG_059733 –
Gymnopus bicolor – AWW116 AY263423 AY639411 –
Gymnopus bisporus Spain BCN-SCM B-4065 JN247551 JN247555 –
Gymnopus biyangensis (T) China ZNU-F-001 PQ651934 PQ651940 PQ661922
Gymnopus biyangensis China ZNU-F-002 PQ651935 PQ651941 PQ661923
Gymnopus biyangensis China ZNU-F-003 PQ651936 PQ651942 PQ661924
Gymnopus brassicolens Russia TENN55550 DQ449989 – –
Gymnopus brunneiniger (T) Mexico XAL-Cesar 49 MT232389 NG-075396 –
Gymnopus brunneodiscus Korea BRNM 808975 MH589975 MH589991 –
Gymnopus campanifomipileatus nom. prov. China HMJAU61027 OQ597064 OQ594474 –
Gymnopus catalonicus Spain BCN-SCM B-4057 JN247552 JN247556 –
Gymnopus ceraceicola (T) New Zealand PDD87181 KC248405 – –
Gymnopus changbaiensis (T) China HMJAU60300 OM030272 OM033387 –
Gymnopus cremeostipitatus (T) Korea BRNM747547 KF251071 KF251091 –
Gymnopus cystidiosus (T) China HMJAU60992 ON259024 ON259036 –
Gymnopus densilamellatus (T) Korea BRNM714927 KP336685 KP336694 –
Gymnopus dryophilus Czech Republic BRNM695586 JX536143 – JX536196
Gymnopus dryophilus Japan Duke31 DQ480099 – –
Gymnopus dryophioides (T) South Korea BRNM781447 MH589967 MH589985 –
Gymnopus dysodes USA TENN-F-61125 KY026666 KY026666 –
Gymnopus earleae USA TENN-F-59140 DQ449994 KY019634 –
Gymnopus efibulatus (T) China HGASMF01-7052 OM970865 OM970865 –
Gymnopus epiphyllus (T) China HMJAU60990 ON259030 ON259038 –
Gymnopus erythropus Czech Republic BRNM714784 JX536136 – JX536183
Gymnopus fagiphilus Czech Republic BRNM707079 JX536129 – JX536209
Gymnopus foetidus USA TENN-F-65806 KY026682 KY026682 –
Gymnopus fuscopurpureus Spain BRNM-809119 MZ542559 MZ542563 –
Gymnopus fusipes Austria TENN59300 AF505777 – –
Gymnopus fusipes France TENN59217 AY256710 AY256710 –
Gymnopus globulosus (T) China HMJAU60307 OM030269 OM033406 –
Gymnopus graveolens France FF17084 MH422573 MH422572 –
Gymnopus hakaroa (T) New Zealand PDD87315 KC248410 – –
Gymnopus hemisphaericus nom. prov. China HMJAU61077 OQ597057 OQ594467 –
Gymnopus hybridus Italy BRNM695773 JX536177 – JX536208
Gymnopus imbricatus (T) New Zealand PDD95489 KC248390 – –
Gymnopus impudicus Russia TENN60094 KJ416263 – –
Gymnopus indoctoides Singapore AY263424 AY639419 – –
Gymnopus inexpectatus Italy – EU622905 EU622906 –
Gymnopus inusitatus Spain BCN-SCM B-4058 JN247553 JN247557 –
Gymnopus inusitatus var. cystidiatus (T) Hungary BRNM737257 JN247550 JN247554 JX536179
Gymnopus iocephalus USA TENN52970 DQ449984 KY019630 –
Gymnopus iodes (T) China HGASMF01-10068 OM970869 OM970869 –
Gymnopus irresolutus Sao Tome SFSU-DED-8209 MF100973 – –
Gymnopus junquilleus (T) USA TENN55224 NR_119582 – –
Gymnopus kauffmanii USA DUKE230 DQ450001 – –
Gymnopus lachnophyllus USA NAMA2015-320 MH910564 – –
Gymnopus lanipes (T) Spain BRNM670686 JX536137 – JX536205
Gymnopus loiseleurietorum Sweden URM 90060 KY321571 KY321572 –
Gymnopus longisterigmaticus (T) China HMJAU60288 OM030282 OM033403 –
Gymnopus longistipes (T) China HMJAU61076 PP646156 PP646168 PP654450
Gymnopus longus (T) China HMJAU60291 OM030285 OM033400 –
Gymnopus macropus Costa Rica TENN58619 DQ449979 – –
Gymnopus macrosporus (T) China HMJAU60294 OM030266 OM033397 –
Gymnopus montagnei Brazil URM87715 KT222652 – –
Gymnopus neobrevipes (T) USA TENN-F-14505H1 MH673477 MH673477 –
Gymnopus nubicola Costa Rica NYBG REH 8290 AF505781 – –
Gymnopus ocior Czech Republic BRNM699795 JX536166 – JX536188
Gymnopus omphalinoides (T) China GDGM 78318 MW134044 MW134730 –
Gymnopus pallipes (T) China GDGM81513 MW582856 – –
Gymnopus polyphyllus USA TENN59455 AY256695 – –
Gymnopus pubipes Spain AH26931 MZ542558 MZ542562 –
Gymnopus pygmaeus Brazil URM90003 KX869966 KY088273 –
Gymnopus salakensis Indonesia SFSU-AWW29 AY263447 – –
Gymnopus schizophyllus (T) China GDGM 77165 MW134043 MW134729 –
Gymnopus semihirtipes USA TENN-F-07595 OK376741 – –
Gymnopus sepiiconicus Indonesia SFSU-AWW126 AY263449 – –
Gymnopus similis (T) Korea BRNM766739 KP336692 KP336699 –
Gymnopus sinodryophilus China ZNU-F-004 PQ651937 PQ651943 PQ661925
Gymnopus sinodryophilus (T) China ZNU-F-005 PQ651938 PQ651944 PQ661926
Gymnopus sinodryophilus China ZNU-F-006 PQ651939 PQ651945 PQ661927
Gymnopus spadiceus (T) China HMJAU61205 PP646160 PP646172 PP654454
Gymnopus spongiosus USA TENN-F-68184 KY026706 KY026706 –
Gymnopus striatipileatus (T) China HMJAU61073 PP646166 PP646178 –
Gymnopus striatus (T) China HMJAU60297 OM030263 OM033384 –
Gymnopus strigosipes (T) China HMAS295796 OM970874 OM970874 –
Gymnopus subdensilamellatus (T) China HMJAU60997 ON259032 ON259042 –
Gymnopus subpolyphyllus (T) China HMJAU60999 ON259028 ON259043 –
Gymnopus subsulphureus USA TENN56321 DQ449972 – –
Gymnopus subsupinus New Zealand PDD96595 KM975399 KM975375 –
Gymnopus talisiae (T) Brazil URM87730 KT222655 KX958401 –
Gymnopus tiliicola (T) China HMJAU60305 OM030275 OM033393 –
Gymnopus tomentosus (T) China HMJAU60303 OM030278 OM033390 –
Gymnopus trabzonensis (T) Turkey KATO Fungi 3375 KT271754 – –
Gymnopus variicolor (T) Korea BRNM714959 LT594121 KP348011 –
Gymnopus viridiscus (T) China HMJAU61202 PP646159 PP646171 PP654453
Gymnopus vitellinipes (T) Indonesia SFSU-AWW127 AY263429 AY639432 –
Marasmius aurantioferrugineus South Korea BRNM714752 FJ904962 MK278334 –
Marasmius brunneospermus (T) South Korea KPM-NC0005011 FJ904969 FJ904951 –

Phylogenetic analysis

Based on the BLASTn results and morphological similarities, sequences related to these samples were collected (Table 1). A combined dataset of ITS, nLSU, and tef-1α fragments, consisting of 85 sequences obtained from the type species of Gymnopus and Marasmius Fr., was used for phylogenetic analysis. Species belonging to Marasmius, Marasmius aurantioferrugineus Hongo and Marasmius brunneospermus Har. Takah., were selected as outgroups (Hu et al. 2024).

Each gene region in the dataset was aligned using MAFFT 7.490 (Katoh and Standley 2013) and subsequently manually inspected in BioEdit 7.0.5.3 (Hall 1999). The alignments of the ITS, nLSU, and tef-1α sequences were then combined through PhyloSuite 1.2.2 (Zhang et al. 2020). A partition homogeneity test (PHT) (Farris et al. 1994) was performed on the multi-gene dataset with PAUP 4.0b10 (Swofford 2002), employing 1000 homogeneity replicates. The best-fit evolutionary model was estimated using ModelFinder (Kalyaanamoorthy et al. 2017). Bayesian inference (BI) was applied for phylogenetic analysis, utilizing MrBayes 3.2.6 with a general time-reversible DNA substitution model and gamma distribution rate variation across the sites (Ronquist and Huelsenbeck 2003). Four Markov chains were run for two independent runs, starting from random trees, until the split frequency value fell below 0.01. Trees were sampled every 100 generations, with the first 25% discarded as burn-in. The remaining trees were used to construct a 50% majority consensus tree and calculate the Bayesian posterior probabilities (PP). Maximum likelihood (ML) analysis was performed using RaxmlGUI 2.0.10 (Edler et al. 2021) with 1000 bootstrap (BS) replicates to search for the optimal topology. The resulting trees were visualized using FigTree 1.4.4 (http://tree.bio.ed.ac.uk/software/figtree/, accessed 25 October 2020).

Results

Phylogenetic analysis

A total of 18 new sequences (six per locus) were obtained from six samples in this study. In the combined dataset, 189 sequences derived from three gene loci (ITS, nLSU, and tef-1α) from 102 samples were used for phylogenetic analysis. The best-fitting model for BI was GTR+F+I+G4, while the GTRGAMMA model was applied for ML (Vizzini et al. 2015). The Bayesian analysis was run for four million generations, resulting in an average standard deviation of split frequencies of 0.004528. The same dataset and alignment were also analyzed using the ML method. Both phylogenetic analyses yielded a similar topology, which is depicted in Fig. 1.

Figure 1. 

Maximum likelihood analysis generated from the combined ITS, nLSU, and tef-1α dataset of genus Gymnopus. Bootstrap values (BS) ≥ 75% from ML analysis and Bayesian posterior probabilities (PP) ≥ 0.80 are shown on the branches. Newly sequenced collections are indicated in bold, and the type specimens are denoted by (T).

Our phylogenetic analysis revealed that species of the G. dryophilus complex form a distinct clade, which is sister to the Gymnopus erythropus (Pers.) Antonín, Halling & Noordel. complex. Two newly proposed species are independently positioned within the genus Gymnopus, with strong phylogenetic evidence.

Taxonomy

Gymnopus biyangensis J.J. Hu, B. Zhang, X. Li & Y. Li, sp. nov.

Fungal Names: FN 572234
Figs 2A, 3

Etymology.

Refers to the location of type material.

Diagnosis.

[English] This species is characterized by the basidiomata that appear in summer and originate from broad-leaved forests, dark reddish-brown pileus, cylindrical to clavate stipe, clavate to cylindrical cheilocystidia with a narrowly protruding apex.

Figure 2. 

The habit of Gymnopus spp. described in this study: A Gymnopus biyangensis B Gymnopus changchunensis. Scale bars: 1 cm.

Type.

China. Henan Province • Zhumadian City, Biyang County, Mingzhuang Village, 10 July 2021, Jia-Jun Hu, Bo Zhang, and Xiao Li, ZNU-F-001 (Collection No.: Hu 769), holotype.

Figure 3. 

Morphological characteristics of Gymnopus biyangensis (ZNU-F-001) A basidiomata B pileipellis elements C basidiospores D cheilocystidia E basidia. Scale bars: 1 cm (A); 5 µm (B–E).

Description.

Basidiomata medium-sized. Pileus 3.3–4.4 cm in diameter, applanate-convex, reddish brown to brown, smooth, glabrous; margin entire, wavy to upturned, dark reddish-brown to light brown. Context thin, yellow to light brown, freshy, odorless. Stipe 2.0–5.0 cm long and 0.5–1.3 cm wide, central, cylindrical to clavate, smooth, light yellow, occasionally with reddish brown tones or slight spots at the base. Lamellae adnate to adnexed, close, yellowish brown to light brown, unequal. Occurrence in leaf litter.

Basidiospores 5.0–6.0 × 3.0–4.0 µm, Q = (1.25)1.30–1.93, Qm = 1.64 ± 0.19, elliptic, hyaline, smooth, inamyloid, thin-walled. Basidia (12)16–23 × 3–6 µm, clavate to cylindrical, 2- or 4-spored, hyaline, smooth, thin-walled. Cheilocystidia (16)17–28(29) × 3–6 µm, cylindrical to clavate with mamiform, often longer apical projections, less commonly weakly coralloid, hyaline, smooth, thin-walled. Pleurocystidia and caulocystidia not observed. Pileipellis a “dryophila-type” cutis, 8–14 µm wide, hyaline, smooth, thin-walled. Clamp connections present in all tissues.

Habit, habitat, and distribution.

Scattered to gregarious. Saprotrophic, with humicolous habitat, found in broad-leaved forests. So far, it is only known from Henan Province, China.

Other specimens examined.

China. Henan Province • Zhumadian City, Biyang County, Mingzhuang Village, 10 July 2021, Jia-Jun Hu, Bo Zhang, and Xiao Li, ZNU-F-002 (Collection No.: Hu 770) • Zhumadian City, Biyang County, Mingzhuang Village, 10 July 2022, Jia-Jun Hu, Bo Zhang, and Xiao Li, ZNU-F-003 (Collection No.: Hu 775).

Note.

This species is characterized by the basidiomata occurring in the summer, a dark reddish-brown pileus, a cylindrical to clavate stipe, an apex of cheilocystidia that is not diverticulate or lobate.

Gymnopus biyangensis is similar to species in the G. erythropus complex due to the dark reddish-brown pileus. However, this species differs from G. erythropus by the light yellow and cylindrical to clavate stipe, smaller basidiospores, and a non-encrusted pileipellis. Gymnopus biyangensis can be distinguished from G. fagiphilus by the smooth and light-yellow stipe and smaller basidiospores.

Gymnopus sinodryophilus J.J. Hu, B. Zhang & Y. Li, sp. nov.

Fungal Names: FN 572235
Figs 2B, 4

Etymology.

Refers to the species similar to G. dryophilus.

Diagnosis.

[English] This species is characterized by the yellowish-white to light-brown basidiomata, arisen from coniferous and broad-leaved mixed forest, coralloid but inflated pileipellis elements, and a non-coralloid apex of cheilocystidia.

Figure 4. 

Morphological characteristics of Gymnopus sinodryophilus (ZNU-F-003) A basidiomata B pileipellis elements C basidiospores D basidia E cheilocystidia. Scale bars: 1 cm (A); 5 µm (B–E).

Type.

China. Jilin Province • Changchun City, Jingyue District, Mt. Lianhua, 09 August 2021, Jia-Jun Hu and Bo Zhang, ZNU-F-005 (Collection No.: Hu 809), holotype.

Description.

Basidiomata small to medium. Pileus 1.0–4.4 cm in diameter, applanate-hemispheric to convex, light yellow to light brown, darker at center, brown, occasionally with brown spots, with an umbo sometimes, smooth, glabrous; margin entire, involute, yellowish white to light yellow. Context thin, fresh, white to light yellow, odorless. Stipe 1.3–8.8 cm long and 0.3–0.9 cm wide, central, clavate, light brown to brown, paler downwards, becoming light yellow to yellowish white, striped, smooth, glabrous, fistulose, fibrous. Lamellae adnexed, close to crowded, yellow to light brown, unequal. Occurrence in leaf litter in mixed forest.

Basidiospores (4.0)5.0–6.0(6.2) × 3.0–3.8 µm, Q = (1.33)1.39–2.00(2.07), Qm = 1.67 ± 0.18, elliptic, hyaline, smooth, inamyloid, thin-walled. Basidia (12)13–23 × 4–8 µm, clavate to cylindrical, 2- or 4-spored, hyaline, smooth, thin-walled. Cheilocystidia (11)13–27(31) × 3–8 µm, cylindrical to clavate, umbonate-mamiform with a short apical projection, occasionally forked, hyaline, smooth, thin-walled. Pleurocystidia and caulocystidia not observed. Pileipellis a “dryophila-type” cutis, (5)7–10(12) µm wide, hyaline, smooth, thin-walled. Clamp connections present in all tissues.

Habit, habitat, and distribution.

Scattered to gregarious. Saprotrophic, with humicolous habitat, found in mixed forests. So far, only known from Jilin Province, China.

Other specimens examined.

China. Jilin Province • Yanbian Korean Autonomous Prefecture, Antu County, Erdaobaihe Town, 26 June 2021, Jia-Jun Hu and Bo Zhang, ZNU-F-006 (Collection No.: Hu 743) • Changchun City, Jingyue District, Jingyuetan National Forest Park, 08 August 2021, Jia-Jun Hu, Bo Zhang, ZNU-F-004 (Collection No.: Hu 807) • Changchun City, Jingyue District, Jingyuetan National Forest Park, 08 August 2021, Jia-Jun Hu, Bo Zhang, ZNU-F-007 (Collection No.: Hu 811) • Changchun City, Jingyue District, Jingyuetan National Forest Park, 08 August 2021, Zheng-Hao Zhang, Jia-Jun Hu, Bo Zhang, ZNU-F-008 (Collection No.: Hu 892).

Note.

This species is characterized by the yellowish-white to light-brown basidiomata, which arise in summer; coralloid but inflated pileipellis elements; and a non-diverticulate apex of cheilocystidia.

This species is extremely similar to G. dryophilus due to the analogous morphology. However, this species differs from G. dryophilus by appearance in summer, smaller basidiospores, and the apex of cheilocystidia not being diverticulate.

Key to the reported species of Gymnopus subsect. Levipedes in China

1 Basidiomata with red stipe 2
– Basidiomata with yellow stipe 10
2 Stipe covered with dense hairs at the base 3
– Stipe smooth, or covered with sparse hairs at the base G. erythropus
3 Basidia sterigmata extremely long 4
– Basidia sterigmata short 6
4 Stipe smooth in upper part 5
– Stipe covered with brown pruina on the upper part G. longus
5 Pileus pale color, stipe color uneven G. longisterigmaticus
– Pileus dark color, stipe color uniform G. macrosporus
6 Growing on the deciduous layer or rotten branches 7
– Grows at the base of Tilia sp. G. tiliicola
7 Pileus pale color, near white G. tomentosus
– Pileus deep color 8
8 Stipe covered with longitudinal striate G. striatus
– Stipe without longitudinal striate 9
9 Pileipellis a cuits, typically “dryophila type” G. changbaiensis
– Pileipellis layered, hyphae inflated to spherical to prolate G. globulosus
10 Apex of cheilocystidia diverticulate 11
– Apex of cheilocystidia not diverticulate 19
11 Basidiomata marasmioid G. striatipileatus
– Basidiomata collybioid or tricholomatoid 12
12 Cheilocystidia absent G. longistipes
– Cheilocystidia present 13
13 Caulocystidia present G. inexpectatus
– Caulocystidia absent 14
14 Pileus green G. viridiscus
– Pileus not green 15
15 Stipe cylindrical or clavate 16
– Stipe enlarged at base 18
16 Stipe light red G. aquosus
– Stipe light yellow to yellow 17
17 Stipe covered with tomentose G. brunneodiscus
– Stipe smooth G. dryophilus
18 Stipe pale yellow, basidiospores smaller than 6 µm G. dryophiloides
– Stipe light yellow to reddish brown, basidiospores larger than 6 µm G. ocior
19 Basidiomata and lamellae light yellow G. sinodryophilus
– Basidiomata reddish brown, lamellae light reddish brown G. biyangensis

Discussion

In this study, two new species within the G. dryophilus complex are proposed. Gymnopus biyangensis is characterized by summer-fruiting basidiomata found in broad-leaved forests, a dark reddish-brown pileus, a cylindrical to clavate stipe, and clavate to cylindrical cheilocystidia with a long apical projection. In contrast, Gymnopus sinodryophilus is characterized by yellowish-white to light-brown basidiomata that arise from coniferous and broad-leaved mixed forests, coralloid but inflated pileipellis elements, and cheilocystidia with a short apical projection.

The sect. Levipedes is divided into two subsections: subsect. Alkalivirentes Antonín & Noordel. and subsect. Levipedes Antonín & Noordel., based on whether the mycelium turns green in potassium hydroxide (KOH) and ammonium hydroxide (NH4OH) (Halling 1981; Antonín and Noordeloos 1997).

Species from the G. erythropus, G. fagiphilus (Velen.) Antonín, Halling & Noordel., and G. dryophilus complexes primarily comprise subsect./ Levipedes. Our previous work demonstrated that the G. dryophilus complex species clearly differs from the G. erythropus complex species, particularly in terms of seasonal occurrence and the shape of cheilocystidia (Hu et al. 2022b, Hu 2023). The species of subsect. Levipedes are characterized by smooth, polished, or pubescent stipes; pileipellis is typically an entangled trichoderm (never radially oriented), composed of inflated, often lobed elements or coralloid, “dryophila-type” structures; trama and its elements are non-dextrinoid (Halling 1996; Antonín and Noordeloos 2010; Oliveira et al. 2019). However, in our study, G. sinodryophilus exhibited branched but not broadened pileipellis elements, which contrasts with the established boundaries. Similar deviations have been observed in Gymnopus globulosus J.J. Hu, Y.L. Tuo, B. Zhang & Yu Li, Gymnopus earleae Murrill, and Gymnopus kauffmanii (Halling) Halling, etc. These findings suggest that the current limits and taxonomic framework of subsect. Levipedes require further examination.

Although the taxonomic history of Gymnopus (formerly Collybia) spans over two centuries, research on this genus remains considerably behind other groups, such as Amanitaceae (Cui et al. 2018), boletes (Wu et al. 2016), and Ganodermataceae (Sun et al. 2022). Although some species are edible or possess potential applications, such as in wastewater treatment (Sun et al. 2021a) and biological control (Wu 2016), they have not gained the same recognition as culinary fungi, for example, Morchella Dill. ex Pers. or Tuber P. Micheli ex F.H. Wigg. This lack of prominence may be a key reason why this genus has been overlooked in research. To truly understand this group, greater attention must be devoted to its study.

Acknowledgments

The authors would like to thank Mr. Zheng-Hao Zhang for specimen collection. The authors would like to express their gratitude to the anonymous reviewer(s).

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This study is funded by the National Natural Science Foundation of China (32400011), the Youth Doctoral Program of Zhejiang Normal University (2023QB043), and the Zhejiang Normal University Doctoral Initiation Fund (31970020).

Author contributions

Conceptualization: DHJ, YL, and BZ; Data curation: JJH; Formal analysis: JJH and BZ; Funding acquisition: JJH; Investigation: JJH, ZXQ and YLT; Methodology: JJH, BZ and DHJ; Project administration: BZ; Resources: JJH and BZ; Software: JJH; Supervision: DHJ, YL and BZ; Validation: JJH and BZ; Visualization: JJH; Writing – original draft: JJH; Review, and editing: JJH and BZ.

Author ORCIDs

JiaJun Hu https://orcid.org/0000-0002-7562-7612

Yong-Lan Tuo https://orcid.org/0000-0001-6019-1038

Zheng-Xiang Qi https://orcid.org/0000-0002-0037-9407

Yu Li https://orcid.org/0000-0003-4719-7210

Bo Zhang https://orcid.org/0000-0001-9508-8188

Data availability

All of the data that support the findings of this study are available in the main text.

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