Aspergillaceae was established by Link (1826), with Aspergillus P. Micheli ex Haller designated as the type genus. Houbraken and Samson (2011) reinstated this family based on multi-locus phylogeny and recognized seven distinct clades. Some species in this family can tolerate extreme conditions, including high sugar and salt concentrations, low or high temperatures, low acidity, and low oxygen levels (Houbraken et al. 2014; Zhang et al. 2021). Aspergillaceae species are predominantly saprobic and commonly occur in soil, although some are plant and animal pathogens (Houbraken et al. 2014). Many species in this family are of great economic importance and are widely used in food fermentation, biotechnology, and biomedicine (Mogensen et al. 2009; Perrone et al. 2011; Houbraken et al. 2014). Morphologically, Aspergillaceae members are characterized by asci within cleistothecia or stromata, or enclosed by Hülle cells, and typically have oblate to ellipsoidal ascospores with a distinct furrow or slit (Houbraken and Samson 2011; Houbraken et al. 2014). Conidia are mostly formed on lanceolate, flask-shaped, or cylindrical phialides (Houbraken et al. 2014). Aspergillaceae is the most abundant family reported from cave environments, with 13 genera comprising over 315 species worldwide (Suppl. material 1: table S2).

The genus Aspergillus was established by Haller (1768), with A. glaucus (L.) Link as the type species. It is one of the most economically important and widely distributed fungal genera (Wijayawardene et al. 2017; Houbraken et al. 2020; Zhang et al. 2021). Currently comprising 428 accepted species (Sklenář et al. 2021; Wijayawardene et al. 2022), this taxonomically diverse genus is classified into six subgenera (Aspergillus, Cremei, Circumdati, Fumigati, Nidulantes, and Polypaecilum), 27 sections, and 75 series according to the latest revision (Houbraken et al. 2020; Sklenář et al. 2021). Members of Aspergillus are characterized by their distinctive aspergillum-like conidiophores, consisting of a foot cell, an aseptate stipe, and a terminal vesicle (Houbraken et al. 2020). Most species are terrestrial saprobes, though some occur as aquatic fungi or as opportunistic pathogens (Peterson 2008; Sigler et al. 2010; Varga et al. 2011; Guarro et al. 2012; Samson et al. 2014; Jurjević et al. 2015; Arzanlou et al. 2016; Visagie et al. 2017; Wijayawardene et al. 2017). They are commonly found in soil, plants, and food, with some species posing significant threats as producers of dangerous mycotoxins (aflatoxins and ochratoxins), and others causing aspergillosis in humans, animals, and plants (Peterson 2008; Samson et al. 2014; Al-Bedak and Faysal 2024). Aspergillus is the second most common fungal genus reported from caves, with more than 122 species worldwide, many of which are new cave isolates (Suppl. material 1: table S2). In this study, we describe a new species, Aspergillus dehongensis, and report a new geographical record for A. baeticus A. Nováková & Hubka.

Associated with the rock surface in the cave environment. Asexual morph on PDA: Conidial heads radiate, splitting into three or more columns. Conidiophores uniseriate, light brown with a darker brown upper part. Stipes 150–300 × 4–9 μm (x̄ = 246.88 × 7.44 μm), hyaline to yellow brown, septate. Vesicles 7–28 μm wide, globose to spathulate, smooth. Metulae 8–15 × 5–10 μm (x̄ = 10.18 × 8.38 μm, n = 30), hyaline to pale brown, uni- to biseriate. Phialides 6–13 × 1–3 μm (x̄ = 9.31 × 2.54 μm, n = 30), ampulliform, with minute collarette. Conidia 3–5 × 2–5 μm (x̄ = 3.67 × 3.44 μm, n = 60), globose to subglobose, ornamented with warts or spines, hyaline to yellow brown, arranged in chains. Sexual morph: Undetermined.

Colonies on PDA attaining 20–40 mm after 15 days at room temperature (25 ± 2 °C), colonies at the surface, floccose, wrinkled, with mycelial areas white to cream, greenish-gray to green; reverse white to yellowish gray. Sporulation moderately dense on PDA after 60 days.

Cave sediment (Nováková et al. 2012), rock surface (this study).

Brazil (GBIF 2025), China (this study), India (GBIF 2025), Madagascar (GBIF 2025), Rwanda (GBIF 2025), South Africa (GBIF 2025), Spain (Nováková et al. 2012), Tanzania (GBIF 2025).

CHINA • Yunnan Province, Nujiang Lisu Autonomous Prefecture, Lushui City, Daxingdi Town, Walaya Cave, rock surface, 26.080743°N, 98.848391°E, elev. ca 850.46 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, NJ3D (GMB-W1249), living culture GMBCC2264.

Aspergillus baeticus was introduced by Nováková et al. (2012) from cave sediment in Spain. Our isolate (GMBCC2264) clustered with A. baeticus (NRRL 62501 (type), NRRL 62487, and CMF 2181) within the section Usti with 89% ML and 0.97 BYPP support (Fig. 3). The nucleotide comparison between our strain (GMBCC2264) and A. baeticus (NRRL 62501, type) showed 1.34% (7/524 bp, 3 gaps) differences in BenA, 1.07% (5/469 bp, 1 gap) differences in CAM, and 1.31% (12/914 bp, without gap) differences in rpb2. Morphologically, the new isolate (GMBCC2264) is similar to A. baeticus, with dark brown conidiophores on the upper part and globose to subglobose conidia ornamented with warts or spines (Nováková et al. 2012). Based on phylogenetic and morphological analyses, the new isolate (GMBCC2264) is identified as A. baeticus. This is the first report of A. baeticus isolated from cave environments in China. In addition, this species is known exclusively from cave environments and is reported here for the first time from China.

Named after the type locality, Dehong.

GMB-W1540.

Associated with bamboo sticks, candles, and bat guano in the cave environment. Asexual morph on PDA: Conidial heads radiate, splitting into 3 or more columns. Conidiophores uniseriate, branched at apex. Stipes 200–450 × 3–8 μm (x̄ = 365.67 × 5.43), hyaline, septate. Vesicles 7–28 μm wide, globose to spathulate, smooth-walled. Metulae 6–21 × 3–6 μm (x̄ = 9.78 × 4.38 μm, n = 30), hyaline to pale brown, uni- to biseriate. Phialides 6–16 × 1–3 μm (x̄ = 9.31 × 2.54 μm, n = 30), ampulliform, with minute collarette. Conidia 3–7 × 2–6 μm (x̄ = 4.67 × 3.44 μm, n = 60), broadly ellipsoid to globose, yellowish-green, smooth-walled, hyaline, arranged in chains. Sexual morph: Undetermined.

Colonies on PDA attaining 20–40 mm after 15 days at room temperature (25 ± 2 °C), colonies at surface, floccose, wrinkled, with mycelial areas white to cream, greenish-gray to yellowish-gray; reverse white to brown. Sporulation moderately dense on PDA after 20 days.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Yingjiang County, Nongzhang Town, Xainren Cave, on the candle, 24.524816°N, 97.801635°E, elev. ca 816.49 m, 22 April 2024, Xiangfu Liu and Xuemei Chen, YJ33 (GMB-W1540, holotype), ex-type GMBCC2269, ibid., YJ33C, living culture GZCC 25-0737; ibid., bamboo stick, YJ35C2, living culture GZCC 25-0738; ibid., bat guano, YJ11C, living culture GZCC 25-0736.

In our phylogenetic analyses of ITS, BenA, CaM, and rpb2, Aspergillus dehongensis (GMBCC2269 (type), GZCC 25-0736, GZCC 25-0737, and GZCC 25-0738) formed a distinct lineage sister to A. heterothallicus (CBS 488.65 (type) and CBS 489.65) and A. puniceus (CBS 122.33 and CBS 495.65 (type)) with 100% ML and 1.00 PP support in the Aspergillus section Usti (Fig. 3). However, the nucleotide comparisons between Aspergillus dehongensis (GMBCC2269, type) and A. heterothallicus (CBS 488.65, type) revealed 10.30% (64/427 bp, with 3 gaps) differences in BenA, and 9.97% (109/1011 bp, without gap) base pair differences in rpb2. Aspergillus dehongensis (GMBCC2269, type) and A. puniceus (CBS 495.65, type) revealed 8.41% (36/428 bp, with 6 gaps) differences in BenA, and 6.43% (65/1011 bp, without gaps) base pair differences in rpb2. Morphologically, Aspergillus dehongensis differs from A. heterothallicus in that the latter has aperturate echinulata, smaller conidia (3–7 × 2–6 μm vs. 2.5–4.0 μm in diameter), and irregularly ovoid to elongate, twisted hülle cells compared to unobserved hülle cells of A. dehongensis (Raper and Fennell 1965). Morphologically, Aspergillus dehongensis differs from A. puniceus in that the latter has slightly rough, smaller conidia (3–7 × 2–6 μm vs. 2.5–3.3 μm in diameter), and elongated, crescent-shaped, or irregularly coiled hyphal cells, which aggregate into conspicuous yellow masses (Raper and Fennell 1965). In addition, the PHI test results (Fig. 6A) revealed no significant recombination relationships between A. dehongensis and its phylogenetically related taxa within the dataset, and no strongly supported conflict among individual gene trees (Suppl. material 1: figs S1–S3). Based on the phylogenetic analyses and morphological characteristics, Aspergillus dehongensis is introduced as a new species.

Hypocreaceae was introduced by Lindau (1897) and redefined by Rossman et al. (1999), and is typified by Hypocrea Fr. The sexual morphs of Hypocreaceae are characterized by superficial or immersed, erumpent ascomata, often with papillate apex and rarely bear thick-walled setae, pallid to brightly pigmented, brown, blue or violet peridium, J- asci, often lacking an apical ring. Ascospores are uniseriate, biseriate, or arranged in fascicles, hyaline, yellowish, pinkish to greenish, or occasionally brown, verruculose or longitudinally striate, 1-celled or have one to several septa, occasionally with longitudinal septa (Hyde et al. 2020). In some cases, ascospores disarticulate into part-spores or bud to form conidia within the ascus. The asexual morphs are characterized by hyphomycetous or coelomycetous forms that produce hyaline to brightly colored conidia, although aleurioconidia and ampulloconidia are less commonly observed (Hyde et al. 2020). Some species exhibit a Gliocladium-like asexual morph or possess transversely striate ascospores (Rossman et al. 1999; Perera et al. 2023). Members of the family have been reported as saprobes, endophytes, and pathogens of animals and mushrooms (Dighton 1995; Sung et al. 2007; Back et al. 2010; Yu et al. 2020; Perera et al. 2023). Species of Hypocreaceae are important in agriculture as biocontrol agents and produce microbial natural compounds that are valuable in the pharmaceutical and drug industries (Hyde et al. 2020; Perera et al. 2023). To date, Hypocreaceae comprises 18 accepted genera (Hyde et al. 2024). In previous studies, 67 species belonging to 13 genera of Hypocreaceae have been reported from caves worldwide (Suppl. material 1: table S2).

Trichoderma was established by Persoon (1794), with T. viride as the type and only species retained from the original four (Zhao et al. 2023b; Liu et al. 2025). It is now a large genus with over 500 accepted species, widely distributed in soil, decaying wood, plant leaves, bark, root systems, or other tissues, as well as in caves and even the human body (Zheng et al. 2021; Zhao et al. 2024; Liu et al. 2025). Trichoderma species exhibit saprobic, endophytic, and parasitic lifestyles (Zheng et al. 2021; Zhao et al. 2023b). Members are morphologically characterized by fast-growing colonies, branched conidiophores, flask-shaped phialides, and mostly green conidia (Bissett 1984; Zhao et al. 2023b). Traditional identification relies on morphology; however, species are highly similar. Subsequently, multi-locus approaches have been widely accepted and applied to species identification in Trichoderma (Chaverri and Samuels 2003; Li et al. 2013; Zhu and Zhuang 2015; Zheng et al. 2021; Cao et al. 2022; Zhao et al. 2023b). Recently, Cai and Druzhinina (2021) published an authoritative molecular identification framework for Trichoderma, supported by the International Trichoderma Commission (Cai and Druzhinina 2021; Zhao et al. 2023b, 2024; Liu et al. 2025). At present, more than 500 species have been recognized across 13 clades in Trichoderma (Chaverri et al. 2015; Jaklitsch and Voglmayr 2015; Bustamante et al. 2021; Liu et al. 2025; Xiao et al. 2025). Ecologically and economically, Trichoderma species serve as biocontrol agents, plant growth promoters, and producers of enzymes and antibiotics; however, some strains are pathogenic to mushrooms and animals (Cao et al. 2024; Zeng et al. 2024; Zhao et al. 2024; Liu et al. 2025). Trichoderma is one of the most common genera found in caves, following Aspergillus and Penicillium, with 41 species reported to date from cave environments, including the newly described species reported in Hongsanan et al. (2025) and Liu et al. (2025). According to the standards of Cai and Druzhinina (2021), we introduced seven new species, viz. Trichoderma dehongense, T. hongheense, T. mangshiense, T. neogongcheniae, T. neopyramidale, T. nujiangense, T. yingjiangense, and one new host record in cave habitat.

Named after the type locality, Dehong.

HKAS149895.

Associated with soil in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–5 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the apex. Phialides 5–13 × 3–5 µm (x̄ = 10.47 × 3.41 μm, n = 50), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–5. Supporting cells 5–17 × 2–4 µm (x̄ = 10.09 × 2.97 μm, n = 50). Conidia 4–5 × 3–4 µm (x̄ = 4.01 × 3.79 μm, n = 100), subglobose to globose, or ovoid, light gray to green, smooth-walled. Chlamydospores 4–12 × 4–9 µm (x̄ = 8.28 × 6.23 μm, n = 40), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular with water drops; reverse reddish-brown. The white reproductive mycelium covers the stromatic colonies after 30 days.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, soil, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS14D (HKAS149895, holotype), ex-type GMBCC2301 = GZCC 25-0719, other living culture GZCC 25-0720.

In our phylogenetic analyses of ITS, rpb2, and tef1-α, Trichoderma dehongense (GZCC 25-0719 (ex-type), and GZCC 25-0720) formed a distinct lineage with 83% ML and 0.93 PP support, clustering as the sister taxon to T. afroharzianum and T. mangshiense in the harzianum clade (Fig. 7). Moreover, nucleotide comparisons between Trichoderma dehongense (GMBCC2301, ex-type) and T. afroharzianum (CBS 124620, ex-type) revealed 1.04% (9/836 bp, without gap) base pair differences in rpb2, and 6.04% (20/331 bp, with 4 gaps) differences in tef1-α. Trichoderma dehongense (GMBCC2301, ex-type) and T. mangshiense (GMBCC2300, ex-type) showed 2.59% (22/849 bp, 2 gaps) base pair differences in rpb2, and 4.02% (14/348 bp, with 3 gaps) differences in tef1-α. Morphologically, T. dehongense is different from T. afroharzianum by having more whorls (2–5 vs 1–3), longer phialides (5–13 × 3–5 µm vs 5.2–10.2 × 2.5–3.5 µm) on terminal branches, larger conidia (4–5 × 3–4 µm vs 2.7–3.5 × 2.5–3.2 μm), and subglobose to globose, thick-walled chlamydospores, compared to rare chlamydospores of T. afroharzianum (Chaverri et al. 2015). Trichoderma dehongense is different from T. mangshiense by more whorls (2–5 vs 2–3), shorter phialides (5–13 × 3–5 µm vs 7–15 × 3–5 µm) on terminal branches, and longer chlamydospores (4–12 × 4–9 µm vs 4–9.5 × 4–9 µm). Trichoderma afroharzianum was first introduced in a phylogenetic study of T. harzianum by Druzhinina et al. (2010) but was not validly published because it lacked a formal

Chaverri et al. (2015) later reintroduced and validly published the species under the name T. afroharzianum, providing detailed descriptions. In addition, the PHI test results (Fig. 6B1) revealed no significant recombination relationships between T. dehongense and its phylogenetically related taxa within the dataset. Based on phylogenetic analyses and morphological characteristics, Trichoderma dehongense is herein described as a new species.

Named after the type locality, Honghe.

HKAS149896.

Associated with the air in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–4 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 5–13 × 2–5 µm (x̄ = 8.26 × 2.98 μm, n = 60), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–4. Supporting cells 5–11 × 2–4 µm (x̄ = 8.17 × 2.81 μm, n = 50). Conidia 2–5 × 2–4 µm (x̄ = 3.59 × 3.24 μm, n = 60), subglobose to globose, or ovoid, green, smooth-walled. Chlamydospores 5–11 × 3–9 µm (x̄ = 8.15 × 6.66 μm, n = 10), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to yellow or green, sparse aerial mycelium on the surface, regularly circular, green conidiation occurs over time; reverse is green in the middle and white on the edges.

CHINA • Yunnan Province, Honghe Hani and Yi Autonomous Prefecture, Jianshui County, Miandian Town, Swallow Cave, 23.63684309°N, 103.05537745°E, elev. ca 1864.03 m, air, 24 December 2020, Xiangfu Liu, A7 (HKAS149896, holotype), ex-type GMBCC2297; ibid., A7B, living culture GMBCC2298.

Phylogenetic analyses revealed that Trichoderma hongheense (GMBCC2297 (ex-type), and GMBCC2298) forms a distinct lineage with 100% ML and 1.00 PP support, and is closely related to T. hipposidericola Karun., Tibpromma & X.F. Liu, and T. atroviride P. Karst. in the atroviride clade (Fig. 7). The nucleotide comparisons between Trichoderma hongheense and T. hipposidericola (KUMCC 21-0801, ex-type) revealed 1.51% (13/869 bp, without gap) differences in rpb2 and 2.19% (16/731 bp, with 5 gaps) differences in tef1-α. Trichoderma hongheense and T. atroviride (CBS 142.95, ex-type) revealed 2.30% (20/869 bp, without gap) differences in rpb2, and 2.05% (15/731 bp, with 5 gaps) differences in tef1-α. Morphologically, T. hongheense differs from T. hipposidericola in having more whorls of phialides (2–4 vs 2–5) on terminal branches and lacking chlamydospores (Liu et al. 2023). In addition, the PHI test results (Fig. 6B2) revealed no significant recombination relationships between T. hongheense and its phylogenetically related taxa within the dataset. Based on the multi-gene phylogenetic analyses and morphological characteristics, Trichoderma hongheense is described as a new species.

Named after the type locality, Mangshi.

GMB-W1539.

Associated with soil in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–4 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 7–15 × 3–5 µm (x̄ = 10.27 × 3.43 μm, n = 50), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–3. Supporting cells 5–17 × 2–4 µm (x̄ = 10.09 × 2.97 μm, n = 50). Conidia 4–5 × 3–4 µm (x̄ = 4.01 × 3.79 μm, n = 100), subglobose to globose or ovoid, light gray to green, smooth-walled. Chlamydospores 4–9.5 × 4–9 µm (x̄ = 6.28 × 5.92 μm, n = 40), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular with water drops; reverse reddish-brown. The white reproductive mycelium covers the stromatic colonies after 30 days.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, soil, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS14A (GMB-W1539, holotype), ex-type GMBCC2300, other living culture GZCC 25-0718.

Phylogenetic analysis of combined ITS, tef1-α, and rpb2 sequence data, Trichoderma mangshiense (GMBCC2300 (ex-type), and GZCC 25-0718) forms a lineage separated from T. dehongense and T. afroharzianum in the harzianum clade (Fig. 7). Moreover, nucleotide comparisons between T. mangshiense (GMBCC2300, ex-type) and T. afroharzianum (CBS 124620, ex-type) revealed 2.91% (30/886 bp, with 2 gaps) base pair differences in rpb2, and 5.41% (18/333 bp, with 3 gaps) differences in tef1-α. Morphologically, T. mangshiense differs from T. afroharzianum by having larger phialides (7–15 × 3–5 µm vs. 5.2–10.2 × 2.5–3.5 µm), smaller conidia (4–5 × 3–4 µm vs 2.7–3.5 × 2.5–3.2 µm), and rarely observed chlamydospores (Chaverri et al. 2015). Morphological differences between T. mangshiense and T. dehongense were listed under the notes of T. dehongense. In addition, the PHI test results (Fig. 6B1) revealed no significant recombination relationships between T. mangshiense and its phylogenetically related taxa within the dataset. Based on phylogenetic analyses and morphological characteristics, Trichoderma mangshiense is introduced as a new species.

Named after its phylogenetic closeness to T. gongcheniae.

GMB-W1538.

Associated with soil in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–4 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 3–11 × 3–8 µm (x̄ = 6.28 × 4.51 μm, n = 50), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–3. Supporting cells (3.8–) 6–14 (–21) × 2–4.5 µm (x̄ = 9.08 × 3.36 μm, n = 50). Conidia 3–4 × 2.5–4 µm (x̄ = 3.08 × 2.98 μm, n = 50), subglobose to globose or ovoid, green, smooth-walled. Chlamydospores 6–9 (–11) × 5–8 µm (x̄ = 7.87 × 6.07 μm, n = 15), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular, green conidiation occurs over time; reverse is green in the middle and white on the edges.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, feather, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS17B (GMB-W1538, holotype), ex-type GZCC 25-0721, other living cultures GMBCC2302, GZCC 25-0722.

Trichoderma neogongcheniae (GZCC 25-0721 (ex-type), GZCC 25-0722, and GMBCC2302) is phylogenetically closely related to T. gongcheniae (Fig. 7). The morphology of these two species is similar in phialides, supporting cells, and conidia. However, the size of the phialides in T. gongcheniae (5.6–8.4 × 2.8–3.7 μm) is smaller than that in T. neogongcheniae (3–11 × 3–8 μm). Additionally, the conidia of T. neogongcheniae (3–4 × 2.5–4 μm) are larger than those of T. gongcheniae (2.8–3.3 × 2.7–3.1 μm). In terms of chlamydospores, T. neogongcheniae has bigger chlamydospores (6–9 × 5–8 μm) compared to T. gongcheniae (5.2–6.8 × 4.8–6.3 μm) (Zhao et al. 2024). Meanwhile, nucleotide comparisons between Trichoderma neogongcheniae (GZCC 25-0721, ex-type) and T. gongcheniae (GDMCC 3.1011, ex-type) revealed 2.68% (24/897 bp) base pair differences in rpb2. In addition, the PHI test results (Fig. 6B3) revealed no significant recombination relationships between T. neogongcheniae and its phylogenetically related taxa within the dataset. Therefore, we introduce Trichoderma neogongcheniae as a new species.

Named after its phylogenetically closeness to Trichoderma pseudopyramidale and morphological similarity to Trichoderma pyramidale.

GMB-W1534.

Associated with rock surface in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–8 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 5–15 × 2–4 µm (x̄ = 8.82 × 2.79 μm, n = 60), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 1–3. Supporting cells 5–13 × 2–4 µm (x̄ = 7.91 × 2.63 μm, n = 50). Conidia 2–5 × 2–4 µm (x̄ = 3.44 × 2.95 μm, n = 90), subglobose to globose, or ovoid, green, smooth-walled. Chlamydospores 4–6 × 3–5 µm (x̄ = 5.14 × 4.34 μm, n = 10), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular, green conidiation occurs over time, reverse is green in the middle and white on the edges.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, rock surface, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS32C (GMB-W1534, holotype), ex-type GMBCC2303, ibid., MS32D (GMB-W1535, isotype), ex-isotype GZCC 25-0729.

Phylogenetic analyses revealed that Trichoderma neopyramidale (GMBCC2303 (ex-type), and GZCC 25-0729) forms a distinct lineage and is closely related to T. pseudopyramidale and T. hydei X.F. Liu, Karun. & Tibpromma (Fig. 7). Trichoderma neopyramidale (GMBCC2303, ex-type) and T. pseudopyramidale (GMBCC2300, ex-type) revealed 2.52% (21/833 bp, 2gaps) base pair differences in rpb2, and 6.04% (35/579 bp, without gaps) differences in tef1-α. Trichoderma neopyramidale (GMBCC2303, ex-type) and T. hydei (GMBCC1136, ex-type) revealed 1.77% (16/904 bp, without gap) base pair differences in rpb2, and 2.92% (23/787 bp, without gap) differences in tef1-α. Morphologically, T. neopyramidale differs from T. pseudopyramidale by the latter having smaller phialides (5–15 × 2–4 µm vs. 5.3–8.6 × 2.2–2.9 µm), smaller supporting cells (5–13 × 2–4 µm vs. 3.3–7.1 × 1.9–2.8 µm), and larger conidia (2–5 × 2–4 µm vs. 2.1–2.9 × 2.3–2.9 µm) (Rodríguez et al. 2021). Trichoderma neopyramidale differs from T. hydei by the latter having white, gray to green culture, more (1–3 whorls vs. 2–3 whorls) and slightly longer (5–15 × 2–4 µm vs. 6–17 × 2–4 µm) phialides, bigger supporting cells (5–13 × 2–4 µm vs. 6–15 × 2–4 µm), and smaller conidia (2–5 × 2–4 µm vs. 2–4 × 2–3 µm) (Hongsanan et al. 2025). Trichoderma neopyramidale and T. pyramidale share several characteristics in common, such as pyramidal conidiophores with 2−4 whorls phialides. Trichoderma pyramidale has smalller phialides ((4.0−)5.5−11.5(−17.5) × (2.5−)2.8−3.7(−4.5)) and conidia ((2.8−) 3.2− 4.0(−4.7) × (2.5−) 3.0−3.5(−4.0) μm), and uncommon chlamydospores (Chaverri et al. 2015). In addition, the PHI test results (Fig. 6B4) revealed no significant recombination relationships between T. neopyramidale and its phylogenetically related taxa within the dataset. Based on phylogenetic analyses and morphological characteristics, Trichoderma neopyramidale is introduced as a new species.

Named after the type locality, Nujiang.

GMB-W1546.

Associated with rock surface in the cave environment. Asexual morph on PDA: Vegetative hyphae 2–6 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 5–11 × 2–5 µm (x̄ = 7.94 × 3.32 μm, n = 50), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–3. Supporting cells 5–13 (–18) × 2–4 µm (x̄ = 9.11 × 2.92 μm, n = 50). Conidia 3–4 × 2–4 µm (x̄ = 3.28 × 3.21 μm, n = 50), subglobose to globose, or ovoid, green, smooth-walled. Chlamydospores 5–8 (–14) × 5–8 (–9) µm (x̄ = 7.13 × 6.76 μm, n = 20), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular, green conidiation occurs over time; reverse is green in the middle and white on the edges.

CHINA • Yunnan Province, Nujiang Lisu Autonomous Prefecture, Lushui City, Daxingdi Town, Walaya Cave, rock surface, 26.080743°N, 98.848391°E, elev. ca 850.46 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, NJ49A (GMB-W1546, holotype), ex-type GMBCC2304, other living culture GZCC 25-0733.

Phylogenetic analyses revealed that Trichoderma nujiangense (GMBCC2304, ex-type) is closely related to T. cavernicola (GMBCC1103 (ex-type) and GMBCC1105) in Fig. 7. Morphologically, Trichoderma cavernicola differs from T. nujiangense by having more (2–4(–5) whorls vs. 2–3 whorls) and longer (6–17 × 2–4 µm vs. 5–11 × 2–5 µm) phialides on terminal branches, bigger supporting cells (4–22 × 2–8 µm vs. 5–13 (–18) × 2–4 µm), and smaller conidia (4–7 × 2–5 µm vs. 3–4 × 2–4 µm) (Liu et al. 2025). The nucleotide comparisons between T. nujiangense (GMBCC2304, ex-type) and T. cavernicola (GMBCC1103, ex-type) revealed 2.35% (15/638 bp, with 2 gaps) base pair differences in ITS, 4.37% (39/892 bp, without gaps) differences in rpb2, and 0.38% (3/787 bp, without gaps) differences in tef1-α. In addition, the PHI test results (Fig. 6B3) revealed no significant recombination relationships between T. nujiangense and its phylogenetically related taxa within the dataset. Therefore, we introduce Trichoderma nujiangense as a new species.

Associated with petroleum wax in the cave environment. Asexual morph on PDA: Vegetative hyphae septate, branched, hyaline, smooth-walled, 1–5 μm wide. Conidiophores pyramidal with verticillate, paired lateral branches. Phialides 3–12 × 1–4 µm (x̄ = 6.83 × 2.64 μm, n = 30), generally formed on terminal branches, ampulliform, in divergent whorls of 1–4. Supporting cells 3–16 × 1–4 µm (x̄ = 8.99 × 2.97 μm, n = 30). Conidia 3–6 × 2–5 µm (x̄ = 4.26 × 3.55 μm, n = 60) globose, ellipsoidal to ovoid, green, smooth-walled. Chlamydospores not observed. Sexual morph: Undetermined.

Colonies on PDA attaining 50–60 mm after 7 days at room temperature (25 ± 2 °C), white, pale yellow to green, sparse aerial mycelium on the surface, reverse white to pale yellow. The white reproductive mycelium covers the stromatic colonies after 25 days. Odor absent.

Feet surface of living bats (Liu et al. 2023), petroleum wax (this study), and soil (Zheng et al. 2021).

China (Zheng et al. 2021; Liu et al. 2023; this study).

CHINA • Yunnan Province, Nujiang Lisu Autonomous Prefecture, Lushui City, Daxingdi Town, Walaya Cave, petroleum wax, 26.080743°N, 98.848391°E, elev. ca 850.46 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, NJ5C (GMB-W1250), living culture GMBCC2473.

Trichoderma obovatum was erected by Zheng et al. (2021) and was isolated from soil in Yunnan, China. Our isolate (GMBCC2473) clustered with T. obovatum (YMF 1.06211 (ex-type) and YMF 1.06212) within the Atroviride clade (Fig. 7). The nucleotide comparison between our strain (GMBCC2473) and T. obovatum (YMF 1.06211, ex-type) showed 0.35% (2/572 bp, 2 gaps) base pair differences in ITS, 0.44% (4/904 bp, 1 gap) differences in rpb2, and no (0/712 bp, without gap) differences in tef1-α. Morphologically, the new isolate (GMBCC2473) is similar to T. obovatum in having curved to sinuous phialides, globose to oval or obovate conidia of slightly different sizes, and the absence of chlamydospores (Zheng et al. 2021; Liu et al. 2023). Based on combined phylogeny and morphological evidence, our strains were identified as T. obovatum. This species has previously been isolated from soil and bat surfaces in China (Zheng et al. 2021; Liu et al. 2023); however, this represents the first record of T. obovatum from a cave environment.

Named after the type locality, Yingjiang.

HKAS149894.

Associated with bat guano in a cave environment. Asexual morph on PDA: Vegetative hyphae 2–8 μm wide, septate, branched, hyaline, smooth-walled. Conidiophores pyramidal with verticillate, comprising a recognizable main axis, pairs or solitary branches, longer branches near the base, and short branches or solitary phialides arising near the tip. Phialides 6–12 × 2–4 µm (x̄ = 7.99 × 2.83 μm, n = 100), ampulliform to lageniform, generally formed on terminal branches in divergent whorls of 2–4. Supporting cells 4–14 × 2–4 µm (x̄ = 7.64 × 2.61 μm, n = 40). Conidia 2–5 × 2–4 µm (x̄ = 3.17 × 2.72 μm, n = 100), subglobose to globose, or ovoid, green, smooth-walled. Chlamydospores 4–9 × 3–7 µm (x̄ = 6.21 × 5.14 μm, n = 10), single or in clusters, terminal and intercalary, variable in shape, mostly subglobose to globose, thick-walled, hyaline, granulate. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), white, gray to green, sparse aerial mycelium on the surface, regularly circular, green conidiation occurs over time; reverse is green in the middle and white on the edges.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, bat guano, 24.232220°N, 98.426103°E, elev. ca 1530.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS19B (HKAS149894, holotype), ex-type GZCC 25-0723, other living culture GZCC 25-0724.

Trichoderma yingjiangense (GZCC 25-0723, ex-type) is phylogenetically closely related to T. graminis C.L. Zhang and T. bannaense Kai Chen & W.Y. Zhuang in the harzianum clade (Fig. 7). However, Trichoderma yingjiangense is different from T. graminis by the latter having 2–5 whorls and of shorter phialides (6–12 × 2–4 µm vs 5.2–8.1 × 2.7–3.9 µm) on terminal branches, slightly smaller conidia (2–5 × 2–4 µm vs 3.1–4.2 × 2.9–3.9 µm), and smaller chlamydospores (4–9 × 3–7 µm vs 5.6–7.6 × (5.1–)5.3–7.1 μm) (Zhao et al. 2024). Furthermore, T. graminis was isolated as an endophyte from living leaf sheaths of Oplismenus undulatifolius (Zhao et al. 2024). Trichoderma yingjiangense differs from T. bannaense in that the latter has more whorls (2–4 vs 2–5) of shorter phialides (6–12 × 2–4 µm vs 7.5–10 × 2.6–3.6 μm) on terminal branches, smaller conidia (2–5 × 2–4 µm vs 2.5–3.6 × 2.5–3.1 μm), and lacks chlamydospores (Chen and Zhuang 2017). The nucleotide comparisons between T. yingjiangense (GZCC 25-0723, ex-type) and T. graminis (GDMCC 3.1013, ex-type) showed 4.89% (44/900 bp, without gap) base pair differences in rpb2 and 2.03% (16/790 bp, with 6 gaps) differences in tef1-α. Nucleotide comparison between T. yingjiangense (GZCC 25-0723, ex-type) and T. bannaense (HMAS 248840, ex-type) showed 2.55% (15/589 bp, without gap) base pair differences in ITS, 3.43% (26/757 bp, without gap) base pair differences in rpb2, and 2.03% (16/790 bp, with 6 gaps) differences in tef1-α. In addition, the PHI test results (Fig. 6B5) revealed no significant recombination relationships between T. yingjiangense and its phylogenetically related taxa within the dataset. Based on the phylogenetic analyses and morphological characteristics, Trichoderma yingjiangense is introduced as a new species.

Nectriaceae was established by Tulasne and Tulasne (1865) for the genus Nectria (Fr.) Fr. and was later accepted as a distinct family by Rossman et al. (1999) based on morphological characteristics. The sexual morphs of Nectriaceae are characterized by uniloculate ascomata that are yellow, orange, red, purple, or brown, and often exhibit color changes when treated with 3% potassium hydroxide (KOH) or 100% lactic acid (LA). These ascomata are globose to long-fusiform, with ellipsoid forms also occurring, and produce non-disarticulating ascospores (Rossman et al. 1999; Lombard et al. 2015; Zeng and Zhuang 2022; Perera et al. 2023). The asexual morphs are characterized by phialidic conidiogenous cells, producing aseptate to multi-septate conidia, which may be constricted at the septa or not, with or without distinct abscission scars; chlamydospores may be present or absent (Maharachchikumbura et al. 2016; Perera et al. 2023). Species of Nectriaceae have been reported as saprobes, pathogens, or endophytes, and have been isolated from a wide range of substrates, including living and decaying wood material, soil, freshwater habitats, insects, fruiting bodies of other fungi, and various other materials across tropical and subtropical regions worldwide (Rossman et al. 1999; Morrison et al. 2021; Zeng and Zhuang 2022; Rissi et al. 2024). Species of Nectriaceae are of considerable importance in biotechnological, medicinal, commercial, pharmaceutical, and industrial applications (Abdel-Azeem et al. 2019; Peitl et al. 2020; Cleveland et al. 2021; Ibrahim et al. 2021; Perera et al. 2023). To date, 38 species belonging to eight genera of Nectriaceae have been reported from caves worldwide (Suppl. material 1: table S2).

Cosmospora was introduced by Rabenhorst (1862) and typified by C. coccinea Rabenh. The genus is distributed across temperate and tropical regions (Gräfenhan et al. 2011), with approximately 87 epithets currently recorded in Index Fungorum (http://www.indexfungorum.org; accessed date 14 July 2026). Species of Cosmospora are mainly reported as saprobic or endophytic and are found in warm-temperate to tropical regions (Peintner et al. 2002; Gräfenhan et al. 2011; Herrera et al. 2015; Zeng and Zhuang 2016; Bao et al. 2023). The sexual morph of Cosmospora is characterized by orange-red to bright-red perithecia that turn dark red in KOH. The ascomata initially contain hyaline asci that become yellow-brown to reddish-brown and eventually develop into tuberculate structures at maturity. Ascospores are multi-septate and verrucose or tuberculate. The asexual morph is characterized by monophialidic phialides and aseptate conidia that are ellipsoidal, oblong, clavate, or allantoid (Lombard et al. 2015; Zeng and Zhuang 2016; Bao et al. 2023). Previous studies have reported more than five Cosmospora species from cave environments (Suppl. material 1: table S2). In this study, we introduce Cosmospora cavernicola sp. nov., isolated from the rock surface in a cave.

“Caverna” means cave in Latin, and “cola” means dweller or inhabitant, hence the creation of the epithet.

GMB-W1537.

Associated with rock surface in the cave environment. Asexual morph on PDA: Mycelium consisting of branched, septate, hyaline to subhyaline or yellowish-orange in mass, smooth hyphae, 1.1–3.2 µm diam., with occasional anastomoses. Conidiophores arising laterally or terminally from somatic hyphae, verticillate, up to 230 μm long, branched, hyaline, smooth and thin-walled, bearing terminal and lateral conidiogenous cells, mostly bearing whorls of 2 conidiogenous cells. Conidiogenous cells 21–92 µm long, 1–4 µm wide at the base, enteroblastic, monophialidic, subulate or subcylindrical, terminal or lateral, discrete, straight, septate, tapering towards the apex, smooth-walled, hyaline, without a distinct periclinal thickening, 0.6–2.2 µm wide at the apex. Microconidia 3–5 × 1–3 μm (x̄ = 3.85 × 2.97 μm, n = 90), ellipsoidal, cylindrical, slightly allantoid or obovoid to clavate, sometimes flattened on one side, hyaline, smooth- and thin-walled, aseptate. Chlamydospores not observed. Sexual morph: Undetermined.

Colonies on PDA attaining 50 mm after 30 days at room temperature (25 ± 2 °C), creamy at the center, becoming yellow toward the outer part of the colony with a white margin, with a fimbriate margin; reverse brown and dark yellow with a light-yellow margin. Odor absent.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, rock surface, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS25D (GMB-W1537, holotype), ex-type GZCC 25-0728; ibid., MS25B2 (GMB-W1243, isotype), ex-isotype GMBCC2272, ibid., MS25C (GMB-W1244, isotype), ex-isotype GZCC 25-0727.

In the multi-gene phylogenetic analyses of the combined ITS, LSU, rpb2, and tub2 regions, Cosmospora cavernicola (GZCC 25-0728 (ex-type), GMBCC2272 and GZCC 25-0727) formed a distinct lineage to Co. khandalensis (Thirum. & Sukapure) Gräfenhan & Seifert with 100% ML and 1.00 PP support (Fig. 16). However, the nucleotide comparisons between Co. cavernicola (GZCC 25-0728, ex-type) and Co. khandalensis (IMI 112790, ex-type) showed 1.61% (18/544 bp, 2 gaps) base pair differences in ITS, 0.13% (1/788 bp, without gap) differences in LSU, 2.20% (7/318 bp, without gap) differences in tub2, and 0.81% (7/867 bp, without gap) differences in rpb2. Morphologically, Cosmospora cavernicola can be easily distinguished from Co. khandalensis by its verticillate conidiophores and longer, septate conidiogenous cells (21–92 µm vs. 34–64 µm) (Herrera et al. 2015). In addition, the PHI test results (Fig. 6C) revealed no significant recombination relationships between C. cavernicola and its phylogenetically related taxa within the dataset. This fungus is, therefore, introduced as a new species of Cosmospora.

“Caverna” means cave, and “cola” means dweller or inhabitant in Latin; thus, “cavernicola” refers to the cave habitat of the holotype.

GMB-W1482.

Associated with rock surface in the cave. Asexual morph on PDA: Vegetative hyphae 1–2.5 μm wide, branched, hyaline, septate, smooth-walled. Conidiophores arising from substrate, mycelium simple to indistinctive. Conidiogenous cells 2–7 × 1–4 µm (x̄ = 4.648 × 2.34 μm, n = 30), hyaline, cylindrical to ampulliform, reduced to short swollen supporting cells, annellidic, septate, smooth, thin-walled, born singly, often laterally and rarely apically on aerial hyphae. Conidia 3–6 × 2–5 µm (x̄ = 4.62 × 3.77 μm, n = 60), globose to pyriform, with a truncate base, hyaline to pale brown, smooth and thin-walled, aseptate, arranged in chains, with a disjuncture between adjacent conidia. Chlamydospores not observed. Sexual morph: Undetermined.

Colonies on PDA are 35–45 mm diam in 14 d at room temperature (25 ± 2 °C), flat, felty to floccose, and pale gray near the center, with a pale brown irregular margin; the reverse is pale brown near the center and white near the margin.

CHINA • Yunnan Province, Honghe Hani and Yi Autonomous Prefecture, Jianshui County, Miandian Town, Swallow Cave, rock surfaces, 26.080743°N, 98.848391°E, elev. ca 1343 m, 24 December 2020, Xiangfu Liu, YZD14C (GMB-W1482, holotype), ex-type GMBCC2294; ibid., YZD14C2 (GMB-W1258, isotype), ex-isotype GMBCC2476.

The new species was identified as a member of Pithoascus based on combined morphological and phylogenetic evidence. In our phylogenetic analyses of combined ITS, LSU, tef1-α, and tub2, Pithoascus cavernicola (GMBCC2294 and GMBCC2476) formed a distinct lineage with 85% ML and 1.00 PP support (Fig. 18). Pairwise nucleotide comparisons between P. cavernicola (GMBCC2294, ex-type) and closely related species revealed significant sequence divergence. Pithoascus cavernicola differs from P. ater (CBS 400.34, ex-type) by 6.72% (32/476 bp, 13 gaps) base pair differences in ITS, 2.03% (11/543 bp, 1 gap) differences in LSU, and 5.57% (53/952 bp, 4 gaps) differences in tef1-α. Comparisons with P. exsertus (CBS 81 9.70, ex-type) showed 9.57% (51/533 bp, 18 gaps) base pair differences in ITS, 1.49% (8/536 bp, without gap) differences in LSU, and 7.18% (69/961 bp, 5 gaps) differences in tef1-α. Pithoascus cavernicola (GMBCC2294, ex-type) differs from P. intermedius (CBS 217.32, ex-type) by 5.44% (29/533 bp, 19 gaps) base pair differences in ITS, 1.50% (8/535 bp, 1 gap) differences in LSU, and 6.26% (60/959 bp, 4 gaps) differences in tef1-α.

Sequence comparisons with P. kurdistanensis (IRAN 4653C, type) showed 5.22% (50/958 bp, 4 gaps) base pair differences in tef1-α. Pithoascus cavernicola (GMBCC2294, ex-type) differs from P. lunatus (CBS 103.85, ex-type) by 6.83% (20/293 bp, 7 gaps) base pair differences in ITS, 1.32% (11/831 bp, 1 gap) differences in LSU, and 6.36% (64/1006 bp, 6 gaps) differences in tef1-α. Pithoascus cavernicola (GMBCC2294, ex-type) differs from P. persicus (IRAN 3309C, ex-type) by 9.19% (49/533 bp, 34 gaps) base pair differences in ITS, 2.46% (20/812 bp, 2 gaps) differences in LSU, and 6.46% (48/743 bp, 3 gaps) differences in tef1-α. Pithoascus cavernicola (GMBCC2294, ex-type) and P. stoveri (CBS 176.71, ex-type) sequences showed 11.44% (61/533 bp, 35 gaps) base pair differences in ITS, 2.43% (13/536 bp, 3 gaps) base pair differences in LSU, and 5.31% (51/961 bp, 4 gaps) base pair differences in tef1-α.

Morphologically, P. cavernicola differs from P. exsertus and P. lunatus in the characteristics of its asexual morph. It differs from P. ater by the latter producing solitary, globose, and larger conidia (3–6 × 2–5 µm vs. 4–9 × 4.5–8.5 µm) (Sandoval-Denis et al. 2016). Pithoascus cavernicola differs from P. intermedius in that the latter has solitary, hyaline, and larger (3–6 × 2–5 µm vs. 4–8 × 4.5–7.5 µm) conidia, whereas those of P. cavernicola are arranged in conidia (Mohammadi et al. 2024). Pithoascus cavernicola differs from P. kurdistanensis by the latter having cylindrical to ampulliform conidiogenous cells, rarely reduced to short swollen supporting cells, slightly smaller (3–6 × 2–5 µm vs 3.5–5 × 3.5–4 µm) conidia, and setae, compared to conidia of P. cavernicola that are arranged in chains (Mohammadi et al. 2024). Pithoascus cavernicola differs from P. persicus in that the latter has annellidic, short, ampulliform conidiogenous cells, and smaller (3–6 × 2–5 µm vs. 2.5 µm in diameter) globose conidia (Tazik et al. 2020). Pithoascus cavernicola differs from P. stoveri in that the latter has annellidic, short, ampulliform conidiogenous cells, and hyaline, obovate to pyriform conidia that are larger (3–6 × 2–5 µm vs 5–8 × 3–4 µm) (Sandoval-Denis et al. 2016; Mohammadi et al. 2024). In addition, the PHI test results (Fig. 6D) revealed no significant recombination relationships between P. cavernicola and its phylogenetically related taxa within the dataset. The phylogenetic and morphological results indicate that P. cavernicola is a distinct new species.

Named after its morphological similarity to Achaetomiella.

Neoachaetomiella cavernicola X.F. Liu, Karun., Tibpromma & K.D. Hyde.

Associated with rock surfaces of caves. Sexual morph: Ascomata superficial, subglobose or ovate, with brown walls of textura angularis in surface view, ostiolate. Terminal hairs flexuous, undulate, or spirally coiled. Lateral hairs straight, flexuous. Asci clavate or fusiform, stalked, evanescent. Ascospores biseriate or irregularly arranged, olivaceous to olivaceous brown at maturity, limoniform or fusiform, bilaterally flattened, with an apical germ pore, usually more than 7 μm and less than 15 μm in length. Asexual morph: not observed.

Based on phylogenetic analyses, our strains formed an independent lineage well separated from Achaetomiella Arx and Collariella X. Wei Wang, Samson & Crous (Fig. 20). Neoachaetomiella can be distinguished from Achaetomiella and Collariella by its olivaceous to olivaceous-brown, limoniform to fusiform, bilaterally flattened ascospores, which possess an apical germ pore and measure typically 7–15 μm in length (Wang et al. 2016a, 2022). Achaetomiella consists of two species (Achaetomiella gracilis and A. virescens) with sexual morphs, producing brown, ellipsoidal or fusiform ascospores, usually longer than 9 μm compared to more than 7 μm and less than 15 μm of Neoachaetomiella (Wang et al. 2022). In contrast, Collariella, which includes 12 species with sexual morphs, has broadly limoniform to quadrangular, olivaceous-brown ascospores, typically less than 7.5 μm in length, occasionally up to 10.5 μm, compared with more than 7 μm and less than 15 μm in Neoachaetomiella (Wang et al. 2022). Megablast searches of GenBank revealed that the closest hits to Neoachaetomiella ITS sequences showed 93.79% similarity; LSU sequences showed 99.50% similarity; rpb2 sequences had 84.27% similarity, and tub2 sequences showed 81.78% similarity, supporting its recognition as a distinct genus.

“Caverna” means cave in Latin, and “cola” means dweller or inhabitant, hence the creation of the epithet.

GMB-W1532.

Associated with rock surface in the cave. Sexual morph on PDA: Ascomata 300–600 × 150–350 μm, usually covered by thick aerial hyphae, olivaceous or umber to dark-brick in reflected light owing to ascomatal hairs, obovate or ovate, ostiolate. Terminal hairs errucose, brown, erect to flexuous or slightly undulate, usually unbranched, 2–6 μm near the base, tapering towards the tips. Lateral hairs are similar. Asci 38–80 × 6–18 μm (x̄ = 56.06 × 12.34, n = 60), 8-spored, fasciculate, fusiform or clavate, evanescent. Ascospores 7–13 × 5–9 μm (x̄ = 9.58 × 7.09, n = 150), biseriate, hyaline to olivaceous brown at maturity, limoniform, some globose, verruculose, thick-walled, biapiculate, bilaterally flattened, sometimes with distinct oil droplets and an apical germ pore. Asexual morph: Undetermined.

Colony on PDA attaining 50 mm after 15 days at room temperature (25 ± 2 °C), folded, white to light yellow, margin undulate to erose to fimbriate. Ascomata arise from the colony, yellowish-brown after 50 days. Odor absent.

CHINA • Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Mang City, Mengga Town, Sanxian Cave, on rock surface, 24.232220°N, 98.426103°E, elev. ca 1528.89 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, MS41D (GMB-W1532, holotype), ex-type GMBCC2270, ibid., MS41D2 (GMB-W1533, isotype), ex-isotype GZCC 25-0730.

In the multi-gene phylogenetic analyses of the combined ITS, LSU, rpb2, and tub2 regions, Neoachaetomiella cavernicola (GMBCC2270 and GZCC 25-0730) formed a distinct lineage, well separated from Achaetomiella and Collariella (Fig. 20). Morphologically, N. cavernicola is similar to these genera but is distinguished by olivaceous to olivaceous-brown, limoniform to fusiform, bilaterally flattened ascospores, with an apical germ pore, typically 7–15 μm in length. In contrast, Achaetomiella is characterized by simple ascomatal hairs evenly distributed over the ascoma, and ellipsoidal or fusiform ascospores with a length usually more than 9 μm, and Collariella is characterized by broadly limoniform to quadrangular, bilaterally flattened ascospores with an apical germ pore, with a length generally less than 7.5 μm (Wang et al. 2022). In addition, the PHI test results (Fig. 6E) revealed no significant recombination relationships between N. cavernicola and its phylogenetically related taxa within the dataset. Herein, we introduce Neoachaetomiella cavernicola as a new species based on both morphological comparison and phylogenetic analyses.

Chaetomium was introduced by Kunze and Schmidt (1817), typified by Chaetomium globosum Kunze (Wijayawardene et al. 2017), is currently the largest genus in Chaetomiaceae (Hyde et al. 2020), comprising 359 accepted species, as outlined in the latest update by Wijayawardene et al. (2022), and 449 epithets are recorded in Index Fungorum (http://www.indexfungorum.org; accessed date 14 July 2026). Members of this genus live in plants, soil, straw, and dung as saprobic fungi worldwide, and some have potential as biological control agents (Ahmed et al. 2012; Mungai et al. 2012; Maharachchikumbura et al. 2015, 2016; Réblová et al. 2016; Wang et al. 2016a, 2016b; Wijayawardene et al. 2017). The genus is characterized by globose, ellipsoid to ovate or obovate, ostiolate ascomata with hypha-like, flexuous ascomatal hairs, limoniform to globose, bilaterally flattened ascospores, and asexual morphs that are hyphomycetous and acremonium-like (Wang et al. 2016a; Hyde et al. 2020). To date, over nine Chaetomium species have been reported from various cave substrates (Suppl. material 1: table S2). In this study, we report a new substrate record of Chaetomium species from caves in Yunnan Province, China.

Associated with decaying twigs in the cave environment. Sexual morph on PDA: Ascomata 200–400 × 150–350 μm, obovate or ovate, ostiolate, usually covered by thick aerial hyphae, olivaceous or umber to dark-brick in reflected light owing to ascomatal hairs. Terminal hairs verrucose, brown, erect to flexuous or slightly undulate, usually unbranched, 2–4 μm near the base, tapering towards the tips. Lateral hairs similar. Asci 8-spored, fasciculate, fusiform or clavate, spore-bearing part 42–63 × 7–17 μm (x̄ = 52.86 × 11.76, n = 30), evanescent. Ascospores 9–12 × 8–10 μm (x̄ = 11.28 × 9.19, n = 90), biseriate, hyaline to olivaceous brown with age, limoniform, some globose, verruculose, thick-walled, biapiculate, bilaterally flattened, with an apical germ pore. Asexual morph: Undetermined.

Colony on PDA attaining 60 mm after 15 days at room temperature (25 ± 2 °C), circular, folded, white to light yellow, with slightly entire edge, sporulated within 30 days. Reverse yellow to yellow brown. Odor absent.

CHINA • Yunnan Province, Qujing City, Malong District, Wangjiazhuang Street, Y008, Longyangxian Cave, associated with decaying twigs on the floor of the cave, 25.434721°N, 103.509524°E, elev. ca 1920.43 m, 29 April 2024, Xiangfu Liu and Xuemei Chen, QJ2C2 (GMB-W1252), living culture GMBCC2271.

China (Wang et al. 2016b; Farr and Rossman 2026), Netherlands (Wang et al. 2016b), and Russia (Wang et al. 2016b, 2022).

Castanea mollissima (Farr and Rossman 2026), Glycine max (Farr and Rossman 2026), Homo sapiens (Wang et al. 2016b), Morus alba (Farr and Rossman 2026), unknown plant stem (Wang et al. 2016b), seed and dead stem of cereal (Wang et al. 2022).

Chaetomium subaffine was first described by Sergeeva (1961), but, lacking a Latin description, it was considered invalid. Then, Wang et al. (2022) reintroduced the species. In the phylogenetic analyses of the combined ITS, LSU, rpb2, and tub2 data, our isolates clustered with Chaetomium subaffine (CBS 637.91 (ex-type), and 22N617) within the Chaetomium clade, with 100% ML and 1.00 PP support (Fig. 20). Nucleotide comparisons between our strain (GMBCC2271) and C. subaffine (CBS 637.91, ex-type) showed 1.73% (10/578 bp, 1 gap) differences in ITS, 1.65% (10/607 bp, 1 gap) differences in LSU, 2.19% (11/503 bp, without gap) differences in rpb2, and 0.99% (7/704 bp, without gap) differences in tub2. Morphologically, the new isolate (GMBCC2271) resembles C. subaffine, with obovate or ovate ascomata usually covered by thick aerial hyphae, brown, erect to flexuous or slightly undulate terminal hairs, and brown, limoniform, biapiculate, bilaterally flattened ascospores (Wang et al. 2016b). Therefore, based on both morphology and phylogeny, we report Chaetomium subaffine as a new substrate record from a cave in Yunnan, China.

Trichocladium was introduced by Harz (1871), with Trichocladium asperum as the type species. Species of Trichocladium have been isolated from a wide range of substrates, including soil, estuarine sediments, leaves, dung, grass, meal, and Usnea cf. aurantio-atra (Wang et al. 2019). According to Index Fungorum (http://www.indexfungorum.org; accessed date 14 July 2026), the genus currently comprises 65 epithets, of which 43 are accepted species. Morphologically, Trichocladium is characterized by effuse colonies with mycelium partly superficial and partly immersed, absence of stromata, micronematous to semi-, macronematous, mononematous conidiophores that are straight or flexuous and pale brown, and solitary, dry, acrogenous or acropleurogenous, clavate to cylindrical, obovoid or pyriform, smooth to verrucose, thick-walled simple conidia, rounded at the apex, with one-to-many dark brown transverse septa (Park et al. 2017; Tagele et al. 2019). To date, more than six species of Trichocladium have been isolated from cave environments (Suppl. material 1: table S2). In the present study, Trichocladium cavernicola is described as a new species, representing the seventh cave-associated species known in this genus.

“Caverna” means cave in Latin, and “cola” means dweller or inhabitant, hence the creation of the epithet.

GMB-W1542.

Associated with bat guano in the cave environment. Asexual morph on PDA: Somatic hyphae usually hyaline, thin-walled, 2–4 μm wide, some becoming olivaceous brown and thick-walled when old, with conidia produced on the minimally differentiated conidiophores, on the top of hyphae or the side of vegetative hyphae. Conidiophores hyaline to subhyaline, unbranched, cylindrical, or slightly swollen on the top, 6–57 μm long, 2–8 μm wide near the top, sometimes lacking where the spores arise from an undifferentiated intercalary conidiogenous cell. Conidiogenous cell 6–23 × 3–8 μm (x̄ = 14.51 × 5.71 μm, n = 30), pale brown, terminal on conidiophores or intercalary in hyphae. Conidia 8–23 × 5–16 μm (x̄ = 13.57 × 11.15 μm, n = 120), single-celled, usually solitary, occasionally 2–3 in chains, olivaceous brown to dark olivaceous brown, smooth or slightly verruculose, globose or subglobose, sometimes ellipsoidal, occasionally pyriform, with a conspicuous apical, subapical or lateral germ pore. Sexual morph: Undetermined.

Colony on PDA attaining 60 mm after 15 days at room temperature (25 ± 2 °C), floccose, mouse gray to iron gray; reverse pale olivaceous gray to black. Colonies sporulated on PDA after 40 days. Odor absent.

CHINA • Yunnan Province, Kunming City, Fumin County, Chijiu Township, Tianshengqiao Village, Swallow Cave, bat guano, 25.388333°N, 102.537778°E, elev. ca 1864.03 m, 7 May 2021, Xiangfu Liu, FM32E (GMB-W1542, holotype), ex-type GZCC 25-0715; ibid., FM32E2 (GMB-W1543, isotype), ex-isotype GZCC 25-0716.

In phylogenetic analyses based on ITS, rpb2, and tub2 sequences, Trichocladium cavernicola (GZCC 25-0715 (ex-type), and GZCC 25-0716) formed a distinct lineage with 74% ML and 1.00 PP support, clustering as a sister taxon to Tr. solani, Tr. crispatum, and Tr. antarcticum (Fig. 20). Nucleotide comparisons between Tr. cavernicola (GZCC 25-0715, ex-type) and Tr. solani (VKM F-4902, ex-type) revealed 1.75% (10/572 bp, with 3 gaps) base pair differences in ITS, 2.37% (21/886 bp, with 2 gaps) differences in rpb2, and 2.08% (7/336 bp, with 2 gaps) differences in tub2. Morphologically, Tr. cavernicola is readily distinguished from Tr. solani by the latter having acremonium-like conidiophores and conidia formed basipetally in chains or curl up, that later aggregate into heads, and thick-walled, subglobose to angular chlamydospores composed of hyaline cells (Belosokhov et al. 2022). Trichoderma solani is a pathogenic fungus that causes potato tuber yellow rot (Belosokhov et al. 2022). In addition, the PHI test results (Fig. 6F) revealed no significant recombination relationships between Tr. cavernicola and its phylogenetically related taxa within the dataset. Based on both phylogenetic analyses and morphological characteristics, Trichocladium cavernicola is introduced as a new species.

Hypoxylaceae was introduced by de Lamarck and de Candolle (1805), but the name was not adopted in modern classifications and was long treated as a synonym of Xylariaceae (Hyde et al. 2020). Integrative studies incorporating multi-gene phylogeny, morphology, and chemotaxonomy have since validated Hypoxylaceae as a distinct family within Xylariales (Wendt et al. 2018; Daranagama et al. 2018). Species of Hypoxylaceae are predominantly saprobic or endophytic (Hyde et al. 2020). The sexual morph of Hypoxylaceae is characterized by globose to subglobose or elongate, cylindrical to pyriform ascomata embedded in stromata, arranged monostichously; the interior may be zonate or filled with liquid. Asci are unitunicate, cylindrical to clavate, and 4–8-spored. Ascospores are ellipsoidal to subglobose, brown to black, rarely hyaline, and mostly possess a germ slit (Hyde et al. 2020). The asexual morph is characterized by hyaline to light-brown, smooth, branched conidiophores, cylindrical conidiogenous cells, and hyaline, smooth to roughened, ellipsoidal conidia (Daranagama et al. 2018; Wendt et al. 2018). To date, ten species from six genera in this family have been documented from cave environments worldwide (Suppl. material 1: table S2).

Daldinia was introduced by Cesati and De Notaris (1863) and typified by Daldinia concentrica Ces. & De Not, named in tribute to the Swiss monk, Agostino Daldini (Cesati and De Notaris 1863; Yin et al. 2024). Daldinia is recognized as one of the largest genera in Hypoxylaceae (Xylariales, Sordariomycetes). The latest outline by Hyde et al. (2024) lists 56 accepted species in this genus, while 113 epithets are currently recorded in Index Fungorum (http://www.indexfungorum.org; 14 July 2026). Species of Daldinia typically occur on angiosperm hosts and are widely distributed in temperate and tropical regions, with more than 80% of known species occurring in tropical dry forests. In contrast, species found in temperate regions show a more restricted distribution and exhibit greater host specificity (Barbosa-Reséndiz et al. 2020; Liu et al. 2025). This genus is characterized by perithecia immersed in globose to claviform stromata, which are copper, violet, wine-colored, brown, or black, and release pigments that are ochraceous, purple, greenish, or gray when treated with KOH. Asci are cylindrical, stipitate, with an apical amyloid pore. Ascospores are ellipsoid, brown, and smooth to ornamented. The asexual morph is characterized by stout conidiophores and nodulisporium-like or periconiella-like anamorphs (Ju and Rogers 1996; Daranagama et al. 2015, 2018; Liu et al. 2025). In previous studies of cave-associated fungi, only one Daldinia species, D. concentrica, has been reported from caves in Slovenia and the UK. In this study, Daldinia eschscholtzii is reported as a new cave record.

Associated with rock surface in the cave environment. Asexual morph: Vegetative hyphae septate, branched, hyaline to brown, smooth-walled, 0.8–6.1 μm wide. Conidiophores sparse on aerial mycelium, straight or flexuous, erect or prostrate, smooth- and thin-walled, branched to unbranched, or reduced to conidiogenous cells borne laterally on hyphae. Conidiogenous cells mono- to polyblastic, blastic-sympodial, cylindrical, smooth- and thin-walled, 8.6–54.7 × 1.2–6.1 μm (x̄ = 27.53 × 3.22 μm, n = 30), with minute denticles after secession of conidia. Conidia 3.2–12.5 × 2.3–5.3 μm (x̄ = 6.82 × 3.30 μm, n = 60), cylindrical, reniform to lunate, or contracted at one end, hyaline to brown, aseptate, granulate, walls smooth and thin. Sexual morph: Undetermined.

Colonies on PDA attaining 60 mm after 7 days at room temperature (25 ± 2 °C), with undulate edge, white to pale yellow, sparse aerial mycelium on the surface; reverse pale yellow. The white reproductive mycelium covers the stromatic colonies after 50 days.

CHINA • Yunnan Province, Nujiang Lisu Autonomous Prefecture, Lushui City, Daxingdi Town, rock surface in Walaya Cave, 26.080743°N, 98.848391°E, elev. ca 850.46 m, 23 April 2024, Xiangfu Liu and Xuemei Chen, NJ9A (GMB-W1474), living culture, GMBCC2475.

Australia, Bahamas, Bangladesh, Brazil, Brunei Darussalam, Cook Islands, Cuba, Democratic Republic of the Congo, Federal States of Micronesia, Ghana, Japan, Kenya, Malaysia, New Zealand, Nigeria, Pakistan, Papua New Guinea, Philippines, Saint Lucia, Seychelles, Sierra Leone, Singapore, Tanzania (Farr and Rossman 2026), China (Farr and Rossman 2026; this study), France, Germany, Mexico, South Africa (Jayasiri et al. 2015), India, Indonesia, Samoa, Trinidad and Tobago, the USA (Jayasiri et al. 2015; Farr and Rossman 2026), and Thailand (Jayasiri et al. 2015; Brooks et al. 2024; Farr and Rossman 2026).

Cordia sebestena, Acacia sp., Albizia lebbeck, Albizia sp., Albizia zygia, Angelica sp., Annona muricata, Antidesma sp., Artocarpus altilis, Artocarpus integrifolius, Brguiera sexangula, Bucida palustris, Bursera simaruba, Cassia floribunda, Casuarina sp., Celtis kraussiana, Citrus aurantiifolia, Citrus aurantium, Citrus paradisi, Citrus sp., Citrus limon, Citrus nobilis, Cocos nucifera, Cordia sebastina, Croton sp., Dendrobium officinale, Entandrophragma cylindricum, Eucalyptus alba, Eucalyptus sp., Excoecaria agallocha, Ficus sp., Gliricidia sepium, Heteromorpha trifoliata, Hevea brasiliensis, Hevea sp., Hippomane mancinella, Jambosa vulgaris, Mangifera indica, Morus alba, Musa sp., Neohypodiscus cerebrinus, Nothofagus sp., Pinus sp., Platanus wrightii, Pogostemon cablin, Quercus acutissima, Quercus sp., Sizygium jambos, Tamarindus indica, wood, and Xestospongia sponge species (Farr and Rossman 2026), Oncosperma sp. (Brooks et al. 2024), and rock surface in cave habitat (this study).

Daldinia eschscholtzii was first reported by Rehm (1904) based on its sexual morphological characters, including elliptic to falcate ascospores, measuring 10–12 × 5 μm, from specimens collected in the USA. Our new isolate (GMBCC2475) shows morphological similarities to D. eschscholtzii (MFLUCC 20–0232), sharing obovoid to ellipsoid, aseptate, smooth conidia, often with a truncate base, and conidiophores forming a dichotomous to trichotomous conidiogenous apparatus with a Nodulisporium-like branching pattern, in which 1–3 conidiogenous cells arise from each terminal branch (Samarakoon et al. 2019; Gajanayake et al. 2021). Multi-gene phylogeny of the combined ITS, LSU, tub2 and rpb2 sequences further showed that our isolate clusters with D. eschscholtzii and is clearly separated from other species with 100% ML and 1.00 BYPP support (Fig. 24). Therefore, based on both morphology and phylogeny evidence, we report this isolate as a new substrate record of D. eschscholtzii from a cave in Yunnan, China.