Taxonomy, phylogeny and diversity of hyperparasites and their allied entomopathogens in Hypocreales

DP Wei; E Gentekaki; JJ Luangsa-ard; NN Wijayawardene; DN Wanasinghe; ZL Liu; QF Huang; NY Liu; SW Xie; JC Kang; TC Wen

doi:10.3114/sim.2026.114.02

. 2026 May 20;114:370–436. doi: 10.3114/sim.2026.114.02

Taxonomy, phylogeny and diversity of hyperparasites and their allied entomopathogens in Hypocreales

DP Wei ^1,², E Gentekaki ³, JJ Luangsa-ard ⁴, NN Wijayawardene ⁵, DN Wanasinghe ^6,⁷, ZL Liu ^2,⁸, QF Huang ², NY Liu ², SW Xie ^2,⁸, JC Kang ^1,², TC Wen ^1,^2,^*

PMCID: PMC13487413 PMID: 42621338

Abstract

Hyperparasitic fungi associated with hypocrealean entomopathogens play a pivotal yet underexplored role in regulating fungal-arthropod interaction networks. Despite their frequent occurrence in nature, the taxonomic diversity and evolutionary relationships of these hyperparasites remain poorly resolved. In this study, we conducted an integrative taxonomic investigation of hyperparasites associated with hypocrealean entomopathogenic fungi, together with their closely allied entomopathogenic taxa. Multilocus phylogenetic analyses based on LSU, ITS, SSU, tef-1α, rpb1, and rpb2 were employed to reconstruct phylogenetic relationships, and molecular clock analyses were conducted independently to infer evolutionary timescales of families within Hypocreales. Morphological examinations were used to corroborate species delimitation and generic circumscription. Our results substantially expand the known diversity of this group, revealing one novel family, Chlorocilliaceae fam. nov., two new genera (Paralecanicillium gen. nov. and Neodingleyomyces gen. nov.), and 18 new species. In addition, three new combinations, seven new host, geographic, or asexual morph records, and two previously described species are documented, comprising a total of 30 species distributed across 18 genera and six families. Chlorocilliaceae is proposed to accommodate Albacillium, Chlorocillium, Fiorinimazzantia, Husseyia, Neoaraneomyces, Paraneoaraneomyces, Rousseaua, Speluncomyces and Subuliphorum. Updated descriptions and emended generic concepts of Fiorinimazzantia and Husseyia are provided based on newly collected material. This study advances our understanding of the taxonomy, evolutionary history, and hidden diversity of hyperparasitic fungi allied to hypocrealean entomopathogens, and provides a robust framework for elucidating tripartite host-pathogen-hyperparasite interactions in fungal ecology.

Taxonomic novelties: New family: Chlorocilliaceae D.P. Wei & T.C. Wen. New genera: Neodingleyomyces D.P. Wei, Z.L. Liu & T.C. Wen, Paralecanicillium D.P. Wei & T.C. Wen. New species: Ascopolyporus sanduensis D.P. Wei & T.C. Wen, Calcarisporium ophiocordycipiticola D.P. Wei & T.C. Wen, Chlorocillium aschersoniae D.P. Wei & T.C. Wen, Chlorocillium coccidiicola D.P. Wei & T.C. Wen, Chlorocillium cordycipiticola D.P. Wei, Z.L. Liu & T.C. Wen, Chlorocillium gibellulae D.P. Wei & T.C. Wen, Chlorocillium globiceps D.P. Wei & T.C. Wen, Chlorocillium houqiaoense D.P. Wei, Q. F. Huang & T.C. Wen, Chlorocillium menghaiense D.P. Wei & T.C. Wen, Chlorocillium puerense D.P. Wei, Z.L. Liu & T.C. Wen, Chlorocillium sanduense D.P. Wei & T.C. Wen, Chlorocillium taiyangheense D.P. Wei & T.C. Wen, Chlorocillium tengchongense D.P. Wei, N.Y. Liu & T.C. Wen, Husseyia ramosissima D.P. Wei & T.C. Wen, Neodingleyomyces tengchongensis D.P. Wei, N.Y. Liu & T.C. Wen, Ophiocordyceps shimenxiaensis D.P. Wei & T.C. Wen, Paralecanicillium yunnanense D.P. Wei & T.C. Wen, Pleurocordyceps simaoensis D.P. Wei & T.C. Wen. New combinations: Calcarisporium andense (Flakus et al.) D.P. Wei & T.C. Wen, Calcarisporium agaricicola (Berk.) D.P. Wei & T.C. Wen, Torrubiellomyces sessilis (Kaitsu et al.) D.P. Wei, T.C. Wen. New records: Albacillium hingganense M.M. Ding & L.J. Xu, Chlorocillium neolepidopterorum (W.H. Chen et al.) W.H. Chen et al., Fiorinimazzantia elisabettae Y.P. Tan et al., Pleurocordyceps heilongtanensis Y.B. Wang et al., Pleurocordyceps parvicapitata Y.P. Xiao et al., Pleurocordyceps neoagarica Yu Yang & Yuan Pin Xiao, Paradingleyomyces lepidopterorum Y. Wang tris & T.C. Wen. Replacement names: Torrubiellomyces J.P.M. Araújo & de Bekker.

Citation: Wei DP, Gentekaki E, Luangsa-ard JJ, Wijayawardene NN, Wanasinghe DN, Liu ZL, Huang QF, Liu NY, Xie SW, Kang JC, Wen TC (2026). Taxonomy, phylogeny and diversity of hyperparasites and their allied entomopathogens in Hypocreales. Studies in Mycology 114: 370–436. doi: 10.3114/sim.2026.114.02

Keywords: Entomopathogenic fungi, hyperparasitism, integrative taxonomy, new taxa

Introduction

Hyperparasitic fungi occupy distinct ecological niches, playing a crucial role in regulating host population dynamics and transmission rates within natural ecosystems (Bermúdez-Cova et al. 2023). Conversely, in cultivated environments, certain hyperparasitic species such as Calcarisporium cordycipiticola and Lecanicillium coprophilum cause substantial economic losses to the mushroom industry (Sun et al. 2016, Liu et al. 2021a, Nguyen et al. 2023). Despite their evident ecological impact and economic significance, hyperparasitic fungi remain largely understudied, particularly those targeting entomopathogenic fungi (Araújo et al. 2022, Bermúdez-Cova et al. 2023).

Hypocreales (Sordariomycetes, Ascomycota) comprises 28 families (Samarakoon et al. 2022, Hyde et al. 2024), with entomopathogenic species predominantly concentrated within the Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae, and Polycephalomycetaceae (Sung et al. 2007, Chaverri et al. 2008, Kryukov et al. 2017, Wen et al. 2017, Xiao et al. 2018, 2019, 2023, Mongkolsamrit et al. 2020, Wang et al. 2020, Wei et al. 2019, 2020, 2021, Peng et al. 2024). Species in these families can infect various developmental stages of insects including eggs, larvae, pupae, and adults (Wei et al. 2022). After penetrating the insect cuticle, hyphae of entomopathogenic fungi proliferate in the hemocoel, consuming the insect’s nutrients, eventually leading to the insect’s death and the fungus producing spores on the cadaver (Ballesteros-Aguirre et al. 2025). Hypocrealean entomopathogenic fungi commonly reproduce by means of forming sexual and/or asexual structures (Ballesteros-Aguirre et al. 2025). Generally, the sexual structures of entomopathogenic fungi possess stalked or sessile stroma bearing a terminal, lateral or intercalary fertile part (Wei et al. 2022). In contrast, the asexual structure is made by anastomosing mycelial mats on the cadaver, from which synnemata may or may not arise (Chen et al. 2025a). However, despite being sophisticated parasites themselves, some entomopathogenic species are also parasitized by other hyperparasitic fungi. Hyperparasitism is a common antagonistic interaction involving direct contact, wherein hyperparasites coil around, penetrate, and lyse host hyphae to absorb nutrients (Hasan et al. 2022). This complex physiological process relies on the production of hydrolytic enzymes (e.g., cellulases, chitinases, proteases) and secondary metabolites to degrade host cell walls (Mukherjee et al. 2022). Hyperparasites adopt either a biotrophic strategy, utilizing haustoria to extract nutrients from living tissue, or a necrotrophic strategy, secreting antifungal compounds to kill the host and destroy cellular integrity (Bermúdez-Cova et al. 2023). Eventually, the spore-producing organs of infected entomopathogenic fungi will be sabotaged and fail to sporulation, in which hyperparasitic fungi play a vital role in maintaining the balance between entomopathogenic fungi and their insect hosts (Andersen et al. 2012, Bermúdez-Cova et al. 2023).

Previous studies indicate that entomopathogen-associated hyperparasitic fungi are taxonomically concentrated and ecologically specialized. To date, at least 17 species have been documented on the stromata of Ophiocordyceps and Paraisaria, with the majority belonging to Polycephalomycetaceae, particularly the genera Polycephalomyces, Pleurocordyceps, and Perennicordyceps (Ban et al. 2009, Wang et al. 2014, Xiao et al. 2018, 2023, 2024, Araújo et al. 2022, Wei et al. 2022). Beyond Polycephalomycetaceae, additional hyperparasitic lineages have been reported sporadically from Cordycipitaceae and related families, including species of Samsoniella, Simplicillium, Niveomyces, and Pseudoniveomyces, parasitizing stromata of diverse Ophiocordyceps hosts (Wei et al. 2019, Wang et al. 2020, Araújo et al. 2022, Kobmoo et al. 2023). Collectively, these studies suggest that hyperparasitism on entomopathogenic fungi has evolved multiple times across Hypocreales but remains unevenly documented, with most taxa known only from limited host associations. Notably, the holomorph of these fungi are still poorly understood, as the vast majority of described hyperparasites are known exclusively from their asexual morphs, with a confirmed sexual morph reported only for Torrubiellomyces zombiae on Ophiocordyceps camponoti-floridani (Araújo et al. 2022).

It is noteworthy that the stromata or colonies of entomopathogenic fungi constitute a unique ecological niche harbouring diverse fungal taxon. Most previously described hyperparasitic fungi belong to Polycephalomycetaceae and have predominantly been documented on Ophiocordyceps stromata, whereas reports from other hypocrealean entomopathogenic genera remain scarce, suggesting that substantial hidden diversity may still exist on this specialized substrate. To address this knowledge gap and resolve existing taxonomic uncertainties, this study undertook extensive diversity surveys and sampling of hyperparasitic fungi associated with hypocrealean entomopathogens in China, which provided the foundation for subsequent phylogenetic analyses. These analyses were used to clarify evolutionary relationships, identify hyperparasites and their closely related entomopathogenic fungi, and reveal phylogenetically unresolved lineages. Based on integrative evidence from morphology and phylogeny, we further refined generic concepts, clarified the taxonomic placement of problematic genera such as Fiorinimazzantia, Husseyia, and Rousseaua, and corrected previous taxonomic misclassifications. Collectively, this work aims to establish a coherent taxonomic and evolutionary framework for hyperparasitic fungi associated with entomopathogenic hosts, thereby contributing to the resolution of long-standing taxonomic controversies and improving our understanding of their ecological and evolutionary relationships.

Materials and methods

Sampling and isolation

Field collections were conducted in mixed evergreen and deciduous broad-leaved forests in Guizhou, Jilin, and Yunnan provinces, China, representing southern, northeastern, and southwestern regions of the country (Fig. 1). Sampling was carried out between 2018 and 2024 and covered multiple seasons (June–November) to capture potential temporal variation in the occurrence of hyperparasitic fungi. Specimens were collected from the undersides of living leaves, leaf litter, and forest floors. A total of 63 specimens were collected, representing 11 genera in seven families of Hypocreales, including 37 hyperparasitic fungal specimens and 26 entomopathogenic fungal specimens. Detailed geographic and environmental data, including GPS coordinates, elevation, ambient temperature, habitat type, and collection date, were recorded in situ. All specimens were photographed in the field using an iPhone XS Max equipped with a camera application (http://www.dddsway.com/) to document macromorphological features and environmental context.

Fig. 1 — Localities of collecting sites and genus composition of studied fungi in China. Green shading indicates sampled provinces (Jilin, Guizhou, and Yunnan). Coloured pie charts represent the relative abundance of fungal genera at each site; numbers in parentheses indicate total specimens per genus.

Fresh specimens were individually placed in sterile plastic boxes, transported to the laboratory, and processed within 24 h. Prior to isolation, hyperparasitic fungal (HPF) structures were examined by observing the surfaces of stromata or synnemata of entomopathogenic fungi (EMP) to determine the presence and distribution of infection. Hyperparasitic fungi were isolated using single-spore or tissue isolation methods, depending on the developmental stage of the fungus and the availability of diagnostic structures (Fig. 2). For Pleurocordyceps species producing abundant, tightly adhering conidia, single-spore isolation was employed. Conidial masses were carefully removed from HPF-infected stromata or synnemata using a sterile needle, suspended in 100 μL of sterile distilled water, gently agitated, and spread onto potato dextrose agar (PDA) plates. Plates were incubated at 25 °C in dark for 24–48 h, after which germinating single spores were transferred to fresh PDA plates using a stereomicroscope to obtain pure cultures. For taxa with powdery or sparse conidia and for Ophiocordyceps species with mycelia developing inside the insect host, tissue isolation was applied. Small fragments (approximately 1–2 mm²) of infected stromata or synnemata were excised using sterile tweezers, placed onto PDA plates, and incubated at 25 °C in dark for 3–10 d. During this period, growth was observed daily. Contaminated cultures were purified and incubated for an additional 15–30 d.. To isolate insect-associated Ophiocordyceps, host bodies were surface-sterilized with cotton swabs soaked in 75 % ethanol for 30 s. After ethanol evaporation, the cuticle was aseptically removed, and internal mycelia were extracted and transferred to fresh PDA plates, followed by incubation at 25 °C in dark for 30–60 d. Emerging colonies were repeatedly subcultured onto fresh PDA plates at 25 °C in dark for 15–30 d until pure cultures were obtained. Colony morphology was photographed after 30 d of incubation. Pure cultures were preserved by transferring mycelium and agar plugs into sterile distilled water-filled centrifuge tubes and deposited in the Kunming Institute of Botany Culture Collection (KUNCC). Voucher specimens were dried using allochroic silica gel and deposited in the Herbarium of Cryptogams, Kunming Institute of Botany (KUN), Chinese Academy of Sciences, Kunming, China. Newly introduced taxa were registered in MycoBank (Crous et al. 2004).

Fig. 2 — Schematic overview of the isolation, purification, and preservation procedures used for entomopathogenic fungi (EMP) and associated hyperparasitic fungi (HPF) The abbreviation "SW" refers to sterilized water."

Morphological observation

Colonies and synnemata on their natural substrate were examined and photographed using a Leica S9E dissecting microscope equipped with a digital camera. Mycelia carrying sporulation structures were taken from dried specimens or living cultures and mounted on microscope slides using water, cotton blue solution, or Congo red solution. The micro-structures, such as perithecia, asci, ascospores, conidiophores, phialides and conidia, were observed using a Nikon Eclipse 80i compound microscope, and the microphotographs were taken with a DS-Ri2 camera fitted on the compound microscope. Cultural characteristics were recorded using an iPhone XS Max camera, with colour codes referenced from the Methuen Handbook of Colour (Kornerup & Wanscher, 1978). Images were processed using Adobe Photoshop CS3 Extended v. 10.0 (Adobe, San Jose, CA). The Tarosoft (R) Image Frame Work program was used to take measurements.

DNA extraction, PCR amplification and sequencing

Genomic DNA was extracted from fresh mycelia grown on PDA, or mycelia/fruiting bodies on natural substrate using E.Z.N.A.TM Fungal DNA MiniKit (Omega Biotech, CA, USA) following the manufacturer’s instructions. Six genetic markers including the internal transcribed spacer (ITS1-5.8S-ITS2) (ITS) of the nuclear rDNA, the nuclear large subunit rDNA (LSU), the nuclear small subunit rDNA (SSU), the translation elongation factor 1-α (tef-1α), the RNA polymerase II largest subunit (rpb1) and RNA polymerase II second largest subunit (rpb2) were amplified using primers described in Wei et al. (2021). The PCR amplifications of ITS, LSU and SSU were performed in 25 μL reaction volumes consisting of 12.5 µL PCR mixture (2 × Taq PCR Master Mix, red dye), 8.5 µL double-sterilized water, 1 µL (at 10 uM/L final concentration) of each primer, and 3 µL DNA template. BioRad C1000 thermal cycler (Bio-Rad Laboratories Inc., Hercules, CA, USA) was used to amplify the ITS, LSU and SSU as follows: Initialization at 95 °C for 5 min, followed by denaturation at 95 °C for 30 s, annealing at 56 °C for 50 s, and extension at 72 °C for 50 s, followed by a final extension at 72 °C for 10 min. The PCR amplifications of tef-1α, rpb1, and rpb2 were performed in a 50 μL volume reaction consisting of 25 µL PCR mixture (2 × Taq PCR Master Mix, red dye), 17 µL double-sterilized water, 2 µL of each primer, and 4 µL DNA template. Thermocycler conditions of tef-1α, rpb1, and rpb2 were as follows: Initialization at 95 °C for 5 min, 10 cycles of denaturation at 95 °C for 30 s; annealing at 60 °C for 1 min for tef-1α and rpb1 and at 56 °C for 1 min for rpb2 and extension at 72 °C for 1 min, followed by 35 cycles of denaturation at 95 °C for 30 s, annealing at 52 °C for 1 min, and extension at 72 °C for 1 min with a ﬁnal elongation at 72 °C for 10 min. Amplicons were checked with 1 % ethidium bromide-stained agarose gel under UV light and sent to Sangon Biotech company (Shanghai, China) for sequencing. The newly generated sequences were submitted to GenBank for assignment of accession numbers.

Phylogenetic analyses

Sequence data generated in this study were assembled using Sequencing Project Management (SeqMan) (Clewley 1995) or BioEdit v. 7.1.8 (Hall 1999). The assembled sequences were used as queries in BLAST searches against GenBank to confirm the absence of opportunistic fungal contamination and to guide taxon sampling. Preliminary BLAST results indicated that the newly generated hyperparasitic fungi and their hosts were distributed across five families of Hypocreales, namely Cordycipitaceae, Clavicipitaceae, Calcarisporiaceae, Ophiocordycipitaceae, and Polycephalomycetaceae. Based on these results, four phylogenetic datasets were constructed: (1) a familial-level dataset of Hypocreales comprising 369 taxa representing 28 families (Table S1), with denser sampling of Ascopolyporus, Calcarisporium, Clavicipitaceae, Cordycipitaceae, and Polycephalomycetaceae to resolve the placement of newly introduced taxa; (2) a Chlorocillium dataset including 75 taxa (Table S2); (3) a Pleurocordyceps dataset including 64 taxa (Table S3); and (4) an Ophiocordyceps dataset including 149 taxa (Table S4). The latter three datasets were more densely sampled to achieve finer resolution of interspecific relationships. Each dataset comprised six loci, namely LSU, ITS, SSU, tef-1α, rpb1, and rpb2. Individual loci were aligned independently using MAFFT v. 7 (Kuraku et al. 2013, Katoh et al. 2019), and ambiguously aligned regions were automatically trimmed using TrimAl v. 1.2 on the Phylemon web server (Capella-Gutiérrez et al. 2009) with a gap threshold (-gt) of 0.6. The six loci were concatenated using SequenceMatrix v. 1.7.8 (Vaidya et al. 2011), and the resulting alignments were used for phylogenetic analyses. Maximum likelihood (ML) analyses were performed using IQ-TREE v. 2.2.0 (Nguyen et al. 2015) implemented in PhyloSuite v. 1.2.3 (Zhang et al. 2020, Xiang et al. 2023). An edge-linked partition model was applied, and branch support was assessed using 5000 ultrafast bootstrap replicates (UFB) (Minh et al. 2013) and the Shimodaira-Hasegawa-like approximate likelihood-ratio test (SH-aLRT) (Guindon et al. 2010). The best-fit substitution models for each partition was selected using ModelFinder under the Bayesian information criterion (BIC) (Chernomor et al. 2016, Kalyaanamoorthy et al. 2017). In ML analyses, node support was considered strong when both SH-aLRT ≥ 80 and UFB ≥ 95, moderate when either SH-aLRT < 80 or UFB < 95, and weak when both values were below these thresholds (Wei et al. 2022). Bayesian inference (BI) was conducted using MrBayes v. 3.2.7a (Ronquist et al. 2012). The best-fit nucleotide substitution models were selected using MrModeltest v. 2.2 (Nylander 2004). For the familial-level dataset, six Markov chains were run for 30 M generations with sampling every 5000 generations and a burn-in of 25 %. For the remaining three datasets, six Markov chains were run until the average standard deviation of split frequencies fell below 0.01, with sampling every 1000 generations and a burn-in of 25 %. Bayesian posterior probabilities (PP) were calculated from the remaining 75 % of sampled trees, and nodes with PP ≥ 0.90 were considered strongly supported. Resulting trees were visualized using FigTree v. 1.4.4 (Rambaut 2018) and edited in Adobe Illustrator v. 2020.

Genealogical concordance phylogenetic species recognition (GCPSR)

A pairwise homoplasy index (PHI) test was performed in SplitsTree6 v. 6 to assess potential recombination among closely related strains within Chlorocillium (Huson & Bryant 2024). The input datasets comprised 26 taxa for clade C1 (species of the Chlorocillium guizhouense complex), ten taxa for clade 2 (Ch. neolepidopterorum and Ch. menghaiense) six taxa for each of clade C3 (Ch. mauryae, Ch. gibellulae, Ch. houqiaoense), and clade C4 (Ch. neorongjiangense, Ch. tengchongense, Ch. cordycipiticola). Analyses were based on concatenated sequences of six loci (LSU, ITS, SSU, tef-1α, rpb1, and rpb2), with a total alignment length of 5091 base pairs. Sequence alignments were converted from FASTA to NEXUS format using the ALTER web server (https://www.sing-group.org/ALTER/) and subsequently imported into SplitsTree6. Split networks were constructed from the concatenated datasets using the LogDet transformation (Steel 1994) and the split decomposition method (Bandelt & Dress 1992). Statistical significance of recombination was evaluated using the Φw statistic, with values below 0.05 (Φw < 0.05) interpreted as evidence of significant recombination, indicating a lack of genealogical exclusivity among the tested lineages.

Divergence time estimation

Molecular dating has increasingly been applied as a complementary approach for evaluating higher-rank classification in fungi when interpreted together with phylogenetic and morphological evidence (Sung et al. 2008, Hongsanan et al. 2017, Bao et al. 2023). Within Sordariomycetes, lineages possessing stem ages of approximately 50–150 M yr ago (MYA) are generally considered to correspond to family-level taxa (Hyde et al. 2017). In the present study, we have assembled a phylogenetic dataset containing 207 taxonomic units representing the 28 families of Hypocreales (Hypocreomycetidae) and taxa of three subclasses of Sordariomycetes, such as Diaportheomycetidae, Sordariomycetidae, Xylariomycetidae. Taxa used for divergence time estimation (Table S5) were prepared by excluding redundant taxa from the 369-taxa Hypocreales dataset, with dense sampling within Ophiocordyceps, following the approach of Zhuang et al. (2025). The fossil Paleoophiocordyceps gerontoformicae was used to calibrate the stem node of the myrmecophilous hirsutelloid clade, using a lognormal prior (offset = 99, mean = 1.5, SD = 1) (Zhuang et al. 2025). The crown age of Xylariomycetidae was applied as a secondary calibration point using a normal distribution (mean = 168 MYA, SD = 16), following Hongsanan et al. (2017), Samarakoon et al. (2022), and Yu et al. (2024). Divergence time estimation among families of Hypocreales was conducted using the BEAST v. 2.6.3 program. The aligned data matrix used for ML and BI analyses was partitioned using Aliview v. 1.28 (Larsson 2014) and loaded to BEAUti v. 2.2.x for constructing an XML ﬁle. The best-fit substitution models were selected by ModelFinder (Kalyaanamoorthy et al. 2017) with Bayesian information criterion (BIC) implemented in OFPT v. 1.9.0 (Zeng et al. 2023), resulting in the selection of TN+F+R4 for LSU, TIM3e+R4 for SSU, TIM2+F+R6 for tef-1α, GTR+F+I+G4 for rpb1 and GTR+F+I+G4 for rpb2. However, TN, IM3e and TIM2 are not implemented in BEAUti v. 2.2.x, the GTR model was used as a substitute for these partitions, with base frequencies set to “Empirical”, following the BEAST v. 2.3.6 tutorial. Site models, clock models, and tree models were linked across all partitions. A strict clock model was applied, and a Calibrated Yule model was used as the tree prior to model speciation. Analyses were run for 30000 M generation, with parameters sampled every 5000 generations. The effective sample sizes (ESS) were checked in Tracer v. 1.6 (Rambaut et al. 2013), wherein ESS values of all parameters greater than 200 were acceptable. The first 20 % of trees representing burn-in phase were discarded based on Tracer v. 1.6 and the remaining 80000 trees were combined with LogCombiner v. 1.8.0 to generate a maximum clade credibility (MCC) tree using TreeAnnotator v. 1.8.0. The molecular dating tree was displayed with Figtree v. 1.4.4 (Rambaut 2018) and edited with Adobe Illustrator CS6 (Adobe Systems Inc., USA).

Results

Phylogenetic analyses

Analyses 1 – The familial placements of Hypocreales

The Hypocreales 369-taxa dataset contained 4855 characters including gaps (LSU: 833 bp, ITS: 518 bp, SSU: 1020 bp, tef-1α: 902 bp, rpb1: 668 bp, rpb2: 914 bp) with 2244 parsimony-informative, 484 singleton, and 2127 constant sites. Allocryptovalsa sichuanensis, Fasciatispora cocoes, and Nemania thailandensis within Xylariales were selected as the outgroup taxa. All families were monophyletic with strong statistical support with exception of Clavicipitaceae (Fig. 3). In the maximum likelihood analysis (lnL = −170294.904), the newly sequenced taxa were placed within six families: Calcarisporiaceae, Chlorocilliaceae fam. nov., Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae, and Polycephalomycetaceae.

Fig. 3 — Maximum likelihood (IQ-TREE) phylogeny of *Hypocreales* based on a concatenated dataset (LSU, ITS, SSU, *tef-1α*, *rpb1*, and *rpb2*). Family names are denoted at the right side of the tree. The newly obtained strains are indicated with red star. The hyperparasitic fungi on entomopathogenic fungi are highlighted in purple text. SH-aLRT value (left) and the UFB value (right) equal to or greater than 50 and posterior probabilities (middle) equal to or higher than 0.90 are given near to the nodes. Superscript ‘T’ denotes the type strains/species. Hyperparasites are denoted with purple asterisks.

In Calcarisporiaceae, the new collections HKAS 148551 and HKAS 148552 form a monophyletic clade with Neobaryopsis andensis and Neobarya agaricicola (SH-aLRT = 88 %, PP = 0.96, UFB = 78 %) and cluster with other species of Calcarisporium. The newly proposed family Chlorocilliaceae, consisting of nine genera (i.e. Albacillium, Chlorocillium, Fiorinimazzantia, Husseyia, Neoaraneomyces, Paraneoaraneomyces, Rousseaua, Speluncomyces and Subuliphorum) and is sister to Clavicipitaceae (SH-aLRT = 98 %, PP = 1, UFB = 100 %). Within Chlorocilliaceae, the collection HKAS 135605 groups with Fiorinimazzantia elisabettae, which forms a distinct clade. The collection HKAS 135601 branches off from a clade of Husseyia annamariae and H. queenslandica with maximum support. Three collections (HKAS 135603, HKAS 148545, and HKAS 148544) cluster with Albacillium hingganense, and 44 specimens fall within Chlorocillium (their interspecific relationships illustrated in Fig. 4). In Cordycipitaceae, newly collected taxa, HKAS 148575 and HKAS 148576, form a distinct clade that is sister to Ascopolyporus galloides with robust support (SH-aLRT = 99 %, PP = 1.00, UFB = 100 %). Specimens HKAS 148556, HKAS 148558 and HKAS 148557 cluster into a distinct clade sister to species of Pleurodesmospora coccorum with significant support (SH-aLRT = 65 %, UFB = 93 %). In Clavicipitaceae, the specimen HKAS 148550 aligns with Metarhizium puerense with strong support (SH-aLRT = 94 %, PP = 1.00, UFB = 100 %), indicating a conspecific relationship. In Ophiocordycipitaceae, the host portion of HKAS 148571, along with two specimens (HKAS 148569 and HKAS 148572), forms a distinct clade with high support (SH-aLRT = 97 %, PP = 1, UFB = 100 %) within the Ophiocordyceps pseudoacicularis complex. In Polycephalomycetaceae, specimens HKAS 148565 and HKAS 148563 form a clade sister to Dingleyomyces with strong support (SH-aLRT = 98 %, PP = 1, UFB = 100 %). Specimen HKAS 148571 groups with Paradingleyomyces lepidopterorum, and seven specimens (HKAS 148573, HKAS 148554, HKAS 148549, HKAS 148555, HKAS 148553, HKAS 148546, HKAS 148550Y) are nested within Pleurocordyceps, with its interspecific relationship illustrated in Fig. 5.

Fig. 4 — Maximum likelihood (IQ-TREE) phylogeny of *Chlorocillium* based on the combined LSU, ITS, SSU, *tef-1α*, *rpb1*, and *rpb2* sequence dataset. The newly obtained strains are indicated in blue text. SH-aLRT value (left) and the UFB value (right) equal to or greater than 50 and posterior probabilities (middle) equal to or higher than 0.90 are given near to the nodes. The branches of the *Ch. guizhouense* complex are highlighted in red. Superscript ‘T’ denotes the type strains/specimens. Pairwise homoplasy index (PHI) test of closely related species using both LogDet transformation and splits decomposition are presented in the left side of the tree. PHI test results (Φw) < 0.05 indicate significant recombination among the species.

Fig. 5 — Maximum likelihood (IQ-TREE) phylogeny of *Pleurocordyceps* based on the combined LSU, ITS, SSU, *tef-1α*, *rpb1*, and *rpb2* sequence dataset. The newly obtained strains are indicated in blue text. SH-aLRT value (left) and the UFB value (right) equal to or greater than 50 and posterior probabilities (middle) equal to or higher than 0.90 are given near to the nodes. Superscript ‘T’ denotes the type strains/specimens.

Analyses 2 - Interspecific relationship of Chlorocillium

The dataset of 68 strains and/or specimens of Chlorocillium comprises 5091 characters including gaps (LSU: 840 bp; ITS: 544 bp; SSU: 1021 bp; tef-1α: 908 bp; rpb1: 744 bp; rpb2: 1034 bp) with 668 parsimony-informative, 276 singleton, and 4147 constant sites. Husseyia annamariae, H. queenslandica, and H. ramosissima were treated as the outgroup taxa. The best-scoring ML tree with a final likelihood value of −14870.796438 is presented in Fig. 4.

Thirteen spider-pathogenic fungal specimens cluster with Chlorocillium guizhouense (SH-aLRT = 90 %, UFB = 87 %, Fig. 4). Specimens HKAS 135599 and HKAS 148559 form a clade sister to Ch. guizhouense. Five specimens of Chlorocillium globiceps, exclusively hyperparasitic on Ophiocordyceps cf. globiceps, constitute a strongly supported clade (SH-aLRT = 99 %, PP = 1.0, UFB = 100 %, Fig. 4) sister to Chlorocillium taiyangheense (HKAS 148568 and HKAS 135602). Chlorocillium sanduense forms a well-defined clade (SH-aLRT = 100 %, PP = 1.0, UFB = 100 %, Fig. 4) that branches off from Ch. guizhouense complex. Nine specimens associated with diverse entomopathogenic fungal hosts cluster with Ch. neolepidopterorum with moderate support from maximum likelihood analysis (SH-aLRT = 99 %, UFB = 70 %, Fig. 4) that is distantly related to other congeners. Chlorocillium houqiaoense, Chlorocillium gibellulae, and Ch. mauryae form a well-supported monophyletic clade (SH-aLRT = 92 %, PP = 1, UFB = 99 %, Fig. 4) separate from other species of Chlorocillium. Chlorocillium puerense is resolved as sister to Chlorocillium montefioreae with strong support (SH-aLRT = 99 %, PP = 1.0, UFB = 100 %, Fig. 4). A maximally supported clade comprising Ch. tengchongense, Ch. neorongjiangense, and Ch. cordycipiticola was resolved as a lineage distantly related to other species of the genus. Chlorocillium coccidiicola was placed in a basal clade to other species in the genus with maximum support.

Analyses 3 - Interspecific relationship of Pleurocordyceps

The dataset of 65 strains and/or specimens of Pleurocordyceps comprises 5035 characters including gaps (LSU: 845 bp; ITS: 563 bp; SSU: 1017 bp; tef-1α: 892 bp; rpb1: 675 bp; rpb2: 1043 bp) with 854 parsimony-informative, 123 singleton, and 4058 constant sites. Five taxa of Perennicordyceps were selected as the outgroup. The best-scoring ML tree with a final likelihood value of −16340.716 is presented in Fig. 5.

Specimens HKAS 148553 and HKAS 148546 cluster as a sister clade to Pleurocordyceps sanduensis with strong support (SH-aLRT = 88 %, PP=0.99, UFB = 97 %, Fig. 5). Specimens HKAS 148573, HKAS 148554, HKAS 148574 and HKAS 148549 together with Pleurocordyceps heilongtanensis are placed in a distinct clade (SH-aLRT = 99 %, PP = 0.99, UFB = 100 %, Fig. 5). Specimens HKAS 148555 and HKAS 148550Y are resolved as Pleurocordyceps parvicapitata and Pleurocordyceps neoagarica, respectively.

Analyses 4 - Interspecific relationship of Ophiocordyceps

The combined dataset comprised 149 strains and/or specimens of Ophiocordyceps and included 4701 characters (including gaps), of which 1397 were parsimony-informative, 305 were singleton sites, and 2999 were constant. The aligned loci consisted of LSU (836 bp), ITS (510 bp), SSU (1,018 bp), tef-1α (900 bp), rpb1 (578 bp), and rpb2 (859 bp). Tolypocladium inflatum, T. ophioglossoides, and T. paradoxum were selected as outgroup taxa. The best-scoring maximum-likelihood (ML) tree, with a final log-likelihood value of −47,405.816, is shown in Fig. 6.

Fig. 6 — Maximum likelihood (IQ-TREE) phylogeny of *Ophiocordyceps* based on the combined LSU, ITS, SSU, *tef-1α*, *rpb1*, and *rpb2* sequence dataset. The newly obtained strains are indicated in blue text. SH-aLRT value (left) and the UFB value (right) equal to or greater than 50 and posterior probabilities (middle) equal to or higher than 0.90 are given near to the nodes. Superscript ‘T’ denotes the type strains/specimens.

Specimens HKAS 148572, HKAS 148571, and HKAS 148569 formed a clade sister to Hirsutella leigongshanensis with strong support (SH-aLRT = 94 %, PP = 1.00, UFB = 100 %; Fig. 6), and fall in the Ophiocordyceps pseudoacicularis complex.

Divergence time estimation

Molecular dating of the Hypocreales has been previously explored with utilization of the fossil Paleoophiocordyceps coccophagus to calibrate the Ophiocordyceps lineage (Sung et al. 2008, Wei et al. 2022, Yu et al. 2024). More recently, Zhuang et al. (2025) reported two entomopathogenic fungal fossils including Paleoophiocordyceps gerontoformicae sp. nov. and Paleoophiocordyceps ironomyiidarum (as “ironomyiae”) sp. nov., from mid-Cretaceous Kachin amber (approx. 99 MYA old). The authors inferred an Early Cretaceous origin (~130 MYA) for Ophiocordyceps based on updated fossil calibrations, a revision that pushes the estimated timeline back by approximately 30 MYA compared to earlier estimates. Building upon these advancements, the present study applied the updated fossil Paleoophiocordyceps gerontoformicae to calibrate the stem lineage of the zombie-ant fungi. To ensure robustness, the calibration points and representative taxa in this analysis were selected following Zhuang et al. (2025) by including species of Hirsutella clade, O. ravenelii clade and O. sphecocephala Clade within Ophiocordyceps.

Our molecular dating results indicate (Fig. 7) that the stem ages of Hypocreales families range from approximately 84 to 232 MYA (Table 1). This temporal span is largely consistent with the previously inferred Mesozoic diversification of major lineages within the order (Berbee & Taylor 2010, Hyde et al. 2017). Notably, the newly established family Chlorocilliaceae shared a common ancestor with Clavicipitaceae at approximately 142 MYA. This divergence falls well within the 50–150 MYA temporal window proposed for the establishment of family-level lineages in Sordariomycetes (Hyde et al. 2017). Furthermore, genera within this new family, such as Husseyia, Chlorocillium, Albacillium and Rousseaua exhibit comparable divergence depths. This pattern implies that these taxa diversified during a single evolutionary radiation event rather than representing disjointed, independently ancient relict lineages. Such temporal congruence provides strong evidence for their treatment as members of a monophyletic family, reflecting a shared evolutionary history.

Table 1.

Comparative stem-age estimates (MYA) of Hypocreales families inferred from different studies.

Family	a	b	c
Acremoniopsiaceae	155
Albomorchellophilaceae	89		150
Bionectriaceae	147	119	149
Calcarisporiaceae	89		150
Chlorocilliaceae	142
Chrysonectriaceae	134
Clavicipitaceae	142	129	147
Cocoonihabitaceae	132		133
Cordycipitaceae	162	151	174
Flammocladiellaceae	92	51	142
Hypocreaceae	113	151	157
Ljuhyaceae	92		128
Myrotheciomycetaceae	159		179
Nectriaceae	193	158	164
Neoacremoniaceae	115		136
Niessliaceae	134	200	160
Nothoacremoniaceae	134
Ophiocordycipitaceae	222	146	150
Polycephalomycetaceae	158	129	147
Pseudodiploösporaceae	113		157
Pseudoniessliaceae	168		133
Sarocladiaceae	232
Sedecimiellaceae	115
Stachybotryaceae	230	188	188
Stromatonectriaceae	84		103
Tilachlidiaceae	84	87	103
Valsonectriaceae	211		242
Xanthonectriaceae	119		128

Open in a new tab

Notes: a - this study, b - Wei et al. (2022), c - Yu et al. (2024).

Taxonomy

The taxonomy section treats a total of 18 genera and 30 species across six families: Calcarisporiaceae (one genus and three species), Chlorocilliaceae (nine genera and 16 species), Clavicipitaceae (one genus and one species), Cordycipitaceae (two genera and two species), Polycephalomycetaceae (four genera and seven species), and Ophiocordycipitaceae (one genus and one species). The content is hierarchically structured by family, genus, and species, documenting a wide array of taxonomic novelties including new species, new combinations, and new records. Notably, the section prioritizes detailed descriptions for the newly established family Chlorocilliaceae fam. nov., as well as the new genera Paralecanicillium gen. nov. and Neodingleyomyces gen. nov., while providing concise notes for all other taxa to clarify their classification and diversity.

Calcarisporiaceae Jing Z. Sun et al., Mycol. Prog. 16(4): 435. 2017. MB 817660.

Notes: The family Calcarisporiaceae was delineated by Sun et al. (2016) to encompass the monotypic genus Calcarisporium. The members of this family are fungicolous, thriving on other fungi (Sun et al. 2016, 2017, Zhu et al. 2023). Notably, Calcarisporium cordycipiticola is recognized as a mycoparasite, inflicting catastrophic diseases on the edible fungus Cordyceps militaris, and thereby posing a significant risk to its commercial cultivation (Liu & Dong 2023).

Calcarisporium Preuss, Linnaea 24: 124. 1851. MB 7465.

Members of Calcarisporium are characterized by erect, verticillately branched conidiophores bearing subulate conidiogenous cells with sympodially arranged, conidium-bearing denticles. Currently, the genus comprises eight species (MycoBank, accessed on 25 June 2025), namely Ca. acerosum (Matsushima 1975), Ca. arbuscula (Preuss 1851), Ca. cordycipiticola (Sun et al. 2016), Ca. guizhouense (Chen et al. 2024a), Ca. ovalisporum (Hoog 1978), Ca. phaeopodium (Somrithipol & Jones 2006), Ca. xylariicola (Sun et al. 2017), and Ca. yuanyangense (Zhu et al. 2023). Notably, the phylogenetic positions of Ca. acerosum, Ca. ovalisporum, and Ca. phaeopodium remain unresolved owing to the lack of available DNA sequence data (GenBank, accessed 25 June 2025). Most species of Calcarisporium are known as fungicolous taxa inhabiting the stromata of other fungi, including Cordyceps militaris, Ophiocordyceps nutans, and species of Xylaria (Sun et al. 2016, 2017, Zhu et al. 2023). In the present study, a novel species parasitizing Ophiocordyceps pseudolloydii and O. tricentri from China is introduced based on combined morphological and molecular evidence. Nevertheless, several ecological exceptions have been reported within the genus, including Ca. acerosum isolated from dead bark of Fagus crenata in Japan (Matsushima 1975), Ca. phaeopodium from decaying dicotyledonous leaves in Thailand (Somrithipol & Jones 2006), and Ca. ovalisporum from an insect host (Cercopoidea) in Sri Lanka (Hoog 1978). To date, species of Calcarisporium have been described predominantly on the basis of their asexual morphs, and their sexual morphologies have remained largely unknown. In this context, two new combinations possessing nectria-like sexual morphs are introduced in the present study, thereby expanding the morphological circumscription of Calcarisporium and providing the first insights into the sexual states of the genus.

Calcarisporium ophiocordycipiticola D.P. Wei & T.C. Wen, sp. nov. MB 859497. Fig. 8.

Etymology: The epithet “ophiocordycipiticola” refers to the hyperparasites inhabiting stromata of Ophiocordyceps species.

Description: Asexual morph: Colonies white, sparse, covering whole stromata and insect cadavers. Mycelium hyaline to pale brown, smooth-walled, immersed. Conidiophores 125–346 ( $\overline{x} = 252$ , n = 20) μm long, 5.2–5.8 ( $\overline{x} = 5.5$ , n = 20) μm wide near the base, 2.2–3.4 ( $\overline{x} = 2.8$ , n = 20) μm wide near the apex, conspicuous, solitary, macronematous, mononematous, straight or slightly flexuous, cylindrical, gradually tapering toward the apex, basal part thick-walled, 6–12-sepate, bifurcated at about 120 μm above the base or simple, becoming thin-walled toward the apex, with conidiogenous cells arranged in whorls every 11–24 um, 2–9 whorls and 1–5 conidiogenous cells in each whorl. Conidiogenous cells 8–16 × 1–3 ( $\overline{x} = 12 \times 1.8$ , n = 40) μm, subulate, smooth-walled, hyaline, gradually attenuated toward the apex, without a swollen apex, discrete, with sympodially arranged denticles (1–2 μm long) at tip. Conidia 2.3–5.8 × 1–4 ( $\overline{x} = 4.2 \times 1.8$ , n = 30) μm, holoblastic, unicellular, obclavate, hyaline, smooth, straight, thin-walled. Sexual morph: Undetermined.

Typus: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stroma of Ophiocordyceps pseudolloydii attached to a living dicotyledonous leaf, 24 Oct. 2023, D.P. Wei, MZH11 (holotype HKAS 148552).

Additional material examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stroma of Ophiocordyceps tricentri attached to a living dicotyledonous leaf, 24 Oct. 2023, D.P. Wei, DSSZ40 (HKAS 148551).

Notes: Multigene phylogenetic analyses recovered Calcarisporium ophiocordycipiticola, Neobaropsis andensis, and Neobarya agaricicola as a well-supported clade within Calcarisporiaceae (SH-aLRT = 88 %, PP = 0.96, UFB = 78 %; Fig. 3). Morphologically, Ca. ophiocordycipiticola differs from N. andensis, particularly in having subulate conidiogenous cells with denticulate apices, a character absent in the latter. Neobarya agaricicola exhibits an asexual morph resembling Calcarisporium. Its placement in Clavicipitaceae by Candoussau et al. (2007), based solely on ascus-cap morphology, has not been supported by molecular phylogenetic evidence. Morphologically, N. agaricicola can be distinguished from Ca. ophiocordycipiticola by its shorter conidiophores (50–130 μm vs 125–346 μm) and larger conidia (6.7–8.7 μm vs 2.3–5.8 μm). Notably, N. andensis and N. agaricicola share similar sexual characters, including superficial, brightly coloured perithecia, narrowly cylindrical asci with a thickened apical cap, and filiform ascospores. The close phylogenetic affinity of these taxa to Calcarisporium is inferred here for the first time. Based on combined phylogenetic evidence and morphological congruence, Neobaropsis andensis and Neobarya agaricicola are therefore transferred to Calcarisporium. As the sexual morph of Calcarisporium has not previously been documented, the inclusion of these two species provides the first insights into the sexual morph of the genus.

Calcarisporium andense (Flakus et al.) D.P. Wei & T.C. Wen, comb. nov. MB 861955.

Basionym: Neobaryopsis andensis Flakus et al., Pl. Fungal Syst. 64(2): 307. 2019.

Description: See Flakus et al. (2019).

Typus: Bolivia, Dept. Cochabamba, Pov. Tiraque, Parque, Nacional Carrasco, Camino de los Nubes, Antenas Sillar–Villa Tunari old road, 17°12’32” S, 65°41’52” W, 3520 m, upper montane Youngas cloud forest, corticolous, on Lobariella pallida, unknown collection date, A. Flakus, Adam Flakus 25967 (holotype in KRAM, isotype in LPB).

Notes: Neobaropsis andensis was originally placed in Cordycipitaceae based on its inferred phylogenetic affinity to Simplicillium (Flakus et al. 2019). However, multigene phylogenetic analyses in the present study, together with those of Darmostuk et al. (2025), consistently resolve Neobaropsis within Calcarisporiaceae. Neobaropsis andensis morphologically resembles Neobarya agaricicola in producing nectria-like perithecia, narrowly cylindrical asci with a distinct apical cap, and filiform ascospores, but differs from the latter in having conidiogenous cells that lack apical denticles. Neobarya agaricicola exhibits calcarisporium-like conidiogenous cells, indicating that it belongs to Calcarisporium (Candoussau et al. 2007). Neobaropsis andensis is therefore transferred to Calcarisporium as Calcarisporium andense, based on its close phylogenetic relationship with N. agaricicola and Calcarisporium.

Calcarisporium agaricicola (Berk.) D.P. Wei & T.C. Wen, comb. nov. MB 861956.

Basionym: Nectria agaricicola Berk. in Hooker, Botany of the Antarctic Voyage. iii. Flora Tasmaniae 2: 278. 1859.

Synonyms: Ophionectria agaricicola (Berk.) Sacc., Syll. Fung. 2: 563. 1883.

Barya agaricola (Berk.) Höhnel, Ann. Mycol. 8: 464. 1910.

Neobarya agaricicola (Berk.) Samuels & Lowen, Sydowia 59(2): 185. 2007.

Description: See Candoussau et al. (2007)

Typus: On decaying Agaricus mushrooms in Tasmania.

Notes: Neobarya agaricicola is a mycoparasitic species characterized by its superficial, brightly coloured perithecia produced abundantly on the cap and stipe of small marasmioid agarics. Since its original recognition, the taxonomic placement of this species has been primarily based on its unique sexual morph morphology, including elongate-ovoidal perithecia with a thick-walled, two-layered perithecial wall, asci with a conspicuous apical cap, and filiform, aseptate ascospores (Candoussau et al. 2007). The asexual morph of N. agaricicola was known as calcarisporium-like based on the culture made from germinating asci of specimen PDD 50035. This asexual morph is characterized by verticillately branched conidiophores, tapering conidiogenous cells bearing terminal clusters of denticles, and cylindrical to slightly clavate conidia held in dry heads (Candoussau et al. 2007). These characters are consistent with the generic concept of Calcarisporium. Accordingly, Neobarya agaricicola is transferred to Calcarisporium based on phylogenetic evidence and characteristics of the asexual morph.

Chlorocilliaceae D.P. Wei & T.C. Wen, fam. nov. MB 859498.

Etymology: Named after the type genus, Chlorocillium.

Type genus: Chlorocillium Zare & W. Gams, Mycol. Prog. 15: 1005. 2016

Description: Entomopathogenic or fungicolous or saprobic; Sexual morph: Stromata absent. Perithecia directly arising from loose mycelial mat, co-occurring with the asexual morph, superficial, pale yellow, ovoid, ostiolate. Asci filiform, with a thickened apex. Ascospore hyaline, aseptate, filiform, intact at maturity. Asexual morph: Colonies on natural substrate white to pale green, fluffy to powdery. Synnemata comprised of loose hyphae, cylindrical, erect, cespitose. Conidiophores micronematous, branched, laterally or apically producing solitary to cluster of phialides. Phialides flask-shaped, subulate, typically tapering toward apex from middle part, straight, curved or zigzag, smooth-walled, 0–2-septate, apically branched or unbranched. Conidia hyaline, aseptate, crescent, fusiform, oblong, obovate, with or without a scar at the end, usually gathering in chains or slimy head.

Notes: The multigene-based phylogenetic analysis showed that Albacillium, Chlorocillium, Fiorinimazzantia, Husseyia, Neoaraneomyces, Paraneoaraneomyces, Subuliphorum, Rousseaua, and Speluncomyces formed distinct lineages sister to the Clavicipitaceae. Three main subclades were observed within this lineage. The first clade includes a single genus, Chlorocillium. The second comprises Albacillium, Rousseaua, Speluncomyces, Husseyia, and Fiorinimazzantia. The third consists of Paraneoaraneomyces, Subuliphorum, and Neoaraneomyces. These subclades correspond to notable differences in conidial morphology and arrangement. Species with conidia arranged in chains are generally found in the first subclade, while those with conidia arranged in globose heads dominate the second and third subclades. Neoaraneomyces is a notable exception, containing species with both types of conidial arrangement (see Fig. 3). This phylogenetic framework provides a clearer understanding of the evolutionary relationships and morphological distinctions within these genera. However, despite these new insights, the taxonomic placement of several genera has remained controversial. Fiorinimazzantia, Husseyia, and Rousseaua have historically been considered as Sordariomycetes incertae sedis (Tan & Shivas 2023a, b), while the other six genera were previously classified within Clavicipitaceae (Chen et al. 2019, 2022, 2024a, 2025, Zhang et al. 2023, Dong et al. 2025, Lin et al. 2025). The phylogenetic relationships revealed in this study provide the first comprehensive clarification of their positions, leading us to propose a new family, Chlorocilliaceae, to accommodate this distinct lineage. This new family includes 33 species and has a broad host range across three kingdoms (Animalia, Plantae, and Fungi), with a particular preference for spiders and the stromata or synnemata of entomopathogenic fungi (Table 2). The generic boundaries within this family have now been more clearly defined, providing a stronger foundation for future studies on these genera.

Table 2.

Synopsis of species of Chlorocilliaceae.

Species	Specimen or culture	Host / sources	Phialides (μm)	Conidia (μm)	References
Albacillium fuzhouense	CGMCC 3.27818	Larva of Coleoptera	Lanceolate, 13.6–81.5 × 0.8–1.3	Fusiform, ellipsoidal, slightly curved, 2.4–6.7 × 1.0–1.4	Lin et al. (2025)
Albacillium hingganense	SGSF 339 / HKAS 135603	Leaf litter / Ophiocordyceps cf. unilateralis	Short to slender, tapering toward the apex, 8.6–60.1 × 0.9–1.5 / cylindrical, tapering toward the apex, 10–28 long, base 0.7–1.6 wide, tip 0.4–0.9 wide	Ellipsoidal to cymbiform, straight to slightly curved, 2.0–4.3 × 0.8–1.3 / reniform, 1.6–2.4 × 0.8–1.2	Ding et al. (2024), this study
Chlorocillium aschersoniae.	HKAS 135599.	Aschersonia cf. samoensis	flask-shaped, apically tapering into a narrow neck, 9–17 long, base 1.3–2.4 wide, tip 0.6–0.9 wide	fusiform, 2.7–4.5 × 1.2–2.0	This study
Chlorocillium araneogenum	DY101801	Spider	Cylindrical, tapering to a thin neck, 8.3–23.3 × 1.3–2.2	Fusiform, 3.4–5.8 × 1.4–1.8	Chen et al. (2024a)
Chlorocillium coccidiicola	HKAS 135610	Scale insect	Cylindrical with thin neck, 6.4–12 long, base 1–2 wide, tip 0.4–0.6 wide	Broadly fusiform, 2.3–3.6 × 1.1–1.8	This study
Chlorocillium cordycipiticola	HKAS 135604	Cordyceps cf. neopruinosa	Subulate, 12–39 long, base 1–1.6 wide, tip 0.4–0.8 wide	ovoid, 1.7–3 × 0.8–1.3	This study
Chlorocillium duiganiae	MST F29447	Dead Insecta	n/a	n/a	Tan et al. (2025)
Chlorocillium gibellulae	HKAS 135594	Gibellula sp.	Subulate, 10–39 long, base 1.1–1.7 wide, tip 0.5–1 wide	Fusiform to obovate, 3–5 × 1.2–2	This study
Chlorocillium globiceps	HKAS 135618	Ophiocordyceps cf. globiceps	Flask-shaped, 9–17 long, base 1.2–2.5 wide, tip 0.5–1.1 wide	Fusiform, 3–4.5 × 1.4–2.2	This study
Chlorocillium griseum	CBS 387.73	Aphid	Lanceolate, tapering gradually from the middle to apex, 18–40(−55) × 2–2.5	Fusiform, 4.5–6 × 1.0–1.5	Zare & Gams (2016)
Chlorocillium gueriniae	BRIP 72680a	Spider	n/a	n/a	Tan & Shivas (2023a)
Chlorocillium guizhouense	DL10171 / HKAS 148578	Spider	Cylindrical at basal portion, tapering into a distinct neck, 11.6–25.3 × 1.0–1.2 μm / flask-shaped, 9.7–20 long, base 1.1–2.6 wide, tip 0.6–0.9 wide	Fusiform, 2.6–3.8 × 1.1–1.6 / fusiform, 3–4.2 × 1–1.6	Chen et al. (2025a), this study
Chlorocillium houqiaoense	HKAS 148564	Spider	Subulate, 5.2–19 long, base 0.9–1.4 wide, tip 0.5–0.9 wide	Fusiform to obovate, 1.5–4 × 0.6–1.6	This study
Chlorocillium lepidopterorum	SD05361	Lepidopteran pupa	Cylindrical, tapering to a thin neck, 21.2–33.7 × 1.1–1.4	Fusiform, 3.1–4.3 × 1.3–1.5	Chen et al. (2024a)
Chlorocillium mauryae	MST F26633	Spider	n/a	n/a	Tan et al. (2024)
Chlorocillium montefioreae	BRIP 70299a	Spider	n/a	n/a	Tan & Shivas (2024)
Chlorocillium neolepidopterorum	GZAC DL1022	Lepidopteran pupa	Cylindrical, tapering to a thin neck, 7.3–14.4 × 1.0–1.7	Fusiform, 2.3–3.1 × 1.0–1.4	Chen et al. (2019), this study
Chlorocillium menghaiense	HKAS 135597 / HKAS 148570 / HKAS 135600	Ophiocordyceps nutans / Ophiocordyceps shimenxiaensis / Cordyceps militaris	Flask-shaped, 8–18 long, base 1.3–2.8 wide, tip 0.6–1.1 wide	Ovoid, 3.1–4.7 × 1.2–2.2	This study
Chlorocillium puerense	HKAS 135606	Hevansia-like fungus	Subulate with zigzag at upper part, 13–47 long, base 0.8–1.4 wide, tip 0.3–0.7 wide	Oblong, 2–3 × 0.8–1.3	This study
Chlorocillium rongjiangense	DL10301	Lepidopteran pupa	Cylindrical at the base, tapering into a thin neck, 19.7–34.3 × 0.7–1.1	Subwedge, 2.5–3.5 × 1.0–1.1	Chen et al. (2019)
Chlorocillium sanduense	HKAS 135608	Ascopolyporus sanduensis	Subulate,14–45 long, base 1.1–1.8 wide, tip 0.4–0.8 wide	Fusiform, 2.5–3.7 × 1–1.5	This study
Chlorocillium sinense	KY07181	Spider	Cylindrical, tapering to a thin neck, 11.7–20.1 × 1.1–1.3	Fusiform to ellipsoidal, 1.9–2.9 × 0.8–1.2	Chen et al. (2024a)
Chlorocillium taiyangheense	HKAS 135602 / HKAS 148568	Spider / Ophiocordyceps cf. oxycephala	Flask-shaped, 7.6–18 long, base 1–2 wide, tip 0.4–0.8 wide	Fusiform, 2.4–3.8 × 0.8–1.3	This study
Chlorocillium tengchongense	HKAS 148567	Lepidopteran larvae	Subulate, 25–79 long, base 2.6–4.8 wide, tip 1.6–2.6 wide	Ovoid, 3.8–10.5 × 1.7–4	This study
Chlorocillium vallense	DY09021	Spider	Lanceolate, tapering gradually from the middle to apex, 9.2–17.1 × 0.8–1.2	Fusiform, 2.0–3.2 × 0.9–1.4	Chen et al. (2025a)
Chlorocillium winlockiae	MST F3581	Dead Insecta	n/a	n/a	Tan et al. (2024)
Fiorinimazzantia elisabettae	BRIP 72660a / HKAS 135605	Decayed insect colonized by an unidentified hirsutella-like fungus / Hevansia minuta	Lanceolate, 20–36 long, base 0.8–1.6 wide, tip 0.4–0.7 wide	Allantoid, 2.3–3.6 × 0.6–1.4	Tan & Shivas (2023a), this study
Husseyia annamariae	BRIP 72654a	Decayed spider colonised by an unidentified hevansia-like fungus	n/a	n/a	Tan & Shivas (2023a)
Husseyia queenslandica	BRIP 72669a	Dead Arthropoda	n/a	n/a	Tan & Shivas (2023b)
Husseyia ramosissima	HKAS 135601	Akanthomyces sp.	Subulate, 13–35 long, base 0.7–1.6 wide, tip 0.3–0.6 wide	Reniform, 2.2–3.4 × 0.6–1.2	This study
Neoaraneomyces araneicola	DY101711	Spider	Lanceolate, 8.9–23.8 × 1.1–1.6	Fusiform to ellipsoidal, 2.9–4.4 × 1.3–2.0	Chen et al. (2022)
Neoaraneomyces wuyishanensis	CGMCC 3.28307	Spider	Lanceolate, 6.2–34.3 × 0.9–2.0	Ellipsoidal, fusiform, slightly curved, 2.5–4.5 × 1.0–1.4	Lin et al. (2025)
Paraneoaraneomyces sinensis	ZY 22.006	The green belt soil	Lanceolate, 19.0–34.0 × 0.5–1.5	Cymbiform to reniform, 3.0–5.5 × 1.0–1.5	Zhang et al. (2023)
Rousseaua marietteae	BRIP 62733a	Dead Sternorrhyncha	n/a	n/a	Tan & Shivas (2023b)
Speluncomyces lunatus	COAD 3666	On leaf litter from a ferruginous cave	Cylindrical to subulate, 7.5–25.5 × 0.8–1.5	Allantoid to lunate, 2–7.5 × 1–1.5	Dong et al. (2025)

Open in a new tab

Chlorocillium Zare & W. Gams, Mycol. Prog. 15: 1005. 2016. MB 815506.

Synonym: Pseudometarhizium W.H. Chen et al., MycoKeys 91: 59. 2022.

Diagnosis: Differs from other genera of Chlorocilliaceae in producing conidia aggregated in chains.

Description: Sexual morph: Perithecia sparsely produced on the mycelial mat covering the stromata or colonies of fungal hosts, superficial on loose mycelia, ovoid, ostiolate, pale yellow. Asci filiform, hyaline, unitunicate, with a thickened apex. Ascospore hyaline, filiform, non-disarticulating, aseptate, smooth-walled. Asexual morph: Colonies on natural substrate white, green to yellow green, fluffy to powdery. Synnemata arising from colonies, cylindrical, unbranched or branched, cespitose, comprised of loose, hyaline, septate hyphae. Conidiophores emerging from aerial hyphae or chondroid mycelia, bearing solitary, verticillate or crowded phialides. Phialides phialidic, slender, subulate to flask-shaped, straight or curved, gradually tapering from middle part. Conidia hyaline, aseptate, fusiform, ovoid, cylindrical, held in chains.

Type species: Chlorocillium griseum (Petch) Zare & W. Gams, Mycol. Prog. 15: 1007. 2016.

Basionym: Acremonium griseum Petch, Trans. Brit. Mycol. Soc. 11 (3–4): 262. 1926.

Synonyms: Verticillium griseum (Petch) W. Gams, Cephalosporiumartige Schimmelpilze (Stuttgart): 191. 1971.

Paecilomyces griseus (Petch) Subram., Kavaka 5: 98. 1979.

Notes: Acremonium griseum, the basionym of Ch. griseum, was found to parasitize abortive synnemata of Hirsutella, rather than insects (Petch 1926). The holotype of Acremonium griseum was collected by Petch from Hakgala, Sri Lanka and deposited in the herbarium K(M). Meanwhile, other collections from spiders from the same island and one from Pseudococcus sp. (scale insect) from Mauritius have been identified as Ac. griseum based on morphological characteristics zare & Gams 2016. Gams (1971) examined these specimens and concluded that Ac. griseum should be a synonym of Verticillium griseum, which was later renamed as Paecilomyces griseus by Subramanian (1978). Zare & Gams (2016) have designated the specimen CBS H-19258 (dried material of living culture CBS 387.73) as the epitype for Ch. griseum to link it with a sequenced culture and provided micromorphological illustrations for this species. Subsequently, nine new taxa from China and Australia have been added to the genus, including six araneopathogenic species (Chlorocillium araneogenum, Ch. guizhouense, Ch. mauryae, Ch. montefioreae, Ch. sinense, Ch. vallense), two species (Ch. gueriniae and Ch. winlockiae) found on an unidentified arthropod, and one species (i.e. Ch. lepidopterorum) associated with a lepidopteran larva (Chen et al. 2022, 2024a, Tan & Shivas 2023a, 2024). It is worth noting that the morphological illustrations for species native to Australia are absent from the respective protologues (Tan & Shivas 2023a, 2024). This omission underscores the need for a re-examination of their holotypes to elucidate their morphological attributes with greater precision. Historically, Chlorocillium has been categorized as Hypocreales genus incertae sedis (Wijayawardene et al. 2018, Hyde et al. 2024). However, a revision by Chen et al. (2024a) accommodated the genus in the Clavicipitaceae on the basis of multigene phylogenetic analyses. In the present study, we propose Chlorocilliaceae, a new family to accommodate Chlorocillium and eight other genera (including Albacillium, Fiorinimazzantia, Husseyia, Neoaraneomyces, Paraneoaraneomyces, Rousseaua, Speluncomyces and Subuliphorum), as they form a monophyletic clade that is separated from Clavicipitaceae. This taxonomic revision is accompanied by the addition of eleven new species.

Chlorocillium aschersoniae D.P. Wei & T.C. Wen, sp. nov. MB 859501. Fig. 9.

Etymology: The epithet “aschersoniae” is named after the genus of its host fungus, Aschersonia.

Description: Hyperparasitic on Aschersonia cf. samoensis. Asexual morph: Synnemata white, multiple, scattered, erect, cylindrical, comprised of hyaline, smooth-walled hyphae that are parallelly arranged. Conidiophores developing from superficial hyphae of synnemata, micronematous, branched, septate, terminating in cluster of phialides, densely compact. Phialides 9–17 ( $\overline{x} = 12$ , n = 30) μm long, base 1.3–2.4 ( $\overline{x} = 1.9$ , n = 30) μm wide, tip 0.6–0.9 ( $\overline{x} = 0.8$ , n = 30) μm wide, monophialidic, flask-shaped, apically tapering into a narrow neck, hyaline, smooth-walled, aseptate, straight or curved. Conidia 2.7–4.5 × 1.2–2.0 ( $\overline{x} = 3.5 \times 1.4$ , n = 30) μm, hyaline, fusiform, aseptate, smooth-walled, gathering in short chains. Sexual morph: Undetermined.

Typus: China, Yunnan Province, Xishuangbanna Tropical Botanical Garden, on stroma of Aschersonia cf. samoensis attached to underside of living leaves of Caryota sp., 12 Sep. 2023, D.P. Wei, XTBG08 (holotype HKAS 135599).

Additional material examined: China, Yunnan Province, Xishuangbanna Tropical Botanical Garden, on stroma of Aschersonia cf. samoensis attached to underside of living leaves of Caryota sp., 12 Sep. 2023, D.P. Wei, XTBG08a (HKAS 148559).

Notes: Chlorocillium aschersoniae constitutes a distinct clade that is sister to Ch. guizhouense. Although Ch. aschersoniae is genetically very closely related to Ch. guizhouense, these two species exhibit significant differences in their colonial characteristics. Chlorocillium aschersoniae develops white, fluffy colonies on the stroma of Aschersonia cf. samoensis, from which solitary, erect, sparse synnemata emerge. In contrast, Ch. guizhouense is an obligate pathogen of spider hosts and forms compact, pale green, powdery colonies that entirely envelop the spider hosts, giving rise to numerous cylindrical synnemata. Furthermore, the PHI test of Ch. aschersoniae showed no significant recombination (Φw = 0.2108) with their closely related taxa (Ch. Guizhouense, Ch. globiceps, and Ch. taiyangheense) (Fig. 4C1). Therefore, we propose Ch. aschersoniae as a separate species based on its pronounced differences in colonial characteristics and its respective associated hosts.

Chlorocillium coccidiicola D.P. Wei & T.C. Wen, sp. nov. MB 859502. Fig. 10.

Etymology: The epithet “coccidiicola” means that this fungus colonizes scale insects.

Description: Parasitic on scale insect. Asexual morph: Sporodochia white, pulvinate, powdery, entirely covering the host. Conidiophores micronematous, branched, septate, penicillate, bearing cluster of phialides, densely aggregated, forming a hymenium. Phialides 6.4–12.0 ( $\overline{x} = 9$ , n = 40) μm long, base 1–2 ( $\overline{x} = 1.6$ , n = 40) μm wide, tip 0.4–0.6 ( $\overline{x} = 0.5$ , n = 35) μm wide, monophialidic, cylindrical, apically narrowing into a thin neck, hyaline, smooth-walled, aseptate, straight or curved. Conidia 2.3–3.6 × 1.1–1.8 ( $\overline{x} = 2.8 \times 1.4$ , n = 50) μm, hyaline, broadly fusiform, aseptate, smooth-walled, gathering in chains. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 13 mm diam. after incubation for 60 d at 25 °C, white (1A1), velvety, nearly circular, umbonate, surface smooth, mycelia moderately dense, edge undulate, reverse curry yellow (4C8) at central zone, becoming paler toward the margin.

Typus: China, Guizhou Province, Qianxinan Buyei and Miao Autonomous Prefecture, Anlong County, Xianheping National Nature Reserve, on scale insect attached to a living stem of herbaceous plant, 6 Sep. 2023, Z.L. Liu, XHP2023090741L (holotype HKAS 135610, culture ex-type KUNCC24-17900).

Additional materials examined: China, Guizhou Province, Qianxinan Buyei and Miao Autonomous Prefecture, Anlong County, Xianheping National Nature Reserve, on scale insect attached to a living stem of herbaceous plant, 6 Sep. 2023, S.W. Xie, AL20230907X (HKAS 135609, living culture KUNCC24-17901).

Notes: Chlorocillium coccidiicola forms a distinct clade in Chlorocillium (Figs 3, 4). This species is characterized by its straight, symmetrical phialides, which are cylindrical at the base and abruptly narrow into a short apex (Fig. 10G–J). In contrast, other Chlorocillium species typically produce slightly curved, asymmetric phialides that gradually narrow from the middle portion. Hence, based on morpho-molecular analyses, we introduce Chlorocillium coccidiicola as a new species.

Chlorocillium cordycipiticola D.P. Wei, Z. L. Liu & T.C. Wen, sp. nov. MB 859504. Fig. 11.

Etymology: The epithet “cordycipiticola” means this fungus parasitizes Cordyceps species.

Description: Hyperparasitic on Cordyceps cf. neopruinosa growing on a pupa of Limacodidae. Stromata of fungal host multiple, cylindrical, orange, fleshy. Asexual morph: Colonies entirely covering the stromata of C. cf. neopruinosa, white, fluffy. Conidiophores differentiating from interlaced hyphae, micronematous, branched, bearing solitary, biverticillate to verticillate phialides. Phialides 12–39 ( $\overline{x} = 21$ , n = 25) μm long, base 1.0–1.6 ( $\overline{x} = 1.3$ , n = 25) μm wide, tip 0.4–0.8 ( $\overline{x} = 0.6$ , n = 25) μm wide, monophialidic, smooth-walled, straight to curved, subulate, gradually tapering toward the apex. Conidia 1.7–3.0 × 0.8–1.3 ( $\overline{x} = 2.2 \times 1.0$ , n = 45) μm, hyaline, ovoid with one pointed end, aseptate, smooth-walled, gathering in chains. Sexual morph: Perithecia 177–426 × 97–214 ( $\overline{x} = 306 \times 156$ , n = 10) μm, superficial, pale yellow, ovoid, ostiolate. Asci 171–210 × 2.2–4.0 ( $\overline{x} = 187 \times 3.0$ , n = 15) μm, filiform, with a thickened apex. Ascospores 120–145 × 0.7–1.2 ( $\overline{x} = 136 \times 1.0$ , n = 10) μm, hyaline, aseptate, filiform, intact when mature.

Culture characteristics: Colonies on PDA attaining 35 mm diam. after incubation for 60 d at 25 °C, white (1A1), velvety, circular, umbonate, mycelia moderately dense, surface roughed with radial wrinkles at marginal zone, edge lobate, reverse agate (7E8) at central zone, white at margin.

Typus: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stromata of Cordyceps cf. neopruinosa immersed in soil, 25 Oct. 2023, Z.L. Liu, XF20231027154L (holotype HKAS 135604, living culture ex-type KUNCC24-17918).

Additional material examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stromata of Cordyceps cf. neopruinosa immersed in soil, 25 Oct. 2023, Z.L. Liu, 154Lb (HKAS 148548).

Notes: Chlorocillium cordycipiticola is phylogenetically closely related to Ch. neorongjiangense and Ch. tengchongense (Fig. 4). However, Chlorocillium cordycipiticola differs from Ch. neorongjiangense in that it is a hyperparasite on Cordyceps cf. neopruinosa, forming velvety colonies that entirely cover the stroma of the host fungus, whereas Ch. neorongjiangense is an entomopathogen that infects lepidopteran pupae and produces robust synnemata from which white colonies emerge (Chen et al. 2019). Chlorocillium tengchongense is distinguished from Ch. cordycipiticola by its obligate growth on the hairs of lepidopteran insects and its larger conidia (3.8–10.5 × 1.7–4.0 μm vs 1.7–3.0 × 0.8–1.3 μm). A comparison of ITS, LSU, tef-1α, and rpb1 nucleotide sequences reveals no significant differences between Ch. cordycipiticola, Ch. neorongjiangense, and Ch. tengchongense. However, an analysis of rpb2 sequence data shows that Ch. cordycipiticola differs from Ch. neorongjiangense and Ch. tengchongense by 17/1040 bp (1.6 %) and 14/908 bp (1.6 %), respectively. Moreover, the result of PHI test was 1.0 indicating that there is no significant recombination between Ch. cordycipiticola and its closely related species (Fig. 4C4).

Chlorocillium gibellulae D.P. Wei & T.C. Wen, sp. nov. MB 859505. Fig. 12.

Etymology: Named after the generic name of the host fungus Gibellula sp.

Description: Hyperparasitic on Gibellula sp. Asexual morph: Colonies white, fluffy, entirely covering synnemata of Gibellula sp. Mycelium 1.0–2.0 ( $\overline{x} = 1.4$ , n = 30) μm wide, hyaline, branched, smooth-, thick-walled, rarely septate. Conidiophores emerging from anastomosing mycelia, erect, bearing solitary or biverticillate phialides at each node. Phialides 10–39 ( $\overline{x} = 24$ , n = 35) μm long, base 1.1–1.7 ( $\overline{x} = 1.4$ , n = 35) μm wide, tip 0.5–1.0 ( $\overline{x} = 0.8$ , n = 35) μm wide, monophialidic, slender, subulate tapering gradually from the middle to apex, smooth-walled, aseptate, straight or curved. Conidia 3–5 × 1.2–2.0 ( $\overline{x} = 4 \times 1.6$ , n = 40) μm, hyaline, fusiform to obovate with scar at one end, aseptate, smooth-walled, catenulate. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 14 mm diam. after incubation for 60 d at 25 °C, white (1A1), felty, circular, mycelia dense, edge entire, surface lumpy with radially striated, reverse reddish ochre (7D8).

Typus: China, Yunnan Province, Menghai County, Gelanghe village, on synnemata of Gibellula sp. parasitic on spider attached to underside of living leaf of Fagaceae, 12 Sep. 2022, D.P. Wei, GLH16 (holotype HKAS 135594).

Additional material examined: China, Yunnan Province, Menghai County, Gelanghe village, on synnemata of Gibellula sp. parasitic on spider attached to underside of living leaf of Fagaceae, 12 Sep. 2022, D.P. Wei, GLH10 (HKAS 135595, living culture KUNCC24-17902).

Notes: The IQ-TREE phylogeny (Fig. 4) showed that Chlorocillium gibellulae along with Ch. mauryae and Ch. houqiaoense form a distinct lineage within the genus Chlorocillium. Chlorocillium gibellulae is distinguished by its hyperparasitic lifestyle on the synnemata of Gibellula species, contrasting with other members of this clade, which are pathogens of spiders. The protologue of Ch. mauryae lacks a detailed morphological description, preventing direct morphological comparisons (Tan et al. 2024). Although Ch. gibellulae shows no significant genetic divergence from Ch. mauryae in the ITS, LSU, and tef-1α regions, it exhibits a 1.9 % difference (17/892 bp) in the rpb2 sequences. Morphologically, Ch. gibellulae can be differentiated from Ch. houqiaoense by its larger phialides (10–39 × 1.1–1.7 μm vs 5.2–19 × 0.9–1.4 μm) and conidia (3–5 × 1.2–2.0 μm vs 1.5–4.0 × 0.6–1.6 μm), as well as the absence of synnemata. Additionally, the PHI test of Ch. gibellulae showed no significant recombination (Φw = 1.0) with their closely related taxa (Fig. 4C3). Hence, we determined that our specimens are a novel species of Chlorocillium.

Chlorocillium globiceps D.P. Wei & T.C. Wen, sp. nov. MB 859506. Fig. 13.

Etymology: Named after the epithet of its host fungus, Ophiocordyceps cf. globiceps.

Description: Hyperparasitic on Ophiocordyceps cf. globiceps. Asexual morph: Synnemata pale yellow-green, fluffy to powdery, cylindrical, erect, cespitose, comprising hyaline, septate hyphae that are parallel-arranged. Conidiophores emerging from superficial hyphae of synnemata, micronematous, branched, septate, bearing cluster of phialides. Phialides 9–17 ( $\overline{x} = 11$ , n = 40) μm long, base 1.2–2.5 ( $\overline{x} = 2$ , n = 40) μm wide, tip 0.5–1.1 ( $\overline{x} = 0.8$ , n = 35) μm wide, monophialidic, flask-shaped, narrowing into a thin neck, hyaline, smooth-walled, aseptate, straight or curved. Conidia 3–4.5 × 1.4–2.2 ( $\overline{x} = 4 \times 1.7$ , n = 35) μm, hyaline, fusiform with scar at one end, aseptate, smooth-walled, gathering in short chains. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 25 mm diam. after incubation for 60 d at 25 °C, white (1A1), circular, velvety, convex, mycelia filamentous, aerial, surface smooth, edge lobate, reverse ginger (5D6).

Typus: China, Jilin Province, Baishan City, Fusong County, Shuihe Town, on stroma of Ophiocordyceps cf. globiceps attached to tree bark, 24 Aug. 2023, T.C. Wen & Z.L. Liu, H14 (holotype HKAS 135618, culture ex-type KUNCC24-17911).

Additional materials examined: China, Jilin Province, Baishan City, Fusong County, Shuihe Town, on stroma of Ophiocordyceps cf. globiceps attached to tree bark, 24 Aug. 2023, T.C. Wen & Z.L. Liu, H18 (HKAS 135619, living culture KUNCC24-17912); ibid., H29 (HKAS 135621, living culture KUNCC24-17909); ibid., H32 (HKAS 135622, living culture KUNCC24-17913); ibid., H43 (HKAS 135616, living culture KUNCC24-17910).

Notes: Chlorocillium globiceps is accommodated in a sister clade to Ch. taiyangheense (Fig. 4). Chlorocillium globiceps exhibits high specificity to the host fungus, Ophiocordyceps cf. globiceps, while Ch. taiyangheense is a generalist, occurring on spiders and stromata of Ophiocordyceps cf. oxycephala. Additionally, Ch. globiceps can be easily distinguished from Ch. taiyangheense by its pale green, powdery, cylindrical, erect synnemata on the natural hosts, while the latter species produces white, velvety synnemata or colonies on its host. The result of the PHI test of Ch. globiceps and its closely related species was 0.2108 (Fig. 4C1) and revealed that those species were different.

Chlorocillium guizhouense W.H. Chen et al., MycoKeys 112: 346. 2025. MB 856175. Fig. 14.

Description: Parasitic on a spider. Asexual morph: Colonies on spider, pale green, powdery, compact. Synnemata cylindrical, arising from colonies covering spider hosts, pale green, powdery, consisting of parallel-arranged, hyaline, thin-walled, septate hyphae. Conidiophores developing from hyphae of synnemata and colonies, micronematous, hyaline, irregularly branched, laterally or apically bearing solitary or cluster of phialides. Phialides 9.7–20.0 ( $\overline{x} = 13$ , n = 30) μm long, base 1.1–2.6 ( $\overline{x} = 1.6$ , n = 30) μm wide, tip 0.6–0.9 ( $\overline{x} = 0.7$ , n = 30) μm wide, monophialidic, flask-shaped with cylindrical base and narrow neck, smooth-walled, aseptate, commonly curved. Conidia 3.0–4.2 × 1.0–1.6 ( $\overline{x} = 3.6 \times 1.4$ , n = 30) μm, hyaline, fusiform, aseptate, smooth-walled, catenulate. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 28 mm diam. after incubation for 60 d at 25 °C, initially white (1A1), velvety, circular, mycelia dense, edge entire, slightly raised, reverse reddish ochre (7D8), at centre, apricot (5B6) at margin, producing pale brick red (7D7) pigment diffusing into agar.

Typus: China, Guizhou Province, Qiandongnan Miao and Dong Autonomous Prefecture, Rongjiang County, Dali Dong Village, on a dead spider (Araneae), 1 Oct. 2018, W. Chen (holotype GZAC DL1017, culture ex-type DL10171).

Materials examined: China, Jilin Province, Baishan City, Fusong County, Shuihe Town, on spider attached to bark of living tree, 24 Aug. 2023, T.C. Wen et al., H79 (HKAS 148578); ibid., H20 (HKAS 135620, living culture KUNCC24-17905); ibid., H65 (HKAS 135624, living culture KUNCC24-17906); ibid., H48 (HKAS 135592, living culture KUNCC24-17903); ibid., H59 (HKAS 135617, living culture KUNCC24-17908); ibid., H57 (HKAS 135623, living culture KUNCC24-17907); ibid., H58 (HKAS 135615, living culture KUNCC24-17904); ibid., H171 (HKAS 148582); ibid., H169 (HKAS 148581, living culture KUNCC25-19463); ibid., H129 (HKAS 148579, living culture KUNCC25-19464,); ibid., H71C (HKAS 148577, living culture KUNCC25-19465); ibid., H183 (HKAS 148583, living culture KUNCC25-19466); ibid., H146 (HKAS 148580, living culture KUNCC25-19467).

Notes: Chlorocillium guizhouense was introduced by Chen et al. (2025) from a dead spider in Rongjiang County, Guizhou Province, the southern part of China. In this study, we collected many specimens of this species from northern China. Multigene phylogenetic analyses reveal that our strains group with the ex-type of Ch. guizhouense with adequate support (Fig. 4). Additionally, a comparison of nucleotide sequences shows that there are no significant differences between our strain and the ex-type strain in ITS, LSU, tef-1α, and rpb2 regions, indicating that they are conspecific. This species is obligately pathogenic on spiders. However, the culture characteristics derived from natural hosts are different between specimens from southern and northern China. Our strains are characterized by yellow-green, powdery, compact colonies, while those of the type are white, velvety, and sparse.

Chlorocillium houqiaoense D.P. Wei, Q. F. Huang & T.C. Wen, sp. nov. MB 859507. Fig. 15.

Etymology: Named after the collection site of the type specimen, Houqiao Town in Tengchong City of Yunnan Province.

Description: Parasitic on a spider. Asexual morph: Colonies on spider white, fluffy, sparse. Vegetative mycelium 1.0–2.4 ( $\overline{x} = 1.6$ , n = 25) μm wide, hyaline, branched, smooth-walled, aseptate. Synnemata arising from the legs of spider, white, velvety, tufted, consisting of anastomosing hyphae along with crowded conidiophores. Conidiophores developing from superficial hyphae of synnemata, irregularly branched, typically bearing solitary or biverticillate phialides at each node. Phialides 5.2–19 ( $\overline{x} = 13$ , n = 40) μm long, base 0.9–1.4 ( $\overline{x} = 1.2$ , n = 40) μm wide, tip 0.5–0.9 ( $\overline{x} = 0.7$ , n = 40) μm wide, monophialidic, subulate tapering gradually from the middle to apex, smooth-walled, aseptate, straight or curved. Conidia 1.5–4.0 × 0.6–1.6 ( $\overline{x} = 2.5 \times 1.0$ , n = 40) μm, hyaline, fusiform to obovate with scar at one end, aseptate, smooth-walled, catenulate. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 20 mm diam. after incubation for 60 d at 25 °C, white (1A1) at centre, regency green (30B3) at periphery, floccose, nearly circular, mycelia loose, surface pleated, annular, edge entire, reverse brick red (7D7).

Typus: China, Yunnan Province, Tengchong City, Houqiao Town, on spider attached to underside of living leaf, 3 Nov. 2024, Q.F. Huang, H297 (holotype HKAS 148564, culture ex-type KUNCC25-19468).

Additional materials examined: China, Yunnan Province, Tengchong City, Houqiao Town, on a spider attached to underside of living leaf, 3 Nov. 2024, Q.F. Huang, H297b (HKAS 148562).

Notes: Chlorocillium houqiaoense, together with Ch. gibellulae and Ch. mauryae, forms a distinct clade remote from other members of this genus. Chlorocillium houqiaoense can be separated from Ch. gibellulae by its association with spiders, smaller phialides, and conidia, whereas the latter is a hyperparasite on synnemata of Gibellula sp. and has slenderer phialides and larger conidia (Table 2). Chlorocillium houqiaoense can be distinguished from Ch. mauryae by 12/1043 bp (1.1 %) differences in rpb2 sequences. Moreover, the PHI test (Fig. 4C3) shows that there is no significant recombination between species in this clade (Φw = 1.0), suggesting that they are different species.

Chlorocillium menghaiense D.P. Wei & T.C. Wen, sp. nov. MB 862661. Fig. 16.

Etymology: Named after the collection site of the type specimen, Menghai County in Xishuangbanna state of Yunnan Province.

Description: Hyperparasitic on Ophiocordyceps, Cordyceps species and parasitic on spiders. Asexual morph: Synnemata white to pale yellow-green, cottony to powdery, cylindrical, densely aggregated on fertile part of O. nutans, comprising hyaline, smooth-, thick-walled, parallelly arranged hyphae. Conidiophores developing from superficial hyphae of synnemata, micronematous, branched, septate, bearing solitary, verticillate to cluster of phialides. Phialides 8–18 ( $\overline{x} = 14$ , n = 50) μm long, base 1.3–2.8 ( $\overline{x} = 2.0$ , n = 35) μm wide, tip 0.6–1.1 ( $\overline{x} = 0.8$ , n = 40) μm wide, monophialidic, flask-shaped, narrowing into a thin neck, hyaline, smooth-walled, aseptate, straight or curved. Conidia 3.1–4.7 × 1.2–2.2 ( $\overline{x} = 3.8 \times 1.8$ , n = 40) μm, hyaline, ovoid with scar at one end, aseptate, smooth-walled, gathering in short chains. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 15 mm diam. after incubation for 60 d at 25 °C, white (1A1), floccose, circular, raised, mycelia moderately dense, edge entire, reverse coffee (6D3).

Typus: China, Yunnan Province, Menghai County, Gelanghe village, on fertile part of Ophiocordyceps nutans on leaf litter, 12 Sep. 2023, D.P. Wei, GLH43 (HKAS 135597).

Materials examined: China, Yunnan Province, Menghai County, Gelanghe village, on fertile part of Ophiocordyceps nutans on ground, 12 Sep. 2023, D.P. Wei, GLH44-1 (HKAS 135598), ibid., GLH52 (HKAS 135596); Pu’er City, Simao District, Xinfangshuiku Park, on stroma of O. nutans, 27 Oct. 2023, D.P. Wei, DSSZ51 (HKAS 135593, living culture KUNCC24-17916); Taiyanghe Park, on stroma of Cordyceps militaris, 23 Oct. 2023, D.P. Wei, TYHGY04 (HKAS 135600, living culture KUNCC24-17915); Xinping County, Ailaoshan national reserve, on stromata of Ophiocordyceps shimenxiaensis, 20 Oct. 2023, D.P. Wei, SMX07 (HKAS 148570); Tengchong City, Houqiao Town, on Ophiocordyceps sp. in leaf litter, 3 Nov. 2024, G.Y. Wang, DT10 (HKAS 148560, living culture KUNCC25-19469); Guizhou Province, Guiyang City, Huaxi District, Tongxin village, on fertile part of O. nutans, 15 Jun. 2018, D.P. Wei, TXC17FC (HKAS 135611).

Notes: Eight specimens occurring on the stromata of Ophiocordyceps and Cordyceps species were collected from Guizhou and Yunnan Provinces. These specimens form a clade sister to Chlorocillium neolepidopterorum (Fig. 4). However, our specimens differ from the type of Ch. neolepidopterorum in that they live as hyperparasitic fungi on the stromata of entomopathogenic fungi, whereas Ch. neolepidopterorum is a pathogen of lepidopteran larvae. In addition, our specimens produce ovoid conidia with a scar at one end and larger conidia (3.1–4.7 × 1.2–2.2 μm), whereas Ch. neolepidopterorum produces fusiform and smaller conidia (2.3–3.1 × 1.0–1.4). The results of the PHI test (Fig. 4C2) show no significant recombination (Φw = 1.0) between our specimens and Ch. neolepidopterorum, suggesting they are distinct species.

Chlorocillium neolepidopterorum W.H. Chen et al., Mycosphere 16(1): 2897. 2025. MB 903356. Fig. 17.

Replaced synonym: Metarhizium lepidopterorum W.H. Chen et al., Phytotaxa 419(2): 192. 2019.

Description: Hyperparasitic on Ophiocordyceps sp. Insect host 1.5 cm long, 0.5 cm wide, immersed into soil. Stroma of Ophiocordyceps sp. 4.5 cm long, stipitate. Stipe 3 cm long, 0.2 cm wide, off white, fleshy, glabrous, straight. Fertile part 0.4 cm in diam., terminal, globose, black brown, with black ostioles projecting on the surface. Asexual morph: Synnemata white to pale green, powdery, cylindrical, tapering toward the apex, densely aggregated on fertile part of Ophiocordyceps sp., comprising hyaline, smooth-walled, septate, parallelly arranged hyphae 1.4–2.6 ( $\overline{x} = 2.1$ , n = 25). Conidiophores micronematous, developing from superficial hyphae of synnemata, branched, septate, bearing solitary, verticillate to cluster of phialides. Phialides 9.3–24.0 ( $\overline{x} = 15$ , n = 35) μm long, base 1.8–3.5 ( $\overline{x} = 2.5$ , n = 50) μm wide, tip 0.7–1.6 ( $\overline{x} = 1.1$ , n = 50) μm wide, monophialidic, flask-shaped, narrowing into a thin neck, hyaline, smooth-walled, aseptate, straight or curved, tapering toward the apex. Conidia 3.3–6.0 × 1.8–2.7 ( $\overline{x} = 4.6 \times 2.3$ , n = 60) μm, hyaline, fusiform, aseptate, smooth-walled, gathering in short chains. Sexual morph: Undetermined.

Typus: China, Guizhou Province, Qiandongnan Miao and Dong Autonomous Prefecture, Rongjiang County, on a lepidopteran pupa, 1 Oct. 2018, W. Chen (holotype GZAC DL1022, culture ex-type GZAC DL10221).

Material examined: China, Yunnan Province, Tengchong City, Houqiao Town, on stroma of Ophiocordyceps sp., 3 Nov. 2024, N.Y. Liu, LWF2653 (HKAS 148561).

Notes: Metarhizium lepidopterorum was originally described by Chen et al. (2019) from a lepidopteran pupa in Rongjiang County, Guizhou Province. Subsequently, Chen et al. (2025) transferred this species to the genus Chlorocillium, renaming it Ch. neolepidopterorum. In the present study, specimen HKAS 148561 forms a clade with Ch. neolepidopterorum, which is sister to Ch. menghaiense. Morphologically, our specimen is similar to Ch. neolepidopterorum in producing white to pale green, cylindrical, powdery synnemata, flask-shaped phialides with thin necks, and fusiform conidia. Herein, we propose our specimen as a new host record for Ophiocordyceps sp.

Chlorocillium puerense D.P. Wei, Z. L. Liu & T.C. Wen, sp. nov. MB 859509. Fig. 18.

Etymology: Refers to Pu’er City in Yunnan Province, where the type specimen was collected.

Description: Hyperparasitic on hevansia-like taxon. Asexual morph: Colonies on host are white, fluffy. Conidiophores directly emerging from synnemata of hevansia-like taxon, complex, branched, bearing solitary or whorls of phialides. Phialides 13–47 ( $\overline{x} = 29$ , n = 30) μm long, base 0.8–1.4 ( $\overline{x} = 1.1$ , n = 30) μm wide, tip 0.3–0.7 ( $\overline{x} = 0.5$ , n = 30) μm wide, monophialidic, subulate, smooth-walled, gradually tapering toward apex, commonly producing small secondary branches or denticles, giving an appearance of zigzag on upper part of phialides. Conidia 2.0–3.0 × 0.8–1.3 ( $\overline{x} = 2.6 \times 1.0$ , n = 25) μm, hyaline, oblong, straight, aseptate, smooth-walled. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 10 mm diam. after incubation for 60 d at 25 °C, white (1A1), velvety, circular, mycelia dense, slightly raised, surface smooth, edge entire, reverse reddish golden (6C7).

Typus: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on hevansia-like fungus which infects spider attached to living leaves of monocotyledonous plant, 27 Oct. 2023, Z.L. Liu, MZH20231027198L (holotype HKAS 135606, culture ex-type KUNCC24-17917).

Notes: The phylogenetic analyses revealed that Ch. puerense is sister to Ch. montefioreae with strong statistical support (SH-aLRT = 99 %, PP = 1.00, UFB = 100 %; Fig. 4). Chlorocillium montefioreae was introduced by Tan & Shivas (2024) from a dead arachnid in Australia. However, the authors did not provide the morphological description for this species, making it impossible to compare their morphologies. Nevertheless, the nucleotide comparison between Ch. puerense and Ch. montefioreae revealed that there are six bp differences (including three gaps) within the 554 bp ITS, eight bp differences within the 785 bp tef-1α, and 19 bp differences within the 902 bp rpb2 region, suggesting these two species are different. Hence, we introduce Ch. puerense as a new species of Chlorocillium.

Chlorocillium sanduense D.P. Wei & T.C. Wen, sp. nov. MB 859511. Fig. 19.

Etymology: Named after Sandu County in Guizhou Province where the type specimen was collected.

Description: Hyperparasitic on Ascopolyporus sanduensis (Cordycipitaceae). Asexual morph: Colony entirely covering the stromata of Ascopolyporus sp., green, powdery, comprising hyaline, septate, branched hyphae. Conidiophores micronematous, irregularly branched, septate, bearing 1–3 phialides per nodes, complex. Phialides 14–45 ( $\overline{x} = 24$ , n = 35) μm long, base 1.1–1.8 ( $\overline{x} = 1.4$ , n = 40) μm wide, tip 0.4–0.8 ( $\overline{x} = 0.6$ , n = 35) μm wide, monophialidic, subulate, gradually tapering toward the apex, hyaline, smooth-walled, aseptate, typically curved. Conidia 2.5–3.7 × 1.0–1.5 ( $\overline{x} = 3.1 \times 1.3$ , n = 40) μm, hyaline, fusiform with scar at one end, aseptate, smooth-walled, gathering in chains. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 20 mm diam. after incubation for 60 d at 25 °C, white (1A1), circular, raised, mycelia dense, surface lumpy, radially striated with lobate edge, reverse reddish golden (6C7).

Typus: China, Guizhou Province, Sandu Shui Autonomous County, Yaorenshan Park, on stromata of Ascopolyporus sanduensis attached to a living stem of Pueraria sp., 24 Aug. 2023, D.P. Wei, SD0806 (holotype HKAS 135608, culture ex-type KUNCC24-17920).

Additional materials examined: China, Guizhou Province, Sandu Shui Autonomous County, Yaorenshan Park, on stromata of Ascopolyporus sanduensis attached to a living stem of Pueraria sp., 24 Aug. 2023, D.P. Wei, SD0805 (HKAS 135607, living culture KUNCC24-17919).

Notes: Chlorocillium sanduense is characterized by its yellow green colonies on the stromata of Ascopolyporus sanduensis and the absence of synnemata. Phylogenetically, this species was accommodated in a distinct clade sister to the Ch. guizhouense complex.

Chlorocillium taiyangheense D.P. Wei & T.C. Wen, sp. nov. MB 859513. Fig. 20.

Etymology: Named after the collection site of the type specimen, Taiyanghe Park in Pu’er City, Yunnan Province.

Description: Parasitic on a spider and Ophiocordyceps cf. oxycephala. Asexual morph: Synnemata white, fluffy to powdery, comprising hyaline, septate, parallel-arranged hyphae. Conidiophores micronematous, branched, septate, bearing solitary to verticillate phialides. Phialides 7.6–18.0 ( $\overline{x} = 11.0$ , n = 50) μm long, base 1.0–2.0 ( $\overline{x} = 1.4$ , n = 50) μm wide, tip 0.4–0.8 ( $\overline{x} = 0.6$ , n = 50) μm wide, monophialidic, flask-shaped, gradually tapering toward the apex, hyaline, smooth-walled, aseptate, typically curved. Conidia 2.4–3.8 × 0.8–1.3 ( $\overline{x} = 3.0 \times 1.1$ , n = 50) μm, hyaline, fusiform, aseptate, smooth-walled, gathering in chains. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 15 mm diam. after incubation for 60 d at 25 °C, white (1A1), circular, velvety, mycelia immersed into agar, margin entire, reverse reddish ochre (7D8).

Typus: China, Yunnan Province, Pu’er City, Simao District, Taiyanghe Park, on spider attached to living leaf of dicotyledonous plant, 23 Oct. 2023, D.P. Wei, TYHGY19 (holotype HKAS 135602, culture ex-type KUNCC24-17914).

Additional materials examined: China, Yunnan Province, Tengchong City, Mangbang Town, Zhenshan Village, Ganlusi Reservoir, on stromata of Ophiocordyceps cf. oxycephala, 2 Nov. 2024, N.Y. Liu, LWF3329 (HKAS 148568).

Notes: Chlorocillium taiyangheense is a sister to species of Ch. globiceps and shows a close relationship to Ch. griseum, Chlorocillium aschersoniae, and Ch. guizhouense (Fig. 4). The PHI test revealed that there is no significant recombination (Φw = 0.2108, Fig. 4C1) between Ch. taiyangheense and its closely related species. Interestingly, species in this group are dissimilar in macromorphology and associated hosts but are genetically very close (Zare & Gams 2016, Chen et al. 2025a). Chlorocillium globiceps is specific to Ophiocordyceps cf. globiceps and exhibits heavy sporulation on natural hosts; Ch. griseum is a generalist that produces greenish ochraceous colonies on spiders and various other hosts (Zare & Gams 2016). Chlorocillium guizhouense produces yellow-green, powdery colonies and cylindrical, gregarious synnemata, and is specific to spider hosts attached to tree bark. Chlorocillium taiyangheense produces white, velvety, fasciculate synnemata on spider hosts. It is worth noting that the spider hosts of Ch. guizhouense and Ch. taiyangheense are different species, based on evidence from their death locations: the host of Ch. guizhouense dies on tree bark, while the latter dies on the lower side of living leaves. This finding suggests that integrating ecological information can sometimes be useful in the taxonomic classification of species in Chlorocillium species.

Chlorocillium tengchongense D.P. Wei, N.Y. Liu & T.C. Wen, sp. nov. MB 859514. Fig. 21.

Etymology: Named after the collection site of the type specimen, Tengchong City of Yunnan Province.

Description: Parasitic on a lepidopteran larva. Asexual morph: Colonies on host are white, fluffy, covering the hairs of lepidopteran larvae. Conidiophores developing from hyphae of colonies, micronematous, mononematous, hyaline, branched, bearing solitary phialides. Phialides 25–79 ( $\overline{x} = 53$ , n = 40) μm long, base 2.6–4.8 ( $\overline{x} = 3.4$ , n = 40) μm wide, tip 1.6–2.6 ( $\overline{x} = 2.0$ , n = 40) μm wide, monophialidic, smooth-walled, straight to flexuous, subulate, gradually tapering toward apex. Conidia 3.8–10.5 × 1.7–4.0 ( $\overline{x} = 6.2 \times 2.6$ , n = 50) μm, hyaline, ovoid with one pointed end and a round end, unicellular, smooth-walled. Sexual morph: Undetermined.

Typus: China, Yunnan Province, Tengchong City, Zhonghe Town, Xibian village, on lepidopteran larvae, 1 Nov. 2024, N.Y. Liu, LWF3335 (holotype HKAS 148567).

Additional materials examined: China, Yunnan Province, Tengchong City, Zhonghe Town, Xibian village, on lepidopteran larvae, 1 Nov. 2024, N.Y. Liu, LWF3335b (HKAS 148566).

Notes: The multigene phylogeny showed that Chlorocillium tengchongense is sister to Ch. neorongjiangense, forming a clade that is closely related to Ch. cordycipiticola (Fig. 4). Species in this clade share similarities in that all three species have lecanicillium-like conidiophores and ovoid conidia with one round end and one-pointed end. However, they are different in their substrates and macro-morphology. Chlorocillium tengchongense directly forms white colonies on the hair of insect larvae, Ch. neorongjiangense produces robust synnemata from insect pupae, and Ch. cordycipiticola colonizes the stromata of Cordyceps cf. neopruinosa. A comparison of rpb2 sequences shows that Ch. tengchongense differs from Ch. neorongjiangense and Ch. cordycipiticola by 17/1040 bp (1.6 %) and 14/1031 bp (1.3 %), respectively. In addition, the PHI test showed that there is no significant recombination (Φw = 1.0, Fig. 4C4) between Ch. tengchongense, Ch. neorongjiangense and Ch. cordycipiticola.

Fiorinimazzantia Y.P. Tan et al., Index of Australian Fungi 22: 12. 2023. MB 901434

Diagnosis: Differs from Albacillium, Husseyia, Rousseaua and Speluncomyces in forming transparent, sparse colonies on stroma of host fungi.

Type species: Fiorinimazzantia elisabettae Y.P. Tan et al., Index of Australian Fungi 22: 12. 2023.

Description: Colony white, fluffy, sparse, covering the host. Phialides monophialidic, cylindrical, gradually attenuated toward the apex, slender, hyaline, smooth-walled, aseptate, straight or flexuous, arising from procumbent mycelia. Conidia hyaline, ellipsoidal with round ends, straight or slightly curved, asymmetric, aseptate, smooth-walled, gathering slimy head.

Notes: Fiorinimazzantia is a genus of hyperparasites, comprising F. elisabettae and F. australiana, which were reported on hirsutella-like fungus and Aschersonia mackerrasiae, respectively (Tan & Shivas 2023a, b). These two species were identified solely based on molecular data. In this study, we have obtained a fresh collection of F. elisabettae from Yunnan, China and provided a detailed morphological description for the type species of the genus. This genus was initially placed in an uncertain position within Sordariomycetes (Tan & Shivas, 2023a, b) and it was transferred to Chlorocilliaceae in this study (Fig. 3).

Fiorinimazzantia elisabettae Y.P. Tan et al., Index of Australian Fungi 22: 12. 2023. MB 901435. Fig. 22.

Description: Hyperparasitic on Hevansia minuta. Asexual morph: Colonies white, fluffy, sparse, covering the stroma of host fungus. Phialides 20–36 ( $\overline{x} = 26$ , n = 30) μm long, base 0.8–1.6 ( $\overline{x} = 1.3$ , n = 30) μm wide, tip 0.4–0.7 ( $\overline{x} = 0.5$ , n = 30) μm wide, monophialidic, lanceolate, slender, hyaline, smooth-walled, aseptate, straight or flexuous, arising from procumbent mycelia. Conidia 2.3–3.6 × 0.6–1.4 ( $\overline{x} = 3.0 \times 0.8$ , n = 40) μm, hyaline, allantoid with round ends, straight or slightly curved, asymmetric, aseptate, smooth-walled, gathering in slimy head. Sexual morph: Undetermined.

Culture characteristics: colonies on PDA attaining 17 mm diam. after incubation for 60 d at 25 °C, white (1A1), velvety, nearly circular, convex, mycelia immersed to aerial with moderately dense, edge erose, reverse honey yellow (4D6).

Typus: Australia, Queensland, Tully, on an unidentified dead insect colonized by an unidentified hirsutella-like fungus, 29 Apr. 2021, T.S. Marney et al. (holotype BRIP 72660a).

Materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on Hevansia minuta attached to living leaves of dicotyledonous plant, 27 Oct. 2023, Z.L. Liu, MZH20231027194L (HKAS 135605, living culture KUNCC24-17921).

Notes: Tan & Shivas (2023a) introduced Fiorinimazzantia elisabettae, with a photograph depicting its colony morphology on PDA agar. The authors speculated that this species infected an unidentified hirsutella-like fungus. In our study, we obtained a new collection with a similar appearance to the one outlined in Tan & Shivas (2023a). Through observation, we found that the host of F. elisabettae is likely Hevansia minuta, as we observed many uninfected specimens of H. minuta in the natural environment. Morphological illustrations of F. elisabettae were not provided in Tan & Shivas (2023a), but the accession numbers of its ITS, LSU, rpb2, and tef-1α sequences are available in the protologue. Sequences of our collection groups with F. elisabettae in the phylogenetic tree with maximum support (Fig. 3), and nucleotide comparison shows that our collection is identical to F. elisabettae within 531 bp ITS and 830 bp LSU, but different by one bp in 908 bp tef-1α and six bp in 877 bp rpb2. The genetic similarity confirms that our collection and F. elisabettae are conspecific. Fiorinimazzantia elisabettae was originally found in Australia, and this is the first report of its discovery in China.

Albacillium M.M. Ding & L.J. Xu, Phytotaxa 650: 2. 2024. MB 850888.

Diagnosis: Differ from closely related genera in producing white, loose, velvety colony on PDA media.

Type species: Albacillium hingganense M.M. Ding & L.J. Xu, Phytotaxa 650: 2. 2024.

Description: Colonies on PDA white, velvety, margin entire, mycelia loose, reverse creamy yellow, Conidiophores micronematous, branched, arising from procumbent mycelia, bearing solitary phialides. Phialides monophialidic, cylindrical, smooth-walled, hyaline, unbranched. Conidia hyaline, crescent-shaped, asymmetrical, aseptate, gathering in slimy head.

Notes: The genus Albacillium is a monotypic genus introduced by Ding et al. (2024). Later, Lin et al. (2025) introduced the second species, A. fuzhouense from a coleopteran larva. In their analysis, Albacillium phylogenetically grouped with Chlorocillium and the authors treated this genus as a member of Clavicipitaceae. In this study, we transfer this genus to Chlorocilliaceae based on molecular analyses and morphological characteristics.

Albacillium hingganense M.M. Ding & L.J. Xu, Phytotaxa 650: 2. 2024. MB 850889. Fig. 23.

Description: Associated with sclerotium of Ophiocordyceps cf. unilateralis. Asexual morph: Conidiophores micronematous, branched, arising from procumbent hyphae. Phialides 10–28 ( $\overline{x} = 19$ , n = 20) μm long, base 0.7–1.6 ( $\overline{x} = 1.1$ , n = 20) μm wide, tip 0.4–0.9 ( $\overline{x} = 0.6$ , n = 20) μm wide, monophialidic, cylindrical, smooth-walled, hyaline, solitary, straight, unbranched. Conidia 1.6–2.4 × 0.8–1.2 ( $\overline{x} = 2.0 \times 1.0$ , n = 40) μm, hyaline, reniform, asymmetrical, aseptate, gathering in slimy head. Sexual morph: Undetermined.

Culture characteristics: Colonies on PDA attaining 65 mm diam. after incubation for 60 d at 25 °C, white (1A1), fluffy, circular, surface smooth, mycelia aerial, edge entire, reverse reddish ochre (7D8) at centre, golden yellow (5B7) at periphery.

Typus: China, Heilongjiang Province, Greater Hinggan Mountains, on leaf litter (Betula platyphylla, Larix gmelinii and Pinus sylvestris), 28 Sep. 2018, Z. Li et al. (holotype HMPHU 1246, culture ex-type CCTCC M 20232069).

Materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, isolated from sclerotium of Ophiocordyceps cf. unilateralis parasitic on ant attached to living leaf of dicotyledonous plant, 22 Oct. 2023, D.P. Wei, XFSK61 (HKAS 135603, living culture KUNCC24-17922); ibid., XFSKB61b (HKAS 148545); ibid., XFSK61c (HKAS 148544).

Notes: Phylogenetic analysis shows that our strains group with Albacillium hingganense with maximum statistical support (Fig. 3), suggesting they are conspecific. Morphologically, our strain resembles A. hingganense in the nearly cylindrical phialides and ellipsoidal to cymbiform conidia that are gathered in mucoid globose heads. Ding et al. (2024) reported Albacillium hingganense from forest litter in northeast China. This is the first report of A. hingganense from the sclerotium of Ophiocordyceps cf. unilateralis.

Husseyia Y.P. Tan et al., Index of Australian Fungi 22: 13. 2023. MB 901436.

Diagnosis: Differs from Albacillium, Fiorinimazzantia, Rousseaua, Speluncomyces in producing phialides that are branched at apical region.

Type species: Husseyia annamariae Y.P. Tan et al., Index of Australian Fungi 22: 13. 2023.

Description: Colonies white, floccose, covering synnemata and perithecia of host. Conidiophores micronematous, branched, usually bearing solitary phialide. Phialides monophialidic, subulate, gradually tapering toward the apex, hyaline, smooth-walled, aseptate, apically branched or not. Conidia hyaline, crescent, aseptate, curved, smooth-walled, gathering in slimy head.

Notes: Husseyia was proposed by Tan & Shivas (2023a) based on Husseyia annamariae, which was isolated from an unidentified hevansia-like taxon in Australia. In the same year, Tan & Shivas (2023b) added the second species to the genus, Husseyia queenslandica, which was found on an unidentified dead arthropod in Australia. The protologue provides external images of these two species, which show that they both produce white, fluffy colonies on their natural hosts. However, additional macro- and microscopic characteristics were not fully described. The identification of both species was made solely through phylogenetic analysis (Tan & Shivas 2023a, b).

Husseyia ramosissima D.P. Wei & T.C. Wen, sp. nov. MB 859515. Fig. 24.

Etymology: Referring to the apically branched phialides.

Description: Hyperparasitic on Akanthomyceps sp. Asexual morph: Colonies white, fluffy, covering synnemata and perithecia of host species. Conidiophores micronematous, branched, usually bearing solitary phialide. Phialides 13–35 ( $\overline{x} = 23$ , n = 30) μm long, base 0.7–1.6 ( $\overline{x} = 1.0$ , n = 30) μm wide, tip 0.3–0.6 ( $\overline{x} = 0.4$ , n = 35) μm wide, monophialidic, subulate, gradually tapering toward the apex, hyaline, smooth-walled, aseptate, apically branched or not. Conidia 2.2–3.4 × 0.6–1.2 ( $\overline{x} = 2.6 \times 0.8$ , n = 25) μm, hyaline, reniform, aseptate, curved, smooth-walled, gathering in slimy head. Sexual morph: Undetermined.

Typus: China, Guizhou Province, Qiannan Buyi and Miao Autonomous Prefecture, Sandu Shui Autonomous County, Yaorenshan Park, on Akanthomyces sp. parasitic to moth attached to living leaf of bamboo, 30 Sep. 2023, D.P. Wei, YRS1012 (holotype HKAS 135601).

Notes: Husseyia ramosissima is sister to H. annamariae with maximum statistical support (Fig. 3). Husseyia annamariae produces white colonies on its natural substrate, but its micro-morphological illustration was not given in the protologue, making it impossible to compare it with H. ramosissima. A nucleotide comparison between H. ramosissima and H. annamariae shows that there are eight bp differences (including two gaps) within 518 bp ITS and 8 bp differences within 851 bp tef-1α. It is worth noting that H. ramosissima colonizes Akanthomyces sp., which infects moths in China, while H. annamariae was isolated from an unidentified hevansia-like species that colonized a dead spider in Australia (Tan & Shivas 2023a).

Neoaraneomyces Wan H. Chen et al., MycoKeys 91: 58. 2022. MB 842644.

Diagnosis: Differs from Paraneoaraneomyces and Subuliphorum in forming white, velvety colonies on spider hosts with dense mycelia on PDA medium.

Notes: Neoaraneomyces was established by Zhang et al. (2023), with Neoaraneomyces araneicola as the type species. Later, Lin et al. (2025) introduced a second species, Neoaraneomyces wuyishanensis, to the genus. Both species are araneogenous fungi, but they exhibit significant morphological differences. Neoaraneomyces araneicola produces fusiform conidia that aggregate in chains, whereas N. wuyishanensis forms lunate conidia that adhere in a slimy head. Phylogenetic analyses show that Neoaraneomyces form a monophyletic clade with Paraneoaraneomyces and occupy a basal position within the family Clavicipitaceae, as reported by Zhang et al. (2023) and Lin et al. (2025). In the present study, Neoaraneomyces is sister to a clade containing Paraneoaraneomyces and Subuliphorum, forming a distinct clade at the basal position of the newly proposed family Chlorocilliaceae.

Paraneoaraneomyces Z. Yuan Zhang & Y.F. Han, MycoKeys 98: 120. 2023. MB 848089.

Diagnosis: Differs from Neoaraneomyces and Subuliphorum by dwelling in soil.

Notes: Paraneoaraneomyces is a monotypic genus, containing a single species, Paraneoaraneomyces sinensis, which was isolated from soil in the green belt (Zhang et al. 2023). Paraneoaraneomyces produces solitary phialides from aerial mycelia or hyphae, a feature not observed in closely related genera. Initially, this genus was assigned to Clavicipitaceae due to its close affinity with Speluncomyces. Our findings reveal that this genus grouped with other genera of the newly proposed family Chlorocilliaceae. Morphologically, P. sinensis produces the solitary, lanceolate phialides, along with the cymbiform to reniform conidia, which align well with the characteristics of Chlorocilliaceae. Therefore, we formally transfer Paraneoaraneomyces to the family Chlorocilliaceae.

Speluncomyces A.F. Leão et al., Fungal Diversity 134: 319. 2025. MB 850531.

Diagnosis: Differs from Albacillium and Rousseaua in the special habitat — leaf litter in a ferruginous cave.

Notes: The phylogenetic tree constructed by Dong et al. (2025) shows that Speluncomyces forms a distinct clade, sister to Pseudometarhizium, which has been synonymized with Chlorocillium. In this study, Speluncomyces is closely related to Albacillium and Rousseaua. Morphologically, these three genera exhibit similarities in the structure of phialides and conidia, supporting their placement in the same family.

Subuliphorum L.W. Hou et al., Stud. Mycol. 105: 50. 2023. MB 845792.

Diagnosis: Differs from Neoaraneomyces and Paraneoaraneomyces in producing conidiophores that are unbranched or basitonously branched, or repeatedly verticillate towards the apex.

Notes: Subuliphorum was established to accommodate a species of Acremonium s. lat., A. camptosporum, which is phylogenetically distantly related to Acremonium s. str. (Hou et al. 2023). Subuliphorum is placed within a clade that nested between Neoaraneomyces and Paraneoaraneomyces in the study by Dong et al. (2025), and the same result was obtained in our study. Therefore, we include these aforementioned genera in the family Chlorocilliaceae. In contrast to Neoaraneomyces and Paraneoaraneomyces, Subuliphorum camptosporum has been reported from a broader range of hosts and substrates, including aerial contaminants, insects, nematodes, soil, mites, and it also acts as an endophyte of Bursera simaruba (Gonzalez et al. 2016, Hou et al. 2023).

Rousseaua Y.P. Tan & Marney, Index of Australian Fungi 27: 17. 2023. MB 900690.

Diagnosis: Differs from Albacillium and Speluncomyces in that it is a pathogenic fungus infecting Sternorrhyncha (Hemiptera).

Notes: Rousseaua was established by Tan & Shivas (2023b) based on Rousseaua marietteae, the type species of the genus. The protologue did not provide detailed illustrations of R. marietteae. However, from the plate provided by the authors, this species forms white, flat colonies with entire margins on PDA media. Lunate conidia were also observed in the culture, but the conidiophore and phialides were not depicted in the protologue. In our phylogenetic analysis, Rousseaua, Albacillium, and Speluncomyces form a monophyletic clade sister to Husseyia. The morphological characteristics of Rousseaua are like those of its closely related genera, Albacillium and Speluncomyces (Ding et al. 2024, Lin et al. 2025). Based on both morpho-phylogenetic analysis, these three genera are supported as members of the family Chlorocilliaceae.

Clavicipitaceae Rogerson, Mycologia 62(5): 900. 1970. MB 82061.

Notes: Clavicipitaceae is a family within Hypocreales, comprising 56 genera (Hyde et al. 2024). Members of this family exhibit a broad host range, including animals, plants, and fungi, and occupy diverse ecological roles, such as saprobic, endophytic, symbiotic, parasitic, and hyperparasitic lifestyles (Kepler et al. 2012, Lee et al. 2021, Liu et al. 2021b, Yapa et al. 2024). The functional diversity of Clavicipitaceae underscores its evolutionary success and significance in natural ecosystems, with ongoing research uncovering new genera, ecological roles, and biotechnological applications. In this study, 48 genera of Clavicipitaceae were included in the phylogenetic analysis (Fig. 3). The phylogenetic tree showed that 40 genera form the core clade of this family. Six genera, along with three other genera of Sordariomycetes genera incertae sedis, form a basal lineage adjacent to the core clade of Clavicipitaceae. We therefore propose a new family, Chlorocilliaceae, to accommodate the genera within this lineage. Notably, Metarhiziopsis was resolved as a distinct clade, sister to Ophiocordycipitaceae. Polymicrospora was placed in a clade sister to a group consisting of Metarhiziopsis, Niessliaceae, Pseudoniessliaceae, and Ophiocordycipitaceae.

Metarhizium Sorokin, Veg. Parasitenk. Mensch Tieren 2: 268. 1879. MB 8912.

Notes: Metarhizium is a genus of Clavicipitaceae, comprising approximately 70 species (Hyde et al. 2024). This genus is typified by Metarhizium anisopliae, an economically important species in biocontrol (Mongkolsamrit et al. 2020). However, many taxa historically assigned to Metarhizium anisopliae were later confirmed to constitute a species complex. Mongkolsamrit et al. (2020) revisited this genus and shed light on its interspecific phylogenetic relationships through expanded taxon sampling. The asexual morph of Metarhizium is characterized by the production of cylindrical, green conidia that typically aggregate into columns (Mongkolsamrit et al. 2020, Kobmoo et al. 2024). In this study, we collected a specimen exhibiting typical Metarhizium morphology. Molecular analyses and morphological observations further identified it as Metarhizium puerense. Notably, the stroma of M. puerense was colonized by a fungus with yellow conidial mass, which was identified herein as Pleurocordyceps neoagarica.

Metarhizium puerense Hong Yu bis et al., Biodivers. Data J. 12(e129087): 4. 2024. MB 852903. Fig. 30.

Description: Stroma up to 11 cm long, cylindrical, slightly narrow toward the apex, erect, solitary, branched at base, stipitate. Fertile part up to 5.3 cm long, 1 cm wide at the broadest portion, paired, cylindrical, covered by green, powdery hymenium. Stipe up to 5.7 cm long, 0.9 cm wide, brown, fibrous, glabrous, flexuous, rhizoid. Conidiophores micronematous, septate, terminating in branches with one to two phialides per branch. Phialides 8.4–19.0 × 1.3–2.7 ( $\overline{x} = 12.0 \times 2.1$ , n = 20) μm, monophialidic, smooth-walled, cylindrical with semi-papillate, thickened apices. Conidia 4.4–8.8 × 0.8–2.0 ( $\overline{x} = 6.8 \times 1.3$ , n = 50) μm, cylindrical with round ends, hyaline, smooth-walled, aseptate, aggregated in chains that are arranged in parallel columns.

Typus: China, Yunnan Province, Pu’er City, Simao District, on larvae of Lepidoptera, 2 Aug. 2023, H. Yu, (holotype YHH MP2308031, culture ex-type YFCCMP 9458).

Materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on lepidopteran larva buried in soil, 24 Oct. 2023, D.P. Wei, MZH25 (HKAS 148550).

Notes: Two sequences representing HKAS 148550S and HKAS 148550G, were generated from the stipe and fertile part of the specimen, respectively. Multigene phylogenetic analysis (Fig. 3) revealed that HKAS 148550S and HKAS 148550G cluster with Metarhizium puerense to form a clade sister to M. candelabrum. Morphologically, M. candelabrum can be distinguished from our specimen by the absence of stroma, smaller phialides (6–7.5 × 2–2.5 μm vs 8.4–19 × 1.3–2.7 μm), and its association with leafhoppers (Mongkolsamrit et al. 2020). In contrast, our specimen shares key traits with M. puerense, including robust, erect stromata, a green, powdery hymenium, cylindrical conidia, and an association with lepidopteran larvae in the same habitat. Furthermore, the dimensions of phialides and conidia in M. puerense overlap with those of our specimen. Notably, Ma et al. (2024) successfully isolated M. puerense from natural samples. However, attempts to isolate this fungus from stromata colonized by Pleurocordyceps neoagarica were unsuccessful, suggesting that the hyperparasite may inhibit the development or viability of its host fungus.

Cordycipitaceae Kreisel ex G.H. Sung et al., Stud. Mycol. 57: 48. 2007. MB 81701.

Notes: Since the establishment of the Cordycipitaceae by Sung et al. (2007), the generic composition within this family has undergone fundamental taxonomic revisions based on multigene phylogenetic analyses (Kepler et al., 2017, Wang et al. 2020). There are 38 recognized genera within Cordycipitaceae, including Akanthomyces, Ascopolyporus, Beauveria, Blackwellomyces, Cordyceps, Engyodontium, Gibellula, Hevansia, Hyperdermium, Parengyodontium, Simplicillium (Kepler et al. 2017), Leptobacillium (Zare & Gams 2016), Samsoniella (Mongkolsamrit et al. 2018), Neobaryopsis (Flakus et al. 2019), Flavocillium (Wang et al. 2020), Liangia (Wang et al. 2020), Neotorrubiella (Thanakitpipattana et al. 2020), Gamszarea (Zhang et al. 2021), Neohyperdermium (Thanakitpipattana et al. 2022), Jenniferia (Mongkolsamrit et al. 2022), Parahevansia (Mongkolsamrit et al. 2022), Pseudolecanicillium (Alves et al. 2022), Niveomyces (Araújo et al. 2022), Polystromomyces (Mongkolsamrit et al. 2022), Bhushaniella (Mongkolsamrit et al. 2023), Gamszarella (Crous et al. 2023), Pseudoniveomyces (Kobmoo et al. 2023), Matutinistella (Custódio & Pereira 2025), Arachnidicola, Corniculantispora, Corpulentispora, Kanoksria, Zarea, Zouia, and Lecanicillium (Khonsanit et al. 2024), Dimorphspora, Neolecanicillium, Nuciformispora, Paracorniculantispora, Paracorniculantispora, and Zongqiomyces (Chen et al. 2025). Members of this family exhibit diverse morphological characteristics and are associated with a wide array of hosts across the fungal, plant, and animal kingdoms, including soil-borne and environmental fungi. Extensive surveys have focused on the discovery of new taxa from arthropod hosts, while the taxonomic studies of hyperparasites on the stromata, synnemata, or subicula of other Cordycipitaceae species remain poorly understood. This study opens a new perspective on the ecological niches of Cordycipitaceae species, introducing a novel genus based on a hyperparasitic species found on the synnemata of Akanthomyces species and a new species of Ascopolyporus.

Ascopolyporus Möller, Bot. Mitt. Trop. 9: 300. 1901. MB 371.

Notes: Ascopolyporus was established by Möller (1901) to encompass four species: Ascopolyporus moellerianus, A. polychrous, A. polyporoides, and A. villosus. Subsequently, this genus was expanded to include an additional 10 species (Hennings 1902, 1909, Bischoff et al. 2005, Thanakitpipattana et al. 2022, Yu et al. 2023). Ascopolyporus is an epiphytic genus distinguished by its pulvinate, flattened to convex, polypore-like, subglobose stromata, semi-immersed to immersed perithecia, cylindrical asci with a thickened apex, and filiform ascospores. To date, a total of 14 species has been described within this genus (Yu et al. 2023). Of these, seven species have been confirmed by molecular data, while the phylogenetic affiliations of the remaining species remain doubtful due to a lack of molecular data. In the current study, we provide a detailed description of a new species of Ascopolyporus based on its sexual morph.

Ascopolyporus sanduensis D.P. Wei & T.C. Wen, sp. nov. MB 859516. Fig. 25.

Etymology: Named after the collection site where the type specimen was collected, Sandu County, Guizhou Province of China.

Description: Parasitic on scale insects. Sexual morph: Stromata up to 1.6 cm diam., 1 cm in thickness, epibiotic, pulvinate to hemispherical, upper surface white, dotted with ostioles, lower surface orange. Perithecia 268–385 × 107–158 ( $\overline{x} = 316 \times 128$ , n = 20) μm, semi-immersed, crowded, ovoid, apically connected by loosely interlaced, thin-walled hyphae. Peridium 11–25 ( $\overline{x} = 17$ , n = 20) μm, composed of hyaline, thick-walled cells of textura angularis. The texture of stromata is composed of hyaline cells of textura oblita to textura epidermoidea. Asci 125–228 × 4.5–6.3 ( $\overline{x} = 188 \times 5.3$ , n = 20) μm, cylindrical with a thickened cap, slightly attenuated toward the base. Asci cap 1.4–3.2 × 0.8–2.3 ( $\overline{x} = 2.5 \times 1.6$ , n = 40) μm, hemispherical, hyaline. Ascospores 79–208 × 0.3–0.8 ( $\overline{x} = 142 \times 0.5$ , n = 20) μm, hyaline, filiform, smooth-walled, aseptate, whole. Asexual morph: Undetermined.

Typus: China, Guizhou Province, Qiannan Buyi and Miao Autonomous Prefecture, Sandu Shui Autonomous County, Yaorenshan Park, on scale insects attached to living stem of Pueraria sp., 24 Aug. 2023, D.P. Wei, SD0807 (holotype HKAS 148575)

Additional materials examined: China, Guizhou Province, Qiannan Buyi and Miao Autonomous Prefecture, Sandu Shui Autonomous County, Yaorenshan Park, on scale insects attached to living stem of Pueraria sp., 24 Aug. 2023, D.P. Wei, SD0808 (HKAS 148576).

Notes: Ascopolyporus sanduensis forms densely clustered colonies on living stems of Pueraria sp., with portions of the stromata observed to be colonized by Chlorocillium sanduense. The multi-gene phylogenetic analysis shows that Ascopolyporus sanduensis forms a sister clade to Ascopolyporus galloides with strong statistical support (SH-aLRT = 99 %, PP = 1, UFB = 100 %, Fig. 3). Ascopolyporus sanduensis bears similarities with A. galloides in its association with scale insects and in producing subglobose to pulvinate stromata that are orange on the lower surface and white at the perithecial layers, as well as in having cylindrical asci and filiform, intact ascospores. The perithecia of A. sanduensis are ovoid, whereas they are obclavate in A. galloides. Additionally, the pairwise differences between A. sanduensis and A. galloides are 7/832 bp (0.8 %) in LSU, 11/485 bp (2.2 %) in ITS, 11/726 bp (1.5 %) in tef-1α, and 22/819 bp (2.6 %) in rpb2 sequences, respectively, supporting its novelty within this genus.

Paralecanicillium D.P. Wei & T.C. Wen, gen. nov. MB 859517.

Etymology: Referring to the morphological similarity to Lecanicillium but being phylogenetically distinct.

Type species: Paralecanicillium yunnanense D.P. Wei & T.C. Wen

Description: Colonies on natural substrate inconspicuous, consisting of transparent mycelia. Asexual morph: Conidiophores erect, emerging from synnemata of fungal host, macronematous, mononematous, solitary, unbranched, with thick-walled, septate, bifurcate, swollen base immersed into synnemata of fungal hosts, bearing two to three whorls of phialides. Phialides monophialidic, hyaline, smooth-walled, flask-shaped, with inflated basal portion and abruptly narrowing neck. Conidia hyaline, unicellular, smooth-walled, falciform. Sexual morph: Unknown.

Notes: Paralecanicillium bears resemblance to Lecanicillium species in the verticillate conidiophores. This genus forms a sister clade to Pleurodesmospora and show closely relationship with Akanthomyces s. str. However, species of Pleurodesmospora are characterized by denticulate conidiogenous cells, which differ dramatically from those of Paralecanicillium species. Akanthomyces s. str. commonly targets moths and produces ampulliform crowded phialides, which are densely compacted into a hymenium along the synnemata (Khonsanit et al. 2024). Thus, Paralecanicillium can be separated from Akanthomyces s. str. using both morphological and phylogenetic analyses.

Paralecanicillium yunnanense D.P. Wei & T.C. Wen, sp. nov. MB 859518. Fig. 26.

Etymology: Named after the province where the type specimen was collected, Yunnan Province of China.

Description: Hyperparasitic on hevansia-like fungus. Asexual morph: Colonies on natural substrate, inconspicuous, consisting of transparent mycelia. Conidiophores 81–137 ( $\overline{x} = 116$ , n = 10) μm long, erect, laterally arising from the synnemata of fungal host, macronematous, mononematous, solitary, with an aerial part and a basal part immersed into synnemata of fungal hosts. The aerial part of conidiophores up to 66 μm long, 1.1–1.9 ( $\overline{x} = 1.6$ , n = 20) wide, hyaline, thin-walled, unbranched, sparsely septate, bearing two to three whorls of phialides with two to eight phialides at per node and with interval of whorls being 2.2–3.4 × 0.6–1.2 ( $\overline{x} = 2.6 \times 0.8$ , n = 25) μm. The basal part of conidiophores up to 70 μm long, 2.2–5.3 ( $\overline{x} = 4.2$ , n = 20) wide, thick-walled, septate, bifurcate or unbranched, swollen, flexuous. Phialides 7.7–12.0 ( $\overline{x} = 9.7$ , n = 35) μm long, 1.5–2.3 ( $\overline{x} = 1.8$ , n = 35) μm wide at the broadest position, monophialidic, hyaline, smooth-walled, flask-shaped, with inflated basal portion and abruptly narrowing neck 4.3–6.5 × 0.5–0.9 ( $\overline{x} = 5.3 \times 0.7$ , n = 20) μm. Conidia 2.2–3.4 × 0.6–1.2 ( $\overline{x} = 2.6 \times 0.8$ , n = 25) μm, hyaline, unicellular, smooth-walled, straight or curved, falciform with pointed ends.

Culture characteristics: Colonies on PDA attaining 28 mm diam. after incubation for 60 d at 25 °C, white (1A1), velvety, circular, raised at centre, flat at periphery, edge feather, reverse yolk yellow (4B8).

Typus: China, Yunnan Province, Menghai County, Nanhai Village, on hevansia-like fungus parasitic on moth attached to living dicotyledonous leaves, 8 Sep. 2023, D.P. Wei, NHC08 (holotype HKAS 148557, culture ex-type KUNCC25-19470).

Additional materials examined: China, Yunnan Province, Menghai County, Nanhai Village, on hevansia-like fungus parasitic on moth attached to living dicotyledonous leaves, 8 Sep. 2023, D.P. Wei, NHC04 (HKAS 148556); ibid., NHC22 (HKAS 148558).

Notes: The multigene phylogenetic analysis shows that Paralecanicillium is sister to Pleurodesmospora, forming a clade closely related to Akanthomyces, Arachnidicola, Kanoksria and Lecanicillium with strong statistical support (SH-aLRT = 94 %, PP = 1, UFB = 100 %, Fig. 3). However, Paralecanicillium can be distinguished from these genera by its hyperparasitic association with hevansia-like fungi and conidiophores that are bifurcate and immersed in the synnemata of host fungi. Additionally, the phialides, which are inflated at the basal portion and arranged in whorls, and the falciform conidia are diagnostic characteristics of Paralecanicillium, while these features are absent in its allied genera.

Polycephalomycetaceae Y.P. Xiao et al., Fungal Diversity 120: 23. 2023. MB 559469.

Notes: Polycephalomycetaceae was delineated from the Ophiocordycipitaceae to encompass Perennicordyceps, Pleurocordyceps, and Polycephalomyces (Xiao et al. 2023). Subsequently, Dingleyomyces and Paradingleyomyces have been introduced to this family based on their respective phylogenetic attributes (Johnston & Park 2023, Wang et al. 2024). Many species of Polycephalomycetaceae are hyperparasitic to Ophiocordyceps species, while some are parasites of arthropods (Wei et al. 2022, Xiao et al. 2023, 2024, Chen et al. 2024b, Liu et al. 2024). In this study, we introduce a new genus, Neodingleyomyces in Polycephalomycetaceae and incorporate Torrubiellomyces into this family. Additionally, Ophiocordyceps sessilis is transferred to Torrubiellomyces.

Neodingleyomyces D.P. Wei, N.Y. Liu & T.C. Wen, gen. nov. MB 859519.

Etymology: Referring to the morphological similarity to Dingleyomyces but distinct in phylogeny.

Type species: Neodingleyomyces tengchongensis D.P. Wei, N.Y. Liu & T.C. Wen

Description: Colonies entirely covering the insect body and synnemata of fungal hosts, white, velvety. Sporodochia forming from colonies on insect body, discoid, white, with yellow α-conidia mass on the top of disc, superficial, crowded, comprising compact α-phialides and setae-like hyphae. Conidiophores hyaline, septate, thin-walled, integrated. Setae-like hyphae hyaline, clavate, branched, aseptate, verrucose, slightly flexuous. α-Phialides hyaline, smooth-walled, aseptate, flask-shaped, becoming narrow toward the apex, with secondary branches. α-Conidia hyaline, smooth-walled, aseptate, ellipsoidal, abundant. β-Phialides arising from hair of insect and synnemata of fungal host, hirsutella-like, with cylindrical base and a thin neck, hyaline. β-Conidia hyaline, ellipsoidal, aseptate, smooth-walled, guttulate.

Notes: Neodingleyomyces is sister to Dingleyomyces, and supported by robust statistical support (Fig. 3). Distinctly, Neodingleyomyces is characterized by the production of discoid sporodochia and dimorphic phialides, which are not observed in Dingleyomyces. Notably, the presence of setae-like hyphae is a unique feature not reported in any other species within Polycephalomycetaceae, readily distinguishing Neodingleyomyces from other members of this family.

Neodingleyomyces tengchongensis D.P. Wei, N.Y. Liu & T.C. Wen, sp. nov. MB 859520. Fig. 27.

Etymology: Referring to the collection sites where the type specimen was found, Tengchong City, Yunnan Province of China.

Description: Hyperparasitic on Ophiocordyceps species. Asexual morph: Colonies entirely covering the insect body and synnemata of Ophiocordyceps sp., white, velvety. Sporodochia 245–563 × 217–498 ( $\overline{x} = 448 \times 387$ , n = 15) μm, developing from colonies on insect body, discoid, white edge, with yellow α-conidia mass on the top, superficial, crowded, comprising compact α-phialides and setae-like hyphae. Conidiophores hyaline, smooth-walled, branched, terminating in cluster of α-phialides. Setae-like hyphae 13–48 × 2–3 ( $\overline{x} = 26 \times 2.4$ , n = 20) μm, hyaline, clavate, branched, aseptate, verrucose, slightly flexuous. α-Phialides 8.4–17 ( $\overline{x} = 12$ , n = 30) μm long, 1.2–2.0 ( $\overline{x} = 1.7$ , n = 30) μm wide near the base, 0.4–0.8 ( $\overline{x} = 0.6$ , n = 30) μm wide near to the apex, hyaline, smooth-walled, aseptate, flask-shaped, becoming narrow toward the apex, laterally giving rise to cylindrical secondary branches 5.5–30 × 0.4–0.8 ( $\overline{x} = 15 \times 0.6$ , n = 20) μm. α-Conidia 2.2–3.4 × 0.6–1.2 ( $\overline{x} = 2.6 \times 0.8$ , n = 25) μm, hyaline, smooth-walled, aseptate, ellipsoidal, abundant. β-Phialides 18–34 ( $\overline{x} = 26$ , n = 20) μm long, emerging from hair of insect and synnemata of fungal host, hirsutella-like, cylindrical at base 2.0–3.0 ( $\overline{x} = 2.4$ , n = 20) μm wide, arbitrarily narrowing into a thin neck 0.6–1.2 ( $\overline{x} = 0.9$ , n = 20) μm wide, hyaline. β-Conidia 3.2–4.3 × 1.3–2.0 ( $\overline{x} = 3.8 \times 1.6$ , n = 20) μm, singly forming at tip of β-phialides, hyaline, ellipsoidal, aseptate, smooth-walled, with one to two globose guttulate. Sexual morph: undetermined.

Typus: China, Yunnan Province, Tengchong City, Houqiao Town, Houqiao village, on Ophiocordyceps sp. attached to fallen dicotyledonous leaves, 3 Nov. 2024, N.Y. Liu, LWF3319 (holotype HKAS 148565).

Additional materials examined: China, Yunnan Province, Tengchong City, Houqiao Town, Houqiao village, on Ophiocordyceps sp. attached to fallen dicotyledonous leaves, 3 Nov. 2024, N.Y. Liu, 3319b (HKAS 148563).

Notes: Neodingleyomyces tengchongensis shares morphological features with other members of Polycephalomycetaceae in producing dimorphic phialides. However, N. tengchongensis produces macro- and microconidia that are similar in shape but differ in size (α-conidia: 2.2–3.4 × 0.6–1.2 vs β-conidia: 3.2–4.3 × 1.3–2), whereas the dimorphic conidia of other species within Polycephalomycetaceae typically exhibit contrasting shapes (Xiao et al. 2023). Additionally, setae-like hyphae interlaced with conidiophores are observed for the first time in this species. These unique characteristics collectively indicate that N. tengchongensis represents a novel addition to this family.

Pleurocordyceps Y.J. Yao et al., J. Syst. Evol. 59(5): 1074. 2021. MB 570683.

Notes: Pleurocordyceps is a genus segregated from Polycephalomyces, typified by Pl. sinensis. The members of Pleurocordyceps predominantly comprises hyperparasites on the stomata of Ophiocordyceps with several exceptions such as Pl. lanceolata (Xiao et al. 2023), Pl. lianzhouensis (Wang et al. 2021), Pl. onorei which have been reported on lepidopteran larva (Wang et al. 2021), Pl. marginaliradians on Cossidae larvae (Wang et al. 2021), Pl. multisynnema on Paraisaria (Xiao et al. 2024), Pl. parvicapitata on Elaphomyces (Xiao et al. 2023), Pl. phaothaiensis on coleopteran larvae (Wang et al. 2021), and Pl. ramosopulvinata on Cicadida (Wang et al. 2021). Pleurocordyceps is commonly found in leaf litter or embedded within the soil. To date, 22 species have been classified within this genus (MycoBank, accessed on 25 June 2025), all of which have molecular data and confirmed via phylogenetic analyses (GenBank, accession date: 25 June 2025). The asexual morph of this genus is characterized by dimorphic phialides and conidia (Wei et al. 2022, Xiao et al. 2023). The sexual morph is distinguished by the formation of fertile cushions that are laterally or terminally attached to the stromata, cylindrical asci with thickened apices, and filiform ascospores that disarticulate into cylindrical secondary part-spores (Wang et al. 2024). In this study, we introduce one new species and document three new host records for Pleurocordyceps.

Pleurocordyceps heilongtanensis Y.B. Wang et al., Fungal Diversity 120: 61. 2023. MB 559476. Fig. 28.

Description: Hyperparasitic on Ophiocordyceps nutans and Ophiocordyceps sp. or parasitic on spider or co-occurring with Ophiocordyceps crinalis. Synnemata of Ophiocordyceps sp. growing from dead spider, cylindrical, pale brown at base, becoming white toward the apex, unbranched, scattered. Phialide 6.3–11 × 3.1–5.4 ( $\overline{x} = 8.4 \times 4.5$ , n = 25) μm, densely compacted into a layer covering synnemata, subglobose to broadly oblong, yellow brown, verrucose. Conidia 6–10 × 1.8–2.4 ( $\overline{x} = 7.8 \times 2.2$ , n = 20) μm, singly forming from apex of phialides, hyaline, obovate, aseptate. Asexual morph: Hyphomycetous. Synnemata stipitate, cylindrical, scattered, unbranched, apically terminate in single, globose, yellowish conidia masses. Stipe 0.3–1 cm long, cylindrical, white, becoming pale brown at base, glabrous. Conidiophores synnematous, hyaline, septate, branched, arranged in a parallel palisade-like layer. α-phialides monophialidic, hyaline, smooth-walled, slender lanceolate, caespitose, crowed, laterally producing one to two secondary branches 10–72 × 0.6–1.0 ( $\overline{x} = 29 \times 0.8$ , n = 25) μm. β-phialides 22–93 ( $\overline{x} = 51$ , n = 20) μm long, 1.9–2.5 ( $\overline{x} = 2.2$ , n = 20) μm wide at base, 0.6–1.3 ( $\overline{x} = 0.9$ , n = 20) μm, directly emerging from synnemata of Ophiocordyceps sp., hyaline, smooth-walled, hirsutella-like, elongated lageniform, with a cylindrical base and a thin neck. α-conidia 2.2–3.4 × 0.6–1.2 ( $\overline{x} = 2.6 \times 0.8$ , n = 25) μm, hyaline, obclavate, aseptate. β-conidia 3.2–4.8 × 1.4–1.7 ( $\overline{x} = 4 \times 1.5$ , n = 20) μm, fusiform, hyaline. Sexual morph: undetermined.

Culture characteristics: Colonies on PDA attaining 45 mm diam. after incubation for 30 d at 25 °C, white (1A1), circular, flat, mycelia dense, edge entire, with erect synnemata forming at periphery in annular arrangement and clustering at centre, reverse apricot (5B6).

Typus: China, Yunnan Province, Kunming City, Heilongtan Park, parasitic on Ophiocordyceps sp. associated with the larva of Lepidoptera in the leaf litter, 30 Sep. 2021, Y.B. Wang, Y.B. Wang 07 (holotype HKAS 122650, culture ex-type KUMCC 3008).

Materials examined: China, Yunnan Province, Kunming City, Yeyahu Park, on spider, 4 Jul. 2021, D.P. Wei, YYH03 (HKAS 148573, living culture KUNCC25-19472); on Ophiocordyceps sp. parasitic on spider, 4 Jul. 2021, D.P. Wei, YYH01 (HKAS 148574, living culture KUNCC25-19471); Pu’er City, Simao District, Xinfangshuiku Park, co-occurring with Ophiocordyceps crinalis, 27 Oct. 2023, D.P. Wei, DSSZ59 (HKAS 148549); Taiyanghe Park, on stroma of Ophiocordyceps nutans, 23 Oct. 2023, D.P. Wei, TYHGY18 (HKAS 148554).

Notes: Pleurocordyceps heilongtanensis has been introduced as a hyperparasite on Ophiocordyceps sp. associated with lepidopteran larvae in leaf litter in Kunming City, Yunnan Province (Xiao et al. 2023). In this study, we further demonstrate that this species exhibits a broader host range, parasitizing Ophiocordyceps nutans, an unidentified Ophiocordyceps sp., and co-occurring with Ophiocordyceps crinalis. The phialides and conidia of the host fungus Ophiocordyceps sp. were observed and illustrated in Fig. 28 (Q, U), supporting its taxonomic placement within Ophiocordyceps. Micromorphologically, our specimens align with the type material of P. heilongtanensis, characterized by clavate α-conidia, fusiform β-conidia, and conidial dimensions falling within the same size range.

Pleurocordyceps parvicapitata Y.P. Xiao et al., Fungal Diversity 120: 50. 2023. MB 559473. Fig. 29.

Description: Hyperparasitic on Ophiocordyceps cf. myrmecophila. Asexual morph: Colony white, velvety. Sporodochia on natural host up to 661 μm in diameter, circular or path-like, white at edge, with yellowish conidial masses covering the surface. Conidiophores developing from vegetative hyphae, hyaline, apically bearing cluster of α-phialides. α-phialides 10–14 ( $\overline{x} = 11$ , n = 20) μm long, 1.1–2.3 ( $\overline{x} = 1.5$ , n = 20) μm wide at base, 0.5–0.9 ( $\overline{x} = 0.7$ , n = 20) μm wide near the tip, monophialidic, hyaline, narrowly lageniform, gradually attenuated toward the apex, smooth-walled, crowed, palisade-like in arrangement, producing cylindrical, symmetrical secondary branches 5–11 × 0.5–1.4 ( $\overline{x} = 8.2 \times 0.8$ , n = 20) μm. β-phialides 7–20 ( $\overline{x} = 14$ , n = 20) μm long, 1.0–2.3 ( $\overline{x} = 1.8$ , n = 20) μm wide at base, 0.6–1.3 ( $\overline{x} = 0.9$ , n = 20) μm wide near the tip, directly forming from edge of sporodochia, hirsutella-like, lageniform, with a cylindrical base and a thin neck. α-conidia 1.6–2.7 ( $\overline{x} = 2.0$ , n = 50) μm, hyaline, globose, aseptate. β-conidia 3.0–5.0 × 1.2–2.2 ( $\overline{x} = 3.8 \times 1.5$ , n = 20) μm, fusiform, hyaline, aseptate, smooth-walled. Sexual morph: undetermined.

Typus: China, Yunnan Province, Xishuangbanna City, Dadugang County, on Elaphomyces sp. (Elaphomycetaceae, Eurotiales) buried in soil, 11 Jun. 2018, Y.P. Xiao (holotype MFLU 21-0270).

Material examined: China, Yunnan Province, Pu’er City, Simao District, Taiyanghe Park, on stroma of Ophiocordyceps cf. myrmecophila, 23 Oct. 2023, D.P. Wei, TYHGY08 (HKAS 148555).

Notes: The phylogenetic analysis (Fig. 5) revealed that the new collection (HKAS 148555) clusters with the type of Pleurocordyceps parvicapitata (MFLU 21-0271). Furthermore, pairwise comparison of sequences shows that our specimen HKAS 148555 is identical to the type of P. parvicapitata in the LSU, ITS, SSU, and rpb2 gene regions, indicating that they are conspecific. Morphologically, our specimen closely resembles P. parvicapitata in producing globose α-conidia and fusiform β-conidia (Xiao et al. 2023, Wang et al. 2024). The sexual morph of P. parvicapitata has been documented as a parasite of Elaphomyces sp., while its asexual morph occurs on the stromata of Perennicordyceps elaphomyceticola, which is itself a parasite of Elaphomyces sp. in Xishuangbanna, Yunnan Province (Xiao et al. 2023). Wang et al. (2024) further reported this species in association with Elaphomyces sp. and P. elaphomyceticola in Tengchong City, Yunnan Province. In this study, we report the occurrence of this species on Ophiocordyceps cf. myrmecophila, for the first time, in Pu’er City, Yunnan Province.

Pleurocordyceps neoagarica Yu Yang & Yuan Pin Xiao, Frontiers Microbiol. 14 (no. 1256967): 6. 2024. MB 900450. Fig. 30.

Description: Hyperparasitic on Metarhizium puerensis. Asexual morph: Colonies on natural hosts are white, velvety, nearly circular or path-like, with yellowish conidial masses covering the stromata of fungal host. Conidiophores micronematous, hyaline, apically branched into phialides, densely compact into hymenium. α-phialides 8.6–21.0 ( $\overline{x} = 13.0$ , n = 30) μm long, 0.6–1.5 ( $\overline{x} = 1.2$ , n = 30) μm wide at base, 0.5–1.0 ( $\overline{x} = 0.8$ , n = 30) μm wide at tip, monophialidic, hyaline, narrowly lageniform, with slightly inflated base and thin neck, smooth-walled, arranged in palisade-like, crowed, producing cylindrical, symmetrical secondary branches 9–18 × 0.7–1.3 ( $\overline{x} = 18 \times 0.9$ , n = 30) μm. β-phialides 19–43 × 1.1–2.6 ( $\overline{x} = 29 \times 1.8$ , n = 20) μm, directly form from vegetative hyphae surrounding the hymenium, hirsutella-like, subulate, gradually attenuated toward the apices. α-conidia 1.1–2.3 × 0.7–1.1 ( $\overline{x} = 1.6 \times 0.9$ , n = 50) μm, hyaline, globose to broadly ovoid, aseptate, smooth-walled. β-conidia 3.0–5.5 × 1.3–2.3 ( $\overline{x} = 4.4 \times 1.9$ , n = 30) μm, fusiform, hyaline, aseptate, smooth-walled. Sexual morph: undetermined.

Culture characteristics: Colonies on PDA attaining 45 mm diam. after incubation for 30 d at 25 °C, cadmium orange (5A3) at the centre, white (1A1) at the periphery, fluffy, circular, with entire edges, producing yellow conidial masses in the central zone, forming concentric annuli on the surface, and reverse oxide yellow (5C7).

Typus: China, Guizhou Province, Qiannan Buyi and Miao Autonomous Prefecture, Sandu Shui Autonomous County, parasitic on Ophiocordyceps neogryllotalpae in the soil, 10 Apr. 2022, Y. Yang, SD10H (holotype GZLG 23-103, culture ex-type GZCC 22-2043).

Materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stroma of Metarhizium puerense, 24 Oct. 2023, D.P. Wei, MZH25Y (HKAS 148550, living culture KUNCC25-19473).

Notes: Our strain clustered into a monophyletic clade with Pleurocordyceps neoagarica (Fig. 5). Nucleotide sequence comparisons revealed that our strain shares nearly identical LSU, ITS, and SSU regions with P. neoagarica, strongly supporting their conspecificity. Pleurocordyceps neoagarica was first reported as a hyperparasite infecting Ophiocordyceps neogryllotalpae in Guizhou Province, China (Yang et al. 2024). Its original description, based on axenic culture studies, documented only α-conidia in the protologue. In this study, we provide the first documentation of P. neoagarica as a hyperparasite on Metarhizium puerense in Yunnan Province, China, and describe its dimorphic phialides and conidia observed in natural specimens.

Pleurocordyceps simaoensis D.P. Wei & T.C. Wen, sp. nov. MB 859521. Fig. 31.

Etymology: Referring to the collection sites where the type specimen was found, Simao District, Pu’er City, Yunnan Province of China.

Description: Hyperparasitic on Ophiocordyceps spp. Asexual morph: Sporodochia up to 417 μm in diam., developing from colonies on fertile part of O. nutans, orange, dot-like, superficial, crowded, comprising compact α-phialides. Synnemata 1.2 mm long, 184 μm wide at base, 37 μm wide at tip, growing on stipe of O. nutans and O. cf. sphecocephala, white, erect, cylindrical, attenuated toward the apex. Conidiophores micronematous, hyaline, smooth-walled, apically bearing cluster of α-phialides in palisade arrangement. α-Phialides 6.8–18.0 ( $\overline{x} = 11.0$ , n = 30) μm long, 1.0–3.0 ( $\overline{x} = 1.8$ , n = 30) μm wide near the base, 0.5–1.0 ( $\overline{x} = 0.8$ , n = 20) μm wide near to the apex, hyaline, thin-walled, aseptate, subulate, becoming narrow toward the apex, laterally giving raise to cylindrical, symmetrical secondary branches 6.5–19.0 × 0.7–1.8 ( $\overline{x} = 11.0 \times 1.2$ , n = 20) μm. α-Conidia 1.9–3.0 × 1.3–1.9 ( $\overline{x} = 2.4 \times 1.6$ , n = 25) μm, hyaline, smooth-walled, aseptate, broadly fusiform, abundant. β-Phialides 8–14 ( $\overline{x} = 10$ , n = 30) μm long, laterally forming from synnemata, hirsutella-like, flask-shaped, cylindrical at base 2–3 ( $\overline{x} = 2.5$ , n = 20) μm wide, arbitrarily narrowing into a thin neck 0.8–1.2 ( $\overline{x} = 1.0$ , n = 20) μm wide, hyaline. β-Conidia 3.2–4.8 × 1.2–2.2 ( $\overline{x} = 3.8 \times 1.6$ , n = 25) μm, hyaline, fusiform, aseptate, smooth-walled. Sexual morph: undetermined.

Typus: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on stroma of Ophiocordyceps nutans, 22 Oct. 2023, D.P. Wei, XFSK22 (holotype HKAS 148553).

Additional materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park, on Ophiocordyceps cf. sphecocephala, 22 Oct. 2023, D.P. Wei, XFSK33 (HKAS 148546).

Notes: the sequences HKAS 148553F1 and HKAS 148553F2 were respectively generated from white synnemata and yellow pulvinate sporodochia that grew on the stroma of Ophiocordyceps nutans. The sequence HKAS 148546 was derived from white synnemata on the stroma of Ophiocordyceps cf. sphecocephala. Phylogenetic analyses revealed that these three sequences form a distinct clade, sister to Pleurocordyceps sanduensis with robust statistical support (SH-aLRT = 88 %, PP = 0.99, UFB = 97 %; Fig. 5). Morphologically, our specimens differ from P. sanduensis in possessing broadly fusiform α-conidia and smaller β-phialides (8–14 × 2–3 μm vs 19–33.4 × 0.9–1.8 μm). In contrast, P. sanduensis is characterized by globose α-conidia and occurs as a hyperparasite of Ophiocordyceps neogryllotalpae, which is associated with larvae of Gryllotalpa spp. in Guizhou Province, China (Xiao et al. 2024).

Paradingleyomyces Y. Wang tris & T.C. Wen, MycoKeys 110: 193. 2024. MB 901540.

Notes: Paradingleyomyces is a monotypic genus, typified with Pa. lepidopterorum hyperparasitic on Ophiocordyceps cf. cochlidiicola (Wang et al. 2024). The type species was introduced based on its sexual morph. Herein we provide a detailed description for its asexual morph and identify its host fungus as Ophiocordyceps shimenxiaensis.

Paradingleyomyces lepidopterorum Y. Wang tris & T.C. Wen, MycoKeys 110: 193. 2024. MB 901541. Fig. 32.

Description: Hyperparasitic on Ophiocordyceps shimenxiaensis. Asexual morph: Subiculum white, cottony, covering stromata of O. shimenxiaensis. Perithecia growing from subiculum and perithecia of fungal host, creamy yellow, ovoid, superficial. Sporodochia up to 291 μm in diameter, 152 μm thickness, growing on perithecia of fungal host, white, pulvinate, velvety. Conidiophores micronematous, hyaline, laterally or terminally bearing solitary phialide, densely compact into a hymenium. Phialides 11–29 ( $\overline{x} = 18$ , n = 20) μm long, hirsutella-like, hyaline, smooth-walled, cylindrical at base 0.8–2.8 ( $\overline{x} = 1.7$ , n = 20) μm, arbitrarily narrowing into thin neck 0.6–1.2 ( $\overline{x} = 0.8$ , n = 20) μm. Conidia 3.0–6.2 × 0.9–1.5 ( $\overline{x} = 4.5 \times 1.3$ , n = 20) μm, hyaline, ovoid, ellipsoidal, aseptate. Sexual morph: see Wang et al. (2024).

Typus: China, Yunnan Province, Tengchong County, Houqiao Town, parasitic on the stromata of Ophiocordyceps cf. cochlidiicola, 5 Nov. 2022, Y. Wang, GYY543H (holotype HKAS 131926).

Materials examined: China, Yunnan Province, Pu’er City, Simao District, Xinfangshuiku Park,, on stromata of Ophiocordyceps shimenxiaensis, 20 Oct. 2023, D.P. Wei, MZH24h (HKAS 148547).

Notes: The sexual morph of Paradingleyomyces lepidopterorum has been documented by Wang et al. (2024), wherein the authors identified its host fungus as Ophiocordyceps cf. cochlidiicola based on morphological observations. We have collected one specimen similar to P. lepidopterorum in producing superficial, pale yellow perithecia on white subiculum and in living as a hyperparasite on Ophiocordyceps shimenxiaensis. The multilocus phylogenetic analysis (Fig. 5) shows that our new collection clustered with P. lepidopterorum, supporting conspecificity. Additionally, the comparison of nucleotide sequences shows that our specimen is identical to P. lepidopterorum in the LSU, ITS, tef-1α, and rpb1 regions. This is the first time to describe its asexual morph co-occurring with the sexual morphs from a natural specimen. The asexual morph is characterized by the formation of white, velvety sporodochia on perithecia and stromata of the host fungus.

Torrubiellomyces J.P.M. Araújo & de Bekker, Persoonia 49: 188. 2022. MB 844048.

Notes: Torrubiellomyces is a monotypic genus established by Araújo et al. (2022) to accommodate Torrubiellomyces zombiae, a mycoparasite of Ophiocordyceps camponoti-floridani. The genus is morphologically defined by dark brown to black, superficial perithecia, cylindrical asci, filiform multiseptate ascospores, and cuboid secondary ascospores. Notably, T. zombiae produces viscous conidial masses in culture, a trait shared with Pleurocordyceps species. Prior to the erection of Polycephalomycetaceae, Torrubiellomyces was accommodated in Ophiocordycipitaceae. Phylogenetic analyses by Araújo et al. (2022) revealed that Torrubiellomyces clusters with Polycephalomyces, Perennicordyceps, and Pleurocordyceps. These genera were subsequently transferred to Polycephalomycetaceae by Xiao et al. (2023). Our phylogenetic analysis generated results congruent with those of Araújo et al. (2022). The consistent grouping of Torrubiellomyces within Polycephalomycetaceae supports its formal taxonomic transfer to this family, based on combined morphological characteristics and phylogenetic evidence.

Torrubiellomyces sessilis (Kaitsu et al.) D.P. Wei, T.C. Wen, comb. nov. MB 859522.

Basionym: Ophiocordyceps sessilis Kaitsu et al., Mycol. Prog. 12(4): 759. 2013.

Description: See Kaitsu et al. (2013).

Typus: Japan, Fukushima Prefecture, on Camponotus obscuripes (Hymenoptera: Formicidae) clinging to twigs, 7 Oct. 2007, Y. Kaitsu (holotype CBM FA-39236, culture ex-type IFM-61904).

Notes: Ophiocordyceps sessilis was originally described as an entomopathogenic taxon parasitizing Camponotus obscuripes (Kaitsu et al. 2013). This species shares similarities with Torrubiellomyces zombiae, including the production of superficial dark brown perithecia, cylindrical asci, and filiform ascospores that disarticulate into cuboid secondary spores at maturity. In the phylogenetic analysis of Kaitsu et al. (2013), O. sessilis was resolved within a clade of Ophiocordyceps species that were later reclassified into Polycephalomycetaceae by Xiao et al. (2023). Our study further demonstrates that O. sessilis forms a strongly supported sister clade with T. zombiae (SH-aLRT = 98 %, PP = 1.0, UFB = 100 %; Fig. 3). Given the congruence of morphological traits and phylogenetic placement closely related to T. zombiae, we hereby formally transfer Ophiocordyceps sessilis to the genus Torrubiellomyces.

Ophiocordycipitaceae G.H. Sung et al., Stud. Mycol. 57: 35. 2007. MB 859522.

Notes: Ophiocordycipitaceae was introduced by Sung et al. (2007) with Ophiocordyceps as the type genus. The generic composition within this family has undergone multiple revisions (Quandt et al. 2014, Maharachchikumbura et al. 2016, Wijayawardene et al. 2018, 2020, 2022, Hyde et al. 2024). To date, Drechmeria, Hantamomyces, Harposporium, Ophiocordyceps, Paraisaria, Purpureocillium, Tolypocladium and Torrubiellomyces are accommodated in this family (Peng et al. 2024). Our multilocus phylogenetic analyses show that Torrubiellomyces falls within Polycephalomycetaceae, thereby this genus was formally excluded from Ophiocordycipitaceae (Fig. 6).

Ophiocordyceps Petch, Trans. Brit. Mycol. Soc. 16(1): 73. 1931. MB 844048.

Notes: Ophiocordyceps is the most species-rich genus within Ophiocordycipitaceae, comprising more than 348 names according to MycoBank (accession date: 25 Jun 2025). Molecular phylogenies have revealed many Ophiocordyceps species complexes that target a certain group of insect hosts and have similar morphology (Khonsanit et al. 2019, Tasanathai et al. 2020, Khao-Ngam et al. 2021, Dai et al. 2024, Ballesteros-Aguirre et al. 2025). Among them, the Ophiocordyceps pseudoacicularis species complex is composed of pathogens specific to lepidopteran larvae, producing a hirsutella-like asexual morph that was named as Hirsutella nodulosa subclade in Xie et al. (2025). Its sexual morph was distinguished by superficial, ovoid perithecia, cylindrical asci with thickened apex and filiform, whole ascospores (Tasanathai et al. 2020). In this study, we discovered a species bearing morphological characteristics similar to the Ophiocordyceps pseudoacicularis complex.

Ophiocordyceps shimenxiaensis D.P. Wei & T.C. Wen, sp. nov. MB 859523. Fig. 33 & 34.

Fig. 34 — *Ophiocordyceps shimenxiaensis* (KUNCC25-21308). **A, B.** Upper and lower view of culture on PDA medium. **C–L.** Phialides with conidia. **M–Q.** Conidia with or without mucous sheath. Scale bars: C = 50 μm; D, K–Q = 10 μm; E–J = 20 μm.

Etymology: Referring to the collection sites where the type specimen was found, shimenxia scenic spot in Ailao Mountain of Yunnan Province.

Description: Parasitic on lepidopteran larva. Asexual morph: Hirsutella-like, hyphomycetous. Phialides 9.3–20.0 ( $\overline{x} = 13.0$ , n = 20) μm long, 1.3–2.8 ( $\overline{x} = 1.8$ , n = 20) μm wide at base, 0.5–0.9 ( $\overline{x} = 0.7$ , n = 20) μm wide at apex, arising from asci released from perithecia, monophialidic, unbranched, straight, hyaline, flask-shaped, solitary, with cylindrical base and thin neck that is smooth or twisted near the apex. Conidia 3.0–6.2 × 0.9–1.5 ( $\overline{x} = 4.5 \times 1.3$ , n = 20) μm, hyaline, lemon-shaped, aseptate, singly forming at tip of phialides, without gelatinous sheath. Sexual morph: Stromata 11–15 cm long, growing from body of insect host, multiple, cylindrical, wiry, unbranched, stipitate. Stipe 2–9 cm long, 0.1–0.2 cm wide, grey, glabrous. Fertile part 0.1–0.2 cm wide, brown, cylindrical, gradually narrowing toward the apex, flexuous. Perithecia 257–527 × 158–264 ( $\overline{x} = 383 \times 224$ , n = 15) μm, ovoid, dark brown, superficial, intercalary. Peridium 24–45 ( $\overline{x} = 30$ , n = 20) μm, comprising yellow brown cells of textura angularis. Asci 135–287 × 5.0–8.2 ( $\overline{x} = 213 \times 6.9$ , n = 20) μm, cylindrical with thickened cap 4.3–7.0 × 2.3–4.4 ( $\overline{x} = 5.4 \times 3.4$ , n = 20) μm. Ascospores 140–278 × 1.6–3.4 ( $\overline{x} = 208 \times 2.1$ , n = 20) μm, hyaline, filiform, aseptate, whole.

Culture characteristics: Colonies on PDA attaining 30 mm in diam. after incubation for 30 d at 25 °C, white (1A1), felty, circular, mycelia dense, surface folded, striated with entire edge, raised at centre, with chestnut (6F7) pigment diffusing to agar, forming a transparent zone around the colony, reverse red-haired (6C4). Mycelia 3.2–5.5 ( $\overline{x} = 4.3$ , n = 30) μm, filamentous, hyaline, sparsely septate, rough-walled. Conidiophores micronematous, laterally or terminally produce alternate or opposite phialides. Phialides 32–75 ( $\overline{x} = 51$ , n = 30) μm long, mostly monophialidic or some polyphialidic, hyaline, aseptate, flask-shaped with a with distinctly swollen and verrucose base 3.7–7.5 ( $\overline{x} = 5.4$ , n = 30) μm wide, gradually tapering to a thin neck 1.1–2.0 ( $\overline{x} = 1.5$ , n = 30) μm wide, with twists in a helix at the apex, occasionally proliferation and slightly constricted at basal septum. Conidia 6–10 × 3.5–6.5 ( $\overline{x} = 8.5 \times 4.8$ , n = 45) μm, hyaline, smooth-walled, aseptate, guttulate, broadly fusiform to ovoid or lemon-shaped, arising from the apex of the neck singly, surrounded by mucous sheath, with 1–2 conidia in each mucous sheath which is broadly fusiform to globose 7.2–11.0 × 6.6–13.0 ( $\overline{x} = 9.4 \times 8.4$ , n = 30) μm. The conidia and mucous sheath become golden yellow with age.

Typus: China, Yunnan Province, Xinping County, Ailao Mountain National Reserve, on lepidopteran larva, 20 Oct. 2023, D.P. Wei, SMX10 (holotype HKAS 148572, culture ex-type KUNCC25-21308).

Additional materials examined: China, Yunnan Province, Xinping County, Ailao Mountain National Reserve, on lepidopteran larva, 20 Oct. 2023, D.P. Wei, SMX06 (HKAS 148571); ibid., SMX08 (HKAS 148569).

Notes: Multilocus phylogenetic analysis revealed limited genetic distances among species within the Ophiocordyceps pseudoacicularis complex (Fig. 6). Consequently, species delimitation in this complex relies primarily on morphological characteristics, including the colour and size of perithecia, asci, ascospores, phialides, and conidia (Tasanathai et al. 2020). The present phylogenetic tree showed that Ophiocordyceps shimenxiaensis formed a distinct clade sister to Hirsutella leigongshanensis with strong statistical support (SH-aLRT = 94 %, PP = 1, UFB = 100 %; Fig. 6). However, O. shimenxiaensis differs from H. leigongshanensis in producing a transparent zone around the colony when incubated at PDA media, a feature absent in H. leigongshanensis. Additionally, O. shimenxiaensis was found to infect lepidopteran larvae, whereas H. leigongshanensis is a parasite to coleopteran larvae. Additionally, O. shimenxiaensis produces larger phialides (32–75 × 3.7–7.5 μm vs 19–27 × 2.7–3.6 μm) and conidia (6–10 × 3.5–6.5 μm vs 4.8–6.0 × 2.4–3.6 μm) (Yu et al. 2017). Interestingly, field observations of O. shimenxiaensis showed that ascospores are forcibly ejected from mature asci in natural environments. These ascospores rapidly initiate a micro-sporulation cycle, yielding highly infectious conidia that disperse and infect new hosts. This mechanism contributes to the strong virulence and infectivity of O. shimenxiaensis, leading to high mortality rates among sympatric lepidopteran larvae. Concurrently, some O. shimenxiaensis specimens were found to be hyperparasitized by Paradingleyomyces lepidopterorum, a fungal hyperparasite that effectively suppresses host fungal growth. This interaction illustrates how hyperparasitic fungi can help maintain ecological balance between entomopathogenic fungi and their insect hosts.

Discussion

Establishment of Chlorocilliaceae and Ch. guizhouense complex

This study identified 25 hyperparasites, three allied species and three entomopathogenic fungal hosts. These hyperparasites taxonomically are distributed across nine genera (Albacillium, Calcarisporium, Chlorocillium, Fiorinimazzantia, Husseyia, Neodingleyomyces, Paralecanicillium, Paradingleyomyces, and Pleurocordyceps) within four families: Calcarisporiaceae, Chlorocilliaceae, Cordycipitaceae, and Polycephalomycetaceae. Among these, Chlorocilliaceae is a new family erected herein to accommodate Albacillium, Chlorocillium, Fiorinimazzantia, Husseyia, Neoaraneomyces, Paraneoaraneomyces, Rousseaua, Speluncomyces, and Subuliphorum. The new family is supported by both phylogenetic inference and morphological characteristics. Regarding phylogenetic analysis, a backbone tree was constructed, including taxa sampled from 28 families of Hypocreales to infer the evolutionary relationships of Chlorocilliaceae. The results show that Chlorocilliaceae constitutes a distinct lineage sister to Clavicipitaceae with strong support. Molecular dating analysis using representatives from all Hypocreales families also supported a divergence of Chlorocilliaceae from Clavicipitaceae at 142 Mya. This divergence timing allows for the introduction of a new family, according to the principle proposed by Hyde et al. (2017). The monophyly of Chlorocilliaceae and its placement as a sister lineage to Clavicipitaceae was recovered in various studies (Chen et al. 2024a, Tan & Shivas 2023a, b, 2024, Ding et al. 2024, Tan et al. 2024), further supporting its family status and the robustness of its phylogenetic placement.

Many Chlorocilliaceae species are described exclusively from their asexual morphs (Chen et al. 2019, 2024a, Ding et al. 2024), except for Chlorocillium cordycipiticola, which is known for its holomorph. Morphologically, Chlorocilliaceae displays a close relationship with Metarhizium (Clavicipitaceae) in producing green, powdery colonies on natural hosts (Mongkolsamrit et al. 2020). However, Chlorocilliaceae differs dramatically from Metarhizium by having acremonium- or hirsutella-like phialides (Zare & Gams 2016, Chen et al. 2019, 2024a, Ding et al. 2024). These phialides are commonly curved and arranged in whorls or clusters on branched conidiophores, bearing fusiform to oval to crescent-shaped conidia (Zare & Gams 2016, Chen et al. 2019, 2024a, Ding et al. 2024). In contrast, the phialides of Metarhizium are typically cylindrical, palisade-arranged, and bear oblong to cylindrical conidia (Chen et al. 2017, Mongkolsamrit et al. 2020). Additionally, Metarhizium species produce stipitate, tough, pliable stromata that are absent in Chlorocilliaceae (Mongkolsamrit et al. 2020). Members of Chlorocilliaceae instead develop superficial perithecia directly on velvety colonies (see Fig. 11). These integrated analyses strongly support Chlorocilliaceae as a distinct family separate from Clavicipitaceae. Notably, three clades were observed in Chlorocilliaceae based on the backbone tree (Fig. 3). The Clade I consist of Chlorocillium, which generates fusiform (Figs 9, 10, 12, 13–15, 17) or ovoid conidia (Figs 11, 21) in chains. Clade II comprises Albacillium, Fiorinimazzantia, Husseyia, Rousseaua, and Speluncomyces. These genera exclusively produce crescent-shaped conidia in slimy heads (Figs 22, 23, 24). Clade III contains Neoaraneomyces, Paraneoaraneomyces, and Subuliphorum that produce conidia aggregated in chain or slimy heads (Chen et al. 2022, Hou et al. 2023, Zhang et al. 2023). This distinction indicates that conidial traits are evolutionarily conserved and diagnostically significant for these specific clades. However, conidial morphology is not useful for interspecific demarcation.

Chlorocillium is the most species-rich genus within Chlorocilliaceae, comprising 22 recognized species. In this study, we added 11 novel species, one new host record (Ch. neolepidopterorum) and one reference specimen (Ch. guizhouense) to Chlorocillium. Among these, Chlorocillium guizhouense and Chlorocillium houqiaoense are pathogens of spiders, Chlorocillium coccidiicola was recorded on scale insects, and Chlorocillium tengchongense was found in lepidopteran larvae. These entomopathogenic species are closely related to hyperparasitic fungi, and their inclusion herein helps elucidate the ecological and phylogenetic connections within this group. It was found that the interspecific variation is not significant among species of Chlorocillium. Chlorocillium guizhouense complex consists of Ch. guizhouense, Ch. aschersoniae, Ch. griseum, Ch. globiceps, Ch. taiyangheens, Ch. vallense, Ch. araneogenum, and Ch. sinense. All cryptic species within this complex produce fusiform conidia with overlapping size ranges. These taxa exhibit minimal genetic distances in the various markers and minor microscopic morphological variations but demonstrate significant differences in macromorphology and host specificity. For instance, Ch. guizhouense and Ch. globiceps both produce green colonies, while the former is obligately parasitic on spider hosts, while the latter exclusively infects Ophiocordyceps cf. globices. The different host specificities among morphologically nearly identical fungi are suggestive of substrate specialization.

Biodiversity, host spectrum and trophic strategies of entomopathogens-associated hyperparasites in Hypocreales

This study substantially broadens the known host range of hyperparasitic fungi within Hypocreales. Previous reports were concentrated on Beauveria, Cordyceps, Hirsutella, Ophiocordyceps, and Paraisaria (Ban et al. 2009, Wang et al. 2014, Xiao et al. 2018, 2019, 2023, 2024, Wang et al. 2020, Araújo et al. 2022, Wei et al. 2022). We document novel associations with phylogenetically diverse entomopathogenic hosts such as Akanthomyces, Aschersonia, Ascopolyporus, Hevansia, Metarhizium, and Gibellula. We formally identified Metarhizium puerense as the host of Pleurocordyceps neoagarica, Ascopolyporus sanduensis as the host of Chlorocillium sanduense, and Ophiocordyceps shimenxiaensis as the host of Paradingleyomyces lepidopterorum through integrated morphological and phylogenetic analyses. To date, 11 genera within Clavicipitaceae, Cordycipitaceae, and Ophiocordycipitaceae have been recorded as hosts of hyperparasites (Fig. 35C, E), and a total of 63 hyperparasitic species have been described across 23 genera and six families (Clavicipitaceae, Calcarisporiaceae, Chlorocilliaceae, Cordycipitaceae, Hypocreaceae, and Polycephalomycetaceae. Polycephalomycetaceae is the most species-rich, accounting for approximately 45 % (28 species) of known hyperparasites, followed by Cordycipitaceae (25 %), Chlorocilliaceae (19 %), and Calcarisporiaceae (6 %); Clavicipitaceae and Hypocreaceae each contribute only a single recorded species (Fig. 35 A, B, D).

Fig. 35 — Bipartite network displaying reports of hyperparasitic fungi infecting entomopathogenic fungi in *Hypocreales*. **A, B.** Diversity of hyperparasitic fungi at family and species level. C. Hosts/substrates of the hyperparasites. D. Genus-level distribution patterns of hyperparasitic fungi across six families. E. Genus-level distribution patterns of entomopathogenic fungal hosts within *Clavicipitaceae*, *Cordycipitaceae*, and *Ophiocordycipitaceae*, as well as on other substrates. The data analysis was performed using OriginPro 2026, OriginLab Corporation, Northampton, MA, USA (https://www.originlab.com/).

Within this expanded taxonomic framework, hyperparasitic fungi display a spectrum of trophic strategies ranging from facultative to obligate. Many taxa (e.g., Pleurocordyceps agarica, Pl. aurantiacus, Pl. vitellina, Polycephalomyces formosus, Calcarisporium ophiocordycipiticola, Chlorocillium menghaiense) are capable of infecting multiple Ophiocordyceps species, while others (e.g., Pleurocordyceps fusiformispora, Pl. heilongtanensis, Pl. sinensis, Chlorocillium neolepidopterorum) can colonize both fungal hosts (Ophiocordyceps) and arthropod hosts, including dead insects, lepidopteran larvae and pupae, and spiders. These broad host ranges indicate facultative associations and underscore the adaptive flexibility of certain hyperparasites across diverse ecological niches. Conversely, some hyperparasites exhibit narrow host specificity. Paradingleyomyces lepidopterorum has been recorded exclusively on Ophiocordyceps shimenxiaensis across geographically separated localities in Yunnan (Tengchong City and the Ailao Mountains), suggesting an obligate association and potential co-evolutionary history (Wang et al. 2024). Additionally, a single entomopathogenic species can host multiple hyperparasites. Ophiocordyceps neogryllotalpae has been reported as host of Pleurocordyceps neoagarica, P. clavisynnema, and P. sanduensis, indicating that hyperparasite richness at the host species level can be high (Xiao et al. 2024). The trophic interface between hyperparasites and entomopathogenic fungi is further complicated by co-infection and sequential colonization dynamics. Pleurocordyceps heilongtanensis and Ophiocordyceps crinalis were found to co-occur on a single lepidopteran larva, (Fig. 28), a pattern best interpreted as sequential colonization. Ophiocordyceps initially colonizes the insect hemocoel, creating a nutrient-rich niche that is later exploited by Pleurocordyceps. Such interactions highlight hyperparasitism as a dynamic process within host-associated fungal communities. In contrast to more straightforward forms of parasitism, cryptic hyperparasitism presents significant challenges for conventional diagnostic methods. A case in point is our isolation of Albacillium hingganense from asymptomatic sclerotia of an ant-parasitizing Ophiocordyceps species. Similarly, Leptobacillium hepiali was also isolated from the sclerotia of an Ophiocordyceps species (Shen et al. 2026). Such instances suggest that cryptic hyperparasites may exist in a subclinical or latent state within their hosts, possibly undergoing an endophytic-like phase prior to transitioning to a more tropically active form. These findings raise important questions about the complex and often hidden dynamics of hyperparasitism, emphasizing the need for more advanced diagnostic approaches to uncover the full range of interactions between parasitic fungi and their hosts.

Physiological impacts and ecological significance of hyperparasites

Hyperparasitic fungi function as a critical regulatory force within ecosystems, exerting top-down control on entomopathogenic fungi (Bermúdez-Cova et al. 2023). The complexity of these interactions is exemplified by the discovery of hyperparasitic genera such as Niveomyces and Torrubiellomyces on the zombie-ant fungus Ophiocordyceps camponoti-floridani, which can drastically reduce host fitness by suppressing or distorting stroma formation (Araújo et al. 2022). Although empirical data linking hyperparasite prevalence to entomopathogenic fungal density and insect population dynamics remain scarce, current evidence suggests that hyperparasites likely act as modulators of disease epizootics and broader trophic cascades, thereby helping to maintain ecosystem equilibrium (Andersen et al. 2012). This regulatory role is achieved through specialized physiological mechanisms. Hyperparasitic fungi exhibit considerable morphological plasticity in their colonization strategies, ranging from conspicuous macroscopic colonies on fertile heads, stipes, and external insect bodies to cryptic lifestyles involving the parasitism of minute synnemata (Fig. 36A, B). Infection typically begins with adhesion to host surfaces, followed by tissue penetration and the frequent development of specialized feeding structures, such as hooks, for nutrient extraction (Sun et al. 2019). Concurrently, hyperparasitic fungi employ chemical warfare by secreting antifungal metabolites like harzianic acid (Sawa et al. 1994, El-Hasan et al. 2022). This combination of physical invasion and chemical inhibition frequently results in the effective castration of the primary pathogen, whereby hyperparasitic fungi suppress or sterilize reproductive structures, severely compromising the fitness of the entomopathogenic fungal host (Aneja et al. 2005). For instance, this study found that Pleurocordyceps heilongtanensis and P. neoagarica render host conidia non-viable despite their normal appearance. Furthermore, Chlorocillium cordycipiticola, Ch. globiceps, Ch. puerense, Husseyia ramosissima, Paralecanicillium yunnanense, and Neodingleyomyces tengchongensis can completely overgrow and degrade host mycelia, erasing diagnostic features and accelerating DNA degradation. These processes will complicate species-level identification of entomopathogenic fungal hosts in the absence of co-occurring intact specimens.

Schematic representation of the interactions between hyperparasites (HPFs) and entomopathogenic fungi (EMPs) within a natural ecosystem. A. Morphological structure of EMPs, showcasing macroscopic colonies of HPFs on the fertile head, stipe, and insect host. B. Synnemata of *Ophiocordyceps* sp. infected by the hyperparasite, highlighting the phialides and conidia of both the hyperparasite and its fungal host. C. Diversity of ecological niches of hyperparasites and their allied species, illustrating the ecological interactions between HPFs, EMPs, and insect hosts. The icons and templates used in Fig. 36C were cited from BioRender’s library (BioRender.com).

Ecologically, the influence of hyperparasitic fungi extends far beyond simple parasitism. By castrating or decomposing entomopathogenic fungi, they indirectly promote insect survival, adding a regulatory layer to tritrophic interactions (Andersen et al. 2012, Bermúdez-Cova et al. 2023). Furthermore, hyperparasitic fungi act as ecosystem engineers by decomposing fungal biomass and accelerating nutrient turnover in soil and leaf litter. Their ecological reach is amplified by diverse environmental reservoirs. Isolates closely related to known hyperparasites have been recovered from insect cadavers, soil, and endosclerotia (Fig. 36C). This suggests that saprobic or endophytic phases in non-fungal substrates facilitate dispersal, embedding HPFs within complex interaction networks that bridge different functional guilds and habitat types.

Acknowledgments

This work was jointly supported by the National Natural Science Foundation of China (No. 32570025) and Guizhou Provincial Scientist Workstation (No.KXJZ [2025]002).

Footnotes

DATA AVAILABILITY

The sequences generated in this study are deposited with NCBI GenBank (See Tables S1–S5). The alignments and phylogenetic trees used for Figs 3–7 were submitted to Figshare (doi: https://doi.org/10.6084/m9.figshare.31627150).

DECLARATION ON CONFLICT OF INTEREST

The authors declare that there is no conflict of interest.

Supplementary Material: https://studiesinmycology.org/

Table S1

Taxa and GenBank accession numbers used in the phylogenetic analysis of Hypocreales. The newly generated isolates/sequences are indicated in black bold. Type strains/species are marked with superscript ‘T’.

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S2

Taxa and GenBank accession numbers used in the phylogenetic analysis of Chlorocillium. The newly generated isolates/sequences are indicated in black bold. Type strains/species are marked with superscript ‘T’.

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S3

Taxa and GenBank accession numbers used in the phylogenetic analysis of Pleurocordyceps. The newly generated isolates/sequences are indicated in black bold. Type strains/species are marked with superscript ‘T’.

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S4

Taxa and GenBank accession numbers used in the phylogenetic analysis of Ophiocordyceps. The newly generated isolates/sequences are indicated in black bold. Type strains/species are marked with superscript ‘T’.

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S5

Taxa and GenBank accession numbers used in molecular dating analysis of families in Hypocreales. The newly generated isolates/sequences are indicated in black bold. Type strains/species are marked with superscript ‘T’.

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

References

Alves VC, Lira RA, Lima JM, et al. (2022). Unravelling the fungal darkness in a tropical cave: richness and the description of one new genus and six new species. Fungal Systematics and Evolution 10(1): 139–167. 10.3114/fuse.2022.10.06 [DOI] [PMC free article] [PubMed] [Google Scholar]
Andersen SB, Ferrari M, Evans HC, et al. (2012). Disease dynamics in a specialized parasite of ant societies. PloS ONE 7(5): e36352. 10.1371/journal.pone.0036352 [DOI] [PMC free article] [PubMed] [Google Scholar]
Aneja M, Gianfagna TJ, Hebbar PK. (2005). Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens. Physiological and Molecular Plant Pathology 67(6): 304–307. 10.1016/j.pmpp.2006.05.002 [DOI] [Google Scholar]
Araújo JPM, Lebert BM, Vermeulen S, et al. (2022). Masters of the manipulator: two new hypocrealean genera, Niveomyces (Cordycipitaceae) and Torrubiellomyces (Ophiocordycipitaceae), parasitic on the zombie ant fungus Ophiocordyceps camponoti-floridani. Persoonia 49(1): 171–194. 10.3767/persoonia.2022.49.05 [DOI] [PMC free article] [PubMed] [Google Scholar]
Ballesteros-Aguirre CE, Sanjuan T, Ramírez-Cruz V, et al. (2025). Zombie-ant fungi from western Mexico: six new species in the Ophiocordyceps unilateralis complex (Hypocreales: Ascomycota) and a new host association with Cephalotes ants. Persoonia 55(1): 203–238. 10.3114/persoonia.2025.55.06 [DOI] [PMC free article] [PubMed] [Google Scholar]
Ban S, Sakane T, Toyama K, et al. (2009). Teleomorph–anamorph relationships and reclassification of Cordyceps cuboidea and its allied species. Mycoscience 50(4): 261–272. 10.1007/S10267-008-0480-Y [DOI] [Google Scholar]
Bandelt HJ, Dress AWM. (1992). Split decomposition: a new and useful approach to phylogenetic analysis of distance data. Molecular Phylogenetics and Evolution 1(3): 242–252. 10.1016/1055-7903(92)90021-8 [DOI] [PubMed] [Google Scholar]
Bao DF, Hyde KD, Maharachchikumbura SS, et al. (2023). Taxonomy, phylogeny and evolution of freshwater Hypocreomycetidae (Sordariomycetes). Fungal Diversity 121(1): 1–94. 10.1007/s13225-023-00521-8 [DOI] [Google Scholar]
Berbee ML, Taylor JW. (2010). Dating the molecular clock in fungi – how close are we? Fungal Biology Reviews 24(1–2): 1–16. 10.1016/j.fbr.2010.03.001 [DOI] [Google Scholar]
Bermúdez-Cova MA, Haelewaters D, de Bekker C, et al. (2023). Hyperparasitic fungi—definitions, diversity, ecology, and research. Authorea. 10.22541/au.168787020.07281183/v2 [DOI] [Google Scholar]
Bischoff JF, Chaverri P, White JF., Jr (2005). Clarification of the host substrate of Ascopolyporus and description of Ascopolyporus philodendrus sp. nov. Mycologia 97(3): 710–717. 10.3852/mycologia.97.3.710 [DOI] [PubMed] [Google Scholar]
Candoussau F, Boqueras M, Gómez-Bolea A, et al. (2007). Observations on Neobarya, including new species and new combinations. Sydowia 59(2): 179–215. [Google Scholar]
Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T. (2009). trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15): 1972–1973. 10.1093/bioinformatics/btp348 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chaverri P, Liu M, Hodge KT. (2008). A monograph of the entomopathogenic genera Hypocrella, Moelleriella, and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their aschersonia-like anamorphs in the Neotropics. Studies in Mycology 60: 1–66. 10.3114/sim.2008.60.01 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chen WH, Han YF, Liang JD, et al. (2017). Metarhizium dendrolimatilis, a novel Metarhizium species parasitic on Dendrolimus sp. larvae. Mycosphere 8: 31–37. 10.5943/mycosphere/8/1/4 [DOI] [Google Scholar]
Chen WH, Han YF, Liang JD, et al. (2019). Morphological and phylogenetic characterization of novel Metarhizium species in Guizhou, China. Phytotaxa 419(2): 189–196. 10.11646/phytotaxa.419.2.5 [DOI] [Google Scholar]
Chen WH, Li D, Liang J D, et al. (2024b). Two new Cordyceps-like species, Perennicordyceps zongqii sp. nov. (Polycephalomycetaceae) and Purpureocillium zongqii sp. nov. (Ophiocordycipitaceae), in Hypocreales from karst region of China. MycoKeys 110: 141–158. 10.3897/mycokeys.110.135724 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chen WH, Li D, Liang JD, et al. (2024a). Chlorocillium sinense sp. nov. (Clavicipitaceae) and Calcarisporium guizhouense sp. nov. (Calcarisporiaceae) in Hypocreales from China. MycoKeys 109: 91–107. 10.3897/mycokeys.109.128060 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chen WH, Li D, Shu HL, et al. (2025a). Four new araneogenous species and a new genus in Hypocreales (Clavicipitaceae, Cordycipitaceae) from the karst region of China. MycoKeys 112: 335–359. 10.3897/mycokeys.112.140799 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chen WH, Liang JD, Ren XX, et al. (2022). Phylogenetic, ecological and morphological characteristics reveal two new spider-associated genera in Clavicipitaceae. MycoKeys 91: 49–66. 10.3897/mycokeys.91.86812 [DOI] [PMC free article] [PubMed] [Google Scholar]
Chen WH, Shu HL, Li D, et al. (2025b). Shedding light on the darkness: Cryptic diversity of cordyceps-like fungi in karst regions of Guizhou Province, China. Mycosphere 16: 2887–2974. 10.5943/mycosphere/16/1/20 [DOI] [Google Scholar]
Chernomor O, Von Haeseler A, Minh BQ. (2016). Terrace aware data structure for phylogenomic inference from supermatrices. Systematic Biology 65(6): 997–1008. 10.1093/sysbio/syw037 [DOI] [PMC free article] [PubMed] [Google Scholar]
Clewley JP. (1995). Macintosh sequence analysis software. Molecular Biotechnology 3: 221–224. 10.1007/BF02789332 [DOI] [PubMed] [Google Scholar]
Crous PW, Costa MM, Kandemir H, et al. (2023). Fungal Planet description sheets: 1550–1613. Persoonia 51: 280–417. 10.3767/persoonia.2023.51.08 [DOI] [PMC free article] [PubMed] [Google Scholar]
Crous PW, Gams W, Stalpers JA, et al. (2004). MycoBank: an online initiative to launch mycology into the 21st century. Studies in Mycology 50: 19–22. [Google Scholar]
Custódio FA, Pereira OL. (2025). New treasures in Cordycipitaceae: fungicolous fungi associated with Pseudocercospora fijiensis and P. musae in Brazil, including Matutinistella gen. nov. Fungal Systematics and Evolution 15(1): 133–152. 10.3114/fuse.2025.15.06 [DOI] [PMC free article] [PubMed] [Google Scholar]
Dai Y, Chen S, Wang Y, et al. (2024). Molecular phylogenetics of the Ophiocordyceps sinensis-species complex lineage (Ascomycota, Hypocreales), with the discovery of new species and predictions of species distribution. IMA Fungus 15(1): 2. 10.1186/s43008-023-00131-8 [DOI] [PMC free article] [PubMed] [Google Scholar]
Darmostuk V, Etayo J, Rodriguez-Flakus P, et al. (2025). A novel, exclusively lichen-inhabiting lineage of hypocrealean fungi revealed in the Sordariomycetes. Persoonia 54: 47–91. 10.3114/persoonia.2025.54.02 [DOI] [PMC free article] [PubMed] [Google Scholar]
Ding M, Li Y, Bai Y, et al. (2024). Albacillium hingganense gen. et sp. nov. (Clavicipitaceae) from forest litters in Northeast China. Phytotaxa 650(2): 157–168. 10.11646/phytotaxa.650.2.3 [DOI] [Google Scholar]
Dong W, Hyde KD, Jeewon R, et al. (2025). Fungal diversity notes 2017–2122: taxonomic and phylogenetic contributions to freshwater fungi and other fungal taxa. Fungal Diversity 134: 185–459. 10.1007/s13225-025-00560-3 [DOI] [Google Scholar]
El-Hasan A, Walker F, Klaiber I, et al. (2022). New approaches to manage Asian soybean rust (Phakopsora pachyrhizi) using Trichoderma spp. or their antifungal secondary metabolites. Metabolites 12(6): 507. 10.3390/metabo12060507 [DOI] [PMC free article] [PubMed] [Google Scholar]
Flakus A, Etayo J, Miadlikowska J, et al. (2019). Biodiversity assessment of ascomycetes inhabiting Lobariella lichens in Andean cloud forests led to one new family, three new genera and 13 new species of lichenicolous fungi. Plant and Fungal Systematics 64(2): 283–344. 10.2478/pfs-2019-0022 [DOI] [Google Scholar]
Gams W. (1971). Cephalosporium-artige Schimmelpilze (Hyphomycetes). Gustav Fischer Verlag, Stuttgart. [Google Scholar]
Gonzalez MC, Glenn AE, Hanlin RT, et al. (2016). Acremonium camptosporum isolated as an endophyte of Bursera simaruba from Yucatan Peninsula, Mexico. Mycotaxon 131(1): 211–225. [Google Scholar]
Guindon S, Dufayard JF, Lefort V, et al. (2010). New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic Biology 59(3): 307–321. 10.1093/sysbio/syq010 [DOI] [PubMed] [Google Scholar]
Hall TA. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98. [Google Scholar]
Hasan R, Lv B, Uddin MJ, et al. (2022). Monitoring mycoparasitism of Clonostachys rosea against Botrytis cinerea using GFP. Journal of Fungi 8(6): 567. 10.3390/jof8060567 [DOI] [PMC free article] [PubMed] [Google Scholar]
Hennings P. (1902). Fungi blumenaevienses II. a cl. Alfr. Möller lecti. Hedwigia 41(1): 1–33. [Google Scholar]
Hennings P. (1909). Fungi S. Paulensis IV a cl. Puttemans collecti. Hedwigia 48: 1–20. [Google Scholar]
Hongsanan S, Maharachchikumbura SSN, Hyde KD, et al. (2017). An updated phylogeny of Sordariomycetes based on phylogenetic and molecular clock evidence. Fungal Diversity 84(1): 25–41. 10.1007/s13225-017-0384-2 [DOI] [Google Scholar]
Hoog GS. (1978). Notes on some fungicolous hyphomycetes and their relatives. Persoonia 10(1): 33–81. [Google Scholar]
Hou LW, Giraldo A, Groenewald JZ, et al. (2023). Redisposition of acremonium-like fungi in Hypocreales. Studies in Mycology 105: 23–203. 10.3114/sim.2023.105.02 [DOI] [PMC free article] [PubMed] [Google Scholar]
Huson DH, Bryant D. (2024). The SplitsTree app: interactive analysis and visualization using phylogenetic trees and networks. Nature Methods 21(10): 1773–1774. 10.1038/s41592-024-02406-3 [DOI] [PubMed] [Google Scholar]
Hyde KD, Noorabadi M T, Thiyagaraja V, et al. (2024). The 2024 outline of Fungi and fungus-like taxa. Mycosphere 15(1): 5146–6239. 10.5943/mycosphere/15/1/25 [DOI] [Google Scholar]
Hyde KD, Maharachchikumbura SS, Hongsanan S, et al. (2017). The ranking of fungi: a tribute to David L. Hawksworth on his 70th birthday. Fungal Diversity 84: 1–23. 10.1007/s13225-017-0383-3 [DOI] [Google Scholar]
Johnston PR, Park D. (2023). Dingleyomyces lloydii (Hypocreales, Polycephalomycetaceae), gen. nov., comb. nov. New Zealand Journal of Botany 63(4): 851–858. 10.1080/0028825X.2023.2291453 [DOI] [Google Scholar]
Kaitsu Y, Shimizu K, Tanaka E, et al. (2013). Ophiocordyceps sessilis sp. nov., a new species of Ophiocordyceps on Camponotus ants in Japan. Mycological Progress 12: 755–761. 10.1007/s11557-013-0911-9 [DOI] [Google Scholar]
Kalyaanamoorthy S, Minh BQ, Wong TK, et al. (2017). ModelFinder: fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. 10.1038/nmeth.4285 [DOI] [PMC free article] [PubMed] [Google Scholar]
Katoh K, Rozewicki J, Yamada KD. (2019). MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20: 1160–1166. 10.1093/bib/bbx108 [DOI] [PMC free article] [PubMed] [Google Scholar]
Kepler RM, Sung GH, Harada Y, et al. (2012). Host jumping onto close relatives and across kingdoms by Tyrannicordyceps (Clavicipitaceae) gen. nov. and Ustilaginoidea (Clavicipitaceae). American Journal of Botany 99(3): 552–561. 10.3732/ajb.1100124 [DOI] [PubMed] [Google Scholar]
Kepler RM, Luangsa-Ard JJ, Hywel-Jones NL, et al. (2017). A phylogenetically-based nomenclature for Cordycipitaceae (Hypocreales). IMA Fungus 8(2): 335–353. 10.5598/imafungus.2017.08.02.08 [DOI] [PMC free article] [PubMed] [Google Scholar]
Khao-Ngam S, Mongkolsamrit S, Rungjindamai N, et al. (2021). Ophiocordyceps asiana and Ophiocordyceps tessaratomidarum (Ophiocordycipitaceae, Hypocreales), two new species on stink bugs from Thailand. Mycological Progress 20: 341–353. 10.1007/s11557-021-01684-x [DOI] [Google Scholar]
Khonsanit A, Luangsa-Ard JJ, Thanakitpipattana D, et al. (2019). Cryptic species within Ophiocordyceps myrmecophila complex on formicine ants from Thailand. Mycological Progress 18: 147–161. 10.1007/s11557-018-1412-7 [DOI] [Google Scholar]
Khonsanit A, Thanakitpipattana D, Mongkolsamrit S, et al. (2024). A phylogenetic assessment of Akanthomyces sensu lato in Cordycipitaceae (Hypocreales, Sordariomycetes): introduction of new genera, and the resurrection of Lecanicillium. Fungal Systematics and Evolution 14(1): 271–306. 10.3114/fuse.2024.14.17 [DOI] [PMC free article] [PubMed] [Google Scholar]
Kobmoo N, Mongkolsamrit S, Khonsanit A, et al. (2024). Integrative taxonomy of Metarhizium anisopliae species complex, based on phylogenomics combined with morphometrics, metabolomics, and virulence data. IMA Fungus 15(1): 30. 10.1186/s43008-024-00154-9 [DOI] [PMC free article] [PubMed] [Google Scholar]
Kobmoo N, Tasanathai K, Araújo JP, et al. (2023). New mycoparasitic species in the genera Niveomyces and Pseudoniveomyces gen. nov. (Hypocreales: Cordycipitaceae), with sporothrix-like asexual morphs, from Thailand. Fungal Systematics and Evolution 12(1): 91–110. 10.3114/fuse.2023.12.07 [DOI] [PMC free article] [PubMed] [Google Scholar]
Kornerup A, Wanscher JH. (1978). Methuen Handbook of Colour, 3rd edn. Eyre Methuen Ltd., London. [Google Scholar]
Kryukov V, Yaroslavtseva O, Tyurin M, et al. (2017). Ecological preferences of Metarhizium spp. from Russia and neighboring territories and their activity against Colorado potato beetle larvae. Journal of Invertebrate Pathology 149: 1–7. 10.1016/j.jip.2017.07.001 [DOI] [PubMed] [Google Scholar]
Kuraku S, Zmasek CM, Nishimura O, et al. (2013). aLeaves facilitates on-demand exploration of metazoan gene family trees on MAFFT sequence alignment server with enhanced interactivity. Nucleic Acids Research 41: W22–W28. 10.1093/nar/gkt389 [DOI] [PMC free article] [PubMed] [Google Scholar]
Larsson A. (2014). AliView: a fast and lightweight alignment viewer and editor for large datasets. Bioinformatics 30(22): 3276–3278. 10.1093/bioinformatics/btu531 [DOI] [PMC free article] [PubMed] [Google Scholar]
Lee K, Missaoui A, Mahmud K, et al. (2021). Interaction between grasses and Epichloë endophytes and its significance to biotic and abiotic stress tolerance and the rhizosphere. Microorganisms 9(11): 2186. 10.3390/microorganisms9112186 [DOI] [PMC free article] [PubMed] [Google Scholar]
Lin L, Lin Y, Keyhani NO, et al. (2025). New entomopathogenic species in the Clavicipitaceae family (Hypocreales, Ascomycota) from the subtropical forests of Fujian, China. Frontiers in Microbiology 16: 1532341. 10.3389/fmicb.2025.1532341 [DOI] [PMC free article] [PubMed] [Google Scholar]
Liu M, Shoukouhi P, Bisson K R, et al. (2021b). Sympatric divergence of the ergot fungus, Claviceps purpurea, populations infecting agricultural and nonagricultural grasses in North America. Ecology and Evolution 11(1): 273–293. 10.1002/ece3.7028. [DOI] [PMC free article] [PubMed] [Google Scholar]
Liu Q, Dong C. (2023). Dual transcriptomics reveals interspecific interactions between the mycoparasite Calcarisporium cordycipiticola and its host Cordyceps militaris. Microbiology Spectrum 11(2): e04800-22. 10.1128/spectrum.04800-22 [DOI] [PMC free article] [PubMed] [Google Scholar]
Liu Q, Wang F, Xu FX, et al. (2021a). Pathogenic fungi of artificially cultivated Cordyceps militaris. Mycosystema 40(11): 2962–2980. 10.13346/j.mycosystema.210147 [DOI] [Google Scholar]
Liu Z, Tang D, Lu Y, et al. (2024). Morphology and phylogeny of four new species within Polycephalomycetaceae (Hypocreales) parasitising Ophiocordyceps species. MycoKeys 105: 179–202. 10.3897/mycokeys.105.119893 [DOI] [PMC free article] [PubMed] [Google Scholar]
Ma JM, Wang ZQ, Yang ZL, et al. (2024). Metarhizium puerense (Hypocreales, Clavicipitaceae): a new species from Yunnan, south-western China. Biodiversity Data Journal 12: e129087. 10.3897/BDJ.12.e129087 [DOI] [PMC free article] [PubMed] [Google Scholar]
Maharachchikumbura SSN, Hyde KD, Jones EBG, et al. (2016). Families of Sordariomycetes. Fungal Diversity 79: 1–317. 10.1007/s13225-016-0369-6 [DOI] [Google Scholar]
Matsushima T. (1975). Icones Microfungorum a Matsushima Lectorum. Published by the author. Kobe, Japan. [Google Scholar]
Minh BQ, Nguyen MA, Von Haeseler A. (2013). Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution 30(5): 1188–1195. 10.1093/molbev/mst024 [DOI] [PMC free article] [PubMed] [Google Scholar]
Möller A. (1901). Phycomyceten und Ascomyceten. Untersuchungen aus Brasilien. Botanische Mittheilungen aus den Tropen 9: 1–319. 10.5962/bhl.title.2753 [DOI] [Google Scholar]
Mongkolsamrit S, Khonsanit A, Thanakitpipattana D, et al. (2020). Revisiting Metarhizium and the description of new species from Thailand. Studies in Mycology 95(1): 171–251. 10.1016/j.simyco.2020.04.001 [DOI] [PMC free article] [PubMed] [Google Scholar]
Mongkolsamrit S, Noisripoom W, Tasanathai K, et al. (2022). Comprehensive treatise of Hevansia and three new genera Jenniferia, Parahevansia and Polystromomyces on spiders in Cordycipitaceae from Thailand. MycoKeys 91: 113–145. 10.3897/mycokeys.91.83091 [DOI] [PMC free article] [PubMed] [Google Scholar]
Mongkolsamrit S, Noisripoom W, Thanakitpipattana D, et al. (2018). Disentangling cryptic species with Isaria-like morphs in Cordycipitaceae. Mycologia 110(1): 230–257. 10.1080/00275514.2018.1446651 [DOI] [PubMed] [Google Scholar]
Mongkolsamrit S, Sandargo B, Ebada SS, et al. (2023). Bhushaniella gen. nov. (Cordycipitaceae) on spider eggs sac: a new genus from Thailand and its bioactive secondary metabolites. Mycological Progress 22(9): 64. 10.1007/s11557-023-01915-3 [DOI] [Google Scholar]
Mukherjee PK, Mendoza-Mendoza A, Zeilinger S, et al. (2022). Mycoparasitism as a mechanism of Trichoderma-mediated suppression of plant diseases. Fungal Biology Reviews 39: 15–33. 10.1016/j.fbr.2021.11.004 [DOI] [Google Scholar]
Nguyen LT, Schmidt HA, Von Haeseler A, et al. (2015). IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268–274. 10.1093/molbev/msu300 [DOI] [PMC free article] [PubMed] [Google Scholar]
Nguyen TT, Le TN, Nguyen TH. (2023). First report of emerging fungal pathogens of Cordyceps militaris in Vietnam. Scientific Reports 13(1): 17669. 10.1038/s41598-023-43951-9 [DOI] [PMC free article] [PubMed] [Google Scholar]
Nylander JAA. (2004). MrModeltest 2.2: program distributed by the author. Evolutionary Biology Centre, Uppsala University. [Google Scholar]
Peng XC, Wen TC, Wei DP, et al. (2024). Two new species and one new combination of Ophiocordyceps (Hypocreales, Ophiocordycipitaceae) in Guizhou. MycoKeys 102: 245–266. 10.3897/mycokeys.102.113351 [DOI] [PMC free article] [PubMed] [Google Scholar]
Petch T. (1926). Entomogenous fungi: additions and corrections. II. Transactions of the British Mycological Society 11(3–4): 258–266. 10.1016/S0007-1536(26)80010-5 [DOI] [Google Scholar]
Preuss CGT. (1851). Synopsis fungorum hucusque cognitorum praesertim prope Hoyerswerda. Linnaea 24: 101–153. [Google Scholar]
Quandt CA, Kepler RM, Gams W, et al. (2014). Phylogenetic-based nomenclatural proposals for Ophiocordycipitaceae (Hypocreales) with new combinations in Tolypocladium. IMA Fungus 5: 121–134. 10.5598/imafungus.2014.05.01.12 [DOI] [PMC free article] [PubMed] [Google Scholar]
Rambaut A. (2018). FigTree v. 1.4.4. Available online: http://tree.bio.ed.ac.uk/software/figtree/.
Rambaut A, Suchard MA, Xie D, et al. (2013). Tracer version 1.6. University of Edinburgh. [Google Scholar]
Ronquist F, Teslenko M, van der Mark P, et al. (2012). MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systems Biology 61: 539–542. 10.1093/sysbio/sys029 [DOI] [PMC free article] [PubMed] [Google Scholar]
Samarakoon MC, Hyde KD, Maharachchikumbura SSM, et al. (2022). Taxonomy, phylogeny, molecular dating and ancestral state reconstruction of Xylariomycetidae (Sordariomycetes). Fungal Diversity 112(1): 1–88. 10.1007/s13225-021-00495-5 [DOI] [Google Scholar]
Sawa R, Mori Y, Iinuma H, et al. (1994). Harzianic acid, a new antimicrobial antibiotic from a fungus. The Journal of Antibiotics 47(6): 731–732. 10.7164/antibiotics.47.731 [DOI] [PubMed] [Google Scholar]
Shen J, Gou W, Tian HL, et al. (2026). Molecular phylogeny and morphology reveal two new species, Ophiocordyceps ramosa sp. nov. (Ophiocordycipitaceae) and Leptobacillium hepiali sp. nov. (Cordycipitaceae), in Hypocreales from China. MycoKeys 127: 191. 10.3897/mycokeys.127.173361 [DOI] [PMC free article] [PubMed] [Google Scholar]
Somrithipol S, Jones EBG. (2006). Calcarisporium phaeopodium sp. nov., a new hyphomycete from Thailand. Sydowia 58(1): 133–140. [Google Scholar]
Steel MA. (1994). Recovering a tree from the leaf colourations it generates under a Markov model. Applied Mathematics Letters 7(2): 19–24. 10.1016/0893-9659(94)90024-8 [DOI] [Google Scholar]
Subramanian CV. (1978). Revisions of Hyphomycetes I. Kavaka 5: 93–98. [Google Scholar]
Sun JZ, Dong CH, Liu XZ, et al. (2016). Calcarisporium cordycipiticola sp. nov., an important fungal pathogen of Cordyceps militaris. Phytotaxa 268(2): 135–144. 10.11646/phytotaxa.268.2.4 [DOI] [Google Scholar]
Sun JZ, Liu XZ, Hyde KD, et al. (2017). Calcarisporium xylariicola sp. nov. and introduction of Calcarisporiaceae fam. nov. in Hypocreales. Mycological Progress 16(4): 433–445. 10.1007/s11557-017-1290-4 [DOI] [Google Scholar]
Sun JZ, Liu XZ, McKenzie EH, et al. (2019). Fungicolous fungi: terminology, diversity, distribution, evolution, and species checklist. Fungal Diversity 95: 337–430. 10.1007/s13225-019-00422-9 [DOI] [Google Scholar]
Sung GH, Hywel-Jones NL, Sung JM, et al. (2007). Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Studies in Mycology 57: 5–59. 10.3114/sim.2007.57.01 [DOI] [PMC free article] [PubMed] [Google Scholar]
Sung GH, Poinar GO, Jr, Spatafora JW. (2008). The oldest fossil evidence of animal parasitism by fungi supports a Cretaceous diversification of fungal–arthropod symbioses. Molecular Phylogenetics and Evolution 49: 495–502. 10.1016/j.ympev.2008.08.028 [DOI] [PubMed] [Google Scholar]
Tan TP, Valter S, Lacey E. (2025). Nomenclatural novelties. Index of Australian Fungi 62: 1–20. 10.5281/zenodo.17336362 [DOI] [Google Scholar]
Tan YP, Minns SA, Valter S, et al. (2024). Nomenclatural novelties. Index of Australian Fungi 47: 1–14. 10.5281/zenodo.14177440 [DOI] [Google Scholar]
Tan YP, Shivas RG. (2023a). Nomenclatural novelties. Index of Australian Fungi 22: 1–14. 10.5281/zenodo.10145676 [DOI] [Google Scholar]
Tan YP, Shivas RG. (2023b). Nomenclatural novelties. Index of Australian Fungi 27: 1–18. 10.5281/zenodo.10426579 [DOI] [Google Scholar]
Tan YP, Shivas RG. (2024b). Nomenclatural novelties. Index of Australian Fungi 31: 1–12. 10.5281/zenodo.10784071 [DOI] [Google Scholar]
Tasanathai K, Thanakitpipattana D, Himaman W, et al. (2020). Three new Ophiocordyceps species in the Ophiocordyceps pseudoacicularis species complex on Lepidoptera larvae in Southeast Asia. Mycological Progress 19(10): 1043–1056. 10.1007/s11557-020-01611-6 [DOI] [Google Scholar]
Thanakitpipattana D, Mongkolsamrit S, Khonsanit A, et al. (2022). Is Hyperdermium congeneric with Ascopolyporus? Phylogenetic relationships of Ascopolyporus spp. (Cordycipitaceae, Hypocreales) and a new genus Neohyperdermium on scale insects in Thailand. Journal of Fungi 8(5): 516. 10.3390/jof8050516 [DOI] [PMC free article] [PubMed] [Google Scholar]
Vaidya G, Lohman DJ, Meier R. (2011). SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27: 171–180. 10.1111/j.1096-0031.2010.00329.x [DOI] [PubMed] [Google Scholar]
Wang L, Li HH, Chen YQ, et al. (2014). Polycephalomyces lianzhouensis sp. nov., a new species, co-occurs with Ophiocordyceps crinalis. Mycological Progress 13: 1089–1096. 10.1007/s11557-014-0996-9 [DOI] [Google Scholar]
Wang Y, Wei D P, Peng X C, et al. (2024). Interesting mycoparasites and Paradingleyomyces lepidopterorum gen. et sp. nov. (Hypocreales, Polycephalomycetaceae) from Yunnan Province, China. MycoKeys 110: 185–210. 10.3897/mycokeys.110.134132 [DOI] [PMC free article] [PubMed] [Google Scholar]
Wang YB, Wang Y, Fan Q, et al. (2020). Multigene phylogeny of the family Cordycipitaceae (Hypocreales): new taxa and the new systematic position of the Chinese cordycipitoid fungus Paecilomyces hepiali. Fungal Diversity 103: 1–46. 10.1007/s13225-020-00457-3 [DOI] [Google Scholar]
Wang YH, Ban S, Wang WJ, et al. (2021). Pleurocordyceps gen. nov. for a clade of fungi previously included in Polycephalomyces based on molecular phylogeny and morphology. Journal of Systematics and Evolution 59(5): 1065–1080. 10.1111/jse.12705 [DOI] [Google Scholar]
Wei DP, Gentekaki E, Wanasinghe DN, et al. (2022). Diversity, molecular dating and ancestral characters state reconstruction of entomopathogenic fungi in Hypocreales. Mycosphere 13(2): 281–351. 10.5943/mycosphere/si/1f/8 [DOI] [Google Scholar]
Wei DP, Wanasinghe DN, Hyde KD, et al. (2019). The genus Simplicillium. MycoKeys 60: 69. 10.3897/mycokeys.60.38040 [DOI] [PMC free article] [PubMed] [Google Scholar]
Wei DP, Wanasinghe DN, Hyde KD, et al. (2020). Ophiocordyceps tianshanensis sp. nov. on ants from Tianshan mountains, P. R. China. Phytotaxa 464(4): 277–292. 10.11646/phytotaxa.464.4.2 [DOI] [Google Scholar]
Wei DP, Wanasinghe DN, Xu JC, et al. (2021). Three novel entomopathogenic fungi from China and Thailand. Frontiers in Microbiology 11: 608991. 10.3389/fmicb.2020.608991 [DOI] [PMC free article] [PubMed] [Google Scholar]
Wen TC, Xiao YP, Han YF, et al. (2017). Multigene phylogeny and morphology reveal that the Chinese medicinal mushroom ‘Cordyceps gunnii’ is Metacordyceps neogunnii sp. nov. Phytotaxa 302(1): 27–39. 10.11646/phytotaxa.302.1.2 [DOI] [Google Scholar]
Wijayawardene NN, Hyde KD, Al-Ani LKT, et al. (2020). Outline of Fungi and fungus-like taxa. Mycosphere 11: 1060–1456. 10.5943/mycosphere/11/1/8 [DOI] [Google Scholar]
Wijayawardene NN, Hyde KD, Dai DQ, et al. (2022). Outline of Fungi and fungus-like taxa – 2021. Mycosphere 13(1): 53–453. 10.5943/mycosphere/13/1/2 [DOI] [Google Scholar]
Wijayawardene NN, Hyde KD, Lumbsch HT, et al. (2018). Outline of Ascomycota: 2017. Fungal Diversity 88: 167–263. 10.1007/s13225-018-0394-8 [DOI] [Google Scholar]
Xiang CY, Gao F, Jakovlić I. et al. (2023). Using PhyloSuite for molecular phylogeny and tree‐based analyses. iMeta 1: e87. 10.1002/imt2.87 [DOI] [PMC free article] [PubMed] [Google Scholar]
Xiao YP, Hongsanan S, Hyde KD, et al. (2019). Two new entomopathogenic species of Ophiocordyceps in Thailand. MycoKeys 47: 53–74. 10.3897/mycokeys.47.29898 [DOI] [PMC free article] [PubMed] [Google Scholar]
Xiao YP, Wang YB, Hyde KD, et al. (2023). Polycephalomycetaceae, a new family of clavicipitoid fungi segregates from Ophiocordycipitaceae. Fungal Diversity 120(1): 1–76. 10.1007/s13225-023-00517-4 [DOI] [Google Scholar]
Xiao YP, Wen TC, Hongsanan S, et al. (2018). Multigene phylogenetics of Polycephalomyces (Ophiocordycipitaceae, Hypocreales), with two new species from Thailand. Scientific Reports 8: 18087. 10.1038/s41598-018-36792-4 [DOI] [PMC free article] [PubMed] [Google Scholar]
Xiao YP, Yang Y, Jayawardena RS, et al. (2024). Four novel Pleurocordyceps (Polycephalomycetaceae) species from China. Frontiers in Microbiology 14: 1256967. 10.3389/fmicb.2023.1256967 [DOI] [PMC free article] [PubMed] [Google Scholar]
Xie SW, Wei DP, Qiu JZ, et al. (2025). Overview of hirsutella-like anamorphs in Ophiocordyceps: introducing two new species and one new record from China. Mycokeys 119: 95–121. 10.3897/mycokeys.119.145174 [DOI] [PMC free article] [PubMed] [Google Scholar]
Yapa AT, Thambugala KM, Samarakoon MC, et al. (2024). Metarhizium species as bioinsecticides: potential, progress, applications & future perspectives. New Zealand Journal of Botany 2024: 1–23. 10.1080/0028825X.2024.2325006 [DOI] [Google Scholar]
Yu FM, Jayawardena RS, Luangharn T, et al. (2024). Species diversity of fungal pathogens on cultivated mushrooms: a case study on morels (Morchella, Pezizales). Fungal Diversity 5: 1–64. 10.1007/s13225-023-00531-6 [DOI] [Google Scholar]
Yu FM, Wei DP, Zhao Q, et al. (2023). Ascopolyporus tibetensis (Cordycipitaceae, Hypocreales): a new species from Tibet, China. Phytotaxa 592(2): 88–98. 10.11646/phytotaxa.592.2.2 [DOI] [Google Scholar]
Yu LQ, Zou X, Zhang J. (2017). A new Hirsutella species parasitic on grub larvae (Coleoptera). Microbiology China 44(6): 1370–1376. 10.13344/j.microbiol.china.160617 [DOI] [Google Scholar]
Zare R, Gams W. (2016). More white verticillium-like anamorphs with erect conidiophores. Mycological Progress 15(10): 993–1030. 10.1007/s11557-016-1214-8 [DOI] [Google Scholar]
Zeng XY, Tan TJ, Tian FH, et al. (2023). OFPT: a one-stop software for fungal phylogeny. Mycosphere 14(1): 1730–1741. 10.5943/mycosphere/14/1/20 [DOI] [Google Scholar]
Zhang D, Gao F, Jakovlić I, et al. (2020). PhyloSuite: An integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies. Molecular Ecology Resources 20(1): 348–355. 10.1111/1755-0998.13096 [DOI] [PubMed] [Google Scholar]
Zhang ZF, Zhou SY, Eurwilaichitr L, et al. (2021). Culturable mycobiota from Karst caves in China II, with descriptions of 33 new species. Fungal Diversity 106(1): 29–136. 10.1007/s13225-020-00453-7 [DOI] [Google Scholar]
Zhang ZY, Feng Y, Tong SQ, et al. (2023). Morphological and phylogenetic characterisation of two new soil-borne fungal taxa belonging to Clavicipitaceae (Hypocreales, Ascomycota). MycoKeys 98: 113–132. 10.3897/mycokeys.98.106240 [DOI] [PMC free article] [PubMed] [Google Scholar]
Zhu JY, Tang DX, Liu ZH, et al. (2023). Calcarisporium yuanyangense sp. nov. (Calcarisporiaceae, Hypocreales), a fungal pathogen isolated from Ophiocordyceps nutans. Phytotaxa 612(3): 27–282. 10.11646/phytotaxa.612.3.3 [DOI] [Google Scholar]
Zhuang Y, Luo C, Tang D, et al. (2025). Cretaceous entomopathogenic fungi illuminate the early evolution of insect-fungal associations. Proceedings of the Royal Society B: Biological Sciences 292(2048): 20250407. 10.1098/rspb.2025.0407 [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Table S1

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S2

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S3

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S4

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

Table S5

370TableS1toS5.xlsx^{(89.5KB, xlsx)}

[R1] Alves VC, Lira RA, Lima JM, et al. (2022). Unravelling the fungal darkness in a tropical cave: richness and the description of one new genus and six new species. Fungal Systematics and Evolution 10(1): 139–167. 10.3114/fuse.2022.10.06 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R2] Andersen SB, Ferrari M, Evans HC, et al. (2012). Disease dynamics in a specialized parasite of ant societies. PloS ONE 7(5): e36352. 10.1371/journal.pone.0036352 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R3] Aneja M, Gianfagna TJ, Hebbar PK. (2005). Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens. Physiological and Molecular Plant Pathology 67(6): 304–307. 10.1016/j.pmpp.2006.05.002 [DOI] [Google Scholar]

[R4] Araújo JPM, Lebert BM, Vermeulen S, et al. (2022). Masters of the manipulator: two new hypocrealean genera, Niveomyces (Cordycipitaceae) and Torrubiellomyces (Ophiocordycipitaceae), parasitic on the zombie ant fungus Ophiocordyceps camponoti-floridani. Persoonia 49(1): 171–194. 10.3767/persoonia.2022.49.05 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R5] Ballesteros-Aguirre CE, Sanjuan T, Ramírez-Cruz V, et al. (2025). Zombie-ant fungi from western Mexico: six new species in the Ophiocordyceps unilateralis complex (Hypocreales: Ascomycota) and a new host association with Cephalotes ants. Persoonia 55(1): 203–238. 10.3114/persoonia.2025.55.06 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R6] Ban S, Sakane T, Toyama K, et al. (2009). Teleomorph–anamorph relationships and reclassification of Cordyceps cuboidea and its allied species. Mycoscience 50(4): 261–272. 10.1007/S10267-008-0480-Y [DOI] [Google Scholar]

[R7] Bandelt HJ, Dress AWM. (1992). Split decomposition: a new and useful approach to phylogenetic analysis of distance data. Molecular Phylogenetics and Evolution 1(3): 242–252. 10.1016/1055-7903(92)90021-8 [DOI] [PubMed] [Google Scholar]

[R8] Bao DF, Hyde KD, Maharachchikumbura SS, et al. (2023). Taxonomy, phylogeny and evolution of freshwater Hypocreomycetidae (Sordariomycetes). Fungal Diversity 121(1): 1–94. 10.1007/s13225-023-00521-8 [DOI] [Google Scholar]

[R9] Berbee ML, Taylor JW. (2010). Dating the molecular clock in fungi – how close are we? Fungal Biology Reviews 24(1–2): 1–16. 10.1016/j.fbr.2010.03.001 [DOI] [Google Scholar]

[R10] Bermúdez-Cova MA, Haelewaters D, de Bekker C, et al. (2023). Hyperparasitic fungi—definitions, diversity, ecology, and research. Authorea. 10.22541/au.168787020.07281183/v2 [DOI] [Google Scholar]

[R11] Bischoff JF, Chaverri P, White JF., Jr (2005). Clarification of the host substrate of Ascopolyporus and description of Ascopolyporus philodendrus sp. nov. Mycologia 97(3): 710–717. 10.3852/mycologia.97.3.710 [DOI] [PubMed] [Google Scholar]

[R12] Candoussau F, Boqueras M, Gómez-Bolea A, et al. (2007). Observations on Neobarya, including new species and new combinations. Sydowia 59(2): 179–215. [Google Scholar]

[R13] Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T. (2009). trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15): 1972–1973. 10.1093/bioinformatics/btp348 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R14] Chaverri P, Liu M, Hodge KT. (2008). A monograph of the entomopathogenic genera Hypocrella, Moelleriella, and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their aschersonia-like anamorphs in the Neotropics. Studies in Mycology 60: 1–66. 10.3114/sim.2008.60.01 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R15] Chen WH, Han YF, Liang JD, et al. (2017). Metarhizium dendrolimatilis, a novel Metarhizium species parasitic on Dendrolimus sp. larvae. Mycosphere 8: 31–37. 10.5943/mycosphere/8/1/4 [DOI] [Google Scholar]

[R16] Chen WH, Han YF, Liang JD, et al. (2019). Morphological and phylogenetic characterization of novel Metarhizium species in Guizhou, China. Phytotaxa 419(2): 189–196. 10.11646/phytotaxa.419.2.5 [DOI] [Google Scholar]

[R17] Chen WH, Li D, Liang J D, et al. (2024b). Two new Cordyceps-like species, Perennicordyceps zongqii sp. nov. (Polycephalomycetaceae) and Purpureocillium zongqii sp. nov. (Ophiocordycipitaceae), in Hypocreales from karst region of China. MycoKeys 110: 141–158. 10.3897/mycokeys.110.135724 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R18] Chen WH, Li D, Liang JD, et al. (2024a). Chlorocillium sinense sp. nov. (Clavicipitaceae) and Calcarisporium guizhouense sp. nov. (Calcarisporiaceae) in Hypocreales from China. MycoKeys 109: 91–107. 10.3897/mycokeys.109.128060 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R19] Chen WH, Li D, Shu HL, et al. (2025a). Four new araneogenous species and a new genus in Hypocreales (Clavicipitaceae, Cordycipitaceae) from the karst region of China. MycoKeys 112: 335–359. 10.3897/mycokeys.112.140799 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R20] Chen WH, Liang JD, Ren XX, et al. (2022). Phylogenetic, ecological and morphological characteristics reveal two new spider-associated genera in Clavicipitaceae. MycoKeys 91: 49–66. 10.3897/mycokeys.91.86812 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R21] Chen WH, Shu HL, Li D, et al. (2025b). Shedding light on the darkness: Cryptic diversity of cordyceps-like fungi in karst regions of Guizhou Province, China. Mycosphere 16: 2887–2974. 10.5943/mycosphere/16/1/20 [DOI] [Google Scholar]

[R22] Chernomor O, Von Haeseler A, Minh BQ. (2016). Terrace aware data structure for phylogenomic inference from supermatrices. Systematic Biology 65(6): 997–1008. 10.1093/sysbio/syw037 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R23] Clewley JP. (1995). Macintosh sequence analysis software. Molecular Biotechnology 3: 221–224. 10.1007/BF02789332 [DOI] [PubMed] [Google Scholar]

[R24] Crous PW, Costa MM, Kandemir H, et al. (2023). Fungal Planet description sheets: 1550–1613. Persoonia 51: 280–417. 10.3767/persoonia.2023.51.08 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R25] Crous PW, Gams W, Stalpers JA, et al. (2004). MycoBank: an online initiative to launch mycology into the 21st century. Studies in Mycology 50: 19–22. [Google Scholar]

[R26] Custódio FA, Pereira OL. (2025). New treasures in Cordycipitaceae: fungicolous fungi associated with Pseudocercospora fijiensis and P. musae in Brazil, including Matutinistella gen. nov. Fungal Systematics and Evolution 15(1): 133–152. 10.3114/fuse.2025.15.06 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R27] Dai Y, Chen S, Wang Y, et al. (2024). Molecular phylogenetics of the Ophiocordyceps sinensis-species complex lineage (Ascomycota, Hypocreales), with the discovery of new species and predictions of species distribution. IMA Fungus 15(1): 2. 10.1186/s43008-023-00131-8 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R28] Darmostuk V, Etayo J, Rodriguez-Flakus P, et al. (2025). A novel, exclusively lichen-inhabiting lineage of hypocrealean fungi revealed in the Sordariomycetes. Persoonia 54: 47–91. 10.3114/persoonia.2025.54.02 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R29] Ding M, Li Y, Bai Y, et al. (2024). Albacillium hingganense gen. et sp. nov. (Clavicipitaceae) from forest litters in Northeast China. Phytotaxa 650(2): 157–168. 10.11646/phytotaxa.650.2.3 [DOI] [Google Scholar]

[R30] Dong W, Hyde KD, Jeewon R, et al. (2025). Fungal diversity notes 2017–2122: taxonomic and phylogenetic contributions to freshwater fungi and other fungal taxa. Fungal Diversity 134: 185–459. 10.1007/s13225-025-00560-3 [DOI] [Google Scholar]

[R31] El-Hasan A, Walker F, Klaiber I, et al. (2022). New approaches to manage Asian soybean rust (Phakopsora pachyrhizi) using Trichoderma spp. or their antifungal secondary metabolites. Metabolites 12(6): 507. 10.3390/metabo12060507 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R32] Flakus A, Etayo J, Miadlikowska J, et al. (2019). Biodiversity assessment of ascomycetes inhabiting Lobariella lichens in Andean cloud forests led to one new family, three new genera and 13 new species of lichenicolous fungi. Plant and Fungal Systematics 64(2): 283–344. 10.2478/pfs-2019-0022 [DOI] [Google Scholar]

[R33] Gams W. (1971). Cephalosporium-artige Schimmelpilze (Hyphomycetes). Gustav Fischer Verlag, Stuttgart. [Google Scholar]

[R34] Gonzalez MC, Glenn AE, Hanlin RT, et al. (2016). Acremonium camptosporum isolated as an endophyte of Bursera simaruba from Yucatan Peninsula, Mexico. Mycotaxon 131(1): 211–225. [Google Scholar]

[R35] Guindon S, Dufayard JF, Lefort V, et al. (2010). New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic Biology 59(3): 307–321. 10.1093/sysbio/syq010 [DOI] [PubMed] [Google Scholar]

[R36] Hall TA. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98. [Google Scholar]

[R37] Hasan R, Lv B, Uddin MJ, et al. (2022). Monitoring mycoparasitism of Clonostachys rosea against Botrytis cinerea using GFP. Journal of Fungi 8(6): 567. 10.3390/jof8060567 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R38] Hennings P. (1902). Fungi blumenaevienses II. a cl. Alfr. Möller lecti. Hedwigia 41(1): 1–33. [Google Scholar]

[R39] Hennings P. (1909). Fungi S. Paulensis IV a cl. Puttemans collecti. Hedwigia 48: 1–20. [Google Scholar]

[R40] Hongsanan S, Maharachchikumbura SSN, Hyde KD, et al. (2017). An updated phylogeny of Sordariomycetes based on phylogenetic and molecular clock evidence. Fungal Diversity 84(1): 25–41. 10.1007/s13225-017-0384-2 [DOI] [Google Scholar]

[R41] Hoog GS. (1978). Notes on some fungicolous hyphomycetes and their relatives. Persoonia 10(1): 33–81. [Google Scholar]

[R42] Hou LW, Giraldo A, Groenewald JZ, et al. (2023). Redisposition of acremonium-like fungi in Hypocreales. Studies in Mycology 105: 23–203. 10.3114/sim.2023.105.02 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R43] Huson DH, Bryant D. (2024). The SplitsTree app: interactive analysis and visualization using phylogenetic trees and networks. Nature Methods 21(10): 1773–1774. 10.1038/s41592-024-02406-3 [DOI] [PubMed] [Google Scholar]

[R44] Hyde KD, Noorabadi M T, Thiyagaraja V, et al. (2024). The 2024 outline of Fungi and fungus-like taxa. Mycosphere 15(1): 5146–6239. 10.5943/mycosphere/15/1/25 [DOI] [Google Scholar]

[R45] Hyde KD, Maharachchikumbura SS, Hongsanan S, et al. (2017). The ranking of fungi: a tribute to David L. Hawksworth on his 70th birthday. Fungal Diversity 84: 1–23. 10.1007/s13225-017-0383-3 [DOI] [Google Scholar]

[R46] Johnston PR, Park D. (2023). Dingleyomyces lloydii (Hypocreales, Polycephalomycetaceae), gen. nov., comb. nov. New Zealand Journal of Botany 63(4): 851–858. 10.1080/0028825X.2023.2291453 [DOI] [Google Scholar]

[R47] Kaitsu Y, Shimizu K, Tanaka E, et al. (2013). Ophiocordyceps sessilis sp. nov., a new species of Ophiocordyceps on Camponotus ants in Japan. Mycological Progress 12: 755–761. 10.1007/s11557-013-0911-9 [DOI] [Google Scholar]

[R48] Kalyaanamoorthy S, Minh BQ, Wong TK, et al. (2017). ModelFinder: fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. 10.1038/nmeth.4285 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R49] Katoh K, Rozewicki J, Yamada KD. (2019). MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20: 1160–1166. 10.1093/bib/bbx108 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R50] Kepler RM, Sung GH, Harada Y, et al. (2012). Host jumping onto close relatives and across kingdoms by Tyrannicordyceps (Clavicipitaceae) gen. nov. and Ustilaginoidea (Clavicipitaceae). American Journal of Botany 99(3): 552–561. 10.3732/ajb.1100124 [DOI] [PubMed] [Google Scholar]

[R51] Kepler RM, Luangsa-Ard JJ, Hywel-Jones NL, et al. (2017). A phylogenetically-based nomenclature for Cordycipitaceae (Hypocreales). IMA Fungus 8(2): 335–353. 10.5598/imafungus.2017.08.02.08 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R52] Khao-Ngam S, Mongkolsamrit S, Rungjindamai N, et al. (2021). Ophiocordyceps asiana and Ophiocordyceps tessaratomidarum (Ophiocordycipitaceae, Hypocreales), two new species on stink bugs from Thailand. Mycological Progress 20: 341–353. 10.1007/s11557-021-01684-x [DOI] [Google Scholar]

[R53] Khonsanit A, Luangsa-Ard JJ, Thanakitpipattana D, et al. (2019). Cryptic species within Ophiocordyceps myrmecophila complex on formicine ants from Thailand. Mycological Progress 18: 147–161. 10.1007/s11557-018-1412-7 [DOI] [Google Scholar]

[R54] Khonsanit A, Thanakitpipattana D, Mongkolsamrit S, et al. (2024). A phylogenetic assessment of Akanthomyces sensu lato in Cordycipitaceae (Hypocreales, Sordariomycetes): introduction of new genera, and the resurrection of Lecanicillium. Fungal Systematics and Evolution 14(1): 271–306. 10.3114/fuse.2024.14.17 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R55] Kobmoo N, Mongkolsamrit S, Khonsanit A, et al. (2024). Integrative taxonomy of Metarhizium anisopliae species complex, based on phylogenomics combined with morphometrics, metabolomics, and virulence data. IMA Fungus 15(1): 30. 10.1186/s43008-024-00154-9 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R56] Kobmoo N, Tasanathai K, Araújo JP, et al. (2023). New mycoparasitic species in the genera Niveomyces and Pseudoniveomyces gen. nov. (Hypocreales: Cordycipitaceae), with sporothrix-like asexual morphs, from Thailand. Fungal Systematics and Evolution 12(1): 91–110. 10.3114/fuse.2023.12.07 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R57] Kornerup A, Wanscher JH. (1978). Methuen Handbook of Colour, 3rd edn. Eyre Methuen Ltd., London. [Google Scholar]

[R58] Kryukov V, Yaroslavtseva O, Tyurin M, et al. (2017). Ecological preferences of Metarhizium spp. from Russia and neighboring territories and their activity against Colorado potato beetle larvae. Journal of Invertebrate Pathology 149: 1–7. 10.1016/j.jip.2017.07.001 [DOI] [PubMed] [Google Scholar]

[R59] Kuraku S, Zmasek CM, Nishimura O, et al. (2013). aLeaves facilitates on-demand exploration of metazoan gene family trees on MAFFT sequence alignment server with enhanced interactivity. Nucleic Acids Research 41: W22–W28. 10.1093/nar/gkt389 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R60] Larsson A. (2014). AliView: a fast and lightweight alignment viewer and editor for large datasets. Bioinformatics 30(22): 3276–3278. 10.1093/bioinformatics/btu531 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R61] Lee K, Missaoui A, Mahmud K, et al. (2021). Interaction between grasses and Epichloë endophytes and its significance to biotic and abiotic stress tolerance and the rhizosphere. Microorganisms 9(11): 2186. 10.3390/microorganisms9112186 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R62] Lin L, Lin Y, Keyhani NO, et al. (2025). New entomopathogenic species in the Clavicipitaceae family (Hypocreales, Ascomycota) from the subtropical forests of Fujian, China. Frontiers in Microbiology 16: 1532341. 10.3389/fmicb.2025.1532341 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R63] Liu M, Shoukouhi P, Bisson K R, et al. (2021b). Sympatric divergence of the ergot fungus, Claviceps purpurea, populations infecting agricultural and nonagricultural grasses in North America. Ecology and Evolution 11(1): 273–293. 10.1002/ece3.7028. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R64] Liu Q, Dong C. (2023). Dual transcriptomics reveals interspecific interactions between the mycoparasite Calcarisporium cordycipiticola and its host Cordyceps militaris. Microbiology Spectrum 11(2): e04800-22. 10.1128/spectrum.04800-22 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R65] Liu Q, Wang F, Xu FX, et al. (2021a). Pathogenic fungi of artificially cultivated Cordyceps militaris. Mycosystema 40(11): 2962–2980. 10.13346/j.mycosystema.210147 [DOI] [Google Scholar]

[R66] Liu Z, Tang D, Lu Y, et al. (2024). Morphology and phylogeny of four new species within Polycephalomycetaceae (Hypocreales) parasitising Ophiocordyceps species. MycoKeys 105: 179–202. 10.3897/mycokeys.105.119893 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R67] Ma JM, Wang ZQ, Yang ZL, et al. (2024). Metarhizium puerense (Hypocreales, Clavicipitaceae): a new species from Yunnan, south-western China. Biodiversity Data Journal 12: e129087. 10.3897/BDJ.12.e129087 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R68] Maharachchikumbura SSN, Hyde KD, Jones EBG, et al. (2016). Families of Sordariomycetes. Fungal Diversity 79: 1–317. 10.1007/s13225-016-0369-6 [DOI] [Google Scholar]

[R69] Matsushima T. (1975). Icones Microfungorum a Matsushima Lectorum. Published by the author. Kobe, Japan. [Google Scholar]

[R70] Minh BQ, Nguyen MA, Von Haeseler A. (2013). Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution 30(5): 1188–1195. 10.1093/molbev/mst024 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R71] Möller A. (1901). Phycomyceten und Ascomyceten. Untersuchungen aus Brasilien. Botanische Mittheilungen aus den Tropen 9: 1–319. 10.5962/bhl.title.2753 [DOI] [Google Scholar]

[R72] Mongkolsamrit S, Khonsanit A, Thanakitpipattana D, et al. (2020). Revisiting Metarhizium and the description of new species from Thailand. Studies in Mycology 95(1): 171–251. 10.1016/j.simyco.2020.04.001 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R73] Mongkolsamrit S, Noisripoom W, Tasanathai K, et al. (2022). Comprehensive treatise of Hevansia and three new genera Jenniferia, Parahevansia and Polystromomyces on spiders in Cordycipitaceae from Thailand. MycoKeys 91: 113–145. 10.3897/mycokeys.91.83091 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R74] Mongkolsamrit S, Noisripoom W, Thanakitpipattana D, et al. (2018). Disentangling cryptic species with Isaria-like morphs in Cordycipitaceae. Mycologia 110(1): 230–257. 10.1080/00275514.2018.1446651 [DOI] [PubMed] [Google Scholar]

[R75] Mongkolsamrit S, Sandargo B, Ebada SS, et al. (2023). Bhushaniella gen. nov. (Cordycipitaceae) on spider eggs sac: a new genus from Thailand and its bioactive secondary metabolites. Mycological Progress 22(9): 64. 10.1007/s11557-023-01915-3 [DOI] [Google Scholar]

[R76] Mukherjee PK, Mendoza-Mendoza A, Zeilinger S, et al. (2022). Mycoparasitism as a mechanism of Trichoderma-mediated suppression of plant diseases. Fungal Biology Reviews 39: 15–33. 10.1016/j.fbr.2021.11.004 [DOI] [Google Scholar]

[R77] Nguyen LT, Schmidt HA, Von Haeseler A, et al. (2015). IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268–274. 10.1093/molbev/msu300 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R78] Nguyen TT, Le TN, Nguyen TH. (2023). First report of emerging fungal pathogens of Cordyceps militaris in Vietnam. Scientific Reports 13(1): 17669. 10.1038/s41598-023-43951-9 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R79] Nylander JAA. (2004). MrModeltest 2.2: program distributed by the author. Evolutionary Biology Centre, Uppsala University. [Google Scholar]

[R80] Peng XC, Wen TC, Wei DP, et al. (2024). Two new species and one new combination of Ophiocordyceps (Hypocreales, Ophiocordycipitaceae) in Guizhou. MycoKeys 102: 245–266. 10.3897/mycokeys.102.113351 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R81] Petch T. (1926). Entomogenous fungi: additions and corrections. II. Transactions of the British Mycological Society 11(3–4): 258–266. 10.1016/S0007-1536(26)80010-5 [DOI] [Google Scholar]

[R82] Preuss CGT. (1851). Synopsis fungorum hucusque cognitorum praesertim prope Hoyerswerda. Linnaea 24: 101–153. [Google Scholar]

[R83] Quandt CA, Kepler RM, Gams W, et al. (2014). Phylogenetic-based nomenclatural proposals for Ophiocordycipitaceae (Hypocreales) with new combinations in Tolypocladium. IMA Fungus 5: 121–134. 10.5598/imafungus.2014.05.01.12 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R84] Rambaut A. (2018). FigTree v. 1.4.4. Available online: http://tree.bio.ed.ac.uk/software/figtree/.

[R85] Rambaut A, Suchard MA, Xie D, et al. (2013). Tracer version 1.6. University of Edinburgh. [Google Scholar]

[R86] Ronquist F, Teslenko M, van der Mark P, et al. (2012). MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systems Biology 61: 539–542. 10.1093/sysbio/sys029 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R87] Samarakoon MC, Hyde KD, Maharachchikumbura SSM, et al. (2022). Taxonomy, phylogeny, molecular dating and ancestral state reconstruction of Xylariomycetidae (Sordariomycetes). Fungal Diversity 112(1): 1–88. 10.1007/s13225-021-00495-5 [DOI] [Google Scholar]

[R88] Sawa R, Mori Y, Iinuma H, et al. (1994). Harzianic acid, a new antimicrobial antibiotic from a fungus. The Journal of Antibiotics 47(6): 731–732. 10.7164/antibiotics.47.731 [DOI] [PubMed] [Google Scholar]

[R89] Shen J, Gou W, Tian HL, et al. (2026). Molecular phylogeny and morphology reveal two new species, Ophiocordyceps ramosa sp. nov. (Ophiocordycipitaceae) and Leptobacillium hepiali sp. nov. (Cordycipitaceae), in Hypocreales from China. MycoKeys 127: 191. 10.3897/mycokeys.127.173361 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R90] Somrithipol S, Jones EBG. (2006). Calcarisporium phaeopodium sp. nov., a new hyphomycete from Thailand. Sydowia 58(1): 133–140. [Google Scholar]

[R91] Steel MA. (1994). Recovering a tree from the leaf colourations it generates under a Markov model. Applied Mathematics Letters 7(2): 19–24. 10.1016/0893-9659(94)90024-8 [DOI] [Google Scholar]

[R92] Subramanian CV. (1978). Revisions of Hyphomycetes I. Kavaka 5: 93–98. [Google Scholar]

[R93] Sun JZ, Dong CH, Liu XZ, et al. (2016). Calcarisporium cordycipiticola sp. nov., an important fungal pathogen of Cordyceps militaris. Phytotaxa 268(2): 135–144. 10.11646/phytotaxa.268.2.4 [DOI] [Google Scholar]

[R94] Sun JZ, Liu XZ, Hyde KD, et al. (2017). Calcarisporium xylariicola sp. nov. and introduction of Calcarisporiaceae fam. nov. in Hypocreales. Mycological Progress 16(4): 433–445. 10.1007/s11557-017-1290-4 [DOI] [Google Scholar]

[R95] Sun JZ, Liu XZ, McKenzie EH, et al. (2019). Fungicolous fungi: terminology, diversity, distribution, evolution, and species checklist. Fungal Diversity 95: 337–430. 10.1007/s13225-019-00422-9 [DOI] [Google Scholar]

[R96] Sung GH, Hywel-Jones NL, Sung JM, et al. (2007). Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Studies in Mycology 57: 5–59. 10.3114/sim.2007.57.01 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R97] Sung GH, Poinar GO, Jr, Spatafora JW. (2008). The oldest fossil evidence of animal parasitism by fungi supports a Cretaceous diversification of fungal–arthropod symbioses. Molecular Phylogenetics and Evolution 49: 495–502. 10.1016/j.ympev.2008.08.028 [DOI] [PubMed] [Google Scholar]

[R98] Tan TP, Valter S, Lacey E. (2025). Nomenclatural novelties. Index of Australian Fungi 62: 1–20. 10.5281/zenodo.17336362 [DOI] [Google Scholar]

[R99] Tan YP, Minns SA, Valter S, et al. (2024). Nomenclatural novelties. Index of Australian Fungi 47: 1–14. 10.5281/zenodo.14177440 [DOI] [Google Scholar]

[R100] Tan YP, Shivas RG. (2023a). Nomenclatural novelties. Index of Australian Fungi 22: 1–14. 10.5281/zenodo.10145676 [DOI] [Google Scholar]

[R101] Tan YP, Shivas RG. (2023b). Nomenclatural novelties. Index of Australian Fungi 27: 1–18. 10.5281/zenodo.10426579 [DOI] [Google Scholar]

[R102] Tan YP, Shivas RG. (2024b). Nomenclatural novelties. Index of Australian Fungi 31: 1–12. 10.5281/zenodo.10784071 [DOI] [Google Scholar]

[R103] Tasanathai K, Thanakitpipattana D, Himaman W, et al. (2020). Three new Ophiocordyceps species in the Ophiocordyceps pseudoacicularis species complex on Lepidoptera larvae in Southeast Asia. Mycological Progress 19(10): 1043–1056. 10.1007/s11557-020-01611-6 [DOI] [Google Scholar]

[R104] Thanakitpipattana D, Mongkolsamrit S, Khonsanit A, et al. (2022). Is Hyperdermium congeneric with Ascopolyporus? Phylogenetic relationships of Ascopolyporus spp. (Cordycipitaceae, Hypocreales) and a new genus Neohyperdermium on scale insects in Thailand. Journal of Fungi 8(5): 516. 10.3390/jof8050516 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R105] Vaidya G, Lohman DJ, Meier R. (2011). SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27: 171–180. 10.1111/j.1096-0031.2010.00329.x [DOI] [PubMed] [Google Scholar]

[R106] Wang L, Li HH, Chen YQ, et al. (2014). Polycephalomyces lianzhouensis sp. nov., a new species, co-occurs with Ophiocordyceps crinalis. Mycological Progress 13: 1089–1096. 10.1007/s11557-014-0996-9 [DOI] [Google Scholar]

[R107] Wang Y, Wei D P, Peng X C, et al. (2024). Interesting mycoparasites and Paradingleyomyces lepidopterorum gen. et sp. nov. (Hypocreales, Polycephalomycetaceae) from Yunnan Province, China. MycoKeys 110: 185–210. 10.3897/mycokeys.110.134132 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R108] Wang YB, Wang Y, Fan Q, et al. (2020). Multigene phylogeny of the family Cordycipitaceae (Hypocreales): new taxa and the new systematic position of the Chinese cordycipitoid fungus Paecilomyces hepiali. Fungal Diversity 103: 1–46. 10.1007/s13225-020-00457-3 [DOI] [Google Scholar]

[R109] Wang YH, Ban S, Wang WJ, et al. (2021). Pleurocordyceps gen. nov. for a clade of fungi previously included in Polycephalomyces based on molecular phylogeny and morphology. Journal of Systematics and Evolution 59(5): 1065–1080. 10.1111/jse.12705 [DOI] [Google Scholar]

[R110] Wei DP, Gentekaki E, Wanasinghe DN, et al. (2022). Diversity, molecular dating and ancestral characters state reconstruction of entomopathogenic fungi in Hypocreales. Mycosphere 13(2): 281–351. 10.5943/mycosphere/si/1f/8 [DOI] [Google Scholar]

[R111] Wei DP, Wanasinghe DN, Hyde KD, et al. (2019). The genus Simplicillium. MycoKeys 60: 69. 10.3897/mycokeys.60.38040 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R112] Wei DP, Wanasinghe DN, Hyde KD, et al. (2020). Ophiocordyceps tianshanensis sp. nov. on ants from Tianshan mountains, P. R. China. Phytotaxa 464(4): 277–292. 10.11646/phytotaxa.464.4.2 [DOI] [Google Scholar]

[R113] Wei DP, Wanasinghe DN, Xu JC, et al. (2021). Three novel entomopathogenic fungi from China and Thailand. Frontiers in Microbiology 11: 608991. 10.3389/fmicb.2020.608991 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R114] Wen TC, Xiao YP, Han YF, et al. (2017). Multigene phylogeny and morphology reveal that the Chinese medicinal mushroom ‘Cordyceps gunnii’ is Metacordyceps neogunnii sp. nov. Phytotaxa 302(1): 27–39. 10.11646/phytotaxa.302.1.2 [DOI] [Google Scholar]

[R115] Wijayawardene NN, Hyde KD, Al-Ani LKT, et al. (2020). Outline of Fungi and fungus-like taxa. Mycosphere 11: 1060–1456. 10.5943/mycosphere/11/1/8 [DOI] [Google Scholar]

[R116] Wijayawardene NN, Hyde KD, Dai DQ, et al. (2022). Outline of Fungi and fungus-like taxa – 2021. Mycosphere 13(1): 53–453. 10.5943/mycosphere/13/1/2 [DOI] [Google Scholar]

[R117] Wijayawardene NN, Hyde KD, Lumbsch HT, et al. (2018). Outline of Ascomycota: 2017. Fungal Diversity 88: 167–263. 10.1007/s13225-018-0394-8 [DOI] [Google Scholar]

[R118] Xiang CY, Gao F, Jakovlić I. et al. (2023). Using PhyloSuite for molecular phylogeny and tree‐based analyses. iMeta 1: e87. 10.1002/imt2.87 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R119] Xiao YP, Hongsanan S, Hyde KD, et al. (2019). Two new entomopathogenic species of Ophiocordyceps in Thailand. MycoKeys 47: 53–74. 10.3897/mycokeys.47.29898 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R120] Xiao YP, Wang YB, Hyde KD, et al. (2023). Polycephalomycetaceae, a new family of clavicipitoid fungi segregates from Ophiocordycipitaceae. Fungal Diversity 120(1): 1–76. 10.1007/s13225-023-00517-4 [DOI] [Google Scholar]

[R121] Xiao YP, Wen TC, Hongsanan S, et al. (2018). Multigene phylogenetics of Polycephalomyces (Ophiocordycipitaceae, Hypocreales), with two new species from Thailand. Scientific Reports 8: 18087. 10.1038/s41598-018-36792-4 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R122] Xiao YP, Yang Y, Jayawardena RS, et al. (2024). Four novel Pleurocordyceps (Polycephalomycetaceae) species from China. Frontiers in Microbiology 14: 1256967. 10.3389/fmicb.2023.1256967 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R123] Xie SW, Wei DP, Qiu JZ, et al. (2025). Overview of hirsutella-like anamorphs in Ophiocordyceps: introducing two new species and one new record from China. Mycokeys 119: 95–121. 10.3897/mycokeys.119.145174 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R124] Yapa AT, Thambugala KM, Samarakoon MC, et al. (2024). Metarhizium species as bioinsecticides: potential, progress, applications & future perspectives. New Zealand Journal of Botany 2024: 1–23. 10.1080/0028825X.2024.2325006 [DOI] [Google Scholar]

[R125] Yu FM, Jayawardena RS, Luangharn T, et al. (2024). Species diversity of fungal pathogens on cultivated mushrooms: a case study on morels (Morchella, Pezizales). Fungal Diversity 5: 1–64. 10.1007/s13225-023-00531-6 [DOI] [Google Scholar]

[R126] Yu FM, Wei DP, Zhao Q, et al. (2023). Ascopolyporus tibetensis (Cordycipitaceae, Hypocreales): a new species from Tibet, China. Phytotaxa 592(2): 88–98. 10.11646/phytotaxa.592.2.2 [DOI] [Google Scholar]

[R127] Yu LQ, Zou X, Zhang J. (2017). A new Hirsutella species parasitic on grub larvae (Coleoptera). Microbiology China 44(6): 1370–1376. 10.13344/j.microbiol.china.160617 [DOI] [Google Scholar]

[R128] Zare R, Gams W. (2016). More white verticillium-like anamorphs with erect conidiophores. Mycological Progress 15(10): 993–1030. 10.1007/s11557-016-1214-8 [DOI] [Google Scholar]

[R129] Zeng XY, Tan TJ, Tian FH, et al. (2023). OFPT: a one-stop software for fungal phylogeny. Mycosphere 14(1): 1730–1741. 10.5943/mycosphere/14/1/20 [DOI] [Google Scholar]

[R130] Zhang D, Gao F, Jakovlić I, et al. (2020). PhyloSuite: An integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies. Molecular Ecology Resources 20(1): 348–355. 10.1111/1755-0998.13096 [DOI] [PubMed] [Google Scholar]

[R131] Zhang ZF, Zhou SY, Eurwilaichitr L, et al. (2021). Culturable mycobiota from Karst caves in China II, with descriptions of 33 new species. Fungal Diversity 106(1): 29–136. 10.1007/s13225-020-00453-7 [DOI] [Google Scholar]

[R132] Zhang ZY, Feng Y, Tong SQ, et al. (2023). Morphological and phylogenetic characterisation of two new soil-borne fungal taxa belonging to Clavicipitaceae (Hypocreales, Ascomycota). MycoKeys 98: 113–132. 10.3897/mycokeys.98.106240 [DOI] [PMC free article] [PubMed] [Google Scholar]

[R133] Zhu JY, Tang DX, Liu ZH, et al. (2023). Calcarisporium yuanyangense sp. nov. (Calcarisporiaceae, Hypocreales), a fungal pathogen isolated from Ophiocordyceps nutans. Phytotaxa 612(3): 27–282. 10.11646/phytotaxa.612.3.3 [DOI] [Google Scholar]

[R134] Zhuang Y, Luo C, Tang D, et al. (2025). Cretaceous entomopathogenic fungi illuminate the early evolution of insect-fungal associations. Proceedings of the Royal Society B: Biological Sciences 292(2048): 20250407. 10.1098/rspb.2025.0407 [DOI] [PMC free article] [PubMed] [Google Scholar]

PERMALINK

Taxonomy, phylogeny and diversity of hyperparasites and their allied entomopathogens in Hypocreales

DP Wei

E Gentekaki

JJ Luangsa-ard

NN Wijayawardene

DN Wanasinghe

ZL Liu

QF Huang

NY Liu

SW Xie

JC Kang

TC Wen

Abstract

Introduction

Materials and methods

Sampling and isolation

Fig. 1.

Fig. 2.

Morphological observation

DNA extraction, PCR amplification and sequencing

Phylogenetic analyses

Genealogical concordance phylogenetic species recognition (GCPSR)

Divergence time estimation

Results

Phylogenetic analyses

Analyses 1 – The familial placements of Hypocreales

Fig. 3.

Fig. 4.

Fig. 5.

Analyses 2 - Interspecific relationship of Chlorocillium

Analyses 3 - Interspecific relationship of Pleurocordyceps

Analyses 4 - Interspecific relationship of Ophiocordyceps

Fig. 6.

Divergence time estimation

Fig. 7.

Table 1.

Taxonomy

Fig. 8.

Table 2.

Fig. 9.

Fig. 10.

Fig. 11.

Fig. 12.

Fig. 13.

Fig. 14.

Fig. 15.

Fig. 16.

Fig. 17.

Fig. 18.

Fig. 19.

Fig. 20.

Fig. 21.

Fig. 22.

Fig. 23.

Fig. 24.

Fig. 30.

Fig. 25.

Fig. 26.

Fig. 27.

Fig. 28.

Fig. 29.

Fig. 31.

Fig. 32.

Fig. 33.

Fig. 34.

Discussion

Establishment of Chlorocilliaceae and Ch. guizhouense complex

Biodiversity, host spectrum and trophic strategies of entomopathogens-associated hyperparasites in Hypocreales

Fig. 35.

Physiological impacts and ecological significance of hyperparasites

Fig. 36.

Acknowledgments

Footnotes

Supplementary Material: https://studiesinmycology.org/

References

Associated Data

Supplementary Materials

ACTIONS

PERMALINK