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. 2024 Oct 22;109:337–354. doi: 10.3897/mycokeys.109.133325

Three new wood-inhabiting fungi of Botryobasidium (Cantharellales, Basidiomycota) from subtropical forests of Southwestern China

Lin-Jiang Zhou 1,2, Xue-Long Li 3, Hai-Sheng Yuan 1,✉
PMCID: PMC11522739  PMID: 39478835

Abstract

The basidiomycete genus Botryobasidium is a resupinate saprotrophic with a global distribution range from coniferous to broad-leaved forest ecosystems. Though numerous species have been reported from Eurasia and North America, few have been described from China. In the current work, phylogenetic analyses of Botryobasidium in China were conducted based on the dataset of the internal transcribed spacer (ITS) regions and the large subunit (LSU) of nuclear ribosomal RNA gene. Maximum likelihood and Bayesian analyses were used to reconstruct the phylogenetic tree, and three new species, namely Botryobasidiumacanthosporum, B.leptocystidiatum and B.subovalibasidium, were described from subtropical forests of Yunnan Province, Southwestern China. Botryobasidiumacanthosporum is characterized by having yellowish white to dark yellow basidiome, clavate to tubular cystidia, and subglobose to globose basidiospores with obtuse spines. Botryobasidiumleptocystidiatum is characterized by having fluffy to arachnoid, greyish white to ivory basidiome, generative hyphae with clamped, tubular cystidia, and subnavicular to navicular basidiospores. While, B.subovalibasidium is characterized by having yellowish to ivory basidiome, subovoid basidia, navicular to suburniform basidiospores, and thick-walled chlamydospores. These three new species are described and illustrated, and the discriminating characters between the new species and their closely related species are discussed. A key to known species of Botryobasidium in China is provided.

Key words: Botryobasidiaceae, corticioid fungi, subtropical forests, taxonomy, wood-decaying fungi

Introduction

Botryobasidium Donk belongs to the order Cantharellales of phylum Basidiomycota, and was typified by B.subcoronatum (Höhn. & Litsch.) Donk (Moncalvo et al. 2006). Many asexual morph generic names, such as Acladium Link, Allescheriella Henn., Alysidium Kunze, Haplotrichum Link, Neoacladium P.N. Singh & S.K. Singh, Physospora Fr., and Sporocephalium Chevall., are congeneric with Botryobasidium, and were re-combined in Botryobasidium (Stalpers et al. 2021). The genus is characterized by resupinate, smooth, arachnoid, hypochnoid, pellicular or grandinioid basidiomes, a monomitic hyphal system with simple septate or nodose generative hyphae, clavate to cylindrical cystidia, claviform to suburniform basidia with 2–8 sterigmata, and navicular to globose, thin- or thick-walled, smooth or ornamented basidiospores (Binder et al. 2005; Larsson 2007; Buyck et al. 2017; Bondartseva and Zmitrovich 2023). In macromorphology, species of Botryobasidium are easily confused with some genera, e.g., Ceratobasidium D.P. Rogers, Sistotrema Fr., and Tulasnella J. Schröt. in the Cantharellales (Donk 1956; Oberwinkler 1982), whereas Botryobasidium differs from the others in absence of epibasidia, sturdy and long sterigmata, and oily inclusions (Kotiranta and Saarenoksa 2005; Gorjón and Hallenberg 2008; Oberwinkler et al. 2017). Subsequent molecular phylogenetic studies confirmed the close relationships among Botryobasidium, Cantharellus, Clavulina, Hydnum and Tulasnella (Jülich 1981; Hibbett et al. 1997; Bruns et al. 1998; Langer 1998; Pine et al. 1999; Cao et al. 2021).

The species of Botryobasidium are a group of saprotrophic fungi that cause a white rot in fallen angiosperm and gymnosperm woods, which play a key role in carbon recycling and energy flow in different forest ecosystems (Langer et al. 2000b; Bondartseva and Zmitrovich 2023). They can be commonly found on various hosts or substrates from the litter, fallen trunk to stem of living trees, including the macrophanerophytes, such as Abies Mill, Acer Linn., Alnus Mill., Betula L., Citrus L., Corylus L., Eucalyptus L. Herit, Fagus L., Magnolia Linn., Persea Mill., Picea Dietr., Pinus Linn, Populus L., Quercus L., Salix L. and Tsuga Carr.; the shrubs, such as Bambusa Retz. corr. Schreber and Pandanus Linn. f.; the pteridophytes, such as Pteris L. and Cibotium Kaulf. (Anon 1969; Holubová-Jechová 1969; Boidin and Gilles 1982; Langer 1994; Langer et al. 2000a, 2000b; Hjortstam et al. 2005). Additionally, some species also develop on mature basidiomes of Irpexlacteus (Boidin and Gilles 1982), as well as on soil and underground timber (Anon 1969).

Up to now, the genus of about 84 species have been accepted globally in Index Fungorum and MycoBank (Lentz 1967; Jung 1995; Langer et al. 2000a; Hagara 2001; Bernicchia et al. 2010; Saitta et al. 2011; Bates et al. 2017; Kalinina et al. 2020; Ram et al. 2021). There have been recorded about 35 species in Europe, 28 in North America, 26 in Latin America, 25 in Africa, 20 in Oceania, and 23 in Asia (Dritter 1809; Anon 1969; Holubová-Jechová 1969; Pouzar and Holubová-Jechová 1969; Holubová-Jechová 1980; Boidin and Gilles 1982; Boidin and Gilles 1988; Langer 1994; Greslebin and Rajchenberg 2003; Parmasto et al. 2004; Hjortstam et al. 2005; Bates et al. 2017; Buyck et al. 2017; Hyde et al. 2019; Vondrák et al. 2023). So far, 15 species of Botryobasidium have been reported from China, and most of them were distributed in the north temperate to subtropical zones (Dai 2011; Liu et al. 2024; Zhou et al. 2024).

During the surveys of lignicolous fungi in Yunnan Province, Southwestern China, several Botryobasidium specimens were collected from the mixed forests. The subsequent research by morphology and molecular phylogeny indicates that these specimens represent several undescribed species. The phylogenetic positions and the relationships of these species among Botryobasidium were clarified based on ITS + LSU dataset, and descriptions of these species with line drawings were provided in this study.

Materials and methods

Morphological study

The voucher specimens are deposited in the herbarium of the Institute of Applied Ecology, Chinese Academy of Sciences (IFP). Macromorphological characteristics were examined using a Nikon SMZ 645 (Tokyo, Japan) stereo microscope and the color descriptions refer to Kornerup and Wanscher (1981). Microscopical structures were checked using hand-cut sections stained with Cotton blue, Melzer’s reagent, and 3% KOH, and line drawings were prepared using a Nikon Eclipse 80i microscope (Nikon Corporation, Japan) with the aid of a drawing tube. The surface morphology for the basidiospores was observed with a field emission scanning electron microscope (SEM5000S, CIQTEK Co., Ltd.) at an accelerating voltage of 3 kV. The working distance was 9.62 mm. A thin layer of gold was plated on the sample to enhance the conductivity. Basidiospores were measured based on the front and back side view; the apex was excluded from the spore measurements. The following abbreviations are used: L = mean spore length, W = mean spore width, Q = L/W ratio, n (a/b) = number of spores (a) measured from number of specimens (b). Cotton blue (CB) was employed as a fitting medium to identify cyanophilous. Potassium hydroxide solution (KOH) was used to detect changes in hyphae, gloeocystidia, and encrusted. Melzer’s reagent (IKI) was used to determine amyloidity and dextrinoidity.

DNA extraction, PCR amplification, and DNA sequencing

According to the manufacturer’s instructions, the Fungal Fast Non-Toxic DNA Extraction Kit (Demeter Biotech Co., Ltd, Beijing, China) was employed to extract the sample’s total DNA and amplified by the polymerase chain reaction (PCR). The internal transcribed spacer (ITS) regions were amplified with the primers ITS1 and ITS4 (White et al. 1990), and the procedure was an initial denaturation at 95 °C for 3 min, followed by 34 cycles at 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, with a final extension at 72 °C for 5 min. The large subunit of nuclear ribosomal RNA gene (LSU) was amplified with the primers LR0R and LR7 (Vilgalys and Hester 1990), and the procedure involved an initial denaturation at 95 °C for 3 min, followed by 34 cycles at 95 °C for 30 s, 50 °C for 30 s, and 72 °C for 1 min, the procedure ended with an extension at 72 °C for 5 min.

DNA sequencing was conducted at the Beijing Genomics Institute (BGI), and the sequences were assembled using Geneious v.9.0.2 (Kearse et al. 2012). The generated sequences were verified and controlled to ensure their quality and integrity, and uploaded to GenBank (Table 1).

Table 1.

Species and GenBank numbers used in phylogenetic analysis in this study.

Species name ITS LSU Specimen No. Substrate Country References
Botryobasidiumacanthosporum L.J. Zhou & H.S. Yuan PP229497 / Yuan16326 on fallen angiosperm branch China Present study
B.acanthosporum PP229511 / Yuan17989 on bark of angiosperm China Present study
B.acanthosporum PP229512 PP218361 Yuan18083 on fallen trunk of Abies China Present study
B.acanthosporum PP229517 / Yuan18128 on fallen trunk of Abies China Present study
Botryobasidiumaureum Parmasto AJ389783 / GEL 2910 / Germany Langer et al. 2000b
B.botryosum (Bres.) J. Erikss. DQ267124 DQ089013 AFTOL-ID 604 / USA AFTOL Database
B.candicans J. Erikss. KP814200 / UC2022893 on litter or well decayed wood in pinaceous forest USA Rosenthal et al. 2017
B.cf.subcoronatum KP814216 / UC2022856 on litter or well decayed wood in pinaceous forest USA Rosenthal et al. 2017
B.cf.subcoronatum KP814322 / UC2022917 on litter or well decayed wood in pinaceous forest USA Rosenthal et al. 2017
B.coniferarum S.L. Liu & L.W. Zhou PP209210 PP218367 Yuan18440 on fallen gymnosperm trunk China Present study
B.coniferarum OR557262 OR527286 LWZ20171016-15 on fallen branch of Pinus China Liu et al. 2024
B.coniferarum OR557259 OR527282 LWZ20210928-3 on fallen branch of Pinus China Liu et al. 2024
B.conspersum J. Erikss. DQ911612 DQ521414 PBM 2747 (CUW) / USA AFTOL Database
B.conspersum OP163274 / FLAS-F-69114 / USA NCBI Database
B.conspersum / AY586657 GB/KHL11063 / Sweden Larsson et al. 2004
B.curtisii (Berk.) Hol.-Jech. EU118629 EU118629 KHL 12950GB / Costa Rica Larsson 2007
B.gossypirubiginosum Q. Zhou & C.L. Zhao OR668924 OR708665 CLZhao 26052 on fallen angiosperm branch China Zhou et al. 2024
B.incanum Q. Zhou & C.L. Zhao OR668923 OR708664 CLZhao 26697 on fallen angiosperm branch China Zhou et al. 2024
B.incanum PP209201 PP218357 Yuan17803 on fallen angiosperm branch China Present study
B.indicum (P.N. Singh & S.K. Singh) R. Kirschner & G. Langer PP209209 PP218363 Yuan18250 on root of Quercus China Present study
B.indicum ON406471 / CLZhao 21791 / China NCBI Database
B.indicum NR171230 NG070816 AMH:10054 dead bark of Leucaenaleucocephala India Hyde et al. 2019
B.indicum MK391496 MK391493 AMH:10054 dead bark of Leucaenaleucocephala India Hyde et al. 2019
B.intertextum (Schwein.) Jülich & Stalpers KP814540 / UC2022959 on litter or well decayed wood in pinaceous forest USA Rosenthal et al. 2017
B.intertextum AJ389782 / DAOM 197881 / Canada Langer et al. 2000b
B.isabellinum (Fr.) D.P. Rogers MZ159478 / K(M):181602 / UK NCBI Database
B.leptocystidiatum L.J. Zhou & H.S. Yuan PP209211 PP218178 Yuan17548 on fallen branch of Pinus China Present study
B.leptocystidiatum PP204173 PP218180 Yuan17557 on dead tree of Pinus China Present study
B.leptocystidiatum PP209200 PP218353 Yuan17706 on fallen angiosperm trunk China Present study
B.leptocystidiatum PP209197 PP218354 Yuan17708 on bark of living angiosperm tree China Present study
B.leptocystidiatum PP209198 PP218355 Yuan17709 on fallen angiosperm trunk China Present study
B.robustius Pouzar & Hol.-Jech. MH859491 MH871272 CBS:945.69 / Czech Vu et al. 2019
B.robustius PP436446 / HAY-F-004374 / USA NCBI Database
B.subcoronatum (Höhn. & Litsch.) Donk EU118607 EU118607 KHL s.n. (GB) / Sweden Larsson 2007
B.subcoronatum MH211720 FLAS-F-61064 / USA NCBI Database
B.subcoronatum DQ200924 AY647212 AFTOL-ID 614 / USA Matheny et al. 2007
B.subovalibasidium. L.J. Zhou & H.S. Yuan PP209199 PP218152 Yuan16439 on fallen trunk of Hippophaerhamnoides China Present study
B.subovalibasidium PP209196 PP218362 Yuan18179 on fallen trunk of Abies China Present study
B.tubulicystidium G. Langer OL436769 / DK14_139 / USA NCBI Database
B.vagum (Berk. & M.A. Curtis) D.P. Rogers OR680661 / personal:Alden Dirks:ACD0672 / USA Zhou et al. 2024
B.vagum OR471092 / TENN:075258 on Pinus USA Zhou et al. 2024
B.yunnanense Q. Zhou & C.L. Zhao OR668925 OR708666 CLZhao 24877 on fallen angiosperm branch China Zhou et al. 2024
Suillosporiumcystidiatum (D.P. Rogers) Pouzar MN937573 MN937573 VS3830 On Piceajezoensisvar.jezoensis Russia NCBI Database
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Phylogenetic analyses

Suillosporiumcystidiatum (D.P. Rogers) Pouzar (Botryobasidiaceae) was chosen as the outgroup according to the result of sequence BLAST in NCBI database, ensuring that it has suitable phylogenetic distances from other species in Botryobasidium. The concatenated datasets of ITS and LSU sequences of the species in Botryobasidiaceae were used to infer the molecular phylogeny. The ITS and LSU sequences were aligned separately using MEGA v.7.0 (Kumar et al. 2016). Maximum likelihood (ML) analysis was done using RAxML v.1.5b2 (Silvestro and Michalak 2012) with non-parametric bootstrapping of 500 replicates under the GTRGAMMA model. A Bayesian inference (BI) was also performed for the same data sets using MrBayes 3.2.6 (Ronquist et al. 2012). A substitution model was selected in PhyloSuite v1.2.2 (Zhang et al. 2020). The Bayesian information criterion (BIC) values under each model were compared and the model with the lowest BIC value was selected. Two parallel analyses were then run in MrBayes for 2 million generations, with 4 chains each, sampling every 500 generations. Burn-in trees (initial 25%) were discarded for each run and posterior probabilities of the matrix were determined by calculating a majority-rule consensus tree generated from the post-burnin trees by the MCMC runs using the sump of MrBayes. The phylogenetic trees were visualized using FigTree v1.4.3 (Rambaut 2016). Branches that received bootstrap support for ML (ML-BS) ≥ 70% and BI (BPP) ≥ 0.95 were considered significantly supported, respectively. The datasets were deposited in TreeBASE (www.treebase.org/treebase-web/, study no. 31569).

Results

Phylogeny

The ITS dataset consists of 39 sequences representing 20 taxa of Botryobasidium, and a sample of Suillosporiumcystidiatum as the outgroup. The ITS sequence had an aligned length of 661 base pairs (bp), of which 321 were parsimony-informative, 75 were singleton sites, 265 were constant sites. The Bayesian analysis had an average standard deviation of split frequencies = 0.004148, and a 50% majority-rule consensus phylogram was generated. The best model was GTR + F + G4 [lset nst = 6, rates = Gamma, Ngammacat = 4, prset statefreqpr = dirichlet (1, 1, 1, 1)]. The ITS + LSU dataset consists of 40 sequences representing 20 taxa of Botryobasidium, and a sample of Suillosporiumcystidiatum as the outgroup. The ITS + LSU dataset had an aligned length of 1502 bp (including 663 bp of ITS and 839 bp of LSU), of which 434 were parsimony-informative, 158 were singleton sites, 910 were constant sites. The Bayesian analysis had an average standard deviation of split frequencies = 0.005929, and a 50% majority-rule consensus phylogram was generated. The best model was GTR + F + I + G4 [lset nst = 6, rates = invgamma, Ngammacat = 4, prset statefreqpr = dirichlet (1, 1, 1, 1)].

In the phylogenetic tree based on ITS dataset (Fig. 1), four specimens of B.acanthosporum formed a clade (ML 100%/BPP 1), and grouped with B.incanum, B.vagum, and B.isabellinum with full support (ML 100%/BPP 1). Two specimens of B.leptocystidiatum formed a clade with full support (ML 100%/BPP 1). The remaining two specimens of B.subovalibasidium formed a clade, and clustered with B.aureum, B.botryosum and B.candicans with strong support (ML 95%/BPP 1).

Figure 1.

Figure 1.

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Phylogram of Botryobasidium resulting from a maximum likelihood analysis based on ITS sequence. Maximum likelihood bootstrap values (ML, ≥ 70%; left) and Bayesian posterior probabilities (BPP, ≥ 0.95; right) are given at the nodes. New species are in yellow background.

In the phylogenetic trees based on ITS + LSU dataset (Fig. 2), the branches to which two new species belong swapped positions, but the taxonomic positions of these three new species and the relationships with their sibling species are no discrepancy. Moreover, the support of the branches to which two new species belong, B.acanthosporum and B.subovalibasidium, was strengthened (ML 98%/BPP 1). Thus, the phylogenetic analyses revealed the taxonomic positions of these three new species.

Figure 2.

Figure 2.

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Phylogram of Botryobasidium resulting from a maximum likelihood analysis based on ITS + LSU. Maximum likelihood bootstrap values (ML, ≥ 70%; left) and Bayesian posterior probabilities (BPP, ≥ 0.95; right) are given at the nodes. New species are in yellow background.

Taxonomy

. Botryobasidium acanthosporum

L.J. Zhou & H.S. Yuan sp. nov.

6DFEAACF-5B38-5AFC-817D-2A02BBC84131

Fungal Names: FN 572031

Figs 3A, B , 4 , 5

Figure 3.

Figure 3.

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The habitats and basidiomes of three new species of BotryobasidiumA, BB.acanthosporum (holotype Yuan 18083) C, DB.leptocystidiatum (holotype Yuan 17708) E, FB.subovalibasidium (holotype Yuan 18179).

Figure 4.

Figure 4.

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Microscopic features of Botryobasidiumacanthosporum (drawn from holotype Yuan 18083) A a section through basidiome B basidiospores C basidia D basidioles E leptocystidia. Scale bars: 10 μm.

Figure 5.

Figure 5.

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SEM of basidiospores of Botryobasidiumacanthosporum species (holotype Yuan 18083).

Diagnosis.

Differed from other Botryobasidium species in having arachnoid basidiome with attached granules, clavate to subcylindrical cystidia, and subglobose to globose basidiospores with blunt spines up to 4 µm long.

Type.

China • Yunnan Province, Diqing Prefecture, Pudacuo National Park, 27°53'54"N, 99°57'04"E, on fallen trunk of Abies, 14 August 2023, Yuan 18083 (IFP 19972).

Etymology.

acanthosporum (Lat.), referring to the spore with spines.

Description.

Basidiomes: annual, adnate and resupinate, fluffy, pellicular, arachnoid with attached granules, 50–150 μm thick, adherent to the substrate and separates easily when wet. Hymenophoral surface smooth, greyish white to yellowish white (1B2–4B2) when fresh, pale yellow to dark yellow (3A3–4C8) when dry. Sterile margin often indeterminate and not differentiated.

Hyphal structure: hyphal system monomitic; generative hyphae simple septate, thin- to slightly thick-walled; tissues unchanged in KOH.

Subiculum: subicular hyphae colorless, thick-walled, frequently branched at right angles, cyanophilous, inamyloid, loosely interwoven, 7–12 μm in diam. Subhymenial hyphae colorless, thin-walled, acyanophilous, inamyloid, 7–11 μm in diam.

Cystidia: clavate to tubular, infrequent, smooth, thin-walled, colorless, simple septate, apically obtuse, acyanophilous, inamyloid, unchanged in KOH and distilled water, 26–37(–64) × 7–10 μm.

Basidia: clavate to subcylindrical, smooth, thin-walled, with 2 sterigmata, simple septate, acyanophilous, inamyloid, unchanged in KOH and distilled water, 14.5–20 × 8–10 μm.

Basidiospores: subglobose to globose, aculeate, slightly thick- to thick-walled, colorless, cyanophilous, inamyloid, unchanged in KOH and distilled water, 8–10(–10.3) × 8–10 μm (exclude spines), L = 9.25 μm, W = 8.92 μm, Q = 1.0–1.13 (n = 60/2); spines with apically obtuse, usually isolated, sometimes grouped in 2, up to 4 µm long.

Chlamydospores absent and anamorph not seen.

Ecology and distribution.

Growing in mixed forests dominated by Abies and a small number of Picea, Quercus, and other angiosperm trees. So far, known from Yunnan Province and Xizang Autonomous Region, China.

Additional specimens examined.

China • Xizang Autonomous Region, Bomi County, Yigong Tea Farm, 30°07'55"N, 95°01'05"E, on fallen angiosperm branch, 24 October 2021, Yuan 16326 (IFP 19970; paratype) • Yunnan Province, Diqing Prefecture, Baimaxueshan National Nature Reserve, 28°18'19"N, 99°08'57"E, on bark of angiosperm, 13 August 2023, Yuan 17989 (IFP 19971) • Pudacuo National Park, 27°53'56"N, 99°57'16"E, on fallen trunk of Abies, 14 August 2023, Yuan 18128 (holotype IFP 19973).

. Botryobasidium leptocystidiatum

L.J. Zhou & H.S. Yuan sp. nov.

CB91A294-5EE4-554F-A6B8-0A8E56DB662C

Fungal Names: FN 571970

Figs 3C, D , 6

Figure 6.

Figure 6.

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Microscopic features of Botryobasidiumleptocystidiatum (drawn from holotype Yuan 17708) A a section through basidiome B basidiospores C basidioles D basidia E leptocystidia. Scale bars: 10 μm.

Diagnosis.

Differed from other Botryobasidium species in having tubular cystidia and clamped in all hyphae.

Type.

China • Yunnan Province, Lincang City, Wulaoshan National Forest Park, 23°54'47"N, 100°10'53"E, on bark of living angiosperm tree, 9 August 2023, Yuan 17708 (holotype IFP 019955).

Etymology.

leptocystidiatum (Lat.), referring to the leptocystidia.

Description.

Basidiomes: annual, resupinate, effuse, pellicular, fluffy to arachnoid, 100–150 μm thick, adherent to the substrate and not easily separated. Hymenophoral surface smooth, greyish white (1B1–30B1) to smoky grey (3C2) when fresh, greyish white (1B1–30B1) to ivory (4B3) when dry; margin often indeterminate and not differentiated.

Hyphal structure: hyphal system monomitic; generative hyphae clamped, thin- to slightly thick-walled; tissues unchanged in KOH.

Subiculum: subicular hyphae colorless, slightly thick-walled, sparsely branched at right angles, cyanophilous, inamyloid, loosely interwoven, 7–10 μm in diam. Subhymenial hyphae colorless, thin-walled, frequently branched at right angles, acyanophilous, inamyloid, loosely interwoven, 4–7 μm in diam.

Cystidia: tubular, infrequent, smooth, thin-walled, colorless, apically obtuse, basal clamped, without additional septate, acyanophilous, inamyloid, unchanged in KOH and distilled water, 21.5–77 × 4–7.5 μm.

Basidia: ordered by botryose cluster, subcylindrical, smooth, thin-walled, usually with 6 sterigmata, occasionally with 7 sterigmata, basal clamped, acyanophilous, inamyloid, unchanged in KOH and distilled water, 10.5–15 × 7–8 μm.

Basidiospores: subnavicular to navicular, smooth, thin-walled, colorless, occasionally a few stuck together, acyanophilous, inamyloid, unchanged in KOH and distilled water, (6–)6.5–7.8(–8.1) × (2.8–)2.9–3.7(–3.9) μm, L = 7.2 μm, W = 3.1 μm, Q = 1.92–2.5 (n = 120/3).

Chlamydospores absent and anamorph not seen.

Ecology and distribution.

Growing in mixed forests dominated by Pinus and a small number of Fagaceae trees. So far only known from Yunnan Province, China.

Additional specimens examined.

China • Yunnan Province, Lincang City, Wulaoshan National Forest Park, 23°54'47"N, 100°10'53"E, on fallen branch of Pinus, 8 August 2023, Yuan 17548 (IFP 019952; paratype) • on dead tree of Pinus, 8 August 2023, Yuan 17557 (IFP 019953) • on fallen angiosperm trunk, 9 August 2023, Yuan 17706 (IFP 019954), Yuan 17709 (IFP 019956).

. Botryobasidium subovalibasidium

L.J. Zhou & H.S. Yuan sp. nov.

4FB91763-7382-51E3-843A-92D77BEA14CE

Fungal Names: FN 571974

Figs 3E, F , 7

Figure 7.

Figure 7.

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Microscopic features of Botryobasidiumsubovalibasidium (drawn from holotype Yuan 18179) A a section through basidiome B basidiospores C basidioles D basidia E secondary spores F chlamydospores G conidiophores. Scale bars: 10 μm.

Diagnosis.

Differed from other Botryobasidium species in having effuse, yellowish to ivory basidiomes, subovoid to ovoid basidia, ellipsoid chlamydospores.

Type.

China • Yunnan Province, Diqing Prefecture, Pudacuo National Park, 27°83'67"N, 99°95'76"E, Alt. 3655 m, on fallen trunk of Abies, 15 August 2023, Yuan 18179 (holotype IFP 019957).

Etymology.

subovalibasidium (Lat.), referring to the subovoid basidia.

Description.

Basidiomes: annual, resupinate, effuse, fluffy, 100–200 µm thick, adherent to the substrate and not easily separated. Hymenophoral surface smooth, greyish white (1B1–30B1) to ivory (4B3) when fresh, pale yellow (4A3) to greyish yellow (4B5) when dry; margin not differentiated, distinct.

Hyphal structure: hyphal system monomitic; generative hyphae simple septate, thin- to slightly thick-walled; tissues unchanged in KOH.

Subiculum: subicular hyphae colorless, slightly thick-walled, frequently branched, cyanophilous, inamyloid, loosely interwoven, (7–)8–11.5 μm in diam. Subhymenial hyphae colorless, thin-walled, moderately branched, acyanophilous, inamyloid, loosely interwoven, 5–8.5 μm in diam.

Cystidia: absent.

Basidia: subovoid to ovoid, smooth, thin-walled, with 4–6 sterigmata, basal simple septate, acyanophilous, inamyloid, unchanged in KOH and distilled water, (12–)14–18 × 9–10 µm.

Basidiospores: navicular to suburniform, smooth, thin-walled, colorless, occasionally stuck together, acyanophilous, inamyloid, unchanged in KOH and distilled water, (5.7–)7–9.8(–10) × (3.2–)3.7–5(–5.1) µm, L = 8.3 µm, W = 4.2 µm, Q = 1.53–2.5 (n = 60/1).

Chlamydospores: orange-yellow, ellipsoid, smooth, thick-walled, cyanophilous, inamyloid, unchanged in KOH, unchanged in distilled water, 17–21(–22) × (9–)10–11 µm, L = 18.5 µm, W = 10.3 µm, Q = 1.50–2.1 (n = 60/2).

Ecology and distribution.

Growing in mixed forests dominated by Abies and a small number of Picea, Quercus, and other angiosperm trees. So far, known from Yunnan Province and Xizang Autonomous Region, China.

Additional specimen examined.

China • Xizang Autonomous Region, Bomi County, on fallen trunk of Hippophaerhamnoides, 26 October 2021, Yuan 16439 (IFP 019951; paratype).

Discussion

In this study, three new species of Botryobasidium collected from Southwestern China are described based on morphological characteristics and phylogenetic analyses combining ITS and LSU sequences. The molecular phylogenetic analyses showed moderate to high support in the deeper nodes and at the species level which is consistent with the previous study (Cao et al. 2021; Zhou et al. 2024).

The phylogenetic trees show that B.acanthosporum is closely linked to B.incanum, B.isabellinum and B.vagum (Figs 1, 2). B.acanthosporum resembles B.incanum and B.vagum in having pellicular and greyish basidiomes. However, the new species is unique by having spine-ornamented basidiospores. B.acanthosporum is similar to B.isabellinum in having yellowish basidiomes and spine-ornamented basidiospores, but B.isabellinum differs from the new species by having narrower subhymenial hyphae (6–8 μm), absence of cystidia, longer basidia (15–25 × 8–10 μm) and smaller globose basidiospores (7–10 μm) (Bernicchia and Gorjón 2010). In morphology, B.bondarcevii resembles B.acanthosporum in having pellicular and slightly yellow to dark yellow basidiomes, and spine-ornamented basidiospores, but B.bondarcevii can be distinguished by having bigger basidia (18–23 × 9–11 μm vs. 14.5–20 × 8–10 μm), ellipsoid basidiospores (Xiong et al. 2007).

In the phylogenetic trees (Figs 1, 2), B.leptocystidiatum grouped together with B.subcoronatum. Morphologically, they share similar characteristics in having thin and whitish to pale yellow basidiomes, clamped generative hyphae with frequently vertical branches, and subnavicular basidiospores. Nevertheless, B.subcoronatum differs from B.leptocystidiatum by the absence of cystidia, longer basidia and slightly narrower basidiospores (6–8 × 2.5–3 µm vs. 6.5–7.8 × 2.9–3.7 µm). Moreover, B.leptocystidiatum and B.sassofratinoense are similar in having greyish-white to yellow basidiomes and clamped hyphae. But B.sassofratinoense can be differentiated by having wider subhymenial hyphae (5–8 μm vs. 4–7 μm) and subicular hyphae (10–12 μm vs. 7–10 μm), shorter cystidia (28–45 × 5–7 μm vs. 21.5–77 × 4–7.5 μm), longer basidia (18–25 × 6–8.5 μm vs. 10.5–15 × 7–8 μm) and slightly bigger basidiospores (6.5–7.8 × 2.9–3.7 μm vs. 7–8.5 × 3.5–4.5 μm) (Bernicchia and Gorjón 2010).

Botryobasidiumsubovalibasidium has an adjacent phylogenetic relationship with B.aureum, B.botryosum and B.candicans in the phylogenetic trees (Figs 1, 2). In morphology, they exhibit some similarities in having whitish to yellowish basidiomes and absence of cystidia (Breitenbach and Kränzlin 1986; Bernicchia and Gorjón 2010). However, B.aureum is distinguished from the new species by having narrower subhymenial hyphae (4–6 µm) and subicular hyphae (5–10 µm) and smaller basidia (12–18 × 6–8 µm). Botryobasidiumcandicans differs from the new species by having narrower subhymenium and subicular hyphae, slightly smaller basidiospores (6–8 × 3–4 µm vs. 7–9.8 × 3.7–5.0 µm), and smaller chlamydospores (15–17 × 9–12 µm vs. 17–21 × 10–11 µm) (Bernicchia and Gorjón 2010). Botryobasidiumbotryosum can be separated from the new species by larger basidia, bigger basidiospores (8.5–11 × 4.4–5.5 μm vs. 7–9.8 × 3.7–5.0 µm), and absence of conidiospores (Jülich 1978). Moreover, B.subovalibasidium and B.danicum are similar in having greyish to yellowish basidiomes, absence of cystidia, and basidia with 4–6 sterigmata. However, B.danicum is distinct from the new species by longer basidiospores (12–14 × 3.0–5.0 µm vs. 7–9.8 × 3.7–5.0 µm), bigger basidia (15–20 × 8–12 µm vs. 14–18 × 9–10 µm) and absence of anamorphic spores (Bernicchia and Gorjón 2010).

Key to known 18 species of Botryobasidium in China

1 Basidiospores with spines 2
– Basidiospores smooth 4
2 Basidiospores ellipsoid, 7–9 × 5–6.3 µm B.bondarcevii
– Basidiospores globose 3
3 Basidiospores 7–10 µm, spines up to 1–3 µm, basidia with 4 sterigmata B.isabellinum
– Basidiospores 8–10 µm, spines up to 4 µm, basidia with 2 sterigmata B.acanthosporum
4 Conidia absent 5
– Conidia present 15
5 Hyphae with clamps at least in a part of basidiome 6
– Hyphae without clamps 9
6 Clamps present on all septa 7
– Both clamps and simple septa present 8
7 Basidiospores navicular, 6–7 × 2.5–3 µm; cystidia absent B.subcoronatum
– Basidiospores subnavicular to navicular, 6.5–7.8 × 2.9–3.7 µm; cystidia present B.leptocystidiatum
8 Clamps often present in subiculum and subhymenium B.angustisporum
– Clamps often absent in subiculum B.intertextum
9 Basidiospores navicular 10
– Basidiospores not navicular 12
10 Basidiospores 7–8 × 3–3.5 µm B.coniferarum
– Basidiospores more than 8 µm long 11
11 Basidiospores 9–10 × 3.5–5 µm; basidia cylindrical, 9–16 × 7–9 µm B.subbotryosum
– Basidiospores 8–12 × 4.5–6 µm; basidia clavate to subcylindrical, 20–25 × 8–12 µm B.vagum
12 Basidiospores obliquely ovoid, apically obtuse 13
– Basidiospores not ovoid 14
13 Basidiospores 7.5–12 × 3.5–5 µm B.obtusisporum
– Basidiospores 5–8 × 2.5–3.5 µm B.pruinatum
14 Basidiospores subglobose, 14–17.5 × 13–15.5 µm B.gossypirubiginosum
– Basidiospores ellipsoid, 6.5–8.5 × 3.5–5 µm B.incanum
15 Conidia ellipsoid 16
– Conidia not ellipsoid 17
16 Conidia 13–22 × 9–12 µm; basidia ellipsoid to obovate, 12–15 × 6–8 µm B.conspersum
– Conidia 17–21 × 10–11 µm; basidia subovoid to ovoid, 14–18 × 9–10 µm B.subovalibasidium
17 Conidia subglobose to citriform, 15–20 × 8–10 µm B.candicans
– Conidia subglobose to globose, 11.5–14.5 × 9.5–10.5 µm B.yunnanense
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Supplementary Material

XML Treatment for Botryobasidium acanthosporum
XML Treatment for Botryobasidium leptocystidiatum
XML Treatment for Botryobasidium subovalibasidium

Citation

Zhou L-J, Li X-L, Yuan H-S (2024) Three new wood-inhabiting fungi of Botryobasidium (Cantharellales, Basidiomycota) from subtropical forests of Southwestern China. MycoKeys 109: 337–354. https://doi.org/10.3897/mycokeys.109.133325

Footnotes

Lin-Jiang Zhou and Xue-Long Li contributed equally to this work.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This research was supported by the National Natural Science Foundation of China (Project Nos. U2102220 & 31970017) and the CAS Key Laboratory Annual Project.

Author contributions

Investigation and writing draft: LJZ. Data measurement and analysis: XLL. Conceptualization and supervision: HSY. All authors contributed to the article and approved the submitted version.

Author ORCIDs

Lin-Jiang Zhou https://orcid.org/0000-0003-2665-6959

Xue-Long Li https://orcid.org/0009-0006-3948-0234

Hai-Sheng Yuan https://orcid.org/0000-0001-7056-140X

Data availability

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

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Associated Data

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

Supplementary Materials

XML Treatment for Botryobasidium acanthosporum
mycokeys.109.133325-treatment1.xml (13.2KB, xml)
XML Treatment for Botryobasidium leptocystidiatum
mycokeys.109.133325-treatment2.xml (11.5KB, xml)
XML Treatment for Botryobasidium subovalibasidium
mycokeys.109.133325-treatment3.xml (10.8KB, xml)

Data Availability Statement

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

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