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Research Article
Three new species of Dicephalospora (Leotiomycetes, Helotiales) from Northern Thailand and Southwestern China
expand article infoLe Luo‡§|, Kandawatte Wedaralalage Thilini Chethana§, Qi Zhao‡, Vinodhini Thiyagaraja‡, Kitiphong Khongphinitbunjong§, Fatimah Al-Otibi, Kevin D. Hyde§‡
‡ Chinese Academy of Sciences, Kunming, China
§ Mae Fah Luang University, Chiang Rai, Thailand
| Guiyang Institute of Humanities and Technology, Guiyang, China
¶ King Saud University, Riyadh, Saudi Arabia

Corresponding author: Kandawatte Wedaralalage Thilini Chethana ( kandawatte.thi@mfu.ac.th )

Corresponding author: Kevin D. Hyde ( kdhyde3@gmail.com )

Academic editor: Danushka Sandaruwan Tennakoon
© 2025 Le Luo, Kandawatte Wedaralalage Thilini Chethana, Qi Zhao, Vinodhini Thiyagaraja, Kitiphong Khongphinitbunjong, Fatimah Al-Otibi, Kevin D. Hyde.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Luo L, Chethana KWT, Zhao Q, Thiyagaraja V, Khongphinitbunjong K, Al-Otibi F, Hyde KD (2025) Three new species of Dicephalospora (Leotiomycetes, Helotiales) from Northern Thailand and Southwestern China. MycoKeys 115: 67-85. https://doi.org/10.3897/mycokeys.115.143994
Open Access

Abstract

Dicephalospora is a discomycetous genus belonging to the family Helotiaceae (Helotiales). The genus currently comprises 19 species. Among them, 17 species have been reported from Asia, of which 14 were reported from China. During a survey of Leotiomycetes, we collected six specimens of Dicephalospora species from southwest China and northern Thailand. The samples were examined based on the phylogenetic analyses of LSU and ITS sequence data, along with morphological characterization, and our results demarcate three new Dicephalospora species, viz., D. tengyueica, D. maetaengica and D. menghaica. Dicephalospora tengyueica sp. nov. formed a distinct clade closely related to D. rufocornea, D. irregularis and D. sagerae, whereas D. maetaengica sp. nov. clustered closer to D. menghaica sp. nov. with 100% ML and 1.00 BIPP statistical support. All three species have similar apothecia sizes but differ in their coloration. Dicephalospora tengyueica has aseptate ascospores, while D. menghaica and D. maetaengica have 7-septate ascospores. The paraphyses in D. menghaica are branched, while they are unbranched in the other two species. An updated key to the known species of Dicephalospora is also provided.

Key words:

3 new species, Helotiaceae, morphology, phylogeny, taxonomy

Introduction

Dicephalospora belongs to Helotiaceae (Helotiales) and was described by Spooner in 1987, with D. calochroa (Syd. and P. Syd.) Spooner as the type species (Spooner 1987; Hyde et al. 2024). The taxonomy of the Helotiales has undergone multiple revisions, driven largely by advances in molecular phylogenetics (Ekanayaka et al. 2019; Johnston et al. 2019). Two gene regions, ITS and LSU, have been primarily used in the phylogenetic analyses of Helotiales, further supporting the inclusion of Dicephalospora within Helotiaceae (Han et al. 2014; Zhao et al. 2016; Ekanayaka et al. 2019). Helotiaceae is the most heterogeneous family in Helotiales and is characterized by parallel hyphae, angular to isodiametric cells, interwoven hyphae, and margins and flanks that may be covered with hairs or completely absent (Groves 1968; Korf 1973; Wijayawardene et al. 2022). The taxonomy of Helotiales, including Helotiaceae, has been widely researched and debated. Notably, a multi-gene phylogenetic analysis incorporating up to 15 genes across 279 specimens has provided a more resolved, refined classification framework for Leotiomycetes, including members of Helotiaceae (Johnston et al. 2019). After multiple revisions, a total of 26 genera are now accepted within the family, many of which are saprobic on plant material (Ekanayaka et al. 2019; Wijayawardene et al. 2022).

Morphologically, Dicephalospora species are distinct due to their erumpent or superficial, stipitate apothecia, which can vary in color from yellow and orange to red and blackish. The ectal excipulum comprises "textura prismatica" cells with refractive walls, while the medullary excipulum consists of cells of "textura intricata". The asci are filiform and show variable reactions in Melzer’s reagent (J+ or J-). The paraphyses are straight or slightly curved at the apex, and the ascospores are hyaline, sub-ellipsoid to fusoid, guttulate, and often have a mucilaginous cap at the poles (Hosoya et al. 1999; Zhuang et al. 2016). The taxa are predominantly distributed in Asia (Phutthacharoen et al. 2022), with 17 of 19 listed species reported from this region (Luo et al. 2024). Exceptionally, D. chrysotricha is reported from New Zealand (Zheng and Zhuang 2019), D. sagerae is reported from Australia, and D. calochroa shows extended distribution and is reported in China and Papua New Guinea (Spooner 1987). So far, 14 species (viz., D. albolutea, D. aurantiaca, D. calochroa, D. contracta, D. damingshanica, D. dentata, D. huangshanica, D. phaeoparaphysis, D. pinglongshanica, D. rufocornea, D. sessilis, D. shennongjiana, D. xishuangbannaensis and D. yunnanica) have been reported from China (Phutthacharoen et al. 2022; Luo et al. 2024).

Hosoya et al. (1999) discovered dicephalosterol from the culture of D. rufocornea, which has a potential application for prostatic hypertrophy. However, the applications of Dicephalospora species are poorly explored due to the difficulty in obtaining pure cultures and their slow growth on artificial media, as well as their minimal biomass in nature (Zheng and Zhuang 2019).

Dicephalospora represents a morphologically distinct and phylogenetically significant genus within Helotiaceae. While challenges remain in cultivating these fungi and exploring their biochemical potential, ongoing molecular studies continue to advance our understanding of their taxonomy, ecology, and potential applications. During the investigation of Leotiomycetes, six collections of Dicephalospora were obtained from southwest China and northern Thailand. Morphological and phylogenetic analyses based on LSU and ITS data were performed to confirm their taxonomic placement, which revealed three new Dicephalospora species based on a polyphasic approach, along with an updated dichotomous key for the genus.

Material and methods

Collection and morphological examinations

Six specimens were collected from southwest China and northern Thailand, primarily in highly humid, natural broadleaf forests and protected areas with minimal human interference. The fruiting bodies were found on the surface of decaying wood and were photographed in the field before placing them in plastic containers for transport to the laboratory. The samples were dehydrated using a dehydrator set to a temperature between 25–30 °C. Further, the samples were examined using morphological and phylogenetic analyses. For detailed morphological examination, dried specimens were observed under a stereomicroscope (C-PSN, Nikon, Japan) with images captured using a digital camera (Canon EOS 70D, Japan) attached to the microscope. Free-hand sections of the dried specimens were mounted in water to observe microscopic characteristics such as apothecia, exciple, paraphyses, asci, and ascospores using a Nikon compound microscope (Nikon, Japan) with a DS-Ri2 camera. Sections were also treated with Melzer’s reagent for the iodine test (Tochihara and Hosoya 2022). Measurements of the microstructures were taken using the Tarosoft (R) Image Frame Work program v.0.97 (Tarosoft, Thailand). These measurements are presented in the format (a–) b–c(–d), where ‘a’ represents the minimum value, ‘d’ the maximum value, and ‘b–c’ the 90% confidence interval. The specimens were deposited in the Cryptogamic Herbarium of the Kunming Institute of Botany, Chinese Academy of Sciences (KUN-HKAS), and the Mae Fah Luang University Herbarium (Herb. MFLU). The Facesoffungi and the Index Fungorum numbers were obtained following the procedures outlined by Jayasiri et al. (2015) and Index Fungorum (2025), respectively. The morphological description and the phylogenetic tree of the new species were submitted to the Greater Mekong Subregion webpage (Chaiwan et al. 2021). Images used for figures were processed with Adobe Photoshop CS6 Extended version 13.0 × 64 (Adobe Systems, USA).

DNA extraction, PCR amplifications and sequencing

Genomic DNA was extracted from the dried apothecia using a TSP101 DNA extraction kit (TSINGKE, China). Following the latest studies (Phutthacharoen et al. 2022; Luo et al. 2024), LSU and ITS regions were subjected to PCR amplification, using the primers LR0R/LR5 (Vilgalys and Hester 1990) and ITS1/ITS4 (White et al. 1990; Gardes and Bruns 1993), respectively. The total volume of PCR amplifications was 25 μL, including 12.5 μL of 2X PCR G013 Taq MasterMix with Dye (Applied Biological Materials, Canada), 1 μL of each primer (10 μM), 2 μL of genomic DNA, and 8.5 μL of sterilized, distilled water. Amplifications were conducted under the following conditions: pre-denaturation at 95 °C for 5 minutes, followed by 35 cycles of denaturation at 95 °C for 20 seconds, annealing at 55 °C for 10 seconds, elongation at 72 °C for 20 seconds and final elongation at 72 °C for 7 minutes. Gel electrophoresis with 1% TAE and TSJ003 GoldView nucleic acid dye (TSINGKE, China) was used to test the obtained PCR products. Finally, the PCR products were sequenced by Tsingke Biotechnology Co., Ltd., Kunming, China. Newly produced sequences were deposited in the GenBank and the accession numbers were given in Table 1.

Table 1.
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The taxa included in the phylogenetic analysis along with their corresponding GenBank accession numbers. Newly identified taxa are in bold. Names with (T) indicate type specimens and ‘-’ denotes unavailable data in the GenBank.

Species Strain GenBank Accession No. Reference
ITS LSU
Amylocarpus encephaloides CBS 129.60 MH857920 MH869464 Vu et al. (2019)
Amylocarpus encephaloides 017cN KM272369 KM272361 Rämä et al. (2014)
Bryoscyphus dicrani M141 EU940183 EU940107 Stenroos et al. (2010)
Connersia rilstonii CBS 537.74 KJ755499 AF096189 Suh and Blackwell (1999)
Crocicreas amenti F-147481 FJ005093 FJ005124 Peláez et al. (2011)
Crocicreas cacaliae F-148706 FJ005107 FJ005126 Peláez et al. (2011)
Crocicreas cyathoideum MFLU 18-0698 MK584943 MK591970 Ekanayaka et al. (2019)
Crocicreas tomentosum MFLU 17-0082 MK584988 MK592008 Ekanayaka et al. (2019)
Cudoniella clavus AFTOL-ID 166 DQ491502 DQ470944 Spatafora et al. (2006)
Cyathicula microspora M267 EU940165 EU940088 Baral et al. (2009)
Dicephalospora albolutea HMAS 279693 MK425601 – Zheng and Zhuang (2019)
Dicephalospora aurantiaca MFLU 16-0591a MK584962 MK591988 Ekanayaka et al. (2019)
Dicephalospora chiangraiensis MFLU 21-0019 MZ241818 MZ241827 Phutthacharoen et al. (2022)
Dicephalospora chiangraiensis MFLU 21-0018(T) MZ241817 MZ241826 Phutthacharoen et al. (2022)
Dicephalospora chrysotricha PDD:91762 KF727411 – Unpublished
Dicephalospora chrysotricha PDD:58197 KF727409 – Unpublished
Dicephalospora chrysotricha PDD:93932 MH578487 – Unpublished
Dicephalospora chrysotricha PDD:81537 KF727410 OQ466391 Unpublished
Dicephalospora dentata 3093 KP204263 – Liu et al. (2016)
Dicephalospora huangshanica MFLU 18-1828 MK584979 MK591979 Ekanayaka et al. (2019)
Dicephalospora huangshanica KUS-F52405 JN033408 JN086711 Han et al. (2014)
Dicephalospora inthanonensis MFLU 22-0050(T) ON606312 ON604634 Phutthacharoen et al. (2022)
Dicephalospora inthanonensis MFLU 22-0053 ON606313 ON604635 Phutthacharoen et al. (2022)
Dicephalospora irregularis MFLU 22-0054(T) ON511117 ON514038 Phutthacharoen et al. (2022)
Dicephalospora maetaengica MFLU24-0330 (T) PQ481904 PQ481910 This study
Dicephalospora maetaengica MFLU24-0331 PQ481905 PQ481911 This study
Dicephalospora menghaica HMAS 135692 (T) PQ481908 PQ481914 This study
Dicephalospora menghaica HMAS 135690 PQ481909 PQ481915 This study
Dicephalospora rufocornea MFLU 16-1860 MK584989 MK592011 Ekanayaka et al. (2019)
Dicephalospora rufocornea MFLU 19-2083 MZ241816 MZ241825 Phutthacharoen et al. (2022)
Dicephalospora rufocornea TNS:F:36242 LC136911 – Unpublished
Dicephalospora rufocornea TNS:F:40155 LC136918 – Unpublished
Dicephalospora rufocornea MFLU 16-1858 MK584991 – Ekanayaka et al. (2019)
Dicephalospora rufocornea FCATAS5710 PP622049 – Unpublished
Dicephalospora sagerae BRIP 72428d NR_182617 – Tan and Shivas (2022)
Dicephalospora sessilis MFLU 18-1823(T) NR_163779 NG_068621 Ekanayaka et al. (2019)
Dicephalospora shennongjiana HMAS 279698 MK425606 – Zheng and Zhuang (2019)
Dicephalospora xishuangbannaensis HMAS 131157 OR948047 – Luo et al. (2024)
Dicephalospora xishuangbannaensis HMAS 131164(T) OR948048 – Luo et al. (2024)
Dicephalospora tengyueica HMAS 135691 (T) PQ481906 PQ481912 This study
Dicephalospora tengyueica HMAS 135694 PQ481907 PQ481913 This study
Dicephalospora yunnanica HMAS 279701 MK425609 – Zheng and Zhuang (2019)
Dicephalospora yunnanica HMAS 279700 MK425608 – Zheng and Zhuang (2019)
Dicephalospora yunnanica HMAS 61850 DQ986486 – Zheng and Zhuang (2019)
Endoscypha perforans PDD:102231 KF727424 MK039717 Unpublished
Glarea lozoyensis ATCC 20868(T) NR_137138 – Bills et al. (1999)
Glarea sp. C2B KX610435 – Yokoya et al. (2017)
Gloeotinia granigena CBS 417.50 – MH868212 Vu et al. (2019)
Hymenoscyphus fructigenus CBS 186.47 MH856211 MH867741 Vu et al. (2019)
Hymenoscyphus occultus KUS-F52847 KP068064 – Gross and Han (2015)
Hymenoscyphus pseudoalbidus Hokk_14 KJ511191 – Gross et al. (2014)
Hymenotorrendiella eucalypti PDD:70105 MH578483 – Unpublished
Hymenotorrendiella eucalypti CPC 11050(T) DQ195788 DQ195800 Zhao (2014)
Lanzia berggrenii ICMP:19614 KC164645 KC164640 Johnston and Park (2013)
Ombrophila violacea WZ0024 AY789366 AY789365 Wang et al. (2005)
Phaeohelotium epiphyllum TNS: F-40042 AB926061 AB926130 Zhao (2014)
Pirottaea palmicola PDD:60282 KM677208 – Unpublished
Pirottaea palmicola PDD:65971 KM677206 – Unpublished
Pleuroascus nicholsonii CBS 345.73 KJ755519 AF096196 Zhao (2014)
Roesleria subterranea CBS 339.96 EF060308 EF608074 Kirchmair et al. (2008)
Roesleria subterranea CBS 407.51 MH856922 – Vu et al. (2019)

Phylogenetic analyses

Newly generated DNA sequences were assembled using BioEdit v.7.2.5 (Hall 1999) to obtain consensus sequences. The concatenated sequences were used to search for the close relatives in the NCBI (Johnson et al. 2008). The closely related sequences were downloaded from GenBank following the recent papers (Phutthacharoen et al. 2022; Luo et al. 2024) (Table 1). Pleuroascus nicholsonii (CBS 345.73) and Connersia rilstonii (CBS 537.74) were selected as outgroup taxa. The phylogenetic analysis was conducted based on the datasets, including reference DNA sequences and newly generated DNA sequences using OFPT (Zeng et al. 2023) with the following protocol. Datasets of each gene region were first independently aligned with the ‘auto’ strategy (based on data size) by MAFFT (Katoh and Standley 2013) and trimmed with the ‘gappyout’ option (based on gaps’ distribution) by TrimAl (Capella-Gutiérrez et al. 2009). The best-fit nucleotide substitution models for each dataset were then selected based on the Bayesian information criterion (BIC) from twenty-two common DNA substitution models with rate heterogeneity by ModelFinder (Kalyaanamoorthy et al. 2017). All datasets were concatenated with partition information for the subsequent phylogenetic analyses. Maximum likelihood with 1000 replicates was performed using ultrafast bootstrap approximation (Hoang et al. 2018) with SH-like approximate likelihood ratio test (SH-aLRT) (Guindon et al. 2010) by IQ-TREE (Nguyen et al. 2015). The consensus tree was summarized based on the extended majority rule. Bayesian inference was performed with two parallel Metropolis-coupled (one ‘cold’ chain and three heated chains) Markov chain Monte Carlo runs by MrBayes (Ronquist et al. 2012), with trees sampling every 100th generations. The consensus tree was summarized after discarding the first 25% of samples when the average standard deviation of split frequencies fell below 0.01. The resulting trees were visualized in FigTree v1.4.3 (http://tree.bio.ed.ac.uk/software/figtree/). The polyphasic approach recommended by Chethana et al. (2021) was followed to establish the new species.

Results

Phylogenetic analysis

The data matrix consisted of LSU and ITS sequences, representing 62 taxa distributed across 17 genera of Helotiaceae with 1192 characters after trimming, including gaps (ITS: 404 bp and LSU: 788 bp). The matrix had 390 distinct alignment patterns, with 28.73% undetermined characters or gaps. Estimated parameters for the tree are as follows: total tree length (sum of branch lengths): 1.4788; sum of internal branch lengths: 0.9205 (62.2453% of tree length); gamma distribution shape parameter α = 0.7196. The best IQ-Tree with a final likelihood value of -7128.2117 is presented in Fig. 1. The tree topology inferred from ML analysis is similar to that of BI analysis. Index Fungorum (2025) listed 19 species epithets for Dicephalospora, and 15 of them are included in our phylogenetic analyses (Fig. 1), and four species (D. calochroa, D. damingshanica, D. pinglongshanica and D. phaeoparaphysis) are excluded due to a lack of molecular data. The phylogenetic result showed that Dicephalospora species clustered together and concurred with the previous studies (Zheng and Zhuang 2019; Phutthacharoen et al. 2022; Luo et al. 2024). Dicephalospora species formed a monophyletic clade separated from an assemblage of Endoscypha and Hymenotorrendiella taxa by 93% maximum likelihood bootstrap support (MLBS) and 1.00 Bayesian inference posterior probability (BIPP) support. The new species D. tengyueica formed a distinct clade closely related to D. rufocornea, D. irregularis, and D. sagerae, with 93% MLBS and 0.99 BIPP support (Fig. 1). The two new species, D. maetaengica and D. menghaica, were closely related, supported by 100% MLBS and 1.00 BIPP, and together formed a sister clade to D. albolutea, with 98% MLBS and 1.00 BIPP support (Fig. 1).

Figure 1. 

The IQ-Tree phylogram based on the combined LSU and ITS dataset. The MLBS ≥ 70% and BIPP ≥ 0.90 are shown at the nodes as MLBS/BIPP. The MLBS < 70% and BPP < 0.90 are expressed as a hyphen (“-”). Type strains are in bold. Names in red indicate isolates from the current study.

Taxonomy

Dicephalospora tengyueica L. Luo & K.D. Hyde, sp. nov.

Index Fungorum: IF902889
Facesoffungi Number: FoF16773
Fig. 2

Etymology.

The epithet “tengyueica” refers to the collection site, Tengyue street, where the holotype specimen was collected.

Holotype.

HKAS135691.

Description.

Saprobic on dead twigs. Sexual morph: Apothecia 1.2–3.2 mm diam., when dry arising solitary or gregarious in a small group, scattered on wood, centrally stipitate, superficial, orange when fresh, become light brown when dry. Stipe 0.9–2.3 mm height at the base, yellow when fresh, and light yellow to white when dry. Receptacle orange and discoid. Margins slightly rough, orange to dark orange. Disc slightly convex and orange. Ectal excipulum 30–66 µm (x̄ = 46 µm, n = 20), multi-layered, and thin-walled, with hyaline cells of textura globulosa. Medullary excipulum 21–48 µm (x̄ = 33 µm, n = 30), composed of thin-walled, hyaline, gelatinized cells of textura globulosa to porrecta, small cells condensed. Hymenium 84–171 µm (x̄ = 131 µm, n = 60), hyaline to yellowish, inner mixed with asci and paraphyses. Paraphyses 1.1–3.2 µm wide (x̄ = 1.9 µm, n = 30), at the terminal cell, filiform, numerous, lengths exceeding the asci, unbranched, aseptate, apical cells swollen and globose, filled with oil droplets. Asci (85–)90–118(–125) × 5.9–12.5 µm (x̄ = 102 × 9.5 µm, n = 50), eight-spored, unitunicate, cylindrical, and clavate, J- in Melzer’s reagent, tapered long stipitate base. Ascospores (17–)19–32(–35) × 2–3.5 µm (x̄ = 27.4 × 2.9 µm, n = 60), uniseriate to biseriate, fusiform, aseptate, both ends are filled with oil droplets of different sizes, some narrowed ends capped with a small and gelatinous collar. Asexual morph: Not observed.

Figure 2. 

Dicephalospora tengyueica (HKAS 135691, holotype) a fresh apothecia growing on dead wood b, c dried apothecia d, e vertical sections of an apothecia f excipulum g paraphyses h–k asci (j–k asci in Meltzer’s reagent) l ascospores. Scale bars: 5 mm (a); 2 mm (b, c); 100 µm (d, e); 50 µm (f); 20 µm (g–l).

Material examined.

China • Yunnan Province, Tengchong City, Tengyue Street, altitude 1749 m, on the decayed unidentified twigs, 19 August 2022, Le Luo, ly289 (HKAS 135691, holotype); • ibid., ly290 (HKAS 135694, paratype).

Notes.

Our specimens of D. tengyueica (HKAS 135691 and HKAS 135694) formed a distinct clade (93% MLBS and 0.99 BIPP), closely related to D. rufocornea, D. irregularis and D. sagerae (Fig. 1). However, the new species exhibit distinct morphological features in having orange to light brown apothecia, light yellow stipe, tapered long stipitate base, aseptate ascospores, and aseptate paraphyses. Dicephalospora rufocornea has red or reddish-orange apothecia, red stipe, and sessile base with septate paraphyses (Ekanayaka et al. 2019), whereas D. irregularis has sessile apothecia, asci arising from simple septa without basal protuberance, aseptate to septate ascospores that are wider than D. tengyueica (5.5–7.5 µm vs. 2–3.5 µm) (Phutthacharoen et al. 2022). The detailed morphological description is not available for D. sagerae (Tan & Shivas, 2022) but it differs from D. tengyueica in the ITS base pair comparison, which revealed 8.5% differences. Therefore, D. tengyueica is introduced here as a new species.

Dicephalospora maetaengica L. Luo & K.D. Hyde, sp. nov.

Index Fungorum: IF902890
Facesoffungi Number: FoF16774
Fig. 3

Etymology.

The epithet “maetaengica” refers to the collection site, MaeTaeng District, where the holotype specimen was collected.

Holotype.

MFLU 24-0330.

Description.

Saprobic on dead leaves. Sexual morph: Apothecia 1.5–3.3 mm diam., when dry arising solitary, uniseriate on the stem of the leaves, centrally long stipitate, superficial, yellow when fresh and dry. Stipe 1.2–2.5 mm height, slightly hyaline to light yellow at the base. Receptacle yellow and cupulate. Margins smooth, yellow to pale yellow. Disc slightly sunken and yellow. Ectal excipulum 23–44 µm (x̄ = 33 µm, n = 60), multi-layered, thin-walled, hyaline to light yellow or pale green cells of textura globulosa, slightly larger cells condensed. Medullary excipulum 28–61 µm (x̄ = 45 µm, n = 60), composed of thin-walled, hyaline to yellowish, gelatinized cells of textura porrecta to globulosa, small cells condensed. Hymenium 99–146 µm (x̄ = 123 µm, n = 60), hyaline to yellow, inner mixed with asci and paraphyses. Paraphyses 1.1–2.1 µm wide (x̄ = 1.6 µm, n = 60), filiform, numerous, lengths exceeding the asci, unbranched, aseptate, slightly swollen, small, globose at the apex. Asci (75–)81–111(–118) × 5.6–10.7 µm (x̄ = 99 × 8.5 µm, n = 50), eight-spored, unitunicate, cylindrical, clavate, amyloid (J+) having a rounded apex in Melzer’s reagent, tapered long stipitate base. Ascospores (12.5–)14.5–21.8(–22.5) × 2.4–4.2 µm (x̄ = 18.2 × 3.3 µm, n = 60), uniseriate to biseriate, fusiform, guttulate, 0–1–septate. Asexual morph: Not observed.

Figure 3. 

Dicephalospora maetaengica (MFLU 24-0330, holotype) a fresh apothecia growing on a dead wood b, c dried apothecia d, e vertical sections of the apothecia f excipulum g paraphyses and part of asci h–j asci (i–j asci in Meltzer’s reagent) k–l ascospores. Scale bars: 2 mm (a–c); 100 µm (d, e); 50 µm (f–j); 10 µm (k, l).

Material examined.

Thailand • Chiang Mai Province, MaeTaeng District, Pha Deng Village, Mushroom Research Center (MRC), altitude 863 m, on the decayed unidentified leaf, 18 August 2023, Le Luo, RB1 (MFLU 24-0330, holotype); • ibid., RB101 (MFLU 24-0331, paratype).

Notes.

Dicephalospora maetaengica formed a clade closer to D. menghaica with 100% MLBS and 1.00 BIPP support (Fig. 1). These two species grouped as a sister clade and together formed a closely related clade to D. albolutea with 98% MLBS and 1.00 BIPP (Fig. 1) bootstrap support. Dicephalospora maetaengica differs from D. menghaica by having larger (1.5–3.3 mm vs. 1–1.6 mm), yellow apothecia, long stipe (1.2–2.5 mm vs. 0.6–1 mm), slightly hyaline to light yellow at the base of stipe, 0–1–septate ascospores with paraphyses that are unbranched and aseptate, while D. menghaica has smaller apothecia, shorter stipe and slightly hyaline to light yellow at the base with branched paraphyses. Dicephalospora albolutea differs from D. maetaengica in having yellowish-white apothecia, longer asci (140–156 × 9.5–10.5 µm vs. 81–111 × 5.6–10.7 µm) and ascospores (26–31 × 3.8–5 µm vs. 14.5–21.8 × 2.4–4.2 µm) with aseptate paraphyses and ascospores (Zheng and Zhuang 2019). Therefore, D. maetaengica is introduced here as a new species.

Dicephalospora menghaica L. Luo & K.D. Hyde, sp. nov.

Index Fungorum: IF902891
Facesoffungi Number: FoF16775
Fig. 4

Etymology.

The epithet “menghaica” refers to the collection site, Menghai County, where the holotype specimen was collected.

Holotype.

HKAS135692.

Description.

Saprobic on a dead leaf. Sexual morph: Apothecia 1–1.6 mm diam. when dry, arising solitary, uniseriate on petiole of leaves. Stipe 0.6–1 mm height and light brown to brown at the base. Receptacle orange and cupulate. Margins smooth and yellow when fresh. Disc slightly sunken and light yellow. Ectal excipulum 21–37 µm (x̄ = 28 µm, n = 60), multi-layered, thin-walled, with hyaline to light yellow cells of textura porrecta. Medullary excipulum 24–50 µm (x̄ = 41 µm, n = 60), composed of thin-walled, hyaline to yellowish, gelatinized cells of textura intricata to globulosa, small cells condensed. Hymenium 83–166 µm (x̄ = 122 µm, n = 60), hyaline to yellowish and inner mixed with asci and paraphyses. Paraphyses 0.8–2.1 µm wide (x̄ = 1.2 µm, n = 60), at the terminal cell, filiform, numerous, and have lengths exceeding the asci, smooth, branched, aseptate, slightly swollen, small, and globose at the apex. Asci (79–)82–125(130–) × 6.5–11.6 µm (x̄ = 101 × 9.4 µm, n = 50), 8-spored, unitunicate, cylindrical, and clavate, with the amyloid (J+) having a rounded apex, tapered, long stipitate base. Ascospores (16–)18–24(26–) × 2–3.7 µm (x̄ = 22 × 2.9 µm, n = 60), uniseriate to biseriate, fusoid-clavate with rounded ends, 7-septate, narrowed ends capped with pigments. Asexual morph: Not observed.

Figure 4. 

Dicephalospora menghaica (HKAS 135692, holotype) a fresh apothecia growing on a dead wood b, c dried apothecia d a vertical section of the apothecium e, f excipulum g paraphyses h paraphyses and part of asci i–m asci (j–m asci in Meltzer’s reagent) n–q ascospores. Scale bars: 3 mm (a); 1 mm (b, c); 100 µm (d); 50 µm (e–m); 20 µm (n–q).

Material examined.

China • Yunnan Province, Xishuangbanna City, Menghai County, altitude 1595 m, on the decayed unidentified leaves, 8 September 2022, Le Luo, ly958 (HKAS 135692, holotype); • ibid., ly972 (HKAS 135690, paratype).

Notes.

Dicephalospora menghaica clustered with D. maetaengica by 100% MLBS and 1.00 BIPP support (Fig. 1), and clustered sister to D. albolutea with 98% MLBS and 1.00 BIPP support (Fig. 1). Dicephalospora menghaica differs from D. albolutea by having smaller (1–1.6 mm vs. 1–2.5 mm), light brown apothecia, slightly hyaline to light yellow at the base with branched, aseptate paraphyses, shorter asci (82–125 × 6.5–11.6 µm vs. 140–156 × 9.5–10.5 µm) and smaller ascospores (18–24 × 2–3.7 µm vs. 26–31 × 3.8–5 µm), whereas D. albolutea has cream to yellowish-white apothecia, with unbranched, septate paraphyses (Zheng and Zhuang 2019). Dicephalospora menghaica also differs morphologically from D. maetaengica as discussed previously under the notes of D. maetaengica. Therefore, based on the morphological and phylogenetic analysis, D. menghaica is introduced here as a new species.

Discussion

The members of Leotiomycetes are characterized by a diverse array of species with various ecological roles, including plant pathogens, endophytes, and saprobes (O’Brien et al. 2005; Sieber 2007; Baral 2016). They occur in aquatic to terrestrial ecosystems and play significant roles in decomposition and nutrient cycling (Quandt and Haelewaters 2021). In China, the diversity of Leotiomycetes is substantial due to the varied climates and ecosystems (Li et al. 2022; Su et al. 2023; Luo et al. 2024; Guo et al. 2024; Zhang et al. 2024). Ongoing research continues to uncover new species and understand their roles in ecosystem functioning, highlighting the importance of preserving fungal diversity for ecological health and agricultural sustainability (Li et al. 2022; Su et al. 2023; Luo et al. 2024).

The identification of Dicephalospora species has traditionally relied on morphological features, such as the color of the apothecia, anatomical structures, and the characteristics of asci and ascospores (Zheng and Zhuang 2019; Phutthacharoen et al. 2022; Luo et al. 2024). However, morphology-based identification became challenging due to the morphological variability observed even within the same species collected from the same geographical locations (Phutthacharoen et al. 2022). Morphological variation within a single fungal species can be influenced by genetic diversity, environmental conditions, transposable elements, and symbiotic relationships (Taylor et al. 2017; Senanayake et al. 2020). Similarly, apothecia and ascospores showed morphological diversity in Dicephalospora rufocornea isolates reported from the same province in Thailand (Phutthacharoen et al. 2022). This variability underscores the difficulty of relying solely on morphological characteristics for species identification and emphasizes the importance of integrating morphological data with molecular phylogenetics for accurate identification and classification. In this study, we updated the dichotomous key for the identification of Dicephalospora species.

DNA sequence data, particularly ITS and LSU gene sequences, play a crucial role in the delineation of fungal species (Hibbett et al. 2016; Jeewon and Hyde 2016). Our phylogenetic analyses based on these sequences provide robust support for the differentiation of the new species within Dicephalospora. The two new species, D. menghaica and D. maetaengica, are characterized by apothecial characters, with paraphyses that are either unbranched or branched. The phylogenetic relationships among Dicephalospora species in our study concurred with those reported by Phutthacharoen et al. (2022), who identified one main cluster and another subclade comprising D. chrysotricha isolates closer to the Hymenoscyphus clade. This discrepancy highlights the need for further research and more extensive sampling to clarify these relationships.

This study expands our understanding of Dicephalospora, particularly through the discovery and description of three new species from Xishuangbanna, Yunnan Province, China, and northern Thailand. Dicephalospora species are primarily saprobic, decomposing organic matter such as rotten wood, twigs, and leaf petioles. This saprobic activity plays a crucial role in nutrient cycling within their ecosystems. The tropical monsoon climate of Xishuangbanna, with its high humidity, provides a unique habitat for fungi, exemplified by the discovery of these species on wet decaying wood (Luo et al. 2024). This finding contrasts with previous observations that Dicephalospora were typically found in highly humid and cold areas, suggesting a broader ecological range and adaptability to various climatic conditions (Phutthacharoen et al. 2022). This study expands the Dicephalospora species up to 22 species, of which 16 were reported from China, four from Thailand (D. chiangraiensis, D. irregularis, D. inthanonensis, D. maetaengica), one from New Zealand (D. chrysotricha), and one from Australia (D. sagerae) (Spooner 1987; Zhuang et al. 2016; Zheng and Zhuang 2019; Phutthacharoen et al. 2022; Luo et al. 2024). China appears to harbor a high diversity of Dicephalospora species that are yet to be discovered.

This study contributes to the taxonomy and phylogeny of Dicephalospora by describing new species and clarifying their relationships within the Helotiaceae. In addition, a dichotomous key to the species in Dicephalospora is presented. Future studies should aim to include a wider range of genera and utilize additional genetic markers to further resolve the phylogenetic position of Dicephalospora and related taxa.

A dichotomous key to the species of Dicephalospora

1 Receptacle surface with hairs D. chrysotricha
– Receptacle surface without hairs 2
2 Sessile apothecia 3
– Stipitate apothecia 5
3 Asci J+ D. calochroa
– Asci J- 4
4 Disc concave with unbranched paraphyses D. sessilis
– Disc slightly convex with branched paraphyses D. irregularis
5 Margin dentate D. dentata
– Margin not dentate 6
6 Disc cream to yellowish, white apothecia D. albolutea
– Disc and apothecia concolorous 7
7 Paraphyses with dark pigment contents D. phaeoparaphysis
– Paraphyses without dark pigment contents 8
8 Asci J- 9
– Asci J+ 11
9 Stipe base dark D. pinglongshanica
– Stipe base not dark 10
10 Apothecia orange when fresh, light brown when dry D. tengyueica
– Apothecia yellow to orange when fresh and dry D. xishuangbannaensis
11 Ascospore cap mucilaginous 12
– Ascospore cap non-mucilaginous 13
12 Ascospore lemon-shaped, 9−12.7 µm wide D. damingshanica
– Ascospore fusoid D. rufocornea
13 Ascospores constricted in the middle D. contracta
– Ascospores not constricted in the middle 14
14 Ascospores elliptical-subfusoid D. shennongjiana
– Ascospores fusoid 15
15 Disc convex D. inthanonensis
– Disc flat 16
16 Paraphyses septate 17
– Paraphyses aseptate 18
17 Ascospores multiseriate, ectal excipulum globose at the tips D. huangshanica
– Ascospores biseriate, 16.5−25.3 × 3.3−3.5 µm D. yunnanica
18 Paraphyses branched D. menghaica
– Paraphyses unbranched 19
19 Ascospore aseptate D. aurantiaca
– Ascospore septate 20
20 Stipe and apothecia concolorous D. maetaengica
– Stipe and apothecia not concolorous D. chiangraiensis

Acknowledgements

Le luo thanks Dr Shaun Pennycook for his valuable assistance with the Latin binomial nomenclature.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This work was supported by the Second Tibetan Plateau Scientific Expedition and Research (STEP) Program (Grant No. 2019QZKK0503). This work also was supported by the Chinese Research Fund (project no E1644111K1) entitled “Flexible introduction of high-level expert program, Kunming Institute of Botany, Chinese Academy of Sciences’’. The authors extend their appreciation to the Researchers Supporting Project number (RSP2025R114), King Saud University, Riyadh, Saudi Arabia.

Author contributions

Supervision: KWTC, KK. Writing - original draft: LL, FAO. Writing - review and editing: VT, QZ, KDH.

Author ORCIDs

Le Luo https://orcid.org/0009-0006-7213-0498

Kandawatte Wedaralalage Thilini Chethana https://orcid.org/0000-0002-5816-9269

Qi Zhao https://orcid.org/0000-0001-8169-0573

Vinodhini Thiyagaraja https://orcid.org/0000-0002-8091-4579

Kitiphong Khongphinitbunjong https://orcid.org/0000-0002-4548-2790

Fatimah Al-Otibi https://orcid.org/0000-0003-3629-5755

Kevin D. Hyde https://orcid.org/0000-0002-2191-0762

Data availability

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

References

  • Baral HO (2016) Inoperculate discomycetes. In: Jaklitsch W, Baral HO, Lücking R, Lumbsch HT, Frey W (Eds) Syllabus of plant families: A. Engler’s syllabus der Pflanzenfamilien part 1/2. Borntraeger, Stuttgart, 157–205.
  • Baral HO, De Sloover J, Huhtinen S, Laukka T, Stenroos S (2009) An emendation of the genus Hyaloscypha to include Fuscoscypha (Hyaloscyphaceae, Helotiales, Ascomycotina). Karstenia 49(1): 1–17. https://doi.org/10.29203/ka.2009.430
  • Bills GF, Platas G, Peláez F, Masurekar P (1999) Reclassification of a pneumocandin-producing anamorph, Glarea lozoyensis gen. et. sp. nov., previously identified as Zalerion arboricola. Mycological Research 103(2): 179–192. https://doi.org/10.1017/S095375629800687X
  • Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T (2009) trimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics (Oxford, England) 25(15): 1972–1973. https://doi.org/10.1093/bioinformatics/btp348
  • Chaiwan N, Gomdola D, Wang S, Monkai J, Tibpromma S, Doilom M, Wanasinghe D, Mortimer P, Lumyong S, Hyde KD (2021) An online database providing updated information of microfungi in the Greater Mekong Subregion. Mycosphere 12(1): 1513–1526. https://doi.org/10.5943/mycosphere/12/1/19
  • Chethana KWT, Manawasinghe S, Hurdeal VG, Bhunjun CS, Appadoo MA, Gentekaki E, Raspé O, Promputtha I, Hyde KD (2021) What are fungal species and how to delineate them? Fungal Diversity 109(1): 1–25. https://doi.org/10.1007/s13225-021-00483-9
  • Ekanayaka AH, Hyde KD, Gentekaki EG, McKenzie EHC, Zhao Q, Bulgakov TS, Camporesi E (2019) Preliminary classification of Leotiomycetes. Mycosphere 10(1): 310–489. https://doi.org/10.5943/mycosphere/10/1/7
  • Gross A, Hosoya T, Queloz V (2014) Population structure of the invasive forest pathogen Hymenoscyphus pseudoalbidus. Molecular Ecology 23(12): 2943–2960. https://doi.org/10.1111/mec.12792
  • Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology 59(3): 307–321. https://doi.org/10.1093/sysbio/syq010
  • Guo MJ, Zhuo L, Wang SJ, Sui XN, Zhou H, Cai SR, Luo JT, Lei RH, Shen XY, Piepenbring M, Hou CL (2024) Hyperdiverse Rhytismatales on twigs of Rhododendron spp. Mycosphere 15(1): 764–880. https://doi.org/10.5943/mycosphere/15/1/6
  • Hall T (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.
  • Han JG, Hosoya T, Sung GH, Shin HD (2014) Phylogenetic reassessment of Hyaloscyphaceae sensu lato (Helotiales, Leotiomycetes) based on multi-gene analyses. Fungal Biology 118(2): 150–167. https://doi.org/10.1016/j.funbio.2013.11.004
  • Hibbett D, Abarenkov K, Kõljalg U, Öpik M, Chai B, Cole J, Wang Q, Crous P, Robert V, Helgason T, Herr JR, Kirk P, Lueschow S, Donnell KO, Nilsson RH, Oono R, Schoch C, Smyth C, Walker DM, Porras-Alfaro A, Taylor JW, Geiser DM (2016) Sequence-based classification and identification of Fungi. Mycologia 108: 1049–1068.
  • Hoang DT, Chernomor O, Von Haeseler A, Minh BQ, Vinh LS (2018) UFBoot2: Improving the ultrafast bootstrap approximation. Molecular Biology and Evolution 35(2): 518–522. https://doi.org/10.1093/molbev/msx281
  • Hosoya T, Hamano K, Sugano M, Ogura Y, Hatano E, Hamada T (1999) Discovery of dicephalosterol, a new testosterone 5α-reductase inhibitor, and some new mycological aspects of its producer, Dicephalospora rufocornea (Sclerotiniaceae, Discomycetes). Mycoscience 40(6): 525–529. https://doi.org/10.1007/BF02461030
  • Jayasiri SC, Hyde KD, Ariyawansa HA, Bhat J, Buyck B, Cai L, Dai YC, Abd-Elsalam KA, Ertz D, Hidayat I, Jeewon R, Jones EBG, Bahkali AH, Karunarathna SC, Liu JK, Luangsa-ard JJ, Lumbsch HT, Maharachchikumbura SSN, McKenzie EHC, Moncalvo J-M, Ghobad-Nejhad M, Nilsson H, Pang K-L, Pereira OL, Phillips AJL, Raspé O, Rollins AW, Romero AI, Etayo J, Selçuk F, Stephenson SL, Suetrong S, Taylor JE, Tsui CKM, Vizzini A, Abdel-Wahab MA, Wen TC, Boonmee S, Dai DQ, Daranagama DA, Dissanayake AJ, Ekanayaka AH, Fryar SC, Hongsanan S, Jayawardena RS, Li WJ, Perera RH, Phookamsak R, de Silva NI, Thambugala KM, Tian Q, Wijayawardene NN, Zhao RL, Zhao Q, Kang JC, Promputtha I (2015) The Faces of Fungi database: Fungal names linked with morphology, phylogeny and human impacts. Fungal Diversity 74(1): 3–18. https://doi.org/10.1007/s13225-015-0351-8
  • Jeewon R, Hyde KD (2016) Establishing species boundaries and new taxa among fungi: Recommendations to resolve taxonomic ambiguities. Mycosphere 7(11): 1669–1677. https://doi.org/10.5943/mycosphere/7/11/4
  • Johnson M, Zaretskaya I, Raytselis Y, Merezhuk Y, McGinnis S, Madden TL (2008) NCBI BLAST: A better web interface. Nucleic Acids Research 36(Web Server): W5–W9. https://doi.org/10.1093/nar/gkn201
  • Johnston PR, Park D (2013) The phylogenetic position of Lanzia berggrenii and its sister species. Junwu Xuebao 32: 366–385.
  • Johnston PR, Quijada L, Smith CA, Baral H-O, Hosoya T, Baschien C, Pärtel K, Zhuang WY, Haelewaters D, Park D, Carl S, López-Giráldez F, Wang Z, Townsend JP (2019) A multigene phylogeny toward a new phylogenetic classification of Leotiomycetes. IMA Fungus 10(1): 1–22. https://doi.org/10.1186/s43008-019-0002-x
  • Kalyaanamoorthy S, Minh BQ, Wong TK, Von Haeseler A, Jermiin LS (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. https://doi.org/10.1038/nmeth.4285
  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772–780. https://doi.org/10.1093/molbev/mst010
  • Korf RP (1973) Discomycetes and tuberales. The fungi, 249–319.
  • Li CJY, Chethana KWT, Lu ZY, Zhao Q (2022) Two novel species of Lachnaceae (Helotiales, Leotiomycetes) from southwestern China. Current Research in Environmental & Applied Mycology (Journal of Fungal Biology) 12: 333–345. https://doi.org/10.5943/cream/12/1/20
  • Li CJY, Chethana KWT, Eungwanichayapant PD, Zhou DQ, Zhao Q (2024a) Additional four species of Tatraea (Leotiomycetes, Helotiales) in Yunnan Province, China. MycoKeys 102: 127–154. https://doi.org/10.3897/mycokeys.102.112565
  • Li CJY, Chethana KWT, Eungwanichayapant PD, Luo L, Yang ZL, Dong WJ, Guo YY, Liu CH, Alotibi F, Hyde KD, Zhao Q (2024b) Longistipes gen. nov. and four novel species of Hyphodiscaceae along with six new collections of Leotiomycetes in Yunnan Province, China. Mycosphere 15(1): 4744–4787. https://doi.org/10.5943/mycosphere/15/1/20
  • Luo L, Wei D, Zhao Q, Chethana KWT (2024) Unveiling the diversity: A novel species of Dicephalospora (Helotiaceae, Helotiales) discovered in China. Phytotaxa 652(1): 59–68. https://doi.org/10.11646/phytotaxa.652.1.5
  • Luo L, Chethana KWT, Zhao Q, Thiyagaraja V, Al-Otibi F, Hyde KD (2025) Unveiling fungal diversity: a novel species of Proliferodiscus (Lachnaceae, Helotiales) discovered in China. Phytotaxa 682(3): 227–239. https://doi.org/10.11646/phytotaxa.682.3.3
  • Nguyen L-T, Schmidt HA, Von Haeseler A, Minh BQ (2015) IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268–274. https://doi.org/10.1093/molbev/msu300
  • O’Brien HE, Parrent JL, Jackson JA, Moncalvo JM, Vilgalys R (2005) Fungal community analysis by large-scale sequencing of environmental samples. Applied and Environmental Microbiology 71(9): 5544–5550. https://doi.org/10.1128/AEM.71.9.5544-5550.2005
  • Peláez F, Collado J, Platas G, Overy DP, Martín J, Vicente F, González del Val A, Basilio A, De la Cruz M, Tormo JR, Fillola A, Arenal F, Villareal M, Rubio V, Baral HO, Galán R, Bills GF (2011) Phylogeny and intercontinental distribution of the pneumocandin-producing anamorphic fungus Glarea lozoyensis. Mycology 2(1): 1–17. https://doi.org/10.1080/21501203.2010.544334
  • Phutthacharoen K, Chethana KWT, Lestari AS, Stadler M, Hyde KD (2022) Three new species of Dicephalospora (Helotiaceae, Helotiales) from Thailand. Diversity 14(8): 645. https://doi.org/10.3390/d14080645
  • Rämä T, Mathiassen GH, Kauserud H (2014) Marine fungi new to Norway, with an outlook to the overall diversity. Agarica : Mykologisk Tidsskrift 35: 35–47.
  • Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • Senanayake IC, Rathnayaka AR, Marasinghe DS, Calabon MS, Gentekaki E, Lee HB, Hurdeal VG, Pem D, Dissanayake LS, Wijesinghe SN, Bundhun D, Nguyen TT, Goonasekara ID, Abeywickrama PD, Bhunjun CS, Jayawardena RS, Wanasinghe DN, Jeewon R, Bhat DJ, Xiang MM (2020) Morphological approaches in studying fungi: collection, examination, isolation, sporulation and preservation. Mycosphere 11(1): 2678–2754. https://doi.org/10.5943/mycosphere/11/1/20
  • Spatafora JW, Sung G-H, Johnson D, Hesse C, O’Rourke B, Serdani M, Spotts R, Lutzoni F, Hofstetter V, Miadlikowska J, Reeb V, Gueidan C, Fraker E, Lumbsch T, Lücking R, Schmitt I, Hosaka K, Aptroot A, Roux C, Miller AN, Geiser DM, Hafellner J, Hestmark G, Arnold AE, Büdel B, Rauhut A, Hewitt D, Untereiner WA, Cole MS, Scheidegger C, Schultz M, Sipman H, Schoch CL (2006) A five-gene phylogeny of Pezizomycotina. Mycologia 98(6): 1018–1028. https://doi.org/10.1080/15572536.2006.11832630
  • Spooner BM (1987) Helotiales. of Australasia: Geoglossaceae, Orbiliaceae, Sclerotiniaceae, Hyaloscyphaceae. Bibliotheca Mycologica 116: 470–474.
  • Stenroos S, Laukka T, Huhtinen S, Döbbeler P, Myllys L, Syrjänen K, Hyvönen J (2010) Multiple origins of symbioses between ascomycetes and bryophytes suggested by a five-gene phylogeny. Cladistics 26(3): 281–300. https://doi.org/10.1111/j.1096-0031.2009.00284.x
  • Su HL, Chethana KWT, Zeng M, Zhao Q (2023) Two new species of Erioscyphella (Lachnaceae) from southwestern. Current Research in Environmental & Applied Mycology 13(1): 16–33. https://doi.org/10.5943/cream/13/1/2 [Journal of Fungal Biology]
  • Tochihara Y, Hosoya T (2022) Examination of the generic concept and species boundaries of the genus Erioscyphella (Lachnaceae, Helotiales, Ascomycota) with the proposal of new species and new combinations based on the Japanese materials. MycoKeys 87: 1–52. https://doi.org/10.3897/mycokeys.87.73082
  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172(8): 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
  • Vu D, Groenewald M, De Vries M, Gehrmann T, Stielow B, Eberhardt U, Al-Hatmi A, Groenewald JZ, Cardinali G, Houbraken J, Boekhout T, Crous PW, Robert V, Verkley GJM (2019) Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation. Studies in Mycology 92(1): 135–154. https://doi.org/10.1016/j.simyco.2018.05.001
  • Wang Z, Binder M, Hibbett D (2005) Life history and systematics of the aquatic discomycete Mitrula (Helotiales, Ascomycota) based on cultural, morphological, and molecular studies. American Journal of Botany 92(9): 1565–1574. https://doi.org/10.3732/ajb.92.9.1565
  • Wijayawardene NN, Hyde KD, Dai DQ, Sánchez-García M, Goto BT, Saxena RK, Erdoğdu M, Selçuk F, Rajeshkumar KC, Aptroot A, Błaszkowski J, Boonyuen N, da Silva GA, de Souza FA, Dong W, Ertz D, Haelewaters D, Jones EBG, Karunarathna SC, Kirk PM, Kukwa M, Kumla J, Leontyev DV, Lumbsch HT, Maharachchikumbura SSN, Marguno F, Martínez-Rodríguez P, Mešić A, Monteiro JS, Oehl F, Pawłowska J, Pem D, Pfliegler WP, Phillips AJL, Pošta A, He MQ, Li JX, Raza M, Sruthi OP, Suetrong S, Suwannarach N, Tedersoo L, Thiyagaraja V, Tibpromma S, Tkalčec Z, Tokarev YS, Wanasinghe DN, Wijesundara DSA, Wimalaseana SDMK, Madrid H, Zhang GQ, Gao Y, Sánchez-Castro I, Tang LZ, Stadler M, Yurkov A, Thines M (2022) Outline of Fungi and fungus-like taxa–2021. Mycosphere 13: 53–453. https://doi.org/10.5943/mycosphere/13/1/2
  • Yokoya K, Postel S, Fang R, Sarasan V (2017) Endophytic fungal diversity of Fragaria vesca, a crop wild relative of strawberry, along environmental gradients within a small geographical area. PeerJ 5: e2860. https://doi.org/10.7717/peerj.2860
  • Zhao YJ (2014) Taxonomic study of Lambertella (Rutstroemiaceae, Helotiales) and allied substratal stroma forming fungi from Japan. University of Tsukuba, Tsukuba, Japan.
  • Zhao YJ, Hosaka K, Hosoya T (2016) Taxonomic re-evaluation of the genus Lambertella (Rutstroemiaceae, Helotiales) and allied stroma-forming fungi. Mycological Progress 15(12): 1215–1228. https://doi.org/10.1007/s11557-016-1225-5
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