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. 2024 Feb 9;102:83–105. doi: 10.3897/mycokeys.102.115452

Identification of two new species and a new host record of Distoseptispora (Distoseptisporaceae, Distoseptisporales, Sordariomycetes) from terrestrial and freshwater habitats in Southern China

Xue-Mei Chen 1,2, Xia Tang 3,4, Jian Ma 1,4, Ning-Guo Liu 1, Saowaluck Tibpromma 2, Samantha C Karunarathna 2,5, Yuan-Pin Xiao 1, Yong-Zhong Lu 1,3,✉
PMCID: PMC10873807  PMID: 38370857

Abstract

During our investigation of saprophytic fungi in Guizhou and Hainan provinces, China, three hyphomycetes were collected from terrestrial and freshwater habitats. Based on morphological characteristics and phylogenetic analyses of combined ITS, LSU, tef1-α, and rpb2 sequence data, two new species are introduced: Distoseptisporahainanensis and D.lanceolatispora. Additionally, one known species, D.tectonae, previously unreported from Edgeworthiachrysantha, is newly reported. Detailed descriptions, illustrations, and a phylogenetic tree to show the two new species and the new host record of Distoseptispora are provided. In addition, a checklist of Distoseptispora species with their locations, lifestyles, habitats, and hosts is provided.

Key words: 2 new taxa, asexual morph, phylogeny, taxonomy

Introduction

Distoseptispora K.D. Hyde, McKenzie & Maharachch. was introduced by Su et al. (2016) with D.fluminicola McKenzie, Hong Y. Su, Z.L. Luo & K.D. Hyde, as the type species. Most Distoseptispora species are reported as saprophytes, typically found on decaying wood in terrestrial and freshwater habitats (Hyde et al. 2016, 2019; Su et al. 2016; Xia et al. 2017; Yang et al. 2018; Crous et al. 2019; Luo et al. 2019). The initial descriptions of Distoseptispora are derived from its asexual morphology (Hyde et al. 2016, 2019, 2020; Su et al. 2016; Yang et al. 2018, 2021; Luo et al. 2019; Sun et al. 2020). The first description of a sexual morph of Distoseptispora was described by Yang et al. (2021). Recently, Konta et al. (2023) identified the second sexual species on dead leaves of Licualaglabra, and provided detailed explanations, enhancing our understanding of Distoseptispora sexual morphology. This sexual morph is characterized by solitary or gregarious, immersed to semi-immersed, subglobose to ellipsoidal, perithecial, dark brown ascomata with a short neck; 8-spored, cylindrical, short pedicellate asci with non-amyloid apical annuli; and fusiform, 0–3-septate, hyaline ascospores with mucilaginous sheaths (Yang et al. 2021; Konta et al. 2023). The asexual morph of Distoseptispora was recently expanded upon by Yang et al. (2021), incorporating macronematous, mononematous, solitary or fasciculate conidiophores, blastic, terminal, percurrent, cylindrical conidiogenous cells; and acrogenous, solitary, obclavate, ellipsoidal, obovoid or fusiform, rostrate or not, euseptate, distoseptate or rarely muriform conidia with or without a septal pore and mucilaginous sheath.

Distoseptispora has been found on various hosts viz. Tectona, Pandanus, bamboo, Clematis, Carex, Dipterocarpus, Licualaglabra, Cocosnucifera, Phragmitesaustralis, Thysanolaenamaxima, Platanusorientalis, and decaying wood and grasses (Shoemaker and White 1985; McKenzie 1995; Hyde et al. 2016, 2019, 2021, 2023; Su et al. 2016; Tibpromma et al. 2018; Crous et al. 2019; Phookamsak et al. 2019; Phukhamsakda et al. 2020, 2022; Sun et al. 2020; Zhai et al. 2022; Afshari et al. 2023; Hu et al. 2023; Konta et al. 2023). Most Distoseptispora species have been described in Asia, mainly in China, Thailand, and Malaysia, and only a few have been described in Europe (Shoemaker and White 1985; McKenzie 1995; Phookamsak et al. 2019; Ma et al. 2022; Zhai et al. 2022; Zhang et al. 2022; Konta et al. 2023). Distoseptispora comprises 74 accepted species in Index Fungorum (2024), but there is an ambiguity in the taxonomic status of D.submersa Z.L. Luo, K.D. Luo et al. (2019) stated that D.submersa is phylogenetically closely related to D.tectonae, and there are only minor size differences in conidiophores and conidia between D.tectonae and D.submersa. Dong et al. (2021) synonymized D.submersa under D.tectonae, thus, Distoseptispora comprises 73 accepted saprobic species, of which 44 were from freshwater habitats, 29 from terrestrial habitats, and five from both terrestrial and freshwater environments (Hyde et al. 2016, 2019; Luo et al. 2019; Monkai et al. 2020; Yang et al. 2021; Ma et al. 2022; Zhang et al. 2022; Afshari et al. 2023; Hu et al. 2023; Konta et al. 2023; Liu et al. 2023).

In this study, three fresh hyphomycetous fungal collections were encountered during a microfungal investigation in Hainan and Guizhou provinces. Based on multi-gene phylogeny and morphological comparison, two new species, Distoseptisporahainanensis and D.lanceolatispora are introduced. In addition, a new host record of D.tectonae from Edgeworthiachrysantha is also reported.

Materials and methods

Sample collection, isolation, and morphological study

Fresh specimens were collected from Hainan and Guizhou provinces in China. Fungal colonies were mounted on a slide with distilled water and were observed and examined using a stereomicroscope (SMZ 745, Nikon, Tokyo, Japan). Micro-morphological characteristics were captured with a Nikon EOS 90D digital camera combined with an ECLIPSE Ni-U compound microscope (Nikon, Tokyo, Japan). The sizes of the fungal structures were measured using the Tarosoft (R) Image Frame Work program (IFW 0.97 version), and the photo plates were processed with Adobe Photoshop CC 2019 (Adobe Systems, San Jose, CA, USA).

Single spore isolations were carried out following the methods described in Senanayake et al. (2020). Germinated conidia were transferred to fresh potato dextrose agar (PDA) plates and incubated at 25–27 °C for four weeks. Culture characteristics, including color, shape, and size, were recorded. Herbarium specimens were deposited in the herbarium of the Guizhou Academy of Agriculture Sciences (GZAAS), Guiyang, China, and the living cultures were deposited at the Guizhou Culture Collection, China (GZCC). Faces of Fungi and Index Fungorum numbers were obtained following the protocols outlined by Jayasiri et al. (2015) and Index Fungorum (2024), respectively.

DNA extraction, PCR amplification, and sequencing

Fresh mycelia were scraped from cultures that were incubated at 25–27 °C for 28 days. Fungal genomic DNA was extracted using the Biospin Fungus Genomic DNA Extraction Kit (BioFlux, Shanghai, China), following the manufacturer’s instructions. Four gene regions: internal transcribed spacer (ITS), large subunit ribosomal DNA (LSU), translation elongation factor 1-alpha (tef1-α), and RNA polymerase II second largest subunit (rpb2) were selected. The primers used in this study for each gene region were as follows: ITS4 and ITS5 for ITS (White et al. 1990), LR0R and LR5 for LSU (Vilgalys and Hester 1990; Cubeta et al. 1991), EF1-983F and EF1-2218R for tef1-α (Rehner and Samuels 1994), and rpb2 with fRPB2-5F and fRPB2-7cR (Liu et al. 1999).

Polymerase chain reaction (PCR) amplifications were carried out in a 50 µL reaction volume containing 44 μL of 1.1 × T3 Super PCR Mix (TsingKe Biotech, Chongqing, China), 2 µL of DNA template, and 2 µL of each forward and reverse primer. The amplification condition for LSU and ITS consisted of initial denaturation at 94 °C for 3 min, followed by 35 cycles of 45 s at 94 °C, 50 s at 56 °C, and 1 min at 72 °C, and a final extension period of 10 min at 72 °C. The amplification condition for the tef1-α gene consisted of initial denaturation at 94 °C for 3 min, followed by 30 cycles of 30 s at 94 °C, 50 s at 56 °C, and 1 min at 72 °C, a final extension period of 10 min at 72 °C. The amplification condition for the rpb2 gene consisted of initial denaturation at 95 °C for 5 min, followed by 35 cycles of 15 s at 95 °C, 50 s at 56 °C, and 1 min at 72 °C, a final extension period of 10 min at 72 °C. The quality of PCR amplification products was examined with 1% agarose electrophoresis gels stained with ethidium bromide, and the PCR products were sent to TsingKe Biotech, Chongqing, China for purification and sequencing.

Phylogenetic analyses

The raw sequences were initially checked with BioEdit v 7.0.5.3 (Hall 1999). Forward and reverse sequences were assembled using SeqMan v. 7.0.0 (DNASTAR, Madison, WI, USA). Sequence data (LSU, ITS, tef1-α, and rpb2) for Distoseptispora were downloaded from GenBank based on the blast results and recent publications (Table 1). Each individual gene dataset was aligned using the online program MAFFT version 7 with the “auto” option (Hall 1999; Katoh and Standley 2013). These alignments were visually inspected and manually improved in BioEdit v 7.0.5.3. Multi-gene alignments were combined by SequenceMatrix (Vaidya et al. 2011). In this study, phylogenetic analyses were performed using maximum likelihood (ML), maximum parsimony (MP), and Bayesian posterior probability (BYPP) methods. The analyses were based on LSU, ITS, tef1-α, and rpb2 combined sequence datasets.

Table 1.

Names, strain numbers, and corresponding GenBank accession numbers of taxa used in this study.

Taxa names Strain GenBank Accessions References
LSU ITS tef1-α rpb2
Aquapteridosporaaquatica MFLUCC 17-2371T MW287767 MW286493 N/A N/A Dong et al. (2021)
Distoseptisporaadscendens HKUCC 10820 DQ408561 N/A N/A DQ435092 Shenoy et al. (2006)
D.amniculi MFLU 17-2129T MZ868761 MZ868770 N/A MZ892982 Yang et al. (2021)
D.appendiculata MFLUCC 18-0259T MN163023 MN163009 MN174866 N/A Luo et al. (2019)
D.aqualignicola KUNCC 21-10729T ON400845 OK341186 OP413480 OP413474 Zhang et al. (2022)
D.aquamyces KUNCC 21-10731T OK341199 OK341187 OP413482 OP413476 Zhang et al. (2022)
D.aquatica MFLUCC 15-0374T KU376268 MF077552 N/A N/A Su et al. (2016)
MFLUCC 18-0646 MK849793 MK828648 N/A N/A Luo et al. (2019)
D.aquisubtropica GZCC 22-0075T ON527941 ON527933 ON533677 ON533685 Ma et al. (2022)
D.atroviridis GZCC 20-0511T MZ868763 MZ868772 MZ892978 MZ892984 Yang et al. (2021)
D.bambusae MFLUCC 20-0091T MT232718 MT232713 MT232880 MT232881 Sun et al. (2020)
MFLUCC 14-0583 MT232717 MT232712 N/A MT232882 Sun et al. (2020)
D.bambusicola GZCC 21-0667T MZ474872 MZ474873 N/A N/A Hyde et al. (2023)
D.bangkokensis MFLUCC 18-0262T MZ518206 MZ518205 N/A N/A Shen et al. (2021)
D.cangshanensis MFLUCC 16-0970T MG979761 MG979754 MG988419 N/A Luo et al. (2018)
D.caricis CPC 36498T MN567632 MN562124 N/A MN556805 Crous et al. (2019)
CPC 36442 N/A MN562125 N/A MN556806 Crous et al. (2019)
D.chinensis GZCC 21-0665T MZ474867 MZ474871 MZ501609 N/A Hyde et al. (2021)
D.clematidis MFLUCC 17-2145T MT214617 MT310661 N/A MT394721 Phukhamsakda et al. (2020)
D.crassispora KUMCC 21-10726T OK341196 OK310698 OP413479 OP413473 Zhang et al. (2022)
D.curvularia KUMCC 21-10725T OK341195 OK310697 OP413478 OP413472 Zhang et al. (2022)
D.cylindricospora DLUCC 1906T OK513523 OK491122 OK524220 N/A Phukhamsakda et al. (2022)
D.dehongensis KUMCC 18-0090T MK079662 MK085061 MK087659 N/A Hyde et al. (2019)
D.dipterocarpi MFLUCC 22-0104T OP600052 OP600053 N/A OP595140 Afshari et al. (2023)
D.effusa GZCC 19-0532T MZ227224 MW133916 N/A N/A Yang et al. (2021)
D.euseptata MFLUCC 20-0154T MW081544 MW081539 N/A MW151860 Li et al. (2021)
MFLU 20-0568 MW081545 MW081540 MW084994 MW084996 Li et al. (2021)
D.fasciculata KUMCC 19-0081T MW287775 MW286501 MW396656 N/A Dong et al. (2021)
D.fluminicola MFLUCC 15-0417T KU376270 MF077553 N/A N/A Su et al. (2016)
D.fusiformis GZCC 20-0512T MZ868764 MZ868773 MZ892979 MZ892985 Yang et al. (2021)
D.gasaensis HJAUP C2034T OQ942891 OQ942896 OQ944455 N/A Hu et al. (2023)
D.guanshanensis HJAUP C1063T OQ942898 OQ942894 OQ944452 OQ944458 Hu et al. (2023)
D.guizhouensis GZCC 21-0666T MZ474869 MZ474868 MZ501610 MZ501611 Hyde et al. (2021)
D.guttulata MFLUCC 16-0183T MF077554 MF077543 MF135651 N/A Yang et al. (2018)
DLUCC B43 MN163016 MN163011 N/A N/A Luo et al. (2019)
D.hainanensis GZCC 22-2047T OR438894 OR427328 OR449122 OR449119 This study
D.hyalina MFLUCC 17-2128T MZ868760 MZ868769 MZ892976 MZ892981 Yang et al. (2021)
D.hydei MFLUCC 20-0481T MT742830 MT734661 N/A MT767128 Monkai et al. (2020)
D.jinghongensis HJAUP C2120T OQ942893 OQ942897 OQ944456 N/A Hu et al. (2023)
D.lancangjiangensis KUN-HKAS 112712T MW879522 MW723055 N/A MW882260 Shen et al. (2021)
D.lanceolatispora GZCC 22-2045T OR43BB95 OR427329 OR449123 OR449120 This study
D.leonensis HKUCC 10822 DQ408566 N/A N/A DQ435089 Shenoy et al. (2006)
D.licualae MFLUCC 14-1163AT ON650675 ON650686 ON734007 N/A Konta et al. (2023)
MFLUCC 14-1163BT ON650676 ON650687 ON734008 N/A Konta et al. (2023)
D.lignicola MFLUCC 18-0198T MK849797 MK828651 N/A N/A Luo et al. (2019)
D.longispora HFJAU 0705T MH555357 MH555359 N/A N/A Song et al. (2020)
D.longnanensis HJAUP C1040T OQ942886 OQ942887 OQ944451 N/A Hu et al. (2023)
D.martinii CGMCC 3.18651T KX033566 KU999975 N/A N/A Xia et al. (2017)
D.meilingensis JAUCC 4727T OK562396 OK562390 OK562408 N/A Zhai et al. (2022)
D.menghaiensis HJAUP C2045T OQ942900 OQ942890 N/A N/A Hu et al. (2023)
HJAUP C2170T OQ942888 OQ942899 OQ944457 OQ944461 Hu et al. (2023)
D.mengsongensis HJAUP C2126T OP78784 OP787876 OP961937 N/A Liu et al. (2023)
D.multiseptata MFLUCC 16-1044 MF077555 MF077544 MF135652 MF135644 Yang et al. (2018)
MFLUCC 15-0609T KX710140 KX710145 MF135659 N/A Hyde et al. (2016)
D.nabanheensis HJAUP C2003T OP787877 OP787873 OP961935 N/A Liu et al. (2023)
D.nanchangensis HJAUP C1074T OQ942895 OQ942889 OQ944454 OQ944460 Hu et al. (2023)
D.neorostrata MFLUCC 18-0376T MN163017 MN163008 N/A N/A Luo et al. (2019)
D.nonrostrata KUNCC 21-10730T OK341198 OK310699 OP413481 OP413475 Zhang et al. (2022)
D.obclavata MFLUCC 18-0329T MN163010 MN163012 N/A N/A Luo et al. (2019)
D.obpyriformis MFLUCC 17-1694T MG979764 N/A MG988422 MG988415 Luo et al. (2018)
DLUCC 0867 MG979765 MG979757 MG988423 MG988416 Luo et al. (2018)
D.pachyconidia KUMCC 21-10724T OK341194 OK310696 OP413477 OP413471 Zhang et al. (2022)
D.palmarum MFLUCC 18-1446T MK079663 MK085062 MK087660 MK087670 Hyde et al. (2019)
D.phangngaensis MFLUCC 16-0857T MF077556 MF077545 MF135653 N/A Yang et al. (2018)
D.phragmiticola GUCC 22-0202T OP749881 OP749888 OP749892 OP752700 Hyde et al. (2023)
D.rayongensis MFLUCC 18-0415T MH457137 MH457172 MH463253 MH463255 Hyde et al. (2020)
MFLUCC 18-0417 MH457138 MH457173 MH463254 MH463256 Hyde et al. (2020)
D.rostrata MFLUCC 16-0969T MG979766 MG979758 MG988424 MG988417 Luo et al. (2018)
DLUCC 0885 MG979767 MG979759 MG988425 N/A Luo et al. (2018)
D.saprophytica MFLUCC 18-1238T MW287780 MW286506 MW396651 MW504069 Dong et al. (2021)
D.septata GZCC 22-0078T ON527947 ON527939 ON533683 ON533690 Ma et al. (2022)
D.sinensis HJAUP C2044T OP787875 OP787878 OP961936 N/A Liu et al. (2023)
D.songkhlaensis MFLUCC 18-1234T MW287755 MW286482 MW396642 N/A Dong et al. (2021)
D.suoluoensis MFLUCC 17-0224T MF077557 MF077546 MF135654 N/A Yang et al. (2018)
MFLUCC 17-1305 MF077558 MF077547 N/A N/A Yang et al. (2018)
D.tectonae MFLUCC 12-0291T KX751713 KX751711 KX751710 KX751708 Hyde et al. (2016)
MFLU 20-0262 MT232719 MT232714 N/A N/A Sun et al. (2020)
MFLUCC 16-0946 MG979768 MG979760 MG988426 MG988418 Dong et al. (2021)
D.tectonae GZCC 22-2046 OR348896 OR427330 OR449124 OR449121 This study
D.tectonigena MFLUCC 12-0292T KX751714 KX751712 N/A KX751709 Hyde et al. (2016)
D.thailandica MFLUCC 16-0270T MH260292 MH275060 MH412767 N/A Tibpromma et al. (2018)
D.thysanolaenae KUN-HKAS 102247T MK064091 MK045851 MK086031 N/A Phukhamsak et al. (2019)
D.tropica GZCC 22-0076T ON527943 ON527935 ON533679 ON533687 Ma et al. (2022)
D.verrucosa GZCC20-0434T MZ868762 MZ868771 MZ892977 MZ892983 Yang et al. (2021)
D.wuzhishanensis GZCC 22-0077T ON527946 ON527938 ON533682 N/A Ma et al. (2022)
D.xishuangbannaensis KUMCC 17-0290T MH260293 MH275061 MH412768 MH412754 Tibpromma et al. (2018)
D.yichunensis HJAUP C1065T OQ942892 OQ942885 OQ944453 OQ944459 Hu et al. (2023)
D.yongxiuensis JAUCC 4725T OK562394 OK562388 OK562406 N/A Zhai et al. (2022)
D.yunjushanensis JAUCC 4723T OK562398 OK562392 OK562410 N/A Zhai et al. (2022)
D.yunnanensis MFLUCC 20-0153T MW081546 MW081541 MW084995 MW151861 Li et al. (2021)
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Note: “T” denotes ex-type strain. Newly generated sequences are indicated in black bold. “N/A”: no data available in GenBank.

The phylogenetic analyses were conducted using the CIPRES Science Gateway V. 3.3. “RAxML-HPC v.8 on XSEDE”, “PAUP on XSEDE”, and “MrBayes on XSEDE (3.2.7a)” were utilized for ML, MP, and BYPP methods, respectively (Huelsenbeck and Ronquist 2001; Swofford 2002; Stamatakis et al. 2008; Miller et al. 2010; Ronquist et al. 2012). For the ML analysis, the GTRGAMMA model of nucleotide evolution was employed, and RAxML rapid bootstrapping with 1,000 bootstrap replicates was obtained (Stamatakis et al. 2008).

The MP analysis employed 1,000 random taxa additions to infer trees. Branches of zero length were collapsed, and all multiple parsimonious trees were saved. The maxtrees value was set to 5,000. For trees generated using different optimal criteria, parsimony score values were determined for tree length (TL), consistency index (CI), retention index (RI), and homoplasy index (HI). To assess clade stability, the bootstrap (BT) method was used with 1,000 iterations, each consisting of 100 trials of random stepwise addition of taxa (Hillis and Bull 1993).

The posterior probabilities (PP) were determined based on Bayesian Markov chain Monte Carlo sampling (Huelsenbeck and Ronquist 2001). The best nucleotide substitution model for each data partition was determined using the program MrModeltest 2.2 (Nylander 2004). The GTR + I + G substitution model with gamma rates and Dirichlet base frequencies was selected for all LSU, ITS, tef1-α, and rpb2 sequences. To calculate the posterior probabilities, four simultaneous Markov chains were run for one million generations, with trees sampled every 100th generation, resulting in a total of 10,000 trees. A burn-in parameter of 0.25 was set, indicating that 75% of the trees were remined during the burn-in phase, and the remaining trees were used for calculating the posterior probabilities in the majority rule consensus tree.

FigTree v. 1.4.4. was used for visualizing the phylogenetic trees, and Adobe Illustrator CC 2019v. 23.1.0 was used to edit trees and figure layout.

Phylogenetic analyses results

This study utilized a combined multi-gene dataset encompassing ITS, LSU, tef1-α, and rpb2 sequences to assess the phylogenetic relationships among Distoseptispora species. The analyses included a total of 90 taxa, designating Aquapteridosporaaquatica X.D. Yu, W. Dong & H. Zhang (MFLUCC 17-2371) as the outgroup taxon. The combined aligned sequence matrix comprised 3,360 characters, including gaps: LSU (1–840 bp), ITS (841–1406 bp), tef1-α (1407–2321 bp), and rpb2 (2322–3360 bp). The ML, MP, and Bayesian trees analyzed exhibited a high degree of similarity in topology and showed no significant conflicts. The RAxML analysis yielded a best-scoring tree (ln = -31666.963504), which is presented in Fig. 1. The matrix encompassed 1572 distinct alignment patterns, with 27.15% constituted by undetermined characters or gaps. The estimated base frequencies were as follows: A = 0.239306, C = 0.265297, G = 0.281926, T = 0.213472; substitution rates AC = 1.429077, AG = 3.512798, AT = 1.204511, CG = 0.845859, CT = 6.948345, GT = 1.000000; gamma distribution shape parameter α = 0.244431. For the MP analysis, 3360 characters remained unchanged, 330 were variable and parsimoniously uninformative, and 1074 were parsimoniously informative. The most parsimonious tree yielded the following values: TL = 5624, CI = 0.400, RI = 0.738, RC = 0.295, HI = 0.600. For BYPP analysis, Bayesian posterior probabilities from MCMC were evaluated with a final average standard deviation of split frequencies of 0.009754.

Figure 1.

Figure 1.

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Phylogenetic tree generated from ML analysis based on a combination of LSU, ITS, tef1-a, and rpb2 sequence data. Bootstrap support values of ML and MP equal to or greater than 75%, and PP value equal to or greater than 0.95 are given near the nodes as ML/PP/MP. The tree is rooted with Aquapteridosporaaquatica (MFLUCC 17-2371). Ex-type strains are indicated by the superscript T. The new collections are in bold red text.

In the phylogenetic analyses (Fig. 1), all our newly identified taxa nested within Distoseptispora, affirming their classification within this genus. Distoseptisporahainanensis (GZCC 22-047) formed a sister clade to D.multiseptata strains (MFLUCC 16-1044 and MFLUCC 15-0609) with 98% ML, 1.00 PP, and 93% MP statistical support. Distoseptisporalanceolatispora (GZCC 22-2045) formed a sister clade to D.neorostrata (MFLUCC 18-0376) with 100% ML, 1.00 PP, and 98% MP statistical support. In addition, our new collection GZCC 22-2046 clustered together with three D.tectonae strains (MFLU 20-0262 and MFLUCC 12-0291) with 98% ML and 0.96 PP statistical support, indicating they represent the same species.

Taxonomy

. Distoseptispora hainanensis

X.M. Chen & Y.Z. Lu sp. nov.

1369CFAC-E4DC-5C03-8878-CEA7116DE0F3

Index Fungorum: IF900953

Facesoffungi Number: FoF14663

Fig. 2

Figure 2.

Figure 2.

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Distoseptisporahainanensis (GZAAS 22-2047, holotype) a, b colonies on substrate c–e conidiophores and conidia f–h conidiogenous cells bearing conidia i, j conidiophores k–q conidia r, s colony on PDA (r from front s from reverse). Scale bars: 50 μm (c, d, f–j, l–q); 30 μm (e, k).

Etymology.

The epithet refers to the location “Hainan Province” where the holotype was collected.

Holotype.

GZAAS 22-2047.

Description.

Saprobic on decaying wood in terrestrial habitat. Sexual morph: Undetermined. Asexual morph: Colonies on natural substrate superficial, effuse, dark brown, and hairy. Mycelium mostly immersed, composed of branched, septate, brown to dark brown, smooth hyphae. Conidiophores 70–130 × 5–8.5 μm (x– = 103 × 7 μm, n = 20), macronematous, mononematous, erect, solitary, straight or slightly flexuous, brown to dark brown, paler towards the apex, cylindrical, 4–6-septate, slightly constricted and darkened at septa, unbranched, thick-walled. Conidiogenous cells 6–13 × 3.5–6.5 μm (x– = 10 × 5 μm, n = 20), holoblastic, monoblastic, integrated, terminal, indeterminate, cylindrical, slightly tapering towards the apex, brown, percurrent. Conidia 44–117 μm × 9–18.5 μm (x– = 90 × 14 μm, n = 20), acrogenous, solitary, obclavate or obpyriform, rostrate, truncate at the base, straight or slightly curved, up to 22-distoseptate, slightly constricted at septa, brown, verrucose.

Culture characteristics.

Colonies grown on PDA circular, dense, fluffy, with raised center and lobate edge, pale gray in the center, grayish brown in the outer ring from the front view, dark brown in the center, and blackish brown in the outer ring from the reverse view.

Material examined.

China, Hainan Province, on unidentified decaying wood, 15 May 2021, Xia Tang, HN02 (GZAAS 22-2047, holotype), ex-type living culture, GZCC 22-2047.

Notes.

Morphologically, Distoseptisporahainanensis is similar to D.effusa L.L. Liu & Z.Y. Liu in having macronematous conidiophores, monoblastic conidiogenous cells, and acrogenous, obclavate, rostrate conidia (Yang et al. 2021). However, conidia of D.hainanensis are up to 22-distoseptate, whereas those of D.effusa are only 4–9-distoseptate. In the phylogenetic analyses, D.hainanensis formed a distinct clade sister to D.multiseptata Jiao Yang & K.D. Hyde with 98% ML, 1 PP, and 93% MP statistical support (Fig. 1). Distoseptisporahainanensis differs from D.multiseptata in having brown, longer conidiophores (70–130 μm vs. 23–65 µm) and obclavate or obpyriform, brown, verrucose, smaller conidia (44–117 μm vs. up to 290 µm) (Hyde et al. 2016). Comparing DNA sequence data, D.hainanensis diverges from D.multiseptata (MFLUCC 15-0609) in the ITS by 21/552 bp (3.8% difference), in the LSU by 1/812 bp (0.01% difference), in tef1-α by 33/912 bp (3.6% difference), and no data is available for rpb2 of D.multiseptata (MFLUCC 15-0609) in GenBank. Hence, the novel species, D.hainanensis, is introduced, following the guidelines of Jeewon and Hyde (2016) and Chethana et al. (2021).

. Distoseptispora lanceolatispora

X.M. Chen & Y.Z. Lu sp. nov.

558A79C2-EC0D-5BBD-BFB7-91B2EE3E304A

Index Fungorum: IF900954

Facesoffungi Number: FoF14664

Fig. 3

Figure 3.

Figure 3.

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Distoseptisporalanceolatispora (GZAAS 22-2045, holotype) a, b colonies on substrate c–e conidiophores and conidia f, g conidiogenous cells bearing conidia h–k conidia l germinated conidium m, n colony on PDA (m from front n from reverse). Scale bars: 50 μm (c–g); 30 μm (h–l).

Etymology.

Referring to the lanceolate conidia.

Holotype.

GZAAS 22-2045.

Description.

Saprobic on submerged decaying wood in freshwater habitat. Sexual morph: Undetermined. Asexual morph: Colonies on substrate effuse, gregarious, hairy, pale brown to brown. Mycelium mostly immersed, composed of septate, yellow-brown to brown, smooth hyphae. Conidiophores 120–190 × 4–8 µm (x– = 155 × 6.5 µm, n = 20), macronematous, mononematous, erect, solitary, straight or slightly flexuous, grayish brown to dark brown, slightly tapering towards the apex, cylindrical, 7–8-septate, unbranched, thick-walled, smooth-walled. Conidiogenous cells 15–27 × 3–5.5 µm (x– = 20.5 × 4.5 µm, n = 20), monoblastic, integrated, terminal, cylindrical, slightly tapering towards the apex, pale brown, percurrent. Conidia 31–90 × 9.5–15 µm (x– = 58.5 × 13 µm, n = 20), acrogenous, solitary, fusiform or lanceolate, rostrate, truncate at the base, straight or slightly curved, 5–13-distoseptate, slightly constricted at septa, olivaceous to olivaceous brown, slightly paler at the apex, verrucous, with or without apical, hyalina appendages.

Culture characteristics.

Colonies grown on PDA circular, dense, flat, dry, gray to dark gray, radially striated, and a ring in the middle of the colonies with an entire edge from the front view, dark brown to black with a circular, gray edge from reverse view, not pigmented.

Material examined.

China, Hainan Province, on submerged decaying wood in a freshwater stream, 23 October 2021, Jian Ma, J13 (GZAAS 22-2045, holotype), ex-type living culture, GZCC 22-2045.

Notes.

Distoseptisporalanceolatispora is morphologically similar to D.leonensis (M.B. Ellis) R. Zhu & H. Zhang. However, compared to D.lanceolatispora, D.leonensis has longer conidiophores (120–190 µm vs. 110–130 µm), longer conidiogenous cells (15–27 µm vs. 5–15 µm), and 5–13-distoseptate, fusiform or lanceolate conidia (Zhang et al. 2022). In the phylogenetic analyses (Fig. 1), D.lanceolatispora forms a unique clade adjacent to D.neorostrata D.F. Bao, Z.L. Luo & H.Y. Su with 100% ML, 1 PP, and 98% MP support. Based on a pairwise nucleotide comparison of ITS and LSU sequences, D.lanceolatispora deviates from D.neorostrata by 39/529 bp (6.8%) for ITS and 14/850 bp (1.6%) for LSU, and there is no data available for tef1-α and rpb2 for D.neorostrata (MFLUCC 18-0376) in GenBank. Hence, we introduce the new species, D.lanceolatispora, based on the criteria established by Jeewon and Hyde (2016) and Chethana et al. (2021).

. Distoseptispora tectonae

Doilom & K.D. Hyde, Fungal Diversity 80: 222 (2016)

E502DE4A-6E52-505B-B0FD-A7463654155E

Index Fungorum: IF552223

Facesoffungi number: FoF01877

Fig. 4

Figure 4.

Figure 4.

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Distoseptisporatectonae (GZAAS 22-2046) a, b colonies on substrate c, d conidiophores and conidia e, f conidiophores g–k conidia l germinated conidium m, n colonies on PDA (m from front n from reverse) Scale bars: 50 μm (c, d, g–l); 20 μm (e, f).

Description.

Saprobic on dead twigs of Edgeworthiachrysantha. Sexual morph: Undetermined. Asexual morph: Colonies on natural substrate abundant, superficial, dark brown, hairy. Conidiophores 35–80 μm × 4–7.5 μm (x– = 58 × 5.5 μm, n = 20), macronematous, mononematous, simple, erect to slightly curved, solitary, pale brown to dark brown, cylindrical, 2–4-septate, slightly constricted at the septa, unbranched, thick-walled. Conidiogenous cells 6–10 μm × 3.5–6.5 μm (x– = 8 × 4.5 μm, n = 20), holoblastic, monoblastic, integrated, terminal, cylindrical, slightly tapering towards the apex, brown to reddish brown, percurrent. Conidia 190–255 μm × 9.5–16 μm (x– = 220 μm × 13 μm, n = 20), 5–16 μm (x– = 13 μm, n = 20) wide at the protruding truncate base; 4.5–8 μm (x– = 6.5 μm, n = 20) wide in the tapering part, acrogenous, solitary, obclavate, elongate, rostrate, straight or curved, tapering towards the apex, 9–39-distoseptate, olivaceous-green when young, dark reddish brown at maturity, verrucose.

Culture characteristics.

Conidia germinating on PDA within 24 h, colonies circular, dense, umbonate, spreading, fluffy. The surface is slightly rough with reddish-gray mycelium, colonies somewhat raised in the middle, and with a filiform edge. The reverse side is dark gray with a circular, pale reddish-gray edge, not pigmented.

Material examined.

China, Guizhou Province, Guiyang City, Guiyang Medicinal Botanical Garden, on dead twigs of Edgeworthiachrysantha, 20 August 2022, Xia Tang, JX30 (GZAAS 22-2046), living culture, GZCC 22-2046.

Known host and distribution.

Tectonagrandis (Thailand, Hyde et al. 2016), on dead stems (Thailand, Sun et al. 2020), on dead, submerged, decaying wood of unidentified plants (China & Thailand, Luo et al. 2019; Dong et al. 2021; Zhang et al. 2022), and dead twig and branch of Edgeworthiachrysantha (China, this study).

Notes.

Distoseptisporatectonae was first isolated from a dead twig of Tectonagrandis in Thailand (Hyde et al. 2016). Since then, this species has been identified in various countries on different substrates and hosts (Hyde et al. 2016; Sun et al. 2020; Dong et al. 2021; Zhang et al. 2022). In the phylogenetic tree (Fig. 1), our new isolate forms a close lineage to D.tectonae (GZCC 22-2046) with statistical support of 98% ML and 0.96 PP. Based on pairwise nucleotide comparisons of ITS, LSU, tef1-α, and rpb2, our new isolate diverges from D.tectonae (MFLUCC 12-0291, ex-type) by 6/554 bp (1%) for ITS, 1/852 bp (0.01%) for LSU, 0/980 bp (0%) for tef1-α, and 2/899 bp (0.2%) for rpb2. In addition, the morphological characteristics of our isolate match well with the holotype description of D.tectonae (Hyde et al. 2016). This study reports a new host record of Distoseptisporatectonae on dead twigs of Edgeworthiachrysantha in China.

Discussion

Distoseptispora is one of the sporidesmium-like taxa and is well-known for its asexual morph, which has considerable morphological variations (Su et al. 2016; Yang et al. 2018, 2021). However, the phylogenetic analyses suggest a lack of correlation between phylogenetic relationships and morphological analyses. For instance, species such as D.appendiculata D.F. Bao, Z.L. Luo & H.Y. Su, D.atroviridis J. Yang & K.D. Hyde, D.caricis Crous, D.fusiformis J. Yang & K.D. Hyde, D.lanceolatispora, D.leonensis, D.neorostrata, D.palmarum S.N. Zhang, K.D. Hyde & J.K. Liu, and D.saprophytica W. Dong, H. Zhang & K.D. Hyde cluster together as a subclade in the phylogenetic tree (see Fig. 1). In contrast, morphological analysis reveals significant differences, especially in the characteristics of conidiophores, conidiogenous cells, and conidia (Crous et al. 2019; Hyde et al. 2019; Luo et al. 2019; Dong et al. 2021; Yang et al. 2021; Zhang et al. 2022). This disparity is common within the genus. We recommend adopting a combination approach using molecular and morphological methods for more effective identification within this genus.

Worth noting, among the various species of Distoseptispora, D.martinii (J.L. Crane & Dumont) J.W. Xia & X.G. Zhang stands out due to its unique morphological characteristics, especially its oblate or subglobose conidia, distinguishing it from other species within Distoseptispora (Xia et al. 2017). The species was initially introduced as Acrodictysmartinii J.L. Crane & Dumont by Crane and Dumont (1975) based on morphological characteristics. Then, it underwent several taxonomic revisions based solely on morphology (Baker et al. 2002; Delgado 2009). Later, Xia et al. (2017) reclassified Acrodictysmartinii as D.martinii based on genetic analysis. However, the morphological traits of D.martinii greatly diverge from typical Distoseptispora features (Crane and Dumont 1975; Xia et al. 2017). Therefore, we suggest additional collections and analysis of D.martinii specimens to ensure the reliability of the provided DNA sequence data.

In recent years, Distoseptispora species have been reported worldwide, such as in China, Hungary, Hawaii, Malaysia, and Thailand (Shoemaker and White 1985; McKenzie 1995; Wu and Zhuang 2005; Zhang et al. 2022). Studies on Distoseptispora have been particularly extensive in China and Thailand (Hyde et al. 2016, 2019, 2020; Su et al. 2016; Yang et al. 2018, 2021; Luo et al. 2019; Sun et al. 2020; Hu et al. 2023). To date, 73 species of Distoseptispora have been documented, of which 55 have been recorded in China (including known species, see Table 2). Our collections further highlight the distribution of the genus in China, and we speculate that the country may harbor a greater diversity of the genus. Thus, future studies are needed to validate this hypothesis.

Table 2.

Distoseptispora species and their locations, lifestyles, habitats, hosts, and corresponding references.

Species Country Habitat Host References
D.adscendens China; Hungary; Hawaii Terrestrial Decaying wood and decaying branches of many woody plant species; Platanusorientalis Shoemaker et al. (1985); McKenzie et al. (1995); Wu et al. (2005); Zhang et al. (2022)
D.amniculi Thailand Freshwater Submerged decaying wood Yang et al. (2021)
D.appendiculata Thailand Freshwater Submerged decaying wood Luo et al. (2019)
D.aqualignicola China Freshwater Submerged decaying wood Zhang et al. (2022)
D.aquamyces China Freshwater Submerged decaying wood Zhang et al. (2022)
D.aquatica China Freshwater Submerged decaying wood Su et al. (2016); Luo et al. (2019); Li et al. (2021)
D.aquisubtropica China Freshwater Submerged decaying wood Ma et al. (2022)
D.atroviridis China Freshwater Submerged decaying wood Yang et al. (2021)
D.bambusae China Terrestrial Decaying bamboo culms Sun et al. (2020)
D.bambusicola China Freshwater Submerged bamboo culms Jayawardena et al. (2022)
D.bangkokensis Thailand Freshwater Submerged decaying wood Shen et al. (2021)
D.cangshanensis China Freshwater Submerged decaying wood Luo et al. (2018)
D.caricis Thailand Terrestrial Leaves of Carex sp. Crous et al. (2019)
D.chinensis China Freshwater Submerged decaying wood Hyde et al. (2021)
D.clematidis China; Thailand Freshwater; Terrestrial Dried stem of Clematissikkimensis; submerged decaying wood Phukhamsakda et al. (2020); Shen et al. (2021)
D.crassispora China Freshwater Submerged decaying wood Zhang et al. (2022)
D.curvularia China Freshwater Submerged decaying wood Zhang et al. (2022)
D.cylindricospora China Freshwater Submerged decaying wood Phukhamsakda et al. (2022)
D.dehongensis China; Thailand Freshwater Submerged decaying wood Hyde et al. (2019); Zhang et al. (2022)
D.dipterocarpi Thailand Terrestrial Woody litter of Dipterocarpus sp. Afshari et al. (2023)
D.effusa China Freshwater Submerged decaying wood Yang et al. (2021)
D.euseptata China Freshwater Submerged decaying wood Li et al. (2021)
D.fasciculata Thailand Freshwater Submerged decaying wood Dong et al. (2021)
D.fluminicola China Freshwater Submerged decaying wood Su et al. (2016); Luo et al. (2018)
D.fusiformis China Freshwater Submerged decaying wood Yang et al. (2021)
D.gasaensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.guanshanensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.guizhouensis China Terrestrial Decaying wood Hyde et al. (2021)
D.guttulata Thailand Freshwater Submerged decaying wood Yang et al. (2018); Luo et al. (2019)
D.hainanensis China Terrestrial Decaying wood This study
D.hyalina Thailand Freshwater Submerged decaying wood Yang et al. (2021)
D.hydei Thailand Terrestrial Decaying bamboo culms Monkai et al. (2020)
D.jinghongensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.lancangjiangensis China Freshwater Submerged decaying wood Shen et al. (2021)
D.lanceolatispora China Freshwater Submerged decaying wood This study
D.leonensis China; Malaysia Terrestrial Decaying culms of grasses or decaying branches McKenzie et al. (1995); Wu et al. (2005); Zhang et al. (2022)
D.licualae Thailand Terrestrial Decaying leaves of Licualaglabra Konta et al. (2023)
D.lignicola China; Thailand Freshwater Submerged decaying wood Luo et al. (2019); Yang et al. (2021)
D.longispora China Freshwater Submerged decaying wood Song et al. (2020)
D.longnanensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.martinii China Terrestrial Decaying branches Xia et al. (2017)
D.meilingensis China Freshwater Decaying bamboo culms Zhai et al. (2022)
D.menghaiensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.menglunensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.mengsongensis China Terrestrial Decaying branches Liu et al. (2023)
D.multiseptata Thailand Freshwater Submerged decaying wood Hyde et al. (2016); Yang et al. (2018)
D.nabanheensis China Terrestrial Decaying branches Liu et al. (2023)
D.nanchangensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.neorostrata Thailand Freshwater Submerged decaying wood Luo et al. (2019)
D.nonrostrata China Freshwater Submerged decaying wood Zhang et al. (2022)
D.obclavata Thailand Freshwater Submerged decaying wood Luo et al. (2019)
D.obpyriformis China Freshwater Submerged decaying wood Luo et al. (2018)
D.pachyconidia China Freshwater; Terrestrial Submerged decaying wood; decaying wood Ma et al. (2022); Zhang et al. (2022)
D.palmarum Thailand Terrestrial Rachis of Cocosnucifera Hyde et al. (2019)
D.phangngaensis Thailand Freshwater Submerged decaying wood Yang et al. (2018)
D.phragmiticola China Terrestrial Decaying Phragmitesaustralis Hyde et al. (2023)
D.rayongensis Thailand Freshwater Submerged decaying wood Hyde et al. (2020)
D.rostrata China Freshwater Submerged decaying wood Luo et al. (2018)
D.saprophytica Thailand Freshwater Submerged decaying wood Dong et al. (2021)
D.septata China Freshwater Submerged decaying wood Ma et al. (2022)
D.sinensis China Terrestrial Decaying branches Liu et al. (2023)
D.songkhlaensis Thailand Freshwater Submerged decaying wood Dong et al. (2021)
D.suoluoensis China Freshwater Submerged decaying wood Yang et al. (2018)
D.tectonae China; Thailand Terrestrial; Freshwater Decaying twig of Tectonagrandis; stems of dead wood; submerged decaying wood; decaying twigs of Edgeworthiachrysantha Hyde et al. (2016); Luo et al. (2018); Sun et al. (2020); Dong et al. (2021); Li et al. (2021); Zhang et al. (2022); This study
D.tectonigena Thailand Terrestrial Decaying twig of Tectonagrandis Hyde et al. (2016)
D.thailandica Thailand Terrestrial Decaying leaves of Pandanus sp. Tibpromma et al. (2018)
D.thysanolaenae China Terrestrial; Freshwater Decaying culms of Thysanolaenamaxima; Submerged decaying wood Phookamsak et al. (2019); Shen et al. (2021)
D.tropica China Terrestrial Decaying wood Ma et al. (2022)
D.verrucosa China Freshwater Submerged decaying wood Yang et al. (2021)
D.wuzhishanensis China Freshwater Submerged decaying wood Ma et al. (2022)
D.xishuangbannaensis China Terrestrial; Freshwater Decaying leaves of Pandanusutilis; submerged decaying wood Tibpromma et al. (2018); Ma et al. (2022)
D.yichunensis China Terrestrial Decaying branches of broadleaf tree Hu et al. (2023)
D.yongxiuensis China Freshwater Decaying bamboo culms Zhai et al. (2022)
D.yunjushanensis China Freshwater Decaying bamboo culms Zhai et al. (2022)
D.yunnanensis China Freshwater Submerged decaying wood Li et al. (2021)
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Supplementary Material

XML Treatment for Distoseptispora hainanensis
XML Treatment for Distoseptispora lanceolatispora
XML Treatment for Distoseptispora tectonae

Acknowledgments

The authors thank Shaun Pennycook, Manaaki Whenua – Landcare Research, New Zealand, for his guidance on the fungal nomenclature and the suggestion on naming the new taxa. The authors also thank the Guizhou Institute of Technology for its support of the experiment. Samantha Chandranath Karunarathna thanks the National Natural Science Foundation of China (Numbers 32260004) and the High-Level Talent Recruitment Plan of Yunnan Province (“High-End Foreign Experts” program) for their support.

Citation

Chen X-M, Tang X, Ma J, Liu N-G, Tibpromma S, Karunarathna SC, Xiao Y-P, Lu Y-Z (2024) Identification of two new species and a new host record of Distoseptispora (Distoseptisporaceae, Distoseptisporales, Sordariomycetes) from terrestrial and freshwater habitats in Southern China. MycoKeys 102: 83–105. https://doi.org/10.3897/mycokeys.102.115452

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 funded by the National Natural Science Foundation of China (NSFC 32360011).

Author contributions

Conceptualization - Xue-Mei Chen and Yong-Zhong Lu; data curation - Xue-Mei Chen, Xia Tang, Jian Ma, Ning-Guo Liu; formal analysis - Yuan-Pin Xiao, Xue-Mei Chen, Xia Tang, Jian Ma; funding acquisition - Yong-Zhong Lu; investigation - Saowaluck Tibpromma, Samantha C. Karunarathna, Yuan-Pin Xiao, Yong-Zhong Lu; methodology - Xue-Mei Chen, Yong-Zhong Lu; project administration - Yuan-Pin Xiao, Yong-Zhong Lu; resources - Yong-Zhong Lu, Saowaluck Tibpromma, Samantha C. Karunarathna; software - Xue-Mei Chen; supervision - Yong-Zhong Lu, Saowaluck Tibpromma, Samantha C. Karunarathna; validation - Xue-Mei Chen, Xia Tang, Jian Ma, Ning-Guo Liu; visualization - Saowaluck Tibpromma, Samantha C. Karunarathna; writing original draft - Xue-Mei Chen; writing, review and editing - Xue-Mei Chen, Xia Tang, Jian Ma, Ning-Guo Liu, Saowaluck Tibpromma, Samantha C. Karunarathna, Yuan-Pin Xiao, Yong-Zhong Lu. All authors have read and agreed to the published version of the manuscript.

Author ORCIDs

Xue-Mei Chen https://orcid.org/0009-0004-8631-0735

Xia Tang https://orcid.org/0000-0003-2705-604X

Jian Ma https://orcid.org/0009-0008-1291-640X

Ning-Guo Liu https://orcid.org/0000-0002-9169-2350

Saowaluck Tibpromma https://orcid.org/0000-0002-4706-6547

Samantha C. Karunarathna https://orcid.org/0000-0001-7080-0781

Yuan-Pin Xiao https://orcid.org/0000-0003-1730-3545

Yong-Zhong Lu https://orcid.org/0000-0002-1033-5782

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 Distoseptispora hainanensis
mycokeys.102.115452-treatment1.xml (26.8KB, xml)
XML Treatment for Distoseptispora lanceolatispora
mycokeys.102.115452-treatment2.xml (16.7KB, xml)
XML Treatment for Distoseptispora tectonae
mycokeys.102.115452-treatment3.xml (29.6KB, xml)

Data Availability Statement

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

Articles from MycoKeys are provided here courtesy of Pensoft Publishers

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