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. 2021 Nov 8;84:79–101. doi: 10.3897/mycokeys.84.71905

Two novel species and two new records of Distoseptispora from freshwater habitats in China and Thailand

Hong-Wei Shen 1,2,3, Dan-Feng Bao 1,2,4, Kevin D Hyde 2,3,5, Hong-Yan Su 1, Darbhe J Bhat 6, Zong-Long Luo 1,✉
PMCID: PMC8592981  PMID: 34790026

Abstract

During investigations into freshwater fungi from the Great Mekong Subregion, four Distoseptispora taxa were collected from China and Thailand. Based on morphological characteristics, and phylogenetic analyses of combined LSU, ITS, SSU, TEF1-α, and RPB2 sequence data, two new species Distoseptisporabangkokensis and D.lancangjiangensis are introduced, and two known species D.clematidis and D.thysanolaenae were first reported in freshwater habitat. Illustrations and descriptions of these taxa are provided, along with comparisons with extant taxa in the genus.

Keywords: 2 new taxa, Distoseptisporales, freshwater fungi, morphology, phylogeny, taxonomy

Introduction

Distoseptisporaceae was introduced by Su et al. (2016) based on morphological and phylogenetic analyses, with Distoseptispora as type genus. Distoseptisporaceae is placed in Distoseptisporales, which was introduced by Luo et al. (2019), and currently comprises two families, Aquapteridosporaceae and Distoseptisporaceae (Luo et al. 2019; Wijayawardene et al. 2020; Hyde et al. 2021). Species of both families are commonly reported from freshwater habitats (Yang et al. 2015, 2018; Su et al. 2016; Li et al. 2021; Hyde et al. 2016a, 2019, 2020; Luo et al. 2018, 2019; Song et al. 2020; Dong et al. 2021).

Distoseptispora as a single genus in Distoseptisporaceae was introduced by Su et al. (2016) with D.fluminicola as the type species. The genus is characterized by monoblastic, cylindrical, conidiogenous cells, with percurrent proliferation, acrogenous, solitary, brown or yellowish/reddish brown, olivaceous, distoseptate or euseptate, cylindrical, obclavate, rostrate conidia, truncate base, with rounded apices, basal cell with a cross wall and basal scar. This genus is not known for its sexual morph (Su et al. 2016; Yang et al. 2018; Hyde et al. 2019, 2020; Luo et al. 2019; Sun et al. 2020). Currently, 32 species are accepted in the genus of which 13 from terrestrial habitats and 19 were reported from freshwater environments (Su et al. 2016; Hyde et al. 2016a, 2019, 2020; Xia et al. 2017; Yang et al. 2018; Luo et al. 2018, 2019; Monkai et al. 2020; Song et al. 2020; Sun et al. 2020; Li et al. 2021; Index Fungorum 2021http://www.indexfungorum.org).

During our ongoing study of freshwater fungi along the north-south gradient in the Asian/Australian region (Hyde et al. 2016b), we collected four species in the genus. Two new species, Distoseptisporabangkokensis and D.lancangjiangensis, are introduced in this study, D.clematidis and D.thysanolaenae are newly recorded from freshwater habitats for the first time in China. Morphological descriptions and illustrations of the species and an updated multi-gene phylogenetic tree are provided to reveal their taxonomic position among the species in the Distoseptisporales, and also provided the comparison of morphological characteristics, habitats and hosts information of species newly added to Distoseptispora after Monkai et al. (2020) (Table 2).

Table 2.

Comparison of morphological characteristic, habitats and hosts’ information of species added to Distoseptispora after Monkai et al. (2020) (for other species see Monkai et al. 2020).

Species Conidiophore (μm) Conidia (μm) Conidia septation Conidia characteristic Habitat Host Reference
Distoseptisporabangkokensis 37–55 × 3–4 400–568 × 13–16 Multi-distoseptate Elongate, obclavate, rostrate, dark olivaceous to dark brown Freshwater Unidentified submerged wood This study
D.lancangjiangensis 30–41 × 5–6 83–220 × 12–14 16–41-distoseptate Obclavate, cylindrical, elongated, straight or curved, brown to greenish-brown Freshwater Unidentified submerged wood This study
D.euseptata 19–28 × 4–5 37–54 × 8–9 4–7-euseptate Obpyriform to obclavate, straight or curved, olivaceous Freshwater Unidentified submerged wood Li et al. 2021
D.fasciculata 12–16 × 5–6 46–200 × 10–16.5 10–40-distoseptate Subcylindrical to obclavate, mostly curved, olivaceous when young, dark brown when mature Freshwater Unidentified submerged wood Dong et al. 2021
D.longispora 17–37 × 6–10 189–297 × 16–23 31–56-distoseptate Obclavate, elongated, straight or slightly curved, to yellowish brown Freshwater Unidentified submerged wood Song et al. 2020
D.saprophytica 50–140 × 3.2–4.2 14.5–30 × 4.5–7.5 2–6-distoseptate Subcylindrical to obclavate, straight or curved, olivaceous to brown Freshwater Unidentified submerged wood Dong et al. 2021
D.songkhlaensis 70–90 × 4–5.5 44–125 × 9–14.5 9–16-distoseptate Obclavate, straight or curved, olivaceous to brown Freshwater Unidentified submerged wood Dong et al. 2021
D.yunnanensis 131–175 × 6–7 58–108 × 8–10 6–10-euseptate Obclavate, rostrate, straight or slightly curved, mid olivaceous to brown Freshwater Unidentified submerged wood Li et al. 2021
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Materials and methods

Isolation and morphology

Specimens of submerged decaying wood were collected from Dulongjiang, Nanpanjiang, Lancangjiang and Chao Phraya River in China and Thailand respectively. Multiple samples will be collected at each collection site at different times, allowing more strains to be obtained for each species. Methods of morphological observation and isolation follow Luo et al. (2018) and Senanayake et al. (2020). IFW (Tarosoft(R) Image Frame Work) was used for measurement of photomicrograph, and Adobe Photoshop CS5 software was used to process images for making photo-plates (Adobe Systems Inc., USA). Single spore isolation was performed according to the following steps: The conidia suspension from specimens, absorbed with a sterilized pipette, was placed on potato dextrose agar (PDA) and incubated at room temperature overnight. Germinated conidia were transferred to new PDA/MEA (Beijing land bridge technology CO., LTD., China) plates and incubated in an incubator at room temperature (25 °C). Specimens were deposited in the Kunming Institute of Botany, Academia Sinica herbarium (KUN-HKAS), and Mae Fah Luang University herbarium (MFLU). Cultures were deposited in the Dali University Culture Collection (DLUCC), China General Microbiological Culture Collection Center (CGMCC), and Mae Fah Luang University Culture Collection (MFLUCC). Facesoffungi number was obtained as described in Jayasiri et al. (2015) and Index Fungorum number was also registered (http://www.indexfungorum.org/Names/Names.asp). In this study, multiple samples were collected for each sample site and related environment, but unfortunately, there were still no more strains for the two new species in the paper.

DNA extraction, PCR amplification, and sequencing

DNA extraction, PCR amplification, sequencing and phylogenetic analysis follow Dissayanake et al. (2020) with the following modifications. Fungal mycelia (200–500 mg) were scraped from grown on PDA/MEA plates using sterile scalpel, transferred to microcentrifuge tube with sterilized needles, and then grind with liquid nitrogen or quartz sand to break the cells. DNA was extracted using the TreliefTM Plant Genomic DNA Kit (TSP101) according to the manufacturer’s instructions.

Five gene regions, LSU, ITS, SSU, TEF1-α, and RPB2 were amplified using LR0R/LR5, ITS5/ITS4, NS1/NS4, 983F/EF1-2218R, and RPB2-5F/RPB2-7cR (Vilgalys and Hester 1990; White et al. 1990; Liu et al. 1999) primer pairs respectively. Primer sequences are available at the WASABI database at the AFTOL website (aftol.org). The PCR mixture contained 12.5 μL of 2 × Power Taq PCR Master Mix (a premix and ready to use solution, including 0.1 Units/μL Taq DNA Polymerase, 500μm dNTP Mixture each (dATP, dCTP, dGTP, dTTP), 20 mm Tris-HCl pH 8.3, 100 Mm KCl, 3 mM MgCl2, stabilizer and enhancer), 1 μL of each primer including forwarding primer and reverse primer (10 μm), 1 μL template DNA extract and 9.5 μL deionized water (Luo et al. 2018). The PCR cycling conditions of LSU, ITS, SSU and TEF1-α were as follows: 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 30s, annealing at 55 °C for 50s, elongation at 72 °C for 1 min, and a final extension at 72 °C for 10 min. The PCR thermal cycle of RPB2 has a total of 40 cycles, and the conditions are as follows: initially denature at 95 °C for 5 min, and then enter 40 cycles: denaturation at 95 °C for 1 min, annealing at 52 °C for 2 min, extension at 72 °C for 90s, and finally at 72 °C for 10 min. PCR products were then purified using minicolumns, purification resin, and buffer according to the manufacturer’s protocols (Amersham product code: 27–9602–01). The sequences were carried out at Beijing Tsingke Biotechnology Co., Ltd. (Beijing, P.R. China).

Phylogenetic analysis

Preliminary identification of genes obtained from fresh strains by GenBank database. The LSU, ITS, SSU, TEF1-α, and RPB2 used for phylogenetic analysis are selected based on the preliminary identification results and the related publications (Yang et al. 2018; Monkai et al. 2020). The sequences were aligned using MAFFT online service: Multiple alignment program for amino acid or nucleotide sequences MAFFT version 7 (Katoh and Standley 2013: http://mafft.cbrc.jp/alignment/server/index.html), and edited manually in BioEdit v. 7.0 (Hall 1999). The sequence dataset was combined using SquenceMatrix v.1.7.8 (Vaidya et al. 2011). The alignment formats were change to PHYLIP and NEXUS formats by ALigment Transformation EnviRonment (ALTER) website (http://sing.ei.uvigo.es/ALTER/).

Maximum likelihood (ML) analysis was carried out using the RAxML-HPC2 on XSEDE (8.2.12) (Stamatakis 2006; Stamatakis et al. 2008) of CIPRES Science Gateway website (Miller et al. 2010: http://www.phylo.org/portal2) and the estimated proportion of invariant sites is (GTRGAMMA+I) model.

Bayesian analyses were performed in MrBayes 3.2.6 (Ronquist et al. 2012) and the best-fit model (LSU, ITS, SSU, TEF1-α, and RPB2 are all GTR+I+G) of sequences evolution was estimated via MrModeltest 2.2 (Guindon and Gascuel 2003; Nylander 2004; Darriba et al. 2012). The Markov Chain Monte Carlo (MCMC) sampling approach was used to calculate posterior probabilities (PP) (Rannala and Yang 1996). Bayesian analyses of six simultaneous Markov chains were run for 10000000 generations with trees sampled every 1000 generations.

Phylogenetic trees were visualized using FigTree v1.4.0 (Rambaut 2012: http://tree.bio.ed.ac.uk/software/figtree/), editing and typesetting using Adobe Illustrator (AI) (Adobe Systems Inc., the United States). The new sequences were submitted in GenBank and the strain information used in this paper is provided in Table 1. The alignments and phylogenetic trees were deposited in TreeBASE (http://www.treebase.org/, accession number: 28758).

Table 1.

Strains used for phylogenetic analysis and their corresponding GenBank numbers. The type strain are in bold font.

Species Source GenBank accession number Reference
LSU ITS TEF1-α RPB2 SSU
Aquapteridosporafusiformis MFLUCC 18-1606 MK849798 MK828652 MN194056 – – Luo et al. (2019)
A.lignicola MFLUCC 15-0377 KU221018 – – – – Yang et al. (2015)
Distoseptisporaadscendens HKUCC 10820 DQ408561 – – DQ435092 – Shenoy et al. (2006)
D.appendiculata MFLUCC 18-0259 MN163023 MN163009 MN174866 – – Luo et al.(2019)
D.aquatica MFLUCC 15-0374 KU376268 MF077552 – – – Su et al. (2016)
D.bambusae MFLUCC 20-0091 MT232718 MT232713 MT232880 MT232881 MT232716 Sun et al. (2020)
D.bambusae MFLUCC 14-0583 MT232717 MT232712 – MT232882 – Sun et al. (2020)
D.bangkokensis MFLUCC 18-0262 MZ518206 MZ518205 – – MZ518208 This study
D.cangshanensis MFLUCC 16-0970 MG979761 MG979754 MG988419 – – Luo et al. (2018)
D.caricis CBS 146041 MN567632 MN562124 – MN556805 – Crous et al. (2019)
D.clematidis MFLUCC 17-2145 MT214617 MT310661 – MT394721 MT226728 Phukhamsakda et al. (2020)
D.clematidis KUN-HKAS 112708 MW879523 MW723056 MW729784 – MW774580 This study
D.dehongensis KUMCC 18-0090 MK079662 MK085061 MK087659 – – Hyde et al. (2019)
D.euseptata MFUCC 20-0154 MW081544 MW081539 – MW151860 – Li et al. (2021)
D.euseptata DLUCC S2024 MW081545 MW081540 MW084994 MW084996 – Li et al. (2021)
D.fasciculata KUMCC 19-0081 MW287775 MW286501 MW396656 – – Dong et al. (2021)
D.fluminicola MFLUCC 15-0417 KU376270 MF077553 – – – Su et al. (2016)
D.guttulata MFLUCC 16-0183 MF077554 MF077543 MF135651 – MF077532 Yang et al. (2018)
D.hydei MFLUCC 20-0115 MT742830 MT734661 – MT767128 – Monkai et al. (2020)
D.lancangjiangensis KUN-HKAS 112712 MW879522 MW723055 – MW882260 – This study
D.leonensis HKUCC 10822 DQ408566 – – DQ435089 – Shenoy et al. (2006)
D.lignicola MFLUCC 18-0198 MK849797 MK828651 – – MK828318 Luo et al. (2019)
D.longispora HFJAU 0705 MH555357 MH555359 – – MH555431 Song et al. (2020)
D.martinii CGMCC 3.18651 KX033566 KU999975 – – KX033537 Xia et al. (2017)
D.multiseptata MFLUCC 16-1044 MF077555 MF077544 MF135652 MF135644 MF077533 Yang et al. (2018)
D.multiseptata MFLUCC 15-0609 KX710140 KX710145 MF135659 – NG_065693 Hyde et al. (2016)
D.neorostrata MFLUCC 18-0376 MN163017 MN163008 – – – Luo et al. (2019)
D.obclavata MFLUCC 18-0329 MN163010 MN163012 – – – Luo et al. (2019)
D.obpyriformis MFLUCC 17-01694 MG979764 – MG988422 MG988415 – Luo et al. (2018)
D.obpyriformis DLUCC 0867 MG979765 MG979757 MG988423 MG988416 – Luo et al. (2018)
D.palmarum MFLUCC 18-1446 MK079663 MK085062 MK087660 MK087670 MK079661 Hyde et al. (2019)
D.phangngaensis MFLUCC 16-0857 MF077556 MF077545 MF135653 – MF077534 Yang et al. (2018)
D.rayongensis MFLUCC 18-0415 MH457137 MH457172 MH463253 MH463255 MH457169 Hyde et al. (2012)
D.rostrata MFLUCC 16-0969 MG979766 MG979758 MG988424 MG988417 – Luo et al. (2018)
D.saprophytica MFLUCC 18-1238 MW287780 MW286506 MW396651 MW504069 – Dong et al. (2021)
D.songkhlaensis MFLUCC 18-1234 MW287755 MW286482 MW396642 – – Dong et al. (2021)
D.suoluoensis MFLUCC 17-0224 MF077557 MF077546 MF135654 – MF077535 Yang et al. (2018)
D.suoluoensis MFLUCC 17-1305 MF077558 MF077547 – – MF077536 Yang et al. (2018)
D.tectonae MFLUCC 12-0291 KX751713 KX751711 KX751710 KX751708 – Hyde et al. (2016)
D.tectonae*1 MFLUCC 16-0946 MG979768 MG979760 MG988426 MG988418 – Luo et al. (2018)
D.tectonigena MFLUCC 12-0292 KX751714 KX751712 – KX751709 – Hyde et al. (2016)
D.thailandica MFLUCC 16-0270 MH260292 MH275060 MH412767 – MH260334 Tibpromma et al. (2018)
D.thysanolaenae KUN-HKAS 102247 MK064091 MK045851 MK086031 – – Phookamsak et al. (2019)
D.thysanolaenae KUN-HKAS 112710 MW879524 MW723057 MW729783 – – This study
D.xishuangbannaensis KUMCC 17-0290 MH260293 MH275061 MH412768 MH412754 MH260335 Tibpromma et al. (2018)
D.yunnanensis MFLUCC 20-0153 MW081546 MW081541 MW084995 MW151861 – Li et al. (2021)
Myrmecridiumaquaticum MFLUCC 15-0366 MK849804 – – – MK828323 Luo et al. (2019)
M.aquaticum S-1158 MK849803 MK828656 MN194061 MN124540 MK828322 Luo et al. (2019)
M.banksiae CBS 132536 JX069855 JX069871 – – – Crous et al. (2012)
Pseudostanjehughesiaaquitropica MFLUCC 16-0569 MF077559 MF077548 MF135655 – MF077537 Yang et al. (2018)
P.lignicola MFLUCC 15-0352 MK849787 MK828643 MN194047 MN124534 – Luo et al. (2019)
Sporidesmiumdulongense MFLUCC 17-0116 MH795817 MH795812 MH801191 MH801190 – Luo et al. (2019)
S.lageniforme DLUCC 0880 MK849782 MK828640 MN194044 MN124533 – Luo et al. (2019)
S.pyriformatum MFLUCC 15-0620 KX710141 KX710146 MF135662 MF135649 – Hyde et al. (2016)
S.thailandense MFLUCC 15-0617 MF077561 MF077550 MF135657 – – Yang et al. (2018)
S.thailandense MFLUCC 15-0964 MF374370 MF374361 MF370957 MF370955 – Zhang et al. (2017)
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*1 Ex-type strain of Distoseptisporasubmersa.

Results

Phylogenetic analysis

The dataset composed of LSU (1–744 bp), ITS (745–1310 bp), TEF1-α (1311–2161 bp), RPB2 (2162–3178 bp), and SSU (3179–4199 bp) gene, comprising a total of 4199 characters (including gaps), including 56 taxa with Pseudostanjehughesiaaquitropica (MFLUCC 16-0569) and P.lignicola (MFLUCC 15-0352) as the outgroup taxa (Figure 1). The ML and BI phylogenetic analyses produced similar topology. The combined dataset analysis of RAxML generates a best-scoring tree (Figure 1), with the final ML optimization likelihood value of -30393.557997. The aligned matrix had 1624 distinct alignment patterns, with 36.44% completely undetermined characters or gaps. The base frequency and rate are as follows: A = 0.243915, C = 0.259360, G = 0.279029, T = 0.217696; rate AC = 1.166355, AG = 2.813539, AT = 1.110401, CG = 0.796371, CT = 5.621229, GT = 1.000000; gamma distribution shape: α = 0.221933. Bootstrap support values with a maximum likelihood (ML) greater than 70%, and Bayesian posterior probabilities (PP) greater than 0.97 are given above the nodes.

Figure 1.

Figure 1.

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Maximum likelihood (ML) tree is based on combined of LSU, ITS, SSU, TEF1-α, and RPB2 sequence data. Bootstrap support values with an ML greater than 70% and Bayesian posterior probabilities (PP) greater than 0.97 given above the nodes, shown as “ML/PP”. The tree is rooted with Pseudostanjehughesiaaquitropica (MFLUCC 16-0569) and P.lignicola (MFLUCC 15-0352). New species are indicated in red and type strains are in bold.

The phylogenetic tree shows that the new species Distoseptisporabangkokensis (MFLUCC 18-0262) was placed as a sister taxon to D.bambusae (MFLUCC 14-0583 and MFLUCC 20-0091), D.dehongensis (KUMCC 18-0090), D.euseptata (MFUCC 20-0154 and DLUCC S2024), D.lancangjiangensis (KUN-HKAS 112712), D.suoluoensis (MFLUCC 17-0224 and MFLUCC 17-1305), D.thysanolaenae (KUN-HKAS 102247 and KUN-HKAS 112710), and D.yunnanensis (MFLUCC 20-0153) with low bootstrap support with low bootstrap support (Figure 1), whereas D.lancangjiangensis clustered with D.suoluoensis with 97%ML/0.98PP support. Distoseptisporathysanolaenae (KUN-HKAS 112710) and D.clematidis (KUN-HKAS 112708) clustered with the ex-type strain of D.thysanolaenae (KUN-HKAS 102247) and D.clematidis (MFLUCC 17-2145), respectively, with 100%ML/1.00PP and 97%ML/0.99PP bootstrap support.

Taxonomy

Distoseptispora bangkokensis

H.W. Shen, D.F. Bao, K.D. Hyde & Z.L. Luo sp. nov.

E08487C9-F52C-559D-B571-BFE767D78DD4

Index Fungorum Number No: IF558556

Facesoffungi Number No: FoF09993

Figure 2

Figure 2.

Figure 2.

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Distoseptisporabangkokensis (MFLU 21-0110, holotype) A colonies on the substratum B conidiophores C conidiophores with conidia D conidiogenous cell E-G conidia H germinating conidium Scale bars: 20 μm (B, C, H); 10 μm (D); 50 μm (E-G).

Etymology.

Referring to the collecting location, Bangkok, Thailand.

Holotype.

MFLU 21-0110

Description.

Saprobic on submerged wood in freshwater stream. Sexual morph: Undetermined. Asexual morph: Colonies effuse, glistening, hairy, brown to dark brown. Mycelium partly superficial in the substratum, composed of hyaline to pale brown, septate, branched hyphae. Conidiophores 37–55 × 3–4 μm (x¯ = 46 × 3 μm, n = 15) macronematous, mononematous, solitary or in a small group of 2–4, cylindrical, straight or slightly flexuous, 3–8-septate, dark brown, paler at the apical part, rounded at the apex. Conidiogenous cells 6–8 × 3–4 μm (x¯ = 7 × 3 μm, n = 15), integrated, terminal, monoblastic, cylindrical, brown. Conidia 400–568 × 13–16 μm (x¯ = 484 × 15 μm, n = 20), 6–7 μm at the narrowest apical region, acrogenous, solitary, elongate, obclavate, rostrate, multi-distoseptate, tapering towards the apex, truncate at the base, rounded at apex, dark olivaceous to dark brown, straight or slightly curved, guttulate, thick-walled, smooth, conidia percurrent proliferation which forms another conidium at the apex.

Culture characteristics. Conidia cultivated on PDA within 12h and germ tubes produced at the ends. Colonies on PDA, reaching 6 cm in 1 month at room temperature (25 °C). Mycelium loose, flocculent, smooth edge, brown to dark brown, dark brown on the reverse.

Material examined.

Thailand, Bangkok Province, Khwaeng Phra Khanong Nuea, 13°42'41"N; 100°36'03"E, on submerged decaying wood, 1 October 2017, Zonglong Luo, S–3083 (MFLU 21-0110, holotype), ex-type living culture (MFLUCC 18-0262).

Notes.

Distoseptisporabangkokensis is comparable to D.cangshanensis and D.multiseptata in having elongate, obclavate, or rostrate conidia (Su et al. 2016; Hyde et al. 2016a; Yang et al. 2018). However, D.bangkokensis has shorter and narrower conidiophores than those of D.cangshanensis (37–55 × 3–4 μm vs. 44–68 × 4–8 μm), but has longer conidia (400–568 μm vs. 58–166 μm); D.multiseptata (MFLU 17-0856) is similar to D.bangkokensis in conidial morphology, with conidia mostly 300–600 μm long (up to 700 μm) and significantly longer than those of the holotype (up to 380 μm long). However, Yang et al. (2018) did not give a detailed description of D.multiseptata (MFLU 17-0856). Phylogenetic analyses showed that D.bangkokensis clustered with D.bambusae, D.dehongensis, D.euseptata, D.lancangjiangensis, D.suoluoensis, D.thysanolaenae, and D.yunnanensis with low bootstrap support (26%ML/0.53PP, Figure 1). Distoseptisporabangkokensis is distoseptate conidia, and it is easily distinguished from D.bambusae, D.euseptata, D.lancangjiangensis, D.suoluoensis, and D.yunnanensis, which are euseptate. Distoseptisporabangkokensis is resemble to D.dehongensis and D.thysanolaenae in having obclavate, distoseptae conidia, but are distinguished by conidia characteristics, D.bangkokensis has elongate, obclavate, rostrate, multi-distoseptat, and longer conidia than D.dehongensis (400–568 × 13–16 μm vs. 17–30 × 7.5–10 μm) and D.thysanolaenae (400–568 × 13–16 μm vs. 30–70 × 5–8 μm), respectively.

Distoseptispora lancangjiangensis

H.W. Shen, H.Y. Su, K.D. Hyde & Z.L. Luo sp. nov.

BB47E43B-AC2E-525D-A336-5ED94C89D927

Index Fungorum Number No: IF558555

Facesoffungi Number No: FoF09994

Figure 3

Figure 3.

Figure 3.

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Distoseptisporalancangjiangensis (KUN-HKAS 112712, holotype) A colonies on the substratum B conidiophore and conidium C-E conidiophores F, G conidiogenous cells H conidiogenous cell with conidium I-Q conidia R germinating conidium S, T culture on PDA. Scale bars: 50 μm (B-E); 20 μm (F-R).

Etymology.

Referring to the collecting location, Lancangjiang River in China.

Holotype.

KUN-HKAS 112712

Description.

Saprobic on submerged wood in freshwater River. Sexual morph: Undetermined. Asexual morph: Colonies effuse, hairy, glistening, brown to dark. Mycelium partly immersed in the substratum, composed of hyaline to pale brown, septate, branched hyphae. Conidiophores 144–204 × 5–6 μm (x¯ = 175 × 6 μm, n = 20) macronematous, mononematous, solitary, inflate at the base, cylindrical, straight or slightly flexuous, 6–11-septate, dark brown, hyaline and rounded at apex. Conidiogenous cells 12–24 × 4–5 μm (x¯ = 18 × 5 μm, n = 20) integrated, terminal, monoblastic, cylindrical, brown. Conidia 64–84 × 9–10 μm (x¯ = 74 × 10 μm, n = 20), acrogenous, solitary, narrowly obclavate or obspathulate, tracted at base, tapering towards apex, 3–10-euseptate, brown to dark brown, thin-walled, becoming paler or hyaline towards apex, guttulate, with a darkened scar at base, smooth-walled.

Culture characteristics.

Conidia cultivated on PDA within 12h and germ tubes produced at the apex. Colonies on PDA, reaching 4.5 cm in 1 month at room temperature (25 °C). Mycelium loose, flocculent, smooth edges, convex middle, pale brown to dark brown on the surface of PDA. Smooth, black on the reverse.

Material examined.

China, Yunnan Province, Dali City, Lancangjiang River, 22°36'36"N; 100°37'59"E, on submerged decaying wood, 20 July 2017, Qishan Zhou and Qingxiong Ruan S–1864 (KUN-HKAS 112712, holotype; MFLU 21-0111, isotype), ex-type living culture (DLUCC 1864 = CGMCC 3.20265).

Notes.

Phylogenetic analysis showed that Distoseptisporalancangjiangensis clustered as a sister taxon to D.suoluoensis with 97%ML/0.98PP support. Distoseptisporalancangjiangensis is similar to D.suoluoensis in having long conidiophores, monoblastic conidiogenous cells, and obclavate to rostrate, euseptate conidia. However, D.suoluoensis has yellowish-brown or dark olivaceous, verrucose conidia, while in D.lancangjiangensis conidia are brown to dark brown and smooth-walled. Moreover, D.lancangjiangensis has smaller conidia than those of D.suoluoensis (64–84 × 9–10 μm vs. 80–125 × 8–13 μm) (Yang et al. 2018). Distoseptisporalancangjiangensis and D.bambusae have similar conidial shapes, but D.lancangjiangensis having longer conidia (64–84 × 9–10 μm vs. 45–74 × 5.5–10 μm) and longer conidiophores (144–204 × 5–6 μm vs. 40–96 × 4–5.5 μm). Furthermore, D.bambusae has polyblastic or monoblastic conidiogenous cells and olivaceous or brown conidia, while D.lancangjiangensis only has monoblastic conidiogenous cells and brown to dark brown conidia (Sun et al. 2020).

Distoseptispora clematidis

Phukhams., M.V. de Bult & K.D. Hyde, in Phukhamsakda et al., Fungal Diversity 102: 168 (2020)

AC7F9A70-BD17-5567-BC09-C62CA24F0F50

Index Fungorum Number No: IF557301

Facesofungi Number No: FoF07261

Figure 4

Figure 4.

Figure 4.

Open in a new tab

Distoseptisporaclematidis (KUN-HKAS 112708) A colonies on the substratum B-C conidiophores with conidia D conidiogenous cells E-H conidia I germinating conidium J culture on PDA Scale bars: 30 μm (B, C, E-I); 20 μm (D).

Description.

Saprobic on submerged wood in freshwater River. Sexual morph: Undetermined. Asexual morph: Colonies on the substratum superficial, effuse, scattered, hairy, dark brown. Mycelium partly immersed in substrate, composed of branched, smooth, septate, brown to dark brown hyphae. Conidiophores 30–41 × 5–6 μm (x¯ = 36 × 6 μm, n = 15), macronematous, mononematous, single or in a small group, straight or slightly flexuous, unbranched, septate, erect, 2–4-septate, cylindrical, smooth, dark brown to brown. Conidiogenous cells 7–9 × 5–6 μm (x¯ = 8 × 5 μm, n = 15), monoblastic, integrated, determinate, terminal, cylindrical, pale brown to brown. Conidia 83–220 × 12–14 μm (x¯ = 151 × 13 μm, n = 20), acrogenous, solitary, obclavate, cylindrical, elongated, straight or curved, truncate at base, rounded at apex, 16–41-distoseptate, slightly constricted at some septa, smooth, brown to greenish-brown, thick-walled.

Culture characters.

Conidia cultivated on PDA within 12h and germ tubes produced at the ends. Colonies on PDA, attaining 4 cm after 1 month at room temperature (25 °C), gray at first, later becoming dark gray, loose, flocculent, smooth edge, dark brown on the reverse.

Material examined.

China, Yunnan Province, Kunming City, Yiliang County, Nanpanjiang River, 24°38'28"N; 103°09'38"E, on submerged decaying wood, 12 June 2018; Hongwei Shen and Xiu He, S–1797 (KUN-HKAS 112708), living culture (DLUCC 1797).

Notes.

Our new isolate clustered with the ex-type strain of Distoseptisporaclematidis (MFLU 17-1501) (Phukhamsakda et al. 2020) with 97%ML/0.99PP bootstrap support (Figure 1). Distoseptisporaclematidis (MFLU 17-1501) was collected on dead culms of Thysanolaenamaxima (Roxb. ex Hornem.) Honda in Yunnan Province, China. Based on morphological analysis, the size and shape of the conidia and conidiophores of our new isolate are similar to D.clematidis. Therefore, we identified our new isolate as D.clematidis and it is a new record from freshwater habitats in China.

Distoseptispora thysanolaenae

Goonas., Dayarathne, Phookamsak & K.D.Hyde, in Phookamsak et al., Fungal Diversity 95: 126 (2019)

68BD6629-9E92-5AC8-BE36-175569F80A7A

Index Fungorum Number No: IF555408

Facesoffungi Number No: FoF05011

Figure 5

Figure 5.

Figure 5.

Open in a new tab

Distoseptisporathysanolaenae (KUN-HKAS 112710) A colonies on the substratum B-D conidiophores with conidia E, F conidiogenous cells G-N conidia O germinating conidium P, Q culture on PDA Scale bars: 30 μm (B-D); 10 μm (E, F); 20 μm (G-O).

Description.

Saprobic on submerged wood in freshwater River. Sexual morph: Undetermined. Asexual morph: Colonies on the substratum superficial, effuse, scattered, hairy, dark brown. Mycelium partly immersed, composed of branched, septate, smooth, brown to dark brown hyphae. Conidiophores 41–59 × 4–5 μm (x¯ = 50 × 5 μm, n = 20) macronematous, mononematous, unbranched, single, erect, straight or slightly curved, smooth, 3–6-septate, pale brown to brown. Conidiogenous cells monoblastic, integrated, determinate, terminal, dark brown, cylindrical. Conidia 46–87 × 9–12 μm (x¯ = 67 × 10 μm, n = 25) acrogenous, solitary, dry, smooth, obclavate, elongated, straight or slightly curved, truncate at base, tapering towards apex, 6–19-septate, dark grayish-brown to light yellow-green, thick-walled.

Culture characteristics.

Conidia cultivated on PDA within 12 h and germ tubes produced at the apex. Colonies on PDA, reaching 6 cm after 6 weeks at room temperature (25 ℃). Mycelium loose, flocculent, neat edges, convex in middle, pale brown to dark brown. Black, smooth on the back.

Material examined.

China, Yunnan Province, Lushui City, Nujiang River, 26°23'12"N; 98°53'94"E, on submerged decaying wood, 3 May 2016, Zonglong Luo and Songming Tang, S-876 (KUN-HKAS 112710), living culture (DLUCC 876 = KUNCC 21-10710)

Notes.

Our new collection is identical to Distoseptisporathysanolaenae in characters of the conidiophores, conidiogenous cell, and conidia (Phookamsak et al. 2019). Furthermore, our new isolate phylogenetically clusters with the ex-type strain of D.thysanolaenae (KUN-HKAS 102247) with 100%ML/1.00PP support (Figure 1). Distoseptisporathysanolaenae was collected from terrestrial habitats in China, while, our new isolate was collected from freshwater habitat in China. Therefore, we identified our new collection as D.thysanolaenae, and it is new to freshwater habitats in China.

Discussion

Distoseptispora has been reported from both freshwater and terrestrial habitats. Of these, species have been collected from freshwater environments (Su et al. 2016; Hyde et al. 2016a, 2019, 2020; Luo et al. 2018; Xia et al. 2017, 2019; Yang et al. 2018; Tibpromma et al. 2018; Crous et al. 2019; Phookamsak et al. 2019; Monkai et al. 2020; Phukhamsakda et al. 2020; Song et al. 2020; Sun et al. 2020; Li et al. 2021). To date, 18 species of Distoseptispora have been reported from Thailand, 14 species from China. In this study, we collected four distoseptispora-like taxa from rivers and streams in China and Thailand. Phylogenetic analysis showed that all four species were well-placed in Distoseptispora (Figure 1). Two new species and records are introduced based on morphological and phylogenetic analysis.

Species of Distoseptispora are highly diverse in morphology, especially the conidial shape. Conidia of most species are obclavate to cylindrical or rostrate (e.g. D.aquatica, D.tectonae, and D.suoluoensis), but a few are ellipsoid to subglobose (e.g. D.martinii), lanceolate (e.g. D.guttulata and D.multiseptata), and some species have conidia with a sheath at the apex (e.g. D.appendiculata) (Hyde et al. 2016a; Su et al. 2016; Xia et al. 2017; Yang et al. 2018; Luo et al. 2018, 2019). Some species also differ in the conidiogenous cells (D.palmarum, D.dehongensis, and D.bambusae are monoblastic or polyblastic, while the others are monoblastic) and conidial septate (D.bambusae, D.euseptatensis, D.guttulata, D.lignicola, D.rayongensis, D.suoluoensis, and D.yunnanensis are euseptate, while other species are distoseptate) (Yang et al. 2018; Hyde et al. 2019; Luo et al. 2019; Sun et al. 2020; Dong et al. 2021; Li et al. 2021).

Based on the key morphological characteristics, viz. conidiophores, conidiogenous cells, and conidia, Subramanian (1992) redisposed seven genera, viz., Sporidesmium, Polydesmus, Sporidesmiella, Stanjehughesia, Repetophragma, Penzigomyces, and Ellisembia to accommodate several Sporidesmium-like taxa. Based on multi-gene phylogenetic analysis and morphology, Su et al. (2016) introduced a new Sporidesmium-like genus Distoseptispora. Some Sporidesmium-like taxa were introduced in different lineages and synonymized Ellisembia under Sporidesmium. Although Distoseptispora was only introduced from submerged wood in freshwater habitat in 2016 (Su et al. 2016), the genus has previously been reported from both freshwater and terrestrial habitats as species in other genera. For example, Cai et al. (2002), Ho et al. (2001, 2002) and Luo et al. (2004) reported Distoseptispora as other species (Ellisembia, Sporidesmiella, and Sporidesmium) from submerged wood in freshwater habitats, and Kodsueb et al. (2016), Mena-Portales et al. (2016) and Zhou et al (2001) reported from terrestrial habitats. However, none of these records had molecular data and it is impossible to consider the placement of these isolates. In these species distoseptispora/sporidesmium-like genera, it is therefore better to describe taxa based on molecular data.

Based on phylogenetic analysis, Xia et al. (2017) transferred Acrodictysmartinii to Distoseptispora as Distoseptisporamartinii. The species is characterized by solitary erect, unbranched conidiophores, monoblastic conidiogenous cells with percurrent extensions and subhyaline to pale brown and solitary, transversal ellipsoid, oblate or subglobose, muriform conidia, separated by septa, sometimes with pores in the septa and pale brown to brown. However, the current understanding of Distoseptisporaceae, D.martinii is significantly different from other Distoseptispora taxa; thus, needs to be verified in the future (Luo et al. 2018; Sun et al. 2020).

Supplementary Material

XML Treatment for Distoseptispora bangkokensis
XML Treatment for Distoseptispora lancangjiangensis
XML Treatment for Distoseptispora clematidis
XML Treatment for Distoseptispora thysanolaenae

Acknowledgment

We would like to thank the National Natural Science Foundation of China (Project ID: 32060005 and 31970021), and Fungal Diversity Conservation and Utilization Innovation Team of Dali University (ZKLX2019213) for financial support. Kevin D Hyde thanks the Thailand Research Fund for the grant (RDG6130001MS), Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion. Hongwei Shen thanks Saranyaphat Boonmee for her help in sample collection and herbarium deposit. Qishan Zhou, Qingxiong Ruan, Songming Tang, and Xiu He are thanked for their help on sample collection. We are grateful to Yanmei Zhang, Longli Li, and Wenli Li for their help on DNA extraction and PCR amplification.

Citation

Shen H-W, Bao D-F, Hyde KD, Su H-Y, Bhat DJ, Luo Z-L (2021) Two novel species and two new records of Distoseptispora from freshwater habitats in China and Thailand. MycoKeys 84: 79–101. https://doi.org/10.3897/mycokeys.84.71905

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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 bangkokensis
mycokeys.84.71905-treatment1.xml (31.8KB, xml)
XML Treatment for Distoseptispora lancangjiangensis
mycokeys.84.71905-treatment2.xml (14.8KB, xml)
XML Treatment for Distoseptispora clematidis
mycokeys.84.71905-treatment3.xml (13.4KB, xml)
XML Treatment for Distoseptispora thysanolaenae
mycokeys.84.71905-treatment4.xml (20.2KB, xml)

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