Three New Species, Two New Records and Four New Collections of Tubeufiaceae from Thailand and China

Abstract

Tubeufiaceae, a cosmopolitan family with a worldwide distribution, is mostly reported as saprobic on decaying woody materials from both aquatic and terrestrial habitats. The family is commonly found as helicosporous hyphomycetes, while some are chlamydosporous and phragmosporous. In this study, thirteen helicosporous hyphomycetes were collected from Thailand and China. The phylogenetic analyses of combined ITS, LSU, TEF1-α, and RPB2 sequence data placed them in Dematiohelicomyces, Helicoma, Helicotruncatum, Neohelicosporium, Parahelicomyces, and Tubeufia within Tubeufiaceae. Three new species, Tubeufia cocois, Parahelicomyces chiangmaiensis, and Neohelicosporium bambusicola, one new host record, Tubeufia laxispora, and one new geographic record, T. longihelicospora, are introduced based on both morphological characteristics and phylogenetic analyses. In addition, Dematiohelicomyces helicosporus, Helicoma guttulatum, Helicotruncatum palmigenum, and Tubeufia cylindrothecia are described with detailed descriptions and color photo plates.

Full text

J. Fungi 2022, 8, 206. https://doi.org/10.3390/jof8020206 www.mdpi.com/journal/jof
Article
Three New Species, Two New Records and Four New
Collections of Tubeufiaceae from Thailand and China
Xingguo Tian
1,2,3,4,5
, Samantha C. Karunarathna
1,2,6
, Rongju Xu
4,5,7
, Yongzhong Lu
3
, Nakarin Suwannarach
8
,
Ausana Mapook
4
, Danfeng Bao
4,9
, Jianchu Xu
2,6,
* and Saowaluck Tibpromma
1,2,6,
*
1
Center for Yunnan Plateau Biological Resources Protection and Utilization, College of Biological Resource
and Food Engineering, Qujing Normal University, Qujing 655011, China;
6271105511@lamduan.mfu.ac.th (X.T.); samantha@mail.kib.ac.cn (S.C.K.)
2
Centre for Mountain Futures, Kunming Institute of Botany, Kunming 650201, China
3
School of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550003, China;
yzlu@git.edu.cn
4
Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand;
xurongju1005@outlook.com (R.X.); phung.ausana@gmail.com (A.M.); baodan_feng@cmu.ac.th (D.B.)
5
School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
6
CIFOR-ICRAF China Program, World Agroforestry (ICRAF), Kunming 650201, China
7
Yunnan Key Laboratory of Fungal Diversity and Green Development, Kunming Institute of Botany,
Kunming 650201, China
8
Research Center of Microbial Diversity and Sustainable Utilization, Faculty of Science,
Chiang Mai University, Chiang Mai 50200, Thailand; suwan_461@hotmail.com
9
College of Agriculture & Biological Science, Dali University, Dali 671003, China
* Correspondence: jxu@mail.kib.ac.cn (J.X.); saowaluckfai@gmail.com (S.T.)
Abstract: Tubeufiaceae, a cosmopolitan family with a worldwide distribution, is mostly reported as
saprobic on decaying woody materials from both aquatic and terrestrial habitats. The family is com-
monly found as helicosporous hyphomycetes, while some are chlamydosporous and phragmospo-
rous. In this study, thirteen helicosporous hyphomycetes were collected from Thailand and China.
The phylogenetic analyses of combined ITS, LSU, TEF1-α, and RPB2 sequence data placed them in
Dematiohelicomyces, Helicoma, Helicotruncatum, Neohelicosporium, Parahelicomyces, and Tubeufia within
Tubeufiaceae. Three new species, Tubeufia cocois, Parahelicomyces chiangmaiensis, and Neohelico-
sporium bambusicola, one new host record, Tubeufia laxispora, and one new geographic record, T.
longihelicospora, are introduced based on both morphological characteristics and phylogenetic anal-
yses. In addition, Dematiohelicomyces helicosporus, Helicoma guttulatum, Helicotruncatum palmigenum,
and Tubeufia cylindrothecia are described with detailed descriptions and color photo plates.
Keywords: three new species; two new records; asexual morph; phylogeny; taxonomy; Tubeufia lax-
ispora
1. Introduction
The order Tubeufiales was introduced by Boonmee et al. [1] to accommodate a single
family Tubeufiaceae based on phylogenic evidence. Later, the other two families, Bezerro-
mycetaceae and Wiesneriomycetaceae, were accepted into Tubeufiales by Liu et al. [2]
based on phylogenetic analysis and divergence time estimates. Tubeufiales currently in-
cludes three families viz. Bezerromycetaceae, Tubeufiaceae, and Wiesneriomycetaceae, of
which Bezerromycetaceae and Wiesneriomycetaceae are known for only sexual morphs,
while Tubeufiaceae has been reported to have both sexual and asexual morphs, and the
asexual morphs are mostly found as helicosporous hyphomycetes [2–4].
Tubeufiaceae was introduced by Barr [5] with Tubeufia as the generic type. The sexual
morph of Tubeufiaceae is characterized by superficial ascomata, pseudoparaphysate ha-
mathecium, and multiseptate, which are hyaline to pale brown cylindrical ascospores [5–
Citation: Tian, X.; Karunarathna,
S.C.; Xu, R.; Lu, Y.; Mapook, A.;
Suwannarach, N.; Bao, D.; Xu, J.;
Tibpromma, S. Three New Species,
Two New Records and Four New
Collections of Tubeufiaceae from
Thailand and China. J. Fungi 2022, 8,
206. https://doi.org/10.3390/
j
of8020206
Academic Editor: José Francisco
Cano-Lira
Received: 29 January 2022
Accepted: 16 February 2022
Published: 20 February 2022
Publisher’s Note: MDPI stays neu-
tral with regard to jurisdictional
claims in published maps and institu-
tional affiliations.
Copyright: © 2022 by the authors. Li-
censee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and con-
ditions of the Creative Commons At-
tribution (CC BY) license (https://cre-
ativecommons.org/licenses/by/4.0/).

J. Fungi 2022, 8, 206 2 of 32
10], while the asexual morph is hyphomycetous, mostly helicosporous, and some are chla-
mydosporous and phragmosporous [3,11,12]. Tubeufiaceae was recently revised by Lu et
al. [3], providing an updated multi-gene phylogenetic tree for Tubeufiales with 13 new
genera in this family. Currently, the family comprises 46 genera viz: Acanthohelicospora,
Acanthophiobolus, Acanthostigma, Acanthostigmina, Acanthotubeufia, Aquaphila, Artocarpomy-
ces, Berkleasmium, Bifrontia, Boerlagiomyces, Camporesiomyces, Chaetosphaerulina, Chlamydo-
tubeufia, Dematiohelicoma, Dematiohelicomyces, Dematiohelicosporum, Dematiotubeufia, Dicty-
ospora, Discotubeufia, Helicangiospora, Helicoarctatus, Helicodochium, Helicohyalinum, Heli-
coma, Helicomyces, Helicosporium, Helicotruncatum, Helicotubeufia, Kamalomyces, Kevinhydea,
Manoharachariella, Muripulchra, Neoacanthostigma, Neochlamydotubeufia, Neohelicoma, Neo-
helicomyces, Neohelicosporium, Neotubeufia, Pleurohelicosporium, Podonectria, Pseudohelicomy-
ces, Pseudohelicoon, Tamhinispora, Thaxteriella, Thaxteriellopsis, and Tubeufia [3,4].
Tubeufiaceae is cosmopolitan with a worldwide distribution in both tropical and temper-
ate regions [1,3,5,12].
Previously, the taxonomic placement of Tubeufiaceae was uncertain, thus it has been
discussed by several mycologists. First, Barr [5] accommodated Tubeufiaceae in Pleospo-
rales based on the generic type Tubeufia and later it was followed by various mycologists
[6,7,13–15]. Eriksson and Winka [16], and Eriksson [17,18] preferred to accommodate
Tubeufiaceae within the Dothideales, while Eriksson [19], as well as Lumbsch and Huhn-
dorf [20], preferred to accommodate Tubeufiaceae within Dothideomycetes and Chaeto-
thyriomycetes incertae sedis. Based on 28S rDNA sequence data, Kodsueb et al. [21] pre-
ferred to keep Tubeufiaceae in Pleosporales (as the natural placement), which also follows
the ordinal circumscription of Barr [6,7], Sivanesan [13], Rossman [22], Crane et al. [14],
and Kirk et al. [15]. Species of Tubeufiaceae are commonly reported as saprobes on woody
substrates or submerged decaying wood in terrestrial and aquatic habitats [1–3,10,12,23].
Several studies reported that Tubeufiaceae species are able to produce active secondary
metabolites, which have potential anti-fungal, anti-bacterial, anti-diabetic, and anti-cancer
properties [3,24–26]. Several Tubeufiaceae studies have been carried out based on descrip-
tions, illustrations, and phylogenetic evidence in Asia, especially in China, India, Japan,
and Thailand [1–4,12,27].
In this study, thirteen helicosporous hyphomycetes were collected from Thailand
and China. Phylogenetic analyses of combined ITS, LSU, TEF1-α, and RPB2 sequence data
place them in Dematiohelicomyces, Helicoma, Helicotruncatum, Neohelicosporium, Parahelico-
myces, and Tubeufia. Three new species, Tubeufia cocois, Parahelicomyces chiangmaiensis, and
Neohelicosporium bambusicola are introduced with morphological and phylogenic evidence.
One new host record, Tubeufia laxispora, from Cocos nucifera in Thailand, as well as one
new geographic record, T. longihelicospora, in China, are introduced. In addition, four
known species, Dematiohelicomyces helicosporus, Helicoma guttulatum, Helicotruncatum pal-
migenum, and Tubeufia cylindrothecia are also described. Full descriptions, color photo-
graphs, and a phylogenetic tree to show the placement of nine taxa are provided.
2. Materials and Methods
2.1. Sample Collection, Isolation, and Specimen Examination
Decaying wood, leaves, and culms were collected from Chiang Rai and Chiang Mai
Provinces of Thailand from May 2020 to March 2021 and Yunnan Province of China in
September 2021. Specimens were brought to a mycology laboratory for observation. Tian
et al. [28,29] and Senanayake et al. [30] were followed for the morphological study and
single spore isolation. Morphological characteristics were examined under a stereomicro-
scope (Motic SMZ-171, Wetzlar, Germany). Conidiomata were observed and photo-
graphed using a Nikon ECLIPSE Ni-U compound microscope connected with a Nikon
camera series DS-Ri2. Germinating conidia were transferred aseptically to a potato dex-
trose agar (PDA) medium, incubated at 28 °C for 2–4 weeks, and the morphological char-
acteristics of cultures were recorded.

J. Fungi 2022, 8, 206 3 of 32
Herbarium specimens were deposited at the herbarium of the Mae Fah Luang Uni-
versity (MFLU) and Kunming Institute of Botany (HKAS), while the living cultures were
deposited at Mae Fah Luang University Culture Collection (MFLUCC) and Kunming In-
stitute of Botany Culture Collection (KUMCC). Faces of Fungi and Index Fungorum num-
bers were registered as outlined in Index Fungorum [31] and Jayasiri et al. [32].
2.2. DNA Extraction, PCR Amplification, and Sequencing
Genomic DNA was extracted from two-week-old living pure cultures grown on PDA
using the Biospin Fungus Genomic DNA extraction Kit (BioFlux, Kun Ming, P.R. China)
following the manufacturer’s protocol. DNA was subjected to PCR amplification to am-
plify the genes ITS, LSU TEF1-α, and RPB2, while internal transcribed spacer (ITS) with
the primer pair of ITS4/ITS5 [33], the partial large subunit nuclear rDNA (LSU) with the
primer pair of LR0R/LR5 [34], the translation elongation factor 1-alpha gene (TEF1-α) with
the primer pair of EF1–983F/EF1–2218R [35], and RNA polymerase II second largest sub-
unit (RPB2) with the primer pair of RPB2–5f/7cR [36]. The PCR was carried out using the
method described by Tian et al. [29]. ITS, LSU, and TEF1-α amplification reactions were
set using the method described by Cai et al. [37] and Lu et al. [38]. RPB2 amplification
reaction was set using the method described by Lu et al. [3]. PCR products were checked
and purified in 1% agarose gels and were sequenced at TsingKe Biological Technology
(Kunming) Co., China.
2.3. Phylogenetic Analyses
The raw sequences (ITS, LSU, TEF1-α, and RPB2) were spliced using SeqMan and
subjected to BLAST in GenBank to find closely related taxa. Sequences of four genes
downloaded from NCBI GenBank are listed in Table 1. A single gene sequence alignment
was generated with MAFFT v.7.110 online application [39,40] and trimmed using trimAl
v 1.2 with the ‘gappyout’ option [41]. Multiple genes were concatenated by Sequence Ma-
trix. Multigene phylogenetic analyses of the concatenated genes were reconstructed from
maximum likelihood (ML) and Bayesian inference (BI) analyses. Maximum likelihood
was performed using the online RAxML-HPC on XSEDE tool on CIPRES under the GTR-
GAMMA substitution model and 1,000 bootstrap replicates [39,42,43]. Bayesian inference
analysis was performed using the MrBayes on XSEDE tool on CIPRES [43]. The best-fit
models were selected as GTR+I+G for ITS, LSU, TEF1-α, and RPB2 for the Bayesian pos-
terior probability analysis. Two parallel runs were conducted using the default settings,
six simultaneous Markov chains were run for 50,000,000 generations, and trees were sam-
pled every 500th generation. The alignment generated in this study was submitted to
TreeBASE (https://treebase.org/treebase-web/home.html, accessed on 10 January 2022)
under the submission number ID29068. Trees were visualized with FigTree v1.4.4, and
layouts were carried out with Adobe Illustrator CS5 v. 16.0.0.
Table 1. Names, culture collection accession numbers, and corresponding GenBank accession num-
bers of the fungal taxa used in this study.
Taxa Strain Numbers GenBank Accession Numbers
ITS LSU TEF1-
α
RPB2
A
quaphila albicans BCC 3543 DQ341096 DQ341101 – –
A
quaphila albicans MFLUCC 16–0010 KX454165 KX454166 KY117034 MF535255
Berkleasmium concinnum ILLS 80803 KY582485 – – –
Berkleasmium fusiforme MFLUCC 17–1979 MH558694 MH558821 MH550885 MH551008
Berkleasmium longisporum MFLUCC 17–1990 MH558697 MH558824 MH550888 MH551011
Botryosphaeria agavesT MFLUCC 10–0051 JX646790 JX646807 JX646855 –
Botryosphaeria dothidea CBS 115476 – NG_027577 – –
Chlamydotubeufia cylindricaT MFLUCC 16–1130 MH558702 MH558830 MH550893 MH551018
Chlamydotubeufia huaikangplaensisT MFLUCC10–0926 JN865210 JN865198 – –

J. Fungi 2022, 8, 206 4 of 32
Chlamydotubeufia krabiensisT MFLUCC 16–1134 KY678767 KY678759 KY792598 MF535261
Dematiohelicomyces helicosporusT MFLUCC 16–0213 KX454169 KX454170 KY117035 MF535258
Dematiohelicomyces helicosporus MFLUCC 16–0003 MH558703 MH558831 MH550894 MH551019
Dematiohelicomyces helicosporus MFLUCC 16–0007 MH558704 MH558832 MH550895 MH551020
Dematiohelicomyces helicosporus KUMCC 21–0473 OM331856 OL985958 OM355487 –
Dictyospora thailandica MFLUCC 16–0215 KY873628 KY873623 KY873287 –
Dictyospora thailandica MFLUCC 11–0512 KF301528 KF301536 – –
Helicoarctatus aquaticusT MFLUCC 17–1996 MH558707 MH558835 MH550898 MH551024
Helicodochium aquaticum MFLUCC 16–0008 MH558708 MH558836 MH550899 MH551025
Helicodochium aquaticumT MFLUCC 17–2016 MH558709 MH558837 MH550900 MH551026
Helicohyalinum aquaticumT MFLUCC 16–1131 KY873625 KY873620 KY873284 MF535257
Helicohyalinum infundibulumT MFLUCC 16–1133 MH558712 MH558840 MH550903 MH551029
Helicoma ambiens UAMH 10533 AY916451 AY856916 – –
Helicoma ambiens UAMH 10534 AY916450 AY856869 – –
Helicoma aquaticumT MFLUCC 17–2025 MH558713 MH558841 MH550904 MH551030
Helicoma brunneisporumT MFLUCC 17–1983 MH558714 MH558842 MH550905 MH551031
Helicoma dennisii NBRC 30667 AY916455 AY856897 – –
Helicoma fusiformeT MFLUCC 17–1981 MH558715 – MH550906 –
Helicoma guttulatumT MFLUCC 16–0022 KX454171 KX454172 MF535254 MH551032
H
elicoma guttulatum MFLUCC 21–0152 OL545456 OL606150 OL964521 OL964527
Helicoma hongkongense MFLUCC 17–2005 MH558716 MH558843 MH550907 MH551033
Helicoma inthanonenseT MFLUCC 11–0003 JN865211 JN865199 – –
Helicoma khunkornensisT MFLUCC 10–0119 JN865203 JN865191 KF301559 _
Helicoma linderi NBRC 9207 AY916454 AY856895 – –
Helicoma longisporum MFLUCC 16–0002 MH558717 MH558844 MH550908 MH551034
Helicoma longisporum MFLUCC 16–0005 MH558718 – MH550909 MH551035
Helicoma longisporum MFLUCC 16–0211 MH558719 MH558845 MH550910 MH551036
Helicoma longisporumT MFLUCC 17–1997 MH558720 MH558846 MH550911 MH551037
Helicoma miscanthiT MFLUCC 11–0375 KF301525 KF301533 KF301554 –
Helicoma muelleri CBS 964.69 AY916453 AY856877 – –
Helicoma muelleri UBC F13877 AY916452 AY856917 – –
Helicoma multiseptatumT GZCC 16–0080 MH558721 MH558847 MH550912 MH551038
Helicoma nematosporumT MFLUCC 16–0011 MH558722 MH558848 MH550913 MH551039
Helicoma rubriappendiculatumT MFLUCC 18–0491 MH558723 MH558849 MH550914 MH551040
Helicoma rufumT MFLUCC 17–1806 MH558724 MH558850 MH550915 –
Helicoma rugosum ANM 196 GQ856138 GQ850482 – –
Helicoma rugosum ANM 953 GQ856139 GQ850483 – –
Helicoma rugosum ANM 1169 – GQ850484 – –
Helicoma rugosum
J
CM 2739 – AY856888 – –
Helicoma septoconstrictum MFLUCC 17–1991 MH558725 MH558851 MH550916 MH551041
Helicoma septoconstrictumT MFLUCC 17–2001 MH558726 MH558852 MH550917 MH551042
Helicoma siamenseT MFLUCC 10–0120 JN865204 JN865192 KF301558 _
Helicoma sp. HKUCC 9118 – AY849966 – –
Helicoma tectonaeT MFLUCC 12–0563 KU144928 KU764713 KU872751 _
Helicoma vaccinii CBS 216.90 AY916486 AY856879 _ _
Helicomyces chiayiensisT BCRC FU30842 LC316604 – – –
Helicomyces colligatus MFLUCC 16–1132 MH558727 MH558853 MH550918 MH551043
Helicomyces hyalosporus GZCC 16–0070 MH558728 MH558854 MH550919 MH551044
Helicomyces hyalosporus MFLUCC 17–0051 MH558731 MH558857 MH550922 MH551047
Helicomyces torquatus MFLUCC 16–0217 MH558732 MH558858 MH550923 MH551048
Helicosporium flavumT MFLUCC 16–1230 KY873626 KY873621 KY873285 –

J. Fungi 2022, 8, 206 5 of 32
Helicosporium luteosporumT MFLUCC 16–0226 KY321324 KY321327 KY792601 MH551056
Helicosporium vesicariumT MFLUCC 17–1795 MH558739 MH558864 MH550930 MH551055
Helicotruncatum palmigenum NBRC 32663 AY916480 AY856898 – –
Helicotruncatum palmigenum MFLUCC 15–0993 MT627685 MN913690 – –
H
elicotruncatum palmigenum KUMCC 21–0474 OM102542 OL985959 OM355488 OM355492
Helicotubeufia guangxiensisT MFLUCC 17–0040 MH290018 MH290023 MH290028 MH290033
Helicotubeufia hydeiT MFLUCC 17–1980 MH290021 MH290026 MH290031 MH290036
Helicotubeufia jonesiiT MFLUCC 17–0043 MH290020 MH290025 MH290030 MH290035
M
uripulchra aquatica DLUCC 0571 KY320531 KY320548 – –
M
uripulchra aquatica KUMCC 15–0245 KY320533 KY320550 KY320563 MH551057
M
uripulchra aquatica KUMCC 15–0276 KY320534 KY320551 KY320564 MH551058
M
uripulchra aquaticaT MFLUCC 15–0249 KY320532 KY320549 – –
Neoacanthostigma fusiformeT MFLUCC 11–0510 KF301529 KF301537 – –
Neochlamydotubeufia fusiformisT MFLUCC 16–0016 MH558740 MH558865 MH550931 MH551059
Neochlamydotubeufia fusiformis MFLUCC 16–0214 MH558741 MH558866 MH550932 MH551060
Neochlamydotubeufia khunkornensisT MFLUCC 10–0118 JN865202 JN865190 KF301564 –
Neochlamydotubeufia khunkornensis MFLUCC 16–0025 MH558742 MH558867 MH550933 MH551061
Neohelicomyces aquaticus KUMCC 15–0463 KY320529 KY320546 KY320562 MH551065
Neohelicomyces grandisporusT KUMCC 15–0470 KX454173 KX454174 – MH551067
Neohelicomyces submersusT MFLUCC 16–1106 KY320530 KY320547 – MH551068
Neohelicosporium abuense CBS 101688 AY916470 – – –
Neohelicosporium acrogenisporumT MFLUCC 17–2019 MH558746 MH558871 MH550937 MH551069
Neohelicosporium aquaticumT MFLUCC 17–1519 MF467916 MF467929 MF535242 MF535272
Neohelicosporium astrictumT MFLUCC 17–2004 MH558747 MH558872 MH550938 MH551070
Neohelicosporium aurantiellum ANM 718 GQ856140 GQ850485 – –
Neohelicosporium bambusicolaT MFLUCC 21–0156 OL606157 OL606146 OL964517 OL964523
Neohelicosporium ellipsoideumT MFLUCC 16–0229 MH558748 MH558873 MH550939 MH551071
Neohelicosporium fusisporumT MFUCC 16–0642 MG017612 MG017613 MG017614 –
Neohelicosporium griseum CBS 961.69 AY916474 AY856884 – –
Neohelicosporium griseum CBS 113542 AY916475 AY916088 – –
Neohelicosporium guangxiense GZCC 16–0042 MF467920 MF467933 MF535246 MF535276
Neohelicosporium guangxiense MFLUCC 17–0054 MH558750 MH558875 MH550941 MH551073
Neohelicosporium hyalosporum GZCC 16–0063 MH558751 MH558876 MH550942 MH551074
Neohelicosporium hyalosporumT GZCC 16–0076 MF467923 MF467936 MF535249 MF535279
Neohelicosporium irregulareT MFLUCC 17–1796 MH558752 MH558877 MH550943 MH551075
Neohelicosporium irregulare MFLUCC 17–1808 MH558753 MH558878 MH550944 MH551076
Neohelicosporium krabienseT MFLUCC 16–0224 MH558754 MH558879 MH550945 MH551077
Neohelicosporium laxisporumT MFLUCC 17–2027 MH558755 MH558880 MH550946 MH551078
Neohelicosporium morganii CBS 281.54 AY916468 AY856876 – –
Neohelicosporium morganii CBS 222.58 AY916469 AY856880 – –
Neohelicosporium ovoideumT GZCC 16–0064 MH558756 MH558881 MH550947 MH551079
Neohelicosporium ovoideum GZCC 16–0066 MH558757 MH558882 MH550948 MH551080
Neohelicosporium panacheum CBS 257.59 AY916471 AY916087 – –
Neohelicosporium parvisporum GZCC 16–0078 MF467924 MF467937 MF535250 MF535280
Neohelicosporium parvisporum MFLUCC 17–2010 MH558763 MH558888 MH550954 MH551086
Neohelicosporium sp. CBS 189.95 AY916472 AY856882 – –
Neohelicosporium sp. HKUCC 10235 – AY849942 – –
Neohelicosporium submersum MFLUCC 17–2376 MT627738 MN913738 – –
Neohelicosporium taiwanenseT BCRC FU30841 LC316603 – – –
Neohelicosporium thailandicumT MFLUCC 16–0221 MF467928 MF467941 MF535253 MF535283
Neotubeufia krabiensisT MFLUCC 16–1125 MG012031 MG012024 MG012010 MG012017

J. Fungi 2022, 8, 206 6 of 32
Parahelicomyces aquaticusT MFLUCC 16–0234 MH558766 MH558891 MH550958 MH551092
Parahelicomyces chiangmaiensisT MFLUCC 21–0159 OL697884 OL606145 OL964516 OL964522
Parahelicomyces hyalosporus CBS 283.51 AY916464 AY856881 DQ677928 DQ677981
Parahelicomyces hyalosporus KUMCC 15–0411 KY320527 KY320544 KY320560 –
Parahelicomyces hyalosporusT MFLUCC 15–0343 KY320523 KY320540 – –
Parahelicomyces indicus CBS 374.93 AY916477 AY856885 – –
Parahelicomyces menglunicusT HKAS 85793 MK335914 – MK335916
Parahelicomyces paludosus CBS 120503 DQ341095 DQ341103 – –
Parahelicomyces quercus MFLU 18-2091 – – MK360077 MK434906
Parahelicomyces quercus MFUCC 17-0895 MK347720 MK347934 – –
Parahelicomyces talbotii MUCL 33010 AY916465 AY856874 – –
Parahelicomyces talbotii MFLUCC 17–2021 MH558765 MH558890 MH550957 MH551091
Pseudohelicoon gigantisporum BCC 3550 AY916467 AY856904 – –
Pseudohelicoon subglobosumT BCRC FU30843 LC316607 LC316610 – –
Thaxteriellopsis lignicola MFLUCC 10–0123 JN865207 JN865195 KF301562 _
Thaxteriellopsis lignicola MFLUCC 10–0124 JN865208 JN865196 KF301561 _
Tubeufia abundataT MFLUCC 17–2024 MH558769 MH558894 MH550961 MH551095
Tubeufia aquatica MFLUCC 17–1794 MH558770 MH558895 MH550962 MH551096
Tubeufia aquaticaT MFLUCC 16–1249 KY320522 KY320539 KY320556 MH551142
Tubeufia aquatica DLUCC 0574 KY320521 KY320538 KY320555 MH551141
Tubeufia bambusicolaT MFLUCC 17–1803 MH558771 MH558896 MH550963 MH551097
Tubeufia brevisT MFLUCC 17–1799 MH558772 MH558897 MH550964 MH551098
Tubeufia brunneaT MFLUCC 17–2022 MH558773 MH558898 MH550965 MH551099
Tubeufia chiangmaiensis MFLUCC 17–1801 MH558774 MH558899 MH550966 MH551100
Tubeufia chiangmaiensisT MFLUCC 11–0514 KF301530 KF301538 KF301557 –
Tubeufia chlamydosporaT MFLUCC 16–0223 MH558775 MH558900 MH550967 MH551101
Tubeufia cocoisT MFLUCC 22–0001 OM102541 OL985957 OM355486 OM355491
Tubeufia cocois MFLUCC 22–0002 OM102543 OL985960 OM355489 OM355493
Tubeufia cocois MFLUCC 22–0003 OM102544 OL985961 OM355490 OM355494
Tubeufia cylindrothecia BCC 3559 – AY849965 – –
Tubeufia cylindrothecia BCC 3585 AY916482 AY856908 – –
Tubeufia cylindrothecia DLUCC 0572 KY320520 KY320537 KY320554 –
Tubeufia cylindrothecia MFLUCC 16–1253 KY320519 KY320536 KY320553 –
Tubeufia cylindrothecia MFLUCC 16–1283 KY320518 KY320535 KY320552 MH551143
Tubeufia cylindrothecia MFLUCC 21–0160 OL545365 OL606147 OL964518 OL964524
Tubeufia cylindrothecia MFLUCC 17–1792 MH558776 MH558901 MH550968 MH551102
Tubeufia dictyosporaT MFLUCC 17–1805 MH558778 MH558903 MH550970 –
Tubeufia dictyospora MFLUCC 16–0220 MH558777 MH558902 MH550969 MH551103
Tubeufia eccentricaT MFLUCC 17–1524 MH558782 MH558907 MH550974 MH551108
Tubeufia eccentrica GZCC 16–0035 MH558779 MH558904 MH550971 MH551105
Tubeufia entadae MFLU 18–2102 NR_163323 – – –
Tubeufia entadae MFLU 18-2102 MK347727 MK347943 – –
Tubeufia fangchengensisT MFLUCC 17–0047 MH558783 MH558908 MH550975 MH551109
Tubeufia filiformisT MFLUCC 16–1128 – KY092407 KY117028 MF535284
Tubeufia filiformis MFLUCC 16–1135 KY092416 KY092411 KY117032 MF535285
Tubeufia filiformis MFLUCC 16–0236 – MH558938 MH550976 MH551110
Tubeufia geniculataT BCRC FU30849 LC335817 – – –
Tubeufia geniculata NCYU U2–1B LC335816 – – –
Tubeufia guangxiensis MFLUCC 17–0046 MH558784 MH558909 MH550977 MH551111
Tubeufia hechiensisT MFLUCC 17–0052 MH558785 MH558910 MH550978 MH551112
Tubeufia hyalosporaT MFLUCC 15–1250 MH558786 MH558911 MH550979 –

J. Fungi 2022, 8, 206 7 of 32
Tubeufia inaequalis GZCC 16–0079 MH558787 MH558912 MH550980 MH551113
Tubeufia inaequalis MFLUCC 17–1998 MH558791 MH558916 MH550984 MH551117
Tubeufia inaequalis BCC 8808 AY916481 AY856910 – –
Tubeufia javanica MFLUCC 12–0545 KJ880034 KJ880036 KJ880037 –
Tubeufia krabiensisT MFLUCC 16–0228 MH558792 MH558917 MH550985 MH551118
Tubeufia latisporaT MFLUCC 16–0027 KY092417 KY092412 KY117033 MH551119
Tubeufia laxispora MFLUCC 16–0013 MH558793 MH558918 MH550986 MH551120
Tubeufia laxispora MFLUCC 16–0219 KY092414 KY092409 KY117030 MF535286
Tubeufia laxisporaT MFLUCC 16–0232 KY092413 KY092408 KY117029 MF535287
Tubeufia laxispora MFLUCC 17–2023 MH558794 MH558919 MH550987 MH551121
Tubeufia laxispora MFLUCC 21–0163 OL545455 OL606148 OL964519 OL964525
Tubeufia lilliputea NBRC 32664 AY916483 AY856899 – –
Tubeufia longihelicosporaT MFLUCC 16–0753 MZ538531 MZ538565 MZ567106 –
Tubeufia longihelicospora MFLUCC 21–0151 OL606156 OL606149 OL964520 OL964526
Tubeufia longihelicospora KUMCC 21–0814 OM331690 OM331688 OM355484 –
Tubeufia longihelicospora KUMCC 21–0815 OM331691 OM331705 OM355485 –
Tubeufia longisetaT MFLUCC 15–0188 KU940133 – – –
Tubeufia mackenzieiT MFLUCC 16–0222 KY092415 KY092410 KY117031 MF535288
Tubeufia parvispora MFLUCC 17–1992 MH558796 MH558921 MH550989 MH551123
Tubeufia parvispora MFLUCC 17–2003 MH558797 MH558922 MH550990 MH551124
Tubeufia parvispora MFLUCC 17–2009 MH558798 MH558923 MH550991 MH551125
Tubeufia roseohelicospora MFLUCC 16–0230 MH558799 MH558924 MH550992 MH551126
Tubeufia roseohelicospora MFLUCC 17–1797 MH558800 MH558925 MH550993 MH551127
Tubeufia roseohelicosporaT MFLUCC 15–1247 KX454177 KX454178 – MH551144
Tubeufia rubra GZCC 16–0083 MH558802 MH558927 MH550995 MH551129
Tubeufia rubraT GZCC 16–0081 MH558801 MH558926 MH550994 MH551128
Tubeufia sahyadriensisT NFCCI 4252 MH033849 MH033850 MH033851 –
Tubeufia sessilisT MFLUCC 16–0021 MH558803 – MH550996 MH551130
Tubeufia sympodihylosporaT MFLUCC 17–0044 MH558806 MH558930 MH550999 MH551133
Tubeufia sympodilaxispora BCC 3580 – DQ296554 – –
Tubeufia sympodilaxispora GZCC 16–0058 MH558807 MH558931 MH551000 MH551134
Tubeufia sympodilaxisporaT MFLUCC 17–0048 MH558808 MH558932 MH551001 MH551135
Tubeufia taiwanensisT BCRC FU30844 LC316605 – – –
Tubeufia tectonae MFLUCC 16–0235 MH558809 MH558933 MH551002 MH551136
Tubeufia tectonae MFLUCC 17–1985 MH558810 MH558934 MH551003 MH551137
Tubeufia tectonaeT MFLUCC 12–0392 KU144923 KU764706 KU872763 –
Tubeufia tratensisT MFLUCC 17–1993 MH558811 MH558935 MH551004 MH551138
Tubeufia xylophila MFLUCC 17–1520 MH558813 MH558937 MH551006 MH551140
Tubeufia xylophila GZCC 16–0038 MH558812 MH558936 MH551005 MH551139
Notes: Ex-type strains are indicated by T after the species name. Newly generated sequences are in
black bold. The symbol “–” indicates information not available. Abbreviations: ANM, A.N. Miller;
BCC, Biotec Culture Collection, Thailand; BCRC, Bioresearch Collection and Research Centre; CBS,
Westerdijk Fungal Biodiversity Institute; DLUCC, Culture collection of Dali University; GUCC, Gui-
zhou University Culture Collection; HKAS, the herbarium of Cryptogams Kunming Institute of
Botany Academia Sinica; HKUCC, Hong Kong University Culture Collection; JCM, Japan Collection
of Microorganisms; KUMCC, Culture collection of Kunming Institute of Botany; MFLU, the herbar-
ium of the Mae Fah Luang University; MFLUCC, Mae Fah Luang University Culture Collection;
NBRC, NITE Biological Resource Center; NCYU, National Chiayi University; NFCCI, National Fun-
gal Culture Collection of India; UAMH, the University of Alberta Microfungus Collection and Her-
barium; UBC F, University of British Columbia Herbarium.

J. Fungi 2022, 8, 206 8 of 32
3. Results
3.1. Phylogenetic Analyses
The combined ITS, LSU, TEF1-α, and RPB2 dataset comprised thirteen newly se-
quenced strains, with Botryosphaeria dothidea (CBS 115476) and B. agaves (MFLUCC 10–
0051) as outgroup taxa. Multiple genes were concatenated, which comprised 3425 nucle-
otide characters, including gaps (ITS: 1–606 bp, LSU: 607–1471 bp, RPB2: 1472–2514 bp,
TEF1-α: 2515–3425 bp). The RAxML analysis of the combined dataset yielded the best-
scoring tree (Figure 1) with a final ML optimization likelihood value of -50085.613741. The
matrix had 1671 distinct alignment patterns, with 26.72% undetermined characters or
gaps. Estimated base frequencies were as follows: A = 0.244864, C = 0.251768, G = 0.258885,
T = 0.244483; substitution rates AC = 1.177463, AG = 5.880242, AT = 2.143397, CG =
0.867985, CT = 9.022451, GT = 1.000000; gamma distribution shape parameter α = 0.224491.
Phylogenetic analyses showed that our thirteen collections were placed within
Tubeufiaceae viz: Dematiohelicomyces, Helicoma, Helicotruncatum, Neohelicosporium, Para-
helicomyces, and Tubeufia. Eight collections clustered within Tubeufia; the new strain
Tubeufia cylindrothecia (MFLUCC 21–0160) was nested with six strains of T. cylindrothecia
with strong bootstrap support (98% ML/1.00 PP). Three strains of Tubeufia cocois
(MFLUCC 22–0001, MFLUCC 22–0002, and MFLUCC 22–0003) clustered together and
formed a branch at the basal clades of T. aquatica with strong bootstrap support (100%
ML/1.00 PP). The new strain T. laxispora (MFLUCC 21–0163) nested with four strains of T.
laxispora with strong bootstrap support (100% ML/1.00 PP), and T. longihelicospora
(MFLUCC 21–0151, KUMCC 21–0478, and KUMCC 21–0479) clustered together within
the same clade as T. longihelicospora (MFLUCC 16–0753). Parahelicomyces chiangmaiense
(MFLUCC 21–0159) formed a single branch at the basal clades of Parahelicomyces members
with strong support (97% ML). Newly obtained strain Helicotruncatum palmigenum
(KUMCC 21–0474) nested with two strains of H. palmigenum (NMRC 32,663 and MFLUCC
15–0093) strong bootstrap support (100% ML/1.00 PP). Neohelicosporium bambusicola
(MFLUCC 21–0160) was placed as a sister taxon to N. ellipsoideum (MFLUCC 16–0229) and
N. acrogenisporum (MFLUCC 17–2019). New strain Dematiohelicomyces helicosporus
(KUMCC 21–0473) clustered with three strains of Dematiohelicomyces helicosporus with
strong bootstrap support (100% ML/1.00 PP), and Helicoma guttulatum (MFLUCC 21–0152)
clustered with its ex-type strain of H. guttulatum (MFLUCC 16–0022) with high support
(100% ML/1.00 PP).

J. Fungi 2022, 8, 206 9 of 32

J. Fungi 2022, 8, 206 10 of 32

J. Fungi 2022, 8, 206 11 of 32
Figure 1. Phylogenetic tree generated from a maximum likelihood analysis based on a concatenated
alignment of ITS, LSU, TEF1-α, and RPB2 sequences data in Tubeufiaceae. The tree is rooted with
Botryosphaeria dothidea (CBS 115476) and B. agaves (MFLUCC 10–0051). Bootstrap support values
equal to or higher than 75% ML (left) or posterior probability values equal to or higher than 0.95
Bayesian PP (right) are indicated on the nodes. Newly generated sequences are in red. Ex-type
strains are in black/red bold.
3.2. Taxonomy
3.2.1. Dematiohelicomyces Y.Z. Lu, Boonmee, and K.D. Hyde, Fungal Diversity 92: 159
(2018)
Index Fungorum, IF 554824; Facesoffungi number, FoF 04701
Type species: Dematiohelicomyces helicosporus (Boonmee, Y.Z. Lu, and K.D. Hyde) Y.Z.
Lu
The monotypic genus Dematiohelicomyces was introduced by Lu et al. [3], with D. hel-
icosporus as the type species based on morphology and phylogeny. Dematiohelicomyces are
saprobic on submerged decaying wood in a freshwater stream in Thailand. Dematiohelico-
myces is characterized by short conidiophores that are brown, 0–3-septate, and helicoid
conidia, with a spathulate basal end cell. In this paper, Dematiohelicomyces helicosporus was
collected from submerged decaying wood in a freshwater river in Thailand.
Dematiohelicomyces helicosporus (Boonmee, Y.Z. Lu, and K.D. Hyde) Y.Z. Lu, Fungal
Diversity 92: 159 (2018) (Figure 2).
≡ Chlamydotubeufia helicospora Boonmee, Y.Z. Lu, and K.D. Hyde, Fungal Diversity 80:
123 (2016)
Index Fungorum, IF 554825; Facesoffungi number, FoF 04702

J. Fungi 2022, 8, 206 12 of 32
Saprobic on submerged decaying wood in a freshwater stream. Sexual morph Unde-
termined. Asexual morph Hyphomycetous, helicosporous. Colonies are superficial, effuse,
gregarious, white, and shiny. Mycelium is mostly immersed, composed of branched, sep-
tate hyphae, brown, with masses of glistening, crowded conidia. Conidiophores (16.5–)30–
65.5(–80.5) × 4–5 μm (x = 47 × 4.5 μm, n = 25) are macronematous, erect, cylindrical,
branched, 0–4-septate, hyaline to pale brown, arising as lateral branches from creeping
hyphae, and smooth-walled. Conidiogenous cells (9–)14–24.5(–30) × 4–5 μm (x = 19.5 × 4.5
μm, n = 30) are holoblastic, monoblastic, integrated, terminal, cylindrical, truncate at the
apex after conidial secession, hyaline, and smooth-walled. Conidia are solitary, acroge-
nous, helicoid, rounded at tip, with the basal cells broadly spathulate and bearing a flat-
tened attachment scar, guttulate, hyaline, with a (59–)80–139(–158) μm (x = 110 μm, n =
25) diam. and conidial filament 5–6.5 μm (x = 5.5 μm, n = 25) wide in the broadest part,
tapering towards the ends, 328–482.5 μm (x = 405.5 μm, n = 25) long, multi-septate, coiled
1–2 ½times, tightly to loosely coiled in water, smooth-walled, and contain granules.
Culture characteristics: conidia germinating on PDA within 12 h; colonies growing
on PDA, reaching 20 mm in 2 weeks at 28 °C, circular, with a flat surface, edge entire, pale
brown to dark brown in PDA medium; mycelium partially immersed, branched, multi-
septate, hyaline to pale brown, smooth.
Material examined: Thailand, Chiang Rai Province, Mae Fah Luang University, on
submerged decaying wood, 22 May 2020, R. J. Xu, MD38 (MFLU 21–0184), living culture,
KUMCC 21–0473.
Notes: There are some differences between our new isolate (KUMCC 21–0473) and
D. helicosporus morphologically, such as conidiogenous cells in our new isolate are mono-
blastic, while in D. helicosporus (MFLUCC 16–0003), they are mono- to polyblastic. In ad-
dition, the new isolate (KUMCC 21–0473) differs from D. helicosporus (MFLUCC 16–0003)
in having larger (80–139 vs. 70–100 μm) and shorter conidial filaments (328–482.5 vs. 400–
600 μm) [3,44]. However, our phylogenetic results show that the new isolate D. helicospo-
rus (KUMCC 21–0473) clusters with three strains of D. helicosporus (MFLUCC 16–0003,
MFLUCC 16–0007, and MFLUCC 16–0213) with high statistical supports (100% ML/1.00
PP, Figure 1). Therefore, we identify our new isolate as Dematiohelicomyces helicosporus.
Chlamydotubeufia helicospora was collected on decaying wood in a flowing freshwater
stream in Uttaradit Province, Thailand [44], and later, based on morphology and phylog-
eny, Lu et al. [3] synonymized this taxon under Dematiohelicomyces helicosporus. In this
study, our new isolate was also collected from a submerged decaying wood in Chiang Rai
Province, Thailand, which is a little far from the original collection location, meaning that
this species still prefers similar environmental conditions.

J. Fungi 2022, 8, 206 13 of 32
Figure 2. Dematiohelicomyces helicosporus (MFLU 21–0184). (a,b) Colony on decaying wood; (c,d)
conidiophores and conidia; (e–g) conidiophores and conidiogenous cells; (h–l) conidia; (m) germi-
nated conidium; (n,o) colony cultures on PDA (observe and reverse). Scale bars: (j) = 80 μm, (d) =
60 μm, (c,h,i,k–m) = 40 μm, (e–g) = 20 μm.
3.2.2. Helicoma Corda, Icon. fung. (Prague) 1: 15 (1837)
Index Fungorum: IF 8473
Type species: Helicoma muelleri Corda, Icon.
Helicoma was introduced by Corda [45], with H. muelleri as a type species. Two types
of asexual morphs have been observed in Helicoma: the first asexual morphs are charac-
terized by conidiogenous cells that are cylindrical, with denticles, intercalary, arising lat-
erally from the lower portion of conidiophores, and conidia are pleurogenous, tapering

J. Fungi 2022, 8, 206 14 of 32
towards the apex and rounded at the tip, helicoid, hygroscopic, and become loosely coiled
in water [12]. Another asexual morph is characterized by conidia that are acrogenous or
acropleurogenous, helicoid, circinate, dry, tapering towards the apex, truncating at the
base, coiled 1¼–¾ times, and not becoming loose in the water. There are 97 records listed
in Index Fungorum (2021), however, most of them are lacking sequence data in GenBank.
The last treatment of Helicoma was provided by Lu et al. [3], and they accepted 57 species
within the genus while introducing 10 new species and 11 new combinations. In this
study, the new isolate is identified as H. guttulatum based on both phylogenetic analysis
and morphological characteristics.
Helicoma guttulatum Y.Z. Lu, Boonmee, and K.D. Hyde, Fungal Diversity 80: 125 (2016)
(Figure 3).
Index Fungorum, IF 552218; Facesoffungi, FoF 02358
Saprobic on submerged decaying wood in a freshwater stream. Sexual morph Unde-
termined. Asexual morph Hyphomycetous, helicosporous. Colonies are superficial, effuse,
gregarious, brown to dark brown. Mycelium is mostly immersed, partly superficial, com-
posed of branched, septate, brown hyphae. Conidiophores (65–)93–156.5 × 4.5–6 μm (x = 125
× 5 μm, n = 20) are macronematous, mononematous, cylindrical, septate, erect, un-
branched, pale brown to brown at the apex and dark brown at the base, and smooth-
walled. Conidiogenous cells (9–)12–24(–30.5) × 4–5.5 μm (x = 18 × 4.5 μm, n = 20) are holo-
blastic, mono- to polyblastic, integrated, terminal, cylindrical, brown, and smooth-walled.
Conidia 22–26.5 μm (x = 24 μm, n = 25) have a diam. and conidial filament 7–8.5 μm (x = 8
μm, n = 25) wide and 49–58 μm (x = 53.5 μm, n = 25) long, are integrated, terminal, helicoid,
tightly coiled 1–1½ times, guttulate, do not become loose in the water, 8-septate, straight
constricted at the septa, subhyaline to yellowish, rounded at the apex, and smooth-walled.
Culture characteristics: conidia germinated on PDA within 12 h; colonies growing on
PDA, reaching 25 mm in 2 weeks at 28 °C, circular, with a flat surface, edge entire, and
pale brown to brown in PDA medium; mycelium were partially immersed, branched,
multi-septate, hyaline to pale brown, and smooth.
Material examined: Thailand, Chiang Rai Province, Mueang, Nang Lae on sub-
merged decaying wood, 14 August 2020, R. J. Xu, MD106 (MFLU 21–0183), living culture,
MFLUCC 21–0152.
Notes: Helicoma guttulatum was introduced by Hyde et al. [44] on submerged decay-
ing wood from a freshwater stream in Thailand. In our phylogenetic analyses, the newly
obtained isolate (MFLUCC 21–0152) clustered with the ex-type strain of H. guttulatum
(MFLUCC 16–0022) with high statistical support (100% ML/1.00 PP, Figure 1). Morpho-
logically, the new isolate was indistinguishable from the holotype of H. guttulatum [44].
Therefore, we identify the new isolate as Helicoma guttulatum based on morphological and
phylogenetic data.

J. Fungi 2022, 8, 206 15 of 32
Figure 3. Helicoma guttulatum (MFLU 21–0183). (a) Colony on decaying wood; (b,c) conidiophores
and conidia; (d) conidiophore; (e,f) tip of conidiogenous cells; (g,h) conidiogenous cells and conidia;
(i–k) conidia; (l) germinated conidium; (m,n) colony cultures on PDA (observe and reverse). Scale
bars: (b) = 40 μm, (c,d) = 60 μm, (e–l) = 20 μm.
3.2.3. Helicotruncatum Y.Z. Lu, J.C. Kang, and K.D. Hyde, Fungal Diversity 92: 220 (2018)
Index Fungorum, IF 554859; Facesoffungi number, FoF 04,730
Type species: Helicotruncatum palmigenum (Penz. and Sacc.) Y.Z. Lu and K.D. Hyde
The monotypic genus Helicotruncatum was established by Lu et al. [3] with H. pal-
migenum as the type species, and it is the only species accepted in the genus [3,12].

J. Fungi 2022, 8, 206 16 of 32
Helicotruncatum palmigenum was originally placed in Helicoma, based on morphological
characters [12,46,47]. Phylogenetic analysis of Lu et al. [3] showed that H. palmigenum
formed an independent lineage and was distant from Helicoma. Morphologically, H. pal-
migenum can be distinguished from other helicosporous hyphomycetes by the distinc-
tively thickened lateral cell wall of the conidiophore and basal cell of the conidium. Thus,
Lu et al. [3] introduced a new genus, Helicotruncatum, to accommodate H. palmigenum
based on both phylogeny and morphology. In this paper, Helicotruncatum palmigenum was
collected from dead Cocos nucifera leaves in Thailand.
Helicotruncatum palmigenum (Penz. and Sacc.) Y.Z. Lu and K.D. Hyde, Fungal Diversity
92: 220 (2018) (Figure 4).
≡Helicosporium intermedium var. palmigenum Penz. and Sacc., Malpighia 15(7–9): 249
(1902)
≡Helicoma palmigenum (Penz. and Sacc.) Linder, Ann. Mo. bot. Gdn 16: 306 (1929)
=Helicoma westonii Linder [as ‘westoni’], Ann. Mo. bot. Gdn 18: 12 (1931)
Index Fungorum: IF 554860; Facesoffungi number: FoF 04800
Saprobic on dead leaves of Cocos nucifera. Sexual morph: Undetermined. Asexual
morph Hyphomycetous, helicosporous. Colonies on the substratum are superficial, effuse,
gregarious, and velvety black. Mycelium is composed of brown, septate hyphae. Conidio-
phores 165.5–283.5 × 6.5–10 μm (x = 224.5 × 8.5 μm, n =10) are macronematous, mononem-
atous, cylindrical, stout, septate, erect, unbranched, pale brown to subhyaline at the apex
and dark brown at the base, and smooth-walled. Conidiogenous cells 22–34.5 × 6–7.5 μm (x
= 28 × 6.5 μm, n =15) are holoblastic, monoblastic, integrated, determinate, cylindrical,
terminal, smooth-walled, and truncate at the apex after conidial secession. Conidia 32–44
μm (x = 38 μm, n = 20) and conidial filament are 10.5–15 μm (x = 12.5 μm, n = 20) wide,
82–108.5 μm (x = 95.5 μm, n = 20) long, solitary, terminal, smooth-walled, helicoid, coiled
1½–2¾ times, do not become loose in the water, septate, not constricted at septa, dilute
fuliginously, and the basal cell is truncated with thickened lateral walls.
Culture characteristics: conidia germinating on PDA within 12 h; colonies reaching
40 mm in 2 weeks at 28 °C, irregular, dark brown from above and pale brown from below;
mycelium are slow-growing, thin, and effuse brownish grey.
Material examined: Thailand, Chiang Rai Province, on decaying leaves of Cocos nu-
cifera, 16 January 2021, X. G. Tian, C6–6, (MFLU 21–0185), living culture, KUMCC 21–0474.
Notes: In the phylogenetic analyses, our new collection KUMCC 21–0474 clusters
with two strains of H. palmigenum (NBRC 32663, MFLUCC 15–0993) with high statistical
supports (100% ML/1.00 PP, Figure 1). Morphologically, our new isolate is almost identi-
cal to H. palmigenum except for the size of the conidiogenous cells (22–34.5 vs. 17–25 μm
long) and the conidia (82–108.5 vs. 50–60 μm long). The nucleotide comparisons show 2
bp and 1 bp of ITS and LSU differences between the new isolate (KUMCC 21–0474) and
H. palmigenum (NBRC 32663). Thus, we identify the new isolate as H. palmigenum based
on both phylogenetic analyses and morphological characteristics.
Helicotruncatum palmigenum was introduced as Helicoma palmigenum by Linder [46]
on decaying petioles of palms that were collected from Australia, Brazil, China, Indonesia,
Japan, Mexico, New Guinea, Seychelles, Thailand, Trinidad, and the USA [3,46,47]. In ad-
dition, Helicotruncatum palmigenum has been reported on leaves and husks of Cocos nucifera
[35]. Our new isolate was also collected on dead leaves of Cocos nucifera from Thailand.

J. Fungi 2022, 8, 206 17 of 32
Figure 4. Helicotruncatum palmigenum (MFLU 21–0185). (a,b) Colony on decaying leaves; (c,d) co-
nidiophores, conidiogenous cells, and conidia; (e) conidiophores and conidiogenous cells; (f,g) co-
nidiogenous cells and conidia; (h–m) conidia; (n,o) colony cultures on PDA (observe and reverse).
Scale bars: (a) = 500 μm, (b) = 200 μm, (c–e) = 80 μm, (f–m) = 30 μm.

J. Fungi 2022, 8, 206 18 of 32
3.2.4. Neohelicosporium Y.Z. Lu, J.C. Kang, and K.D. Hyde, Mycological Progress 17: 637
(2017)
Index Fungorum: IF 822045
Type species: Neohelicosporium parvisporum Y.Z. Lu, J.C. Kang, and K.D. Hyde
Neohelicosporium was introduced by Lu et al. [10], with five new species. The taxo-
nomic revision of the genus was recently provided by Lu et al. [3]; eight Helicosporium,
two Helicoma, and one Tubeufia species were transferred to Neohelicosporium based on both
phylogeny and morphology. The genus is characterized by superficial, ellipsoidal to sub-
globose, ostiolate ascomata, bitunicate, cylindrical, pedicellate asci and fusiform, straight
or slightly curved, multi-septate, guttulate, hyaline, smooth-walled ascospores; mac-
ronematous, mononematous, branched or unbranched, septate, pale brown to brown co-
nidiophores, holoblastic, mono- to polyblastic, integrated, sympodial, intercalary or ter-
minal conidiogenous cells with denticles and solitary, acrogenous and/or acropleuroge-
nous, helicoid, multi-septate, guttulate, hyaline to pale brown conidia. Species of the ge-
nus are saprobic on decaying woody substrates from both aquatic and terrestrial habitats
[48]. In this study, the new species Neohelicosporium bambusicola is introduced based on
both phylogenetic analysis and morphological characters.
Neohelicosporium bambusicola X.G. Tian and Tibpromma, sp. nov. (Figure 5).
Index Fungorum number, IF 555045; Facesoffungi number, FoF 10571
Etymology: Referring to the host plant bamboo, on which the fungus was collected.
Saprobic on terrestrial dead culms of bamboo. Sexual morph Undetermined. Asexual
morph Hyphomycetous, helicosporous. Colonies on the substratum are superficial, effuse,
and white. Mycelium is composed of partly immersed, hyaline to brown, septate, branched
hyphae with glistening conidia. Conidiophores 21–76 × 3–5 μm (x = 48.5 × 4 μm, n = 10) are
macronematous, mononematous, cylindrical, unbranched or branched, septate, subhya-
line to brown, and smooth-walled. Conidiogenous cells 8.5–16 × 3–4.5 μm (x = 12 × 4 μm, n
= 15) are holoblastic, mono to ployblastic, integrated, sympodial, terminal or intercalary,
cylindrical, truncate at apex after conidial secession, pale brown, smooth-walled. Conidia
are solitary, acropleurogenous, helicoid, multi-septate, guttulate, hyaline when young
and become brown when mature, smooth-walled, and do not become loose in water, with
a 24–30 μm (x = 27 μm, n = 20) diam. and a conidial filament 3–4.5 μm (x = 3.9 μm, n = 20)
wide, 100.5–128 μm (x = 114 μm, n = 20) long, and coiled 2–2¾ times.
Culture characteristics: conidia germinated on PDA within 12 h; colonies on PDA
reach 20 mm in 2 weeks at 28 °C, and are superficial, effuse, and brown; mycelium is
composed of partly immersed, hyaline to brown, septate, branched, smooth hyphae.
Material examined: Thailand, Chiang Mai Province, on dead culms of bamboo, 16
December 2020, X. G. Tian, U4–10 (MFLU 21–0189 holotype), ex-type culture, MFLUCC
21–0156.
Notes: In the phylogenetic analyses, the new isolate Neohelicosporium bambusicola
(MFLUCC 21–0156) formed a distinct lineage sister to N. ellipsoideum (MFLUCC 16–0229)
and N. acrogenisporum (MFLUCC 17–2009). Neohelicosporium bambusicola resembles N. el-
lipsoideum and N. acrogenisporum in having macronematous, mononematous, unbranched
or branched, septate conidiophores, holoblastic, mono- to ployblastic conidiogenous cells,
and helicoid, septate conidia. However, Neohelicosporium bambusicola is distinct from N.
ellipsoideum and N. acrogenisporum as it has shorter and narrower conidiophores (21–76 ×
3–5 vs. 50–230 × 5–6 vs. 45–150 × 6–7 μm), smaller conidiogenous cells (8.5–16 × 3–4.5 vs.
15–25 × 5–6 vs. 12–15 × 4–6 μm), and narrower conidia (3–4.5 vs. 5–6 vs. 4.5–7.5 μm) [3].
Pairwise nucleotide comparisons revealed that the new isolate Neohelicosporium bam-
busicola is different from N. ellipsoideum (MFLUCC 16–0229) in 64/544 bp (11.76%) of the
ITS, 9/814 (1.1%) of the LSU, 20/1045 bp (1.91%) of RPB2, and 15/894 bp (1.68%) of TEF1-
α, while Neohelicosporium bambusicola is different from N. acrogenisporum (MFLUCC 17–
2009) in 2/370 bp (0.54%) of the ITS, 6/831 (0.72%) of the LSU, 27/1045 bp (2.58%) of RPB2,

J. Fungi 2022, 8, 206 19 of 32
and 16/894 bp (1.79%) of TEF1-α. Both phylogenetic analyses and morphological charac-
teristics support this species as a distinct new species.
Figure 5. Neohelicosporium bambusicola (MFLU 21–0189, holotype). (a,b) Colony on culms of bamboo;
(c,e–g,i) conidiogenous cells and conidia; (d) conidiophores; (h,k,l) conidia; (j) germinated conid-
ium; (m,n) colony cultures on PDA (observe and reverse). Scale bars: (a) = 500 μm, (b) = 200 μm,
(e,f,k,l) = 20 μm, (c,d,g–j) = 10 μm.

J. Fungi 2022, 8, 206 20 of 32
3.2.5. Parahelicomyces Goh, in Hsieh, and Goh and Kuo, Mycological Progress 20 (2): 182
(2021)
=Pseudohelicomyces Y.Z. Lu, J.K. Liu, and K.D. Hyde
Index Fungorum, IF 554886; Facesoffungi number, FoF 04745
Type species: Parahelicomyces talbotii (Goos) S.Y. Hsieh, Goh, and C.H. Kuo
Parahelicomyces is a well-studied genus, introduced as Pseudohelicomyces by Lu et al.
[3] with Pseudohelicomyces talbotii as the type species [3,49]. Pseudohelicomyces was renamed
Parahelicomyce by Hsieh et al. [50] because Parahelicomyces was a homonym and illegiti-
mate. Currently, seven species are accepted in the genus, and all the species have sequence
data available in the GenBank database. The genus is characterized by superficial, subglo-
bose, ellipsoidal-ovate, coriaceous, ostiolate ascomata, bitunicate, cylindrical, apically
thickened and rounded asci, and fusiform, multi-septate, hyaline, smooth-walled asco-
spores [3], as well as macronematous, mononematous, hyaline to brown, branched, sep-
tate conidiophores, holoblastic, mono- to polyblastic, integrated, intercalary or terminal,
determinate or sympodial conidiogenous cells with denticles and pleurogenous or
acropleurogenous, helicoid, multi-septate, hyaline to pale brown conidia. Species of the
genus Parahelicomyces are found from both terrestrial and freshwater habitats in China,
Japan, Mexico, South Africa, and Thailand [3,51]. In this study, we introduced a new Par-
ahelicomyces species from Thailand.
Parahelicomyces chiangmaiensis X.G. Tian and Tibpromma, sp. nov. (Figure 6).
Index Fungorum, IF 555060; Facesoffungi number, FoF 10570
Etymology: Referring to Chiangrai Province, Thailand, where the fungus was col-
lected.
Saprobic on a terrestrial woody substrate. Sexual morph Undetermined. Asexual
morph Hyphomycetous, helicosporous. Colonies on the substratum are superficial, effuse,
gregarious, and white. Mycelium is composed of partly superficial, hyaline to pale brown,
branched hyphae, with masses of crowded, glistening conidia. Conidiophores 85–180 × 2.9–
3.7 μm (x = 132 × 3 μm, n =10) are macronematous, mononematous, cylindrical, pale brown
to brown, paler towards the apex, straight or flexuous, branched, septate, and smooth-
walled. Conidiogenous cells 6–10 × 2.5–3.5 μm (x = 8 × 3 μm, n =20) are holoblastic, mono-
to polyblastic, integrated, sympodial, terminal or intercalary, cylindrical, with denticles,
hyaline to pale brown, and smooth-walled. Conidia have a 21–33.5 μm (x = 27 μm, n = 20)
diam. and a conidial filament 2–3 μm (x = 2.5 μm, n = 20) wide, 73–130 μm (x = 101.5 μm,
n = 20) long, and coiled 1¼–3 times, and are acropleurogenous, solitary, multi-septate,
helicoid, rounded at the tip, hyaline to pale brown, guttulate, tightly to loosely coiled in
water, and smooth-walled.
Culture characteristics: conidia germinated on PDA within 12 h; colonies adpressed
reaching 30 mm in 2 weeks at 28 °C, amd were circular, brown to dark brown, reverse
brown, and slow-growing; mycelium was superficial and partially immersed, branched,
septate, hyaline to pale brown, and smooth.
Material examined: Thailand, Chiang Mai Province, on the dead terrestrial woody
substrate, 16 December 2020, X. G. Tian, U4–8 (MFLU 21–0188 holotype), ex-type culture,
MFLUCC 21–0159.
Notes: Phylogenetic analyses of combined LSU, ITS, RPB2, and TEF1-α sequence data
showed that our new isolate Parahelicomyces chiangmaiensis (MFLUCC 21–0159) formed an
independent lineage within the genus with strong support (97% ML). Parahelicomyces
chiangmaiensis is phylogenetically closely related to P. talbotii (MFLUCC 17–2021), how-
ever, P. chiangmaiensis can be distinguished from P. talbotii by the size (21–33.5 vs. 7–16
μm diam.) of conidia and the size (6–10 vs. 7–16 μm long) of conidiogenous cells. Paraheli-
comyces chiangmaiensis is morphologically closely related to P. indicus, however, P.
chiangmaiensis can be distinguished from P. indicus by the colour (pale brown to brown vs.
dark to yellowish-brown) and size (85–180 × 2.9–3.7 vs. 47–145× 3–7.5 μm) of the

J. Fungi 2022, 8, 206 21 of 32
conidiophores [52]. Both phylogenetic analyses and morphological characteristics support
Parahelicomyces chiangmaiensis as a distinct new species.
Figure 6. Parahelicomyces chiangmaiensis (MFLU 21–0188, holotype). (a–b) Colony on decaying wood;
(c–g) conidiophores, conidiogenous cells, and conidia; (h–m) conidia; (n) germinated conidium; (o)
colony cultures on PDA (observe and reverse). Scale bars: (a) = 500 μm, (b) = 200 μm, (e,f,k–m) = 20
μm, (c,d,g–j,n) = 10 μm.
3.2.6. Tubeufia Penz. and Sacc., Malpighia 11(11–12): 517 (1898)
Index Fungorum: IF 5635
Type species: Tubeufia javanica Penz. and Sacc., Malpighia 11(11–12): 517 (1898)

J. Fungi 2022, 8, 206 22 of 32
Tubeufia, the type genus of Tubeufiaceae, was established by Penzig and Saccardo
[53]. Currently, 88 records are listed in the Index Fungorum (2021); however, most of the
species are lacking sequence data in the GenBank. While morphologies of Tubeufia species
are quite similar, using morphology alone presents difficulties for identification; thus, se-
quence data are required to resolve taxonomic confusions. The last treatment of Tubeufia
was provided by Lu et al. [3], and they introduced seventeen new species and six new
combinations in the genus, accepting fifty species in the genus based on both phylogenic
analysis and morphological characters. In this paper, we introduced two novel species,
one new record species, and a new isolate of known species in Tubeufia.
Tubeufia cocois X.G. Tian and Tibpromma, sp. nov. (Figure 7).
Index Fungorum number, IF 555070; Facesoffungi number, FoF 10576
Etymology: Referring to the host plant Cocos nucifera, on which the fungus was col-
lected.
Saprobic on the decaying leaves of Cocos nucifera. Sexual morph Undetermined. Asex-
ual morph Hyphomycetous, helicosporous. Colonies on the substratum are superficial, ef-
fuse, gregarious, and white to pale brown. Mycelium is partly immersed, partly superficial,
hyaline to brown, septate, branched, and with glistening conidia. Conidiophores 38–123 ×
4.5–6 μm (x = 80.5 × 5.5 μm, n = 20) are macronematous, mononematous, straight or
slightly flexuous, cylindrical, branched, septate, orange brown to dark brown, paler to-
wards the apex, and smooth-walled. Conidiogenous cells 8–17.5 × 4–5.5 μm (x = 13 × 5 μm,
n =25) are holoblastic, mono- to polyblastic, integrated, sympodial, terminal or intercalary,
irregular cylindrical, hyaline to pale brown, and smooth-walled, with most of them being
denticulate protrusions. Conidia are solitary, acropleurogenous, helicoid, rounded at tip,
with a 26–32.5 μm (x = 29 μm, n = 20) diam. and a conidial filament 3.5–5 μm (x = 4 μm, n
= 20) wide, 116–136 μm (x = 126 μm, n = 20) long, and coiled 2¼–2¾ times, and they do
not become loose in water, are indistinctly multi-septate, guttulate, hyaline when young,
pale brown to brown at maturity, and smooth-walled.
Culture characteristics: conidia germinated on PDA within 12 h; colonies grow on
PDA, reach 30 mm in 2 weeks at 28 °C, are irregular, with a flat surface, edge undulate,
and brown to dark brown in PDA medium; mycelium are superficial and partially im-
mersed, branched, septate, hyaline to brown, and smooth.
Material examined: Thailand, Chiang Rai Province, on decaying leaves of Cocos nu-
cifera, 16 January 2021, X. G. Tian, C6–15 (MFLU 21–0192, holotype), ex-type culture,
MFLUCC 22–0001; ibid, C6–8 (MFLU 21–0186, paratype), ex-paratype, MFLUCC 22–0002;
ibid, C6–20 (MFLU 21–0187, paratype), ex-paratype, MFLUCC 22–0003.
Notes: Tubeufia cocois is introduced as a distinct new species from Cocos nucifera in
Thailand. In the phylogenetic analyses, three newly obtained strains of T. cocois (MFLUCC
22–0001, MFLUCC 22–0002, and MFLUCC 22–0003) clustered together and were sister to
three T. aquatica strains with strong statistical support values (100% ML/1.00 PP, Figure
1). Morphologically, T. cocois can be easily distinguished from T. aquatica by the shape and
size of the conidiophores, conidiogenous cells, and conidia. Tubeufia cocois has branched
or unbranched, multi-septate, and longer conidiophores (38–123 vs. 18–40 μm), while the
conidiophores of T. aquatica are unbranched, 0–1-septate, and shorter than those of T. co-
cois. The conidiogenous cells of T. cocois are terminal or intercalary and conidia are
acropleurogenous, whereases T. aquatica has terminal conidiogenous cells and conidia are
acrogenous [54]. Based on pairwise nucleotide comparisons, the new strain (MFLUCC 22-
0001) is different from T. aquatica (MFLUCC 16–1249) in 16/413 bp (3.87%) of the ITS, 2/845
(0.24%) of the LSU, 23/919 bp (2.5%) of RPB2, and 10/617 bp (1.62%) of TEF1-α.

J. Fungi 2022, 8, 206 23 of 32
Figure 7. Tubeufia cocois (MFLU 21–0192, holotype). (a) Colony on decaying leaves of Cocos nucifera;
(b,c,e) conidiophores and conidiogenous cells; (d) conidia and conidiogenous cells; (f–l) conidia;
(m) germinated conidium; (n,o) colony cultures on PDA (observe and reverse). Scale bars: (b–m) =
20 μm.
Tubeufia laxispora Y.Z. Lu, Boonmee, and K.D. Hyde, Mycological Progress 16: 409 (2017)
(Figure 8).
Index Fungorum, IF 818987; Facesoffungi number, FoF 02694

J. Fungi 2022, 8, 206 24 of 32
Saprobic on the decaying leaves of Cocos nucifera. Sexual morph Undetermined. Asex-
ual morph Hyphomycetous, helicosporous. Colonies on the substratum are superficial, ef-
fuse, gregarious, and range from white to brown. Mycelium is partly immersed, partly
superficial, pale brown, septate, sparsely branched hyphae, with masses of crowded co-
nidia. Conidiophores 26–53 × 3.5–5 μm (x = 39.5 × 4 μm, n =15) are hyaline to brown, mac-
ronematous, erect, short, and smooth-walled. Conidiogenous cells 8–16 × 3–4.5 μm (x = 12 ×
4 μm, n =20) are monoblastic, holoblastic, integrated, and each have a single conidium.
Conidia are solitary, acropleurogenous, helicoid, and rounded at the tip, with a 17.5–29 μm
(x = 23 μm, n = 20) diam. and a conidial filament 2–3.5 μm (x = 2.5 μm, n = 20) wide, 63–
94.5 μm (x = 78.5 μm, n = 20) long, loosely coiled 1–2½ times in the water, indistinctly
multi-septate, hyaline, and smooth-walled.
Culture characteristics: conidia germinating on PDA within 12 h; colonies growing
on PDA, reaching 20 mm in 2 weeks at 28 °C—they are circular, with a flat surface, edge
undulate, and brown to dark brown in PDA medium; mycelium are superficial and par-
tially immersed, branched, septate, hyaline to brown, and smooth.
Material examined: Thailand, Chiang Rai Province, on decaying leaves of Cocos nu-
cifera, 9 March 2021, X. G. Tian, C7–10 (MFLU 21–0191), living culture, MFLUCC 21–0163.
Notes: In our phylogenetic analyses, the newly obtained isolate (MFLUCC 21–0163)
clustered with four strains of T. laxispora with high statistical supports (100% ML/1.00 PP,
Figure 1). Based on pairwise nucleotide comparisons, the new strain (MFLUCC 21–0163)
almost overlapped with the ex-type strain of T. laxispora (MFLUCC 16–0232), except TEF1-
α 1 bp out of 878 bp (<1%). Morphologically, our new isolate fits well with the description
of T. laxispora, except for the conidial size (17.5–40 μm diam., 63–94.5 μm long vs. 17.5–29
μm diam., 111–182 μm long) [38]. Hence, we identify our new isolate as T. laxispora.
Tubeufia laxispora was described by Lu et al. [38] on submerged wood in Thailand, while
our new isolate was collected on decaying leaves of Cocos nucifera in Thailand, and this is
the first report of Tubeufia laxispora associated with a coconut tree from Thailand.
Tubeufia cylindrothecia (Seaver) Ho¨hn Sber. Akad. Wiss. Wien, Math. -naturw. Kl., Abt.
1 128: 562 (1919) (Figure 9).
Index Fungorum, IF 340543; Facesoffungi number, FoF 02650
Saprobic on submerged decaying wood. Sexual morph See Seaver [55]. Asexual
morph Colonies on the substratum are superficial, effuse, gregarious, and white to pale
brown. Mycelium is partly immersed, partly superficial, hyaline to brown, septate, and
with masses of conidia. Conidiophores 57–95 × 4–7 μm (x = 76 × 5.5 μm, n =10) are pale
brown, macronematous, mononematous, septate, cylindrical, unbranched, erect, and
smooth-walled. Conidiogenous cells 6.5–16 × 3.5–5 μm (x = 11 × 4 μm, n =15) are holoblastic,
monoblastic, integrated, smooth, terminal or intercalary, and cylindrical. Conidia are
acropleurogenous, 40.5–82 μm (x = 61.5 μm, n = 20) in diam. and with a conidial filament
that is 3.5–8 μm (x = 5.5 μm, n = 20) wide, 220–321 μm (x = 270.5 μm, n = 20) long, helicoid,
with conidial loosely coiled 1–3½ times in the water, hyaline to brown, indistinctly multi-
septate, guttulate, and smooth.
Culture characteristics: conidia germinating on PDA within 12 h; colonies growing
on PDA, reaching 25 mm in 2 weeks at 28 °C, irregular, with a flat surface, edge undulate,
and brown to dark brown in PDA medium; mycelium are partially immersed, branched,
septate, hyaline to brown, and smooth.
Material examined: Thailand, Chiang Rai Province, on decaying submerged wood,
11 November 2020, X. G. Tian, W1–10 (MFLU 21–0190), living culture, MFLUCC 21–0160.

J. Fungi 2022, 8, 206 25 of 32
Figure 8. Tubeufia laxispora (MFLU 21–0191). (a,b) Colony on decaying leaves of Cocos nucifera; (c–f)
conidiogenous cells and conidia; (g–m) conidia; (n) germinated conidium; (o,p) colony cultures on
PDA (observe and reverse). Scale bars: (c–n) = 20 μm.
Notes: Tubeufia cylindrothecia was originally introduced with both sexual and asexual
morphs that link to Helicomyces roseus based on morphological studies [6,56], while phy-
logenetic analyses showed that Tubeufia cylindrotheci and Helicomyces roseus can be recog-
nized as two different species [1,8,54,57]. Luo et al. [54] first reported its asexual morph as
collected from a freshwater habitat in China. Our phylogenetic results show that the
newly obtained isolate (MFLUCC 21–0160) clustered with six strains of T. cylindrothecia
with high bootstrap support (98% ML/1.00 PP). Morphologically, our new isolate is almost
identical to T. cylindrothecia, except for the conidiogenous cells of the new isolate
(MFLUCC 21–0160) that are terminal or intercalary, while the conidiogenous cells are ter-
minal in T. cylindrothecia (MFLU 16–2547). Thus, based on morphological and molecular
data, we identify the new isolate as Tubeufia cylindrothecia.

J. Fungi 2022, 8, 206 26 of 32
Figure 9. Tubeufia cylindrothecia (MFLU 21–0190). (a,b) Colony on decaying wood; (c,e,f) conidio-
phores with attached conidia; (d,g) conidiophores and conidiogenous cells; (h–k,m,n) conidia; (l)
germinated conidium; (o,p) colony cultures on PDA (observe and reverse). Scale bars: (a) = 1 000
μm, (b) = 200 μm, (l) = 50 μm, (c–f,h) = 30 μm, (i–k,m,n) = 20 μm, (g) = 10 μm.
Tubeufia longihelicospora Boonmee, Promputtha, and K.D. Hyde, in Boonmee et al., Fun-
gal Diversity 111: 133 (2021) (Figure 10).
Index Fungorum number, IF 558543; Facesoffungi number, FoF 09195

J. Fungi 2022, 8, 206 27 of 32
Saprobic on submerged decaying wood in a freshwater stream. Sexual morph Unde-
termined. Asexual morph Hyphomycetous, helicosporous. Colonies on the substratum are
superficial, effuse, gregarious, and white. Mycelium is composed of partly immersed, hy-
aline to pale brown, septate, branched hyphae with masses of glistening, crowded conidia.
Stalked sclerotia are often present and are medium brown, spherical, and muriform. Co-
nidiophores (10.5–)13.5–35.5(–55.5) × 3.5–5(–6) μm (x = 24.5 × 4 μm, n = 15) are macronema-
tous, mononematous, arise as lateral branches from creeping hyphae, cylindrical,
branched, 0–4-septate, hyaline to pale brown, and smooth-walled. Conidiogenous cells (7–
)9–19(–24.5) × 3.5–4.5 μm (x = 14 × 4 μm, n = 15) are holoblastic, monoblastic, integrated,
terminal or intercalary, cylindrical, truncate at the apex after conidial secession, hyaline
to pale brown, and smooth-walled. Conidia are solitary, acrogenous, holoblastic, helicoid,
rounded at the tip, hyaline to pale brown, (40–)55.5–86.5(–100.5) μm (x = 71 μm, n = 15) in
diam. and with a conidial filament 6.5–8.5 μm (x = 7.5 μm, n = 15) wide in the broadest
part and tapering towards the ends, (99.5–)240.5–355.5 μm (x = 298 μm, n = 15) long,
loosely coiled 1–2 times, multi-septate, tightly to loosely coiled in the water, constricted at
the septa, guttulate, hyaline to pale brown, rough-walled, and bearing conidiola. Conidi-
ola are globose, unicellular, and rough-walled.
Culture characteristics: conidia germinated on PDA within 12 h; colonies growing on
PDA reach 20 mm in 2 weeks at 28 °C, are irregular, with a flat surface, edge undulate,
and brown to dark brown in PDA medium; mycelium are superficial and partially im-
mersed, branched, septate, hyaline to brown, and smooth.
Material examined: Thailand, Chiang Rai Province, Mueang, Ban Du, on decaying
submerged wood, 15 August 2020, R. J. Xu, MD77 (MFLU 21–0182), living culture,
MFLUCC 21–0151; China, Yunnan Province, Xishuangbanna, on decaying submerged
wood, 13 September 2021, X. G. Tian, WB12 (HKAS 122173), living culture, KUMCC 21–
0814; ibid NWBB9 (HKAS 122169), living culture, KUMCC 21–0815.
Notes: In the phylogenetic analyses, our three new strains (MFLUCC 21–0151,
KUMCC 21–0478, and KUMCC 21–0479) are clustered together within the same clade as
T. longihelicospora (MFLUCC 16–0753). Based on pairwise nucleotide comparisons, our
three new strains almost overlap with the ex-type strain of Tubeufia longihelicospora
(MFLUCC 16–0753). Morphologically, our new isolate (MFLU 21–0182) is almost identical
to Tubeufia longihelicospora, except for the size of the conidia (55.5–86.5 vs. 36–52 μm diam.).
Therefore, we identify the three new isolates as Tubeufia longihelicospora based on morpho-
logical and phylogenetic data. Tubeufia longihelicospora was introduced by Boonmee et al.
[58] on a submerged decaying wood in a small freshwater stream in Thailand. Our isolate
Tubeufia longihelicospora (MFLUCC 21–0151) was also collected in Thailand, while the
other two isolates of Tubeufia longihelicospora (KUMCC 21–0478 and KUMCC 21–0479)
were collected in China, which is a new geographical record.

J. Fungi 2022, 8, 206 28 of 32
Figure 10. Tubeufia longihelicospora (MFLU 21–0182). (a–b) Colony on decaying wood; (c–f) conidio-
phores, conidiogenous cells, and conidia; (g–h) stalked sclerotia; (i–m) conidia; (n) germinated co-
nidium; (o) colony cultures on PDA (observe and reverse). Scale bars: (n) = 40 μm, (c–m) = 20 μm.
4. Discussion
Tubeufiaceae is an interesting family with diverse morphologies, habitats, and a
worldwide distribution [1,3,12,25]. The asexual morph of the Tubeufiaceae species is re-
ported as helicosporous, chlamydosporous, and phragmosporous conidia, of which heli-
cosporous conidia are the most common morphology in Tubeufiaceae. However, species
with helicosporous conidia are not only found in Tubeufiaceae; for example,

J. Fungi 2022, 8, 206 29 of 32
Helicoascotaiwania also has helicosporous conidia but it is phylogenetically distinct from
Tubeufiaceae. Helicoascotaiwania is placed in Pleurotheciaceae and Sordariomycetes [59],
while Tubeufiaceae is placed in Pleosporales. The interesting finding is that the asexual
morphs of some genera were reported with two different morphologies. For example,
Tubeufia and Berkleasmium produce both dictyosporous and helicosporous conidia, while
Helicoma produces both helicosporous and phragmosporous conidia, confirmed by phy-
logenetic analyses [3].
The morphology of helicosporous hyphomycetes is quite similar; thus, the phyloge-
netic analyses are efficient to identify the helicosporous hyphomycetes at the species level
[60–62]. With the availability of molecular data, some species were revised and transferred
to other genera based on phylogenetic analyses. For example, Helicomyces roseus (BCC
3381) and Helicoma perelegans (ATCC 22621) were transferred to Tubeufia [3]. However,
many helicosporous species lack sequence data in the GenBank, and the taxonomy of hel-
icosporous hyphomycetes needs revisions based on phylogenetic analyses. For example,
species in the genera of Tubeufia, Helicoma, and Helicomyces were introduced based on mor-
phological characteristics, but many of them lack sequence data in the GenBank; thus,
further study into herbarium specimens is necessary to resolve taxonomic problems in the
three genera.
In this study, nine Tubeufiaceae species were collected from terrestrial and freshwa-
ter habitats in Thailand, of which three were introduced as new species, while six were
identified as existing species based on phylogenetic analyses and morphological charac-
teristics. The nine species were placed in Dematiohelicomyces, Helicoma, Helicotruncatum,
Neohelicosporium, Parahelicomyces, and Tubeufia, respectively, of which the genera Dematio-
helicomyces, Helicotruncatum, Neohelicosporium, and Parahelicomyces are well studied, and
all species in these genera have sequence data available in the GenBank. Helicoma and
Tubeufia were recently revised by Lu et al. [3]. In our phylogenetic analyses, Helicoma and
Tubeufia formed well-supported and monophyletics clades within the family. The mor-
phologies of Tubeufia and Helicoma are quite similar; thus, morphology alone is not enough
to identify species in Tubeufia and Helicoma, and phylogenetic analyses are necessary.
However, earlier studies identified the two genera only based on morphological charac-
teristics, and sequence data of many species are not available in the GenBank, so, it is
entirely possible that some species were incorrectly identified; accordingly, fresh collec-
tions and molecular data are required to clarify their taxonomic status. Even though De-
matiohelicomyces helicosporus, Helicotruncatum palmigenum, Helicoma guttulatum, Neohelico-
sporium bambusicola, Tubeufia cylindrothecia, T. laxispora, and T. longihelicospora are known
species and were collected again, some species are known as new hosts and new geo-
graphical records. In addition, it is also better to provide the full descriptions and color
plates of the micro-characteristics of new isolates to understand some fine morphological
differences.
Author Contributions: Conceptualization, X.T.; formal analysis, X.T.; funding acquisition, S.C.K.
and S.T.; methodology, D.B. and R.X.; resources, S.C.K. and S.T.; supervision, S.C.K.; writing—orig-
inal draft, X.T.; writing—review and editing, X.T., J.X., A.M., S.C.K., S.T., N.S. and Y.L. All authors
have read and agreed to the published version of the manuscript.
Funding: The APC was funded by editorial board member Dr. Samantha C. Karunarathna’s free
APC facility in 2021. Xingguo Tian thanks Prof. Kevin D. Hyde for reviewing and editing the man-
uscript. Saowaluck Tibpromma thanks the International Postdoctoral Exchange Fellowship Pro-
gram (number Y9180822S1), the CAS President’s International Fellowship Initiative (PIFI) (number
2020PC0009), the China Postdoctoral Science Foundation, and the Yunnan Human Resources and
Social Security Department Foundation for funding her postdoctoral research. Samantha C. Karuna-
rathna thanks the CAS President’s International Fellowship Initiative (PIFI) young staff under the
grant number: 2020FYC0002, and the National Science Foundation of China (NSFC) project
code31851110759 for funding. Yong-Zhong Lu would like to thank the National Natural Science
Foundation of China (NSFC 31900020) and the Science and Technology Foundation of Guizhou
Province ([2020]1Y058). Ausana Mapook would like to thank the MFU foundation grant and Mae

J. Fungi 2022, 8, 206 30 of 32
Fah Luang University Fund (Grant No. 651A16029), entitled “Taxonomy, phylogeny, risk assess-
ment, and potential impact of fungi on Siam weed in northern Thailand”. Austin Smith at World
Agroforestry (ICRAF), Kunming Institute of Botany, China, is thanked for English editing.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.
References
1. Boonmee, S.; Rossman, A.Y.; Liu, J.K.; Li, W.J.; Dai, D.Q.; Bhat, J.D.; Jones, E.B.G.; McKenzie, E.H.C.; Xu, J.C.; Hyde, K.D.
Tubeufiales, ord. nov., integrating sexual and asexual generic names. Fungal Divers 2014, 68, 239–298.
2. Liu, J.K.; Hyde, K.D.; Jeewon, R.; Phillips, A.J.; Maharachchikumbura, S.S.; Ryberg, M.; Liu, Z.Y.; Zhao, Q. Ranking higher taxa
using divergence times: A case study in Dothideomycetes. Fungal Divers 2017, 84, 75–99.
3. Lu, Y.Z.; Liu, J.K.; Hyde, K.D.; Jeewon, R.; Kang, J.C.; Fan, C.; Boonmee, S.; Bhat, D.J.; Luo, Z.L.; Lin, C.G.; et al. A taxonomic
reassessment of Tubeufiales based on multi-locus phylogeny and morphology. Fungal Divers 2018, 92, 131–344.
4. Hongsanan, S.; Hyde, K.D.; Phookamsak, R.; Wanasinghe, D.N.; McKenzie, E.H.; Sarma, V.V.; Lücking, R.; Boonmee, S.; Bhat,
J.D.; Liu, N.G.; et al. Refined families of Dothideomycetes: Orders and families incertae sedis in Dothideomycetes. Fungal Divers
2020, 105, 17–318.
5. Barr, M.E. A classification of Loculoascomycetes. Mycologia 1979, 71, 935–957.
6. Barr, M.E. On the family Tubeufiaceae (Pleosporales). Mycotaxon 1980, 12, 137–167.
7. Rossman, A.Y. The Tubeufiaceae and similar Loculoascomycetes; CAB International Mycological Institute: London, UK, 1987.
8. Boonmee, S.; Zhang, Y.; Chomnunti, P.; Chukeatirote, E.; Tsui, C.K.; Bahkali, A.H.; Hyde, K.D. Revision of lignicolous
Tubeufiaceae based on morphological reexamination and phylogenetic analysis. Fungal Divers 2011, 51, 63–102.
9. Brahmanage, R.S.; Lu, Y.Z.; Bhat, D.J.; Wanasinghe, D.N.; Yan, J.Y.; Hyde, K.D.; Boonmee, S. Phylogenetic investigations on
freshwater fungi in Tubeufiaceae (Tubeufiales) reveals the new genus Dictyospora and new species Chlamydotubeufia aquatica
and Helicosporium flavum. Mycosphere 2017, 8, 917–933.
10. Lu, Y.Z.; Boonmee, S.; Liu, J.K.; Hyde, K.D.; McKenzie, E.H.; Eungwanichayapant, P.D.; Kang, J.C. Multi-gene phylogenetic
analyses reveals Neohelicosporium gen. nov. and five new species of helicosporous hyphomycetes from aquatic habitats. Mycol.
Prog. 2018, 17, 631–646.
11. Rajeshkumar, K.C.; Sharma, R. Tamhinispora, a new genus belongs to family Tubeufiaceae from the western Ghats, India based
on morphology and phylogenetic analysis. Mycosphere 2013, 4, 165–174.
12. Dong, W.; Wang, B.; Hyde, K.D.; McKenzie, E.H.C.; Raja, H.A.; Tanaka, K.; Abdel-Wahab, M.A.; Abdel-Aziz, F.A.; Doilom, M.;
Phookamsak, R.; et al. Freshwater Dothideomycetes. Fungal Divers 2020, 105, 319–575.
13. Sivanesan, A. The bitunicate ascomycetes and their anamorphs. J. Cramer. Vaduz. 1984, pp. 701.
14. Crane, J.L.; Shearer, C.J.; Barr, M.E. A revision of Boerlagiomyces with notes and a key to the saprobic genera of Tubeufiaceae.
Can. J. Bot. 1998, 76, 602–612.
15. Kirk, P.M.; Cannon, P.F.; David, J.C.; Stalpers, J.A. Ainsworth & Bisby’s Dictionary of the Fungi, 9th ed.; CABI International: Wall-
ingford, UK, 2001; pp. 1–655.
16. Eriksson, O.E.; Winka, K. Families and higher taxa of Ascomycota. Myconet 1998, 1, 17–24.
17. Eriksson, O.E. Outline of Ascomycota—1999. Myconet 1999, 3, 1–88.
18. Eriksson, O.E. SSU rDNA sequences from Ascomycota. Myconet 2001, 6, 27–76.
19. Eriksson, O.E. Outline of Ascomycota—2005. Myconet 2005, 11, 1–113.
20. Lumbsch, H.T.; Huhndorf, S.M. Myconet volume 14. Part One. Outline of Ascomycota—2009. Part Two. Notes on Ascomycete
systematics. Nos. 4751–5113. Fieldiana Life Earth Sci. 2010, 2010, 1–64.
21. Kodsueb, R.; Jeewon, R.; Vijaykrishna, D.; McKenzie, E.H.C.; Lumyong, P.; Lumyong, S.; Hyde, K.D. Systematic revision of
Tubeufiaceae based on morphological and molecular data. Fungal Divers 2006, 21, 105–130.
22. Rossman, A.Y. The Tubeufiaceae and similar Loculoascomycetes. Mycol. Pap. 1987, 157, 1–71.
23. Chaiwan, N.; Lu, Y.Z.; Tibpromma, S.; Bhat, D.J.; Hyde, K.D.; Boonmee, S. Neotubeufia gen. nov. and Tubeufia guangxiensis sp.
nov. (Tubeufiaceae) from freshwater habitats. Mycosphere 2017, 8, 1443–1456.
24. Hanada, T.; Sato, T.; Arioka, M.; Uramoto, M.; Yamasaki, M. Purification and characterization of a 15 kDa protein (p15) pro-
duced by Helicosporium that exhibits distinct effects on neurite outgrowth from cortical neurons and PC12 cells. Biochem. Bioph.
Rre. Co. 1996, 228, 209–215.
25. Hu, H.; Guo, H.; Li, E.; Liu, X.; Zhou, Y.; Che, Y. Decaspirones F–I, bioactive secondary metabolites from the saprophytic fungus
Helicoma viridis. J. Nat. Prod. 2006, 69, 1672–1675.
26. Itazaki, H.; Nagashima, K.; Sugita, K.; Yoshida, H.; Kawamura, Y.; Yasuda, Y.; Matsumoto, K.; Ishii, K.; Uotani, N.; Nakai, H.
Solation and structural elucidation of new cyclotetrapeptides, trapoxins A and B, having detransformation activities as anti-
tumor agents. J. Antibiot. 1990, 43, 1524–1532.

J. Fungi 2022, 8, 206 31 of 32
27. Tibpromma, S.; Hyde, K.D.; Jeewon, R.; Maharachchikumbura, S.S.; Liu, J.K.; Bhat, D.J.; Jones, E.G.; McKenzie, E.H.; Camporesi,
E.; Bulgakov, T.S.; et al. Fungal diversity notes 491–602: Taxonomic and phylogenetic contributions to fungal taxa. Fungal Divers
2017, 83, 1–261.
28. Tian, X.G.; Karunarathna, S.C.; Mapook, A.; Xu, J.C.; Bao, D.F.; Promputtha, I.; Tibpromma, S. Koorchaloma oryzae sp. nov.
(Stachybotryaceae, Sordariomycetes), from Oryza sativa (Poaceae) in northern Thailand. Phytotaxa 2021, 524, 283–292.
29. Tian, X.G.; Karunarathna, S.C.; Mapook, A.; Promputtha, I.; Xu, J.C.; Bao, D.F.; Tibpromma, S. One new species and two new
host records of Apiospora from bamboo and maize in northern Thailand with thirteen new combinations. Life 2021, 11, 1071.
30. Senanayake, I.C.; Rathnayaka, A.R.; Marasinghe, D.S.; Calabon, M.S.; Gentekaki, E.; Lee, H.B.; Hurdeal, V.G.; Pem, D.; Dis-
sanayake, L.S.; Wijesinghe, S.N.; et al. Morphological approaches in studying fungi: Collection, examination, isolation, sporu-
lation and preservation. Mycosphere 2020, 11, 2678–2754.
31. Index Fungorum. 2021. Available online: http://www.indexfungorum.org/Names/Names.asp (accessed on 30 November 2021).
32. Jayasiri, S.C.; Hyde, K.D.; Ariyawansa, H.A.; Bhat, J.; Buyck, B.; Cai, L.; Dai, Y.C.; Abd-Elsalam, K.A.; Ertz, D.; Hidayat, I.; et al.
The faces of fungi database: Fungal names linked with morphology, phylogeny and human impacts. Fungal Divers 2015, 74, 3–
18.
33. White, T.J.; Bruns, T.; Lee, S.; Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.
In PCR Protocols: A Guide to Methods and Applications; Innis, G.M., Shinsky, D., White, T., Eds.; Academic: New York, USA, 1990;
pp. 315–322.
34. Vilgalys, R.; Hester, M. Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several
Cryptococcus species. J. Bacteriol. 1990, 172, 4238–4246.
35. Rehner, S.A.; Buckley, E. A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: Evidence for cryptic diversi-
fication and links to Cordyceps teleomorphs. Mycologia 2005, 97, 84–98.
36. Liu, Y.J.; Whelen, S.; Hall, B.D. Phylogenetic relationships among ascomycetes: Evidence from an RNA polymerse II subunit.
Mol. Biol. Evol. 1999, 16, 1799–1808.
37. Cai, L.; Jeewon, R.; Hyde, K.D. Phylogenetic evaluation and taxonomic revision of Schizothecium based on ribosomal DNA and
protein coding genes. Fungal Divers 2005, 19, 1–21.
38. Lu, Y.Z.; Boonmee, S.; Dai, D.Q.; Liu, J.K.; Hyde, K.D.; Bhat, D.J.; Ariyawansa, H.; Kang, J.C. Four new species of Tubeufia
(Tubeufiaceae, Tubeufiales) from Thailand. Mycol. Prog. 2017, 16, 403–417.
39. Dissanayake, A.J.; Bhunjun, C.S.; Maharachchikumbura, S.S.N.; Liu, J.K. Applied aspects of methods to infer phylogenetic rela-
tionships amongst fungi. Mycosphere 2020, 11, 2652–2676.
40. Katoh, K.; Rozewicki, J.; Yamada, K.D. MAFFT online service: Multiple sequence alignment, interactive sequence choice and
visualization. Brief. Bioinform. 2019, 20, 1160–1166.
41. Capella-Gutiérrez, S.; Silla-Martínez, J.M.; Gabaldón, T. TrimAl: A tool for automated alignment trimming in large-scale phy-
logenetic analyses. Bioinformatics 2009, 25, 1972–1973.
42. Stamatakis, A.; Hoover, P.; Rougemont, J. A rapid bootstrap algorithm for the RAxML web servers. Syst. Biol. 2008, 57, 758–771.
43. Miller, M.A.; Pfeiffer, W.; Schwartz, T. Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In Pro-
ceedings of the 2010 Gateway Computing Environments Workshop (GCE), New Orleans, LA, USA, 14 November 2010; pp. 1–
8.
44. Hyde, K.D.; Hongsanan, S.; Jeewon, R.; Bhat, D.J.; McKenzie, E.H.C.; Jones, E.B.G.; Phookamsak, R.; Ariyawansa, H.A.; Boon-
mee, S.; Zhao, Q.; et al. Fungal diversity notes 367–490: Taxonomic and phylogenetic contributions to fungal taxa. Fungal Divers
2016, 80, 1–270.
45. Corda, A.K.J. Incones fungorum hucusque cognitorum. J. G. Calve 1837, 1, 1–32.
46. Linder, D.H. A monograph of the helicosporous fungi imperfecti. Ann. Missouri Bot. Gard. 1929, 16, 227–388.
47. Goos, R.D. A review of the anamorph genus Helicoma. Mycologia 1986, 78, 744–761.
48. Li, W.J.; McKenzie, E.H.C.; Liu, J.K.; Bhat, D.J.; Dai, D.Q.; Camporesi, E.; Tian, Q.; Maharachchikumbura, S.S.N.; Luo, Z.L.;
Shang, Q.J.; et al. Taxonomy and phylogeny of hyaline-spored coelomycetes. Fungal Divers 2020, 100, 279–801.
49. Lu, Y.Z.; Liu, J.K.; Hyde, K.D. Proposal to conserve Pseudohelicomyces YZ Lu & al. (Tubeufiaceae) against Pseudohelicomyces
Garnica & E. Valenz. (Hymenogastraceae). Taxon 2020, 69, 615–616.
50. Hsieh, S.Y.; Goh, T.K.; Kuo, C.H. New species and records of Helicosporium sensu lato from Taiwan, with a reflection on current
generic circumscription. Mycol. Prog. 2021, 20, 169–190.
51. Rossman, A.Y. The genus Ophionectria (Euascomycetes, Hypocreales). Mycologia 1977, 69, 355–391.
52. Rao, P.R.; Rao, D. Some helicosporae from Hyderabad-II. Mycopathol. Mycol. Appl. 1964, 24, 27–34.
53. Penzig, O.A.J.; Saccardo, P.A. Diagnoses fungorum novorum in Insula Java collectorum. Series secunda. Malpighia 1897, 11,
491–530.
54. Luo, Z.L.; Bhat, D.J.; Jeewon, R.; Boonmee, S.; Bao, D.F.; Zhao, Y.C.; Chai, H.M.; Su, H.Y.; Su, X.J.; Hyde, K.D. Molecular phy-
logeny and morphological characterization of asexual fungi (Tubeufiaceae) from freshwater habitats in Yunnan, China. Crypto-
gam. Mycol. 2017, 38, 27–53.
55. Seaver, F.J. The Hypocreales of north America–I. Mycologia 1909, 1, 41–76.
56. Seaver, F.J.; Waterston, J.M. Contributions to the mycoflora of Bermuda–I. Mycologia 1940, 32, 388–407.
57. Zhao, G.Z.; Liu, X.Z.; Wu, W.P. Helicosporous hyphomycetes from China. Fungal Divers 2007, 26, 313–524.

J. Fungi 2022, 8, 206 32 of 32
58. Boonmee, S.; Wanasinghe, D.N.; Calabon, M.S.; Huanraluek, N.; Chandrasiri, S.K.; Jones, G.E.; Rossi, W.; Leonardi, M.; Singh,
S.K.; Rana, S.; et al. Fungal diversity notes 1387–1511: Taxonomic and phylogenetic contributions on genera and species of
fungal taxa. Fungal Divers 2021, 111, 1–335.
59. Dayarathne, M.C.; Maharachchikumbura, S.S.N.; Jones, E.B.G.; Dong, W.; Devadatha, B.; Yang, J.; Ekanayaka, A.H.; De Silva,
W.; Sarma, V.V.; Al-Sadi, A.M.; et al. Phylogenetic revision of Savoryellaceae and evidence for its ranking as a subclass. Front.
Microbiol. 2019, 10, 840.
60. Chethana, K.W.T.; Manawasinghe, I.S.; Hurdeal, V.G.; Bhunjun, C.S.; Appadoo, M.A.; Gentekaki, E.; Raspé, O.; Promputtha, I.;
Hyde, K.D. What are fungal species and how to delineate them. Fungal Divers 2021, 109, 1–25.
61. Pem, D.; Jeewon, R.; Chethana, K.W.T.; Hongsanan, S.; Doilom, M.; Suwannarach, N.; Hyde, K.D. Species concepts of Dothide-
omycetes: Classification, phylogenetic inconsistencies and taxonomic standardization. Fungal Divers 2021, 109, 283–319.
62. Hyde, K.D.; Jeewon, R.; Chen, Y.J.; Bhunju, C.S.; Calabon, M.S.; Jiang, H.B.; Lin, C.G.; Norphanphoun, C.; Sysouphanthong, P.;
Pem, D.; et al. The numbers of fungi: Is the descriptive curve flattening? Fungal Divers 2020, 103, 219–271.