Yuxiensis granularis gen. et sp. nov., a Novel Quellkörper-Bearing Fungal Taxon Added to Scortechiniaceae and Inclusion of Parasympodiellaceae in Coronophorales Based on Phylogenetic Evidence

Abstract

An undetermined saprobic fungal taxon from Yunnan (China) is revealed as a new genus in Scortechiniaceae (Coronophorales). The novel taxon, Yuxiensis, is characterized by immersed to erumpent, semi-globose ascomata, which are not surrounded by any tomentum or conspicuous subiculum, a subcylindrical quellkörper in the centrum, clavate asci with long pedicels and allantoid hyaline ascospores with granular contents. Maximum likelihood and Bayesian posterior probability analyses based on LSU, ITS, tef1 and rpb2 sequence data depict a close phylogenetic relationship of the new genus to Pseudocatenomycopsis, hence, confirming its placement in Scortechiniaceae. Parasympodiellaceae, thus far belonging to Parasympodiellales, is transferred to Coronophorales based on multi-gene phylogenetic evidence. Additionally, the incertae sedis monotypic genus Arthrocristula is treated as a synonym of Parasympodiella, with Arthrocristula hyphenata recombined as Parasympodiella hyphenata comb. nov., as the type strain of Arthrocristula hyphenata clusters inside the Parasympodiellaceae clade along with other Parasympodiella taxa.

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Article
Yuxiensis granularis gen. et sp. nov., a Novel Quellkörper-Bearing
Fungal Taxon Added to Scortechiniaceae and Inclusion of
Parasympodiellaceae in Coronophorales Based on
Phylogenetic Evidence
Digvijayini Bundhun 1,2,3, Dhanushka N. Wanasinghe 1,4, *, Sajeewa S. N. Maharachchikumbura 5,
Darbhe J. Bhat 6, Shi-Ke Huang 3, Saisamorn Lumyong 7,8,9, Peter E. Mortimer 1,4 and Kevin D. Hyde 2,3,7,10,*


Citation: Bundhun, D.; Wanasinghe,
D.N.; Maharachchikumbura, S.S.N.;
Bhat, D.J.; Huang, S.-K.; Lumyong, S.;
Mortimer, P.E.; Hyde, K.D. Yuxiensis
granularis gen. et sp. nov., a Novel
Quellkörper-Bearing Fungal Taxon
Added to Scortechiniaceae and
Inclusion of Parasympodiellaceae in
Coronophorales Based on Phylogenetic
Evidence. Life 2021,11, 1011. https://
doi.org/10.3390/life11101011
Academic Editor: Francois Lefort
Received: 30 August 2021
Accepted: 19 September 2021
Published: 25 September 2021
Publisher’s Note: MDPI stays neutral
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Copyright: © 2021 by the authors.
Licensee MDPI, Basel, Switzerland.
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distributed under the terms and
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Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
1Centre for Mountain Futures (CMF), Kunming Institute of Botany, Honghe County 654400, China;
6371105502@lamduan.mfu.ac.th (D.B.); peter@mail.kib.ac.cn (P.E.M.)
2Innovative Institute of Plant Health, Zhongkai University of Agriculture and Engineering, Haizhu District,
Guangzhou 510225, China
3Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand;
shike.hua13@lamduan.mfu.ac.th
4CIFOR-ICRAF China Program, World Agroforestry Centre, Kunming 650201, China
5School of Life Science and Technology, University of Electronic Science and Technology of China,
Chengdu 611731, China; sajeewa@uestc.edu.cn
6No. 128/1-J, Azad Housing Society, Curca, P.O., Goa Velha 403108, India; bhatdj@gmail.com
7Research Center of Microbial Diversity and Sustainable Utilization, Chiang Mai University,
Chiang Mai 50200, Thailand; saisamorn.l@cmu.ac.th
8Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand
9Academy of Science, The Royal Society of Thailand, Bangkok 10300, Thailand
10 CAS Key Laboratory for Plant Biodiversity and Biogeography of East Asia (KLPB),
Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China
*Correspondence: wanasinghe@mail.kib.ac.cn (D.N.W.); kevin.hyd@mfu.ac.th or
kdhyde3@gmail.com (K.D.H.)
Abstract: An undetermined saprobic fungal taxon from Yunnan (China) is revealed as a new genus
in Scortechiniaceae (Coronophorales). The novel taxon, Yuxiensis, is characterized by immersed to
erumpent, semi-globose ascomata, which are not surrounded by any tomentum or conspicuous
subiculum, a subcylindrical quellkörper in the centrum, clavate asci with long pedicels and allantoid
hyaline ascospores with granular contents. Maximum likelihood and Bayesian posterior probability
analyses based on LSU, ITS, tef1 and rpb2 sequence data depict a close phylogenetic relationship of the
new genus to Pseudocatenomycopsis, hence, confirming its placement in Scortechiniaceae.Parasympodiel-
laceae, thus far belonging to Parasympodiellales, is transferred to Coronophorales based on multi-gene
phylogenetic evidence. Additionally, the incertae sedis monotypic genus Arthrocristula is treated as a
synonym of Parasympodiella, with Arthrocristula hyphenata recombined as Parasympodiella hyphenata
comb. nov., as the type strain of Arthrocristula hyphenata clusters inside the Parasympodiellaceae clade
along with other Parasympodiella taxa.
Keywords: 2 new taxa; 1 new combination; Hypocreomycetidae; phylogeny; Sordariomycetes; Yunnan
1. Introduction
Members of Coronophorales are generally found in terrestrial habitats. These fungi
occur as saprobes on woody substrates, with no specific host per se, and their diver-
sity spans tropical and temperate regions [
1
–
4
]. The order is accommodated in Hypocre-
omycetidae and it includes six families, viz. Bertiaceae,Ceratostomataceae,Chaetosphaerellaceae,
Coronophoraceae,Nitschkiaceae and Scortechiniaceae, classified based on molecular data and
morphology [2,5–8].
Life 2021,11, 1011. https://doi.org/10.3390/life11101011 https://www.mdpi.com/journal/life

Life 2021,11, 1011 2 of 17
Scortechiniaceae was introduced by Huhndorf et al. [
9
] to accommodate three saprobic
genera, Euacanthe,Neofracchiaea and Scortechinia using morphological characteristics and
LSU sequence data. Mugambi and Huhndorf [
2
] later added six additional genera, Bicil-
iospora,Coronophorella,Cryptosphaerella,Scortechiniella,Scortechiniellopsis and Tympanopsis
to the family. All genera formed a well-supported clade in a multi-gene phylogeny based
on LSU, tef1 and rpb2 sequence data [
9
]. Interestingly, all these taxa contain a distinctive
phenotypic character in their centrum, namely, the quellkörper, which demarcates them
from taxa belonging to other families in the order [
2
,
9
]. The quellkörper has been described
as a subcylindrical to inverted-conical structure located in the centrum, made up of a mass
of thick-walled, hyaline cells tightly arranged in a circular manner [
2
,
9
,
10
]. It is usually
attached to the roof of the ascoma, orienting downward, and can extend until the base
of the sporocarp [
2
,
9
,
10
]. The structure is mainly thought to be involved in ascospore
discharge [
2
,
9
,
10
]. The quellkörper has, over the years, been considered and justified by
taxonomists as the primary family-level character [2,3,9].
The monotypic taxon, Pseudocatenomycopsis, was then recently added to Scortechiniaceae
based on LSU and ITS sequence data. The genus is only known in its asexual morph [
11
].
Latest taxonomic revisions of the family conducted by Huang et al. [
12
] revealed that
Cryptosphaerella had to be transferred to Sordariomycetes genera incertae sedis based on
the morphology of its type. The taxa Neocryptosphaerella and Pseudocryptosphaerella have
instead been introduced to accommodate the Cryptosphaerella taxa having sequence data in
GenBank at present [12].
Another order, Parasympodiellales, belonging to the same subclass, Hypocreomycetidae,
was established by Hernández-Restrepo et al. [
13
], along with its type family, Parasympodiel-
laceae, to accommodate Parasympodiella, whose taxa formed a monophyletic clade in their
phylogenetic tree. Members of Parasympodiellales are only known in their asexual morphs
and are mainly characterized by sympodial conidiogenous cells and thallic-arthric coni-
dia [
13
–
16
]. Furthermore, they usually have a stylaspergillus-like synasexual morph [
13
,
16
].
In the multi-gene phylogenetic analyses conducted by Hyde et al. [
8
], it was observed that
Parasympodiellales clustered as sister to Scortechiniaceae (Coronophorales), following which
the authors mentioned the status of this order requiring further revision.
Similarly, many species of Sordariomycetes still require taxonomic revision, especially
those which have so far been identified based on morphology alone. Such is the case for
Arthrocristula hyphenata, belonging to the monotypic genus Arthrocristula [
17
], currently
placed in Ascomycota genera incertae sedis [
18
]. DNA sequence data for A. hyphenata were
recently retrieved by Vu et al. [
19
] and deposited in GenBank. With both the morphology
and molecular data for the taxon now available, it becomes largely feasible to determine
the correct placement of A. hyphenata.
The present study aims to introduce a new fungal genus, Yuxiensis, in Scortechiniaceae,
collected on a woody host in Yuxi (Yunnan, China) based on morphology and phylogenetic
analyses of combined LSU–ITS–tef1–rpb2 sequence data. Furthermore, Parasympodiellaceae
is included in Coronophorales based on phylogenetic evidence. Arthrocristula is synonymized
under Parasympodiella, with Arthrocristula hyphenata combined as Parasympodiella hyphenata
Bundhun & K.D. Hyde, comb. nov., and is accommodated in Parasympodiellaceae based on
DNA sequence data analyzed in the present study.
2. Materials and Methods
2.1. Specimen Collection and Morphological Studies
Dead twigs of an undetermined deciduous host were collected from Yuxi, Yunnan
Province, China, during the dry season in May 2019. The samples were taken to the my-
cology laboratory at the Kunming Institute of Botany, in a plastic Ziploc bag and stored
inside a paper envelope. External examinations were made using a Motic SMZ 168 Series
stereo-microscope. Morphological characters were examined by hand sectioning of sporo-
carps and placed on water-mounted glass slides. Microscopic photography was conducted
using a Nikon ECLIPSE 80i compound microscope (Nikon, Tokyo, Japan) fitted with a

Life 2021,11, 1011 3 of 17
Canon EOS 600D camera. The structures which were observed and measured include
diameter, height, shape, and color of ascomata; ascomatal wall’s width and cell structure;
quellkörper shape; shape, length, and width of asci and shape, size, and ornamentation
of ascospores. The Tarosoft (R) Image Frame Work version 0.9.7. program was used for
the measurements of photomicrograph structures. Images used for figures were processed
with the Adobe Photoshop CS6 Extended version 13.0.1 software (Adobe Systems, San Jose,
California). The holotype was deposited in the herbarium of Cryptogams Kunming Insti-
tute of Botany Academia Sinica (HKAS). Both Facesoffungi [
20
] and Index Fungorum [
21
]
numbers were obtained.
2.2. DNA Extraction, PCR Amplification and Sequencing
No culture could be obtained for the collected sample despite several trials on various
media, including malt extract agar, potato dextrose agar, corn meal agar, or water agar
under different incubation conditions. Therefore, DNA was extracted directly from the
fruiting bodies of the fungus as outlined by Wanasinghe et al. [22].
According to the manufacturer’s instructions, DNA was extracted from fresh sporo-
carps using the Biospin Fungus Genomic DNA Extraction Kit-BSC14S1 (BioFlux, P.R.
China). Primers LR0R/LR5 [
23
], ITS5/ITS4 [
24
], EF1-983F/EF1-2218R [
25
], and fRPB2-
5f/fRPB2-7cR [
26
,
27
] were used to amplify the DNA sequences of the partial 28S large
subunit rDNA (LSU), internal transcribed spacers (ITS), translation elongation factor 1-
α
(tef1), and RNA polymerase II second largest subunit (rpb2). The total volume of 25
µ
L
contained 12.5
µ
L of 2
×
PCR Master Mix with dye (0.1 U Taq Polymerase/
µ
L, 500
µ
m
dNTP each), 20 mM Tris-HCl (pH 8.3, 100 mM KCl, 3 mM MgCl
2
), 1
µ
L of each primer,
9.5 µL of double-distilled water, and 1 µL (100–500 ng) of DNA template.
The PCR protocols were programmed as described in Wanasinghe et al. [
28
]. The PCR
products were verified by staining with ethidium bromide on 1% agarose electrophoresis
gels. They were then purified according to the company protocols and DNA sequencing
was performed at Shanghai Sangon Biological Engineering Technology & Services Co.
(Shanghai, P.R. China). Forward and reverse DNA sequence data were obtained and
analyzed. Consensus sequences were generated using the SeqMan software (DNAStar,
Inc., Madison, WI, USA). The newly acquired sequence data from the present study were
deposited in GenBank for subsequent studies [29] (Table 1).
Table 1.
Taxa used in this study and their corresponding GenBank accession numbers. Generated sequence data for the new
taxon are in bold.
Taxa Strains
GenBank Accession Numbers
LSU ITS tef1 rpb2
Acanthonitschkea argentinensis SMH1395 AY695259 - FJ969042 FJ968943
Acanthonitschkea tristis SMH4723 FJ968949 - FJ969043 -
Bertia didyma SMH4719 FJ968958 - - -
Bertia ngongensis GKM1239 * FJ968954 - - -
Bertia moriformis SMH4320 AY695260 - - AY780151
Bertia orbis GKM1271 FJ968955 - FJ969009 -
Biciliospora velutina GKM1268 FJ968964 - FJ969018 FJ968932
Chaetosphaerella fusca GKML124N FJ968967 - FJ969002 -
Chaetosphaerella phaeostroma SMH4257 AY695264 - FJ969004 FJ968940
Coronophora gregaria ANM1555 - - FJ969007 FJ968938
Coronophorella chaetomioides GKM1099 FJ968969 - FJ969034 FJ968922
Crassochaeta nigrita SMH1667 AY695265 - - -
SMH2931 AY695266 - - -
Emericellopsis alkalina CBS 127350 * MH875970 MH864534 KC998993 KC999029

Life 2021,11, 1011 4 of 17
Table 1. Cont.
Taxa Strains
GenBank Accession Numbers
LSU ITS tef1 rpb2
Euacanthe usambarensis GKM1221 FJ968978 - FJ969026 FJ968927
SMH4408 AY695267 - - -
Fracchiaea broomeana
SMH347 FJ968979 - FJ969041 FJ968947
SMH2809 AY695268 - FJ969039 FJ968942
GKM1071 - - FJ969040 FJ968919
Fracchiaea myricoides IFRD 9201 * KX856174 KX856173 - -
HKAS 115760 MZ713199 MZ713184 MZ712579 MZ712580
Fracchiaea lunata GKM1089 - - - FJ968921
Fracchiaea sp. GKM1250 - - FJ969005 -
Gaillardiella pezizoides GKM1245 FJ968981 - FJ969006 -
Melanospora tiffanii ATCC 15515 AY015630 - - AY015637
SMCD2222 FJ748915 FJ748921 - -
Melanospora zamiae ATCC 12340 AY046579 - - AY046580
ATCC 96173 AY057906 - - -
Neocryptosphaerella celata GKM1231 * FJ968975 - FJ969035 FJ968929
Neocryptosphaerella globosa
GKM471N * FJ968977 - FJ969036 FJ968935
GKM437N - - FJ969038 -
GKM414N - - FJ969037 -
396N FJ968976 - - -
Neofracchiaea callista SMH2689 AY695269 - FJ969020 FJ968941
Nitschkia calyculus SMH918 FJ968983 - - -
Nitschkia grevillei SMH4663 AY346294 - - -
Nitschkia meniscoidea SMH1523 * AY695270 - - -
Nitschkia tetraspora GKML148N FJ968987 - FJ969011 FJ968936
SMH4787 FJ968984 - FJ969010 -
Parasympodiella elongata
CBS:522.93 * GQ303314 GQ303283 - -
CBS:124768 GQ303311 GQ303280 - -
CPC:13288 GQ303312 GQ303281 - -
CPC:13498 GQ303313 GQ303282 - -
Parasympodiella eucalypti CBS:124767 * GQ303315 GQ303284 - -
LCM 815.01 - MF495381 - -
Parasympodiella hyphenata CBS 583.82 * MH873274 MH861530 - -
Parasympodiella lauri FMR_13132 * KY853518 KY853457 - -
Parasympodiella laxa CBS 102698 GQ303316 GQ303285 - -
Parasympodiella longispora CBS 544.84 * MH873476 MH861778 - -
KACC 41225 - GQ272636 - -
Pseudocatenomycopsis rothmanniae CPC 22733 * KF777237 KF777185 - -
Pseudocryptosphaerella costaricensis MO2111 * FJ968971 - FJ969028 -
Pseudocryptosphaerella cylindriformis
GKM434N * FJ968972 - FJ969031 FJ968934
GKM1187 GQ217531 - FJ969033 FJ968925
GKM1042 FJ968973 - FJ969032 FJ968918
GKM1075 - - FJ969030 FJ968920

Life 2021,11, 1011 5 of 17
Table 1. Cont.
Taxa Strains
GenBank Accession Numbers
LSU ITS tef1 rpb2
Pseudocryptosphaerella elliptica SMH4722 * FJ968974 - FJ969029 FJ968944
Pseudocryptosphaerella malindensis GKM1150 * FJ968970 - FJ969027 FJ968923
Pseudohyaloseta pandanicola MFLUCC 16-0316 * MH376737 MH388363 MH388398 MH412733
Scortechinia acanthostroma
SMH1164 FJ968989 - FJ969014 FJ968924
SMH1143 FJ968988 - FJ969012 FJ968948
GKML163N FJ968991 - FJ969015 -
SMH5313 FJ968990 - FJ969013 -
Scortechinia diminuspora SMH4763 * FJ968992 - - -
HUEFS:194245 KT003703 - - -
Scortechiniella similis SMH2006 FJ968994 - FJ969019 FJ968945
Scortechiniellopsis leonensis GKM1269 FJ968993 - FJ969021 FJ968933
Stachybotrys microspora KLM 3-2 KU760387 KU760377 KU760392 KU760397
Tympanopsis confertula
GKM1242 FJ968997 - FJ969023 FJ968930
SMH1567 FJ969001 - - FJ968939
SMH4841 FJ968998 - FJ969024 FJ968946
SMH2648 AY695272 - - -
Tympanopsis uniseriata GKM1203 FJ968999 - FJ969016 FJ968926
GKM1228 FJ969000 - FJ969017 -
Yuxiensis granularis HKAS 109580 * MZ713198 MZ713183 MZ712577 MZ712578
Type strains are indicated in ‘*’.
2.3. Phylogenetic Analyses
Verified sequences were initially used for BLASTn analyses, following which closely
related sequences were downloaded from GenBank based on BLAST similarities and rele-
vant publications [
2
,
8
] (Table 1). Alignment of each locus was performed using MUSCLE
in MEGA X (Molecular Evolutionary Genetics Analysis), using default conditions for gap
openings and gap extension penalties. It was then improved whenever necessary in the
BioEdit v.7.0.5.2 software [30].
Maximum likelihood (ML) and Bayesian posterior probability (BYPP) analyses were
conducted using both individual and combined datasets. Prior to ML analysis, the sequence
alignments were converted from FASTA into PHYLIP format using the ALTER (alignment
transformation environment, http://www.singgroup.org/ALTER/, accessed on 30 August
2021) bioinformatics web tool [
31
]. They were then used to generate ML trees using
RAxML-HPC2 on XSEDE (v.8.2.10) [
32
] with the GTRGAMMA substitution model and
bootstrapping with 1000 replicates.
The BYPP analysis was generated using Markov Chain Monte Carlo sampling in
MrBayes v3.1.2 [33,34]. MrModeltest v.2.3 [35] was used to estimate the best evolutionary
model for each gene region under the Akaike Information Criterion (AIC) implemented in
PAUP v.4.0b10 [
36
]. The best-fit model was determined as GTR+I+G for LSU, tef1, and rpb2
while GTR+G for ITS. Six simultaneous Markov chains were run for 4.5M generations with
trees sampled every 100th generation. The first 20% of generated trees were the burn-in
phase and discarded. The remaining 80% of trees were used to calculate posterior probabil-
ities in the majority rule consensus tree. Phylograms were configured in FigTree v.1.4.0 [
37
]
and modified in Microsoft PowerPoint (2013). The final alignments and phylogenetic tree
were deposited in TreeBASE, submission ID: 28713 (http://www.treebase.org/ , accessed
on 30 August 2021).

Life 2021,11, 1011 6 of 17
3. Results
3.1. Phylogenetic Analyses
The final concatenated LSU–ITS–tef1–rpb2 alignment (Figure 1) comprised 75 strains
including the outgroup taxa Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pandan-
icola (MFLUCC 16-0316), and Stachybotrys microspora (KLM 3-2). The manually adjusted
dataset consisted of 3845 characters including gaps (LSU: 1086, ITS: 768, tef1: 813, rpb2:
1178). The best scoring RAxML tree with final optimization had a likelihood value
of
−
41,633.775219. The matrix had 2266 distinct alignment patterns, with 49.47% of
gaps and completely undetermined characters. Estimated base frequencies were as fol-
lows:
A = 0.236728
,
C = 0.275120
, G = 0.286267, T = 0.201885, with substitution rates
AC = 1.265283
,
AG = 3.588580
, AT = 1.636499, CG = 1.073052, CT = 7.584875, GT = 1.000000.
The gamma distribution shape parameter α= 0.342349 and Tree-length = 7.680873.
Life 2021, 11, x FOR PEER REVIEW 6 of 19
3. Results
3.1. Phylogenetic Analyses
The final concatenated LSU–ITS–tef1–rpb2 alignment (Figure 1) comprised 75 strains
including the outgroup taxa Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pandan-
icola (MFLUCC 16-0316), and Stachybotrys microspora (KLM 3-2). The manually adjusted
dataset consisted of 3845 characters including gaps (LSU: 1086, ITS: 768, tef1: 813, rpb2:
1178). The best scoring RAxML tree with final optimization had a likelihood value of -
41,633.775219. The matrix had 2266 distinct alignment patterns, with 49.47% of gaps and
completely undetermined characters. Estimated base frequencies were as follows: A =
0.236728, C = 0.275120, G = 0.286267, T = 0.201885, with substitution rates AC = 1.265283,
AG = 3.588580, AT = 1.636499, CG = 1.073052, CT = 7.584875, GT = 1.000000. The gamma
distribution shape parameter α = 0.342349 and Tree-length = 7.680873.
Figure 1. Cont.

Life 2021,11, 1011 7 of 17
Life 2021, 11, x FOR PEER REVIEW 7 of 19
Figure 1. Phylogram generated from maximum likelihood (RAxML) based on LSU–ITS–tef1–rpb2 matrix for Coronopho-
rales. The tree is rooted with Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pandanicola (MFLUCC 16-0316) and Stachy-
botrys microspora (KLM 3-2). Maximum likelihood bootstrap (≥65) and BYPP (≥0.95) supports are shown, respectively,
above or below the branches. Type strains are in bold while novelty and the recombined taxon are in blue.
The families Bertiaceae, Ceratostomataceae, Chaetosphaerellaceae, Coronophoraceae, Nitsch-
kiaceae, Parasympodiellaceae and Scortechiniaceae grouped together, each forming a mono-
phyletic clade in the ML tree (Figure 1). The tree topology resulting from the BYPP anal-
ysis mainly differed from the ML one with regard to the placement of Coronophora gregaria
(ANM1555) (Coronophoraceae), which clustered within Parasympodiellaceae (Figure A1).
Parasympodiellaceae formed a sister clade with Scortechiniaceae with statistical support
of 96% ML, 1.00 BYPP. It comprised ten strains of Parasympodiella and a strain of ‘Ar-
throcristula hyphenata’ (CBS 583.82), which clustered together with 98% ML and 0.95 BYPP
statistical support (Figure 1).
Our strain HKAS 109580 formed a distinct lineage within Scortechiniaceae, placed sis-
ter to Pseudocatenomycopsis rothmanniae (CPC 22733) with low statistical support (Figure
1).
3.2. Taxonomy
In this section, the amended descriptions and notes for Coronophorales, Parasympodiel-
laceae, and Parasympodiella are given. Furthermore, descriptions, notes, and illustrations
are given for the following taxa: Parasympodiella hyphenata comb. nov. and Yuxiensis gran-
ularis gen. et sp. nov.
Figure 1.
Phylogram generated from maximum likelihood (RAxML) based on LSU–ITS–tef1–rpb2 matrix for Coronophorales
.
The tree is rooted with Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pandanicola (MFLUCC 16-0316) and Stachybotrys
microspora (KLM 3-2). Maximum likelihood bootstrap (
≥
65) and BYPP (
≥
0.95) supports are shown, respectively, above or
below the branches. Type strains are in bold while novelty and the recombined taxon are in blue.
The families Bertiaceae,Ceratostomataceae,Chaetosphaerellaceae,Coronophoraceae,Nitschki-
aceae,Parasympodiellaceae and Scortechiniaceae grouped together, each forming a mono-
phyletic clade in the ML tree (Figure 1). The tree topology resulting from the BYPP analysis
mainly differed from the ML one with regard to the placement of Coronophora gregaria
(ANM1555) (Coronophoraceae), which clustered within Parasympodiellaceae (Figure A1).
Parasympodiellaceae formed a sister clade with Scortechiniaceae with statistical support of
96% ML, 1.00 BYPP. It comprised ten strains of Parasympodiella and a strain of ‘Arthrocristula
hyphenata’ (CBS 583.82), which clustered together with 98% ML and 0.95 BYPP statistical
support (Figure 1).
Our strain HKAS 109580 formed a distinct lineage within Scortechiniaceae, placed sister
to Pseudocatenomycopsis rothmanniae (CPC 22733) with low statistical support (Figure 1).
3.2. Taxonomy
In this section, the amended descriptions and notes for Coronophorales,Parasympodiel-
laceae, and Parasympodiella are given. Furthermore, descriptions, notes, and illustrations are
given for the following taxa: Parasympodiella hyphenata comb. nov. and Yuxiensis granularis
gen. et sp. nov.

Life 2021,11, 1011 8 of 17
3.2.1. Coronophorales Nannf., Nova Acta R. Soc. Scient. Upsal., Ser. 4 8(no. 2): 54 (1932)
Amend
Index Fungorum number: IF 501516; Facesoffungi number: FoF 06517
=Parasympodiellales Hern.-Restr., Gené, R.F. Castañeda & Crous 2017
Saprobic on leaves, wood, or associated with sclerotia. Sexual morph: see Mugambi and
Huhndorf [
2
], Hyde et al. [
8
], Nannfeldt [
10
]. Asexual morph: Hyphomycetous. Conidio-
phores simple or branched, septate, straight to flexuous, brown, smooth or comprising rough
swellings along the whole length in some genera or geniculations in others. Conidiogenous
cells apical, lateral or intercalary, usually hyaline, often sympodial, blastic or thallic. Conidia
hyaline or brown, aseptate to septate, globose, oval, elliptical, ellipsoidal, pyriform, cylin-
drical, oblong or spindle-shaped, smooth or verrucose, solitary or produced in branched
or unbranched chains. Synasexual morph: stylaspergillus-like (see Hernández-Restrepo
et al. [
13
], Cheewangkoon et al. [
16
]), or arthrocristula-like: Conidiophores branched, brown,
smooth. Conidiogenous cells apical, lateral, sympodial, thallic. Conidia pale brown or brown,
aseptate to septate, cylindrical to oblong, produced in unbranched chains.
Type family: Coronophoraceae Höhn.
Notes: The phylogenetic analyses based on the combined LSU–ITS–tef1–rpb2 se-
quence data, conducted in the present study, supports the inclusion of Parasympodiellales
in Coronophorales, as Parasympodiellaceae is sister to Scortechiniaceae with 96% ML and 1.00
BYPP statistical support within the order (Figure 1). The description for the asexual morph
of taxa in Coronophorales is therefore emended to include the morphological characteristics
of Parasympodiellaceae.
3.2.2. Parasympodiellaceae Hern.-Restr., Gené, Guarro & Crous, in Hernández-Restrepo,
Gené, Castañeda-Ruiz, Mena-Portales, Crous & Guarro, Stud. Mycol. 86: 87 (2017) Amend
Index Fungorum number: IF 820298; Facesoffungi number: FoF 06518
Saprobic on leaves and twigs or associated with sclerotia. Sexual morph: Undetermined.
Asexual morph: Hyphomycetous. Conidiophores micro- to macronematous, brown, usually
unbranched, septate. Conidiogenous cells thallic, terminal or intercalary, sympodial. Conidia
thallic-arthric, aseptate or septate, cylindrical, hyaline, often in chains, with schizolytic
secession. Synasexual morph: stylaspergillus-like (see Hernández-Restrepo et al. [
13
],
Cheewangkoon et al. [
16
]) or arthrocristula-like: Conidiophores micro- to macronematous,
brown, branched. Conidiogenous cells thallic, terminal, lateral, sympodial. Conidia thallic-
arthric, aseptate or septate, cylindrical to oblong, pale brown or brown, generally in chains,
with rhexolytic secession.
Type genus: Parasympodiella Ponnappa
Notes: Parasympodiellaceae is now included in Coronophorales, which at present com-
prises seven families (this study). This family, in turn, accommodates the single genus
Parasympodiella.
3.2.3. Parasympodiella Ponnappa Trans. Br. Mycol. Soc. 64(2): 344 (1975) Amend
Index Fungorum number: IF 9226; Facesoffungi number: FoF 05188
=Arthrocristula Sigler, M.T. Dunn & J.W. Carmich., Mycotaxon 15: 409 (1982)
Saprobic on leaves and twigs or associated with sclerotia. Sexual morph: Undeter-
mined. Asexual morph: Hyphomycetous. Conidiophores micro- to macronematous, brown,
generally unbranched, septate, straight to geniculate. Conidiogenous cells thallic, terminal
or intercalary, indeterminate, sympodial, unbranched. Conidia thallic-arthric, produced
in unbranched chains, cylindrical, aseptate or septate, hyaline, seceding schizolytically,
often with a septal plug. Synasexual morph: stylaspergillus-like (see Hernández-Restrepo
et al. [
13
], Cheewangkoon et al. [
16
]) or arthrocristula-like: Conidiophores micro- to macrone-
matous, brown, branched. Conidiogenous cells thallic, terminal, lateral, indeterminate,
sympodial, unbranched. Conidia thallic-arthric, produced in unbranched chains, aseptate
or septate, pale brown or brown, cylindrical to oblong, with rhexolytic secession, with frills
of remnant cells at each end.

Life 2021,11, 1011 9 of 17
Type species: Parasympodiella laxa (Subram. & Vittal) Ponnappa
Notes: With the exact taxonomic placement being uncertain, the hyphomycetous
taxon Arthrocristula has so far been maintained in Ascomycota genera incertae sedis [
18
,
38
].
In the present study, the type strain of A. hyphenata (CBS 583.82) was found to cluster
within Parasympodiellaceae, indicating a close phylogenetic affinity to Parasympodiella (Fig-
ure 1). Morphologically, the conidiophores of Parasympodiella are unbranched or sparingly
branched, thick-walled, and brown, becoming paler towards the conidiogenous regions.
They are often geniculate, with terminal or intercalary sympodial conidiogenous cells,
which are mostly thallic. The secession of conidia occurs schizolytically [
13
,
14
,
16
]. The
conidiophores of Arthrocristula are, however, well-branched with no conspicuous genic-
ulation and are initially hyaline, becoming thick-walled and brown upon maturity. The
conidiogenous cells secede rhexolytically into arthroconidia, leaving the mature conidia
with remnants of the separating cells at each end with small frills. The terminal cells of the
conidiogenous hyphae remain as empty cells [
17
]. Considering these morphological differ-
ences, the two genera can be considered as distinct. However, given that Parasympodiella
has a stylaspergillus-like synasexual morph, the fact that Arthrocristula also is another
synasexual morph of Parasympodiella cannot be ruled out. Fungi have been reported to
have two or more morphologically distinct asexual morphs [
39
–
41
]. Moreover, phylogeny
supports the inclusion of Arthrocristula in Parasympodiella (Figure 1). Therefore, the former
is synonymized under Parasympodiella in the present study.
3.2.4. Parasympodiella hyphenata (Sigler, M.T. Dunn & J.W. Carmich.) Bundhun & K.D.
Hyde, comb. nov.
Index Fungorum number: IF 558677; Facesoffungi number: FoF 10183, Figure 2.
≡
Arthrocristula hyphenata Sigler, M.T. Dunn & J.W. Carmich., Mycotaxon 15: 409
(1982)
Associated with sclerotia of Sclerotinia minor in soil. Sexual morph: Undetermined.
Asexual morph: Hyphomycetous. Mycelium
in vitro
comprises narrow, septate vegetative
hyphae (2–3
µ
m diam.), hyaline when immature and brown at maturity. Conidiophores
originating from vegetative hyphae, undifferentiated, initially hyaline, turning brown and
thick-walled when mature, branched, narrow at first, widening and extending apically and
laterally after that to give rise to fertile conidiogenous hyphae. Conidiogenous cells (80–100
×
4–5.5
µ
m), delimited by a basal septum, developing successively, generally unbranched,
indeterminate, initially non-septate, hyaline, becoming randomly septate and pigmented
in alternate cells on maturity, undergoing rhexolytic secession to give rise to arthroconidia.
Conidia (5–12
×
6–7
µ
m), thallic-arthric, brown, aseptate to septate, cylindrical to oblong,
formed in a chain with hyaline separating cells, with fragments of intervening cell walls
remaining attached to both ends of conidium after secession, terminal cells of the chains
remain empty. Chlamydospores
in vitro
(12–16
×
8–13
µ
m), thick-walled, brown, terminal
and intercalary (adapted from Sigler et al. [17]).
Notes: The type and single taxon of Arthrocristula,A. hyphenata is combined under
Parasympodiella following the synonymy of Arthrocristula under Parasympodiella in the
present study. Parasympodiella hyphenata comb. nov. has been reported from the sclerotium
of Sclerotinia minor buried in the soil [
17
]. It is mainly characterized by branched coni-
diophores and alternatively pigmented conidiogenous cells, which produce conidia by
seceding rhexolytically [17].

Life 2021,11, 1011 10 of 17
Life 2021, 11, x FOR PEER REVIEW 10 of 19
Figure 2. Parasympodiella hyphenata redrawn from Sigler et al. [17] and Seifert et al. [42]. (a) Conidiogenous hyphae origi-
nating from a conidiophore. (b,c) Conidiogenous hyphae, with rhexolytically dehiscing intervening cells (shown by ar-
rows) and darker pigmented area denoting conidia. (d–f) Arthroconidia with remnants of cell walls attached at both ends.
Scale bars: (a–c) = 30 μm, (d) = 5 μm, (e,f) = 10 μm (scale bars adapted based on original description in Sigler et al. [17]).
3.2.5. Yuxiensis Bundhun & K.D. Hyde, gen. nov.
Index Fungorum number: IF 558675; Facesoffungi number: FoF 10184
Etymology–Referring to the city Yuxi in China.
Saprobic on dead wood. Sexual morph: Ascomata immersed to erumpent, appearing
superficial on worn off substrate, astromatic to stromatic, aggregated, black, non-ostiolate,
semi-globose when fresh, collapsing when dry, without hair or bristles, subiculum incon-
spicuous, coriaceous. Ascomatal wall comprising 2–3 types of layers; outermost layer heav-
ily pigmented, composed of thick-walled, very dark brown cells, inner layer comprising
Figure 2.
Parasympodiella hyphenata redrawn from Sigler et al. [
17
] and Seifert et al. [
42
]. (
a
) Conidio-
genous hyphae originating from a conidiophore. (
b
,
c
) Conidiogenous hyphae, with rhexolytically
dehiscing intervening cells (shown by arrows) and darker pigmented area denoting conidia. (
d
–
f
)
Arthroconidia with remnants of cell walls attached at both ends. Scale bars: (
a
–
c
) = 30
µ
m,
(d)=5µm
,
(e,f) = 10 µm (scale bars adapted based on original description in Sigler et al. [17]).
3.2.5. Yuxiensis Bundhun & K.D. Hyde, gen. nov.
Index Fungorum number: IF 558675; Facesoffungi number: FoF 10184
Etymology–Referring to the city Yuxi in China.
Saprobic on dead wood. Sexual morph: Ascomata immersed to erumpent, appearing
superficial on worn off substrate, astromatic to stromatic, aggregated, black, non-ostiolate,
semi-globose when fresh, collapsing when dry, without hair or bristles, subiculum in-
conspicuous, coriaceous. Ascomatal wall comprising 2–3 types of layers; outermost layer
heavily pigmented, composed of thick-walled, very dark brown cells, inner layer compris-
ing thick-walled cells of textura globulosa to textura angularis, innermost layer composed
of flattened, thin-walled cells of textura prismatica toward the locule; Munk pores visible,
few per cell. Hamathecium composed of large, subcylindrical quellkörper, attached to the
roof of the centrum and extending until base of the ascoma; paraphyses indistinct. Asci
8-spored, unitunicate, clavate, long-pedicellate, rounded at apex, lacking an apical ring,
thin-walled, evanescent. Ascospores irregularly arranged, cylindrical to allantoid, hyaline,
aseptate, with granular contents, lacking mucilaginous sheath or appendage. Asexual
morph: Undetermined.
Type: Yuxiensis granularis Bundhun, Wanas. & K.D. Hyde

Life 2021,11, 1011 11 of 17
Notes: Yuxiensis is introduced in Scortechiniaceae as a new quellkörper-bearing taxon
distinct from all other genera in the family, based on LSU, ITS, tef1 and rpb2 sequence data.
The new genus has a close phylogenetic affinity to Pseudocatenomycopsis even though this
relationship is not statistically significant (Figure 1). This low support may possibly be
accounted for by insufficient taxon sampling. Nevertheless, the new taxon being intro-
duced in the present study constantly clusters with Pseudocatenomycopsis in all phylogenies
(single, not shown; and concatenated, Figure 1). The asexual morph for Yuxiensis could
not be obtained in the present study. It thus cannot be morphologically compared with
Pseudocatenomycopsis, which has been described in its asexual morph only. Pseudocatenomy-
copsis has been introduced from Zambia as a saprobe on the stem of Rothmannia engleriana
(Rubiaceae) [
11
]. The new genus is also phylogenetically close to Euacanthe (Figure 1). It
morphologically differs from Euacanthe in terms of ascomatal position and surface as well
as ascospore ornamentation [12,43].
3.2.6. Yuxiensis granularis Bundhun, Wanas. & K.D. Hyde, sp. nov.
Index Fungorum number: IF 558676; Facesoffungi number: FoF 10185, Figure 3.
Etymology—The specific epithet refers to the granular contents of the ascospores.
Holotype–HKAS 109580
Saprobic on dead twigs of deciduous hosts in terrestrial habitats. Sexual morph:
Ascomata 250–400
µ
m high, 550–700
µ
m diam. (
x
= 309
×
642
µ
m, n = 5), astromatic to
stromatic, immersed to erumpent, appearing superficial after substrate has worn away,
aggregated, black, non-ostiolate, semi-globose when fresh, collapsing upon drying, without
hair or bristles, with inconspicuous subiculum, coriaceous. Ascomatal wall made up of 2–3
layers, almost equally thickened, 40–70
µ
m wide at the apex and base, 50–70
µ
m wide
at the sides; heavily pigmented at outermost layer, composed of thick-walled, very dark
brown cells, with inner layer comprising thick-walled cells (25–45
µ
m) of textura globulosa
to textura angularis, innermost layer composed of flattened, thin-walled cells of textura
prismatica toward the locule; Munk pores visible, few per cell. Hamathecium made up of
subcylindrical quellkörper, 290
µ
m long and 220
µ
m wide. Asci 40–80
×
5–10
µ
m (
x
= 55.8
×
8.4
µ
m, n = 10), spore bearing part 15–30
µ
m, pedicel 15–50
µ
m, 8-spored, unitunicate,
clavate, long-pedicellate, rounded at apex, lacking an apical ring, thin-walled, evanescent.
Ascospores 8–15
×
2–3
µ
m (
x
= 11.7
×
2.3
µ
m, n = 35), irregularly arranged, cylindrical
to allantoid, hyaline, unicellular, aseptate, with granular contents, lacking mucilaginous
sheath or appendage. Asexual morph: Undetermined.
Material examined: CHINA, Yunnan, Yuxi, Yi and Dai Autonomous County, Yuanjiang
Hani, 23.74074 N, 102.17735 E, on woody litter of an undetermined deciduous host, 24 May
2019, 1345 msl, D.N. Wanasinghe, DW0636-19 (HKAS 109580, holotype).
Notes: In the multi-gene phylogeny, Yuxiensis granularis is more closely related to Pseu-
docatenomycopsis rothmanniae, followed by Euacanthe usambarensis (=Euacanthe foveolata [
12
])
(Figure 1). The LSU sequence of Yuxiensis granularis is 95% similar to Pseudocatenomycopsis
rothmanniae (GenBank KF777237; similarity = 869/910(95%), Gaps = 3/910(0%)). The ITS
sequence of Yuxiensis granularis is 85% similar to Pseudocatenomycopsis rothmanniae (Gen-
Bank KF777185; similarity = 470/552(85%), Gaps = 10/552(1%)). Pseudocatenomycopsis
rothmanniae has only LSU and ITS sequence data deposited in GenBank, and hence the
protein-coding genes, tef1 and rpb2 could not be compared. Morphological comparison
between the two taxa is currently unfeasible since the single species of Pseudocatenomycopsis,
P. rothmanniae, has been introduced in its asexual morph [
11
], while the asexual morph for
Yuxiensis granularis could not be obtained.
Yuxiensis granularis resembles Euacanthe usambarensis in having ascomata which be-
come collabent on drying and 8-spored, clavate asci [
2
,
12
,
43
]. However, the new species
has immersed to erumpent, glabrous ascomata that are not surrounded by any conspicuous
subiculum or tomentum, whereas Euacanthe usambarensis comprises superficial ascomata
with a setose surface, sitting on a dense subiculum [12,43]. The asci of Yuxiensis granularis
are long-pedicellate whereas those of Euacanthe usambarensis have short pedicels [
12
,
43
].

Life 2021,11, 1011 12 of 17
Furthermore, while the ascospore contents of Yuxiensis granularis are granular, those of
Euacanthe usambarensis generally have two guttules [
12
,
43
]. There are also more than 2.5%
nucleotide differences in LSU out of 925 base pairs among the strains of Yuxiensis granularis
(HKAS 109580) and Euacanthe usambarensis (GKM1221 and SMH4408), while more than
2.5% nucleotide differences in tef1 and rpb2 out of 731 and 1178 base pairs respectively
between the strains of Yuxiensis granularis (HKAS 109580) and Euacanthe usambarensis
(GKM1221).
Life 2021, 11, x FOR PEER REVIEW 13 of 19
Figure 3. Yuxiensis granularis (HKAS 109580, holotype). (a) Appearance of ascomata on twig. (b) Close-up of ascomata. (c)
Collabent ascoma. (d,e) Longitudinal sections of ascoma. (f) Peridium. (g) Quellkörper. (h) Munk pores (arrows). (i–k)
Asci. (l–p) Ascospores. Scale bars: (d,e) = 200 μm, (f) = 30 μm, (g) = 100 μm, (i–k) = 10 μm, (l–p) = 5 μm.
4. Discussion
Sordariomycetes is a frequently-studied class, with several taxa having been recently
introduced or revised [4,5,8,12,44]. The present study corroborates this fact, as supported,
firstly, by establishing a new saprobic genus, Yuxiensis, in Scortechiniaceae based on a dual
taxonomic approach. In addition to phylogeny, the familial placement of the new genus
within Scortechiniaceae is morphologically confirmed by the presence of the quellkörper in
Figure 3.
Yuxiensis granularis (HKAS 109580, holotype). (
a
) Appearance of ascomata on twig. (
b
) Close-up of ascomata.
(
c
) Collabent ascoma. (
d
,
e
) Longitudinal sections of ascoma. (
f
) Peridium. (
g
) Quellkörper. (
h
) Munk pores (arrows).
(i–k) Asci. (l–p) Ascospores. Scale bars: (d,e) = 200 µm, (f) = 30 µm, (g) = 100 µm, (i–k) = 10 µm, (l–p)=5µm.
4. Discussion
Sordariomycetes is a frequently-studied class, with several taxa having been recently
introduced or revised [
4
,
5
,
8
,
12
,
44
]. The present study corroborates this fact, as supported,
firstly, by establishing a new saprobic genus, Yuxiensis, in Scortechiniaceae based on a dual

Life 2021,11, 1011 13 of 17
taxonomic approach. In addition to phylogeny, the familial placement of the new genus
within Scortechiniaceae is morphologically confirmed by the presence of the quellkörper in
its centrum. Within Scortechiniaceae,Yuxiensis shares many overlapping characters with the
other genera. For instance, like almost all the other genera in the family with a known sexual
morph, Yuxiensis comprises ascomata which collapse upon drying, presence of munk pores
in the ascomatal wall and inconspicuous paraphyses [
4
,
8
]. It also has 8-spored asci, similar
to Biciliospora,Coronophorella,Euacanthe,Scortechinia, and Tympanopsis and long-pedicellate
asci like Biciliospora,Neofracchiaea,Scortechinia,Scortechiniella, and Scortechiniellopsis [
2
,
4
,
8
].
It is equally similar to most taxa of Neocryptosphaerella and Pseudocryptosphaerella in that
its ascomata are immersed to erumpent, appearing superficial when the substrate has
worn away [
2
,
12
]. The new genus however, demarcates itself from the other genera in the
family by several ways. Its ascomata are not seated on or surrounded by a well-developed,
conspicuous subiculum unlike many taxa of Biciliospora,Coronophorella,Euacanthe,Neofrac-
chiaea,Scortechinia,Scortechiniella,Scortechiniellopsis, or Tympanopsis [
2
,
4
,
8
]. The ascomata
of Yuxiensis are also devoid of a tomentose or setose surface as compared to Euacanthe,
Neofracchiaea, or some taxa of Neocryptosphaerella and Pseudocryptosphaerella [
2
,
8
]. Moreover,
the ascospores of Yuxiensis do not have conspicuous guttules unlike those of Euacanthe,
Neocryptosphaerella, and Pseudocryptosphaerella taxa and they lack appendage-like wall ex-
tensions on both ends, contrary to Biciliospora and Scortechiniella [
2
,
45
]. Since Yuxiensis is
phylogenetically closely related to Pseudocatenomycopsis and Euacanthe (Figure 1), more
details about their morpho-molecular comparisons have been given in the result parts
3.2.5 and 3.2.6 above. Based on all these morphological as well as phylogenetic differences,
Yuxiensis is herein introduced as a new genus.
The inclusion of Parasympodiellales in Coronophorales in the present study also points
toward the continuous amendment in the classification of Sordariomycetes. Herein, while
phylogeny supports the addition of Parasympodiellaceae to Coronophorales, this inclusion
is equally supported by the fact that taxa of Parasympodiellaceae have similar morpho-
logical characters with several taxa in other families of Coronophorales (Ceratostomataceae,
Chaetosphaerellaceae,Scortechiniaceae ) in terms of unbranched or branched, generally brown
and often septate conidiophores or conidia produced in chains [
4
,
8
,
11
]. Parasympodiellaceae
distinguishes itself from the other families mainly by its sympodial and unbranched coni-
diogenous cells which undergo schizolytic or rhexolytic secession to form arthroconidia.
Furthermore, the incertae sedis taxon Arthrocristula is synonymized under Parasympodiella,
with Arthrocristula hyphenata recombined to Parasympodiella hyphenata and representing
another synasexual morph of Parasympodiella. This arthrocristula-like synasexual morph of
Parasympodiella is typically characterized by branched conidiophores and conidiogenous
cells which secede rhexolytically to give rise to arthroconidia. It is also different from
the stylaspergillus-like synasexual morph of Parasympodiella which is generally character-
ized by pale brown, phialidic conidiogenous cells originating from terminal or intercalary
vesicle-like cells and filiform conidia which are produced in slimy masses [13].
An additional collection of Fracchiaea myricoides (HKAS 115760) was also made in
the present study, and sequence data for the same have been used in the phylogenetic
analyses and deposited in GenBank (Table 1). The latter species was initially introduced as
Coronophora myricoides based on LSU and ITS sequence data and the differences mentioned
between this taxon and the type species of Coronophora,C. gregaria was mainly based on the
shapes of the ascomata and ascospores [
46
]. Huang et al. [
12
] recently synonymized this
species to Fracchiaea myricoides; our collection supports this synonymy and the inclusion of
‘Coronophora myricoides’ in Fracchiaea (Nitschkiaceae) (Figure 1).
Despite the advancement towards a natural classification of Sordariomycetes, uncertain-
ties and confusions still prevail, as we note in the case of Parasympodiella longispora (CBS
544.84 and KACC 41225) (in Parasympodiellaceae clade, Figure 1). The latter is currently
known as ‘Bahusakala longispora’ in Index Fungorum and MycoBank, with Parasympodiella
longispora as an (obligate) synonym. The type strain of ‘Bahusakala longispora’, CBS 544.84,
sequenced by Vu et al. [
19
], clusters in the Parasympodiellaceae clade (Figure 1) with good

Life 2021,11, 1011 14 of 17
statistical support. Bahusakala longispora was introduced by Tokumasu and Tubaki [
47
] as a
taxon with conidiophores that are sympodial, rarely branched, erect in the lower part and
become geniculate (zig-zag, as mentioned in the original description) in the upper part.
Furthermore, the conidiogenous cells are hyaline, originating at regular intervals, while
the arthroconidia, subhyaline to pale yellow. Chlamydospores are produced in vegeta-
tive hyphae. However, the species was later synonymized to Parasympodiella longispora
since its morphology (based on its type) matched the description of Parasympodiella more
accurately [
48
]. Bahusakala taxa have been reported to have conidiophores that branch at
irregular intervals to produce brown conidiogenous hyphae at the terminal and intercalary
positions. The conidia are usually brown to dark brown and originate from random disar-
ticulation of the main conidiophore axes and conidiogenous hyphae [
42
,
48
–
50
]. Based on
the description of its type (and placement in the present phylogenetic tree), the species is
better accommodated in Parasympodiella than Bahusakala. We may as well adopt a broader
taxonomic perspective and decide that both Parasympodiella and Bahusakala are congeneric
since, despite their morphological differences, the two genera are also characterized by
similar features. Both are hyphomycetes with erect and septate conidiophores and produce
arthroconidia which secede schizolytically [
42
,
50
]. However, no molecular data for the
type species, B. olivaceonigra is yet available to enable any definite phylogenetic placement
and eventually to confirm a taxonomic conclusion for Bahusakala.
We also take note that the GenBank accession numbers of the sequence data for
Neocryptosphaerella globosa (GKM471N) that we use in our phylogeny have been assigned
under different strain numbers, namely, LSU (GenBank FJ968977: strain GKM469N), tef1
(GenBank FJ969036: strain GKM471N), and rpb2 (GenBank FJ968935: strain GKM469N).
In the original manuscript [
2
], these accession numbers are under the strain number
Neocryptosphaerella globosa GKM471N. Therefore, we followed the original paper.
This remarkable finding of a new genus in a rarely collected order indicates how little
we know of the fungal diversity of Yunnan and the broader region, including areas such as
Thailand and Laos [
51
], with recent studies showing large numbers of novel taxa being
discovered [
52
,
53
]. Further studies in other countries and habitats across this region will
surely result in the discovery of numerous other taxa in Parasympodiella,Yuxiensis, and
other poorly known taxa of Coronophorales [54].
Author Contributions:
Conceptualization, D.B., D.N.W. and K.D.H.; data curation, D.N.W.; formal
analysis, D.B., D.N.W., S.S.N.M. and S.-K.H.; funding acquisition, D.N.W. and K.D.H.; investigation,
D.B. and S.S.N.M.; methodology, D.B., D.N.W. and S.S.N.M.; resources, D.N.W., P.E.M. and K.D.H.;
supervision, K.D.H.; writing—original draft, D.B., D.N.W., S.S.N.M., D.J.B. and S.-K.H.; writing—
review & editing, S.L., P.E.M. and K.D.H. All authors have read and agreed to the published version
of the manuscript.
Funding:
Dhanushka N. Wanasinghe would like to thank CAS President’s International Fellowship
Initiative (PIFI) for funding his postdoctoral research (number 2021FYB0005) and the Postdoctoral
Fund from Human Resources and Social Security Bureau of Yunnan Province. Kevin D. Hyde
thanks the Thailand Research Fund, grant RDG6130001 entitled “Impact of climate change on fungal
diversity and biogeography in the Greater Mekong Sub-region”.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement:
The nucleotide sequences generated in the present study are deposited
in GenBank (Table 1). The final alignment and phylogenetic tree have been submitted to TreeBASE
(submission ID: 28713, http://www.treebase.org/, accessed on 30 August 2021). Specimen has been
deposited in the herbarium of Cryptogams Kunming Institute of Botany Academia Sinica (HKAS).
Acknowledgments:
Digvijayini Bundhun gratefully acknowledges Mae Fah Luang University, the
Mushroom Research Foundation and the Center of Excellence in Fungal Research, Thailand, for re-
search support. Ruvishika S. Jayawardena, Eleni Gentekaki, Rajesh Jeewon, Chitrabhanu S. Bhunjun,
and Vedprakash G. Hurdeal are thanked for their advice and support throughout this study. Kevin

Life 2021,11, 1011 15 of 17
D. Hyde thanks Chiang Mai University for the award of a Visiting Professorship. All authors thank
Shaun Pennycook and Paul Kirk for their valuable suggestions.
Conflicts of Interest: The authors declare no conflict of interest.
Appendix A
Life 2021, 11, x FOR PEER REVIEW 16 of 19
Conflicts of Interest: The authors declare no conflict of interest.
Appendix A
Figure A1. Phylogram inferred from the Bayesian analysis of LSU–ITS–tef1–rpb2 matrix for Coro-
nophorales. The tree is rooted with Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pandanicola
(MFLUCC 16-0316), and Stachybotrys microspora (KLM 3-2). BYPP (≥0.95 PP) supports are given
Figure A1.
Phylogram inferred from the Bayesian analysis of LSU–ITS–tef1–rpb2 matrix for
Coronophorales. The tree is rooted with Emericellopsis alkalina (CBS 127350), Pseudohyaloseta pan-
danicola (MFLUCC 16-0316), and Stachybotrys microspora (KLM 3-2). BYPP (
≥
0.95 PP) supports are
given above or below the branches. Type strains are in bold while novelty and the recombined taxon
are in blue.

Life 2021,11, 1011 16 of 17
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