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Phylogenetic placement of new species and
combinations of Neopodoconis within Torulaceae
Ling Qiu
Jiangxi Agricultural University
Ya-Fen Hu
Jiangxi Agricultural University
Jing-Wen Liu
Jiangxi Agricultural University
Ji-Wen Xia
Shandong Agricultural University
Rafael F. Castañeda-Ruíz
instituto de investigaciones fundamentales en agricultura tropical "Alejandro de Humboldt"
Zhao-Huan Xu
Jiangxi Agricultural University
Jian Ma ( majian821210@163.com )
Jiangxi Agricultural University https://orcid.org/0000-0001-9783-1860
Research Article
Keywords: Asexual Ascomycota, Hyphomycetes, Saprobic fungi, Taxonomy
Posted Date: June 21st, 2022
DOI: https://doi.org/10.21203/rs.3.rs-1755529/v1
License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full
License
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Abstract
Five new species of
Neopodoconis
,
N. jiangxiensis
,
N. meilingensis
,
N. obclavata
,
N. saprophyticus
and
N. sinensis
are described and illustrated from specimens collected on dead branches of unidentied plants in China.
Phylogenetic analysis of partial DNA sequences of nuclear ribosomal small subunit (SSU) and nuclear ribosomal
large subunit (LSU), using Maximum-Likelihood (ML) and Bayesian Inference (BI), supported the establishment of the
ve new species, and indicated a close relationship to Torulaceae in Pleosporales, Dothideomycetes.
Sporidesmioides
and
Rostriconidium
are synonymised with
Neopodoconis
based on morphological and molecular
phylogenetic data, and the new combinations
Neopodoconis aquaticum
,
N. cangshanense
,
N. pandanicola
and
N.
thailandica
are proposed. A synoptic table to
Neopodoconis
species are provided.
Introduction
The genus
Neopodoconis
Rifai was established by Rifai (2008) for two combinations,
N. ampullacea
(Petch) Rifai
and
N. megasperma
(Boedijn) Rifai, derived from
Exosporium
Link (Link 1809), and regarding
N. ampullacea
as type
species. The genus is a morphologically well-characterised by acropleurogenous, solitary, euseptate conidia seceding
schizolytically from polytretic, integrated, terminal and later intercalary, elongated sympodially, blackly cicatricated
conidiogenous cells on macronematous, mononematous, unbranched conidiophores. The conidia are obclavate to
broadly obclavate, smooth walled or verruculous, brown but much paler towards the apex, with protruding truncate
dark scar at the base and distinctly rostrate at the apex. To date, only
N. ampullacea
and
N. megasperma
are
reported in
Neopodoconis
, which were collected from dead stem or dried stick in Java, Ceylon, Ghana, and Sierra
Leone (Rifai 2008; Indexfungorum 2022). However, no DNA sequence exists for both species in GenBank, and so that
Neopodoconis
is even unclear of its taxonomic placement within
Ascomycota
(Wijayawardene et al. 2020).
Neopodoconis
was segregated from
Exosporium
by Rifai (2008), both genera show similar conidial ontogeny, and
the nature of conidial septation was regarded as the only character for separating
Neopodoconis
from
Exosporium
.
Such an approach was also used as the fundamental criterion in distinguishing a number of hyphomycete genera
(e.g. Subramanian 1992; Wu and Zhuang 2005; Seifert et al. 2011; Ma et al. 2016), but the phylogenetic signicance
of conidial septation in
Neopodoconis
-
Exosporium
taxonomy has not been undertaken by molecular methods.
Based on sequence data, two new genera
Sporidesmioides
Jun F. Li, Phook. & K.D. Hydeand and
Rostriconidium
Z.L.
Luo, K.D. Hyde & H.Y. Su were respectively introduced by Li et al. (2016) and Su et al. (2018), but both genera were
not compared morphologically with the closely related genus
Neopodoconis
, although no DNA sequence exists for
Neopodoconis
species to be used in their molecular phylogeny. Remarkably,
Sporidesmioides
and
Rostriconidium
exhibit the essential characteristics of
Neopodoconis
. These striking morphological consistency and the incomplete
taxon and sequence sampling in the phylogenies of Li et al. (2016) and Su et al. (2018) call for a critical treatment of
the status of
Sporidesmioides
and
Rostriconidium
as distinct genera. Thus,
Sporidesmioides
and
Rostriconidium
were proposed as synonyms of
Neopodoconis
based on their distinct morphological features and phylogenetic
analyses of combined SSU and LSU sequence data, and the new combinations
Neopodoconis aquaticum
,
N.
cangshanense
,
N. pandanicola
and
N. thailandica
are proposed.
Jiangxi Province is located in the southeast of China, and in the south bank of the middle and lower reaches of the
Yangtze River. It covers 166, 900 km2 and has a mountainous topography with remarkable subtropical monsoon
climate. Its annual mean temperature is 16.3–19.5 ℃, and the mean annual precipitation is 1341–1943 mm, so it is
particularly suitable for the growth of microscopic fungi in plant remains. During our mycological surveys in this
Province, ve specimens of
Neopodoconis
were collected on dead branches of unidentied plants. Based on
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morphological and phylogenetic analyses of combined SSU and LSU sequence data, they were described as new to
science in the present study.
Materials And Methods
Isolates and Morphological analysis
Samples of dead branches were collected from humid environments and river bank in the forest ecosystems of
Jiangxi Province, China, and returned to the laboratory in Ziploc™ bags. Samples were processed and examined
following the methods described in Ma et al. (2011). Fungi were mounted in a drop of lactic acid on microscope
slides, and examined and photographed with an Olympus microscope (model BX 53), with a 100 × (oil immersion)
objective at the same background and scale. Adobe Photoshop 7.0 was used for image processing to assemble
photographs into images. Single-spore isolations were made on potato dextrose agar (PDA) following Goh (1999). All
fungal strains were stored in 10% sterilised glycerine at 4°C for further studies. The studied specimens and cultures
were deposited in the Herbarium of Jiangxi Agricultural University, Plant Pathology, Nanchang, China (HJAUP).
Dna Extraction, Pcr Amplication And Sequencing
Genomic DNA was extracted from fungal mycelia grown on PDA, using the Solarbio Fungi Genomic DNA Extraction
Kit following the manufacturer’s protocol (Solarbio, China). DNA sequence data was obtained from small and large
subunits of the nuclear ribosomal RNA genes (SSU, LSU). Primer sets used for these genes were as follows: SSU:
18S-F/18S-R and LSU: 28S1-F/28S3-R (Xia et al. 2017). The nal volume of the PCR reaction was 25 µl, containing 1
µl of DNA template, 1 µl of each forward and reward primer, 12.5 µl of 2 × Power Taq PCR MasterMix and 9.5 µl of
ddH2O. The PCR thermal cycling conditions of SSU and LSU were initialized at 94°C for 3 min, followed by 35 cycles
of denaturation at 94°C for 30 s, annealing at 55°C for 50 s, elongation at 72°C for 1 min, a nal extension at 72°C for
10 min, and nally kept at 4°C. The PCR products were checked on 1% agarose gel electrophoresis stained with
ethidium bromide. Purication and DNA sequencing were carried out at Beijing Tsingke Biotechnology Co., Ltd.
China.
Sequence Alignment And Phylogenetic Analyses
Phylogenetic analyses were performed by concatenated SSU and LSU sequence data. Sequences generated from
this study were analyzed with other similar sequences obtained from GenBank, and those derived from recent studies
in Li et al. (2016), Su et al. (2018) and Shen et al. (2021). The newly generated sequences together with other
sequences obtained from GenBank and the recent studies (Table1) were initially aligned using MAFFTv.7 (Katoh and
Standley 2013) on the onlineserver (http://maffTh.cbrc.jp/alignment/server/), and optimized manually when needed.
Phylosuite software v1.2.1 (Zhang et al. 2020) was used to construct the phylogenetic tree based on SSU and LSU
sequence data. The concatenated aligned dataset was analyzed separately using Maximum likelihood (ML) and
Bayesian inference (BI). Maximum likelihood phylogenies were inferred by using IQ-TREE (Anisimova et al. 2011;
Nguyen et al. 2015) under Edge-linked partition model for 1000 standard bootstraps. The optimal ML tree search was
conducted with 1000 separate runs using the default algorithm of the program from a random starting tree for each
run. The nal tree was selected among suboptimal trees from each run by comparing the likelihood scores using the
K80 + G for SSU and TRN + I + G for LSU substitution model. Bayesian Inference phylogenies were inferred using
MrBayes 3.2.6 (Ronquist et al. 2012) under partition model (2 parallel runs, 2000000 generations), in which the initial
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25% of sampled data were discarded as burn-in. The best-t model was K80 + I + G for SSU and SYM + I + G for LSU.
The trees were viewed in FigTree v. 1.4.4 (http://tree.bio.ed.ac.uk/software/gtree) and the layout of the trees was
done in Adobe Illustrator CS v. 5. Newly generated sequences were deposited in GenBank.
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Table 1
Isolates and sequences used in this study and newly generated sequences are indicated in bold
Name Voucher
information GenBank accession
numbers References
LSU SSU
Arthopyrenia salicis
CBMAI 1330 JN903536 – Passarini et al. (2013)
Biatriospora mackinnonii
CBS 110022 GQ387614 GQ387553 Ahmed et al. (2014)
Biatriospora mackinnonii
CBS 674.75 GQ387613 GQ387552 Ahmed et al. (2014)
Biatriospora marina
CY 1228 GQ925848 GQ925835 Suetrong et al. (2009)
Byssosphaeria salebrosa
SMH 2387 GU385162 – Mugambi and Huhndorf
(2009)
Dendryphion europaeum
CPC 23231 KJ869202 – Crous et al. (2014)
Dendryphion europaeum
CPC 22943 KJ869203 – Crous et al. (2014)
Elongatopedicellata lignicola
MFLUCC 15–
0642 KX421368 KX421369 Ariyawansa et al. (2015)
Exosporium livistonae
CBS 131313 JQ044446 – Crous et al. (2011)
Exosporium livistonicola
MUCC 190 MF951161 – Videira et al. (2017)
Exosporium stylobatum
CBS 160.30 JQ044447 – Crous et al. (2011)
Herpotrichia macrotricha
GKM 196N GU385176 – Mugambi and Huhndorf
(2009)
Hysterium angustatum
CBS 236.34 FJ161180 GU397359 Boehm et al. (2009)
Melanomma pulvis-pyrius
CBS 124080 GU456323 GU456302 Zhang et al. (2009)
Monotosporella tuberculata
CBS 256.84 GU301851 – Schoch et al. (2009)
Neooccultibambusa
chiangraiensis
MFLUCC 12–
0584 KU764699 KU712458 Doilom et al. (2017)
Neopodoconis aquaticum
MFLUCC 16-1113 MG208143 – Su et al. (2018)
Neopodoconis aquaticum
KUMCC 15–0297 MG208144 – Su et al. (2018)
Neopodoconis cangshanense
MFLUCC 20–
0147 MW010285 – Shen et al. (2021)
Neopodoconis jiangxiensis HJAUP C0947 ON693846 ON693847 This Study
Neopodoconis meilingensis HJAUP C0905 ON693849 ON693843 This Study
Neopodoconis obclavata HJAUP C0829 ON693848 ON693844 This Study
Neopodoconis pandanicola
MFLUCC 20–
0145 MW010280 MW010282 Shen et al. (2021)
Neopodoconis pandanicola
KUMCC 17–0176 MH260318 MH260358 Tibpromma et al. (2018)
Neopodoconis saprophyticus HJAUP C0830 ON693851 ON705129 This Study
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Name Voucher
information GenBank accession
numbers References
LSU SSU
Neopodoconis sinensis HJAUP C0909 ON693845 ON693850 This Study
Neopodoconis thailandica
MFLUCC 13–
0840 KX437757 KX437759 Li et al. (2016)
Neopodoconis thailandica
KUMCC 16 − 0012 KX437758 KX437760 Li et al. (2016)
Neoroussoella bambusae
MFLUCC 11–
0124 KJ474839 – Liu et al. (2014)
Occultibambusa bambusae
MFLUCC 13–
0855 KU863112 KU872116 Dai et al. (2017)
Occultibambusa bambusae
MFLUCC 11–
0394 KU863113 KU872117 Dai et al. (2017)
Occultibambusa fusispora
MFLUCC 11–
0127 KU863114 – Dai et al. (2017)
Occultibambusa pustule
MFLUCC 11–
0502 KU863115 KU872118 Dai et al. (2017)
Paradictyoarthrinium
diffractum
MFLUCC 12–
0557 KP744497 – Liu et al. (2015)
Paradictyoarthrinium
diffractum
MFLUCC 13–
0466 KP744498 KP753960 Liu et al. (2015)
Paradictyoarthrinium
tectonicola
MFLUCC 12–
0556 KP744499 – Liu et al. (2015)
Paradictyoarthrinium
tectonicola
MFLUCC 13–
0465 KP744500 KP753961 Liu et al. (2015)
Pleomassaria siparia
CBS 279.74 DQ678078 DQ678027 Tanaka et al. (2010)
Prosthemium stellar
CBS 126964 AB553781 AB553650 Tanaka et al. (2010)
Roussoella angustior
MFLUCC 15–
0186 KT281979 – Ariyawansa et al. (2015)
Roussoella chiangraina
MFLUCC 10–
0556 KJ474840 – Liu et al. (2014)
Roussoella hysterioides
CBS 125434 AB524622 AB524481 Ahmed et al. (2014)
Roussoella intermedia
CBS 170.96 KF443382 KF443390 –
Roussoella magnatum
MFLUCC 15–
0185 KT281980 – Ariyawansa et al. (2015)
Roussoella nitidula
MFLUCC 11–
0182 KJ474843 – Liu et al. (2014)
Roussoella nitidula
MFLUCC 11–
0634 KJ474842 – Liu et al. (2014)
Roussoella scabrispora
MFLUCC 11–
0624 KJ474844 – Liu et al. (2014)
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Name Voucher
information GenBank accession
numbers References
LSU SSU
Roussoella siamensis
MFLUCC 11–
0149 KJ474845 KU872125 Liu et al. (2014)
Roussoella thailandica
MFLUCC 11–
0621 KJ474846 – Liu et al. (2014)
Roussoella pustulans
KT 1709 AB524623 AB524482 Tanaka et al. (2009)
Roussoellopsis macrospora
MFLUCC 12 −
0005 KJ474847 KJ739608 Liu et al. (2014)
Roussoellopsis tosaensis
KT 1659 AB524625 AB524484 Tanaka et al. (2009)
Seriascoma didymospora
MFLUCC 11–
0179 KU863116 KU872119 Dai et al. (2016)
Seriascoma didymospora
MFLUCC 11–
0194 KU863117 KU872120 Dai et al. (2017)
Sporidesmium australiense
HKUCC 10833 DQ408554 – Shenoy et al. (2006)
Torula herbarum
CBS 111855 KF443386 KF443391 Crous et al. (2015)
Torula herbarum
CBS 379.58 KF443383 KF443388 Crous et al. (2015)
Torula hollandica
CBS 220.69 KF443384 KF443389 Crous et al. (2015)
Versicolorisporium triseptatum
JCM 14775 AB330081 – Hatakeyama et al. (2008)
Results
Molecular Phylogeny
The phylogenetic analyses based on SSU and LSU sequences was used to infer the relationships of the new taxon
and its morphologically similar species (Fig.1). A combined dataset of 59 characters (SSU and LSU sequence data)
with 48 taxa analyzed either under ML or Bayesian criteria resulted in trees which were topologically congruent with
respect to the position of the new taxa investigated herein including
Hysterium angustatum
(CBS 236.34) as the
outgroup. Maximum likelihood and Bayesian Inference analyses of the combined dataset resulted in phylogenetic
reconstructions with largely similar topologies, and the best scoring RAxML tree is shown in Fig.1. Bootstrap support
values for maximum likelihood (MLBS) higher than 85% and Bayesian posterior probabilities (BPP) greater than 0.95
are given above the nodes. Most of the core genera of
Torulaceae
(Crous et al. 2015) and
Roussoellaceae
(Liu et al.
2014) in Pleosporales (Wijayawardene et al. 2014) are included in our phylogenetic analysis (Fig.1), and the two
families
Torulaceae
and
Roussoellaceae
are represented with well-supported clades. All species collected in this
study have a close phylogenetic relationship with
Sporidesmioides
and
Rostriconidium
, and well-supported in the
phylogenetic analyses as members of the Torulaceae. In the phylogenetic reconstructions based on analysis of the
combined SSU and LSU dataset, all the new taxa are well-separated with high bootstrap support as shown in Fig.1.
The newly collected
Neopodoconis meilingensis
and
N. saprophyticus
composed a clade is sister to the new
combinations
N. cangshanense
and
N. pandanicola
with high statistical support (MLBS = 96%, BPP = 1.00). The
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newly collected
N. obclavata
,
N. jiangxiensis
and
N. sinensis
formed a well-supported monophyletic clade, and was
placed sister to the new combination
N. thailandica
with strong support (MLBS = 98%, BPP = 1.00).
Taxonomy
Neopodoconis jiangxiensis Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, sp. nov. (Fig.2)
Index Fungorum number
: IF559728
Etymology
In reference to the province where the type specimen was found.
Holotypus
HJAUP M0947
Colonies on natural subatrate effuse, scattered, hairy, brown to black. Mycelium partly supercial, partly immersed in
the substratum, composed of branched, septate, pale brown to brown, smooth hyphae. Conidiophores
macronematous, mononematous, unbranched, erect, straight to curved, cylindrical, brown to pale brown, smooth,
thick-walled, septate, up to 185 µm long, 10–12 µm thick. Conidiogenous cells polytretic, integrated, terminal, later
becoming intercalary, cylindrical, brown, smooth, elongated sympodially, blackly cicatricated, with thickened and
blackened scars. Conidial secession schizolytic. Conidia solitary, acropleurogenous, simple, brown to pale brown,
obclavate, rostrate, sometimes smooth walled, but usually verruculose, 6–16-septate, with a thick, black truncate scar
at base and pale pigment cell above the scar, 80–170 µm long, 14–18 µm thick in the broadest part, tapering to 2.5–
5 µm near the apex.
Material examined China, Jiangxi Province, Nanchang, Meiling Scenic Spot, on dead branches of an unidentied
broadleaf tree, 10 July 2020, L. Qiu, HJAUP M0947 (Holotype), ex-type living culture HJAUP C0947.
Notes Phylogenetic analysis showed that our isolate clustered together and formed a sister clade with the isolate of
N. obclavata
, but is well-separated with strong statistical bootstrap value support (MLBS = 95%; BPP = 1.00).
Neopodoconis jiangxiensis
is morphologically distinguished from
N. obclavata
which has longer conidiophores (up
to 400 µm vs. up to 185 µm) and smooth conidia mostly with 8-septate.
Neopodoconis jiangxiensis
is also
morphologically most similar to
N. ampullacea
, but clearly differs in the size of conidiophores (up to 185 × 10–12
µm vs. up to 300 × 10–13 µm) and conidia (80–170 × 14–18 µm vs. 80–220 × 16–22), and in its conidia with fewer
septa (6–16-septate vs. 5–20-septate) and narrower apex (2.5–5 µm vs. 4–7 µm).
Neopodoconis meilingensis Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, sp. nov. (Fig.3)
Index Fungorum number
: IF559729
Etymology
In reference to the locality where the type specimen was found.
Holotypus
HJAUP M0905
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Colonies on natural subatrate effuse, scattered, hairy, brown to black. Mycelium partly supercial, partly immersed in
the substratum, composed of branched, septate, pale brown to brown, smooth hyphae. Conidiophores
macronematous, mononematous, unbranched, erect, straight to curved, cylindrical, brown to dark brown, smooth,
thick-walled, septate, up to 400 µm long, 7.5–10 µm thick. Conidiogenous cells polytretic, integrated, terminal, later
becoming intercalary, cylindrical, dark brown, smooth, elongated sympodially, blackly cicatricated, with thickened and
blackened scars. Conidial secession schizolytic. Conidia solitary, rarely catenate, acropleurogenous, simple,
obclavate, rostrate, brown, smooth, 7–20-septate, slightly constricted at some septa, with a thick, black truncate scar
at base and pale pigment cell above the scar, 90–230 µm long, 12–17 µm thick in the broadest part, tapering to 2.5–
6 µm near the apex.
Material examined China, Jiangxi Province, Nanchang, Meiling Scenic Spot, on dead branches of an unidentied
broadleaf tree, 10 July 2020, L. Qiu, HJAUP M0905 (Holotype), ex-type living culture HJAUP C0905.
Notes Phylogenetic analysis showed that our isolate clustered together and formed a sister clade with the isolate of
N. saprophyticus
, but is well-separated with strong statistical bootstrap value support (MLBS = 95%; BPP = 1.00).
Neopodoconis meilingensis
is morphologically distinguished from
N. saprophyticus
in its longer conidiophores (up to
400 × 7.5–10 µm vs. up to 325 × 8.5–10 µm) and solitary or rarely catenate, larger conidia (90–230 × 12–17 µm vs.
70–150 × 10–15 µm) with more septa (7–20-septate vs. 5–13-septate) and wider apex (2.5–6 µm vs. 1.5–2.5 µm).
Neopodoconis meilingensis
also differs from
N. ampullacea
in its longer and narrower conidiophores (up to 400 ×
7.5–10 µm vs. up to 300 × 10–13 µm) and solitary or rarely catenate, narrower conidia (90–230 × 12–17 µm vs. 80–
220 × 16–22 µm).
Neopodoconis obclavata Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, sp. nov. (Fig.4)
Index Fungorum number
: IF559730
Etymology
In reference to the obclavate conidia.
Holotypus
HJAUP M0829
Colonies on natural subatrate effuse, scattered, hairy, brown to black. Mycelium partly supercial, partly immersed in
the substratum, composed of branched, septate, pale brown to dark brown, smooth hyphae. Conidiophores
macronematous, mononematous, unbranched, erect, straight to curved, cylindrical, brown to dark black, paler
towards the apex, smooth, thick-walled, septate, up to 400 µm long, 10–12.5 µm thick. Conidiogenous cells polytretic,
integrated, terminal, later becoming intercalary, cylindrical, brown to dark brown, smooth, elongated sympodially,
blackly cicatricated, with thickened and blackened scars. Conidial secession schizolytic. Conidia solitary,
acropleurogenous, simple, obclavate, brown, rostrate, smooth, 7–11-septate (mostly 8), with a thick, black truncate
scar at base and pale pigment cell above the scar, 90–170 µm long, 10–20 µm thick in the broadest part, tapering to
3.5–6.5 µm near the apex.
Material examined China, Jiangxi Province, Yichun, Guanshan Mountain, on dead branches of an unidentied
broadleaf tree, 10 May 2020, L. Qiu, HJAUP M0829 (Holotype), ex-type living culture HJAUP C0829.
Page 10/21
Notes Our molecular data conrmed a clear separation with strong statistical support as shown in Fig.1.
Neopodoconis obclavata
shares similar characters with
N. ampullacea
in having macronematous, unbranched
conidiophores, and polytretic, integrated, terminal and intercalary, elongated sympodially, blackly cicatricated
conidiogenous cells with thickened and blackened scars and solitary, acropleurogenous, euseptate conidia. however,
N. obclavata
differs from
N. ampullacea
in the size of conidiophores (up to 400 × 10–12.5 µm vs. up to 300 × 10–13
µm) and conidia (90–170 × 15–18 µm vs. 80–220 × 16–22), and in its conidia usually with 8-euseptate.
Neopodoconis saprophyticus Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, sp. nov. (Fig.5)
Index Fungorum number
: IF559731
Etymology
In reference to the saprophytic habit on dead branches.
Holotypus
HJAUP M0830
Colonies on natural subatrate effuse, scattered, hairy, brown to black. Mycelium partly supercial, partly immersed in
the substratum, composed of branched, septate, pale brown to brown, smooth hyphae. Conidiophores
macronematous, mononematous, unbranched, erect, straight to curved, cylindrical, brown to dark brown, smooth,
thick-walled, septate, up to 325 µm long, 8.5–10 µm thick. Conidiogenous cells polytretic, integrated, terminal, later
becoming intercalary, cylindrical, dark brown to brown, smooth, elongated sympodially, blackly cicatricated, with
thickened and blackened scars. Conidial secession schizolytic. Conidia solitary, acropleurogenous, simple, brown to
pale brown, obclavate, rostrate, smooth, 5–13-septate (mostly 8), slightly constricted at some septa, with a thick,
black truncate scar at base and pale pigment cell above the scar, 70–150 µm long, 10–15 µm thick in the broadest
part, tapering to 1.5–2.5 µm near the apex.
Material examined China, Jiangxi Province, Yichun, Guanshan Mountain, on dead branches of an unidentied
broadleaf tree, 10 May 2020, L. Qiu, HJAUP M0830, (Holotype), ex-type living culture HJAUP C0830.
Notes
Neopodoconis saprophyticus
clusters with
N. meilingensis
, but is well-separated with high bootstrap support
as shown in Fig.1. Moreover,
N. saprophyticus
morphologically differs from
Neopodoconis meilingensis
in the size
of conidiophores (up to 325 × 8.5–10 µm vs. up to 400 × 7.5–10 µm) and conidia (70–150 × 10–15 µm vs. 90–230
× 12–17 µm), and in its conidia with fewer septa (5–13-septate vs. 7–20-septate) and narrower apex (1.5–2.5 µm vs.
2.5–6 µm).
Neopodoconis saprophyticus
also supercially resembles
N. ampullacea
, but differs in its narrower
conidiophores (8.5–10 µm vs. 10–13 µm wide) and smaller conidia (70–150 × 10–15 µm vs. 80–220 × 16–22 µm)
with fewer septa (5–13-septate vs. 5–20-septate) and narrower apex (1.5–2.5 µm vs. 4–7 µm).
Neopodoconis sinensis Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, sp. nov. (Fig.6)
Index Fungorum number
: IF559732
Etymology
In reference to the country in which the fungus was collected.
Holotypus
Page 11/21
HJAUP M0909
Colonies on natural subatrate effuse, scattered, hairy, brown to dark brown. Mycelium partly supercial, partly
immersed in the substratum, composed of branched, septate, smooth, pale brown to brown, smooth hyphae.
Conidiophores macronematous, mononematous, unbranched, erect, straight to curved, cylindrical, brown to dark
brown, slightly paler at the apex, smooth, thick-walled, 8–12-septate, up to 310 µm long, 8–10 µm thick.
Conidiogenous cells polytretic, integrated, terminal, later becoming intercalary, cylindrical, brown to dark brown,
smooth, elongated sympodially, blackly cicatricated, with thickened and blackened scars. Conidial secession
schizolytic. Conidia solitary, acropleurogenous, simple, obclavate, rostrate, smooth, 7–10-septate, slightly constricted
at some septa, brown to dark brown, with a thick, black truncate scar at base and pale pigment cell above the scar,
90–150 µm long, 13–15 µm thick in the broadest part, tapering to 3–5 µm near the apex.
Material examined China, Jiangxi Province, Nanchang, Meiling Scenic Spot, on dead branches of an unidentied
broadleaf tree, 10 July 2020, L. Qiu, HJAUP M0909 (Holotype), ex-type living culture HJAUP C0909.
Notes Our phylogenetic analyses showed that
N. sinensis
forms an independent clade (MLBS = 96%, BPP = 1.00), and
is phylogenetically related to
N. obclavata
and
N
.
jiangxiensis
.
Neopodoconis sinensis
morphologically shares
similarities with
N. ampullacea
, but is clearly different in the size of the conidiophores (up to 310×8–10 µm vs. up to
300 × 10–13 µm), conidia (90–150 × 13–15 µm vs. 80–220 × 16–22) and the number of conidial septa (7–10 vs.
5–20).
Neopodoconis sinensis
is also differs from
N. megasperma
which has shorter and wider conidia (60–90 ×
20–28.5 µm, 4–7-septate) with fewer eusepta.
New combinations from Rostriconidium and Sporidesmioides
Neopodoconis aquaticum (Z.L. Luo, K.D. Hyde & H.Y. Su) Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, comb. nov.
Index Fungorum number
: IF559733
Holotypus
MFLU 17–1415
Basionym
:
Rostriconidium aquaticum
Z.L. Luo, K.D. Hyde & H.Y. Su, Mycol. Progress 17(5): 536 (2018).
Notes For a detailed description of the species, see Su et al. (2018).
Rostriconidium aquaticum
is the generic type of
Rostriconidium
, which was established as a distinct genus based on a phylogenetic placement between
Sporidesmioides
and
Neotorula
. However, Su et al. (2018) didn't morphologically compare
Rostriconidium aquaticum
with the type species of
Neopodoconis
, although no DNA sequence exists for
Neopodoconis
species to be used in the
molecular phylogeny.
Rostriconidium aquaticum
exhibits the key characters of
Neopodoconis
, acropleurogenous,
solitary, euseptate conidia seceding schizolytically from polytretic, integrated, terminal and intercalary, elongated
sympodially, blackly cicatricated conidiogenous cells with thickened and blackened scars. Therefore,
R. aquaticum
is
clear congeneric with
Neopodoconis
, but not conspecic with the previously described species of
Neopodoconis
.
Rostriconidium aquaticum
morphologically shares similarities with
N. ampullacea
, but is clearly different in the size
of the conidiophores (370–590 × 13–17 µm vs. up to 300 × 10–13 µm), conidia (134–180 × 22–26 µm vs. 80–220 ×
16–22), and the number of conidial septa (8–9 vs. 5–20).
Neopodoconis cangshanense (H.W. Shen, Z.L. Luo & H.Y. Su) Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, comb. nov.
Page 12/21
Index Fungorum number
: IF559734
Holotypus
: MFLU: 20–0572
Basionym
:
Rostriconidium cangshanense
H.W. Shen, Z.L. Luo & H.Y. Su, Mycosystema 40(6): 1267 (2021).
Notes For a detailed description of the species, see Shen et al. (2021).
Neopodoconis cangshanense
was originally
assigned to
Rostriconidium
by Shen et al. (2021) based on the particular morphological characters and the support
of phylogeny. However, the genus
Rostriconidium
was proposed as a synonym of the earlier described
Neopodoconis
based on their distinct morphological features in the present study. Thus,
Rostriconidium cangshanense
is here
transferred to
Neopodoconis
, but differs from
N. ampullacea
which has smooth or verruculose, longer and narrower
conidia with more septa, and from
N. megasperma
which has shorter and wider conidia with the second cell from
below largest (Table2). In our phylogenetic analyses, the new combination,
Neopodoconis cangshanense
clusters
with
N. pandanicola
, but is well-separated with high bootstrap support.
Page 13/21
Table 2
Synoptic table for comparision of
Neopodoconis
species
Species Conidiophores Conidia References
Size (µm) Shape Size (µm
)Septation Verrucose Apical
width
(µm )
N. ampullacea
Up to 300 ×
10–13 Obclavate 80–
150(–
220) ×
16–22
5–20 Yes 4–7 Rifai
(2008)
N. aquaticum
370–590 ×
13–17 Fusiform to
pyriform 134–180
× 22–26 8–9 No – Su et al.
(2018)
N.
cangshanense
279–528 ×
12–14 Pyriform,
fusiform to
obclavate,
with a
sheath at
the tip
94–109 ×
21–24 6–8 No – Shen et al.
(2021)
N. jiangxiensis
Up to 185 ×
10–12 Obclavate 80–170 ×
14–18 6–16 Yes 2.5–5
N.
megasperma
Up to 480 ×
8–11.5 Broadly
obclavate,
occasionally
almost
turbinate or
subfusoidal
60–90 ×
20–28.5 4–7 No 3–4.5 Rifai
(2008)
N.
meilingensis
Up to 400 ×
7.5–10 Obclavate 90–230 ×
12–17 7–20 No 2.5–6 This study
N. obclavata
Up to 400 ×
10–12.5 Obclavate 90–170 ×
10–20 7–11
(mostly
8)
No 3.5–
6.5 This study
N.
pandanicola
360–485 ×
11–13 Obclavate,
with a
sheath at
the tip
55–110 ×
18–26 4–7 No – Tibpromma
et al.
(2018)
N.
saprophyticus
Up to 325 ×
8.5–10 Obclavate 70–150 ×
10–15 5–13
(mostly
8)
No 1.5–
2.5 This study
N. sinensis
Up to 310 ×
8–10 Obclavate 90–150 ×
13–15 7–10 No 3–5 This study
N. thailandica
(100–)120–
200 × 7–9 (−
9.5)
Ampulliform,
with a
sheath at
the tip
62.5–
80(− 97)
×
(16–)20–
25
6–7 Yes – Li et al.
(2016)
Neopodoconis pandanicola (Tibpromma & K.D. Hyde) Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, comb. nov.
Index Fungorum number
: IF559735
Holotypus
Page 14/21
HKAS 99620
Basionym
:
Rostriconidium pandanicola
Tibpromma & K.D. Hyde, Fungal Diversity 93:45 (2018).
Notes For a detailed description of the species, see Tibpromma et al. (2018). Tibpromma et al. (2018) assigned
R.
pandanicola
to the genus
Rostriconidium
based on morphological and molecular phylogenetic analyses.
Rostriconidium
is clear congeneric with
Neopodoconis
based on their distinct morphological features, and was
combined into
Neopodoconis
in the present study. The description and illustrations of
R. pandanicola
fully match
Neopodoconis
. It differs from
N. ampullacea
and
N. megasperma
in the size of the conidiophores and conidia, and
further from
N. ampullacea
in its smooth conidia with fewer septa (Table2), from
N. megasperma
which has conidia
with the second cell from below largest. Combined SSU and LSU phylogenetic analyses also conrm it as distinct
taxa.
Neopodoconis thailandica (Jun F. Li, Phook. & K.D. Hyde) Y.F. Hu, L. Qiu, R.F. Castañeda & Jian Ma, comb. nov.
Index Fungorum number
: IF559736
Holotypus
MFLU14-0827
Basionym
:
Sporidesmioides thailandica
Jun F. Li, Phook. & K.D. Hyde, Mycol. Progress 15(10): 1171 (2016).
Notes For a detailed description of the species, see Li et al. (2016).
Sporidesmioides thailandica
is the generic type of
Sporidesmioides
, which was established as a distinct monotypic genus based on a phylogenetic placement of
forming a distinct, monotypic clade clustering with Torulaceae. The morphological features of this fungus fully
match the existing genus
Neopodoconis
, but Li et al. (2016) didn't morphologically compare it with
Neopodoconis
species. Although no sequence data of
Neopodoconis
species are available, there is no doubt that the species
belongs to
Neopodoconis
. Therefore,
Sporidesmioides thailandica
is here transferred to
Neopodoconis
. It differs from
N. ampullacea
and
N. megasperma
in the size of the conidiophores and conidia, and further from them by its
conidial apex with a ap-like, hyaline sheath (Table2). Our molecular data also conrmed a clear separation with
strong statistical support.
Discussion
Rifai (2008) proposed the genus
Neopodoconis
to accommodate
Exosporium-
like species that have solitary or rarely
catenate, euseptate conidia, and so far only two species have been reported (Rifai 2008).
Neopodoconis
and its
closely related genus
Exosporium
show similar conidial ontogeny, and conidial euseptation was regarded as the only
character for separating
Neopodoconis
from
Exosporium
, and their phylogenetic placement is well-separated as
shown in Fig.1.
Sporidesmioides
and
Rostriconidium
are two aquatic genera within the family Torulaceae (Pleosporales,
Dothideomycetes) (Li et al. 2016; Su et al. 2018). Both genera are clear congeneric with
Neopodoconis
based on their
distinct morphological features, and were here synonymised with
Neopodoconis
under the current code (Turland & al.
2018: Art. 11.3). Our phylogenetic analyses of SSU and LSU sequence data also resolved a relationship of
Sporidesmioides
and
Rostriconidium
with
Neopodoconis
. The results show that the genus
Sporidesmioides
is closely
related to
N. jiangxiensis
,
N. obclavata
,
N. sinensis
, and formed a distinct phylogenetic clade with high bootstrap
values (MLBS = 98%; BPP = 1.00). The genus
Rostriconidium
is closely related to
N. saprophyticus
and
N. sinensis
,
Page 15/21
and formed a distinct clade with strong support (MLBS = 97%; BPP = 1.00). All the taxa are well-separated with high
bootstrap support as shown in Fig.1. Based on morphology and sequence data, we introduced ve new species,
Neopodoconis jiangxiensis
,
N. meilingensis
,
N. obclavata
,
N. saprophyticus
and
N. sinensis
, and four new
combinations,
N. aquaticum
,
N. cangshanense
,
N. pandanicola
and
N. thailandica
, and treated
Neopodoconis
in
Torulaceae, Pleosporales, Dothideomycetes. A synopsis of the morphological characters of these currently accepted
Neopodoconis
species is presented in Table 2.
Declarations
Acknowledgments This project was supported by the National Natural Science Foundation of China (Nos. 31970018,
32160006, 31360011).
Competing Interests The authors declare no competing interests.
Author contribution Conceived and designed the experiments: J Ma. Performed the experiments: L Qiu, YF HU, JW Liu
and ZH Xu. Analyzed the data: L Qiu and YF Hu. Wrote the paper: L Qiu, JW Xia, RF. Castañeda-Ruíz and J Ma. All
authors read and approved the nal manuscript.
Data availability The data, including the sequences on GenBank and specimen data on Index Fungorum will be
available to any researcher wishing to use them for non-commercial purposes, without breaching participant
condentiality.
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Figures
Figure 1
Page 17/21
Phylogenetic tree inferred from maximum likelihood and Bayesian inference analysis based on a concatenated
alignment of SSU and LSU sequences. Bootstrap support values for maximum likelihood (MLBS) equal to or greater
than 85% and Bayesian posterior probabilities (BPP) equal to or greater than 0.95 are shown above the nodes at the
rst and second position, respectively. Ex-type isolates are in red, and new combinations are indicated in bold. The
tree is rooted to
Hysterium angustatum
(CBS 236.34).
Figure 2
Neopodoconis jiangxiensis (HJAUP M0947). a Colony on PDA (surface and reverse). b, c Conidiophores,
conidiogenous cells, and conidia. d, e Conidia.
Page 18/21
Figure 3
Neopodoconis meilingensis (HJAUP M0905). a Colony on PDA (surface and reverse). b Conidiophores,
conidiogenous cells, and conidia. c Conidiophores and conidiogenous cells. d, e Conidia.
Page 19/21
Figure 4
Neopodoconis obclavata (HJAUP M0829). a Colony on PDA (surface and reverse). b Conidiophores, conidiogenous
cells, and conidia. c Conidiophores and conidiogenous cells. d Conidiogenous cells and conidia. e, f Conidia.
Page 20/21
Figure 5
Neopodoconis saprophyticus (HJAUP M0830). a Colony on PDA (surface and reverse). b Conidiophores,
conidiogenous cells, and conidia. c Conidiophores and conidiogenous cells. d Conidiogenous cells and conidia. e, f
Conidia.
Page 21/21
Figure 6
Neopodoconis sinensis (HJAUP M0909). a Colony on PDA (surface and reverse). b, c Conidiophores, conidiogenous
cells, and conidia. d Conidia.
