Kirschsteiniothelia thailandica sp. nov. (Kirschsteiniotheliaceae) from Thailand

Abstract

A Kirschsteiniothelia species was found on decayed twigs of Ficus microcarpa collected at the Garden of Medicinal Plants in Chiang Rai, Thailand. Phylogenetic analyses of combined ITS, LSU and SSU sequence data showed the new isolate was phylogenetically distinct from other known species of Kirschsteiniothelia. Accordingly, Kirschsteiniothelia thailandica sp. nov. is described and illustrated and compared with other species in the genus.

Full text

Phytotaxa 490 (2): 172–182
https://www.mapress.com/j/pt/
Copyright © 2021 Magnolia Press Article PHYTOTAXA
ISSN 1179-3155 (print edition)
ISSN 1179-3163 (online edition)
172 Accepted by Sajeewa Maharachchikumbura: 4 Jan. 2021; published: 12 Mar. 2021
https://doi.org/10.11646/phytotaxa.490.2.3
Kirschsteiniothelia thailandica sp. nov. (Kirschsteiniotheliaceae) from Thailand
YA-RU SUN1,2,3,5, RUVISHIKA S. JAYAWARDENA2,3,6, KEVIN D. HYDE2,3,4,7 & YONG WANG1,8,*
1 Department of Plant Pathology, College of Agriculture, Guizhou University, Guiyang, Guizhou 550025, China.
2 Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand.
3 School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand.
4 Institute of Plant Health, Zhongkai University of Agriculture and Engineering, Haizhu District, Guangzhou 510000, China.
5
�
yarusun5@gmail.com; https://orcid.org/0000-0001-5549-102
6
�
ruvi.jaya@yahoo.com; https://orcid.org/0000-0001-7702-4885
7
�
kdhyde3@gmail.com; https://orcid.org/0000-0002-2191-0762
8
�
yongwangbis@aliyun.com; https://orcid.org/0000-0003-3831-2117
*Corresponding author: YONG WANG,
�
yongwangbis@aliyun.com
Abstract
A Kirschsteiniothelia species was found on decayed twigs of Ficus microcarpa collected at the Garden of Medicinal Plants
in Chiang Rai, Thailand. Phylogenetic analyses of combined ITS, LSU and SSU sequence data showed the new isolate was
phylogenetically distinct from other known species of Kirschsteiniothelia. Accordingly, Kirschsteiniothelia thailandica sp.
nov. is described and illustrated and compared with other species in the genus.
Keywords: 1 new taxon, hyphomycetes, Kirschsteiniotheliales, multi-gene phylogeny, taxonomy
Introduction
Kirschsteiniothelia was introduced by Hawksworth (1985) based on the type species Kirschsleiniothelia aethiops.
It has both asexual and sexual morphs. The sexual morph of Kirschsleiniothelia has brown or black, hemisphaerical
or subglobose ascomata, cylindrical-clavate, bitunicate, 8-spored asci and ellipsoidal, brown to dark brown, smooth-
walled, 1–2-septate ascospores (Hawksworth 1985, Boonmee et al. 2012, Hyde et al. 2013, Mehrabi et al. 2017).
There are two types of asexual morphs in Kirschsteiniothelia, viz. dendryphiopsis-like and sporidesmium-like.
Dendryphiopsis was linked with Kirschsteiniothelia, which was confirmed by Hawksworth (1985) and Boonmee et
al. (2012). Subsequently, Su et al. (2016) reported a sporidesmium-like asexual morph (Kirchsteiniothelia submerse)
in Kirschsteiniothelia. Wijayawardene et al. (2014) recommended using Kirschsteiniothelia over Dendryphiopsis
considering fewer names changes.
Kirschsteiniothelia was assigned to Pleosporaceae by Hawksworth (1985) and Barr (1987). Barr (1993) suggested
that Kirschsteiniothelia should be transferred to the Pleomassariaceae considering its asexual morph, host and
morphology. Schoch et al. (2006) proposed that Kirschsteiniothelia should has its own family as the type species K.
aethiops did not group with Pleosporaceae in their phylogenetic analyses. In a further study, Schoch et al. (2009) showed
that K. maritima clustered within Mytilinidion (Mytilinidiaceae), while K. elaterascus grouped within Morosphaeria
(Morosphaeriaceae) (Suetrong et al. 2009). Kirschsteiniotheliaceae was established to accommodate Kirschsteiniothelia
based on morphology and phylogenetic analyses (Boonmee et al. 2012). In this study, Halokirschsteiniothelia was
introduced to accommodate formerly species K. maritima, and K. elaterascus was transferred to Morosphaeria
(Boonmee et al. 2012). Kirschsteiniotheliaceae was distantly related to other orders in Dothideomycetes in the study
performed by Hernandez-Restrepo et al. (2017). Therefore, a new order Kirschsteiniotheliales was established.
Hongsanan et al. (2020) provided a refined, updated document on families of Dothideomycetes. Kirschsteiniotheliales
is a monotypic order grouped with Asterinales in Dothideomycetidae, but the monophyly of these two orders is not
well-supported. Moreover, Kirschsteiniotheliales and Asterinales diverged approximately 221 MYA in divergence
time estimates (Hongsanan et al. 2020).

KIRSCHSTEINIOTHELIA THAILANDICA SP. NOV. Phytotaxa 490 (2) © 2021 Magnolia Press • 173
We are studying the fungi growing on medicinal plants in China and Thailand to record diseases and other
beneficial properties (Long et al. 2019, Sun et al. 2020, Zhang et al. 2020). A specimen of Kirschsteiniothelia was
collected at the Garden of Medicinal Plants in Mae Fah Luang University, Chiang Rai, Thailand. Morphological study
and molecular analysis showed that our isolate represents a phylogenetically distinct species of Kirschsteiniothelia.
Therefore, we introduce K. thailandica as a novel species.
Materials & methods
Collections and examination of specimens
Fresh samples of decayed wood were collected from a garden of medicinal plants, Mae Fah Luang University, Chiang
Rai, Thailand (November 2019). We used a stereomicroscope (SteREO Discovery. V12, Carl Zeiss Microscopy
GmBH, Germany) to observe fungal structures on natural substrate. The fungal structures were transferred to a small
drop of water on a slide using a syringe needle and covered with a cover glass. The fungi were checked by a Nikon
ECLIPSE Ni compound microscope (Nikon, Japan) and photographed with a Nikon DS-Ri2 digital camera (Nikon,
Japan). Images used for figures were processed with Adobe Photoshop CS6 software (Adobe Systems, USA) and
measurements were made with the Tarosoft (R) Image Frame Work software.
The single-spore isolation was made on potato dextrose agar (PDA). Germinated conidium was transferred to new
PDA plates and incubated at 26 ºC for four weeks following the method of Chomnunti et al. (2014). Herbaria materials
were deposited in the Fungarium of Mae Fah Luang University (MFLU), Chiang Rai, Thailand. Pure cultures and
tubes were deposited in the Mae Fah Luang University Culture Collection (MFLUCC). Faces of fungi (FoF) and Index
Fungorum numbers were acquired as described in Jayasiri et al. (2015) and Index Fungorum (2021).
DNA extraction, PCR amplification and sequencing
Genomic DNA was extracted from scraped fresh fungal mycelium grown on PDA using the Genomic DNA Extraction
Kit (GD2416) following the manufacture’s protocol. DNA amplification was performed as the method described by
Wijesinghe et al. (2020). PCR was carried out in 20 μl reaction volume which contained 10 μl 2x PCR Master Mix, 1 μl
of each primer, 1 μl template DNA and 7μl ddH2O. Primer pair ITS5/ITS4 were used to amplify the internal transcribed
spacer regions 1 and 2 including the intervening 5.8S nuclear ribosomal DNA (ITS) region and NS1/NS4 for the
nuclear 18S ribosomal DNA (SSU) (White et al. 1990) . Primers LR0R and LR5 were used to amplify the nuclear 28S
ribosomal DNA (LSU) region (Vilgalys & Hester 1990). The PCR thermal cycle program for ITS, LSU and SSU were
as follows: 3 min at 94 ºC (initial denaturation), followed by 35 cycles of 30 s at 94 ºC (denaturation), 30 s at 52 ºC
(annealing), 1 min at 72 ºC (extension), with a final extension of 8 min at 72 ºC.
Phylogenetic analyses
The sequences (Table 1) used for phylogenetic analysis were downloaded from GenBank (https://www.ncbi.nlm.nih.
gov/genbank/) according to a BLASTn searching in the sequences from type material and relevant literature (Boonmee
et al. 2012, Mehrabi et al. 2017, Bao et al. 2018, Jiang et al. 2020). Alignments of single gene were performed with
MAFFT v7.2.1.2 (Katoh & Standley 2013). AliView (Larsson 2014) was used to check the alignments and change the
format. Ambiguous regions of the alignments were trimmed by TrimAl v 1.2 with gappyout option (Capella-Gutiérrez
et al. 2009). Sequence Matrix was used for combining single gene alignments in a concatenated alignment (Vaidya et
al. 2011). The newly generated sequences were submitted to GenBank.
The maximum likelihood (ML) analysis was carried out with IQ-tree (Nguyen et al. 2015, Chernomor et al. 2016).
The best fitting substitution models were chosen by jModelTest2 (Darriba et al. 2012) under the Akaike Information
Criterion (AIC) algorithm for each dataset. The TPM2uf + I + G model was selected for ITS dataset, and TrNef + I +
G for LSU, and TrNef + I + G for SSU. Nonparametric bootstrap iterations were run with 1,000 replicates.
The maximum parsimony (MP) analysis was performed with PAUP v. 4.0b10 (Swofford 2002). Gaps were treated
as missing data and every character was unordered with equal weight. Bootstrap (BS) analysis was used to estimate
clade stability, including 1,000 replicates, each with 10 replicates of random stepwise addition of taxa (Hillis & Bull
1993).

SUN ET AL.
174 • Phytotaxa 490 (2) © 2021 Magnolia Press
Bayesian inference (BI) analysis was conducted in MrBayes 3.2 (Ronquist et al. 2012). MrModeltest v.2.3
(Nylander 2004) was used to estimate the best evolutionary models under the Akaike Information Criterion (AIC).
GTR + I + G model was chosen for each locus. Six simultaneous Markov chains were run for 1,000,000 generations
and trees were sampled every 100 generations. The first 25% trees were discarded and the remaining trees were used
to calculate Bayesian posterior probabilities (BPP) in the majority rule consensus tree.
Phylogenetic trees were viewed with FigTree v1.4.4 (Rambaut 2009) and modified in Adobe Illustrator CS6 software
(Adobe Systems, USA).
TABLE 1. Isolates and sequences used for molecular analysis. The newly generated sequence is indicated in red. Ex-
type strains are in bold.
Taxon strain ITS LSU SSU
Acrospermum adeanum M133 EU940180 EU940104 EU940031
Acrospermum compressum M151 EU940161 EU940084 EU940012
Acrospermum gramineum M152 EU940162 EU940085 EU940013
Anisomeridium ubianum MPN94 - GU327709 JN887379
Flavobathelium epiphyllum MPN67 - GU327717 JN887382
Kirschsteiniothelia aethiops CBS 109.53 - AY016361 AY016344
Kirschsteiniothelia aethiops MFLUCC 16–1104 MH182583 MH182589 MH182615
Kirschsteiniothelia aethiops S–783 MH182586 MH182595 MH182617
Kirschsteiniothelia aethiops MFLUCC 15–0424 KU500571 KU500578 KU500585
Kirschsteiniothelia aquatica MFLUCC 17–1685 MH182587 MH182594 MH182618
Kirschsteiniothelia arasbaranica IRAN 2509C KX621986 KX621987 KX621988
Kirschsteiniothelia arasbaranica IRAN 2508C KX621983 KX621984 KX621985
Kirschsteiniothelia cangshanensis MFLUCC 16–1350 MH182584 MH182592 -
Kirschsteiniothelia fluminicola MFLUCC 16–1263 MH182582 MH182588 -
Kirschsteiniothelia lignicola MFLUCC 10–0036 HQ441567 HQ441568 HQ441569
Kirschsteiniothelia phoenicis MFLUCC 18–0216 MG859978 MG860484 MG859979
Kirschsteiniothelia rostrata MFLUCC 15–0619 KY697280 KY697276 KY697278
Kirschsteiniothelia rostrata MFLUCC 16–1124 - MH182590 -
Kirschsteiniothelia submersa MFLUCC 15–0427 KU500570 KU500577 KU500584
Kirschsteiniothelia submersa S–481 - MH182591 MH182616
Kirschsteiniothelia submersa S–601 MH182585 MH182593 -
Kirschsteiniothelia tectonae MFLUCC 12–0050 KU144916 KU764707 -
Kirschsteiniothelia thailandica MFLUCC 20–0116 MT985633 MT984443 MT984280
Kirschsteiniothelia thujina JF 13210 KM982716 KM982718 KM982717
Megalotremis verrucosa MPN104 - GU327718 JN887383
Phyllobathelium anomalum MPN 242 - GU327722 JN887386
Phyllobathelium firmum ERP 3175 - GU327723 -
Pseudorobillarda eucalypti MFLUCC 12–0422 KF827451 KF827457 KF827463
Pseudorobillarda phragmitis CBS 398.61 MH858101 EU754203 EU754104
Strigula guangxiensis HMAS-L0138040 NR146255 MK206256 -
Strigula macrocarpa HMAS-L0141394 - MK206240 MK206221
Strigula nemathora MPN 72 - JN887405 JN887389
Strigula nitidula HMAS-L0139358 - MN788374 MN788375
Strigula sinoaustralis HMAS-L0137204 - MK206249 -
Strigula univelbiserialis HMAS-L0137657 - MK206243 MK206224
Tenuitholiascus porinoides HMAS-L0139638 -MK206259 MK352441
Tenuitholiascus porinoides HMAS-L0139639 - MK206258 MK352442
Tenuitholiascus porinoides HMAS-L0139640 - MK206260 MK352443

KIRSCHSTEINIOTHELIA THAILANDICA SP. NOV. Phytotaxa 490 (2) © 2021 Magnolia Press • 175
Results
Phylogenetic analyses
To better infer the phylogenetic position of the new taxon in Kirschsteiniothelia, nucleotide sequences of the ITS, LSU
and SSU were used. Thirty-six strains, representing four orders (Acrospermales, Kirschsteiniotheliales, Monoblastiales
and Strigulales), with 2 outgroup Pseudorobillarda eucalypti (MFLUCC 12–0422) and P. phragmitis (CBS 398.61)
were analyzed. Maximum likelihood phylogenetic tree generated by IQ Tree with the best scoring (likelihood value =
-12960.346) was showed (Fig 1). The final combined dataset consisted of 2,146 characters (LSU: 742 bp, ITS: 392 bp,
SSU: 1012 bp), including alignment gaps. Among them, 1,368 characters were constant and 778 variable (including
575 parsimony-informative characters and 203 parsimony-uninformative). Fourteen equally most parsimonious trees
(Tree length = 2,178, CI = 0.548, RI = 0.707, RC = 0.388, HI = 0.452) were produced from the heuristic search. All
phylogenetic trees (ML, MP and BI) were similar in topology.
FIGURE 1. Maximum likelihood phylogenetic tree generated by IQ Tree, based on combined ITS, LSU and SSU sequence data. The tree
is rooted with Pseudorobillarda eucalypti (MFLUCC 12-0422) and P. phragmitis (CBS 398.61). The ex-type strains are indicated in bold
and the new isolates are in red and bold.

SUN ET AL.
176 • Phytotaxa 490 (2) © 2021 Magnolia Press
The phylogenetic analyses showed that the monophyly of Kirschsteiniotheliales was strongly supported by ML, MP
and BI analyses. Acrospermales, Monoblastiales and Strigulales clustered together and sister to Kirschsteiniotheliales
with strong support values (100% MLBS, 100% MPBS and 1.00 BPP). Kirschsteiniothelia thailandica formed a
distinct lineage, closely related to K. rostrata, K. tectonae and K. thujina (97% MLBS, 89% MPBS and 1.00 BPP).
Taxonomy
Kirschsteiniothelia thailandica Y.R. Sun, Yong Wang bis & K.D. Hyde, sp. nov.
Index Fungorum number: IF557949; Facesoffungi number: FoF 09289
Etymology: Referring the fungus collected from Thailand
Holotype: MFLU 20–0263
Saprobic on dead wood. Sexual morph: Undetermined. Asexual morph (Fig 2): Hyphomycetous. Colonies scattered,
effuse, brown to dark-brown, hairy on natural substrate. Mycelium composed of branched, smooth, septate, hyaline
hyphae. Conidiophores macronematous, mononematous, solitary, erect, cylindrical, straight or slightly flexuous,
unbranched, septate, brown to dark brown 55–93 × 7–10 μm ( = 75.5 × 9 μm, n = 15). Conidiogenous cells
monoblastic, integrated, terminal, cylindrical, brown to dark brown 9.5–21 × 7–10 μm ( = 16.5 × 8 μm, n = 15).
Conidia acrogenous, straight, solitary, obclavate, smooth-walled, 6–8-distoseptate, shiny, olivaceous or brown, hyaline
at apex, truncate at base, tapering towards apex, 74–110 μm long ( = 91 μm, n = 20), 13–20 μm at the widest ( = 17
μm, n = 20), with a conspicuous, gelatinous, hyaline sheath around tip.
Culture characteristics: Conidia germinated on PDA within 24 hours from single-spore isolation. Both ends
produced germ tubes. Colony diameter reached 25 mm after one month at indoor temperature on PDA media, circular,
surface rough, flat, dark-olivaceous from above, dark from below.
Material examined: THAILAND, Chiang Rai Province, Mae Fah Luang University, Garden of Medicinal Plants,
on twigs of Ficus microcarpa, 11 November 2019, Y.R. Sun, B4 (MFLU 20–0263, holotype; ex-type living culture
MFLUCC 20–0116).
Notes: We delete Kirschsteiniothelia emarceis because it’s a long branch in phylogenetic trees. Phylogenetic
analysis showed that K. thailandica clustered with K. rostrata, K. tectonae and K. thujina, and they formed a sister clade
with K. submersa. Kirschsteiniothelia thujina is known only its sexual morph (Hawksworth 1985). Kirschsteiniothelia
thailandica resembles to K. rostrate and K. tectonae in having unbranched, cylindrical conidiophores, integrated,
terminal conidiogenous cells and obclavate, rostrate conidia with mucilaginous sheaths. However, K. thailandica has
shorter conidiophores than K. rostrata (55–93 μm vs 190–450 μm). Kirschsteiniothelia thailandica differs from K.
tectonae in having shorter conidia (74–110 μm vs 135–150 μm) and shorter conidiophores (55–93 μm vs up to 200
μm). In addition, polymorphic nucleotides from the ITS sequence data showed the more than one fifth of bases were
different, and we also compared with other gene sequences (Table 2).
TABLE 2. Number of polymorphic nucleotide differences between K. thailandica compared with K. tectonae and K.
rostrata.
Taxon Isolate ITS LSU SSU
K. tectonae MFLUCC 12–0050 108 31 not available
K. rostrata MFLUCC 15–0619 98 37 38
K. rostrata MFLUCC 16–1124 not available 36 not available

KIRSCHSTEINIOTHELIA THAILANDICA SP. NOV. Phytotaxa 490 (2) © 2021 Magnolia Press • 177
FIGURE 2. Kirschsteiniothelia thailandica (MFLU 20–0263, holotype) a Colonies on natural substrate. b–d Conidiophores with conidia.
e Conidiogenous cell with conidium. f Conidiogenous cell. g–j Conidia. k Bipolar-germinated conidium. l, m Colony on PDA. Scale bars:
a = 100 μm, b–d = 50 μm, e–k = 20 μm.

SUN ET AL.
178 • Phytotaxa 490 (2) © 2021 Magnolia Press
Discussion
Kirschsteiniothelia has a worldwide distribution and most species have been found in tropical (Mehrabi et al. 2017), or
subtropical regions (Su et al. 2016, Bao et al. 2018). Most Kirschsteiniothelia species occur in terrestrial habitats, while
some occur in freshwater, such as Kirschsteiniothelia aquatica, K. cangshanensis, and K. fluminicola Z (Boonmee et
al. 2012, Li et al. 2016, Su et al. 2016, Hyde et al. 2017, Mehrabi et al. 2017, Bao et al. 2018) (Table 3).
Thirty-five species are accepted in Kirschsteiniothelia and six species are presently known from Thailand (Table
3). However with comprehensive collection, it is expected that many more taxa will be discovered as Thailand’s has
a high fungal diversity (Hyde et al. 2018) and the genus is not well-studied (Hyde et al. 2020). Two asexual morph
types are known in Kirschsteiniothelia (Boonmee et al. 2012, Su et al. 2016, Hyde et al. 2017, Bao et al. 2018).
Dendryphiopsis-like asexual morph has branched conidiophores, cylindrical, 3–4-septate, brown conidia with rounded
apices and an attenuated, truncate base, such as K. aethiops and K. goaensis (Pratibha et al. 2010, Su et al. 2016).
Sporidesmium-like asexual morph are characterized by unbranched conidiophores, oblong to obclavate, 6–27-septate,
olivaceous or brown conidia with or without sheath around tip, for example, K. aquatica, K. tectonae, and K. rostrata
(Li et al. 2016, Hyde et al. 2017, Bao et al. 2018). Moreover, it was noteworthy that K. binsarensis is the only species
with a dendryphiopsis-like asexual morph with obclavate conidia.
Kirschsteiniothelia thailandica was found on terrestrial dead wood. Phylogenetic analyses shows that it forms a
distinct lineage. Kirschsteiniothelia thailandica resembled with other species in Kirschsteiniothelia but it has a unique
dimensions.
TABLE 3. Summary of distribution, habitat, host and morphology type of Kirschsteiniothelia species.
Taxon Country Habitat Host Sexual morph Asexual morph
Kirschsteiniothelia abietina U.S.A Terrestrial Tsuga canadensis Determined Undetermined
K. acerina U.S.A Terrestrial Acer saccharum Determined Undetermined
K. aethiops
Belgium/
Germany/
U.S.A/China
Terrestrial/
Freshwater
Querus/carpinus/carpinus/
dead wood/submerged wood Determined Dendryphiopsis-like
K. aquatica China Freshwater Submerged wood Undetermined Sporidesmium-like
K. arasbaranica Iran Terrestrial Quercus petraea Determined Undetermined
K.arbuscula Unknown Unknown Unknown Undetermined Dendryphiopsis-like
K. atkinsonii Hawaii Terrestrial Freyeinetia arnoltii Determined Undetermined
K. atra U.S.A Terrestrial Dead wood Undetermined Dendryphiopsis-like
K. binsarensis India Terrestrial Dead twigs Undetermined Dendryphiopsis-like
K. biseptata Sri Lanka Terrestrial Dead wood Undetermined Dendryphiopsis-like
K. cangshanensis China Freshwater Submerged wood Undetermined Sporidesmium-like
K. dolioloides Switzerland Terrestrial Pinus Determined Undetermined
K. elaterascus Chile Freshwater Dead wood Determined Undetermined
K. emarceis Thailand Terrestrial Dead wood Determined Dendryphiopsis-like
K. fascicularis Unknown Unknown Unknown Undetermined Dendryphiopsis-like
K. fluminicola China Freshwater Submerged wood Undetermined Sporidesmium-like
K. goaensis India Terrestrial Bark of tree Undetermined Dendryphiopsis-like
K. incrustans Taiwan, China Terrestrial Dead wood Determined Dendryphiopsis-like
K. lignicola Thailand Terrestrial Dead wood Determined Dendryphiopsis-like
K. maritima U.S.A Marine On testblocks and also driftwood Determined Undetermined
......continued on the nextx page

KIRSCHSTEINIOTHELIA THAILANDICA SP. NOV. Phytotaxa 490 (2) © 2021 Magnolia Press • 179
TABLE 3. (Continued)
Taxon Country Habitat Host Sexual morph Asexual morph
K. phileura Unknown Unknown Unknown Determined Undetermined
K. phoenicis Thailand Terrestrial Phoenix paludosa Determined Undetermined
K. populi U.S.A Terrestrial Populus angustifolia Determined Undetermined
K. proteae South Africa Terrestrial Protea cynaroides Determined Undetermined
K. recessa Italy/U.S.A Terrestrial Pyrus/ dead wood Determined Dendryphiopsis-like
K. reticulata Taiwan, China Terrestrial Unknown twigs Determined Undetermined
K. rostrata Thailand Freshwater Submerged wood Undetermined Sporidesmium-like
K. smilacis Taiwan, China Terrestrial Smilax Determined Undetermined
K. striatispora Switzerland Terrestrial Juniperus communis Determined Undetermined
K. submersa China Freshwater Submerged wood Undetermined Sporidesmium-like
K. tectonae Thailand Terrestrial Tectona grandis Undetermined Sporidesmium-like
K. thailandica Thailand Terrestrial Ficus microcarpa Undetermined Sporidesmium-like
K. thujina Canada/U.S.A Terrestrial Abies balsamea/
Thuja occidentalis Determined Undetermined
K. umbrinoidea Italy Terrestrial Bark of aesculus hippocastanum Determined Undetermined
K. xera U.S.A Terrestrial Bark of prunus Determined Undetermined
Acknowledgements
We would like to thank the director of the Mae Fah Luang University botanical garden, the botanist Dr. Jantrararuk
Tovaranonte for her support given. We also want to thank Dr. Shaun Pennycook for checking the nomenclature. Kevin
D. Hyde would like to thank the Thailand Research grant entitled “Impact of climate change on fungal diversity and
biogeography in the Greater Mekong Subregion” (grant no: RDG6130001). The following projects supported the
research: National Natural Science Foundation of China (No. 31972222, 31560489), Program of Introducing Talents
of Discipline to Universities of China (111 Program, D20023), Talent project of Guizhou Science and Technology
Cooperation Platform ([2017]5788-5, [2019]5641 and [2020]5001) and Guizhou Science, Technology Department
International Cooperation Basic project ([2018]5806).
References
Bao, D.F., Luo, Z.L., Liu, J.K., Bhat, D.J., Sarunya, N., Li, W.L., Su, H.Y. & Hyde, K.D. (2018) Lignicolous freshwater fungi in China III:
Three new species and a new record of Kirschsteiniothelia from northwestern Yunnan Province. Mycosphere 9: 755–768.
https://doi.org/10.5943/mycosphere/9/4/4
Barr, M.E. (1987) Prodomus to class Loculoascomycetes. Amherst, Massachusetts, 168 pp.
Barr, M.E. (1993) Notes on the Pleomassariaceae. Mycotaxon 49: 129–142.
Boonmee, S., Ko, T.W.K., Chukeatirote, E., Hyde, K.D., Chen, H., Cai, L., McKenzie, E.H.C., Jones, E.B.G., Kodsueb, R. & Hassan, B.A.
(2012) Two new Kirschsteiniotheli a species with Dendryphiopsis anamorphs cluster in Kirschsteiniotheliaceae fam. nov. Mycologia
104: 698–714.
https://doi.org/10.3852/11-089
Capella-Gutiérrez, S., Silla-Martínez, J.M. & Gabaldón, T. (2009) trimAl: a tool for automated alignment trimming in large-scale
phylogenetic analyses. Bioinformatics 25: 1972–1973.
https://doi.org/10.1093/bioinformatics/btp348

SUN ET AL.
180 • Phytotaxa 490 (2) © 2021 Magnolia Press
Chernomor, O., Von Haeseler, A. & Minh, B.Q. (2016) Terrace aware data structure for phylogenomic inference from supermatrices.
Systematic Biology 65: 997–1008.
https://doi.org/10.1093/sysbio/syw037
Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature
Methods 9: 772–772.
https://doi.org/10.1038/nmeth.2109
Hawksworth, D.L. (1985) Kirschsteiniothelia, a new genus for the Microthelia incrustans-group (Dothideales). Botanical Journal of the
Linnean Society 91: 181–202.
https://doi.org/10.1111/j.1095-8339.1985.tb01144.x
Hernandez-Restrepo, M., Gené, J., Castañeda-Ruiz, R.F., Mena-Portales, J., Crous, P.W. & Guarro, J. (2017) Phylogeny of saprobic
microfungi from Southern Europe. Studies in Mycology 86: 53–97.
http://doi.org/10.1016/j.simyco.2017.05.002
Hillis, DM. & Bull, J.J. (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic
Biology 42: 182–192.
https://doi.org/10.1093/sysbio/42.2.182
Hongsanan, S., Hyde, K.D., Phookamsak, R., Wanasinghe, D.N., McKenzie, E.H.C., Sarma, V.V., Boonmee, S., Lücking, R., Bhat, D.J.,
Liu, N.G., Tennakoon, D.S., Pem, D., Karunarathna, A., Jiang, S.H., Jones, E.B.G., Phillips, A.J.L., Manawasinghe, I.S., Tibpromma,
S., Jayasiri, S.C., Sandamali, D.S., Jayawardena, R.S., Wijayawardene, N.N., Ekanayaka, A.H., Jeewon, R., Lu, Y.Z., Dissanayake,
A.J., Zeng, X.Y., Luo, Z.L., Tian, Q., Phukhamsakda, C., Thambugala, K.M., Dai, D.Q., Chethana, K.W.T., Samarakoon, M.C., Ertz,
D., Bao, D.F., Doilom, M., Liu, J.K., Pérez-Ortega, S., Suija, A., Senwanna, C., Wijesinghe, S.N., Konta, S., Niranjan, M., Zhang,
S.N., Ariyawansa, H.A., Jiang, H.B., Zhang, J.F., Norphanphoun, C., de Silva, N.I., Thiyagaraja, V., Zhang, H., Bezerra, J.D.P.,
Miranda-González, R., Aptroot, A., Kashiwadani, H., Harishchandra, D., Sérusiaux, E., Aluthmuhandiram, J.V.S., Abeywickrama,
P.D., Devadatha, B., Wu, H.X., Moon, K.H., Gueidan, C., Schumm, F., Bundhun, D., Mapook, A., Monkai, J., Chomnunti, P.,
Suetrong, S., Chaiwan, N., Dayarathne, M.C., Yang, J., Rathnayaka, A.R., Bhunjun, C.S., Xu, J.C., Zheng, J.S., Liu, G., Feng, Y. &
Xie, N. (2020) Refined families of Dothideomycetes: Dothideomycetidae and Pleosporomycetidae. Mycosphere 11: 1553–2107.
https://doi:10.5943/mycosphere/11/1/13
Hyde, K.D., Jones, E.B.G., Liu, J.K., Ariyawansa, H., Boehm, E., Boonmee, S., Braun, U., Chomnunti, P., Crous, P.W., Dai, D.Q.,
Diederich, P., Dissanayake, A., Doilom, M., Doveri, F., Hongsanan, S., Jayawardena, R., Lawrey, J.D., Li, Y.M., Liu, Y.X., Lücking,
R., Monkai, J., Muggia, L., Nelsen, M.P., Pang, K.L., Phookamsak, R., Senanayake, I.C., Shearer, C.A., Suetrong, S., Tanaka, K.,
Thambugala, K.M., Wijayawardene, N.N., Wikee, S., Wu, H.X., Zhang, Y., Aguirre-Hudson, B., Alias, S.A., Aptroot, A., Bahkali,
A.H., Bezerra, J.L., Bhat, D.J., Camporesi, E., Chukeatirote, E., Gueidan, C., Hawksworth, D.L., Hirayama, K., De Hoog, S., Kang,
J.C., Knudsen, K., Li, W.J., Li, X.H., Liu, Z.Y., Mapook, A., McKenzie, E.H.C., Miller, A.N., Mortimer, P.E., Phillips, A.J.L., Raja,
H.A., Scheuer, C., Schumm, F., Taylor, J.E., Tian, Q., Tibpromma, S., Wanasinghe, D.N., Wang, Y., Xu, J.C., Yacharoen, S., Yan, J.Y.
& Zhang, M. (2013) Families of Dothideomycetes. Fungal Diversity 63: 1–313.
https://doi:10.1007/s13225-013-0263-4
Hyde, K.D., Norphanphoun, C., Abreu, V.P., Bazzicalupo, A., Thilini, Chethana, K.W., Clericuzio, M., Dayarathne, M.C., Dissanayake,
A.J., Ekanayaka, A.H., He, M.Q., Hongsanan, S., Huang, S.K., Jayasiri, S.C., Jayawardena, R.S., Karunarathna, A., Konta, S.,
Kušan, I., Lee, H., Li, J.F., Lin, C.G., Liu, N.G., Lu, Y.Z., Luo, Z.L., Manawasinghe, I.S., Mapook, A., Perera, R.H., Phookamsak, R.,
Phukhamsakda, C., Siedlecki, I., Soares, A.M., Tennakoon, D.S., Tian, Q., Tibpromma, S., Wanasinghe, D.N., Xiao, Y.P., Yang, J.,
Zeng, X.Y., Abdel-Aziz, F.A., Li, W.J., Senanayake, I.C., Shang, Q.J., Daranagama, D.A., de Silva, N.I., Thambugala, K.M., Abdel-
Wahab ,M.A., Bahkali, A.H., Berbee, M.L., Boonmee, S., Bhat, D.J., Bulgakov, T..S, Buyck, B., Camporesi, E., Castañeda-Ruiz,
R.F., Chomnunti, P., Doilom, M., Dovana, F., Gibertoni, T.B., Jadan, M., Jeewon, R., Jones, E.B.G., Kang, J.C., Karunarathna, S.C.,
Lim, Y.W., Liu, J.K., Liu, Z.Y., Plautz, H..L, Lumyong, S., Maharachchikumbura, S.S.N., Matočec, N., McKenzie, E.H.C., Mešić,
A., Miller, D., Pawłowska, J., Pereira, O.L., Promputtha, I., Romero, A.I., Ryvarden, L., Su, H.Y., Suetrong, S., Tkalčec, Z., Vizzini,
A., Wen, T.C., Wisitrassameewong, K., Wrzosek, M., Xu, J.C., Zhao, Q., Zhao, R.L & Mortimer, P.E. (2017) Fungal diversity notes
603–708: taxonomic and phylogenetic notes on genera and species. Fungal Diversity 87: 1–235.
https://doi:10.1007/s13225-017-0391-3
Hyde, K.D., Norphanphoun, C., Chen, J., Dissanayake, A.J., Doilom, M., Hongsanan, S., Jayawardena, R.S., Jeewon, R., Perera, R.H.,
Thongbai, B., Wanasinghe, D.N., Wisitrassameewong, K., Tibpromma, S. & Stadler, M. (2018) Thailand’s amazing diversity: up to
96% of fungi in northern Thailand may be novel. Fungal diversity 93:215-239.
https://doi.org/10.1007/s13225-018-0415-7
Hyde, K.D., Jeewon, R., Chen, Y.J., Bhunjun, C.S., Calabon, M.S., Jiang, H.B., Lin, C.G., Norphanphoun, C., Sysouphanthong, P.,
Pem, D., Tibpromma, S., Zhang, Q., Doilom, M., Jayawardena, R.S., Liu, J.K., Maharachchikumbura, S.S.N., Phukhamsakda, C.,
Phookamsak, R., Al-Sadi, A.M., Thongklang, N., Wang, Y., Gafforov, Y., Jones, E.B.G. & Lumyong, S. (2020) The numbers of fungi:

KIRSCHSTEINIOTHELIA THAILANDICA SP. NOV. Phytotaxa 490 (2) © 2021 Magnolia Press • 181
is the descriptive curve flattening? Fungal Diversity 103: 219–271.
https://doi.org/10.1007/s13225-020-00458-2
Jayasiri, S.C., Hyde, K.D., Ariyawansa, H.A., Bhat, D.J., Buyck, B., Cai, L., Dai, Y.C., Abd-Elsalam, K.A., Ertz, D., Hidayat, I., Jeewon,
R., Jones, E.B.G., Bahkali, A.H., Karunarathna, S.C., Liu, J.K., Luangsa-ard, J.J., Lumbsch, H.T., Maharachchikumbura, S.S.N.,
McKenzie, E.H.C., Moncalvo, J.M., Ghobad-Nejhad, M., Nilsson, H., Pang, K.L., Pereira, O.L., Phillips, A.J.L., Raspé, O, Rollins,
A.W., Romero, A.I., Etayo, J., Selçuk, F., Stephenson, S.L., Suetrong, S., Taylor, J.E., Tsui, C.K.M., Vizzini, A., Abdel-Wahab,
M.A., Wen, T.C., Boonmee, S., Dai, D.Q., Daranagama, D.A., Dissanayake, A.J., Ekanayaka, A.H., Fryar, S.C., Hongsanan, S,
Jayawardena, R.S., Li, W.J., Perera, R.H., Phookamsak, R., de Silva, N.I., Thambugala, K.M., Tian, Q., Wijayawardene, N.N., Zhao,
R.L., Zhao, Q., Kang, J.C. & Promputtha, I. (2015) The faces of fungi database: fungal names linked with morphology, phylogeny
and human impacts. Fungal Diversity 74: 3–18.
https://doi.org/10.1007/s13225-015-0351-8
Jiang, S.H., Hawksworth, D.L., Lücking, R. & Wei, J.C. (2020) A new genus and species of foliicolous lichen in a new family of Strigulales
(Ascomycota: Dothideomycetes) reveals remarkable class-level homoplasy. IMA Fungus 11: 1
https://doi.org/10.1186/s43008-019-0026-2
Katoh, K. & Standley, D.M. (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability.
Molecular Biology and Evolution 30: 772–780.
https://doi.org/10.1093/molbev/mst010
Larsson, A. (2014) AliView: a fast and lightweight alignment viewer and editor for large datasets. Bioinformatics 30: 3276–3278.
https://doi.org/10.1093/bioinformatics/btu531
Lecocq, T., Vereecken, N.J., Michez, D., Dellicour, S., Lhomme, P., Valterova, I., Rasplus, J.Y. & Rasmont, P. (2013) Patterns of genetic
and reproductive traits differentiation in mainland vs. Corsican populations of bumblebees. PLoS One 8: e65642.
https://doi:10.1371/journal.pone.0065642
Li, G.J., Hyde, K.D., Zhao, R.L., Hongsanan, S., Abdel-Aziz, F.A., Abdel-Wahab, M.A., Alvarado, P., Alves-Silva, G., Ammirati, J.F.,
Ariyawansa, H.A., Baghela, A., Bahkali, A.H., Beug, M., Bhat, D.J., Bojantchev, D., Boonpratuang, T., Bulgakov, T.S., Camporesi,
E., Boro, M.C., Ceska, O., Chakraborty, D., Chen, J.J., Chethana, K.W.T., Chomnunti, P., Consiglio, G., Cui, B.K., Dai, D.Q., Dai,
Y.C., Daranagama, D.A., Das, K., Dayarathne, M.C., De Crop, E., De Oliveira, R.J.V., de Souza, C.A.F., de Souza, J.I., Dentinger,
B.T.M., Dissanayake, A.J., Doilom, M., Drechsler-Santos, E.R., Ghobad-Nejhad, M., Gilmore, S.P. & Góes-Neto, A. (2016) Fungal
diversity notes 253–366: taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 78: 1–237.
https://doi:10.1007/s13225-016-0366-9
Long, H., Zhang, Q., Hao, Y.Y., Shao, X.Q., Wei, X.X., Hyde, K.D., Wang, Y. & Zhao, D.G. (2019) Diaporthe species in south-western
China. MycoKeys 57: 113–127.
https://doi.org/10.3897/mycokeys.57.35448
Mehrabi, M., Hemmati, R. & Asgari, B. (2017) Kirschsteiniothelia arasbaranica sp. nov., and an emendation of the Kirschsteiniotheliaceae.
Cryptogamie, Mycologie 38: 13–25.
https://doi:10.7872/crym/v38.iss1.2017.13
Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In:
Proceeding of the 2010 gateway computing environments workshop (GCE), New Orleans, Louisiana, pp. 1–8.
https://doi.org/10.1109/ GCE.2010.5676129
Nguyen, L., Schmidt, H.A., Von Haeseler, A. & Minh, B.Q. (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating
maximum-likelihood phylogenies. Molecular Biology and Evolution 32: 268–274.
https://doi.org/10.1093/molbev/msu300
Nylander, J.A.A. (2004) MrModeltest 2.2. Department of Systematic Zoology, Uppsala University, Uppsala, Sweden.
Pratibha, J., Raghukumar, S. & Bhat, D.J. (2010) New species of Dendryphiopsis and Stauriella from Goa, India. Mycotaxon. 113:
297–303.
https://doi.org/10.5248/113.297
Rambaut, A. (2009) FigTree: Tree figure drawing tool, version 1. 4. 4. Institute of Evolutionary Biology, University of Edinburgh.
Rannala, B. & Yang, Z.H (1996) Probability distribution of molecular evolutionary trees: A new method of phylogenetic inference.
Journal of Molecular Evolution 43: 304–311.
https://doi.org/10.1007/BF02338839.
Ronquist, F., Teslenko, M., Van Der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P.
(2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology
61: 539–542.
https://doi.org/10.1093/sysbio/sys029

SUN ET AL.
182 • Phytotaxa 490 (2) © 2021 Magnolia Press
Schoch, C.L., Shoemaker, R.A., Seifert, K.A., Hambleton, S., Spatafora, J.W. & Crous, P.W. (2006) A multigene phylogeny of the
Dothideomycetes using four nuclear loci. Mycologia 98: 1041–1052.
https://doi.org/10.1080/15572536.2006.11832632
Schoch, C., Crous, P.W., Groenewald, J.Z., Boehm, E., Burgess, T.I., De Gruyter, J., De Hoog, G.S., Dixon, L., Grube, M. & Gueidan, C.
(2009) A class-wide phylogenetic assessment of Dothideomycetes. Studies in Mycology 64: 1–15.
https://doi.org/10.3114/sim.2009.64.01
Swofford, D.L. (2002) PAUP*: phylogenetic analysis using parsimony (and other methods) version 4.0 b10. MA: Sinauer Associates,
Sunderland, UK.
Su, H.Y., Hyde, K.D., Maharachchikumbura, S.S.N., Ariyawansa, H.A., Luo, Z.L., Promputtha, I., Tian, Q., Lin, C.G., Shang, Q.J., Zhao,
Y.C., Chai, H.M., Liu, X.Y., Bahkali, A.H., Bhat, J.D., McKenzie, E.H.C. & Zhou, D.Q. (2016) The families Distoseptisporaceae
fam. nov., Kirschsteiniotheliaceae, Sporormiaceae and Torulaceae, with new species from freshwater in Yunnan Province, China.
Fungal Diversity 80: 375–409.
https://doi:10.1007/s13225-016-0362-0
Suetrong, S., Schoch, C.L., Spatafora, J.W., Kohlmeyer, J., Volkmann-Kohlmeyer, B., Sakayaroj, J., Phongpaichit, S., Tanaka, K.,
Hirayama, K. & Jones, E.B.G. (2009) Molecular systematics of the marine Dothideomycetes. Studies in Mycology 64: 155-173.
https://doi.org/10.3114/sim.2009.64.09
Sun, Y.R., Goonasekara, I.D., Thambugala, K.M., Jayawardena, R.S., Wang, Y. & Hyde, K.D. (2020) Distoseptispora bambusae sp.
nov.(Distoseptisporaceae) on bamboo from China and Thailand. Biodiversity Data Journal 8: e53678.
https://doi:10.3897/BDJ.8.e53678
Vaidya, G., Lohman, D.J. & Meier, R. (2011) SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with
character set and codon information. Cladistics 27: 171–180.
https://doi.org/10.1111/j.1096-0031.2010.00329.x
Vilgalys, R. & Hester, M. (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several
Cryptococcus species. Journal of Bacteriology 172: 4238–4246.
White, T.J., Bruns, T., Lee, S. & Taylor, J. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.
In: Innis, M., Gelfand, D., Shinsky, J. & White, T. (Eds.) PCR protocols: a guide to methods and applications. Academic Press, New
York, pp. 315−322.
https://doi.org/10.1016/B978-0-12-372180-8.50042-1
Wijayawardene, N.N., Crous, P.W., Kirk, P.M., Hawksworth, D.L., Boonmee, S., Braun, U., Dai, D.Q., D’souza, M.J., Diederich, P.,
Dissanayake, A., Doilom, M., Hongsanan, S., Jones, E.B.G., Groenewald, J.Z., Jayawardena, R., Lawrey, J.D., Liu, J.K., Lücking,
R., Madrid, H., Manamgoda, D.S., Muggia, L., Nelsen, M.P., Phookamsak, R., Suetrong, S., Tanaka, K., Thambugala, K.M.,
Wanasinghe, D.N., Wikee, S., Zhang, Y., Aptroot, A., Ariyawansa, H.A., Bahkali, A.H., Bhat, D.J., Gueidan, C., Chomnunti, P., De
Hoog, G.S., Knudsen, K., Li, W.J., McKenzie, E.H.C., Miller, A.N., Phillips, A.J.L., Piątek, M., Raja, H.A., Shivas, R.S., Slippers,
B., Taylor, J.E., Tian, Q., Wang, Y., Woudenberg, J.H.C., Cai, L., Jaklitsch, W.M. & Hyde, K.D. (2014) Naming and outline of
Dothideomycetes–2014 including proposals for the protection or suppression of generic names. Fungal Diversity 69: 1–55.
https://doi:10.1007/s13225-014-0309-2
Wijesinghe, S.N., Wang, Y., Camporesi, E., Wanasinghe, D.N., Boonmee, S. & Hyde, K.D. (2020) A new genus of Bambusicolaceae
(Pleosporales) on Corylus avellana (Fagales) from Italy. Biodiversity Data Journal 8: e55957.
https://doi:10.3897/BDJ.8.e55957
Zhang, Q., Yang, Z.F., Cheng, W., Wijayawardene, N.N., Hyde, K.D., Chen, Z. & Wang, Y. (2020) Diseases of Cymbopogon citratus
(Poaceae) in China: Curvularia nanningensis sp. nov. MycoKeys 63: 49–67.
https://doi: 10.3897/mycokeys.63.49264