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Phytotaxa 516 (1): 001–027
https://www.mapress.com/j/pt/
Copyright © 2021 Magnolia Press Article PHYTOTAXA
ISSN 1179-3155 (print edition)
ISSN 1179-3163 (online edition)
Accepted by Jian-Kui Liu: 22 Jun. 2021; published: 20 Aug. 2021
https://doi.org/10.11646/phytotaxa.516.1.1
1
First reports of the sexual morphs of Diaporthe forlicesenica nom. nov. and
Diaporthe goulteri (Diaporthaceae, Diaporthales) revealed by molecular
phylogenetics
DIGVIJAYINI BUNDHUN1,2,10, INDUNIL C. SENANAYAKE3,4,11, RUVISHIKA S. JAYAWARDENA2,12, ERIO
CAMPORESI5,6,7,13, YINGHUA HUANG4,14, ZHANGYONG DONG4,15, KEVIN D. HYDE1,2,4,16, CHAIWAT TO-
ANUN1,17 & RATCHADAWAN CHEEWANGKOON1,8,9,18*
1 Department of Entomology and Plant Pathology, Faculty of Agriculture, Chiang Mai University, Chiang Mai, 50200, Thailand
2 Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand
3 College of Life Science and Oceanography, Shenzhen University, 1068, Nanhai Avenue, Nanshan, Shenzhen 518055, People’s Republic
of China
4 Innovative Institute for Plant Health, Zhongkai University of Agriculture and Engineering, Haizhu District, Guangzhou, People’s
Republic of China
5 A.M.B. Gruppo Micologico Forlivese “Antonio Cicognani”, Via Roma 18, Forli, Italy
6 A.M.B, Circolo Micologico “Giovanni carini”, C.P. 314, Brescia, Italy
7 Societa per gli Studi Naturalistici della Romagna, C.P. 144, Bagnacavallo, RA, Italy
8 Innovative Agriculture Research Center, Faculty of Agriculture, Chiang Mai University.
9 Center of Excellence in Microbial Diversity and Sustainable Utilization, Chiang Mai University, Chiang Mai, 50200, Thailand
10
digvi221993@gmail.com; https://orcid.org/0000-0002-0790-215X
11
indunilchinthani@gmail.com; https://orcid.org/0000-0002-7165-2394
12
ruvi.jaya@yahoo.com; https://orcid.org/0000-0001-7702-4885
13
eriocamporesi@libero.it; https://orcid.org/0000-0002-2801-9462
14
1065488893@qq.com; https://orcid.org/0000-0003-4599-6430
15
dongzhangyong@hotmail.com; https://orcid.org/0000-0001-7524-0226
16
kdhyde3@gmail.com; https://orcid.org/0000-0002-2191-0762
17
chaiwat.toanun@gmail.com; https://orcid.org/0000-0003-3772-235X
18
ratchadawan.c@cmu.ac.th; https://orcid.org/0000-0001-8576-3696
* Corresponding author: ratchadawan.c@cmu.ac.th
Abstract
Diaporthe forlicesenica nom. nov. is proposed for D. dorycnii Dissan., Camporesi & K.D. Hyde, a later homonym of D.
dorycnii (Mont.) Sacc. Diaporthe forlicesenica as well as the species D. goulteri have so far only been described in their
asexual morphs. In this study, the sexual morphs for these species are recovered for the first time, from the dead branches
of Cytisus sp. in Italy and from an unknown host in Thailand. The asexual-sexual morph connections of the species are con-
firmed by DNA sequence based phylogenetic analyses including the ITS, tef1, tub2 and his loci, supported by morphology.
Detailed descriptions, illustrations and molecular data for the taxa are provided.
Keywords: Diaporthomycetidae, Fungal diversity, Phylogeny, Saprobe, Sordariomycetes, Taxonomy
Introduction
Diaporthe Nitschke. is typified by D. eres Nitschke (1870). Species of Diaporthe occur as endophytes, pathogens or
saprobes worldwide, on a wide variety of plants and even on submerged wood in freshwater habitats (Thompson et al.
2011, 2015, Dissanayake et al. 2017a,b, 2020, Calabon et al. 2020). Asexual morphs of Diaporthe taxa were earlier
referred to as Phomopsis (Rehner & Uecker 1994, Santos et al. 2010, Udayanga et al. 2011, 2012). Species could,
therefore, potentially have dual nomenclature. Both morphs of the same species are seldom discovered simultaneously.
This implies that two different names were being given to the same species, one for its sexual morph and another for its
asexual morph. However, in accordance with the International Code of Nomenclature, which required the eradication
of dual nomenclature for pleomorphic taxa, the older name Diaporthe was chosen over Phomopsis (McNeill et al.
BUNDHUN ET AL.
2 • Phytotaxa 516 (1) © 2021 Magnolia Press
2012, Rossman et al. 2015), which has been followed in the latest outline of the fungi (Wijayawardene et al. 2020).
Linkage of sexual-asexual morph essentially reduces an unnecessary amount of fungal names while equally promoting
a more comprehensive fungal classification (Wingfield et al. 2012). Likewise, several Diaporthe species have been
linked to their Phomopsis taxa (Udayanga et al. 2012, 2014, Rossman et al. 2015). Incorporation of DNA sequence data
has currently become instrumental in linking sexual and asexual morphs of Diaporthe taxa (Yang et al. 2017, Perera
et al. 2018). Molecular based phylogeny becomes particularly important in cases where morphological comparison
for same species cannot be easily made. The morphology may vary based on the culture media, age of sporulation,
intra-species variability or if the morphs being compared have been retrieved from different sources (that is, culture or
natural host) (Hyde et al. 2019, Senanayake et al. 2020).
Diaporthe is a speciose genus and over the years, as taxa have been introduced, some have inadvertently been
invalidly established; for instance, D. interrupta Niessl and D. dorycnii Dissan., Camporesi & K.D. Hyde (Index
Fungorum 2021). Diaporthe interrupta Niessl (1883) (Berlese & Voglino 1886) was introduced with similar epithet
as D. interrupta (Mont.) Sacc (1882) (Saccardo 1882a). Diaporthe tecomae Sacc. & P. Syd. was then proposed for D.
interrupta Niessl (Saccardo 1899, Gomes et al. 2013) since two species within the same genus cannot have identical
epithets. Indeed, according to the Article 53 (53.1) in International Code of Nomenclature for algae, fungi, and plants
(Melbourne Code), “a name of a family, genus, or species, unless conserved (Art. 14) or sanctioned (Art. 15), is
illegitimate if it is a later homonym, that is, if it is spelled exactly like a name based on a different type that was
previously and validly published for a taxon of the same rank” (McNeill et al. 2012). Likewise, D. dorycnii Dissan.,
Camporesi & K.D. Hyde (2017) is a later homonym of D. dorycnii (Mont.) Sacc. (1882) (Saccardo 1882b, Dissanayake
et al. 2017b).
In an ongoing study of Sordariomycetes, two taxa of Diaporthe in their sexual morphs have been collected,
isolated and analyzed. One strain is identified as ‘D. dorycnii’ Dissan., Camporesi & K.D. Hyde while the other one
as D. goulteri R.G. Shivas, S.M. Thomps. & Y.P. Tan. These two species have been described solely in their asexual
morphs until now (Thompson et al. 2015, Dissanayake et al. 2017b,c). The aims of the current study are to firstly,
resolve the nomenclatural error vis-à-vis ‘D. dorycnii’ by proposing D. forlicesenica Bundhun, Camporesi & K.D.
Hyde nom. nov. and secondly, to report the sexual morphs of D. forlicesenica and D. goulteri for the first time, based
on multi-locus sequence analyses and morphological characterization.
Materials and Methods
Sample collection, specimen examination and isolation
Dead branches were collected from Cytisus sp. in Forlì-Cesena, Italy in 2018 and from an unknown host in Chiang Rai,
Thailand in 2019. The samples were taken to the laboratory in a plastic Ziplock bag and stored inside paper envelopes.
Specimens were externally examined with a Motic SMZ 168 stereomicroscope. Free-hand sections of ascomata were
prepared and examined using a Nikon ECLIPSE 80i compound microscope. Microscopic photography was carried out
and images were captured with a Canon EOS 750D digital camera. Measurements were made with the Tarosoft (R)
Image Frame Work version 0.9.7. and images used for figures were processed with Adobe Photoshop CS6 Extended
version 13.0.1 software (Adobe Systems, San Jose, California).
Single spore isolation was carried out following the method of Senanayake et al. (2020). Germinated ascospores
on potato dextrose agar (PDA) were examined after 24 h and they were transferred to PDA media. Cultures were
incubated at 25 °C in the dark and colony color was examined according to Rayner (1970) after 1, 3 and 4 weeks of
growth on PDA. Herbarium specimens are deposited in Mae Fah Luang University Herbarium (MFLU) while, living
cultures, at the Mae Fah Luang University Culture Collection (MFLUCC) in Thailand. Index Fungorum and Faces of
Fungi numbers are provided as per Index Fungorum (2021) and Jayasiri et al. (2015).
DNA extraction, PCR amplification and sequencing
Total DNA was extracted from aerial mycelium of cultures grown on PDA at 25 °C for one week. The rDNA internal
transcribed spacer (ITS) region was initially amplified and sequenced to identify the fungus to genus level. Following
this, translation elongation factor-1α (tef1), partial β-tubulin (tub2), partial histone H3 (his) and calmodulin (cal) genes
were sequenced to support species identification initially revealed through the ITS sequence data. Primer pairs and
annealing temperature used for each gene region are given in TABLE 1. However, sequence data for cal could not be
obtained despite several trials using different temperatures (temperature gradient).
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 3
TABLE 1. Primer pairs and annealing temperatures for gene regions sequenced in the present study.
Gene region Primer pair Annealing temperature Reference
ITS ITS5/ ITS4 55 °C / 50s White et al. 1990
tef1 EF-728F/ EF-986R 58 °C / 30s Carbone & Kohn 1999
tub2 BT2a/ BT2b 58 °C / 50s Glass & Donaldson 1995, Udayanga et al. 2012
his CYLH3F/ H3-1b 58 °C / 50s Glass & Donaldson 1995, Crous et al. 2004
cal CAL228F/ CAL737R 55 °C / 50s Carbone & Kohn 1999
PCR mixtures of 25 μl total volume consisted of 0.3 μl of TaKaRa Ex-Taq DNA polymerase, 2.5 μl of 10 × Ex-
Taq DNA polymerase buffer, 3.0 μl of dNTPs, 2 μl of genomic DNA, 1 μl of each primer, and 15.2 μl ddH2O. PCR
products were verified by staining with ethidium bromide on 1% agarose electrophoresis gels. They were then purified
and sequenced using the same primers at Tianyi Huiyuan Biotechnology Co., Ltd., Guangzhou, China. The sequence
quality was confirmed by checking chromatograms using BioEdit v. 7.0 (Hall 2004) and consensus sequence for each
gene region was assembled in SeqMan (DNASTAR software version 5.0; DNASTAR Inc., Madison, WI). Nucleotide
sequences derived in this study have been deposited in GenBank (Supplementary TABLE 1).
Phylogenetic analyses
Generated ITS, tef1, tub2 and his sequences were subjected to BLASTn searches (https://blast.ncbi.nlm.nih.gov) and
related sequences were downloaded from GenBank (Supplementary TABLE 1). Individual gene matrices were aligned
using default setting in MAFFT V.7.036 (http://mafft.cbrc.jp/alignment/server/) (Katoh et al. 2019) and manually
improved when necessary in BioEdit v. 7.0 (Hall 2004). Absent sequences were coded as missing data and characters
were assessed to be unordered and equally weighted.
Maximum likelihood (ML) and Bayesian posterior probability (BYPP) analyses were conducted using both
individual and combined datasets of ITS, tef1, tub2, his as well as cal. An initial phylogenetic tree, comprising
representative strains of all Diaporthe taxa with molecular data, including our strains, was constructed based on a
combined dataset of the five loci (Supplementary FIGURE 1). Considering the placement of our strains in the initial
tree, a second tree was re-constructed using taxa from the subclades where our strains grouped along with representative
strains from other subclades. Diaporthella corylina (CBS 121124) was used as the outgroup taxon.
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/) bioinformatics web tool (Glez-Pea et al.
2010). They were then used to generate ML trees using RAxML-HPC2 on XSEDE (v.8.2.10) (Stamatakis 2014) with
the GTRGAMMA substitution model and bootstrapping with 1,000 replicates.
The BYPP analysis was generated using Markov Chain Monte Carlo sampling in MrBayes v3.1.2 (Huelsenbeck
& Ronquist 2001, Zhaxybayeva & Gogarten 2002). MrModeltest v.2.3 (Nylander 2004) was used to estimate the
best evolutionary models for each gene region under the Akaike Information Criterion (AIC) implemented in PAUP
v.4.0b10 (Swofford 2003). Six simultaneous Markov chains were run for 10M (Supplementary FIGURE 1) and 4M
(FIGURE 1) generations with trees sampled every 100th generation. The distribution of log-likelihood scores was
examined using Tracer 1.5 (Rambaut & Drummond 2007) in order to determine the stationary phase for each search
and decide whether additional runs were required to achieve convergence. The first 20% of generated trees were the
burn-in phase and discarded. The remaining 80% of trees were used to calculate posterior probabilities in the majority
rule consensus tree. The resulting trees were viewed in FigTree v.1.4.0 (Rambaut 2012) and modified in Microsoft
PowerPoint (2013). The final alignments and phylogenetic tree were deposited in TreeBASE, submission ID 28005
(http://www.treebase.org).
Results
Phylogenetic analyses
The initial dataset for the overview phylogenetic tree, based on ITS, tef1, tub2, his and cal sequence data, comprised
273 taxa, including the two strains used in the present study (Diaporthe forlicesenica MFLUCC 21-0011 and D.
goulteri MFLUCC 21-0012). Diaporthella corylina (CBS 121124) was used as the outgroup taxon. A best scoring
RAxML tree with a final ML optimization likelihood value of -91990.044067 was yielded (Supplementary FIGURE
1).
BUNDHUN ET AL.
4 • Phytotaxa 516 (1) © 2021 Magnolia Press
FIGURE 1. RAxML tree based on analysis of a combined dataset of ITS, tef1, tub2, his and cal sequence data. Bootstrap support values
for ML values ≥ 50% and BYPP values ≥ 0.90 are shown as ML/BYPP above or below the branches respectively. The isolates generated in
the present study are shown in bold blue. Ex-types are indicated in bold black. The tree is rooted with Diaporthella corylina (CBS 121124).
The scale bar represents the expected number of nucleotide substitutions per site.
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 5
The re-constructed phylogenetic tree (FIGURE 1) based on a combined dataset of ITS, tef1, tub2, his and cal
sequence data, consisted of 94 taxa, including our strains and the outgroup Diaporthella corylina (CBS 121124). The
concatenated alignment consisted of 2,875 characters, including gaps (ITS: 546; tef1: 428; tub2: 843; his: 525; cal:
533). The ML tree topology was similar to the one of the BYPP consensuses trees. The best scoring RAxML tree with
final optimization had a likelihood value of -40381.734311. The matrix had 1,857 distinct alignment patterns, with
33.13% of gaps and completely undetermined characters. Estimated base frequencies were as follows: A= 0.211168,
C= 0.331128, G= 0.236331, T= 0.221373, with substitution rates AC= 1.003109, AG= 3.260196, AT= 1.087390, CG=
0.803975, CT= 4.305892, GT= 1.000000. The gamma distribution shape parameter α= 0.401292 and Tree-length=
5.919789.
The strain MFLUCC 21-0011 grouped with “D. dorycnii” (MFLUCC 17-1015) with high support values (100%
ML and 1.00 BYPP) while strain MFLUCC 21-0012 clustered with D. goulteri (BRIP 55657a) with 100% ML and
1.00 BYPP statistical support (FIGURE 1).
Taxonomy
Diaporthe forlicesenica Bundhun, Camporesi & K.D. Hyde, nom. nov. FIGURES 2 & 3
Basionym: Diaporthe dorycnii Dissan., Camporesi & K.D. Hyde, in Dissanayake, Camporesi, Hyde, Zhang, Yan & Li, Mycosphere 8(5):
867 (2017)
Index Fungorum number: IF558167, Facesoffungi number: FoF 03272
Etymology:— The specific epithet forlicesenica refers to Forlì-Cesena Province, where the fungus was collected.
Saprobic on dead aerial branches. Sexual morph: Ascomata 610–650 μm high, 215–370 μm diam. (x̅ = 630 × 310 μm,
n = 15), perithecial, black, globose to subglobose, scattered, gregarious, embedded in bark epidermis, with short black
cylindrical necks protruding through host surface, ostiolate. Neck 280–370 μm high, 70–170 μm diam. (x̅ = 305 × 133
μm, n = 15). Ostiole comprising hyaline periphyses. Peridium 2-layered; outer layer 10–20 μm wide, dense, composed
of dark brown thick-walled cells of textura angularis, inner layer 5–10 μm wide, comprising thin-walled, light brown
to hyaline cells of textura angularis. Paraphyses ca. 5 μm wide, hyaline, septate, constricted at the septa, guttulate,
thin-walled. Asci 50–70 × 5–15 μm (x̅ = 57.5 × 9.7 μm, n = 30), unitunicate, 8-spored, sessile, cylindrical to elongate,
apically truncate to rounded, with a non-amyloid refractive apical ring, straight or slightly curved. Ascospores 13–19
× 2–4 μm (x̅ = 15.7 × 3 μm, n = 30), uni- to bi-seriate, at times tri-seriate, hyaline, 1-septate, regularly tetra-guttulate,
with the two larger guttules in the middle and smaller ones at the ends, oblong or cylindrical to broadly fusiform,
slightly constricted at the septum, straight to slightly curved, with rounded to obtuse ends, smooth-walled, without
appendage or sheath. Asexual morph: Coelomycetous. Conidiomata up to 900 μm high and 1.0 mm in diam. on
PDA, superficial, pycnidial to multilocular, solitary or aggregated, scattered, globose to irregular, dark brown to black.
Conidiophores 10–30 × 1–3 μm (x̅ = 19 × 1.8 μm, n = 20), cylindrical, aseptate, rarely 1-septate, straight or sinuous,
terminal, slightly tapered towards the apex. Conidiogenous cells 3–15 × 1–2 μm (x̅ = 6 × 1.6 μm, n = 20) hyaline,
subcylindrical, filiform, tapering towards the apex. Alpha conidia 5–9 × 1–3 μm (x̅ = 7.4 × 2.4 μm, n = 30) hyaline,
often guttulate at maturity, fusiform or oval, both ends obtuse. Beta conidia not observed.
Culture characteristics:—Colonies on PDA, reaching 40 mm diam. after 1 week at 25 °C, initially white to light
brown, becoming dark brown with age on surface, reverse dark brown, smooth surface, entire margin. Sporulation
occurred on PDA after almost 45 days incubation period at 25 °C, in dark.
Material examined:—ITALY, Forlì-Cesena, Castagnolo, Civitella di Romagna, on dead aerial branches of Cytisus
sp. (Fabaceae), 17 December 2018, E. Camporesi, IT 4156 (MFLU 19-0348), living culture MFLUCC 21-0011.
Notes:—Diaporthe dorycnii was, upon its introduction, mistakenly given the name of an already existing epithet
in Diaporthe (Dissanayake et al. 2017b). It was therefore, invalidly published. One of our isolate MFLUCC 21-0011
is clustered with “D. dorycnii” MFLUCC 17-1015 with high statistical support (100% ML, 1.00 BYPP) (FIGURE 1).
Base pair (bp) comparison between ITS sequence data revealed 1 out of 472 (0.2%) difference in nucleotides while 11
out of 302 (3.6%) and 6 out of 376 (1.6%) bp differences in tef1 and tub2. Base pair differences in the latter two gene
regions arose mainly from the dissimilar bases at the start of each sequence. We refrain from counting them as reliable
differences since they are most probably the result of sequencing errors. According to the guidelines of Jeewon & Hyde
(2016), the genetic variations mentioned above are therefore insignificant to delineate the two isolates as different
species. Therefore, they are considered as one and we propose the new name D. forlicesenica to replace D. dorycnii
Dissan., Camporesi & K.D. Hyde.
BUNDHUN ET AL.
6 • Phytotaxa 516 (1) © 2021 Magnolia Press
FIGURE 2. Sexual morph of Diaporthe forlicesenica (MFLU 19-0348). a Specimen. b Ascomata on host substrate. c Close-up of
ascomata on host substrate. d Vertical section through ascomata (in KOH) e Section through the neck (in KOH). f Vertical section of
peridium. g Paraphyses. h–j Asci (j in Congo red). k Apical apparatus showing non-amyloid reaction in Melzer’s reagent. l–o Hyaline
ascospores, slightly constricted at the septum. Scale bars: b,c = 100 μm, d = 200 μm, e = 50 μm, f, l–o = 10 μm, g, k = 5 μm, h–j = 20
μm.
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 7
FIGURE 3. Asexual morph of Diaporthe forlicesenica (MFLUCC 21-0011). a Upper view of 45 days old colony on PDA. b Reverse
view of 45 days old colony on PDA. c Conidiomata in culture. d,e Conidiophores with alpha conidia. f–i Alpha conidia. Scale bars: d =
10 μm, e,g,i = 5 μm, f,h = 3 μm.
The strain MFLUCC 17-1015 of D. forlicesenica was described in its asexual morph from host substrate
(Dissanayake et al. 2017b) while our strain MFLUCC 21-0011 has presently been retrieved in both its sexual (natural
host) and asexual (culture on PDA) morphs. The conidiophores, conidiogenous cells and alpha conidia obtained from
the culture of MFLUCC 21-0011, however, are different in size from those of D. forlicesenica MFLUCC 17-1015
(TABLE 2).
TABLE 2. Morphological comparison between Diaporthe forlicesenica MFLUCC 21-0011 and MFLUCC 17-1015.
Features Diaporthe forlicesenica MFLUCC
21-0011 (this study) (μm)
Diaporthe forlicesenica MFLUCC 17-
1015 (Dissanayake et al. 2017b) (μm)
Conidiophores 10–30 × 1–3 21–35 × 1.5–2.5
Conidiogenous cells 3–15 × 1–2 13–19 × 2–3
Alpha conidia 5–9 × 1–3 9–13.5 × 3–4
BUNDHUN ET AL.
8 • Phytotaxa 516 (1) © 2021 Magnolia Press
Diaporthe goulteri R.G. Shivas, S.M. Thomps. & Y.P. Tan, Persoonia 35: 43 (2015) amend. FIGURES 4 & 5
Index Fungorum number: IF808669, Facesoffungi number: FoF 09714
Saprobic on wood litter. Sexual morph: Ascomata 360–480 μm high, 145–280 μm diam. (x̅ = 420 × 210 μm, n =
15), perithecial, black, globose to conical, scattered to aggregated, deeply immersed in host epidermis, with elongated
black necks emerging through host tissue, ostiolate. Neck 170–230 μm high, 45–70 μm diam. (x̅ = 190 × 55 μm, n =
15). Ostiole papillate, periphysate. Peridium 2-layered; outer layer 10–15 μm wide, heavily pigmented, composed of
dark brown, thick-walled cells of textura angularis, inner layer 10–20 μm wide, comprising light brown to hyaline,
thin-walled cells of textura angularis. Paraphyses 4–5 μm wide, hyaline, septate, wider at the base, thin-walled. Asci
30–50 × 5–10 μm (x̅ = 39.1 × 7.4 μm, n = 30), unitunicate, 8-spored, sessile, broadly cylindrical to sub-obclavate,
thin-walled, apically rounded, with a J-, apical ring, straight to slightly curved. Ascospores 7–10 × 2–4 μm (x̅ = 8.8
× 2.4 μm, n = 30), overlapping uniseriate to bi- or tri-seriate, hyaline, 1-septate, regularly 4-guttulate, with the two
larger guttules at the center and smaller ones at the ends, fusiform to elliptical, straight, smooth-walled, lacking any
appendage or mucilaginous sheath. Asexual morph: Coelomycetous. Conidiomata up to 600 μm high and 900 μm in
diam., erumpent to superficial on PDA, pycnidial to multilocular, scattered to aggregated, globose to irregular, black.
Conidiophores 15–20 × 2–3 μm (x̅ = 18.4 × 2.1 μm, n = 20), originating from the innermost layer of the locular wall,
1-septate or reduced to conidiogenous cells, hyaline, filiform, smooth-walled. Conidiogenous cells 3–8 × 1–2 μm (x̅
= 5 × 1.5 μm, n = 20), cylindrical, straight to flexuous, tapering towards the apex, hyaline. Alpha conidia 5–8 × 2–3
μm (x̅ = 6.9 × 2.1 μm, n = 30), produced in abundance, fusiform to cylindrical, rounded to slightly obtuse at the ends,
hyaline, smooth-walled. Beta conidia 15–20 × 1–2 μm (x̅ = 17 × 1.4 μm, n = 20) few, filiform, hooked, aseptate,
hyaline, smooth-walled. Gamma conidia not observed.
Culture characteristics:—Colonies on PDA, reaching 35 mm diam. after 1 week at 25 °C, flat, irregular, with a
smooth surface, entire margin, off-white to buff, reverse buff. Sporulation occurred on PDA after 30 days incubation
period at 25 °C, in dark.
Material examined:—THAILAND, Chiang Rai Province, Mae Fah Luang, on a dead branch of an unknown host,
11 May 2019, D. Bundhun, DB150 (MFLU 21-0022), living culture MFLUCC 21-0012.
Notes:—Diaporthe goulteri was introduced and described by Thompson et al. (2015) in its asexual morph. In
the present study, an isolate of Diaporthe (MFLUCC 21-0012) clustered with the strain of D. goulteri (BRIP 55657a)
with strong statistical support (100% ML, 1.00 BYPP) (FIGURE 1). The insignificant differences in base pairs in the
ITS, tef1 and tub2 sequence data for the two strains (<1.5%) support them as a single species (Jeewon & Hyde 2016).
Moreover, the asexual morph produced in culture (FIGURE 5) also corroborates with the morphology in Thompson
et al. (2015). The minor differences in size of the morphological characters may be accounted for by environmental
variations. To supplement the observation of Thompson et al. (2015), beta conidia of D. goulteri have also been
observed in culture in the present study. However, the attachment of these conidia on the conidiophores could not be
easily observed. Thus we were unable to get a clear photograph of the attachment.
Discussion
The introduction of molecular technique has enabled better resolution of taxa in Diaporthe, which has in turn allowed
establishment of the asexual-sexual morph connections for D. forlicesenica and D. goulteri in the present study.
Although the asexual features observed from the culture of D. forlicesenica MFLUCC 21-0011 were different in size
from those reported in Dissanayake et al. (2017b) (TABLE 2), this difference may have arisen from the fact that D.
forlicesenica MFLUCC 17-1015 was isolated from natural host (Dissanayake et al. 2017b) while the features were
obtained on PDA medium in this study. The addition of the sexual morphs of D. forlicesenica and D. goulteri expands
the morphology of the two taxa which were so far only known in their asexual morphs.
Diaporthe forlicesenica
was initially described from Italy, as saprobe on Dorycnium hirsutum (Dissanayake et al.
2017b). We recovered the species from the same country (and same province), with the same nutritional mode, but in
its sexual form and from another host, namely, Cytisus sp. Diaporthe goulteri was reported from Australia, from the
seed of Helianthus annuus as well as weed debris (Thompson et al. 2015). It was mentioned that this species could
colonize and survive on dead plant materials as saprobe between cropping phases until it colonized the next plantation
and potentially changed to pathogenic lifestyle. Sexual morphs of several plant pathogenic fungi have been reported
to have a high survival potential in challenging conditions (McDonald & Linde 2002). For instance, they can resist
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 9
FIGURE 4. Sexual morph of Diaporthe goulteri (MFLU 21-0022). a Specimen. b Ascomata on host substrate. c Section through
ascomata. d Section through the neck. e Vertical section of peridium. f Paraphyses. g–j Asci (i in Cotton blue reagent; j in Congo red). k
Apical apparatus showing non-amyloid reaction in Melzer’s reagent. l–o Ascospores. p Germinating ascospore. Scale bars: b,d = 50 μm,
c = 100 μm, e = 30 μm, f = 4 μm, g–j = 10 μm, k–p = 5 μm.
BUNDHUN ET AL.
10 • Phytotaxa 516 (1) © 2021 Magnolia Press
overwintering phases and later infect plants creating a new infection cycle (Schubert et al. 2003). This might also be
the case for D. goulteri during the cropping phase, whereby it may be present in its sexual form as well. However, no
such observation has been made or confirmed. There has been no report so far about the pathogenicity of D. goulteri
on any crop of importance in Thailand. However, this possibility cannot be discounted as its ecological significance in
this region still remains to be discovered.
FIGURE 5. Asexual morph of Diaporthe goulteri (MFLUCC 21-0012). a Upper view of 30 days old colony on PDA. b Reverse view
of 30 days old colony on PDA. c Conidiomata in culture. d,e Conidiophores with alpha conidia. f Conidiophore with beta conidia. g Alpha
conidia. h Beta conidium. Scale bars: d–f = 10 μm, g,h = 5 μm.
Acknowledgements
We thank the Thailand Research Fund, grant RDG6130001 entitled “Impact of climate change on fungal diversity
and biogeography in the Greater Mekong Subregion”. Kevin D. Hyde thanks Chiang Mai University for the award of
Visiting Professor. We would also like to thank S.C. Karunarathna’s grant, CAS President’s International Fellowship
Initiative (PIFI) under the following grant: 2018PC0006 and the National Science Foundation of China (NSFC,
project code 31750110478)”. Digvijayini Bundhun is grateful to Paul Kirk and Shaun Pennycook for nomenclature
verification. Milan C. Samarakoon, Pranami Abeywikrama and Chitrabhanu S. Bhunjun are thanked for their helpful
suggestions. Mae Fah Luang University, the Mushroom Research Foundation and the Center of Excellence in Fungal
Research, Thailand, are thanked for the valuable support provided for research.
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 11
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SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 15
SUPPLEMENTARY TABLE 1. Strains and related GenBank accession numbers of Diaporthe taxa used in this study.
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
Diaporthe acaciarum CBS 138862*KP004460 –KP004509 KP004504 –
D. acaciigena CBS 129521*KC343005 KC343731 KC343973 KC343489 KC343247
D. acericola MFLUCC 17-0956*KY964224 KY964180 KY964074 –KY964137
D. acerina CBS 137.27 KC343006 KC343732 KC343974 KC343490 KC343248
D. acuta PSCG 047*MK626957 MK654802 MK691225 MK726161 MK691124
D. acutispora CGMCC 3.18285*KX986764 KX999155 KX999195 KX999235 KX999274
D. albosinensis CFCC 53066*MK432659 MK578133 MK578059 MK443004 MK442979
D. alleghaniensis CBS 495.72*FJ889444 GQ250298 KC843228 KC343491 KC343249
D. alnea CBS 146.46*KC343008 KC343734 KC343976 KC343492 KC343250
D. ambigua CBS 114015*KC343010 KC343736 KC343978 KC343494 KC343252
D. ampelina CBS 114016*AF230751 GQ250351 JX275452 –JX197443
D. amygdali CBS 126679*KC343022 KC343748 KC343990 KC343506 KC343264
D. anacardii CBS 720.97*KC343024 KC343750 KC343992 KC343508 KC343266
D. angelicae CBS 111592*KC343026 KC343752 KC343994 KC343511 KC343268
D. anhuiensis CNUCC 201901 MN219718 MN224668 MN227008 MN224556 MN224549
D. apiculatum LC 3418*KP267896 KP267970 KP293476 ––
D. aquatica IFRDCC 3051*JQ797437 ––––
D. araucanorum RGM 2546 MN509711 MN509733 MN509722 –MN974277
D. arctii CBS 136.25 KC343031 KC343757 KC343999 KC343515 KC343273
D. arecae CBS 161.64*KC343032 KC343758 KC344000 KC343516 KC343274
D. arengae CBS 114979*KC343034 KC343760 KC344002 KC343518 KC343276
D. arezzoensis MFLU 19-2880*MT185503 MT454019 MT454055 ––
D. aseana MFLUCC 12-0299a*KT459414 KT459448 KT459432 –KT459464
D. asheicola CBS 136967*KJ160562 KJ160594 KJ160518 –KJ160542
D. aspalathi CBS 117169*KC343036 KC343762 KC344004 KC343520 KC343278
D. australafricana CBS 111886*KC343038 KC343764 KC344006 KC343522 KC343280
D. australiana BRIP 66145*MN708222 MN696522 MN696530
––
D. baccae CBS 136972*KJ160565 KJ160597 MF418509 MF418264 –
D. batatas CBS 122.21 KC343040 KC343766 KC344008 KC343524 KC343282
D. beckhausii CBS 138.27 KC343041 KC343767 KC344009 KC343525 KC343283
D. beilharziae BRIP 54792*JX862529 JX862535 KF170921 ––
D. benedicti CFCC 50062*KP208847 KP208853 KP208855 KP208851 KP208849
D. betulae CFCC 50469*KT732950 KT733016 KT733020 KT732999 KT732997
D. betulicola CFCC 51128*KX024653 KX024655 KX024657 KX024661 KX024659
D. bicincta CBS 121004*KC343134 KC343860 KC344102 KC343618 KC343376
D. biconispora CGMCC 3.17252*KJ490597 KJ490476 KJ490418 KJ490539 –
D. biguttulata ICMP20657*KJ490582 KJ490461 KJ490403 KJ490524 –
D. biguttusis CGMCC 3.17081*KF576282 KF576257 KF576306 ––
D. bohemiae CBS 143347*MG281015 MG281536 MG281188 MG281361 MG281710
D. brasiliensis CBS 133183*KC343042 KC343768 KC344010 KC343526 KC343284
D. caatingaensis CBS 141542*KY085927 KY115603 KY115600 KY115605 KY115597
D. camelliae-sinensis SAUCC194.92*MT822620 MT855932 MT855817 MT855588 MT855699
D. camptothecicola CFCC 51632*KY203726 KY228887 KY228893 KY228881 KY228877
D. canthii CBS 132533*JX069864 KC843120 KC843230 –KC843174
......continued on the next page
BUNDHUN ET AL.
16 • Phytotaxa 516 (1) © 2021 Magnolia Press
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. carpini CBS 114437 KC343044 KC343770 KC344012 KC343528 KC343286
D. cassines CBS 136440*KF777155 KF777244 –––
D. caulivora CBS 127268*KC343045 KC343771 KC344013 KC343529 KC343287
D. celastrina CBS 139.27*KC343047 KC343773 KC344015 KC343531 KC343289
D. celeris CBS 143349*MG281017 MG281538 MG281190 MG281363 MG281712
D. ceratozamiae CBS 131306*JQ044420 ––––
D. chamaeropis CBS 454.81 KC343048 KC343774 KC344016 KC343532 KC343290
D. charlesworthii BRIP 54884m*KJ197288 KJ197250 KJ197268 ––
D. chongqingensis PSCG435*MK626916 MK654866 MK691321 MK726257 MK691209
D. chromolaenae MFLUCC 17-1422*MT214362 ––––
D. chrysalidocarpi SAUCC194.35*MT822563 MT855876 MT855760 MT855532 MT855646
D. cichorii MFLUCC 17-1023*KY964220 KY964176 KY964104 –KY964133
D. cinerascens CBS 719.96 KC343050 KC343776 KC344018 KC343534 KC343292
D. cissampeli CBS 141331*KX228273 –KX228384 KX228366 –
D. citri CBS 135422*KC843311 KC843071 KC843187 MF418281 KC843157
D. citriasiana CBS 134240* JQ954645 JQ954663 KC357459 MF418282 KC357491
D. citrichinensis CBS 134242* JQ954648 JQ954666 MF418524 KJ420880 KC357494
D. compacta LC3083*KP267854 KP267928 KP293434 KP293508 –
D. convolvuli CBS 124654 KC343054 KC343780 KC344022 KC343538 KC343296
D. constrictospora CGMCC 3.20096*MT385947 MT424682 MT424702 MW022487 MT424718
D. coryli CFCC 53083*MK432661 MK578135 MK578061 MK443006 MK442981
D. crataegi CBS 114435 KC343055 KC343781 KC344023 KC343539 KC343297
D. crotalariae CBS 162.33*KC343056 KC343782 KC344024 KC343540 KC343298
D. crousii CAA823*MK792311 MK828081 MK837932 MK871450 MK883835
D. cucurbitae DAOM 42078*KM453210 KM453211 KP118848 KM453212 –
D. cuppatea CBS 117499*AY339322 AY339354 JX275420 KC343541 JX197414
D. cynaroidis CBS 122676 KC343058 KC343784 KC344026 KC343542 KC343300
D. cytosporella CBS 137020*KC843307 KC843116 KC843221 MF418283 KC843141
D. decedens CBS 109772 KC343059 KC343785 KC344027 KC343543 KC343301
D. decorticans CBS 114200 KC343169 KC343895 KC344137 KC343653 KC343411
D. delonicis MFLU 16-1059*MT215490 –MT212209 ––
D. destruens SPL15025*MH465671 MH560611 ––
MH560612
D. detrusa CBS 109770 KC343061 KC343787 KC344029 KC343545 KC343303
D. diospyricola CBS 136552*KF777156 ––––
D. discoidispora ICMP20662*KJ490624 KJ490503 KJ490445 KJ490566 –
D. drenthii BRIP 66524*MN708229 MN696526 MN696537 ––
D. durionigena KCSR1812.8*MN453530 MT276157 MT276159 ––
D. elaeagni-glabrae CGMCC 3.18287*KX986779 KX999171 KX999212 KX999251 KX999281
D. eleagni CBS 504.72 KC343064 KC343790 KC344032 KC343548 KC343306
D. ellipicola CGMCC 3.17084*KF576270 KF576245 KF576291 ––
D. ellipsospora CGMCC 3.20099*MT385949 MT424684 MT424704 MW022488 MT424720
D. endocitricola ZHKUCC 20-0012*MT355682 MT409336 MT409290 –MT409312
D. endophytica CBS 133811*KC343065 KC343791 KC344033 KC343549 KC343307
D. eres CBS 138594*KJ210529 KJ210550 KJ420799 KJ420850 KJ434999
......continued on the next page
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 17
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. eucalyptorum CBS 132525*JX069862 ––––
D. eugeniae CBS 444.82 KC343098 KC343824 KC344066 KC343582 KC343340
D. fibrosa CBS 109751 KC343099 KC343825 KC344067 KC343583 KC343341
D. foeniculina CBS 111553*KC343101 KC343827 KC344069 KC343585 KC343343
D. foikelawen RGM 2539*MN509713 MN509735 MN509724 –MN974278
D. forlicesenica MFLUCC 17-1015*KY964215 KY964171 KY964099 ––
D. forlicesenica MFLUCC 21-0011 MW677455 MW680161 MW680159 MW680160 –
D. fraxini-angustifoliae BRIP 54781*JX862528 JX852534 KF170920 ––
D. fructicola MAFF 246408*LC342734 LC342735 LC342736 LC342737 LC342738
D. fulvicolor PSCG 051*MK626859 MK654806 MK691236 MK726163 MK691132
D. fusicola CGMCC 3.17087*KF576281 KF576256 KF576305 –KF576233
D. ganjae CBS 180.91*KC343112 KC343838 KC344080 KC343596 KC343354
D. gardeniae CBS 288.56 KC343113 KC343839 KC344081 KC343597 KC343355
D. garethjonesii MFLUCC 12-0542a*KT459423 KT459457 KT459441 –KT459470
D. goulteri BRIP 55657a*KJ197290 KJ197252 KJ197270 ––
D. goulteri MFLUCC 21-0012 MW677456 MW680164 MW680162 MW680163 –
D. grandiflori SAUCC194.84*MT822612 MT855924 MT855809 MT855580 MT855691
D. guangdongensis ZHKUCC 20-0014*MT355684 MT409338 MT409292 –MT409314
D. guangxiensis JZB320094*MK335772 MK523566 MK500168 –MK736727
D. gulyae BRIP 54025*JF431299 JN645803 KJ197271 ––
D. guttulata CGMCC 3.20100*MT385950 MT424685 MT424705 MW022491 MW022470
D. heliconiae SAUCC194.77*MT822605 MT855917 MT855802 MT855573 MT855684
D. helianthi CBS 592.81*KC343115 KC343841 KC344083 KC343599 JX197454
D. heterophyllae CBS 143769*MG600222 MG600224 MG600226 MG600220 MG600218
D. heterostemmatis SAUCC194.85*MT822613 MT855925 MT855810 MT855581 MT855692
Diaporthe cf. heveae 1 CBS 852.97 KC343116 KC343842 KC344084 KC343600 KC343358
Diaporthe cf. heveae 2 CBS 681.84 KC343117 KC343843 KC344085 KC343601 KC343359
D. hickoriae CBS 145.26*KC343118 KC343844 KC344086 KC343602 KC343360
D. hispaniae CBS 143351*MG281123 MG281644 MG281296 MG281471 MG281820
D. hongkongensis CBS 115448*KC343119 KC343845 KC344087 KC343603 KC343361
D. hordei CBS 481.92 KC343120 KC343846 KC344088 KC343604 KC343362
D. huangshanensis CNUCC 201903*MN219729 MN224670 MN227010 MN224558 –
D. hubeiensis JZB320123* MK335809 MK523570 MK500148 –MK500235
D. humulicola CT2018-1 MN152927 MN180207 –MN180213 MN180204
D. hungariae CBS 143353*MG281126 MG281647 MG281299 MG281474 MG281823
D. impulsa CBS 114434 KC343121 KC343847 KC344089 KC343605 KC343363
D. incompleta CGMCC 3.18288*KX986794 KX999186 KX999226 KX999265 KX999289
D. inconspicua CBS 133813*KC343123 KC343849 KC344091 KC343607 KC343365
D. infecunda CBS 133812*KC343126 KC343852 KC344094 KC343610 KC343368
D. irregularis CGMCC 3.20092*MT385951 MT424686 MT424706 –MT424721
D. isoberliniae CBS 137981*KJ869133 –KJ869245 ––
D. italiana MFLUCC:18-0091*MH846238 MH853687 MH853689 –MH853691
D. juglandicola CFCC 51134*KU985101 KX024628 KX024634 –KX024616
D. kochmanii BRIP 54033*JF431295 JN645809 – – –
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BUNDHUN ET AL.
18 • Phytotaxa 516 (1) © 2021 Magnolia Press
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. kongii BRIP 54031*JF431301 JN645797 KJ197272 – –
D. krabiensis MFLUCC 17-2481*MN047100 MN433215 MN431495 ––
D. lenispora CGMCC 3.20101*MT385952 MT424687 MT424707 MW022493 MW022472
D. leucospermi CBS 111980*JN712460 KY435632 KY435673 KY435653 KY435663
D. limonicola CBS 142549*MF418422 MF418501 MF418582 MF418342 MF418256
D. litchicola BRIP 54900*JX862533 JX862539 KF170925 – –
D. litchii SAUCC194.22*MT822550 MT855863 MT855747 MT855519 MT855635
D. lithocarpus CGMCC 3.15175*KC153104 KC153095 KF576311 – –
D. longicicola CGMCC 3.17089*KF576267 KF576242 KF576291 – –
D. longicolla FAU 599*KJ590728 KJ590767 KJ610883 KJ659188 KJ612124
D. longispora CBS 194.36*KC343135 KC343861 KC344103 KC343619 KC343377
D. lonicerae MFLUCC 17-0963*KY964190 KY964146 KY964073 – KY964116
D. lusitanicae CBS 123212*KC343136 KC343862 KC344104 KC343620 KC343378
D. lutescens SAUCC194.36*MT822564 MT855877 MT855761 MT855533 MT855647
D. macadamiae BRIP 66526*MN708230 MN696528 MN696539 – –
D. machili SAUCC194.111*MT822639 MT855951 MT855836 MT855606 MT855718
D. macintoshii BRIP 55064a*KJ197289 KJ197251 KJ197269 – –
D. mahothocarpus CGMCC 3.15181*KC153096 KC153087 – – –
D. malorum CBS142383*KY435638 KY435627 KY435668 KY435648 KY435658
D. manihotia CBS 505.76 KC343138 KC343864 KC344106 KC343622 KC343380
D. marina MFLU 17-2622*MN047102 ––––
D. maritima DAOMC 250563*KU552025 KU552023 KU574615 – –
D. masirevicii BRIP 57892a*KJ197277 KJ197239 KJ197257 – –
D. mayteni CBS 133185*KC343139 KC343865 KC344107 KC343623 KC343381
D. maytenicola CBS 136441*KF777157 – KF777250 – –
D. mediterranea DAL-60 MT007492 MT006992 MT006689 MT007098 MT006764
D. megalospora CBS 143.27 KC343140 KC343866 KC344108 KC343624 KC343382
D. melastomatis SAUCC194.55*MT822583 MT855896 MT855780 MT855551 MT855664
D. melitensis CBS 142551*MF418424 MF418503 MF418584 MF418344 MF418258
D. melonis CBS 507.78*KC343142 KC343868 KC344110 KC343626 KC343384
D. middletonii BRIP 54884e*KJ197286 KJ197248 KJ197266 – –
D. millettiae GUCC9167*MK398674 MK480609 MK460488 –MK502086
D. minima CGMCC 3.20097*MT385953 MT424688 MT424708 MW022496 MT424722
D. minusculata CGMCC 3.20098*MT385957 MT424692 MT424712 MW022499 MW022475
D. miriciae BRIP 54736j*KJ197283 KJ197245 KJ197263 – –
D. momicola MFLUCC 16-0113*KU557563 KU557631 KU557587 – KU557611
D. multigutullata ICMP20656*KJ490633 KJ490512 KJ490454 KJ490575 –
D. musigena CBS 129519*KC343143 KC343869 KC344111 KC343627 KC343385
D. myracrodruonis URM 7972*NR_163320 MK213408 MK205291 –MK205290
D. neilliae CBS 144.27*KC343144 KC343870 KC344112 KC343628 KC343386
D. neoarctii CBS 109490 KC343145 KC343871 KC344113 KC343629 KC343387
D. nigra JZBH320170*MN653009 MN892277 MN887113 ––
D. nomurai CBS 157.29 KC343154 KC343880 KC344122 KC343638 KC343396
D. nothofagi BRIP 54801*JX862530 JX862536 KF170922 – –
......continued on the next page
SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 19
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. novem CBS 127271*KC343157 KC343883 KC344125 KC343641 KC343399
D. obtusifoliae CBS 143449*MG386072 – – MG386137 –
D. ocoteae CBS 141330*KX228293 – KX228388 – –
D. oncostoma CBS 589.78 KC343162 KC343888 KC344130 KC343646 KC343404
D. oraccinii LC 3166*KP267863 KP267937 KP293443 KP293517 –
D. osmanthi GUCC9165*MK398675 MK480610 MK502091 –MK502087
D. ovalispora ICMP20659*KJ490628 KJ490507 KJ490449 KJ490570 –
D. ovoicicola CGMCC 3.17092*KF576264 KF576239 KF576288 – KF576222
D. oxe CBS 133186*KC343164 KC343890 KC344132 KC343648 KC343406
D. paranensis CBS 133184 KC343171 KC343897 KC344139 KC343655 KC343413
D. parapterocarpi CBS 137986*KJ869138 – KJ869248 – –
D. parvae PSCG 034*MK626919 MK654858 MK691248 MK726210 –
D. patagonica RGM 2473*MN509717 MN509739 MN509728 –MN974279
D. pascoei BRIP 54847*JX862532 JX862538 KF170924 – –
D. passiflorae CBS 132527*JX069860 KY435633 KY435674 KY435654 KY435664
D. passifloricola CBS 141329*KX228292 – KX228387 KX228367 –
D. penetriteum LC 3353 KP714505 KP714517 KP714529 KP714493 –
D. perjuncta CBS 109745*KC343172 KC343898 KC344140 KC343656 KC343414
D. perniciosa CBS 124030 KC343149 KC343875 KC344117 KC343633 KC343391
D. perseae CBS 151.73 KC343173 KC343899 KC344141 KC343657 KC343415
D. pescicola MFLUCC 16-0105*KU557555 KU557623 KU557579 – KU557603
D. phaseolorum CBS 113425 KC343174 KC343900 KC344142 KC343658 KC343416
D. phillipsii MUM 19.28*MK792305 MK828076 MN000351 MK871445 MK883831
D. phragmitis CBS 138897*KP004445 – KP004507 KP004503 –
D. podocarpi-
macrophylli
CGMCC3.18281*KX986774 KX999167 KX999207 KX999246 KX999278
D. pometiae SAUCC194.72*MT822600 MT855912 MT855797 MT855568 MT855679
D. pseudomangiferae CBS 101339*KC343181 KC343907 KC344149 KC343665 KC343423
D. pseudophoenicicola CBS 462.69*KC343184 KC343910 KC344152 KC343668 KC343426
D. pseudotsugae MFLU 15-3228 KY964225 KY964181 KY964108 – KY964138
D. psoraleae CBS 136412*KF777158 KF777245 KF777251 – –
D. psoraleae-pinnatae CBS 136413*KF777159 – KF777252 – –
D. pterocarpi MFLUCC 10-0571 JQ619899 JX275416 JX275460 – JX197451
D. pterocarpicola MFLUCC 10-0580a JQ619887 JX275403 JX275441 – JX197433
D. pungensis SAUCC194.112*MT822640 MT855952 MT855837 MT855607 MT855719
D. pustulata CBS 109742 KC343185 KC343911 KC344153 KC343669 KC343427
D. pyracanthae CBS 142384*KY435635 KY435625 KY435666 KY435645 KY435656
D. racemosae CBS 143770*MG600223 MG600225 MG600227 MG600221 MG600219
D. raonikayaporum CBS 133182*KC343188 KC343914 KC344156 KC343672 KC343430
D. ravennica MFLUCC 15-0479*KU900335 – KX432254 – –
D. rhoina CBS 146.27 KC343189 KC343915 KC344157 KC343673 KC343431
D. rhusicola MFLUCC 16-1393 KY684947 KY684946 KY684945 – –
D. rosiphthora COAD 2914*MT311197 – – – –
D. rossmaniae MUM 19.30*MK792290 MK828063 MK837914 MK871432 MK883822
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BUNDHUN ET AL.
20 • Phytotaxa 516 (1) © 2021 Magnolia Press
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. rostrata CFCC 50062*KP208847 KP208853 KP208855 KP208851 KP208849
D. rudis CBS 113201*KC343234 KC343960 KC344202 KC343718 KC343476
D. saccarata CBS 116311*KC343190 KC343916 KC344158 KC343674 KC343432
D. sackstonii BRIP 54669b*KJ197287 KJ197249 KJ197267 – –
D. salicicola BRIP 54825*JX862531 JX862537 KF170923 – –
D. salinicola MFLUCC 18-0553 *MN047098 MN077073 – – –
D. sambucusii CFCC 51986*KY852495 KY852507 KY852511 KY852503 KY852499
D. schini CBS 133181*KC343191 KC343917 KC344159 KC343675 KC343433
D. schisandrae CFCC 51988*KY852497 KY852509 KY852513 KY852505 KY852501
D. schoeni MFLU 15-1279*KY964226 KY964182 KY964109 – KY964139
D. sclerotioides CBS 296.67*KC343193 KC343919 KC344161 KC343677 KC343435
D. scobina CBS 251.38 KC343195 KC343921 KC344163 KC343679 KC343437
D. searlei BRIP 66528*MN708231 – MN696540 – –
D. sennae CFCC 51636*KY203724 KY228885 KY228891 – KY228875
D. sennicola CFCC 51634*KY203722 KY228883 KY228889 – KY228873
D. serafiniae BRIP 55665a*KJ197274 KJ197236 KJ197254 – –
D. shaanxiensis CFCC 53106*MK432654 MK578130 –MK443001 MK442976
D. shennongjiaensis CNUCC 201905*MN216229 MN224672 MN227012 MN224560 MN224551
D. siamensis MFLUCC 10-0573a JQ619879 JX275393 JX275429 – –
D. sojae CBS 139282*KJ590719 KJ590762 KJ610875 KJ659208 KJ612116
D. spinosa PSCG383*MK626849 MK654811 MK691234 MK726156 MK691129
D. sterilis CBS 136969*KJ160579 KJ160611 KJ160528 MF418350 KJ160548
D. stewartii CBS 193.36 FJ889448 GQ250324 – – –
D. stictica CBS 370.54 KC343212 KC343938 KC344180 KC343696 KC343454
D. subclavata ICMP20663*KJ490630 KJ490509 KJ490451 KJ490572 –
D. subordinaria CBS 101711 KC343213 KC343939 KC344181 KC343697 KC343455
D. taoicola MFLUCC 16-0117*KU557567 KU557635 KU557591 – –
D. tarchonanthi CPC 37479*MT223794 –MT223733 MT223759 –
D. tecomae CBS 100547 KC343215 KC343941 KC344183 KC343699 KC343457
D. tectonae MFLUCC 12-0777*KU712430 KU749359 KU743977 – KU749345
D. tectonendophytica MFLUCC 13-0471*KU712439 KU749367 KU743986 – KU749354
D. tectonigena MFLUCC 12-0767*KU712429 KU749371 KU743976 – KU749358
D. terebinthifolii CBS 133180*KC343216 KC343942 KC344184 KC343700 KC343458
D. ternstroemia CGMCC 3.15183*KC153098 KC153089 – – –
D. thunbergii MFLUCC 10-0756a JQ619893 JX275409 JX275449 – JX197440
D. toxicodendri FFPRI420987 LC275192 LC275216 LC275224 LC275216 LC275200
D. tulliensis BRIP 62248a KR936130 KR936133 KR936132 – –
D. ueckerae FAU 656 KJ590726 KJ590747 KJ610881 KJ659215 KJ612122
D. undulata CGMCC 3.18293*KX986798 KX999190 KX999230 KX999269 –
D. unshiuensis CGMCC3.17569*KJ490587 KJ490466 KJ490408 KJ490529 –
D. vaccinii CBS 160.32*AF317578 GQ250326 KC344196 KC343712 KC343470
D. vacuae MUM 19.31*MK792309 MK828080 MK837931 MK871449 MK883834
D. vangueriae CBS 137985*KJ869137 – KJ869247 – –
D. vawdreyi BRIP 57887a KR936126 KR936129 KR936128 –
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SEXUAL MORPHS OF DIAPORTHE FORLICESENICA Phytotaxa 516 (1) © 2021 Magnolia Press • 21
SUPPLEMENTARY TABLE 1. (Continued)
Species Strain GenBank accession
numbers
ITS tef1 tub2 his cal
D. velutina CGMCC 3.18286*KX986790 KX999182 KX999223 KX999261 –
D. vexans CBS 127.14 KC343229 KC343955 KC344197 KC343713 KC343471
D. viniferae JZB320071*MK341550 MK500107 MK500112 –MK500119
D. virgiliae CBS 138788*KP247573 – KP247582 – –
D. woodii CBS 558.93 KC343244 KC343970 KC344212 KC343728 KC343486
D. woolworthii CBS 148.27 KC343245 KC343971 KC344213 KC343729 KC343487
D. xishuangbanica CGMCC 3.18282*KX986783 KX999175 KX999216 KX999255 –
D. yunnanensis CGMCC 3.18289*KX986796 KX999188 KX999228 KX999267 KX999290
D. zaobaisu PSCG031*MK626922 MK654855 MK691245 MK726207 –
Diaporthella corylina CBS 121124*KC343004 KC343730 KC343972 KC343488 KC343246
Ex-types are indicated with *; newly generated sequences in this study are in bold blue.
Abbreviations: BRIP—Queensland Plant Pathology Herbarium, Brisbane, Australia; CBS—Westerdijk Fungal Biodiversity Institute,
Utrecht, the Netherlands; CFCC—China Forestry Culture Collection Center, Beijing, China; CGMCC—Chinese General Microbiological
Culture Collection Center, Beijing, China; CNUCC—Capital Normal University Culture Collection Center, Beijing, China; DAOM—
Plant Research Institute, Department of Agriculture (Mycology), Ottawa, Canada; DAOMC—Canadian Collection of Fungal Cultures,
Ottawa, Canada; FAU—Isolates in culture collection of Systematic Mycology and Microbiology Laboratory; FFPRI—Forestry and Forest
Products Research Institute, Japan; ICMP—International Collection of Micro-organisms from Plants, Landcare Research, Private Bag
92170, Auckland, New Zealand; IFRDCC—International Fungal Research and Development Culture Collection; JZB—Culture collection
of Institute of Plant and Environment Protection of Beijing Academy of Agriculture and Forestry Sciences; LC— working collection of Lei
Cai, housed at Institute of Microbiology, Chinese Academy of Sciences, Beijing, China; MFLU—Mae Fah Luang University herbarium,
Thailand; MFLUCC—Mae Fah Luang University Culture Collection, Chiang Rai, Thailand; MUM—Fungal culture collection from
Micoteca da Universidade do Minho, Portugal; SAUCC— Shandong Agricultural University Culture Collection. ZHKUCC—Zhongkai
University Culture Collection.
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SUPPLEMENTARY FIGURE 1. RAxML tree based on analysis of a combined dataset of ITS, tef1, tub2, his and cal sequence data.
Bootstrap support values for ML values ≥ 50% and BYPP values ≥ 0.90 are shown as ML/BYPP above or below the branches respectively.
The isolates used for the study are indicated in bold red while the ex-types, in bold black. The tree is rooted with Diaporthella corylina
(CBS 121124). The scale bar represents the expected number of nucleotide substitutions per site. The best scoring RAxML tree with final
optimization had a likelihood value of -91990.044067. The matrix had 2,578 distinct alignment patterns, with 40.40% of undetermined
characters or gaps. Estimated base frequencies were as follows: A= 0.211725, C= 0.330857, G= 0.235547, T= 0.221871; substitution rates
AC= 1.089796, AG= 3.300330, AT= 1.110042, CG= 0.855613, CT= 4.308347, GT= 1.000000; gamma distribution shape parameter α=
0.505465; Tree-length= 14.763726.
