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Ficus microcarpa (Moraceae) and Kandelia candel (Rhizophoraceae), Two New

Host Records for Alternaria gaisen from Taiwan

Article  in  Chiang Mai Journal of Science · November 2021

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 Chiang Mai J. Sci. 2021; 48(6) : 1478-1499
http://epg.science.cmu.ac.th/ejournal/
Contributed Paper

Ficus microcarpa (Moraceae) and Kandelia candel

(Rhizophoraceae), Two New Host Records for Alternaria
gaisen from Taiwan
Chada Norphanphoun [a,b,d,e], Sinang Hongsanan [f], Jayarama Darbhe Bhat [c], Chang-Hsin Kuo*
[a] and Kevin David Hyde [b,d,g]
[a] Department of Plant Medicine, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, Taiwan
[b] Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand
[c] House No. 128/1-J, Azad Housing Society, Curca, Goa Velha 403108, India
[d] Mushroom Research Foundation, 128 M.3 Ban Pa Deng T. Pa Pae, A. Mae Taeng, Chiang Mai 50150, Thailand
[e] School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
[f] Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen Key Laboratory of Microbial Genetic
Engineering, College of Life Science and Oceanography, Shenzhen University, Shenzhen 518060, People’s
Republic of China
[g] Innovative Institute of Plant Health, Zhongkai University of Agriculture and Engineering, Haizhu District, Guangzhou
510225, P.R. China
*Author for correspondence; e-mail: chkuo@mail.ncyu.edu.tw
Received: 31 January 2021
Revised: 25 May 2021
Accepted: 1 June 2021
ABSTRACT
Mangroves grow adjacent to estuarine regions of the tropics and subtropics. Fungi cause
diseases in mangroves; however, it is not clear if the diseases are unique to mangrove species or
ubiquitous. In this study, we identified a species of Alternaria, a genus comprising with saprobic and
pathogenic species of a large number of plants. The genus is currently divided into 28 sections based on
phylogenetic and morphological evidence. Alternaria is characterized by macronematous conidiophores,
sympodial, polytretic conidiogenous cells, and obclavate to obovoid, muriform conidia gradually
tapering towards beaked apex. We found A. gaisen associated with leaf blight of Ficus microcarpa and
leaf spots of Kandelia candel from Taiwan. Phylogenetic analysis of combined SSU, LSU, ITS, gapdh,
rpb2, ef1α, Alta1, endoPG and OPA10-2 sequence data supports positioning of our isolates as A. gaisen
in section Alternaria. This species was first reported from Japanese pear causing black spot disease.
This is also a new host record of A. gaisen from mangroves in Taiwan.

Keywords: new host record, mangroves, microfungi, phylogenetic analyses, pleosporaceae, Alternaria
gaisen

1. INTRODUCTION
Alternaria Nees is a genus composed of many
important plant pathogens as well as endophytic
and saprobic species with a worldwide distribution
[5], [59]. Species of Alternaria mainly inflict damage
to pre- and post-harvest agricultural products
[37]. Some Alternaria species produce secondary
metabolites considered as phytotoxins (harmful
to plants), and mycotoxins (harmful to humans
and animals) [37].
Chiang Mai J. Sci. 2021; 48(6) 1479
Alternaria was introduced by Nees von
Esenbeck [31] to accommodate the type species,
A. alternata (Fr.) Keissl. (=A. tenuis Nees). It is
currently placed in the family Pleosporaceae,
Pleosporales, Dothideomycetes, based on taxonomy
and phylogenetic evidence [18], [57], [58]. Alternaria
species produce concatenated conidia, typically
ovoid to obclavate, observed singly or in chains,
darkly pigmented, muriform, with the basal end
of the conidium being rounded, while it tapers
towards the apex and gives a typical beak or
club-like appearance of the conidium [26]. Over
700 species of this genus are listed in Index
Fungorum (accessed January 23, 2021). The
classification of Alternaria has been confusing
in most cases and so far conidial size and shape
were used to determine the distinction between
different species. Phylogenetic analyses indicated
that, there are 28 internal clades of the Alternaria
complex i.e. sect. Alternantherae, sect. Alternaria,
sect. Brassicicola, sect. Chalastospora, sect. Cheiranthus,
sect. Crivellia, sect. Dianthicola, sect. Embellisia,
sect. Embellisioides, sect. Euphorbiicola, sect. Eureka,
sect. Gypsophilae, sect. Infectoriae, sect. Japonicae,
sect. Nimbya, sect. Omanenses, sect. Panax, sect.
Phragmosporae, sect. Porri, sect. Pseudoalternaria,
sect. Pseudoulocladium, sect. Radicina, sect. Soda,
sect. Sonchi, sect. Teretispora, sect. Ulocladioides, sect.
Ulocladium, and sect. Undifilum [2], [25-27], [59].
The sect. Alternaria was formalized to a
section by Lawrence et al. [25], with A. alternata
(Fr.) Keissl. as the type species, while seven
sections were introduced in the same study.
Section Alternaria includes many important species
causing diseases on economic plants, as well as
post-harvest pathogens, and human pathogens, viz.
A. arborescens Simmons, A. alternata (Fr.) Keissl.,
A. gaisen Nagano ex Hara, A. longipes, A. mali
(Ellis & Everh.) Mason, and A. tenuissima (Kunze)
Wiltshire [45], [60]. This section is characterized by
transversely and longitudinally septate, obclavate to
ellipsoidal conidia, produced on simple or branched
conidiophores, with some forming conidial chains
and bearing one or several tretic conidiogenous
cells (with sympodial proliferations) [25], [59].
Mangroves occur in estuarine regions in the
tropics and subtropics [19] and suffer from diseases
cause by fungi [7], [14], [20], [23], [28], [39], [41],
[44], [48], [53], [61]. This paper introduces a new
host record of Alternaria gaisen, associated with
leaf blight of Ficus microcarpa L.f. (Moraceae),
plant species that occur mainly at low elevations
and natural habitats including tropical rainforests,
river edges, coasts, swamps, and mangrove areas
[13]; and leaf spots of Kandelia candel (L.) Druce
(Rhizophoraceae), an abundant mangrove plant in
Taiwan that occurs around the coasts and saline-
brackish habitats of South Asia and Southeast
Asia, which was planted in order to check winds,
control sand and protect dikes [43].
2. MATERIAL AND METHODS
2.1 Sample Collection and Specimen Examination
Fresh samples with leaf blight and leaf-spot
symptoms were collected in July, 2018, from Ficus
microcarpa and Kandelia candel. Fresh specimens were
taken to the laboratory in paper bags, examined
and described. Mega-morphological characters
were examined using a Zeiss Axioskop 2 plus
stereo microscope (Carl Zeiss Microscopy, LLC,
NY, USA). Micro-morphology was examined
using a Zeiss Axioskop 2 mot plus compound
microscope (Carl Zeiss Microscopy, LLC, NY,
USA). Images’ measurements were made using
ZEN2 (blue edition) software. Adobe Photoshop
CC 2019 version: 20.0.1 was used to prepare fungal
photo-plates (Adobe Systems, CA, USA) [42].
Pure cultures were obtained by single spore
isolation for fungi from Ficus microcarpa. A conidial
suspension was prepared by scraping spore masses
from the surface of substrate using a sterilized
needle into a drop of sterile water on a small glass
container. The conidial suspension was examined
using a stereo microscope, sucked by a Pasteur
pipette or syringe and placed in drops on 2% water
agar (WA) at the center of squares marked in a
grid on the bottom of Petri dish and incubated at
room temperature (25 °C ± 2) overnight. Single
1480 Chiang Mai J. Sci. 2021; 48(6)
germinating spores observed using a dissecting
microscope were transferred aseptically into 2%
potato dextrose agar (PDA) and incubated at room
temperature for one-week under a daily light/dark
cycle. Tissue isolation method of Norphanphoun
et al. [34] was used to isolate the fungus from leaf
spot of Kandelia candel. Based on Simmons [47],
fungal isolations in this study were cultured into
PDA, potato-carrot agar (PCA), and V-8 juice
agar (V-8 agar) Petri-dish and incubated at room
temperature for one-week period under a daily
light/dark cycle. Culture characters were examined
and recorded at intervals of 5, 7, 14 and 30 d.
Morphological characters of fungi on media were
examined, r+ecorded, and compared with other
species published based on Alternaria species
keying system in Simmons [47]. Pure cultures
were maintained for further studies in PDA, PCA
and V-8 agar. The dried and living cultures are
deposited in the Department of Plant Medicine,
National Chiayi University (NCYU) and duplicates
in culture collection Mae Fah Luang University
(MFLUCC), Chiang Rai, Thailand.
2.2 DNA Extraction, PCR Amplification and
Sequencing
DNA extraction was performed with
fresh fungal mycelia growing on PDA at room
temperature (25 °C ± 2) for one week using a
E.Z.N.A ® Fungal DNA Mini Kit, D3390-02,
(Omega Bio-tek, Inc., GA) and following the
manufacturer’s protocols. The small subunit rRNA
(SSU) was amplified using primer pairs NS1/NS4
[56]; the 28S large subunit ribosomal RNA gene
(LSU) amplified by primers LROR/LR5, [56];
the internal transcribed spacer region (ITS) using
primers ITS1/ ITS4 [56]; translation elongation
factor 1-α (ef1α) using primers 728f/986r [10];
glyceraldehyde-3-phosphate dehydrogenase (gapdh)
using primers gpd1/gpd2 [9]; RNA polymerase
II subunit (rpb2) using primers 5f2/7cr [59];
Alternaria major allergen (Alta1) using primers
Alt-for/ Alt-rev [17]; the endopolygalacturonase
(endoPG) was amplified using the primers PG3 and
PG2b; and an anonymous gene region (OPA10-2)
was amplified using the primers OPA10-2L and
OPA10-2R [4].
The polymerase chain reactions (PCR) were
carried out with 50 µL reaction volume (2 µL of
DNA template, 2 µL of each forward and reverse
primers, 25 µL of 2×Power Taq PCR MasterMix
(Tri-I Biotech, Taipei, Taiwan) and 19 µL of
double-distilled water (ddH2O)). The PCR thermal
cycle programs was carried out for each gene as,
SSU: 95 °C (3 min), 40 cycles of 95 °C (30 s),
52 °C (50 s), 72 °C (1 min), final extension at 72 °C
(7 min); LSU: 95 °C (3 min), 34 cycles of 95 °C
(30 s), 51 °C (50 s), 72 °C (1 min), final extension
at 72 °C (10 min); ITS: 95 °C (3 min), 40 cycles
of 95 °C (30 s), 55 °C (50 s), 72 °C (1 min), final
extension at 72 °C (7 min); ef1α: 94 °C (3 min),
40 cycles of 94 °C (45 s), 52 °C (30 s), 72 °C
(30 s), final extension at 72 °C (6 min). gapdh PCR
thermal cycle programs was followed Al Ghafri
et al. [2]; rpb2 was followed from Woudenberg
et al. [59]; Alt a 1 was followed from Hong et
al. [17]; endoPG was followed from Andrew et al.
[4] and Woudenberg et al. [60]. OPA10-2: 94 °C
(5 min), 35 cycles of 94 °C (30 s), 50 °C (30 s),
72 °C (45 s), final extension at 72 °C (10 min).
PCR products were analyzed with 1.5% agarose gel
containing the Safeview DNA stain (GeneMark,
Taipei, Taiwan). Purification and sequencing were
carried out at Tri-I Biotech, Taipei, Taiwan.
2.3 Phylogenetic Analysis
Geneious® 11.1.5 (http://www.geneious.
com) was used to assemble sequence data. Multiple
alignments derived in this study were analyzed
with similar sequences, acquired from GenBank
BLASTn included recently published [2], [60]. ITSx
1.1 a Perl based software tool was used to extract
SSU, ITS1, 5.8S, ITS2 and LSU [8]. BioEdit 7.2.3
[16] was used to extract the partition of gapdh,
ef1α, and Alta1 based on nucleotide BLAST in
GenBank. Combined sequence data of Figure 1
(SSU, LSU, ITS1+ ITS2, 5.8S, gapdh-exon, rpb2,
ef1α-exon, Alta1-exon, gapdh-intron, ef1α-intron,
Chiang Mai J. Sci. 2021; 48(6) 1481

Figure 1. Phylogram generated from maximum likelihood analysis based on SSU, LSU, ITS1+ ITS2,
5.8S, gapdh-exon, rpb2, ef1α-exon, Alta1-exon, gapdh-intron, ef1α-intron, and Alta1-intron sequence data
of 28 Alternaria sections (97 Alternaria strains). The tree is rooted to Stemphylium herbarum strain CBS
191.86 and Pleospora tarda strain CBS 714.68. Maximum parsimony and maximum likelihood bootstrap
values ≥50%, Bayesian posterior probabilities ≥0.90 (MPBS/MLBS/BIPP) are given at the nodes.
The species obtained in this study are in blue. Ex-type strains are in bold.
1482 Chiang Mai J. Sci. 2021; 48(6)
and Alta1-intron) and Figure 2 (SSU, LSU,
ITS1+ITS2, 5.8S, gapdh-exon, rpb2, ef1α-exon,
Alta1-exon, endoPG, OPA10-2, gapdh-intron, ef1α-
intron, and Alta1-intron) were performed with
maximum parsimony (MP), maximum likelihood
(ML) and Bayesian analysis (BI). NEXUS file was
converted by using Geneious Prime® 2019.2.1
(http://www.geneious.com). MrModeltest v. 2.2
[35] was prepared in PAUP* v.4.0b10 [50]. Dataset
of phylogenetic tree in Figure 1 consisted 99 taxa,
included outgroup strains (Stemphylium herbarum
strain CBS 191.86 and Pleospora tarda strain
CBS 714.68). This tree was used to determine
all Alternaria sections (28 sections). Dataset of
Figure 2 consisted 41 taxa of Alternaria sect.
Alternaria and outgroup from Alternaria sect.
Alternantherae (A. perpunctulata strain CBS 115267
and A. alternantherae strain CBS124392). Ingroup
taxa were selected based on preliminary analyses
(not shown) and Figure 1. Alignments of each
gene region were aligned in MAFFT v.7.110
online program (http://mafft.cbrc.jp/alignment/
server/; [22]); Phylemon 2.0 were used to exclude
ambiguously aligned positions alignments [40] and
manual adjustments were made wherever necessary
in BioEdit v. 7.0.5.3 [16]. MEGA v. 10.1.0 was
used to combine sequence [24]. Missing data were
coded as Question mark (?). The phylogram was
visualized in FigTree v1.4.0 (http://tree.bio.ed.ac.
uk/software /figtree/; [38]) and performed by
using Adobe Illustrator CC version 23.0.1 (Adobe
Systems, CA, USA). Newly generated sequences
in this study are deposited in GenBank (Table 1).
The finalized alignments and tree topology were
deposited in TreeBASE, Figure 1: submission ID:
26854, Figure 2: submission ID: 26855.
Maximum parsimony (MP), Maximum likelihood
(ML) and Bayesian inference (BI) analyses were
partitioned analysis and performed for Figure 1
with 11 partitions: SSU, LSU, ITS1+ITS2, 5.8S,
gapdh-exon, rpb2, ef1α-exon, Alta1-exon, gapdh-intron,
ef1α-intron, and Alta1-intron) and for Figure 2
with 13 partitions: SSU, LSU, ITS1+ITS2, 5.8S,
gapdh-exon, rpb2, ef1α-exon, Alta1-exon, endoPG,
OPA10-2, gapdh-intron, ef1α-intron, and Alta1-
intron. Analyses were implemented in CIPRES
Science Gateway web server with tool selected,
PAUP on XSEDE, RAxML-HPC2on XSEDE,
and MrBayes on XSEDE respectively [29]. 1000
bootstrap replicates using heuristic search with
random stepwise addition and tree-bisection
reconnection (TBR) for Maximum parsimony.
1000 rapid bootstrap replicates were run with
GTRGAMMA model of nucleotide evolution
was set for Maximum likelihood and bootstrap
values for MP and ML were equal or greater than
50% are presented in both trees.
Bayesian inference (BI) analysis was performed
using the Markov Chain Monte Carlo (MCMC)
with best-fit nucleotide substitution model for
each gene and dataset were determined using
MrModel test version 2.2 with selected by AIC
[35]. In Figure 1: HKY+I was selected as a best-fit
model for SSU; GTR+I+G was selected for LSU,
ITS1+ITS2, gapdh-exon, and rpb2; JC for 5.8S; K80
for ef1α-exon; HKY+G for Alta1-exon; HKY+I+G
for gapdh-intron, and ef1α-intron; and GTR+I was
selected as a best-fit model for Alta1-intron. In
Figure 2: HKY was selected as a best-fit model
for SSU, ef1α-intron, and Alta1-intron; HKY+I
for LSU, Alta1-exon, and gapdh-intron; K80 for
ITS1+ITS2, 5.8S, and ef1α-exon; GTR+I for
gapdh-exon; SYM+G for rpb2; SYM+I for endoPG;
and K80+G for OPA10-2;The MCMC analyses
and four chains random tree topology were run
between 10,000,000 and 1,000,000 generations
respectively for each combined dataset. Trees
were sampled every 100 generations with 25.0%
of relative burn-in value.
3. RESULTS
3.1 Phylogenetic Analysis
The phylogenetic tree in Figure 1 included
99 strains with Stemphylium herbarum (CBS 191.86)
and Pleospora tarda (CBS 714.68) as outgroup. The
total number of characters in the dataset was 4352,
including alignment gaps, with 1–1010, 1011–1847,
1848–2171, 2172–2329, 2330–2740, 2741–3475,
Chiang Mai J. Sci. 2021; 48(6) 1483

Figure 2. Phylogram generated from maximum likelihood analysis based on combined SSU, LSU,
ITS1+ ITS2, 5.8S, gapdh-exon, rpb2, ef1α-exon, Alta1-exon, endoPG, OPA10-2, gapdh-intron, ef1α-intron
, and Alta1-intron,sequence data for 41 Alternaria stains in section Alternaria. The tree is rooted to
two Alternaria stains in section Alternantherae (A. perpunctulata strain CBS 115267 and A. alternantherae
strain CBS 124392). Maximum parsimony and maximum likelihood bootstrap values ≥50%, Bayesian
posterior probabilities ≥0.90 (MPBS/MLBS/BIPP) are given at the nodes. The species obtained in
this study are in blue. Ex-type strains are in bold.
Table 1. GenBank accession numbers of the sequences used in this study.
GenBank accession numbers
1484

Species Strain * Location Host

SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
T
Alternaria abundans CBS 534.83 New Zealand Fragaria KC584581 KC584323 JN383485 KC584154 KC584448 KC584707 JN383503 - -
A. alstroemeriae CBS 118808 USA Alstroemeria sp. KP124917 KP124447 KP124296 KP124153 KP124764 KP125071 KP123845 KP123993 KP124601
T
A. alstroemeriae CBS 118809 Australia Alstroemeria sp. KP124918 KP124448 KP124297 KP124154 KP124765 KP125072 - KP123994 KP124602
A. alternantherae CBS 124392 China Solanum melongena KC584506 KC584251 KC584179 KC584096 KC584374 KC584633 KP123846 - -
A. alternariae CBS 126989 USA Daucus carota KC584604 KC584346 AF229485 - KC584470 KC584730 - - -
A. alternata CBS 117.44 Denmark Godetia sp. KP124925 KP124455 KP124303 KP124160 KP124772 KP125079 KP123854 KP124001 KP124609
A. alternata CBS 174.52 USA Anemone occidentalis KC584578 DQ678068 KC584228 KC584152 DQ677964 KC584704 KP123856 KP124003 KP124611
A. alternata CBS 686.68 Sahara desert sand KP124928 KP124458 KP124306 KP124163 KP124775 KP125082 KP123859 KP124006 KP124614
A. alternata CBS 826.68 Germany Lolium sp. KP124929 KP124459 KP124307 KP124164 KP124776 KP125083 KP123860 KP124007  
T
A. alternata CBS 916.96 India Arachis hypogaea KC584507 DQ678082 KF465761 - KC584375 KC584634 AY563301 JQ811978 KP124632
A. alternata CBS 102595 USA Citrus jambhiri KC584540 KC584284 FJ266476 AY562411 KC584408 KC584666 AY563306 KP124029 KP124636
A. alternata CBS 116749 Netherlands unknown KP124975 KP124505 KP124353 KP124207 KP124821 KP125129 KP123901 KP124054 KP124664
A. alternata CBS 118812 USA Daucus carota KC584525 KC584269 KC584193 KC584112 KC584393 KC584652 KP123905 KP124058 KP124668
A. arborescens CBS 101.13 Switzerland peat soil KP125016 KP124546 KP124392 KP124244 KP124862 KP125170 KP123940 KP124096 KP124705
T
A. arborescens CBS 102605 USA Solanum lycopersicum KC584509 KC584253 NR_135927 AY278810 KC584377 KC584636 AY563303 AY295028 KP124712
A. arborescens CPC 4244 South Africa Malus domestica KP125023 KP124553 KP124400 KP124252 KP124870 KP125178 KP123947 KP124104 KP124714
A. arborescens CBS 119544 New Zealand Avena sativa KP125031 KP124561 KP124408 JQ646321 KP124878 KP125186 KP123955 KP124112 KP124722
A. arborescens CBS 123235 Denmark human toenail KP125033 KP124563 KP124410 KP124261 KP124880 KP125188 KP123957 KP124114 KP124724
A. arborescens CBS 124283 Russia Oryza sp. KP125039 KP124569 KP124416 KP124267 KP124885 KP125194 KP123963 KP124120 KP124730
A. argyranthemi CBS 116530 New Zealand Argyranthemum sp. KC584510 KC584254 KC584181 KC584098 KC584378 KC584637 - - -
A. arrhenatheri BMP 0514  -  - - - JQ693680 JQ693629 - - - - -
T
A. aspera CBS 115269 Japan Pistacia vera KC584607 KC584349 KC584242 KC584166 KC584474 KC584734 KF533899 - -
T
A. axiaeriisporifera CBS 118715 New Zealand Gypsophila paniculata KC584513 KC584257 KC584184 KC584101 KC584381 KC584640 - - -
T
ex-type strain, strain in this study are in bold.
Chiang Mai J. Sci. 2021; 48(6)
Table 1. (Continued).
GenBank accession numbers
Species Strain * Location Host
SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
T
A. betae-kenyensis CBS 118810 Kenya Beta vulgaris var. cicla KP125042 KP124572 KP124419 KP124270 KP124888 KP125197 KP123966 KP124123 KP124733
A. bornmueller DAOM 231361 Austria Securigera varia KC584624 KC584366 FJ357317 FJ357305 KC584491 KC584751 JN383516 - -
T
A. botryospora CBS 478.90 New Zealand Leptinella dioica KC584594 KC584336 MH862228 AY278831 KC584461 KC584720 - - -
T
A. botrytis CBS 197.67 USA  - KC584609 KC584351 KC584243 KC584168 KC584476 KC584736 - - -
A. brassicae CBS 116528 USA Brassica oleracea KC584514 KC584258 KC584185 KC584102 KC584382 KC584641 - - -
Chiang Mai J. Sci. 2021; 48(6)

T Brassica rapa subsp.

A. brassicae-pekinensis CBS 121493 China KC584611 KC584353 KC584244 KC584170 KC584478 KC584738 KF533892 - -
Pekinensis
A. brassicicola CBS 118699 USA Brassica oleracea KC584515 KC584259 JX499031 KC584103 KC584383 KC584642 - - -
T
A. breviramosa CBS 121331 Australia Triticum sp. KC584574 KC584318 FJ839608 KC584148 KC584442 KC584700 - - -
T
A. burnsii CBS 107.38 India Cuminum cyminum KP125043 KP124573 KP124420 JQ646305 KP124889 KP125198 KP123967 KP124124 KP124734
A. burnsii CBS 110.50 Mozambique Gossypium sp. KP125044 KP124574 KP124421 KP124271 KP124890 KP125199 KP123968 KP124125 KP124735
T
A. calycipyricola CBS 121545 China  - KC584516 KC584260 KC584186 KC584104 KC584384 KC584643 - - -
A. capsici-annui CBS 504.74  - Capsicum annuum KC584517 KC584261 KC584187 KC584105 KC584385 KC584644 - - -
T
A. caricis CBS 480.90 USA Carex hoodii NG_062917 NG_057022 NR_135934 AY278826 KC584467 KC584726 - - -
T
A. carotiincultae CBS 109381 USA Daucus carota KC584518 KC584262 KC584188 KC584106 KC584386 KC584645 - - -
A. cetera CBS 121340 T Australia Elymus scabrus KC584573 KC584317 FJ839607 AY562398 KC584441 KC584699 - - -
T
A. chartarum CBS 200.67 Canada Populus sp. KC584614 KC584356 AF229488 KC584172 KC584481 KC584741 - - -
A. cheiranthi CBS 109384 Italy Erysimum cheiri KC584519 KC584263 AF229457 KC584107 KC584387 KC584646 JQ905106 - -
T
A. chlamydospora CBS 491.72 Egypt soil KC584520 KC584264 KC584189 KC584108 KC584388 KC584647 JQ646367 - -
A. chlamydosporigena CBS 341.71 USA air KC584584 KC584326 KC584231 KC584156 KC584451 KC584710 - - -
A. cinerariae CBS 116495 USA Ligularia sp. KC584521 KC584265 KC584190 KC584109 KC584389 KC584648 - - -
T
A. concatenata CBS 120006  -  - KC584613 KC584355 KC584246 - KC584480 KC584740 - - -
T
A. conjuncta CBS 196.86 Switzerland Pastinaca sativa KC584522 KC584266 FJ266475 AY562401 KC584390 KC584649 - - -
A. conoidea CBS 132.89 Saudi Arabia Ricinus communis KC584585 KC584327 FJ348226 FJ348227 KC584452 KC584711 FJ348228 - -
T
ex-type strain, strain in this study are in bold.
1485
Table 1. (Continued).
GenBank accession numbers
1486

Species Strain * Location Host

SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
A. cucurbitae CBS 483.81 New Zealand Cucumis sativus KC584616 KC584358 FJ266483 AY562418 KC584483 KC584743 - - -
A. cumini CBS 121329 India Cuminum cyminum KC584523 KC584267 KC584191 KC584110 KC584391 KC584650 - - -
A. dennisii CBS 110533 New Zealand Jacobaea vulgaris KC584586 KC584328 KC584232 KC584157 KC584453 KC584712 - - -
T
A. dennisii CBS 476.90 Isle of Man Jacobaea vulgaris KC584587 KC584329 JN383488 JN383469 KC584454 KC584713 - - -
A. dianthicola CBS 116491 New Zealand Dianthus x allwoodii KC584526 KC584270 KC584194 KC584113 KC584394 KC584653 - - -
A. didymospora CBS 766.79 Mediterranean seawater KC584588 KC584330 FJ357312 FJ357300 KC584455 KC584714 - - -
T
A. eichhorniae CBS 489.92 India Eichhornia crassipes KP125049 NG_042763 NR_111832 KP124276 KP124895 KP125204 KP123973 KP124130 KP124740
A. eichhorniae CBS 119778 Indonesia Eichhornia crassipes KP125050 KP124580 KP124426 KP124277 KP124896 KP125205 - KP124131 KP124741
T
A. elegans CBS 109159 Burkina Faso Solanum lycopersicum KC584527 KC584271 KC584195 KC584114 KC584395 KC584654 - - -
A. embellisia CBS 339.71 USA Allium sativum KC584582 KC584324 KC584230 KC584155 KC584449 KC584708 - - -
A. eryngii CBS 121339  - Eryngium sp. KC584529 KC584273 JQ693661 AY562416 KC584397 KC584656 - - -
A. euphorbiicola CBS 198.86 USA Euphorbia pulcherrima - - KJ718172 KJ718017 KJ718345 KJ718520 KJ718686 - -
A. euphorbiicola CBS 109410  - Beta vulgaris MK249665 MK249662 KJ718173 - - KJ718521 - - -
T
A. eureka CBS 193.86 Australia Medicago rugosa KC584589 KC584331 JN383490 JN383471 KC584456 KC584715 - - -
A. gaisen CBS 632.93 Japan Pyrus pyrifolia KC584531 KC584275 KC584197 KC584116 KC584399 KC584658 KP123974 AY295033 KP124742
T
A. gaisen CBS 118488 Japan Pyrus pyrifolia KP125051 KP124581 KP124427 KP124278 KP124897 KP125206 KP123975 KP124132 KP124743
A. gaisen CPC 25268 Portugal unknown KP125052 KP124582 KP124428 KP124279 KP124898 KP125207 KP123976 KP124133 KP124744
A. gaisen MAFF 731001 Japan Fragaria sp. - - LC164854 LC169125 LC169131 LC167148 LC276235 LC276246 -
A. gaisen NCYUCC 19-0351 Taiwan Ficus microcarpa MT416127 MT416136 MT416122 MT945064 MT945070 MT420727 MT945062 MT945066 MT945068
A. gaisen NCYUCC 19-0353 Taiwan Kandelia candel MT416128 MT416137 MT416123 MT945065 MT945071 MT420728 MT945063 MT945067 MT945069
T
A. geniostomatis CBS 118701 New Zealand Geniostoma sp. KC584532 KC584276 KC584198 KC584117 KC584400 KC584659 - - -
T
A. gossypina CBS 104.32 Zimbabwe Gossypium sp. KP125054 KP124584 KP124430 JQ646312 KP124900 KP125209 JQ646395 KP124135 KP124746
T
A. gossypina CBS 102597 USA Minneola tangelo KP125056 KP124586 KP124432 KP124281 KP124902 KP125211 KP123978 KP124137 KP124748
T
ex-type strain, strain in this study are in bold.
Chiang Mai J. Sci. 2021; 48(6)
Table 1. (Continued).
GenBank accession numbers
Species Strain * Location Host
SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
T
A. gypsophilae CBS 107.41 Netherlands Gypsophila elegans KC584533 KC584277 KC584199 KC584118 KC584401 KC584660 JQ646387 - -
A. helianthiinficiens CBS 117370 England Helianthus annuus KC584534 KC584278 KC584200 KC584119 KC584402 KC584661 - - -
T
A. helianthiinficiens CBS 208.86 USA Helianthus annuus KC584535 KC584279 JX101649 KC584120 KC584403 EU130548 - - -
A. heterospora CBS 123376 China Solanum lycopersicum KC584621 KC584363 KC584248 KC584176 KC584488 KC584748 - - -
T
A. hyacinthi CBS 416.71 Netherlands Hyacinthus orientalis KC584590 KC584332 KC584233 KC584158 KC584457 KC584716 - - -
Chiang Mai J. Sci. 2021; 48(6)

T
A. indefessa CBS 536.83 USA soil KC584591 KC584333 KC584234 KC584159 KC584458 KC584717 - - -
T
A. infectoria CBS 210.86 England Triticum aestivum KC584536 KC584280 DQ323697 AY278793 KC584404 KC584662 - - -
T
A. iridiaustralis CBS 118486 Australia Iris sp. KP125059 KP124589 KP124435 KP124284 KP124905 KP125214 KP123981 KP124140 KP124751
A. iridiaustralis CBS 118487 Australia Iris sp. KP125060 KP124590 KP124436 KP124285 KP124906 KP125215 KP123982 KP124141 KP124752
T
A. iridicola MAFF 246890 Japan Iris japonica - - LC269974 LC270142 LC275240 LC275060 LC276238 LC276253 -
A. iridicola MUCC 2149 Japan Iris japonica - - LC269975 LC270143 LC275241 LC275061 LC276239 LC276254 -
T
A. jacinthicola CBS 133751 Mali Eichhornia crassipes KP125062 KP124592 KP124438 KP124287 KP124908 KP125217 KP123984 KP124143 KP124754
A. jacinthicola CPC 25267 Unknown Cucumis melo var. inodorus KP125063 KP124593 KP124439 KP124288 KP124909 KP125218 KP123985 KP124144 KP124755
Brassica rapa subsp.
A. japonica CBS 118390 USA KC584537 KC584281 KC584201 KC584121 KC584405 KC584663 - - -
Chinensis
A. kulundii M313 Russia soil KJ443087 KJ443132 KJ443262 KJ649618 KJ443176 - - - -
T
A. leucanthemi CBS 421.65 Netherlands Leucanthemum maximum KC584605 KC584347 KC584240 KC584164 KC584472 KC584732 - - -
A. leucanthemi CBS 422.65 USA  - KC584606 KC584348 KC584241 KC584165 KC584473 KC584733 - - -
A. limaciformis CBS 481.81 England  - KC584539 KC584283 KC584203 KC584123 KC584407 KC584665 JQ646368 - -
A. longipes CBS 113.35 Unknown Nicotiana tabacum KP125064 KP124594 KP124440 KP124289 KP124910 KP125219 KP123986 KP124145 KP124756
A. longipes CBS 121333 USA Nicotiana tabacum KP125068 KP124598 KP124444 KP124293 KP124914 KP125223 KP123990 KP124150 KP124761
T
A. macrospora CBS 117228 USA Gossypium barbadense KC584542 KC584286 KC584204 KC584124 KC584410 KC584668 KJ718702 - -
T
A. nepalensis CBS 118700 Nepal Brassica sp. KC584546 KC584290 KC584207 KC584126 KC584414 KC584672 - - -
A. nobilis CBS 116490 New Zealand Dianthus caryophyllus KC584547 KC584291 KC584208 KC584127 KC584415 KC584673 JQ646385 - -
T
ex-type strain, strain in this study are in bold.
1487
Table 1. (Continued).
GenBank accession numbers
1488

Species Strain * Location Host

SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
T
A. obclavata CBS 124120 USA air KC584575 FJ839651 KC584225 KC584149 KC584443 KC584701 - - -
T
A. omanensis SQUCC 13580 Oman dead woods MK878559 MK878556 MK878562 MK880899 MK880893 MK880896 - - -
A. omanensis SQUCC 15561 Oman dead woods MK878561 MK878558 MK878564 MK880901 MK880895 MK880898 - - -
T
A. oudemansii CBS 114.07  -  - KC584619 KC584361 FJ266488 KC584175 KC584486 KC584746 FJ266514 - -
A. panax CBS 482.81 USA Aralia racemosa KC584549 KC584293 KC584209 KC584128 KC584417 KC584675 - - -
T
A. papavericola CBS 116606 USA Papaver somniferum KC584579 KC584321 MH862999 FJ357298 KC584446 KC584705 - - -
A. penicillata CBS 116608 Austria Papaver rhoeas KC584572 KC584316 FJ357311 FJ357299 KC584440 KC584698 - - -
T
A. penicillata CBS 116607 Austria Papaver rhoeas KC584580 KC584322 KC584229 KC584153 KC584447 KC584706 - - -
A. perpunctulata CBS 115267 USA Alternanthera philoxeroides NG_062895 KC584294 NR_151838 KC584129 KC584418 KC584676 JQ905111 - -
A. petroselini CBS 112.41  - Petroselinum crispum KC584551 KC584295 KC584211 KC584130 KC584419 KC584677 - - -
A. petuchovskii M304 Russia alkaline soil KJ443079 KJ443124 KJ443254 KJ649616 KJ443170 - - - -
T
A. photistica CBS 212.86 England Digitalis purpurea KC584552 KC584296 KC584212 KC584131 KC584420 KC584678 - - -
T
A. phragmospora CBS 274.70 Netherlands soil KC584595 KC584337 MH859605 - KC584462 KC584721 - - -
A. porri CBS 116698 USA Allium cepa KC584553 KC584297 DQ323700 KC584132 KC584421 KC584679 KJ718726 - -
T
A. porri CBS 116699 - - KJ718218 KJ718053 KJ718391 KJ718564 KJ718727 - -
T
A. proteae CBS 475.90 Australia Protea KC584597 KC584339 AY278842 KC584161 KC584464 KC584723 FJ266505 - -
T
A. pseudorostrata CBS 119411 USA Euphorbia pulcherrima KC584554 KC584298 JN383483 AY562406 KC584422 KC584680 AY563295 - -
T
A. radicina CBS 245.67 USA Daucus carota KC584555 KC584299 KC584213 KC584133 KC584423 KC584681 FN689405 - -
A. scirpicola CBS 481.90 England Scirpus sp. KC584602 KC584344 KC584237 KC584163 KC584469 KC584728 - - -
T
A. septorioides CBS 106.41 Netherlands Reseda odorata KC584559 KC584303 KC584216 KC584136 KC584427 KC584685 - - -
A. septospora CBS 109.38 Italy wood KC584620 KC584362 FJ266489 FJ266500 KC584487 KC584747 FJ266515 - -
A. shukurtuzii M307 Russia alkaline soil KJ443082 KJ443127 KJ443257 KJ649620 KJ443172 - - - -
T
A. simsimi CBS 115265 Argentina Sesamum indicum KC584560 KC584304 JF780937 KC584137 KC584428 KC584686 JQ905110 - -
T
ex-type strain, strain in this study are in bold.
Chiang Mai J. Sci. 2021; 48(6)
Table 1. (Continued).
GenBank accession numbers
Species Strain * Location Host
SSU LSU ITS gapdh rpb2 ef1α Alta1 endoPG OPA10-2
T
A. slovaca CBS 567.66 Slovakia Homo sapiens KC584576 KC584319 KC584226 KC584150 KC584444 KC584702 - - -
A. smyrnii CBS 109380 England Smyrnium olusatrum KC584561 KC584305 AF229456 KC584138 KC584429 KC584687 - - -
A. solani CBS 116651 USA Solanum tuberosum KC584562 KC584306 KC584217 KC584139 KC584430 KC584688 - - -
T
A. soliaridae CBS 118387 USA soil KC584563 KC584307 KC584218 KC584140 KC584431 KC584689 - - -
T
A. solidaccana CBS 118698 Bangladesh soil KC584564 KC584308 KC584219 KC584141 KC584432 KC584690 - - -
Chiang Mai J. Sci. 2021; 48(6)

A. sonchi CBS 119675 Canada Sonchus asper KC584565 KC584309 KC584220 KC584142 KC584433 KC584691 - - -
A. tagetica CBS 479.81 England Tagetes erecta KC584566 KC584310 KC584221 KC584143 KC584434 KC584692 KJ718761 - -
T
A. tellustris CBS 538.83 USA soil KC584598 KC584340 MH861643 AY562419 KC584465 KC584724 AY563325 - -
T
A. thalictrigena CBS 121712 Germany Thalictrum sp. KC584568 KC584312 EU040211 KC584144 KC584436 KC584694 - - -
A. tomato CBS 103.30 Unknown Solanum lycopersicum KP125069 KP124599 KP124445 KP124294 KP124915 KP125224 KP123991 KP124151 KP124762
A. tomato CBS 114.35 Unknown Solanum lycopersicum KP125070 KP124600 KP124446 KP124295 KP124916 KP125225 KP123992 KP124152 KP124763
T
A. triglochinicola CBS 119676 Australia Cycnogeton procerum KC584569 KC584313 KC584222 KC584145 KC584437 KC584695 - - -
A. vaccariae CBS 116533 USA Vaccaria hispanica KC584570 KC584314 KC584223 KC584146 KC584438 KC584696 JQ646386 - -
Alternaria sp. CBS 198.67 USA soil KC584610 KC584352 AF229487 KC584169 KC584477 KC584737 - - -
Alternaria sp. CBS 115.44  - Reseda odorata KC584556 KC584300 KC584214 KC584134 KC584424 KC584682 - - -
T
Pleospora tarda CBS 714.68 Canada  - KC584603 KC584345 KC584238 - - KC584729 - - -
Stemphylium herbarum CBS 191.86 India  - GU238232 GU238160 KC584239 - KC584471 KC584731 - - -
T
ex-type strain, strain in this study are in bold.
1489
 220 leaves as small circular spots. Asexual morph: Hyphomycete. Conidiophores 50–100
221 simple or sparingly branched, straight to slightly curved, pale to medium brown, w
222 short, geniculate, sympodial proliferations, gradually enlarging near the apex
223 conidiogenous cell, commonly with a terminal cluster of 1–3 conidia. Conidiogeno
1490 224 polytretic, clavate, smooth, pale brown, Chiang
withMai mediumJ. Sci.brown
2021;cicatrized
48(6) conidiogen
225 concatenated conidia, 1–3+ conidia. Conidia (15–)20–30(–43) × (9–)11–13(–14) μm
226 μm, n=30), conspicuously obovoid to spheroid, beaked, pale to dark brown, with 2–
227 and 1–2 longitudinal or oblique septa, constricted at the septa, usually solitary, st
228 curved, with a long ellipsoid to obclavate, simple, pale brown, up to 10–12 μm long
3476–3490, 3491–3901, 3902–4069, 229 4070–4293,
tapering to beaked Index
apex.Fungorum number: IF252306.
and 4294–4352 corresponding to 230SSU, LSU,Associated
ITS1+ with Facesoffungi
leaf spot ofnumber:
KandeliaFoF08022.
candel. Symptoms small irregular to subci
ITS2, 5.8S, gapdh-exon, rpb2, 231 brown,
ef1α-exon, slightly sunken≡spots
Alta1- appeargaisen
Alternaria on adaxial
Nagano, surface leaves
J. Hort. of K.
Soc. candel, which later
Japan
232 on the
exon, gapdh-intron, ef1α-intron, and Alta1-intron,surface of the leaves. Small
32(3): 16-19 (1920) auburn spots appeared initially and then gradually e
233 to tawny circular ring spots with a dark mahogany border and jagged edge. They
respectively. 3063 constant characters,
234 246 variable
circulars, which occurred = Alternaria
on a singlekikuchiana S. Tanaka,
affected leaf. In severe Memoirs of
cases, lesions spread eve
characters are parsimony-uninformative,
235 Notand 1043 fungal
produce thestructure
Coll. Agric. Kyoto
on leaf Imper.
spot symptom.Univers., Phytopathol.
236
characters are parsimony-informative. The trees Ser. 28: 27 (1933)
237 Culture characteristics: Colonies on PDA develop well, attaining a diameter of
generated under different optimality criteria based = Macrosporium nashi Miura, Flora of Manchuria
238 a daily fluorescent light/dark cycle of 8/16 h at 25 °C in 7 d, circular, with evident co
on MP, ML and BI analysis were239 essentially similar and East Mongolia,
a whitish aerial-mycelial margin, III Cryptogams,
cottony, greenish to light Fungi (Industr.
grey, secreting an orangish
in topology and not significantly240
different(data
medium not Contr.
(Figure 3P). S. Manch.
Mycelium 3–4 Rly
μm 27):
diam,513 (1928)
hyaline to pale-brown, smooth, septat
arise singly
241generated
shown). The descriptive statistics fromfrom vegetativeFungus hyphae, branches,
associated with1–2leafconidiogenous
blight of Ficus loci, each conidioge
242 concatenated conidia, 1–5+ conidia, the obvious branching is initiated within the c
MP analysis, TL = 5208, CI = 243 0.382, RI = 0.709, microcarpa. Symptoms appeared as circular spots
secondary conidiophores, intercalary conidial pseudorostra, develop with 2–3 co
RC = 0.271, and HI = 0.618.244 Conidia in cultureon
The best scoring adaxial and abaxial
(15–)22–28(–44) surface of the
× (6–)9–12(–18) μm leaves,
(x̅ = 25 × light
11.5 μm, n = 30) a
likelihood tree selected with a 245 agar for
final value surface
the or directly
brown withfromahyphae,
pale lightformed
center,acropetal
with fungus chains, conspicuously obo
spores
combined dataset = -31969.718266.246 tapering to beaked apex, pale
forming to dark brown,
concentric brownwith 2–4astransverse
rings they enlarge, septa and 1–2 longi
247 septa, with outer wall usually smooth, but sometimes are detectably rough
The phylogenetic tree in Figure
248 2 included
occasionally41 definitely
with lesions tending
tuberculate to appear
(Figure 3Q–T).first on the margin
strains, with Alternaria perpunctulata
249 (CBS 115267)
Colonies on PCA similar to those observedspots.
of leaves as small circular on PDA Asexual
lesion,morph:
attaining a diameter of
250 a daily
and A. alternantherae (CBS 124392) as outgroup fluorescent light/dark cycle of 8/16
Hyphomycete. Conidiophores 50–100h at 25 °C in 7 d,µm circular,
long,with evident co
251 a whitish
taxa. The total length of the Figure 2 dataset aerial-mycelial
septate, simple or sparingly branched, straight2–4
margin, greenish to light grey. Mycelium to μm diam., pal
252 septate. Conidiophores arise singly from vegetative hyphae, branches, 1–3 co
was 5293 characters including253 alignment gaps,1–3 conidial
(therefore slightlychains),
curved,eachpaleconidiogenous
to medium brown, branchwith withseries
concatenated conidi
1–1020, 1021–1585, 1586–1912, 254 1913–2070, of 6–12 short,
Conidia (10–)22–25(–30) geniculate, sympodial
× (6–)10–12(–13) μm (x̅ = 22proliferations,
μm × 10 μm, n=30), formed
2071–2478, 2479–3338, 3339–3353, ellipsoid when transverse-septate
255 3354–3764, gradually enlarging beginning
near tothe form,
apex obovoid
into atoclavate
spheroid at stage of d
256 2–3 transverse septa, with 1–3 longitudinal septate (almost 1-septate), smooth, second
3765–4208, 4209–4842, 4843–5008, 5009–5233, conidiogenous cell, commonly with a terminal
257 with short beaked apex (Figure 4A–F).
and 5234–5293 corresponding to 258SSU, LSU,Colonies
ITS1+ on cluster
V-8 agarofdevelop
1–3 conidia. Conidiogenous
well, attaining cells mono-
a diameter of 7.5–8 cm under a
ITS2, 5.8S, gapdh-exon, rpb2, 259 ef1α-exon, Alta1-
light/dark to8/16
cycle of polytretic,
h at 25 °Cclavate,
in 7 d,smooth,
circular,palewith brown, with
evident concentric rings and
260 mycelial
exon, endoPG, OPA10-2, gapdh-intron, ef1α-intron margin, cottony, greenish light grey to
medium brown cicatrized conidiogenous loci, white at above, secreting an gree
261 pigment into the medium. Mycelium 3–5 μm diam., hyaline to pale-brown,
, and Alta1-intron, respectively.262
TheConidiophores
combined arise produce concatenated conidia, 1–3+ conidia.
singly from vegetative hyphae, branches, 1–2 conidiogenous l
dataset contained 4734 constant, 263114 parsimony- Conidia (15–)20–30(–43)
chains, each conidiogenous × (9–)11–13(–14)
branch with concatenated conidia, 1–5+. μm Conidia (15–)
264 )10–12(–13) μm (( x̅ ==2424μm
uninformative and 445 parsimony-informative μm × 11
× μm,
11.5n=30),
μm, formed
n=30), acropetal
conspicuously chains, ellipsoid when
characters. The combined dataset 265 was beginning
analyzedto form, obovoidtotospheroid,
obovoid spheroid beaked,
at stage of distal
pale cell brown,
to dark produces 2–5 transvers
using MP, ML and BI. The trees generated under with 2–6 transverse septa and 1–2 longitudinal
different optimality criteria were essentially similar or oblique septa, constricted at the septa, usually
in topology and not significantly different (data solitary, straight or slightly curved, with a long
not shown). The descriptive statistics generated ellipsoid to obclavate, simple, pale brown, up to
from MP analysis based on the combined dataset 10–12 μm long and 2–3 μm wide, tapering to
were TL = 791, CI = 0.774, RI = 0.890, RC = beaked apex.
0.688, HI = 0.226. The best scoring likelihood Associated with leaf spot of Kandelia candel.
tree selected with a final value for the combined Symptoms small irregular to subcircular shape, pale
dataset = -12352.288546. brown, slightly sunken spots appear on adaxial
surface leaves of K. candel, which later expand
3.2 Taxonomy outwards on the surface of the leaves. Small
Alternaria gaisen Nagano ex Hara, Sakumotsu auburn spots appeared initially and then gradually
byorigaku, Edn 4: 263 (1928) (Figure 3 and 4) enlarged, changing to tawny circular ring spots
MycoBank no: MB 252306. with a dark mahogany border and jagged edge.
Chiang Mai J. Sci. 2021; 48(6) 1491

Figure 3. Alternaria gaisen (NCYUCC 19-0351). (a, b) Habitat; (c) Leaf blight on Ficus microcarpa; (d-g)
Conidiomata on host surface; (h, i) Conidiomata; (j-o) Conidia; (p) Culture characters on PDA (left-
above, right-reverse); (q-t) Mycelium with conidia from cultures on PDA. Bars: d 2000 mm; e 500 µm;
f 300 µm; g 100 µm; h 50 µm; i, j, q, s, t 20 µm; k-o, r 10 µm.
1492 Chiang Mai J. Sci. 2021; 48(6)

Figure 4. Alternaria gaisen (NCYUCC 19-0351). (a, b) Culture characters on PCA with 14 days (A-above,
B-reverse); (c) Mycelium with conidia from cultures on PCA; (d-f) Conidia, conidiophores, and
sporulation patterns from cultures on PCA; (g, h) Culture characters on V-8 agar with 14 days (G-above,
H-reverse); (i, j) Mycelium with conidia from cultures on V-8 agar; (k, l) Conidia, conidiophores, and
sporulation patterns from culture on V-8 agar. Bars: C, I, J 200 µm; D–F, K, L 20 µm.
 238 a daily fluorescent light/dark cycle of 8/16 h at 25 °C in 7 d, ci
own, slightly sunken spots appear on adaxial surface leaves of K. candel,239 which alater expand
whitish outwards margin, cottony, greenish to light gre
aerial-mycelial
n the surface of the leaves. Small auburn spots appeared initially and then240 gradually enlarged,
medium (Figure changing
3P). Mycelium 3–4 μm diam, hyaline to pale
tawny circular ring spots with a dark mahogany border and jagged 241 edge. They were usually
arise singly from vegetative >3 hyphae, branches, 1–2 conidiogen
Chiang Mai
rculars, which occurred on a J.single
Sci. 2021;
affected 48(6)
leaf. In severe cases, lesions242 spreadconcatenated
evenly on theconidia, leaves. 1–5+ conidia, the 1493 obvious branching i
ot produce fungal structure on leaf spot symptom. 243 secondary conidiophores, intercalary conidial pseudorostra,
= Alternaria kikuchiana S. Tanaka, Memoirs of the Coll. Agric. Kyoto 244Imper. Univers.,
Conidia in culture (15–)22–28(–44) × (6–)9–12(–18) μm (x̅ =
Culture characteristics:
hytopathol. Ser. 28: 27 (1933) Colonies on PDA develop well, attaining a diameter 245 agar7.3–7.5
of surfacecm or under
directly from hyphae, formed acropetal chai
daily fluorescent light/dark
= Macrosporium cycle Flora
nashi Miura, of 8/16 ofhManchuria
at 25 °C inand 7 d,East
circular,
Mongolia,with246
evident concentric
III Cryptogams,
tapering to beakedand
rings
Fungi apex,
whitish Contr. They
aerial-mycelial were
S. Manch.margin, usually >3
cottony, circulars, which occurred
greenish to light grey, secreting247 secondary
an orangish conidiophore withpale to dark
short brown,
beaked apexwith 2–4 transve
ndustr. Rly 27): 513 (1928) septa,pigmentwith outer into the wall usually smooth, but sometimes a
edium (Figure 3P). on aMycelium
single affected
3–4 μmleaf. diam, Inhyaline
severe to cases, lesions smooth,
pale-brown, (Figure
248 septate.
4A–F).
occasionally Conidiophores
definitely tuberculate (Figure 3Q–T).
iseFungus
singly from vegetative
associated
spread with hyphae,
leaf on
evenly branches,
blight
the of Ficus
leaves. 1–2 conidiogenous
microcarpa.
Not produceSymptoms loci, each
fungal 249 conidiogenous
appeared as
Colonies circular
Colonies branch
on spots
V-8
on PCA with
onsimilar
agar develop well, attaining
to those observed on PDA lesion,
ncatenated
axial conidia,surface
and abaxial 1–5+ conidia,
of the the obvious
leaves, light branching
brown with isa initiated
pale lightwithin
center,the chain
with when
fungus a few
spores
structure on leaf spot symptom. 250a diameter
a dailyof 7.5–8 cmlight/dark
fluorescent under a daily
cyclefluorescent
of 8/16 h at 25 °C in 7 d, ci
condary conidiophores,
rming concentric brown ringsintercalary
as theyconidial
enlarge,pseudorostra,
with lesions tending developtowith appear
251 2–3afirst
conidiogenous
on the
whitish margin loci.
aerial-mycelial of margin, greenish to light grey. Myce
aves asinsmall
onidia culture
circular Culture
spots. characteristics:
(15–)22–28(–44) Asexual morph:Colonies
× (6–)9–12(–18) on
μmPDA
Hyphomycete. (x̅ = develop
25 × 11.5 μm,
Conidiophores light/dark
n = 30)
50–100 cycle
arising
µm of septate,
8/16theh at 25
beneath
long, °C in 7 d, circular,
252 septate. Conidiophores arise singly from vegetative hyph
ar surface
mple or sparingly well,
or directly attaining
branched, a diameter
from hyphae,
straight formed
to of acropetal
slightly 7.3–7.5
curved,cm paleunder
chains, a 253
to conspicuously
medium with
brown, evident
obovoid
with series
(therefore toconcentric
1–3spheroid,
ofconidial rings and
6–12 chains), eacha conidiogenous
whitish branch w
pering to beaked
ort, geniculate, apex,
daily pale to proliferations,
sympodial
fluorescent darklight/dark
brown, with 2–4 transverse
gradually
cycle 8/16 septa
of enlarging h at near and
254 1–2
the longitudinal
apex
aerial-mycelial into or
a oblique
clavate
margin, cottony,
Conidia (10–)22–25(–30) × (6–)10–12(–13) greenish light
μm (x̅ = 22 μm ×
pta, with outer
nidiogenous cell, wall usually smooth, but cluster
sometimes areconidia.
detectably rough (pusticulate) and
casionally
25 commonly
definitely
in 7 d,with
°Ctuberculate a terminal
circular,
(Figurewith
with
3Q–T).
evident of 1–3
concentric Conidiogenous
255grey to ellipsoid
white cells
when mono-
at above, to
transverse-septate
secreting anbeginning
greenish to toform, obovoid t
olytretic, clavate, smooth, pale brown, medium brown cicatrized256 conidiogenous
2–3 loci,
transverse produce
septa, with
Colonies on
ncatenated ringssimilar
PCA
conidia, and aconidia.
1–3+ whitish aerial-mycelial
to thoseConidia
observed on PDA margin,
(15–)20–30(–43)lesion,cottony,
attaining a257
× (9–)11–13(–14) light grey
diameter μmof (x ̅ pigment
7.8–8.2 into
cm×under
= 24beaked
μm 11.5 the medium. Mycelium (almost 1-s
1–3 longitudinal septate
with short apex (Figure 4A–F).
daily
m, fluorescent
n=30), greenish
light/dark
conspicuously tocycle
obovoidlight to grey,
of 8/16 h secreting
spheroid, at 25 °C in an
beaked, d,orangish
7pale circular,
to dark with brown, 3–5
evident
258 withμm diam.,
concentric
2–6 transverse
Colonies hyaline
rings and
on septa
V-8 toagar
pale-brown, smooth,
develop well, attaining a diameter
whitish aerial-mycelial
d 1–2 longitudinal pigment margin,
or oblique
into the greenish
septa,
medium to light
constricted
(Figuregrey. Mycelium
at 3P).
the septa,
Mycelium 2–4 μm
usually diam.,light/dark
solitary,
259septate. pale-brown,
straight
Conidiophores or smooth,
cycleslightly
ofarise
8/16singly from
h at 25 vegetative
°C in 7 d, circular, with evide
ptate. Conidiophores
rved, with a long ellipsoid ariseto singly
obclavate, from vegetative
simple, hyphae,
pale brown, up to branches,
10–12 μm 1–3long conidiogenous
and 2–3
mycelial μm wide,
margin, loci
herefore
pering to1–3
3–4
conidial
beaked
μm diam, hyaline to pale-brown, smooth, 260
apex.chains), each conidiogenous branch with concatenated
hyphae,
conidia, branches,
1–10+ 1–2cottony,
conidia.
greenish light
conidiogenous grey to white at ab
loci with
261 pigment into the medium. Mycelium 3–5 μm diam., hy
septate.
(10–)22–25(–30) Conidiophores
× (6–)10–12(–13) arise singly
μm =from
(x̅ Symptoms
22 μm vegetative
×small
10 μm, n=30),conidial chains, each conidiogenous
palesingly from branch with
onidia
Associated with leaf spot of Kandelia candel. 262 toformed
irregular acropetal
subcircular
Conidiophores shape,chains,
arise vegetative hyphae, branche
lipsoid when transverse-septate
own, slightly sunken hyphae, spotsbranches, beginning
appear on adaxial to form,
1–2 conidiogenous obovoid to
surface leaves of K.loci, spheroid at
candel,263 stage of
concatenated
which distal
later expand
chains, cell
conidia,
each produces
1–5+. Conidiabranch
outwards
conidiogenous (15–)22–25(–35)
with concatenated conidia
–3the
n transverse
surface of septa,
the
each with
leaves. 1–3 longitudinal
Small
conidiogenous auburnbranch septate
spots (almost
appeared
with 1-septate),
initially
concatenated and then smooth, secondary
gradually
264 enlarged,
× (6–)10–12(–13)
)10–12(–13) conidiophore
changing
μm (( x̅ ==24
μm 24μm
μm××11 11μm,
μm,n=30),
n=30),formed acropeta
ith shortcircular
tawny beaked apex (Figure
ring spots 4A–F).
with a dark mahogany border and jagged 265 edge. They were to
beginning usually
form, >3
obovoid to spheroid at stage of distal cell
Colonies
rculars, which conidia,
on occurred
V-8 agaron 1–5+
develop conidia,
a single well, theleaf.
obvious
attaining
affected branching
a severe
In diameter cases,of 7.5–8is cmspread
lesions formed aacropetal
underevenly daily thechains,
leaves.ellipsoid when transvers
onfluorescent
ght/dark
ot produce cycle initiated
of
fungal 8/16 within
h at
structure 25on°C the
leaf 7chain
inspot when with
d,symptom.
circular, a fewevident
secondary concentricseptaterings and beginning
a whitishtoaerial- form, obovoid to spheroid
ycelial margin,conidiophores,
cottony, greenish light grey to white
intercalary conidial pseudorostra, at above, secreting at stage of distal cellgrey
an greenish to light produces 2–5 transverse
gment intocharacteristics:
the medium. Colonies Mycelium on 3–5
PDA μm
develop diam.,
well, hyaline
attaining to apale-brown,
diameter smooth,cm septate.
Culture develop with 2–3 conidiogenous loci. Conidia in septateof(mostly7.3–7.5 under 1–2 longitudinal septate
3-septate),
onidiophores
daily fluorescent arise singly from
light/dark cyclevegetative
of 8/16 h at hyphae,
25 °C in branches,
7 d, circular,1–2 conidiogenous loci withrings
with evident concentric conidial
and
ains, each
whitish culture margin,
conidiogenous
aerial-mycelial (15–)22–28(–44)
branch cottony, × (6–)9–12(–18)
with concatenated
greenish conidia,
to light grey,1–5+. μm Conidia
secreting (almost
an 1-septate),
(15–)22–25(–35)
orangish pigment into secondary
× (6–
the conidiophore, with
0–12(–13)
edium (Figure ( x̅ ==Mycelium
μm (3P). 2425μm× ×11.511
3–4 μm,
μm, μm ndiam,
n=30),= 30) arising
formed
hyaline tobeneath
acropetal the smooth,
chains,
pale-brown, smooth,
ellipsoid whenshort
septate. transvers beakedseptate
Conidiophores apex (Figure 4G–L).
ginning
ise singlytofrom
form, obovoid
vegetative
agar surfaceto spheroid
hyphae, at stage
branches,
or directly 1–2
from ofconidiogenous
distal cellformed
hyphae, produces
loci, each2–5conidiogenous
transverse
Materialseptate branch
examined: (mostly
with
TAIWAN, Tainan, Bei Men
ncatenated conidia, 1–5+ conidia, the obvious branching is initiated within the chain when a few
acropetal chains, conspicuously obovoid to spheroid, Mangrove Reserve, on leaf of Ficus microcarpa,
condary conidiophores, intercalary conidial pseudorostra, develop with 2–3 conidiogenous loci.
onidia in culturetapering to beaked×apex,
(15–)22–28(–44) pale to dark
(6–)9–12(–18) μmbrown,
(x̅ = 25 × with
11.5 μm,15 n =Jul30)2018,
arising Chada
beneath Norphanphoun
the BM2, dried
2–4 transverse
ar surface or directly from hyphae, septa and 1–2
formed longitudinal
acropetal or
chains, conspicuously cultureobovoid
(NCYUCC 19-0351); Taichung, on leaves of
to spheroid,
pering to beaked apex, pale
oblique septa,to dark
with brown,
outer wall withusually
2–4 transverse
smooth,septa but and Kandelia
1–2 longitudinal
candel, 15orJuloblique 2018, Chada Norphanphoun
pta, with outer wall usually smooth, but sometimes are detectably rough (pusticulate) and
sometimes are detectably
casionally definitely tuberculate (Figure 3Q–T).
rough (pusticulate) and TC1-B (NCYUCC 19-0353,), dried culture NCYU
Colonies on PCA occasionally
similar todefinitely
those observed tuberculate
on PDA (Figure
lesion,3Q–T). herbarium.
attaining a diameter of 7.8–8.2 cm under
daily fluorescent light/dark Colonies cycleonofPCA 8/16similar
h at 25 to °C those
in 7 d,observed
circular, with evident Hosts and distributions:
concentric rings and Chimonanthus praecox
whitish aerial-mycelial margin, greenish to light
on PDA lesion, attaining a diameter of 7.8–8.2 cm grey. Mycelium 2–4 μm diam., pale-brown,
(CHINA [52]), Fragaria smooth, × ananassa (JAPAN [32],
ptate. Conidiophores arise singly from vegetative hyphae, branches, 1–3 conidiogenous loci
under a daily fluorescent light/dark cycle of 8/16
herefore 1–3 conidial chains), each conidiogenous branch with concatenated conidia, 1–10+ conidia. h [33]), Oryza sativa (PAKISTAN, [1]), Pyrus aromatica
onidia (10–)22–25(–30)at 25 °C ×in(6–)10–12(–13)
7 d, circular, with μm (x evident
̅ = 22 μm concentric
× 10 μm, n=30), (JAPAN
formed [55]),
acropetal P. chains,
bretschneideri (CHINA [54]),
lipsoid when transverse-septate
rings and a whitish beginning to form, margin,
aerial-mycelial obovoid greenish
to spheroid at (CHINA stage of distal [51]), cellP.produces
communis (UNITED STATES
–3 transverse septa, to lightwithgrey.
1–3 longitudinal
Mycelium 2–4septate μm diam., (almost 1-septate), smooth,
pale-brown, [54]),secondary
P. pyrifolia conidiophore
(JAPAN [3], [15], [30], [47], [59],
ith short beaked apex (Figure 4A–F).
smooth, septate. Conidiophores arise singly from
Colonies on V-8 agar develop well, attaining a diameter of 7.5–8 cm under a daily fluorescent [60]), (KOREA [3], [62]), (TAIWAN [3]), P. pyrifolia
ght/dark cycle of vegetative
8/16 h athyphae,
25 °C in branches,
7 d, circular,1–3 conidiogenous loci
with evident concentric var.
ringsculta
and(JAPAN
a whitish [32], [33]), P. serotina (FRANCE
aerial-
ycelial margin,(therefore
cottony, greenish
1–3 conidial lightchains),
grey toeach white at above, secreting[6]),
conidiogenous an (JAPAN
greenish to [4],light
[55]), grey(KOREA [11]), Pyrus sp.
gment into the medium. Mycelium 3–5 μm diam., hyaline to pale-brown, smooth, septate.
branch with concatenated conidia, 1–10+ conidia. (CHINA [63], [64]), (UNITED STATES [49]),
onidiophores arise singly from vegetative hyphae, branches, 1–2 conidiogenous loci with conidial
Conidia (10–)22–25(–30) × (6–)10–12(–13)
ains, each conidiogenous branch with concatenated conidia, 1–5+. Conidia μm (TAIWAN),
(15–)22–25(–35) P. ussuriensis
× (6–(CHINA [51]).
0–12(–13) μm (( x̅ ==24 22μmμm × 10
× 11 μm,μm, n=30), n=30),
formed formed
acropetalacropetal
chains, ellipsoid when Notes:transvers
Alternariaseptategaisen was first reported from
ginning to form, obovoid
chains, to spheroid
ellipsoid at stage of distal cell
when transverse-septate produces 2–5black
beginning transverse septate of
spot disease (mostly
Japanese pear (Pyrus pyrifolia)
to form, obovoid to spheroid at stage of distal by Nagano (1920). Later, it was reported with a
cell produces 2–3 transverse septa, with 1–3 wide host range as fungal invaders of fruits and
longitudinal septate (almost 1-septate), smooth, leaves on Chimonanthus praecox (Calycanthaceae),
1494 Chiang Mai J. Sci. 2021; 48(6)

Oryza sativa (Poaceae), and Pyrus spp. (Rosaceae).
This species was placed in the section Alternaria
based on molecular analysis [25]. Our isolates
were collected from Bei-Men Mangrove Reserve, a
mangrove restoration area in Tainan, and mangrove
area in Taichung, Taiwan. Phylogenetic analyses
demonstrated that the new isolates are a known
species, A. gaisen, with high bootstrap support
(Figures 1 and 2). The morphology of the new
isolates is similar to A. gaisen with Alternaria-
shape and short beaked conidia in chain was
produced with 8–10 conidia, and branched at
1–3 conidiogenous loci (Table 2). However, the
conidial size of the new isolates was smaller than
A. gaisen, which is listed in Table 2. Phylogenetic
analyses and morphological characters indicated
that our isolates are A. gaisen. The morphological
characters of species related in this study are
mentioned in Table 2 [33], [46].
4. DISCUSSION
The new host record of Alternaria gaisen was
isolated from leaf blight of Ficus microcarpa and
leaf spots of Kandelia candel in Taiwan mangroves.
Alternaria species have overlapping morphological
traits even within section and species level.
Sequence data is therefore essential to identify
these species and clarify species sections. Previous
studies used multi-gene sequence data to provide
a better resolution for Alternaria species viz. SSU,
LSU, ITS, gapdh, rpb2, ef1α, Alta1, endoPG, and
OPA10-2 gene regions and else [2], [25–27],

Table 2. Comparison of morphological characteristics of Alternaria species related to this study.

Conidia
Species Media References
Size (µm) Shape Septation
A. arborescens 12–30(–42) Ovoid to ellipsoid, pale brown to 1–4 transverse, PCA Simmons [44]
×7–11 brown, formed conidial chains with 0–2 longitudinal
branched septa
A. alternata 10–30 × 5–12 Ovoid to ellipsoid, pale-yellow to 4–7 transverse PCA Simmons [44]
golden brown, formed conidial chains septa, a few, or no
with short secondary conidiophore, longitudinal septa
branched
A. gaisen (10–)22–25(–30) Ellipsoid when transverse septate 2–4 transverse PCA This study
× (6–)10–12(– beginning to form, obovoid to septa, with 1–3
13) spheroid at stage of distal cell longitudinal
produces secondary conidiophores, septate (almost
pale to dark brown, formed conidial 1-septate)
chains of 1–10+ conidia and
branched (1–3 conidial chain)
A. gaisen 30–45(–55) × Obclavate, oblong-ovate or rarely, 5–8 transverse, PCA Simmons [44]
13–15(–18) smooth, or sometimes verrucose, 0–1(–2)
olive-brown, formed conidial chains longitudinal septa
of 5–8 conidia and branched, 1–3 in each transverse
conidiogenous loci
A. iridicola 28–311 × 7–38 Ovoid, ellipsoid to broadly obclavate, 2–16 transverse PCA Nishikawa and
(total) or sometimes beakless small oval, pale septa, 0–11 Nakashima [26]
21–127 × 7–38 brown to yellowish brown, formed longitudinal septa
(bodies) conidial chains of 3–4 conidia,
unbranched
Chiang Mai J. Sci. 2021; 48(6) 1495

[33], [60]. In this study the combined gene trees
were considered, while greatly facilitated species
level identification provided better resolution.
Morphological and phylogenetic evidence with
a MEGA BLAST search showing percent of
identities (I), and query cover (QC) (Table 3).
Based on the results of our phylogenetic analyses
the new isolates clustered within sect. Alternaria
in Figure 1, and also grouped with A. gaisen and
A. iridicola species group with a high moderate
suport in figure 2 (99% MP/100% ML/1.0 BS),
which is the phylogenetic tree that included only
strains of sect. Alternaria. Alternaria iridicola was
distinguished from our isolates based on morphology
of ovoid, ellipsoid to broadly obclavate shaped
with sometimes beakless small oval and large
conidia (28–311 × 7–38 µm), and multi-transverse
septa (2–16), and longitudinal septa (0–11), with
unbranched conidial chains. The morphology of
A. alternata, and A. arborescens were compared to
determine the species identification, which were
similar to our isolates with conidial size (10–30
× 5–12 µm, 12–30(–42) ×7–11 µm, respectively),
number of transverse and longitudinal conidia
septa (Table 2 and Figure 4). The comparison was
based on an identification manual of Alternaria by
Simmons [47]. However, our isolates can easily be
distinguished from A. alternata and A. arborescens
using molecular data (Figure 2). Thus, our isolates
are identified as A. gaisen as mentioned the evidence
in notes of species.
This study has located a species of Alternaria
that associated with leaf blight of F. microcarpa and
leaf spot of K. candel in Taiwan mangroves. This
indicates that fungi on mangroves may act as an
inoculum for adjacent agricultural and horticultural
crops. It would be interesting to establish if
other fungi on mangroves can also act in this
way. The new collections of A. gaisen described
in this paper was found from mangrove plants in
Taiwan and this is the first record of A. gaisen on
F. microcarpa and K. candel in Taiwan. Many fungal
species were found in Taiwan’s mangroves in recent
years especially marine mangrove fungi [21], [36].
Pathogenicity testing on various hosts is needed
to confirm the pathogenic affinity of the taxon.

Table 3. GenBank BLAST search of Alternaria gaisen, a new record species in this study.
GenBank BLAST search against type material
Genes Accession Identities (I), Query
Species identified Strain no.
no. cover (QC)
SSU A. oudemansii (sect. Ulocladium) CBS 114.07 NG_067644 100% (I), 100% (QC)
A. botrytis (sect. Ulocladium) CBS 197.67 NG_067640 100% (I), 100% (QC)
A. leucanthemi (sect. Teretispora) CBS 421.65 NG_067639 100% (I), 100% (QC)
A. papavericola (sect. Crivellia) CBS 116606 NG_067636 100% (I), 100% (QC)
A. iridiaustralis (sect. Alternaria) CBS 118486 NG_063035 100% (I), 100% (QC)
LSU A. alstroemeriae (sect. Alternaria) CBS 118809 MH874589 100%(I), 99% (QC)
A. aspera (sect. Pseudoulocladium) CBS 115269 MH874542 99.66%(I), 99% (QC)
A. alternata (= A. angustiovoidea, sect. Alternaria)  CBS 195.86 MH873628 99.89%(I), 99% (QC)
A. helianthiinficiens CBS 208.86 MH873632 99.66%(I), 99% (QC)
A. planifunda (sect. Embellisia) CBS 537.83 MH873356 99.55%(I), 99% (QC)
1496 Chiang Mai J. Sci. 2021; 48(6)

Table 3. (Continued).
GenBank BLAST search against type material
Genes Accession Identities (I), Query
Species identified Strain no.
no. cover (QC)
ITS A. perpunctulata (sect. Alternantherae) CBS 115267 NR_151838 100% (I), 100% (QC)
A. alternata (= A. angustiovoidea, sect. Alternaria)  CBS 195.86 MH861939 94.46% (I), 100% (QC)
A. alternata (= A. destruens, sect. Alternaria) ATCC 204363 NR_137143 94.46% (I), 100% (QC)
A. iridiaustralis (sect. Alternaria) CBS 118486 NR_136120 94.26% (I), 100% (QC)
A. alstroemeriae (sect. Alternaria) CBS 118809 NR_163686 94.26% (I), 100% (QC)
gapdh A. arborescens (sect. Alternaria) CBS 102605 AY278810 99.66% (I), 97% (QC)
A. alternata (= A. daucifolii, sect. Alternaria) CBS 118812 KC584112 99.65% (I), 97% (QC)
A. alstroemeriae (sect. Alternaria) CBS 118809 KP124154 99.48% (I), 97% (QC)
A. alternata (= A. rhadina, sect. Alternaria) CBS 595.93 JQ646316 99.65% (I), 97% (QC)
A. alternata (= A. citriarbusti, sect. Alternaria) SH-MIL-8s JQ646322 99.65% (I), 97% (QC)
rpb2 A. pseudoeichhorniae (sect. Alternaria) MFLUCC 18-1589 MH853717 98.5% (I), 87% (QC)
A. arborescens (sect. Alternaria) CBS 102605 KC584377 98.5% (I), 87% (QC)
A. alternata (= A. daucifolii, sect. Alternaria) CBS 118812 KC584393 98.15% (I), 87% (QC)
A. prunicola (sect. Alternaria) MFLUCC 18-1597 MH853722 98.03% (I), 87% (QC)
A. alstroemeriae (sect. Alternaria) CBS 118809 KP124765 98.8% (I), 75% (QC)
ef1α A. alternata (= A. daucifolii, sect. Alternaria) CBS 118812 KC584652 98.33% (I), 99% (QC)
A. iridiaustralis (sect. Alternaria) CBS 118486 KP125214 97.88% (I), 98% (QC)
A. betae-kenyensis (sect. Alternaria) CBS 118810 KP125197 97.49% (I), 99% (QC)
A. burnsii (sect. Alternaria) CBS 107.38 KP125198 97.49% (I), 99% (QC)
A. eichhorniae (sect. Alternaria) CBS 489.92 KP125204 97.07% (I), 99% (QC)
Alta1 A. alstroemeriae (sect. Alternaria) CBS 118809 MH084526 97.46% (I), 89% (QC)
A. acalyphicola (sect. Porri) CBS 541.94 JQ905109 97.25% (I), 89% (QC)
A. agripestis (sect. Porri) CBS 577.94 JQ905112 97.03% (I), 89% (QC)
A. aragakii (sect. Porri) CBS 594.93 JQ905107 97.03% (I), 89% (QC)
A. alternata (= A. rhadina, sect. Alternaria) CBS 595.93 JQ646399 97.03% (I), 89% (QC)
endoPG A. arborescens (sect. Alternaria) EGS 39.128 AY295028 96.95% (I), 100% (QC)
A. alstroemeriae (sect. Alternaria) CBS 118809 KP123994 97.32% (I), 97% (QC)
A. burnsii (sect. Alternaria) CBS 107.38 KP124124 97.1% (I), 97% (QC)
A. cerealis (sect. Alternaria) CBS 119544 KP124112 96.88% (I), 97% (QC)
A. arborescens (sect. Alternaria) EGS 39.128 KP275780 96.86% (I), 97% (QC)
OPA10-2 A. alternata (= A. citriarbusti, sect. Alternaria) SH-MIL-8s EF504044 99.21% (I), 100% (QC)
A. arborescens (sect. Alternaria) CBS 102605 KP124712 96.53% (I), 100% (QC)
A. arborescens (sect. Alternaria) EGS 39.128 EF504052 96.53% (I), 100% (QC)
A. alstroemeriae (sect. Alternaria) CBS 118809 KP124602 96.21% (I), 100% (QC)
  A. burnsii (sect. Alternaria) CBS 107.38 KP124734 96.05% (I), 99% (QC)
Chiang Mai J. Sci. 2021; 48(6) 1497

Figure 5. Drawing of morphological characteristics comparison of Alternaria species related to this

study, (a-c) redrawn from Simmons [44]. (d) redrawn from Elliott [12] and Nishikawa and Nakashima
[32]. (a) A. alternata; (b) A. arborescens; (c) A. gaisen; (d) A. iridicola. Bars: a-d 50 µm.

ACKNOWLEDGEMENTS
This work was supported by National Chiayi
University, Taiwan and the Mushroom Research
Foundation (MRF), Chiang Rai, Thailand; the
Thailand Research Fund and Mae Fah Luang
University entitled “Biodiversity, Phylogeny and
role of fungal endophytes on above parts of
Rhizophora apiculata and Nypa fruticans” (Grant
number: RSA5980068) for support. We would like
to thank Yi-Jyun Chen, Wu-Ting Tsai, Hsiu-Jung
Chien, Hsiang-Yu Lin, and Danushka S. Tennakoon
for general assistance. Sinang Hongsanan would
like to thank National Natural Science Foundation
of China (31950410548).
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