Original Article

Mycologia Iranica 11(1): 19-28, 2024 DOI: 10.22043/MI.2024.365383.1279

Three new Pleosporalean genera for the funga of Iran

A. Atashi Khalilabad ascomata, usually with a papillate apex, ostioles with

Kh.-B. Fotouhifar✉ or without periphyses, presence of cellular
Department of Plant Protection, Faculty of pseudoparaphyses, bitunicate asci, and ascospores of
Agriculture, College of Agriculture and Natural various shapes, pigmentations, and septations.
Resources, University of Tehran, Karaj, Iran Pleosporalean fungi are cosmopolitan and adapted to
various habitats, sometimes with extreme
Abstract: Pleosporalean fungi are important plant environmental conditions. In the recent years,
molecular studies coupled with morphological
pathogens, saprobes, and endophytes found in a wide
evidence have revealed numerous novel families,
range of economically important plants. To identify genera and species within the Pleosporales
the fungi associated with branch and stem canker (Wanasinghe et al., 2020; Lu et al., 2022;
symptoms in plants, the gardens and forests of Guilan Wanasinghe et al., 2022).
and Mazandaran provinces were surveyed, and The genus Ruta L. belonging to the Rutaceae
infected plant samples were collected from common family with common name of ''Iranian Sodab or
Sadab'', has two species in Iran; R. chalepensis L. and
hawthorn (Crataegus monogyna), common rue (Ruta
R. graveolens L. (Soleimani et al., 2009). Some
graveolens), and oriental persimmon (Diospyros kaki) fungal species have been reported from Ruta plants.
plants during the autumn of 2021. Fungal strains were Liberato and Barreto (2006) have reported Oidiopsis
isolated and purified by common procedures, and haplophylli (Magnus) Rulamort from Ruta graveolens
then were morphologically identified. Molecular in Brazil. Common hawthorn (Crataegus monogyna
identification of the fungal strains was performed Jacq.), a wild fruit tree, is one of the most interesting
using the sequence data of the ITS rDNA region. species of the Rosaceae family (Gundogdu et al.,
2014). This plant is commonly found as a shrub or
Based on the combined data, three fungal genera and
small tree, with small dark red fruit, commonly called
their related species belonging to the order haw (Sallabanks, 1992). The largest number of
Pleosporales including Acrocalymma walkeri from natural populations of this plant can be found in the
oriental persimmon, Setophaeosphaeria different regions of Iran (Mozafarian, 1996). A
badalingensis from common rue, and Tremateia number of fungi have been reported from common
chromolaenae from common hawthorn, were hawthorn. Montecchio et al. (2002) have reported
identified and characterized. All these three species Coniothyrium sporulosum (W. Gams & Domsch) Aa,
from C. monogyna in Italy. In an investigation,
are new to the funga of Iran. In addition, Diospyros
Gymnosporangium globosum (Farl.) Farl. was first
kaki, Crataegus monogyna, and Ruta graveolens have reported from American hawthorn by Yun et al.
been reported as new hosts (matrix nova) for the (2008) in South Korea. According to the surveys in
respective identified fungal taxa worldwide. Mexico, species of Alternaria Nees, Aureobasidium
Keywords: Ascomycota, Pleosporales, Morphology, Viala & G. Boyer, Drechslera S. Ito, Fusarium Link,
Phylogeny, ITS rDNA. Paecilomyces Bainier, and Ulocladium Preuss were
reported as the fungal community associated with
INTRODUCTION Crataegus sp. (Salazar-Cerezo et al., 2020). Also,
In the Dothideomycetes (Ascomycota), order Chen et al. (2022) investigated the associated fungus
Pleosporales that is commonly called as with hawthorn in China, and reported Diplocarpon
pleosporalean fungi, is the most species-rich and also mespilicola as new species. Several fungal species
is the largest order in this class, including more than such as Gymnosporangium confusum Plowr.,
10000 species. The Pleosporales, was first proposed Phyllactinia mali (Duby) U. Braun, Podosphaera
by Luttrell (1955) and formally established by Barr clandestine (Wallr.) Lév. and Rosellinia necatrix
(1987). This order is characterized by pseudothecioid

Submitted 6 April 2024, accepted for publication 6 May 2024

 Corresponding Author: E-mail: [email protected]
© 2024, Published by the Iranian Mycological Society
http://mij.areeo.ac.ir
 20 Mycologia Iranica - Vol. 11, 2024

Berl. ex Prill. have been reported from Crataegus contaminants. After surface disinfection by
monogyna in Iran (Ershad 2022). immersion of plant pieces in 70% ethanol for 2 min
The genus Diospyros L., (persimmon) of the and subsequently in 2% sodium hypochlorite
Ebenaceae family, has many species, including (NaClO) solution for 2 min, all plant pieces were
Japanese or oriental persimmon (Diospyros kaki L.), rinsed twice with sterile distilled water for 2 min.
which is a common edible cultivated species Then, disinfested plant pieces were dried between
(Mansoory et al. 2022). Like other plants, many fungi sterile paper towels and placed onto 2% water agar
have been reported from persimmon. In an (WA), and Petri dishes were kept at 25ºC under a 12
investigation, Cladosporium cladosporioides h photoperiod in the incubator for seven days. Fungal
(Fresen.) G.A. de Vries, was reported as sooty mold isolates were purified on potato dextrose agar (PDA)
fungus from D. kaki by Kwon and Park (2003) in culture medium using the hyphal tip method, and then
South Korea. Kwon et al. (2012), based on the the isolates were incubated at 25±1 °C until the pure
morphological features, pathogenicity tests, and fungal colonies were appeared. For long–term
molecular data, have identified Zygophiala storage, fungal isolates were grown on sterile filter
wisconsinensis Batzer & Crous (current name: papers placed on the PDA for 7–10 days.
Schizothyrium wisconsinense (Batzer & Crous) Crous Subsequently, colonized filter papers were taken from
& Batzer) on sweet persimmon in South Korea. the surface of culture medium, dried at room
Yamamoto et al. (2012) have isolated and temperature for four to five days, and then stored at -
characterized Adisciso kaki Kaz. Tanaka, J. Yamam. 20 °C for future use.
& Toy. Sato from D. kaki in Japan. Moyo et al. Morphology
(2016) have studied several fungi associated with The morphological characterization of the fungal
persimmon tree based on morphological features and isolates was performed based on the morphology of
molecular data. As a result, many fungal species such colony, as well as features of the fruiting bodies such
as Diaporthe infecunda R.R. Gomes, Glienke & as conidiomata (pycnidia), conidiogenous cells,
Crous, Diplodia mutila (Fr.) Mont., Eutypa lata conidia, ascomata, asci, and ascospores.
(Pers.) Tul. & C. Tul., Eutypella citricola Speg., Morphological studies of the three obtained genera
Phaeoacremonium parasiticum (Ajello, Georg & were performed on PDA, and the cultures were
C.J.K. Wang) W. Gams, Crous & M.J. Wingf., and incubated under near–ultraviolet (nUV) light (12 h
Neofusicoccum australe (Slippers, Crous & M.J. light/12 h darkness) at 25 °C. After 10–14 days
Wingf.) Crous, Slippers & A.J.L. Phillips were incubation of the pure fungal colonies, the fungal
isolated and reported from South Africa. Furthermore, features were assessed by light microscope using the
Asadi and Babaeizad (2016) have reported Alternaria microscopic slides mounts prepared in lacto–phenol
alternata (Fr.) Keissl. from persimmon tree in Iran. or lacto–phenol cotton blue solutions. Colony
Colletotrichum horii B.S. Weir & P.R. Johnst., was diameter of the fungal strains was measured usually
also isolated and reported from persimmon by Jeon et after 10 and 14 days. Macro– and micro–
al. (2017) in South Korea. Pestalotiopsis kaki K. Das, morphological features of the different recovered
S.Y. Lee & H.Y. Jung, was introduced as a novel isolates were measured. Micro–morphological
species from persimmon tree by Das et al. (2021) in features and measurements were performed according
South Korea. In Iran, several fungal species such as to Shoemaker et al. (1991), Crous et al. (2014) and
Ganoderma austral (Fr.) Pat., Pseudocercospora kaki Mapook et al. (2020). Photographs were taken using
Goh & W.H. Hsieh and Stereum hirsutum (Willd.) the BH2 light microscope (Olympus, Japan).
Pers. have been reported from D. kaki (Ershad 2022). Phylogeny
The main aim of the present study was the After morphological identifications, DNA was
morphological and phylogenetic identification and extracted from seven-day old fungal mycelium of the
characterization of the new fungal taxa belonging to recovered fungal strains using the method described
the pleosporalean fungi in Guilan and Mazandaran by Zhong and Steffenson (2001). The complete
provinces, Iran. internal transcribed spacer (ITS1-5.8S-ITS2) region
of rDNA was amplified using the ITS1
MATERIALS AND METHODS (TCCGTAGGTGAACCTGCGG) and ITS4
Fungal isolation (TCCTCGCTTATTGATATGC) primers (White et
Branches and stems with canker symptoms were al. 1990). PCR amplification carried out in a final
collected from common hawthorn (C. monogyna), volume of 25 μl containing 10 μL of Taq DNA
common rue (R. graveolens) and oriental persimmon polymerase Mix Red–Mgcl2 (Sinaclon, Iran), 11 μL
(D. kaki) plants from Guilan and Mazandaran deionized water, 1 μL of each primer (10 pmol) and 2
provinces during the autumn of 2021. Fungal strains μL of template DNA. The PCR amplification
were isolated from the diseased plant samples which performed in Eppendorf Thermal Cycler
bearing canker symptoms using the method described (Mastercycler, Germany) with an initial denaturation
by Refaei et al. (2011) with some minor at 94ºC for 3 min, followed by 35 cycles of
modifications. At first, diseased tissues were cut in denaturation step at 94ºC for 30 s, annealing at 60°C f
small pieces (1 cm2), and then washed in running tap or 30 s, and extension at 72°C for 30 s, and
water for 10 min to eliminate the surface terminated with a final extension step at 72°C for 10
 ATASHI KHALIILABAD AND FOTOUHIFAR : Three new Pleosporalean genera for the funga of Iran 21

min The PCR product was analyzed in 1.5% agarose (Kumar et al. 2018). Bootstrap analysis (Felsenstein
gel by gel electrophoresis technique with 1x Tris- 1985) of the ML tree was performed with 1000
Boric acid-EDTA buffer (TBE) and finally PCR replicates (Fig. 2). Sequences generated in the current
products were sent to Cardiogenetic Research Center study were deposited in the GenBank. Detailed
(Tehran, Iran) for sequencing. information of the examined sequences in this study
Newly obtained DNA sequences were manually are provided in Table 1.
edited with Chromas 2.6.6 software (Technelysium,
RESULTS
Australia) and the edited sequences were saved in
FASTA format. The resulting sequences (450-620 bp) In the present study, three species belonging to three
were subjected to BLAST search (Altschul et al. different pleosporalean genera including
1990) to find the most similar sequences in the Acrocalymma walkeri, Setophaeosphaeria
National Center for Biotechnology Information badalingensis, and Tremateia chromolaenae were
(NCBI) (www.ncbi.nlm.nih. gov/genbank/). Thirty- obtained and characterized based on both
eight reference ITS sequences of Setophaeosphaeria, morphological criteria and molecular data. All three
Acrocalymma, Tremateia species as well as the ITS genera are new taxa for the funga of Iran.
sequences of Sarcinomyces crustaceus (AJ244258) Furthermore, Diospyros kaki, Ruta graveolens, and
and Daldinia concentrica (JX658475) as outgroup Crataegus monogyna are reported as new hosts
were selected for the phylogenetic analyses (Table 1). (matrix nova) for the respective recovered fungal
Then, the sequences were aligned with Clustal W species worldwide.
(Thompson et al. 1994). Maximum likelihood (ML)
analysis (Felsenstein 1973) of the aligned sequences
was performed by heuristic search with Mega X

Table 1. ITS rDNA sequences used for phylogenetic analyses. Newly generated sequences are in boldface.
GenBank accession
Collection Host/Substrate no.
Species Origin Reference
number/Strain
ITS
Setophaeosphaeria hemerocallidis CBS 138006 Hemerocallis fulva China NR171715 Crous et al. (2014)
CBS 138006 Hemerocallis fulva China KJ869161 Crous et al. (2014)
Strain BRPET38 Broussonetia papyrifera China MT658111 Xu (2020)
S. badalingensis ABRIICC 10360 Ruta graveolens Iran ON544079 This study
CBS 138007 Hemerocallis fulva China NR171716 Crous et al. (2014)
CBS 138007 Hemerocallis fulva China KJ869162 Crous et al. (2014)
S. sidae CBS 135108 Sida sp. Brazil NR156261 Quaedvlieg et al. (2013)
CBS 135108 Sida sp. Brazil KF251149 Quaedvlieg et al. (2013)
S. citricola KACC 49591 Amaranthus patulus South Korea MW412749 Choi (2020)
CBS 143179 Citrus australasica Australia MH107916 Crous et al. (2018)
S. citri Strain CPC 27148 Citrus reticulata Italy MG263524 Guarnaccia and Crous (2017)
S. microspora - Soil China MK329132 Zhang et al. (2021)
CGMCC 3.19301 Soil China NR172843 Zhang and Cai (2018)
- Soil China MK329131 Zhang and Cai (2018)
Acrocalymma pterocarpi MFLUCC 17-0926 Pterocarpus indicus Thailand NR163327 Jayasiri (2018)
Voucher C233 Pterocarpus indicus Thailand MK347732 Jayasiri et al. (2019)
A. medicaginis Yeh 0049 Ipomoea pes-caprae Taiwan MW376531 Yeh (2020)
CPC 24340 Medicago sativa Australia KP170620 Trakunyingcharoen et al. (2014)
A. ampeli NCYU 19-0008 Ficus ampelas Taiwan MW063151 Tennakoon et al. (2021)
MFLU 19-2734 Ficus ampelas Taiwan MW063150 Tennakoon et al. (2021)
A. walkeri ABRIICC 10353 Diospyros kaki Iran OL376691 This study
CBS 257.93 - Australia MH862398 Vu et al. (2019)
A. vagum Strain 1186 Nervilia fordii China MZ400559 Tan et al. (2021)
Strain DSE1 Glycyrrhiza uralensis China MW042345 Li (2020)
A. fici NTOU 4481 - Taiwan MZ422889 Cha et al. (2021)
BR68 Calamus castaneus Malaysia MN637807 Azuddin and Zakaria (2019)
A. aquatica MFLUCC 11-0208 - China NR121544 Schoch et al. (2014)
MFLUCC11-0208 - China JX276951 Zhang et al. (2012)
Tremateia chiangraiensis MFLUCC 17-1428 Chromolaena odorata Thailand NR168867 Mapook (2020)
MFLUCC 17-1429 Chromolaena odorata Thailand MT214356 Mapook (2020)
T. chromolaenae ABRIICC 10342 Crataegus monogyna Iran MZ226451 This study
MFLUCC 17-1425 Chromolaena odorata Thailand NR168868 Mapook (2020)
T. thailandensis MFLUCC 17-1430 Chromolaena odorata Thailand NR168869 Mapook (2020)
MFLUCC 17-1430 Chromolaena odorata Thailand MT214361 Mapook et al. (2020)
T. murispora HKAS 104642 Decaying wood China NR165916 Feng (2019)
GZCC 18-2787 Decaying wood China MK962245 Feng et al. (2019)
T. camporesii MFLU 19-2109 Dead branch Thailand NR169985 Samarakoon and Hyde (2019)
MFLU 19-2109 Dead branch Thailand MN473061 Samarakoon and Hyde (2020)
Sarcinomyces crustaceus CBS 156.89 Endocronartium Netherlands AJ244258 de Hoog et al. (1999)
harknessii gall
Daldinia concentrica CBS 117124 Platanus sp. Greece JX658475 Stadler et al. (2013)
 22 Mycologia Iranica - Vol. 11, 2024

Molecular phylogeny Acrocalymma walkeri provided by Shoemaker et al.

As shown in Fig. 1, phylogenetic analyses and (1991). In the phylogenetic tree, our Acrocalymma
obtained tree topology revealed that the obtained walkeri isolate (OL376691) was grouped with other
isolates from Iran are closely related to the relevant species of the genus in the clade II with 98%
sequences which were deposited in the GenBank and bootstrap support (Fig. 1). Among the 18 reported
placed in three distinct clades from top to the bottom Acrocalymma species, only A. arengae Konta & K.D.
of the phylogenetic tree, corresponding to fungal Hyde, A. pterocarpi Jayasiri, E.B.G. Jones & K.D.
families of Phaeosphaeriaceae (Clade I), Hyde, A. chuxiongense Y.W. Liu & X.Y. Zeng, A.
Acrocalymmaceae (Clade II), and hongheense Mortimer, and A. walkeri have known
Didymosphaeriaceae (Clade III) of the Pleosporales teleomorph, and the other 13 species have only
(class Dothideomycetes) in the Ascomycota. species anamorphic state. Morphological difference of A.
identified in this study can be distinguished from walkeri with other related species is that the A.
other reported species according to their substrate walkeri has bigger ascospores compared to A.
(host plant). Setophaeosphaeria species were placed pterocarpi and smaller ascospores compared to A.
in Clade I of the phylogenetic tree, with 80% arengae, A. chuxiongense and A. hongheense (Konta
bootstrap support. The S. badalingensis isolate et al. 2023, Liu & Zeng. 2022).
(accession no. ON544079) was 99% identical to that Setophaeosphaeria badalingensis Crous & Y. Zhang
of other isolates of this species from GenBank ter, Persoonia 32: 271 (2014)
(NR171716 and KJ869162). Acrocalymma species Colony on PDA slow growing, attaining 29 mm in
were situated in Clade II of the phylogenetic tree, diam. after 10 days at 25 °C under near ultra violet
with 98% bootstrap support. The A. walkeri isolate (nUV) light, 12 h light/12 h dark. Colony from above:
(accession no. OL376691) was 100% identical to that fluffy to velvety at the center and covered by floccose
of another isolate from GenBank (MH862398). aerial mycelia, circular, with an entire smooth edge,
Different Tremateia species were placed in Clade III, initially white, becoming smoke grey on surface;
with 100% bootstrap support. The partial ITS rDNA reverse: olivaceous grey to dark grey with distinct
sequence of the recovered T. chromolaenae isolate sectors. Conidiomata pycnidial, covered with hyphae,
(accession no. MZ226451) was 100% identical to that 220–310 × 210–300 μm (x̄ = 250 × 260 μm, n = 20),
of another isolate from GenBank (NR168868). aggregated or solitary, pale brown with dark central
Taxonomy ostiole. Ostiole surrounded by brown, unbranched,
Acrocalymma walkeri (Shoemaker, C.E. Babcock & thick-walled, septate, verruculose setae. Textura
J.A.G. Irwin) Crous & Trakun., IMA Fungus 5 (2): angularis, pale brown. Conidiophores absent.
407 (2014) Conidiogenous cells, hyaline, smooth, ampulliform to
Colony on PDA attaining 46 mm in diam. after 14 doliiform, 5–7 × 3.5–4.5 μm (x̄ = 6 × 3/5 μm, n = 20),
days at 25 °C under near ultra violet (nUV) light, 12 h phialidic with prominent periclinal thickening.
light/12 h dark. Colony from above: medium dense, Conidia solitary, hyaline, aseptate, smooth, two
circular, with an entire smooth edge, slightly raised, biguttulate to multiguttulate, subcylindrical with
fluffy to velvety, olivaceous to grayish at the margin, obtuse ends, straight or gently curved, 4.5–7 × 2.5–
white in the center; reverse: white at the margin, dark 3.5 μm (x̄ = 6 × 3 μm, n = 50). Sexual morph: not
brown to black in the center. Sexual morph: observed (Fig. 3).
Ascomata 180–280 × 170–270 μm (x̄ = 220 × 210 Specimen examined. Iran, Gilan province, Asalem, N
μm, n = 20), solitary or aggregated, immersed to 37°43'52.5" E 48°57'38.0", recovered from stem of
semi-immersed, dark brown, globose, without ostiole, Ruta graveolens with canker symptom, November
wall of textura angularis, with brown to pale brown 2021, Abbas Atashi Khalilabad, isolate UT2021,
cells. Asci 55–85 × 8–12 μm (x̄ = 75 × 10 μm, n = GenBank Accession No. ON544079, ABRIICC
30), hyaline, bitunicate, fissitunicate, cylindrical, with 10360.
a short stalk having eight overlapping biseriate Note: Morphological features of the investigated
ascospores. Ascospores 17–23 × 5–6 μm (x̄ = 20 × 5 isolate were similar to the description of
μm, n = 50), overlapping, obliquely biseriate Setophaeosphaeria badalingensis provided by Crous
overlapping, hyaline, fusiform, first with 1 -septate, et al. (2014). Phylogenetic analyses indicated that S.
becoming 3-septate when mature, with four guttules, badalingensis is a sister species to S. hemerocallidis
smooth, narrowly rounded ends, constricted at median Crous & Y. Zhang ter (CBS 138006 and BRPET38)
septum. Asexual morph: not observed (Fig. 2). with 76% bootstrap support (Fig. 1). However,
Specimen examined. Iran, Mazandaran province, morphologically, S. hemerocallidis differs from S.
Babol, N 36°32'60.0" E 52°40'60.0, recovered from badalingensis in having bigger conidia (11–)13–16(–
branch of Diospyros kaki with canker symptom, 19) × (3–)3.5(–4) µm (Crous et al. 2014).
November 2021, Abbas Atashi Khalilabad, isolate Tremateia chromolaenae Mapook & K.D. Hyde,
UZB-RB, GenBank Accession No. OL376691, Fungal Diversity 101: 40 (2020)
ABRIICC 10353. Colony on PDA attaining 54 mm in diam. after 14
Note: Morphological features of the investigated days at 25 °C under near ultra violet (nUV) light, 12 h
isolate were similar to the description of light/12 h dark. Colony from above: circular, with
 ATASHI KHALIILABAD AND FOTOUHIFAR : Three new Pleosporalean genera for the funga of Iran 23

smooth margin, mycelium slightly raised, initially clavate, with eight ascospores, straight or slightly
white, becoming pale pinkish white on surface; curved, apically rounded, pedicellate. Ascospores 17–
reverse: brown to pale brown, white to creamy white 21 × 6–9 μm (x̄ = 19 × 7.8 μm, n = 50), overlapping
at margins. Sexual morph: Ascomata 150–225 × 160– uni- two seriate, initially hyaline, becoming golden-
220 μm diam (x̄ = 195 × 188 μm, n = 10), immersed, brown at maturity, muriform, oval, ellipsoidal to
solitary or aggregated, pale to dark brown to brown, subfusiform, straight or slightly curved, 3–6
globose, with ostiolar protruding neck, arranged in a transversely septate, with 1–2 vertical septa,
textura angularis with brown to pale brown cells, Asci constricted at the central septum, surrounded by a
75–100 × 14–19 μm (x̄ = 88 × 17.5 μm, n = 20), distinct hyaline gelatinous sheath. Asexual morph:
hyaline, bitunicate, fissitunicate, clavate to cylindric- not observed (Fig. 4).

Fig. 1. Maximum likelihood (ML) tree generated in MEGA X, based on aligned sequences of the ITS rDNA
regions of 38 isolates of Setophaeosphaeria, Acrocalymma, Tremateia species, and Sarcinomyces crustaceus
(AJ244258) and Daldinia concentrica (JX658475) as the out-group sequences. Bootstrap values (1000 replicates) are
indicated at the nodes. Sequences generated in the current study are in boldface. The scale bar indicates 0.10
expected nucleotide changes per site.
 24 Mycologia Iranica - Vol. 11, 2024

Fig. 2. Acrocalymma walkeri isolate UZB-RB. A.-B. Colony obverse and reverse on PDA incubated for 14 d at
25 °C in 12/12 h dark/nUV condition, C.-D. Ascomata, E.-F. Asci, and G. Ascospores. Scale bars: C = 200 μm. D=
50 μm. E–G = 10 μm.

Fig. 3. Setophaeosphaeria badalingensis, isolate UT2021. A.-B. Colony obverse and reverse on PDA incubated
for seven d at 25 °C in 12/12 h dark/nUV condition, C. Colony obverse on PDA after 14 d at 25 °C in 12/12 h
dark/nUV condition, D. Pycnidium with setae, E.-F. Conidiogenous cells, and G. Conidia. Scale bars: D = 100 μm.
E–G = 5 μm.
 ATASHI KHALIILABAD AND FOTOUHIFAR : Three new Pleosporalean genera for the funga of Iran 25

Specimen examined. Iran, Gilan province, Asalem, N the isolates were grouped into three categories. Based
37°44'03.8"E 48°57'07.9", recovered from branch of on morphological and molecular analyses, these three
Crataegus monogyna with canker symptom, isolates were identified as Acrocalymma walkeri,
November 2021, Abbas Atashi Khalilabad, isolates Setophaeosphaeria badalingensis, and Tremateia
UTFS-17, GenBank Accession No. MZ226451, chromolaenae in Diospyros kaki (oriental
ABRIICC 10342. persimmon), Ruta graveolens (common rue), and
Note: Morphological features of the investigated Crataegus monogyna (common hawthorn),
isolate were similar to the description of Tremateia respectively. Isolate UT2021 was identified as S.
chromolaenae provided by Mapook et al. (2020). In badalingensis based on both the morphological
the phylogenetic analyses, sequences of five description provided by Crous et al. (2014) and
Tremateia species were used. Our isolate (ABRIICC molecular data. Until 2022, only seven species of
10342) clustered with another isolate of T. Setophaeosphaeria have been identified and
chromolaenae (NR168868) with maximum bootstrap described including S .badalingensis ,S .citri Guarn.
support of 100% (Figure. 1). Tremateia & Crous ,S .citricola Crous & M.J. Wingf ,.S .
chromolaenae differs from T. thailandensis Mapook hemerocallidis ,S .microspora Z.F. Zhang & L. Cai ,
& K.D. Hyde, one of the nearest species, in having S .setosa (Leuchtm.) Crous and S .sidae (Quaedvl.,
smaller ascomata, asci and ascospores (Mapooket al. Verkley, R.W. Barreto & Crous) Crous
2020). (https://www.mycobank.org). Setophaeosphaeria
badalingensis can be distinguished from other species
DISCUSSION
of the genus based on the nucleotide sequence of the
In this study, three fungal isolates were obtained from ITS region.
the stems and branches of growing plants with canker
symptoms in Guilan and Mazandaran, Iran. After
examination of cultural and morphological features,

Fig. 4. Tremateia chromolaenae isolate UTFS-17. A.-B. Colony obverse and reverse on PDA incubated for 14 d at
25 °C in 12/12 h dark/nUV condition, C.-D. Ascomata, E.-F. Asci, and G. Ascospores. Scale bars: C = 60 μm. D= 30
μm. E–G = 20 μm.
 26 Mycologia Iranica - Vol. 11, 2024

This species was previously reported from with Entomosporium leaf spot on hawthorn in
Hemerocallis fulva (L.) L. (Crous et al. 2014). This is China. Plant Disease 106: 2884-2891.
the first report of S. badalingensis as a new taxon for Crous, P.W., Shivas, R.G., Quaedvlieg, W.V., Van
the funga of Iran. In addition, S. badalingensis is der Bank, M. et al. 2014. Fungal Planet
reported here for the first time as a fungus associated description sheets: 214–280. Persoonia 32: 184-
with stem canker symptoms in R. graveolens. Isolate 306.
UZB-RB was identified as Acrocalymma walkeri Das, K., Lee, S.Y. and Jung, H.Y. 2021.
based on the description provided by Shoemaker et al. Pestalotiopsis kaki sp. nov., a novel species
(1991) as well as molecular data (Vu et al. 2019). isolated from persimmon tree (Diospyros kaki)
Acrocalymma walkeri was originally described by bark in Korea. Mycobiology 49: 54-60.
Shoemaker et al. (1991) as Massarina walkeri de Hoog, G. D., Zalar, P., Urzi, C., De Leo, F.,
Shoemaker, C.E. Babc. & J.A.G. Irwin with Yurlova, N. A., and Sterflinger, K. 1999.
Acrocalymma medicaginis as its an asexual morph of Relationships of Dothideaceous black yeasts and
this fungus. A recent phylogenetic analysis by meristematic fungi based on 5.8 S and ITS2
Trakunyingcharoen et al. (2014) based on the rDNA sequence comparison. Studies in Mycology
sequence of the ITS rDNA region revealed that M. 43: 31-37.
walkeri is nestled inside the broader Acrocalymma Ershad, D. 2022. Fungi and fungal analogues of Iran.
lineage. Therefore, Trakunyingcharoen et al. (2014) Iranian Research Institute of Plant Protection,
transferred M. walkeri to the genus Acrocalymma and Agricultural Research, Education and Extension
treated A. medicaginis and A. walkeri as distinct Organization, Tehran, Iran, 712 pp.
species. This species has been previously reported in Felsenstein, J. 1973. Maximum likelihood and
Medicago sativa (Trakunyingcharoen et al. 2014). minimum-steps methods for estimating
This is the first report of A. walkeri as a new taxon for evolutionary trees from data on discrete
the funga of Iran. In addition, A. walkeri was reported characters. Systematic Biology 22: 240-249.
for the first time as a fungus associated with branch Felsenstein, J. 1985. Confidence limits on
canker symptoms in D. kaki trees worldwide. The phylogenies: an approach using the bootstrap.
isolate UTFS-17 was identified as Tremateia Evolution 39: 783-791.
chromolaenae based on the description provided by Gundogdu, M., Ozrenk, K., Ercisli, S., Kan, T.,
Mapook et al. (2020) as well as molecular data. Kodad, O. and Hegedus, A. 2014. Organic acids,
Tremateia chromolaenae differs from other species of sugars, vitamin C content and some pomological
this genus based on its smaller ascomata. This species characteristics of eleven hawthorn species
was previously reported as a saprobic fungus from (Crataegus spp.) from Turkey. Biological
Chromolaena odorata (Mapook et al. 2020). This is Research 47: 1-5.
the first report of T. chromolaenae as a new taxon for Jeon, J.Y., Hassan, O., Chang, T., Lee, D.W., Shin,
the funga of Iran. In addition, T. chromolaenae is J.S. and Oh, N.K. 2017. Anthracnose of
reported for the first time as a fungus associated with persimmon (Diospyros kaki) caused by
branch canker symptoms in C. monogyna trees Colletotrichum horii in Sangju, Korea. Plant
worldwide. Disease 101: 1035-1035.
Konta, S., Tibpromma, S., Karunarathna, S. C.,
ACKNOWLEDGMENTS
Samarakoon, M. C., Steven, L. S., Mapook, A.,
We are thankful to the University of Tehran for and Hyde, K. D. 2023. Morphology and multigene
supporting this research. phylogeny reveal ten novel taxa in Ascomycota
from terrestrial palm substrates (Arecaceae) in
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 ‫‪28‬‬ ‫‪Mycologia Iranica - Vol. 11, 2024‬‬

‫سه جنس جدید از راسته ‪ Pleosporales‬برای فونگای ایران‬

‫✉‬
‫عباس آتشی خلیل آباد و خلیلبِردی فتوحی فر‬
‫گروه گیاهپزشکی‪ ،‬دانشکده کشاورزی‪ ،‬دانشکدگان کشاورزی و منابع طبیعی‪ ،‬دانشگاه تهران‪ ،‬کرج‪ ،‬ایران‬

‫چکیده‪ :‬قارچ های متعلق به راسته ‪ )Pleosporalean( Pleosporales‬بیمارگرهای گیاهی‪ ،‬پودهزی و یا اندوفیت طیف وسیعی از‬
‫گیاهان مهم از نظر اقتصادی هستند‪ .‬به منظور شناسایی قارچهای همراه با عالئم شانکر شاخه و ساقه در گیاهان‪ ،‬در پاییز ‪1400‬‬
‫باغات و جنگلهای استانهای گیالن و مازندران مورد بازدید قرار گرفت و نمونههای گیاهی دارای عالئم از درخت زالزالک معمولی‬
‫(‪ ،)Crataegus monogyna Jacq.‬بوته سداب معمولی (‪ )Ruta graveolens L.‬و درخت خرمالوی شرقی (‪)Diospyros kaki L.‬‬
‫جمعآوری شدند‪ .‬استرینهای قارچی با روش های معمول جداسازی و خالص سازی شدند و بر اساس خصوصیات ریخت شناختی‬
‫شناسایی شدند‪ .‬در نهایت‪ ،‬شناسایی مولکولی استرین های قارچی هم با استفاده از توالی یابی ناحیه ‪ ITS rDNA‬انجام شد‪ .‬بر‬
‫اساس تلفیق دادهها‪ ،‬در نهایت سه جنس و گونه قارچی متعلق به راسته ‪ Pleosporales‬شامل گونه ‪ Acrocalymma walker‬از‬
‫خرمالو شرقی‪ ،‬گونه ‪ Setophaeosphaeria badalingensis‬از سداب معمولی و گونه ‪ Tremateia chromolaenae‬از زالزالک معمولی‬
‫شناسایی و توصیف شدند‪ .‬در تحقیق حاضر‪ ،‬هر سه گونه قارچی برای فونگای ایران جدید هستند‪ .‬همچنین گیاهان خرمالوی‬
‫شرقی‪ ،‬زالزالک معمولی و سداب معمولی به عنوان میزبانهای جدید (‪ )matrix nova‬برای گونههای قارچی مورد بررسی در جهان‬
‫گزارش می شوند‪.‬‬
‫کلمات کلیدی‪ :‬آسکومیکوتا‪ ،Pleosporales ،‬ریخت شناسی‪ ،‬فیلوژنی‪ITS rDNA ،‬‬

‫مکاتبه کننده‪ :‬خلیلبِردی فتوحی فر ‪Email: [email protected]‬‬

‫تاریخ دریافت‪ 1403/1/18 :‬تاریخ پذیرش‪1403/2/17 :‬‬