Research Article
Print
Research Article
Two new pestalotioid fungi from tropical fruits in Iran
expand article infoAmirreza Amirmijani‡, Adel Pordel§, Kowsar Dehghani‡, Mohammad Javad Pourmoghaddam|, Hossein Masigol, Hans-Peter Grossart#
‡ University of Jiroft, Jiroft, Iran
§ Plant Protection Research Department, Baluchestan Agricultural and Natural Resources Research and Education Center, Iranshahr, Iran
| University of Guilan, Rasht, Iran
¶ Department of Plankton and Microbial Ecology, Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB), Stechlin, Germany
# Potsdam University, Potsdam, Germany

Corresponding author: Amirreza Amirmijani ( ar.amirmijani@ujiroft.ac.ir )

Academic editor: Ning Jiang
© 2025 Amirreza Amirmijani, Adel Pordel, Kowsar Dehghani, Mohammad Javad Pourmoghaddam, Hossein Masigol, Hans-Peter Grossart.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Amirmijani A, Pordel A, Dehghani K, Pourmoghaddam MJ, Masigol H, Grossart H-P (2025) Two new pestalotioid fungi from tropical fruits in Iran. MycoKeys 115: 221-240. https://doi.org/10.3897/mycokeys.115.136469
Open Access

Abstract

In a survey of tropical plant diseases in southern and southeastern Iran, samples of diseased Mangifera indica and Psidium guava leaves with necrotic symptoms were collected between 2021 and 2022. Six representative isolates of Neopestalotiopsis and Robillarda (three isolates for each) were studied using morphological characteristics as well as multi-locus phylogenetic analysis based on (i) the internal transcribed spacer (ITS) region of the nuclear rDNA, (ii) part of the translation elongation factor 1-alpha (tef1), and (iii) the β-tubulin (tub2). After morphological investigation, our phylogenetic analysis revealed that the Neopestalotiopsis and Robillarda isolates under study differed from all previously described species within these genera. Based on our polyphasic approach, two new species, including Neopestalotiopsis guava sp. nov. from necrotic Mangifera indica and Robillarda khodaparastii sp. nov. from Psidium guava are described and illustrated from Iran.

Key words:

Fungal diversity, Mangifera indica, Pestalotioid fungi, Psidium guava, Tropical fruits, two new species

Introduction

Tropical fruits provide essential nutrition and serve as a source of income for farmers engaged in export production. The major cultivation areas for tropical fruits are predominantly in developing countries, particularly in Asia and Latin America (FAO 2020). Global trade in tropical fruits has expanded to unprecedented levels in recent years, reaching an aggregate export volume of nearly 9 million tons in 2023. Strong demand from key importing countries has driven significant investments in productivity improvements and expansion of cultivation areas in supplying countries, notably for avocados (FAO 2024). Iran is a major producer of fruits, including tropical fruits, due to favorable meteorological conditions, diverse climates, and significant temperature differences between the northern and southern regions (Saboki et al. 2012). Regarding tropical fruit production, the Provinces of Hormozgan and Sistan and Baluchistan, located in the south and southeast of Iran, are considered the most suitable regions due to their proximity to the equator, the Oman Sea, and the Indian Ocean (Saboki et al. 2012, 2014).

Pestalotioid fungi are diverse and exhibit various lifestyles, including plant pathogens, endophytes, and saprophytes (Crous et al. 2015a; Sun et al. 2023). Despite uncertainties regarding the generic divisions among pestalotioid fungi, the classification system based on the number of conidia cells in different genera is still in use (Shu et al. 2020). The genus Pestalotia was initially described by De Notaris in 1841 (Jiang et al. 2022). Subsequently, based on the number of conidia cells, Pestalotia was divided into four distinct genera: Truncatella (4 cells), Pestalotiopsis (5 cells), Pestalotia (6 cells), and Monochaetia (5 cells) (Sutton 1980). Phylogenetic analyses have validated Sutton’s classification based on morphological characteristics; however, species previously classified under Pestalotia have been reassigned to the genus Seiridium (Marin-Felix et al. 2019). Additionally, the genus Pestalotiopsis has been further categorized into Pestalotiopsis sensu stricto, Neopestalotiopsis, and Pseudopestalotiopsis (Maharachchikumbura et al. 2016). Neopestalotiopsis is distinct from Pestalotiopsis, characterized by the presence of versicolourous median cells (Jiang et al. 2022).

The genus Robillarda was introduced by Saccardo in 1880 to accommodate the type species R. sessile (Saccardo 1880). Robillarda is a pestalotioid fungus within the family Sporocadaceae, and characterized by conidia with appendage-bearing (Liu et al. 2019). Although the genus includes 41 morphological species, sequence data are available for only some of them (Liu et al. 2019; https://www.indexfungorum.org/names/Names.asp/, accessed 31 Aug 2024).

In the present study, leaf spots on Mangifera and Psidium were observed in Sistan and Baluchestan Province, Iran. Based on modern taxonomic approaches, we identified two new species of Neopestalotiopsis and Robillarda from Iran. Detailed morphological descriptions, illustrations, and phylogenetic information are provided here.

Materials and methods

Sampling and fungal isolation

During a survey conducted on tropical and subtropical fruit trees in the summer of 2021, a total of seventy-five samples were collected from mango (M. indica) plants displaying symptoms of leaf spot disease. The leaf samples were specifically gathered from various districts in the Provinces of Hormozgan (Siaho district) and Sistan and Baluchestan (Nikshahr, Ghasreghand, Rask, and Konarak districts), which are located in the southern and southeast regions of Iran, respectively. The infected samples were transported to the laboratory and stored in a refrigerator under dry conditions at a temperature of 4 °C. To begin the isolation process, the infected tissues were cut into 7–8 mm pieces, surface–disinfected with a 2% sodium hypochlorite solution for 3 min, rinsed twice with sterile distilled water, dehydrated, and subsequently placed on 2% water-agar (2% WA) in Petri dishes. These Petri dishes were maintained at 25 °C under alternating near-UV light and dark conditions (12 h light/12 h dark) for 7 days. After 48 h, conidia were observed growing on the leaf pieces and transferred to 2% WA using the single-spore method. Hyphal tips emerging from individual conidia were further transferred to a potato dextrose agar (PDA) medium to establish pure cultures (Refaei et al. 2011).

Morphological characterization

Mycelia plugs were extracted from the purified colony and placed on PDA to assess the colony’s overall characteristics. Subsequently, the plugs were incubated at 25 °C under alternating near-UV light and dark conditions. After 7–10 d, the color of the colony and the conidial mass were documented. To further analyze the morphological features of the conidiomata and conidia, more than 200 conidiophores and conidia were examined using slide mounts prepared with lactophenol and lactophenol cotton blue. Morphometrical analyses were also conducted on 200 conidiophores and conidia. For this purpose, a BH2 Olympus light microscope (Japan) equipped with a Microbin 12MP USB2.0 camera was utilized. The holotype and ex-type specimens have been deposited in the Herbarium of the Mycology Laboratory at the University of Jiroft, Jiroft, Iran (UJFCC).

DNA extraction, PCR, and sequencing

DNA was extracted from seven-day-old fungal mycelium using the protocol described by Zhong and Steffenson (2001). The entire internal transcribed spacer (ITS1-5.8S-ITS2) regions of the rDNA, the partial translation elongation factor 1- alpha (tef1) gene and b-tubulin (tub2) gene were amplified using the primer pairs ITS1 (5”-TCCGTAGGTGAACCTGCGG-3”) and ITS4 (5”-TCCTCCGCTTATTGATATGC-3”) (White et al. 1990), EF1-728F (5”-CATCGAGAAGTTCGAGAAGG-3”) and EF2 (5”-GGARGTACCAGTSATCATGTT-3”) (Carbone and Kohn 1999; O’Donnell et al. 1998), as well as Bt2a (5”-GGTAACCAAATCGGTGCTGCTTTC-3”) and Bt2b (5”-ACCCTCAGTGTAGTGACCCTTGGC-3”) (Glass and Donaldson 1995). PCR amplifications were carried out in a final volume of 25 μL. The PCR mixtures contained 10 μL of master mix (CinnaGen, Iran), which included 10 × PCR buffer, MgCl2, dNTPs, Taq DNA Polymerase, 11 μL of double-distilled water, 1 μL of each forward and reverse primers (10 pmol), and 2 μL of template DNA. The PCR amplifications were done using a thermocycler with the following thermal conditions for ITS: initial denaturation at 94 °C for 3 min, followed by 35 cycles of denaturation step at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, and terminated with a final extension step at 72 °C for 10 min; for tef1: initial denaturation at 94 °C for 8 min, and then followed by 35 cycles each with denaturation at 94 °C for 15 sec, annealing at 55 °C for 20 sec and the extension at 72 °C for 1 min, and a final extension at 72 °C for 5 min; for tub2: initial denaturation at 94 °C for 3 min, and then followed by 35 cycles each with denaturation at 95 °C for 30 s, annealing at 53 °C for 30 s and the extension at 72 °C for 45 s, and a final extension at 72 °C for 90 s. All amplicons were sent to the Codon Genetic Group (Tehran, Iran) for sequencing.

Phylogenetic analyses

To identify closely related taxa, BLASTn searches were done separately for the three loci. Type and reference sequences of related taxa were retrieved from the National Center for Biotechnology Information (NCBI), if available, based on recent publications on the genera Neopestalotiopsis (Maharachchikumbura et al. 2012, 2014a, 2014b; Fiorenza et al. 2022; Razaghi et al. 2024) and Robillarda (Crous et al. 2015a; Liu et al. 2019). All alignments were produced using the server versions of MAFFT v. 7.490 (http://mafft.cbrc.jp/alignment/server/; Katoh et al. 2019) and were manually checked and refined with MEGA Ver. 7 (Kumar et al. 2016). Following the results of BLASTn searches for generated sequences of the three loci (ITS, tef1, tub2), a phylogenetic analysis was performed for Neopestalotiopsis species including 91 isolates. Similarly, a phylogenetic placement was conducted for Robillarda, including 11 isolates. Pestalotiopsis colombiensis and P. diversiseta were selected as the outgroup taxa for both trees (Table 1). After excluding ambiguously aligned and gappy regions, the resulting combined data matrix contained 1363 alignment positions across all three loci (494 from ITS, 471 from tef1, and 398 from tub2) for Neopestalotiopsis and 1306 alignment positions (518 from ITS, 443 from tef1, and 345 from tub2) for Robillarda.

Table 1.
Download as
CSV
XLSX

Isolation and accession numbers of sequences used in the phylogenetic analyses. Isolates/sequences in bold were isolated/sequenced in present study. N/A: not available. 1 T indicates ex-type material.

Species Strain1 Origin GenBank accession numbers References
ITS tef1 tub2
Neopestalotiopsis acrostichi MFLUCC 17-1754T Thailand MK764272 MK764316 MK764338 Norphanphoun et al. (2019)
Neopestalotiopsis alpapicalis MFLUCC 17-2544T Thailand MK357772 MK463547 MK463545 Kumar et al. (2019)
Neopestalotiopsis aotearoa CBS 367.54T New Zealand KM199369 KM199526 KM199454 Maharachchikumbura et al. (2012)
Neopestalotiopsis asiatica MFLUCC 12-0286T China JX398983 JX399049 JX399018 Maharachchikumbura et al. (2012)
Neopestalotiopsis australis CBS 114159T Australia KM199348 KM199537 KM199432 Maharachchikumbura et al.(2014b)
Neopestalotiopsis brachiate MFLUCC 17-1555T Thailand MK764274 MK764318 MK764340 Norphanphoun et al. (2019)
Neopestalotiopsis brasiliensis COAD 2166T Brazil MG686469 MG692402 MG692400 Bezerra et al. (2018)
Neopestalotiopsis cavernicola KUMCC 20-0269T China MW545802 MW550735 MW557596 Liu et al. (2021)
Neopestalotiopsis celtidis CGMCC 3.23513T China OR247900 OR361449 OR381049 Razaghi et al. (2024)
Neopestalotiopsis chrysea MFLUCC 12-0261T China JX398985 JX399051 JX399020 Maharachchikumbura et al. (2012)
Neopestalotiopsis clavispora MFLUCC 12-0281T China JX398979 JX399045 JX399014 Maharachchikumbura et al. (2012)
Neopestalotiopsis coffeae-arabicae HGUP4019T China KF412649 KF412646 KF412643 Song et al. (2013)
Neopestalotiopsis concentrica CFCC 55162T China OK560707 OM622433 OM117698 Peng et al. (2022)
Neopestalotiopsis cubana CBS 600.96T Cuba KM199347 KM199521 KM199438 Maharachchikumbura et al.(2014b)
Neopestalotiopsis dendrobii MFLUCC 14-0106T Thailand MK993571 MK975829 MK975835 Ma et al. (2019)
Neopestalotiopsis dolichoconidiophora CGMCC 3.23490T China OR247911 OR361421 OR381021 Razaghi et al. (2024)
Neopestalotiopsis egyptiaca CBS 140162T Egypt KP943747 KP943748 KP943746 Crous et al. (2015b)
Neopestalotiopsis elaeagni HGUP10002 T China MW930716 MZ203452 MZ683391 He et al. (2022)
Neopestalotiopsis elaeidis MFLUCC 15-0735T Thailand ON650690 ON734012 N/A Konta et al. (2023)
Neopestalotiopsis ellipsospora MFLUCC 12-0283T China JX398980 JX399047 JX399016 Maharachchikumbura et al. (2012)
Neopestalotiopsis eucalypticola CBS 264.37T N/A KM199376 KM199551 KM199431 Maharachchikumbura et al.(2014b)
Neopestalotiopsis eucalyptorum CBS 147684T Portugal MW794108 MW805397 MW802841 Diogo et al. (2021)
Neopestalotiopsis foedans CGMCC 3.9123T China JX398987 JX399053 JX399022 Maharachchikumbura et al. (2012)
Neopestalotiopsis formicarum CBS 362.72T Ghana KM199358 KM199517 KM199455 Maharachchikumbura et al.(2014b)
Neopestalotiopsis fragariae ZHKUCC 22- 0113T China ON553410 ON569076 ON569075 Prematunga et al. (2022)
Neopestalotiopsis guajavae FMBCC 11.1 Pakistan MF783085 MH460868 MH460871 Ul Haq et al. (2021)
Neopestalotiopsis guangxiensis CGMCC 3.23505T China OR247881 OR361440 OR381040 Razaghi et al. (2024)
Neopestalotiopsis guava UJFCC2084T Iran PP038121 PP053741 PP053735 This study
Neopestalotiopsis guava UJFCC2085 Iran PP038120 PP053740 PP053734 This study
Neopestalotiopsis guava UJFCC2086 Iran PP038122 PP053742 PP053736 This study
Neopestalotiopsis haikouensis SAUCC212271T China OK087294 OK104877 OK104870 Hsu et al. (2024)
Neopestalotiopsis hispanica CBS 147686T Portugal MW794107 MW805399 MW802840 Diogo et al. (2021)
Neopestalotiopsis honoluluana CBS 114495T USA KM199364 KM199548 KM199457 Maharachchikumbura et al.(2014b)
Neopestalotiopsis hydeana MFLUCC 20-0132T Thailand MW266069 MW251129 MW251119 Huanlauek et al. (2021)
Neopestalotiopsis iberica CBS 147688T Portugal MW794111 MW805402 MW802844 Diogo et al. (2021)
Neopestalotiopsis iranensis CBS 137768T Iran KM074048 KM074051 KM074057 Ayoubi and Soleimani (2016a)
Neopestalotiopsis javaensis CBS 257.31T Indonesia KM199357 KM199543 KM199437 Maharachchikumbura et al.(2014b)
Neopestalotiopsis keteleeriae MFLUCC 13-0915T China KJ023087 KJ023089 KJ023088 Song et al. (2014)
Neopestalotiopsis longiappendiculata CBS 147690T Portugal MW794112 MW805404 MW802845 Diogo et al. (2021)
Neopestalotiopsis macadamiae BRIP 63737cT Australia KX186604 KX186627 KX186654 Akinsanmi et al. (2017)
Neopestalotiopsis maddoxii BRIP 72266aT Australia MZ303782 MZ344167 MZ312675 Prasannath et al. (2021)
Neopestalotiopsis megabetaspora CGMCC 3.23474T China OR247875 OR361410 OR381010 Razaghi et al. (2024)
Neopestalotiopsis mesopotamica CBS 336.86T Turkey KM199362 KM199555 KM199441 Maharachchikumbura et al.(2014b)
Neopestalotiopsis mianyangensis CGMCC 3.23554T China OP546681 OP723490 OP672161 Li et al. (2022)
Neopestalotiopsis musae MFLUCC 15-0776T Thailand NR156311 KX789685 KX789686 Norphanphoun et al. (2019)
Neopestalotiopsis natalensis CBS 138.41T South Africa NR_156288 KM199552 KM199466 Maharachchikumbura et al. (2014a)
Neopestalotiopsis nebuloides BRIP 66617T Australia MK966338 MK977633 MK977632 Crous et al. (2020)
Neopestalotiopsis paeonia-suffruticosa CGMCC 3.23555T China OP082292 OP204794 OP235980 Li et al. (2022)
Neopestalotiopsis pernambucana URM 7148-01T Brazil KJ792466 KU306739 N/A Silvério et al. (2016)
Neopestalotiopsis perukae FMBCC 11.3T Pakistan MH209077 MH523647 MH460876 Ul Haq et al. (2021)
Neopestalotiopsis petila MFLUCC 17-1738T Thailand MK764276 MK764320 MK764342 Norphanphoun et al. (2019)
Neopestalotiopsis phangngaensis MFLUCC 18-0119T Thailand MH388354 MH388390 MH412721 Tibpromma et al. (2018)
Neopestalotiopsis photiniae MFLUCC 22-0129T China OP498008 OP753368 OP752131 Sun et al. (2023)
Neopestalotiopsis phyllostachydis CGMCC 3.23491T China OR247891 OR361423 OR381023 Razaghi et al. (2024)
Neopestalotiopsis piceana CBS 394.48T UK KM199368 KM199527 KM199453 Maharachchikumbura et al.(2014b)
Neopestalotiopsis psidii FMBCC 11.2T Pakistan MF783082 MH460874 MH477870 Ul Haq et al. (2021)
Neopestalotiopsis rhizophorae MFLUCC 17-1551T Thailand MK764277 MK764321 MK764343 Norphanphoun et al. (2019)
Neopestalotiopsis rhododendri GUCC 21504T China MW979577 MW980444 MW980443 Yang et al. (2021)
Neopestalotiopsis rosae CBS 101057T Zealand New KM199359 KM199523 KM199429 Maharachchikumbura et al.(2014b)
Neopestalotiopsis rosicola CFCC 51992T China KY885239 KY885243 KY885245 Norphanphoun et al. (2019)
Neopestalotiopsis samarangensis MFLUCC 12-0233T Thailand JQ968609 JQ968611 JQ968610 Maharachchikumbura et al.(2014b)
Neopestalotiopsis saprophytica MFLUCC 12-0282T China JX398982 JX399048 JX399017 Maharachchikumbura et al.(2014b)
Neopestalotiopsis scalabiensis CAA1029T Portugal MW969748 MW959100 MW934611 Santos et al. (2022)
Neopestalotiopsis siciliana AC46T Italy ON117813 ON107273 ON209162 Fiorenza et al. (2022)
Neopestalotiopsis sichuanensis CFCC 54338T China MW166231 MW199750 MW218524 Jiang et al. (2021)
Neopestalotiopsis sonneratiae MFLUCC 17-1745T Thailand MK764280 MK764324 MK764346 Norphanphoun et al. (2019)
Neopestalotiopsis steyaertii IMI 192475T Australia KF582796 KF582792 KF582794 Maharachchikumbura et al.(2014a)
Neopestalotiopsis subepidermalis CFCC 55160T China OK560699 OM622425 OM117690 Peng et al. (2022)
Neopestalotiopsis suphanburiensis MFLUCC 22-0126T Thailand OP497994 OP753372 OP752135 Sun et al. (2023)
Neopestalotiopsis surinamensis CBS 450.74T Suriname KM199351 KM199518 KM199465 Maharachchikumbura et al.(2014b)
Neopestalotiopsis terricola CGMCC 3.23553T China OP082294 OP204796 OP235982 Li et al. (2022)
Neopestalotiopsis thailandica MFLUCC 17-1730T Thailand MK764281 MK764325 MK764347 Norphanphoun et al. (2019)
Neopestalotiopsis vacciniicola CAA1055T Portugal MW969751 MW959103 MW934614 Santos et al. (2022)
Neopestalotiopsis umbrinospora MFLUCC 12-0285T China JX398984 JX399050 JX399019 Maharachchikumbura et al. (2012)
Neopestalotiopsis vaccinii CAA1059 T Portugal MW969747 MW959099 MW934610 Santos et al. (2022)
Neopestalotiopsis vheenae BRIP 72293aT Australia MZ303792 MZ344177 MZ312685 Prasannath et al. (2021)
Neopestalotiopsis vitis MFLUCC 15-1265T China KU140694 KU140676 KU140685 Jayawardena et al. (2016)
Neopestalotiopsis zakeelii BRIP 72282aT Australia MZ303789 MZ344174 MZ312682 Prasannath et al. (2021)
Neopestalotiopsis zimbabwana CBS 111495T Zimbabwe N/A KM199545 KM199456 Maharachchikumbura et al.(2014b)
Neopestalotiopsis sp. MEAN 1325 Portugal MW794102 MW805414 MW802835 Diogo et al. (2021)
Neopestalotiopsis sp. MEAN 1327 Portugal MW794105 MW805416 MW802838 Diogo et al. (2021)
Neopestalotiopsis sp. MEAN 1328 Spain MW794115 MW805417 MW802848 Diogo et al. (2021)
Neopestalotiopsis sp. PPS14 Peru MK860757 MN000341 MN000344 Rodríguez-Gálvez et al. (2022)
Neopestalotiopsis sp. PAK10 Peru MK860755 MN000339 MN000342 Rodríguez-Gálvez et al. (2022)
Neopestalotiopsis sp. PPS3 Peru MK860756 MN000340 MN000343 Rodríguez-Gálvez et al. (2022)
Neopestalotiopsis sp. CBS 664.94 Netherlands KM199354 KM199525 KM199449 Maharachchikumbura et al.(2014b)
Neopestalotiopsis sp. CBS 177.25 Unknown KM199370 KM199533 KM199445 Maharachchikumbura et al.(2014b)
Neopestalotiopsis sp. CFCC 54340 China MW166235 MW199754 MW218528 Jiang et al. (2021)
Neopestalotiopsis sp. ZX22B China MW166236 MW199755 MW218529 Jiang et al. (2021)
Pestalotiopsis colombiensis CBS 118553T Colombia KM199307 KM199488 KM199421 Maharachchikumbura et al.(2014b)
Pestalotiopsis diversiseta MFLUCC 12-0287T China NR_120187 JX399073 JX399040 Maharachchikumbura et al. (2012)
Robillarda Africana CBS 122.75T South Africa KR873253 MH554414 MH554656 Crous et al. (2015a); Liu et al. (2019)
Robillarda aquatic MFLUCC 21–0217T Thailand OL504777 N/A N/A Manawasinghe et al. (2022)
Robillarda australiana CBS 143882T Australia MH554091 MH554525 MH554764 Liu et al. (2019)
Robillarda khodaparastii UJFCC2116T Iran PP038123 PP053743 PP053737 This study
Robillarda khodaparastii UJFCC2117 Iran PP038124 PP053744 PP053738 This study
Robillarda khodaparastii UJFCC2118 Iran PP038125 PP053745 PP053739 This study
Robillarda mangiferae KUMCC 18-0180T China OL504777 N/A N/A Phookamsak et al. (2019)
Robillarda roystoneae CBS 115445T Hong Kong KR873254 KR873310 KR873317 Crous et al. (2015a)
Robillarda sessilis CBS 114312T Germany KR873256 KR873312 KR873319 Crous et al. (2015a)
Robillarda terrae CBS 587.71T India KJ710484 MH554493 MH554734 Crous et al. (2014); Liu et al. (2019)
Robillarda sp. CPC 25020 N/A KR873259 KR873315 KR873322 Crous et al. (2015a)

Maximum Likelihood (ML) analyses were performed using RAxML (Stamatakis 2006) as implemented in raxmlGUI 1.3 (Silvestro and Michalak 2012) with the ML + rapid bootstrap setting and the GTRGAMMA substitution model. A total of 1000 bootstrap replicates were conducted.

Maximum Parsimony (MP) analyses were performed with PAUP v. 4.0a169 (Swofford 2002). All molecular characters were treated as unordered and given equal weight, with gaps treated as missing data. The COLLAPSE command was set to MINBRLEN. MP analysis of the combined multilocus matrix was done using 1000 replicates of heuristic search with random addition of sequences, followed by TBR branch swapping (MULTREES option in effect, steepest descent option not in effect). Bootstrap analyses with 1000 replicates were performed similarly, with 10 rounds of random sequence addition and subsequent branch swapping during each bootstrap replicate. Bootstrap values ≤ 70% are considered low, those between 70% and 90% intermediate, and those ≥ 90% high.

Results

Molecular phylogeny

In Neopestalotiopsis, of the 1363 characters included in the phylogenetic analyses (ITS-tef1-tub2), 255 were parsimony-informative (48 in ITS, 120 in tef1, and 87 in tub2). The phylogram of the best ML tree (lnL = −6,428.7652) obtained using RAxML is shown in Fig. 1. The MP analysis revealed 896 trees with a length of 805 (not shown) that had a similar topology to the ML tree (CI = 0.62, RI = 0.71, and RC = 0.37).

Figure 1. 

Phylogram of the best ML trees (lnL = −6,428.7652) revealed by RAxML from an analysis of the combined ITS–tef1–tub2 matrix of selected Neopestalotiopsis. Strains in bold were sequenced in the current study. ML and MP bootstrap support above 50% are given at the first and second positions, respectively, above or below the branches.

The isolates of Neopestalotiopsis from this study form a clade with a well-supported ML and MP BS (80/70%). Table 2 shows the base pair differences among other taxa that might be mistaken for the new species.

Table 2
Download as
CSV
XLSX

Base pair differences between Neopestalotiopsis guava to related species in this study.

Species Gene region
ITS tef1 tub2
Neopestalotiopsis celtidis 4/434 10/432 5/371
Neopestalotiopsis eucalyptorum 1/505 10/475 1/340
Neopestalotiopsis foedans 5/505 6/475 0/340
Neopestalotiopsis hispanica 3/505 8/475 3/340
Neopestalotiopsis javaensis 2/505 3/475 0/340
Neopestalotiopsis maddoxii 6/505 7/475 1/340
Neopestalotiopsis megabetaspora 2/434 11/432 2/371
Neopestalotiopsis mesopotamica 2/505 9/475 1/340
Neopestalotiopsis perukae 5/505 1/475 1/340
Neopestalotiopsis rosae 2/505 7/475 0/340
Neopestalotiopsis scalabiensis 2/505 8/475 0/340
Neopestalotiopsis subepidermalis 3/456 8/432 3/371
Neopestalotiopsis suphanburiensis 2/434 8/411 2/371
Neopestalotiopsis vaccinii 2/505 8/475 3/340

In Robillarda, of the 1306 characters included in the phylogenetic analyses (ITS-tef1-tub2), 238 were parsimony-informative (48 in ITS, 144 in tef1, and 46 in tub2). The phylogram of the best ML tree (lnL = −3,732.5074) obtained using RAxML is presented in Fig. 2. The MP analysis revealed a single tree with a length of 417 (not shown) that exhibited a similar topology to the ML tree (CI = 0.91, RI = 0.91, and RC = 0.08).

Figure 2. 

Phylogram of the best ML trees (lnL = −3,732.5074) revealed by RAxML from an analysis of the combined ITS–tef1–tub2 matrix of selected Robillarda spp. Strains in bold were sequenced in the current study. ML and MP bootstrap support above 50% are given at the first and second positions, respectively, above or below the branches.

Robillarda isolates from the current study were grouped within a highly supported ML and MP bootstrap-supported clade, along with an unnamed isolate (CPC 25020). Analysis of these sequence data revealed identical sequences across all loci. This clade is a sister group of R. roystoneae (CBS 115445) with maximum ML and MP BS support. Molecularly, R. khodaparastii differs from R. roystoneae by 1 bp difference out of 532 bp in ITS, 2 bp differences out of 284 bp in tef1 and 2 bp differences out of 237 bp in tub2. Based on these findings, we conclude that members of Neopestalotiopsis and Robillarda represent two independent, so far undescribed species.

Taxonomy

Neopestalotiopsis guava A.R. Amirmijani, A. Pordel, & K. Dehghani, sp. nov.

MycoBank No: 85286
Fig. 3

Holotype.

Iran • Sistan and Baluchestan Province, Zar Abad region, from the infected leaves of Psidium guava, 5 November 2021, leg. A. Pordel and A.R. Amirmijani (holotype: CUJ0100; ex-type culture: UJFCC2084).

Etymology.

Named after the host plant, Psidium guava.

Description.

Conidiomata, solitary, black, and (300–)500–300(–700) m diam., and glistening conidial masses (Fig. 3E). Conidiogenous cells 6–10 × 2.5–4 µm, discrete, cylindrical, hyaline, smooth, thin-walled, simple, holoblastic-annelidic, percurrently, with collarette present. Conidia 18–30 × 5–7 µm, fusoid, four-septate, smooth, and slightly constricted at the septa; the basal cell thin walled, hyaline, 5–8 µm long; three median cells cylindrical, 15–18 µm long, smooth-walled, with septa darker than the rest of cell; the second cell from the base pale brown and 4–5 µm long; the third cell medium brown and 5 µm long; the fourth cell medium brown and 5 µm long; with septum between the third and fourth cell more distinct, broader, and darker brown than the other septa; the apical cell conic with a subacute apex, thin-walled, smooth, hyaline, 4–6 µm long, and with 2–3(–4) apical appendages (mostly 2) arising from the apical adage; apical appendages unbranched and straight, 11–24 µm long and up to 1 µm wide (n = 100); basal appendage single, filiform, unbranched, centric, 3–5 µm long, and up to 0.5 µm wide (n = 80).

Figure 3. 

Neopestalotiopsis guava (Holotype UJFCC2084) A, B surface and reverse of colony after 7 days on PDA C colonies on PNA after 7 days D colonies on CLA after 7 days E conidiomata with black sporulation on PNA F conidiomata with black sporulation on CLA G, H conidiogenous cells I, J conidia with apical and basal appendage. Scale bars: 2 mm (E, F); 20 µm (G, H); 10 µm (I, J).

Culture characteristics.

Colony on PDA attaining 70 mm diameter after 7 d at 25 °C, surface white and reverse pale cream (Fig. 3A, B), with fluffy white aerial mycelium, conidiomata scattered.

Other specimens examined.

Iran • Sistan and Baluchestan Province, Zar Abad region, from the infected leaves of Psidium guava, 5 November 2021, leg. A. Pordel and A.R. Amirmijani (cultures UJFCC2084 and UJFCC2086).

Notes.

Neopestalotiopsis guava is phylogenetically closely related to a large clade containing N. celtidis, N. ellipsospora, N. eucalyptorum, N. foedans, N. hispanica, N. javaensis, N. maddoxii, N. megabetaspora, N. mesopotamica, N. perukae, N. rosae, N. scalabiensis, N. subepidermalis, N. suphanburiensis and N. vaccinii. However, distinct morphological differences distinguish it from all these species (Table 3).

Table 3
Download as
CSV
XLSX

Morphological comparison of Neopestalotiopsis species related to this study.

Species Conidial size (µm) Apical appendages Basal appendage References
Number Length (µm) Length (µm)
N. celtidis 17.5–23.5 × 6–8 2–3(–4) (7.5–)11.5–21(–25) 2–6 Razaghi et al. (2024)
N. ellipsospora 19–25 × 5–6.5 1–3 5–12 3–4 Maharachchikumbura et al. (2012)
N. eucalyptorum (22.6)27.5–29.2(33.2) × (6.4)7.6–8.1(9.5) 3–4 (12.7)16.2–18.8(27.7) (3.4)5.4–6.2(8.1) Diogo et al. (2021)
N. foedans 19–24 × 5.7–6.9 2–3 6–18 3–6 Maharachchikumbura et al. (2012)
N. guava 18–30 × 5–7 2–3(–4) 11–24 × 1–1.2 3–5 × 0.4–0.5 This study
N. hispanica (21.4) 22.9–24.1 (29.4) × (7.2)8.2–8.7(9.8) 2–3 (13)18.2–20.3(24.6) (3.1)5.2–6.1(8.8) Diogo et al. (2021)
N. javaensis (24–)25–30(–31) × (6.5–)7–8.5(–9) 1–3 2–10(–18) 2–4 Maharachchikumbura et al. (2014b)
N. maddoxii 25–30 × 7–11 3 15–27 N/A Prasannath et al. (2021)
N. megabetaspora (19–)22–28 × 5.5–9 2–4 20–37 (2.5–)4.5–13 Razaghi et al. (2024)
N. mesopotamica (25–)26–32(–34) × (7–)7.5–9(–9.5) 3–4 (25–)28–38(–41) 4–6.5 Maharachchikumbura et al. (2014b)
N. perukae 19.7 ± 1.4 × 6.4 ± 0.8 2 22.2 ± 5.8 3.8 ± 1.9 Ul Haq et al. (2021)
N. rosae (20–)22–37(–29) × (7–)7.5–9.5(–10.5) 3–5 (22–)24–31(–33) 5–8 Maharachchikumbura et al. (2014b)
N. scalabiensis (10.3–)13.8–15.1(–23.3) × (3.7–)4.8–6.6(–5.3) 2–3 5.9–31.8 N/A Santos et al. (2022)
N. subepidermalis (19.5–)20–25(–26) × 7.5–9(–9.5) 2–4 (26.5–)27–32.5(–33.5) (6.5–)7–7.5(–8) Peng et al. (2022)
N. suphanburiensis 19–29 × 4–7 2–3 9–21 2–11 Sun et al. (2023)
N. vaccinii (11.0–)13.4–13.8(–15.2) × (4.9–)6.3–6.6(–7.5) 2–3 8.9–25.3 N/A Santos et al. (2022)

Robillarda khodaparastii A.R. Amirmijani, A. Pordel & K. Dehghani, sp. nov.

MycoBank No: 852862
Fig. 4

Holotype.

Iran • Sistan and Baluchestan Province, Ghasreghand Abad region, from the infected leaves of Mangifera indica, 15 December 2021, leg. A. Pordel and A.R. Amirmijani (holotype: CUJ0103; ex-type culture: UJFCC2116).

Etymology.

The species name is suggested as a tribute to our professor, Dr. Seyed Akbar Khodaparast, in recognition of his significant contributions to the progress of mycology in Iran.

Description.

Sexual morph undetermined. Asexual morph coelomycetous. Conidiomata 300–340 mm diam., black, semi-immersed, solitary, scattered, irregular shape, glabrous, minutely ostiolate. Conidiophores are reduced to conidiogenous cells. Conidiogenous cells 13–20 × 3–6 µm, holoblastic, proliferation percurrent 1–3 times, discrete, subcylindrical to ampulliform, hyaline, aseptate, smooth-walled. Conidia 12–13 × 2–3 µm (n = 50), hyaline, cylindrical, straight, 1-septate, thin and smooth-walled, apical cell developed into a branched appendage; appendages 12–20 × 1–2.5 µm (n = 50), dividing into 2 branches, straight, non- flexuous, broadly tubular, narrower towards the apex.

Culture characteristics.

Colony on PDA and MEA are similar, attaining 75 to 78 mm diameter after 7 d at 25 °C, surface and reverse white to cream (Fig. 4A, B), with fluffy white aerial mycelium, conidiomata scattered.

Figure 4. 

Robillarda khodaparastii (Holotype UJFCC2116) A, B surface and reverse of colony after 7 days on PDA C colonies on PNA after 7 days D colonies on CLA after 7 days E conidiomata with brown sporulation on CLA F, G conidiogenous cells H–L conidia with apical appendage. Scale bars: 2 mm (E); 10 µm (F–L).

Other specimen examined.

Iran • Sistan and Baluchestan Province, Ghasreghand Abad region, from the infected leaves of Mangifera indica, 15 December 2021, leg. A. Pordel and A.R. Amirmijani (cultures UJFCC2117 and UJFCC2118).

Notes.

A comparison of sequence data revealed complete identities among the Robillarda isolates. This clade was determined as a sister group of R. roystoneae (CBS 115445) with maximum ML and MP bootstrap support. R. khodaparastii is morphologically and phylogenetically near to R. roystoneae, but our species can be easily distinguished from the latter species by producing longer conidiogenous cells [13–20 × 3–6 vs. 7–12 × 2–3 µm] and shorter conidia [12–13 × 2–3 vs. (13–)14–15(–16) × 2.5–3(–3.5) µm].

Discussion

Mango (Mangifera indica L.) and Guava (Psidium guava L.) are popular fruits in tropical and subtropical regions due to their delicious taste, high nutritional value, and economic importance in international markets (Kumar et al. 2021). The cultivation of mango has expanded beyond traditional and non-traditional production countries like the United Arab Emirates (UAE) (Saeed et al. 2017). Iran’s diverse climates, characterized by significant temperature variations between the northern and southern regions, allow for the production of various agricultural products (Saboki et al. 2012).

Several pestalotioid fungi have been reported from diverse hosts in Iran, including: Pestalotia disseminata, Pestalotiopsis acacia, P. biciliata, P. funereal, P. longiseta, P. longisetula, P. neglecta, P. nattrassii, P. trachycarpicola, P. vismiae, P. uvicola, Pseudopestalotiopsis theae, and Neopestalotiopsis asiatica, N. iranensis, N. mesopotamica (Khodaparast and Hedjaroude 1994; Borhani and Mousazadeh 2004; Arzanlou et al. 2012; Naeimi et al. 2015; Ayoubi and Soleimani 2016a, 2016b; Atashi Khalilabad and Fotouhifar 2022; Bakhshi et al. 2022; Amirmijani et al. 2024).

According to Maharachchikumbura et al. (2011, 2013), many species of Pestalotioid fungi have been named based on their host associations due to the lack of reliable distinctive features. However, recent research has demonstrated that many introduced phylogenetic species within Pestalotioid fungi can be distinguished using combined sequence data from ITS-rDNA, β-tubulin (tub2), and tef-1α genes. In this study, we employed these gene sequences, along with morphological features, for phylogenetic analysis and species identification. As a result, we described two new species of pestalotioid fungi from Iran. These novel species contribute to a deeper understanding of the taxonomy and diversity of Neopestalotiopsis and Robillarda in Iran. However, it is likely that our findings represent only the tip of the iceberg.

These fungi were found on leaf spots of mango and guava in southern Iran. Generally, these genera are regarded as insignificant pathogens, however, they have been observed to cause diseases in various crops (Maharachchikumbura et al. 2012). These fungi often act as endophytes or saprobes, and their pathogenic roles still remain little studied and, therefore, inadequately understood. Although these species were isolated from leaf spots, we were unable to conduct pathogenicity tests in this study. Consequently, further research is necessary to evaluate their potential aggressiveness and negative effects on tropical fruits. Accurate species identification of plant-pathogenic genera (Jayawardena et al. 2015), such as Pestalotiopsis-like fungi, is crucial for plant pathologists in managing and controlling plant diseases. Ongoing studies aim to clarify the environmental factors contributing to leaf spot disease to develop effective control measures.

Acknowledgements

The authors thank the Research Deputy of the University of Jiroft, Kerman, Iran, and the Plant Protection Research Department, Baluchestan Agricultural and Natural Resources Research and Education Center, AREEO, Iranshahr, Iran for financial support. We also extend our gratitude to Prof. Sajeewa S. N. Maharachchikumbura, Prof. Ning Jiang, and the other reviewers for reviewing the manuscript and providing insightful suggestions for its improvement.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This work was supported by the Research Deputy of the University of Jiroft, Kerman, Iran, and the Plant Protection Research Department, Baluchestan Agricultural and Natural Resources Research and Education Center, AREEO, Iranshahr, Iran. The study was also supported by the Biodiversa+ Project FUNACTION of the German Science foundation (GR1540/47-1) provided to HPG.

Author contributions

AA, AP: conceptualization, isolation and identification of the producer strain, writing - original draft preparation, review and editing; KS: isolation of the strains; MJP: phylogenetic analysis, writing - review and editing; HM, H-PG: revision and funding.

Author ORCIDs

Amirreza Amirmijani https://orcid.org/0000-0002-4529-4415

Adel Pordel https://orcid.org/0000-0001-5999-9443

Kowsar Dehghani https://orcid.org/0009-0004-2930-3491

Mohammad Javad Pourmoghaddam https://orcid.org/0000-0001-6055-7503

Hans-Peter Grossart https://orcid.org/0000-0002-9141-0325

Data availability

The data that support the findings of this study are available in GenBank (https://www.ncbi.nlm.nih.gov/genbank/) and in culture collections and fungal herbarium, as shown in Table 1 and the text.

References

  • Akinsanmi OA, Nisa S, Jeff-Ego OS, Shivas RG, Drenth A (2017) Dry flower disease of macadamia in australia caused by Neopestalotiopsis macadamiae sp. nov. and Pestalotiopsis macadamiae sp. nov. Plant Disease 101(1): 45–53. https://doi.org/10.1094/PDIS-05-16-0630-RE
  • Atashi Khalilabad A, Fotouhifar K (2022) Morphological and molecular characterization of a novel Pestalotiopsis trachycarpicola, causing Garden Croton leaf spot in Iran. Mycologia Iranica 9: 59–66. https://doi.org/10.22043/MI.2022.360270.1236
  • Ayoubi N, Soleimani MJ (2016b) Morphological and molecular identification of Neopestalotiopsis asiatica causing leaf spot on sweet almond. Journal of Plant Pathology 98(2): 321–325. https://doi.org/10.4454/JPP.V98I2.030
  • Bezerra JDP, Machado AR, Firmino AL, Rosado AWC, Souza CAF, Souza-Motta CM, Freire KTLS, Paiva LM, Magalhães OMC, Pereira OL, Crous PW, Oliveira TGL, Abreu VP, Fan X (2018) Mycological diversity description I. Acta Botanica Brasílica 32(4): 656–666. https://doi.org/10.1590/0102-33062018abb0154
  • Borhani A, Mousazadeh S (2004) Study on the hosts and pathogenicity of Pestalotiopsis funerea (Desm.) Steyaert on some soft wood species in Mazandaran Province. Proceedings of the 16th Iranian Plant Protection Congress, Vol. II, 28 Aug.–1 Sept. , Tabriz, Iran, 441 pp.
  • Crous PW, Giraldo A, Hawksworth DL, Robert V, Kirk PM, Guarro J, Robbertse B, Schoch CL, Damm U, Trakunyingcharoen T, Groenewald JZ (2014) The Genera of Fungi: Fixing the application of type species of generic names. IMA Fungus 5(1): 141–160. https://doi.org/10.5598/imafungus.2014.05.01.14
  • Crous PW, Carris LM, Giraldo A, Groenewald JZ, Hawksworth DL, Hemández-Restrepo M, Jaklitsch WM, Lebrun M-H, Schumacher RK, Stielow JB, van der Linde EJ, Vilcāne J, Voglmayr H, Wood AR (2015a) The genera of fungi-fixing the application of the type species of generic names–G 2: Allantophomopsis, Latorua, Macrodiplodiopsis, Macrohilum, Milospium, Protostegia, Pyricularia, Robillarda, Rotula, Septoriella, Torula, and Wojnowicia. IMA Fungus 6(1): 163–198. https://doi.org/10.5598/imafungus.2015.06.01.11
  • Crous PW, Wingfield MJ, Roux JJ le, Richardson DM, Strasberg D, Shivas RG, Alvarado P, Edwards J, Moreno G, Sharma R, Sonawane MS, Tan YP, Altés A, Barasubiye T, Barnes CW, Blanchette RA, Boertmann D, Bogo A, Carlavilla JR, Cheewangkoon R, Daniel R, de Beer ZW, Yáñez-Morales M de Jesús, Duong TA, Fernández-Vicente J, Geering ADW, Guest DI, Held BW, Heykoop M, Hubka V, Ismail AM, Kajale SC, Khemmuk W, Kolařík M, Kurli R, Lebeuf R, Lévesque CA, Lombard L, Magista D, Manjón JL, Marincowitz S, Mohedano JM, Nováková A, Oberlies NH, Otto EC, Paguigan ND, Pascoe IG, Pérez-Butrón JL, Perrone G, Rahi P, Raja HA, Rintoul T, Sanhueza RMV, Scarlett K, Shouche YS, Shuttleworth LA, Taylor PWJ, Thorn RG, Vawdrey LL, Solano-Vidal R, Voitk A, Wong PTW, Wood AR, Zamora JC, Groenewald JZ (2015b) Fungal planet description sheets: 371–399. Persoonia 35(1): 264–327. https://doi.org/10.3767/003158515X690269
  • Crous PW, Wingfield MJ, Chooi YH, Gilchrist CLM, Lacey E, Pitt JI, Roets F, Swart WJ, Cano-Lira JF, Valenzuela-Lopez N, Hubka V, Shivas RG, Stchigel AM, Holdom DG, Jurjević Ž, Kachalkin AV, Lebel T, Lock C, Martín MP, Tan YP, Tomashevskaya MA, Vitelli JS, Baseia IG, Bhatt VK, Brandrud TE, De Souza JT, Dima B, Lacey HJ, Lombard L, Johnston PR, Morte A, Papp V, Rodríguez A, Rodríguez-Andrade E, Semwal KC, Tegart L, Abad ZG, Akulov A, Alvarado P, Alves A, Andrade JP, Arenas F, Asenjo C, Ballarà J, Barrett MD, Berná LM, Berraf-Tebbal A, Bianchinotti MV, Bransgrove K, Burgess TI, Carmo FS, Chávez R, Čmoková A, Dearnaley JDW, Santiago ALCM de A, Freitas-Neto JF, Denman S, Douglas B, Dovana F, Eichmeier A, Esteve-Raventós F, Farid A, Fedosova AG, Ferisin G, Ferreira RJ, Ferrer A, Figueiredo CN, Figueiredo YF, Reinoso-Fuentealba CG, Garrido-Benavent I, Cañete-Gibas CF, Gil-Durán C, Glushakova AM, Gonçalves MFM, González M, Gorczak M, Gorton C, Guard FE, Guarnizo AL, Guarro J, Gutiérrez M, Hamal P, Hien LT, Hocking AD, Houbraken J, Hunter GC, Inácio CA, Jourdan M, Kapitonov VI, Kelly L, Khanh TN, Kisło K, Kiss L, Kiyashko A, Kolařík M, Kruse J, Kubátová A, Kučera V, Kučerová I, Kušan I, Lee HB, Levicán G, Lewis A, Liem NV, Liimatainen K, Lim HJ, Lyons MN, Maciá-Vicente JG, Magaña-Dueñas V, Mahiques R, Malysheva EF, Marbach PAS, Marinho P, Matočec N, McTaggart AR, Mešić A, Morin L, Muñoz-Mohedano JM, Navarro-Ródenas A, Nicolli CP, Oliveira RL, Otsing E, Ovrebo CL, Pankratov TA, Paños A, Paz-Conde A, Pérez-Sierra A, Phosri C, Pintos Á, Pošta A, Prencipe S, Rubio E, Saitta A, Sales LS, Sanhueza L, Shuttleworth LA, Smith J, Smith ME, Spadaro D, Spetik M, Sochor M, Sochorová Z, Sousa JO, Suwannasai N, Tedersoo L, Thanh HM, Thao LD, Tkalčec Z, Vaghefi N, Venzhik AS, Verbeken A, Vizzini A, Voyron S, Wainhouse M, Whalley AJS, Wrzosek M, Zapata M, Zeil-Rolfe I, Groenewald JZ (2020) Fungal planet description sheets: 1042–1111. Persoonia - Molecular Phylogeny and Evolution Fungi 44: 301–459. https://doi.org/10.3767/persoonia.2020.44.11
  • Diogo E, Gonçalves CI, Silva AC, Valente C, Bragança H, Phillips AJ (2021) Five new species of Neopestalotiopsis associated with diseased Eucalyptus spp. in Portugal. Mycological Progress 20(11): 1441–1456. https://doi.org/10.1007/s11557-021-01741-5
  • FAO (2020) Major Tropical Fruits Market Review 2019. Rome.
  • FAO (2024) Major Tropical Fruits Market Review. Preliminary Results 2023. Rome.
  • Fiorenza A, Gusella G, Aiello D, Polizzi G, Voglmayr H (2022) Neopestalotiopsis siciliana sp. nov. and N. rosae causing stem lesion and dieback on avocado plants in Italy. Journal of Fungi 8(6): 562. https://doi.org/10.3390/jof8060562
  • Glass NL, Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61(4): 1323–1330. https://doi.org/10.1128/aem.61.4.1323-1330.1995
  • He YK, Yang Q, Sun YR, Zeng XY, Jayawardena RS, Hyde KD, Wang Y (2022) Additions to Neopestalotiopsis (Amphisphaeriales, Sporocadaceae) fungi: Two new species and one new host record from China. Biodiversity Data Journal 10: e90709. https://doi.org/10.3897/BDJ.10.e90709
  • Hsu SY, Xu YC, Lin YC, Chuang WY, Lin SR, Stadler M, Tangthirasunun N, Cheewangkoon R, AL-Shwaiman HA, Elgorban AM, Ariyawansa HA (2024) Hidden diversity of Pestalotiopsis and Neopestalotiopsis (Amphisphaeriales, Sporocadaceae) species allied with the stromata of entomopathogenic fungi in Taiwan. MycoKeys 101: 275–312. https://doi.org/10.3897/mycokeys.101.113090
  • Huanlauek N, Jayawardena RS, Maharachchikumbura SSN, Harishchandra DL (2021) Additions to pestalotioid fungi in Thailand: Neopestalotiopsis hydeana sp. nov., Pestalotiopsis hydei sp. nov. Phytotaxa 479(1): 23–43. https://doi.org/10.11646/phytotaxa.479.1.2
  • Jayawardena RS, Zhang W, Liu M, Maharachchikumbura SSN, Zhou Y, Huang JB, Nilthong S, Wang ZY, Li XH, Yan JY, Hyde KD (2015) Identification and characterization of Pestalotiopsis-like fungi related to grapevine diseases in China. Fungal Biology 119(5): 348–361. https://doi.org/10.1016/j.funbio.2014.11.001
  • Jayawardena RS, Liu M, Maharachchikumbura SSN, Zhang W, Xing QK, Hyde KD, Nilthong S, Li XH, Yan JY (2016) Neopestalotiopsis vitis sp. nov. causing grapevine leaf spot in China. Phytotaxa 258(1): 63–74. https://doi.org/10.11646/phytotaxa.258.1.4
  • Jiang N, Fan X, Tian C (2021) Identification and characterization of leaf-inhabiting fungi from Castanea plantations in China. Journal of Fungi 7(1): 64. https://doi.org/10.3390/jof7010064
  • Jiang N, Voglmayr H, Xue H, Piao CG, Li Y (2022) Morphology and Phylogeny of Pestalotiopsis (Sporocadaceae, Amphisphaeriales) from Fagaceae Leaves in China. Microbiology Spectrum 10(6): 1–28. https://doi.org/10.1128/spectrum.03272-22
  • Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: Multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20(4): 1160–1166. https://doi.org/10.1093/bib/bbx108
  • Khodaparast SA, Hedjaroude GhA (1994) Incidence of a new Pestalotiopsis species in association with grey blight of tea in Guilan. Iranian Journal of Plant Pathology 30: 79–80 [in Persian], 33 [in English].
  • Konta S, Tibpromma S, Karunarathna SC, Samarakoon MC, Steven LS, Mapook A, Boonmee S, Senwanna C, Balasuriya A, Eungwanichayapant PD, Hyde KD (2023) Morphology and multigene phylogeny reveal ten novel taxa in Ascomycota from terrestrial palm substrates (Arecaceae) in Thailand. Mycosphere 14(1): 107–152. https://doi.org/10.5943/mycosphere/14/1/2
  • Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7): 1870–1874. https://doi.org/10.1093/molbev/msw054
  • Kumar V, Cheewangkoon R, Gentekaki E, Maharachchikumbura SSN, Brahmanage RS, Hyde KD (2019) Neopestalotiopsis alpapicalis sp. nov. a new endophyte from tropical mangrove trees in Krabi Province (Thailand). Phytotaxa 393(3): 251–262. https://doi.org/10.11646/phytotaxa.393.3.2
  • Kumar M, Tomar M, Amarowicz R, Saurabh V, Nair MS, Maheshwari C, Sasi M, Prajapati U, Hasan M, Singh S, Changan S, Prajapat RK, Berwal MK, Satankar V (2021) Guava (Psidium guajava L.) leaves: Nutritional composition, phytochemical profile, and health-promoting bioactivities. Foods 10(4): 752. https://doi.org/10.3390/foods10040752
  • Li WL, Dissanayake AJ, Zhang T, Maharachchikumbura SSN, Liu JK (2022) Identification and pathogenicity of Pestalotiod Fungi associated with woody oil plants in Sichuan Province, China. Journal of Fungi 8(11): 1175. https://doi.org/10.3390/jof8111175
  • Liu F, Bonthond G, Groenewald JZ, Cai L, Crous PW (2019) Sporocadaceae, a family of coelomycetous fungi with appendage-bearing conidia. Studies in Mycology 92(1): 287–415. https://doi.org/10.1016/j.simyco.2018.11.001
  • Liu X, Tibpromma S, Zhang F, Xu J, Chethana KWT, Karunarathna SC, Mortimer PE (2021) Neopestalotiopsis cavernicola sp. nov. from Gem Cave in Yunnan Province, China. Phytotaxa 512(1): 1–27. https://doi.org/10.11646/phytotaxa.512.1.1
  • Ma XY, Maharachchikumbura SSN, Chen BW, Hyde KD, McKenzie EHC, Chomnunti P, Kang J-C (2019) Endophytic pestalotiod taxa in Dendrobium orchids. Phytotaxa 419(3): 268–286. https://doi.org/10.11646/phytotaxa.419.3.2
  • Maharachchikumbura SSN, Guo LD, Chukeatirote E, Bahkali AH, Hyde KD (2011) Pestalotiopsis—Morphology, phylogeny, biochemistry and diversity. Fungal Diversity 50(1): 167–187. https://doi.org/10.1007/s13225-011-0125-x
  • Maharachchikumbura SSN, Guo LD, Cai L, Chukeatirote E, Wu WP, Sun X, Crous PW, Bhat DJ, McKenzie EHC, Bahkali AH, Hyde KD (2012) A multilocus backbone tree for Pestalotiopsis, with a polyphasic characterization of 14 new species. Fungal Diversity 56(1): 95–129. https://doi.org/10.1007/s13225-012-0198-1
  • Maharachchikumbura SSN, Guo LD, Chukeatirote E, McKenzie EHC, Hyde KD (2013) A destructive new disease of Syzygium samarangense in Thailand caused by the new species Pestalotiopsis samarangensis. Tropical Plant Pathology 38(3): 227–235. https://doi.org/10.1590/S1982-56762013005000002
  • Maharachchikumbura SSN, Guo LD, Chukeatirote E, Hyde KD (2014a) Improving the backbone tree for the genus Pestalotiopsis; addition of P. steyaertii and P. magna sp. nov. Mycological Progress 13(3): 617–624. https://doi.org/10.1007/s11557-013-0944-0
  • Maharachchikumbura SSN, Larignon P, Hyde KD, Al-Sadi AM, Liu ZY (2016) Characterization of Neopestalotiopsis, Pestalotiopsis and Truncatella species associated with grapevine trunk diseases in France. Phytopathologia Mediterranea 55(3): 380–390. https://doi.org/10.14601/Phytopathol_Mediterr-18298
  • Manawasinghe IS, Calabon MS, Jones EBG, Zhang YX, Liao CF, Xiong YR, Chaiwan N, Kularathnage ND, Liu NG, Tang SM, Sysouphanthong P, Du TY, Luo M, Pasouvang P, Pem D, Phonemany M, Ishaq M, Chen JW, Karunarathna SC, Mai ZL, Rathnayaka AR, Samarakoon MC, Tennakoon DS, Wijesinghe SN, Yang YH, Zhao HJ, Fiaz M, Doilom M, Dutta AK, Khalid AN, Liu JW, Thongklang N, Senanayake IC, Tibpromma S, You LQ, Camporesi E, Gafforov YS, Hyde KD KD (2022) Mycosphere notes 345–386. Mycosphere 13(1): 454–557. https://doi.org/10.5943/mycosphere/13/1/3
  • Marin-Felix Y, Hernández-Restrepo M, Iturrieta-González I, García D, Gené J, Groenewald JZ, Cai L, Chen Q, Quaedvlieg W, Schumacher RK, Taylor PWJ, Ambers C, Bonthond G, Edwards J, Krueger-Hadfield SA, Luangsa-Ard JJ, Morton L, Moslemi A, Sandoval-Denis M, Tan YP, Thangavel R, Vaghefi N, Cheewangkoon R, Crous PW (2019) Genera of phytopathogenic fungi: GOPHY 3. Studies in Mycology 94: 1–124. https://doi.org/10.1016/j.simyco.2019.05.001
  • Naeimi S, Javadi L, Javadi AR (2015) First report of Pestalotia disseminata, the causal agent of feijoa fruit rot in Iran. Mycologia Iranica 2(1): 75–76. https://doi.org/10.22043/mi.2015.14228
  • Norphanphoun C, Jayawardena RS, Chen Y et al. (2019) Morphological and phylogenetic characterization of novel pestalotioid species associated with mangroves in Thailand. Mycosphere 10(1): 531–578. https://doi.org/10.5943/mycosphere/10/1/9
  • O’Donnell K, Kistler HC, Cigelnik E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing Panama disease of banana: Concordant evidence from nuclear and mitochondrial gene genealogies. Proceedings of the National Academy of Sciences of the United States of America 95(5): 2044–2049. https://doi.org/10.1073/pnas.95.5.2044
  • Phookamsak R, Hyde KD, Jeewon R, Bhat DJ, Jones EBG, Maharachchikumbura SSN, Raspé O, Karunarathna SC, Wanasinghe DN, Hongsanan S, Doilom M, Tennakoon DS, Machado AR, Firmino AL, Ghosh A, Karunarathna A, Mešić A, Dutta AK, Thongbai B, Devadatha B, Norphanphoun C, Senwanna C, Wei D, Pem D, Ackah FK, Wang G-N, Jiang H-B, Madrid H, Lee HB, Goonasekara ID, Manawasinghe IS, Kušan I, Cano J, Gené J, Li J, Das K, Acharya K, Raj KNA, Latha KPD, Chethana KWT, He M-Q, Dueñas M, Jadan M, Martín MP, Samarakoon MC, Dayarathne MC, Raza M, Park MS, Telleria MT, Chaiwan N, Matočec N, de Silva NI, Pereira OL, Singh PN, Manimohan P, Uniyal P, Shang Q-J, Bhatt RP, Perera RH, Alvarenga RLM, Nogal-Prata S, Singh SK, Vadthanarat S, Oh S-Y, Huang S-K, Rana S, Konta S, Paloi S, Jayasiri SC, Jeon SJ, Mehmood T, Gibertoni TB, Nguyen TTT, Singh U, Thiyagaraja V, Sarma VV, Dong W, Yu X-D, Lu Y-Z, Lim YW, Chen Y, Tkalčec Z, Zhang Z-F, Luo Z-L, Daranagama DA, Thambugala KM, Tibpromma S, Camporesi E, Bulgakov TS, Dissanayake AJ, Senanayake IC, Dai DQ, Tang L-Z, Khan S, Zhang H, Promputtha I, Cai L, Chomnunti P, Zhao R-L, Lumyong S, Boonmee S, Wen T-C, Mortimer PE, Xu J (2019) Fungal diversity notes 929–1035: Taxonomic and phylogenetic contributions on genera and species of fungi. Fungal Diversity 95(1): 1–273. https://doi.org/10.1007/s13225-019-00421-w
  • Prasannath K, Shivas RG, Galea VJ, Akinsanmi OA (2021) Neopestalotiopsis species associated with flower diseases of Macadamia integrifolia in Australia. Journal of Fungi 7(9): 771. https://doi.org/10.3390/jof7090771
  • Prematunga CJ, You LQ, Gomdola D, Balasuriya A, Yang YH, Jayawardena RS, Luo M (2022) An addition to pestalotioid fungi in China: Neopestalotiopsis fragariae sp. nov. causing leaf spots on Fragaria ananassa. Asian Journal of Mycology 5(2): 220–238. https://doi.org/10.5943/ajom/5/2/10
  • Refaei J, Jones EBG, Sakayaroj J, Santhanam J (2011) Endophytic fungi from Rafflesia cantleyi: Species diversity and antimicrobial activity. Mycosphere 2: 429–447.
  • Rodríguez-Gálvez E, Hilário S, Lopes A, Alves A (2022) Diversity and pathogenicity of Lasiodiplodia and Neopestalotiopsis species associated with stem blight and dieback of blueberry plants in Peru. European Journal of Plant Pathology 157(1): 89–102. https://doi.org/10.1007/s10658-020-01983-1
  • Saboki E, Usha K, Singh B (2012) Plant architectural traits and their role in defense mechanism against malformation in mango (Mangifera indica L.). Scientia Horticulturae 139: 25–31. https://doi.org/10.1016/j.scienta.2012.02.025
  • Saboki E, Usha K, Singh B (2014) Screening for resistance to malformation in mango (Mangifera indica L.). Progressive Horticulture 46: 209–216.
  • Saccardo PA (1880) Conspectus generum fungorum italiae inferiorum nempe ad Sphaeropsideas, Melanconieas et Hyphomyceteas pertinentium, systemate sporologico dispositu. Michelia 2: 1–38.
  • Saeed EE, Sham A, AbuZarqa A, Al Shurafa K, Al Naqbi T, Iratni R, El-Tarabily K, AbuQamar S (2017) Detection and Management of Mango Dieback Disease in the United Arab Emirates. International Journal of Molecular Sciences 18(10): 2086. https://doi.org/10.3390/ijms18102086
  • Santos J, Hilário S, Pinto G, Alves A (2022) Diversity and pathogenicity of pestalotioid fungi associated with blueberry plants in Portugal, with description of three novel species of Neopestalotiopsis. European Journal of Plant Pathology 162(3): 539–555. https://doi.org/10.1007/s10658-021-02419-0
  • Shu J, Yu Z, Sun W, Zhao J, Li Q, Tang L, Guo T, Huang S, Mo J, Hsiang T, Luo S (2020) Identification and characterization of pestalotioid fungi causing leaf spots on mango in southern China. Plant Disease 104(4): 1207–1213. https://doi.org/10.1094/PDIS-03-19-0438-RE
  • Song Y, Maharachchikumbura SS, Jiang YL, Hyde KD, Wang Y (2014) Pestalotiopsis keteleeria sp. nov., isolated from Keteleeria pubescens in China. Warasan Khana Witthayasat Maha Witthayalai Chiang Mai 41: 885–893.
  • Sun YR, Jayawardena RS, Sun JE, Wang Y (2023) Pestalotioid species associated with medicinal plants in southwest China and Thailand. Microbiology Spectrum 11(1): e03987–22. https://doi.org/10.1128/spectrum.03987-22
  • Sutton BC (1980) The Coelomycetes: Fungi Imperfecti with Pycnidia Acervuli and Stromata. Commonwealth Mycological Institute, Kew, 696 pp.
  • Swofford DL (2002) PAUP* 4.0b10: phylogenetic analysis using parsimony (*and other methods). Sinauer Associates, Sunderland.
  • Tibpromma S, Hyde KD, McKenzie EHC, Bhat DJ, Phillips AJL, Wanasinghe DN, Samarakoon MC, Jayawardena RS, Dissanayake AJ, Tennakoon DS, Doilom M, Phookamsak R, Tang AMC, Xu J, Mortimer PE, Promputtha I, Maharachchikumbura SSN, Khan S, Karunarathna SC (2018) Fungal diversity notes 840–928: Micro-fungi associated with Pandanaceae. Fungal Diversity 93(1): 1–160. https://doi.org/10.1007/s13225-018-0408-6
  • Ul Haq I, Ijaz S, Khan NA (2021) Genealogical concordance of phylogenetic species recognition-based delimitation of Neopestalotiopsis species associated with leaf spots and fruit canker disease affected guava plants. Pakistan Journal of Agricultural Sciences 58(4): 1301–1313. https://doi.org/10.21162/PAKJAS/21.1045
  • White TJ, Bruns TD, Lee S, Taylor S (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (Eds) PCR - Protocols and Applications - A Laboratory Manual, Academic Press, New York, 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
  • Yang Q, Zeng XY, Yuan J, Zhang Q, He YK, Wang Y (2021) Two new species of Neopestalotiopsis from southern China. Biodiversity Data Journal 9: e70446. https://doi.org/10.3897/BDJ.9.e70446

Amirreza Amirmijani and Adel Pordel contributed equally to this work.

Supplementary material

Supplementary material 1 

Phylogenetic trees

Amirreza Amirmijani, Adel Pordel, Kowsar Dehghani, Mohammad Javad Pourmoghaddam, Hossein Masigol, Hans-Peter Grossart

Data type: rar

Explanation note: Single gene trees for all loci for both genera.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (2.74 MB)
login to comment