Pseudoplagiostoma perseae sp. nov. causes leaf spot disease on avocado leaves in Taiwan

Pseudoplagiostoma perseae sp. nov. causes leaf spot disease on avocado leaves in Taiwan | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Pseudoplagiostoma perseae sp. nov. causes leaf spot disease on avocado leaves in Taiwan Chao-jung Wu, Jin Liang Chen, Shean Shong Tzean, Hui Fang Ni This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4019244/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 Aug, 2024 Read the published version in European Journal of Plant Pathology → Version 1 posted 6 You are reading this latest preprint version Abstract Leaf spot disease is commonly found on leaves of avocado plants in avocado orchards in Taiwan. Needle-like spots with yellow halos appear on diseased avocado leaves at the incipient stage, and these spots gradually enlarge and become brown lesions with black-brown centers. The causal agent of this disease is unknown. A Pseudoplagiostoma species was collected from these leaf spots. Evaluation of morphological traits and phylogenetic analysis using the ribosomal DNA internal transcribed spacer operon, β-tubulin, and partial large subunit of ribosomal DNA markers identified this fungal species as a novel species, Pseudoplagiostoma perseae sp. nov. A pathogenicity test was conducted on three avocado cultivars, ‘Choquette,’ ‘Hall,’ and ‘Hung Shin Yuan.’ After 2 weeks of inoculation, needle-like leaf spots appeared on all three cultivars, and P. perseae was re-isolated from the spots of all inoculated leaves, fulfilling Koch’s postulates. This is the first report describing the novel species P. perseae , which was validated as the causal agent of avocado leaf spot disease. Pseudoplagiostoma perseae avocado leaf spot disease pathogenicity test Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Avocado is one of the most popular tropical fruit crops in the world. According to the Food and Agriculture Organization (FAO) of the United Nations, 2.4 million tons of avocado were exported globally in 2022 (FAO 2023 ). In Taiwan, the avocado planting area was about 1,811 hectares in 2022, which represents a 147% increase from the planting area in 2016 (733 hectares). Some cultivars, such as ‘Changan,’ Choquette,’ ‘Hall,’ and ‘Hung Shin Yuan’ are commonly planted in orchards. There are three main pathogens known to cause lesions on avocado leaves. Cercospora purpurea (Pohronezny et al., 1994 ) causes Cercospora spot or blotch disease, which is initially characterized by small light-brown spots appearing on leaves, which later turn into angular brown lesions with a small yellow halo. Sometimes greyish fruiting bodies are visible in lesion centers under high humidity (Ruehle 1943 ; Stevens 1992 ; Zentmyer 1953 ). Colletotrichum species are also pathogenic on avocado leaves and cause symptoms including brown spots randomly spread on the tips of senescing leaves (Giblin et al., 2010 ; Pohronezny et al., 1994 ). Sphaceloma perseae causes avocado scab on fruits as well as leaves (Jenkins 1925 , 1934 ). The lesions on avocado leaves are grayish, oval to elongated in shape, and slightly elevated; they are mainly found on the reverse side of the leaf and mostly confined to the midrib and veins (Pegg et al., 2002 ). So far, no avocado leaf disease caused by Pseudoplagiostoma species has been reported. Currently, the genus Pseudoplagiostoma contains 15 species (Bezerra et al., 2019 ; Cheewangkoon et al., 2010 ; Crous et al., 2012 , 2018 ; Gomdola et al., 2023 ; Hittanadurage et al., 2023; Mu et al., 2022 ; Phookamsak et al., 2019 ; Suwannarach et al., 2016 ; Tang et al., 2022 ; Zhang et al., 2023 ). The asexual structures of the genus Pseudoplagiostoma include acervular to pycnidial conidiomata with cylindrical to ampulliform conidiogenous cells and aseptate ellipsoidal conidia; the sexual structures are depressed globose or elliptical ostiolate perithecia with subcylindrical to long obovoid asci, which bear eight, two seriate, hyaline, medianly one-septate ellipsoidal ascospores with tapering ends and terminal elongate appendages (Cheewangkoon et al., 2010 ). Pseudoplagiostoma species are distinguished using morphological characteristics of the conidia (size, color, and shape) (Suwannarach et al., 2016 ) and size of conidiomata or conidiogenous cells (Bezerra et al., 2019 ). Multigene-locus sequence phylogenies have also been used to distinguish different Pseudoplagiostoma species. The genetic markers employed in these studies were the internal transcribed spacer (ITS), large subunit of ribosomal DNA (LSU), RNA polymerase II ( RPB2 ), translational elongation factor EF-1α ( TEF ), and β-tubulin ( TUB ) (Bezerra et al., 2019 ; Cheewangkoon et al., 2010 ; Mu et al., 2022 ; Suwannarach et al., 2016 ; Tang et al., 2022 ; Zhang et al., 2023 ). Leaf spot disease has been found in many avocado orchards in Taiwan, and many growers and researchers think it might be an anthracnose disease and caused by Colletotrichum spp. However, in this study, a novel Pseudoplagiostoma species was isolated from diseased avocado leaves in Taiwan, and the pathogenicity of this new fungal species on avocado leaves was validated. Materials and methods Disease descriptions and fungal isolation An avocado leaf with leaf spots was photographed from October 16, 2019 to December 24, 2019 (Fig. 1 ). Initially, needle-like brown or black spots with a yellow halo appeared on the upper surface of the leaves (Fig. 1 A & 1 B). These needle-like spots gradually enlarged and became distinct, irregular, medium-brown lesions with dark-brown margins and black-brown centers on the upper surface (Fig. 1 C). Diseased samples were collected from April 2017 to November 2019 in five avocado orchards located in Chiayi [orchard 1: (23°29'08.1"N 120°28'11.4"E), orchard 2: (23°29'27.1"N 120°32'55.7"E), orchard 3: (23°24'58.4"N 120°30'44.3"E)], Chunghua (23°51'12.5"N 120°32'38.8"E), and Tainan Counties (23°05'45.4"N 120°21'42.1"E). Symptomatic leaves were kept in a sealed plastic box with moistened paper towels for 3 days to promote sporulation. Spores on the leaf lesions were collected and evenly spread on 2% water agar (WA) plates with a transferring loop. After spore germination, an agar disc with a single spore was cut from the WA and transferred to potato dextrose agar (PDA) (Merck, New Jersey, USA) to obtain axenic cultures. Morphology Colony morphology was investigated on PDA, malt extract agar (MEA) (Merck, New Jersey, USA), and oatmeal agar (OA) (Becton, Dickinson and Company, New Jersey, USA) after 7 days of incubation at 25 ℃ in darkness. Images of fungal structures (conidia, conidiogenous cells, asci, and ascospores) and measurements were made under a Nikon Eclipse Ni microscope (Nikon, Tokyo, Japan) with a ProgRess Gryphax NAOS camera and associated software (Jenoptik, Jena, Germany). The conidia and conidiogenous cells were observed on PDA after 21 days of incubation. To observe the sexual morph, the fungi were incubated on WA plates with sterilized Casuarina equisetifolia needles (Chuang et al., 2012 ) and observed after 21 days of incubation. In addition, to observe the acervuli and pycnidia on leaves, hand cross-sectioning of the fruiting bodies from leaf spots was performed. DNA template preparation, PCR amplification, and sequencing Ten isolates were used for molecular analysis, namely V14, V19, V22, V24, V25, V26, V27, V36, V58, and V64. For each isolate, mycelium was collected from the surface of PDA after 14 days of incubation at 25 ℃. The mycelium was ground in 0.5 N NaOH with an electronic grinder and centrifuged. The supernatant was mixed with 0.1 M Tris buffer (pH 8.0) in a ratio of 1:9 (v/v). The mixture was used as the DNA template for PCR. The primer pairs used to amplify the ITS, LSU, and TUB sequences were ITS1 and ITS4 (White et al., 1990 ), LR5 and LROR (Vilgalys & Hester 1990 ), and Bt1 (O’Donnell & Cigelnik 1997 ) and Bt2b (Glass & Donaldson 1995 ), respectively. Each 25 µL reaction mixture consisted of 0.2 µL (2 U) of Phusion High-fidelity DNA polymerase (ThermoFisher, USA), 1.5 µL of DNA template, 1 µL of 10 mM dNTPs, 5 µL of 5× Phusion HF Buffer, and 17.3 µL of sterilized double-distilled water. The PCR reactions were performed in a ThermoFisher SensoQuest Labcycler thermal cycler. The PCR program for all genes was 98 ℃ for 30 s, followed by 35 cycles of 98 ℃ for 10 s, the appropriate annealing temperature for 30 s, 72 ℃ for 1 to 1.5 min, and a final 10 min elongation at 72 ℃. The annealing temperature was 54 ℃ for the amplification of ITS, 52 ℃ for LSU, and 55 ℃ for TUB . The amplified products were sent to Tri-I Biotech Incorporation Company (Taipei, Taiwan) for sequencing, and the sequences for the isolates in this study were deposited in the GenBank database (Table 1 ). Table 1 Fungal isolates used in phylogenetic analysis and their GenBank accession numbers GenBank accession number b Species Isolate/strain a Host/substrate Location ITS LSU TUB Apoharknessia eucalypti CBS 142518 Eucalyptus pellita Malaysia MG934432 MN162172 MG934505 Apoharknessia eucalyptorum CBS 142519 Eucalyptus pellita Malaysia KY979752 KY979807 KY979919 Pseudoplagiostoma alsophilae SAUCC WZ0451* Alsophila spinulosa China OP810625 OP810631 OP828586 Pseudoplagiostoma alsophilae SAUCC WZ0452 Alsophila spinulosa China OP810626 OP810632 OP828587 Pseudoplagiostoma bambusae SAUCC 1206-4* Bambusoideae sp. China OP810629 OP810635 OP828590 Pseudoplagiostoma bambusae SAUCC 1206-6 Bambusoideae sp. China OP810630 OP810636 OP828591 Pseudoplagiostoma castaneae SAUCCmy0162* Castanea mollissima China MZ156982 MZ156985 MZ220325 Pseudoplagiostoma castaneae SAUCCmy0523 Castanea mollissima China MZ156983 MZ156986 MZ220326 Pseudoplagiostoma corymbiae CBS 132529* Corymbia sp. Australia JX069861 JX069845 - Pseudoplagiostoma corymbiicola CBS 145052* Corymbia citriodora Australia MK047425 MK047476 MK047577 Pseudoplagiostoma dipterocarpi TBRC 1895* Dipterocarpus tuberculatus Thailand KR994682 KR994683 - Pseudoplagiostoma dipterocarpicola MFLUCC 21–0142* Dipterocarpus sp. Thailand OM228844 OM228842 OM219638 Pseudoplagiostoma dipterocarpicola MFLUCC 21–0114 Dipterocarpus sp. Thailand OM228843 OM228841 OM219637 Pseudoplagiostoma eucalypti CBS 124807* Eucalyptus urophylla Venezuela GU973512 GU973606 GU973575 Pseudoplagiostoma eucalypti CPC 14161 Eucalyptus camaldulensis Vietnam GU973510 GU973604 GU973573 Pseudoplagiostoma eucalypti CPC 12280 Eucalyptus sp. USA GU973507 GU973601 GU973570 Pseudoplagiostoma eucalypti CPC 115743 Eucalyptus globulus Uruguay GU973509 GU973603 GU973572 Pseudoplagiostoma inthanonense MFLUCC 23–0262 Plant litter Thailand OR606510 OR633320 OR611920 Pseudoplagiostoma mangiferae KUMCC 18–0179* Mangifera sp. China MK084824 MK084825 - Pseudoplagiostoma myracrodruonis URM 7799* Astronium urundeuva Brazil MG870421 MK982151 MN019566 Pseudoplagiostoma myracrodruonis URM 8123 Astronium urundeuva Brazil MK982150 MK982152 MN019567 Pseudoplagiostoma machili SAUCC BW0233* Machilus nanmu China OP810627 OP810633 OP828588 Pseudoplagiostoma machili SAUCC BW0221 Machilus nanmu China OP810628 OP810634 OP828589 Pseudoplagiostoma jasmini MFLUCC 23–0044 Jasminum grandiflorum Thailand OQ786078 OQ786079 OQ850148 Pseudoplagiostoma oldii CBS 115722 Eucalyptus camaldulensis Australia GU973535 GU973610 GU993864 Pseudoplagiostoma oldii CBS 124808* Eucalyptus camaldulensis Australia GU973534 GU973609 GU993862 Pseudoplagiostoma perseae V14; BCRC FU31382 Persea americana Taiwan MT233353 MT233367 MT251137 Pseudoplagiostoma perseae V19; BCRC FU31383 Persea americana Taiwan MT233354 MT233368 MT251138 Pseudoplagiostoma perseae V22; BCRC FU31384 Persea americana Taiwan MT233355 MT233369 MT251139 Pseudoplagiostoma perseae V24; BCRC FU31385 Persea americana Taiwan MT233356 MT233370 MT251140 Pseudoplagiostoma perseae V25; BCRC FU31386 Persea americana Taiwan MT233357 MT233371 MT251141 Pseudoplagiostoma perseae V26; BCRC FU31387 Persea americana Taiwan MT233358 MT233372 MT251142 Pseudoplagiostoma perseae V27; BCRC FU31388* Persea americana Taiwan MT233359 MT233373 MT251143 Pseudoplagiostoma perseae V36; BCRC FU31389 Persea americana Taiwan MT233360 MT233374 MT251144 Pseudoplagiostoma perseae V58; BCRC FU31390 Persea americana Taiwan MT233361 MT233375 MT251145 Pseudoplagiostoma perseae V64; BCRC FU31391 Persea americana Taiwan MT233362 MT233376 MT251146 Pseudoplagiostoma variabile CBS 113067* Eucalyptus globulus Uruguay GU973536 GU973611 GU993863 a Ex-type strains are marked with “*”. b ITS = internal transcribed spacer; LSU = large subunit of ribosomal DNA; and TUB = β-tubulin-2. Phylogenetic analysis The three nuclear gene regions used in phylogenetic analysis were ITS, LSU, and TUB . Sequence data of 27 Pseudoplagiostoma isolates were retrieved from the GenBank database (Table 1 ). Multiple sequence alignment of each gene was conducted using ClustalX v. 2.1 (Larkin et al., 2007 ) and these alignments were concatenated using Sequence Matrix v. 1.7.8 (Vaidya et al., 2011 ). The best DNA substitution model for each gene region was predicted by jModeltest 2.1.7 (Posada, 2008 ). The DNA substitution models used for ITS, LSU, and TUB were SYM + G, TrNef + I, and HKY + I, respectively. A Bayesian phylogenetic tree was constructed with the software Mr. Bayes v. 3.2.6 (Ronquist et al., 2012 ). The analysis was run twice with 1×10 7 generations, and samples were taken from the posterior probability trees every 1,000 generations. The first 25% of generations were discarded as burn-in. Apoharknessia eucalypti CBS 142518 and Apoharknessia eucalyptorum CBS 142519 were used as the outgroups. A maximum Likelihood (ML) tree was constructed using IQ tree 2.1.3 (Nguyen et al., 2015 ) with 1,000 bootstrap replicate. The matrices and phylogenetic trees were deposited in the TreeBASE database ( http://purl.org/phylo/treebase/phylows/study/TB2:S31183 ). Pathogenicity tests One-year-old avocado seedlings of the ‘Choquette,’ ‘Hall,’ and ‘Hung Shin Yuan’ cultivars were used in the pathogenicity tests using the Pseudoplagiostoma isolate V27. For each cultivar, three plants were used in one pathogenicity test, and for each plant, two leaves were inoculated. Conidia were harvested from PDA culture plates after 21 days of incubation at 25 ℃ in the dark. Conidia were diluted with the appropriate amount of sterilized distilled water to obtain a concentration of 2×10 4 spores/ml. The spore suspension was sprayed on both the upper and lower sides of young avocado leaves until runoff. The inoculated leaves were sealed in zipper bags with moistened tissue paper to maintain high humidity. Leaves sprayed with sterile distilled water were used as the negative control. The inoculated leaves were removed from the zipper bags 2 days after inoculation and kept in a greenhouse with ambient light and temperature. The pathogenicity test was conducted twice. The first trial was conducted from October 18 to November 1, 2023. The mean temperature during this period was 24.91 ℃, with 8.81 h of sunshine per day on average. The second trial was conducted from November 1 to November 15, 2023. The mean temperature during this period was 23.99 ℃, with 8.83 h of sunshine per day. For the inoculation tests on the three cultivars, leaves of inoculated seedlings were observed 2 weeks after inoculation. To fulfill Koch’s postulates, the fungi were recovered from the spots on the leaves and identified based on morphological traits and partial TUB sequences. For the fruit inoculation assay, Pseudoplagiostoma isolate V27 was incubated on PDA agar at 25 ℃ for 7 days. An agar disc 5 mm of diameter was cut from the edge of the fungal colony and placed on a wound on avocado fruit made using a needle (23G × 1.25″). After inoculation, the avocado fruits were placed in a basin, and the basin wrapped in a plastic bag for 2 days. The inoculated fruits were stored at 25 ℃ in darkness. The inoculated fruits were investigated after 7 days of inoculation. Results Field symptoms and fungal isolation Sixty-three isolates were obtained from avocado leaf spots in different avocado orchards. All 63 isolates exhibited similar colony morphologies viewed on the surface and reverse sides of OA (Fig. 2 A, D), MEA (Fig. 2 B, E), and PDA (Fig. 2 C, F) plates. The surface of the colony appeared dark gray, gray to grayish, and olive green (Fig. 2 A, B, C), and the colony appeared dark gray to olive green when viewed from the reverse side of the plate (Fig. 2 D, E, F). Ten isolates were chosen arbitrarily to conduct the subsequent phylogenetic analysis. Phylogenetic analysis A Bayesian inference phylogenetic tree based on ITS, LSU, and TUB sequences revealed that the ten P. perseae isolates (V14, V19, V22, V24, V25, V26, V27, V36, V58, V64) formed an independent clade, which was sister to that of Pseudoplagiostoma alsophilae and Pseudoplagiostoma machili (Fig. 3 ). The clade was supported by a Bayesian posterior probability of 100% and ML bootstrap value of 100%. Taxonomy Pseudoplagiostoma perseae C. J. Wu, J. L. Chen, S. S. Tzean, H. F. Ni, sp. nov. [Mycobank No:] MB839192 (Figs. 2 and 4 ). Etymology In reference to the host genus name, Persea . Typification Beidou, Changhua, Taiwan (23.853461, 120.544106), isolated from a diseased avocado leaf. Ex-type living culture is BCRC FU31388 (V27) (Holotype TNM F0037701). Several other paratype cultures isolated from different locations and diseased leaf samples were maintained via cryopreservation in the Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan (BCRC FU31382–BCRC FU31387 and BCRC FU31389–BCRC FU31391). The GenBank accession numbers of the ten P. perseae isolates are shown in Table 1 . Description On leaves, Conidiomata acervular, subepidermal, light brown, 107.39 to 205.67 µm in width, 55.51 to 92.62 µm high (145.21 ± 31.41 × 75.67 ± 11.88 µm, n = 15), often with pale yellow droplet exudates containing masses of conidia on the leaf surface. Thin conidiomata wall, consisting of one to two layers of textura angularis ; dehiscence irregular, by rupture of the host epidermal tissue. Conidiophores absent. Conidiogenous cells enteroblastic, annellidic, discrete, indeterminate, ovoid, obclavate or obpyriform, smooth, subhyaline to pale brown, with several percurrent proliferations in the apical part, 8.61–21.68 × 5.86–14.93 µm (14.24 ± 3.52 × 9.02 ± 2.57 µm, n = 25). Conidia 17.66–23.68 × 10.62–16.48 µm (20.15 ± 1.35 × 13.20 ± 1.31 µm, n = 50), acrogenous, ellipsoidal, mostly aseptate, but rarely with one to three septa, smooth, hyaline to pale yellowish grey or pale brown, with a protruding scar at the base. On leaves, ascomata globose, subglobose or irregular, intra- or subepidermal; wall consisting of two to three layers of dark brown to black textura angularis . Ostioles beaked lateral, neck internally lateral lined with periphysis. Paraphyses absent. Asci unitunicate, elongated clavate or fusiform with blunt or rounded apex, sessile, 8-spored, hyaline. Ascospores fusiform, one-septate, two-celled unequally, upper cell longer and wider, hyaline, smooth, constricted at the septum, with appendage at both ends, 26.15–30.22 × 12.39–14.94 µm (28.07 ± 0.15 × 13.60 ± 0.071 µm, n = 50, on C. equisetifolia needles). Culture characteristics On PDA, colonies reached 4.4 cm in diameter after a week of incubation at 25 ℃ in darkness. Surface floccose to felted, gray to grayish green. Reverse dark gray to olive green. The colony features on OA, MEA, and PDA are shown in Fig. 2 . Notes A phylogenetic tree based on ITS, LSU, and TUB gene markers showed that P. perseae formed a well-supported lineage with a high posterior probability (100% from Bayesian inference) and bootstrap value (100% from ML analysis) (Fig. 3 ). P. perseae differed from all other Pseudoplagiostoma spp. in terms of its conidia size (18–24 × 12–14 µm) and shape except for P. alsophilae (17–21 × 13–15 µm), P . machili (17.5–23 × 10.5–13.5 µm), and P. mangiferae (18–24 × 11–14 µm). However, the conidiomata of P. alsophilae , P. machili , and P. mangiferae were pycnidial, whereas P. perseae had acervuli. The colony surfaces of both P. alsophilae and P. machili on PDA were gray white to creamy white with irregular margins, and the reverse sides were similar (Zhang et al., 2023 ). The surface of the P. mangiferae colony on PDA was pink-white to cream and pale yellowish when viewed from the reverse side (Phookamsak et al., 2019 ). By contrast, most P. perseae colonies were gray to grayish olive green on the surface and dark gray to olive green when viewed from the reverse side. The differences in morphology and phylogenetic relatedness indicated that P. perseae is a new species. The hosts and characteristics of the conidiomata, conidiogenous cells, and conidia of the 16 currently known Pseudoplasgiostoma species are shown in Table 2 . Table 2 Asexual morphological features of Pseudoplagiostoma species Species Host a Conidiomata type and size (in µm) Conidiogenous cell size (in µm) Conidia size (in µm) References P. alsophilae Alsophila spinulosa Pycnidial, 150–250 × 200–300 8–13 × 1.5–3 17–21 × 13–15 Zhang et al., 2023 P. bambusae Bambusoideae Pycnidial, 200–250 × 150–250 5–13 × 1.5–2.5 13–20 × 5.7–7.6 Zhang et al., 2023 P. castaneae Castanea mollissima Pycnidial, 270–450 × 220 − 400 8–35 × 1–2 9–13.5 × 2–4.5 Mu et al., 2022 P. corymbiae Corymbia sp. (‎Myrtaceae) Acervular, up to 300 diameter 10–20 × 4–7 (14–)16–18(–19) × (7–)8–9(–10) Crous et al., 2012 P. corymbiicola Corymbia citriodora , Corymbia variegata (‎Myrtaceae) Acervular, up to 300 diameter 15–30 × 3–5 (15–)16–17(–20) × (6–)7(–8) Crous et al., 2018 ; Crous et al., 2019 P. dipterocarpi Dipterocarpus tuberculatus (Dipterocarpaceae) – 18–25 × 2.5–4.5 14–36 × 7–11 Suwannarach et al., 2016 P. dipterocarpicola Dipterocarpus sp. Pycnidial, 63–153 × 113–288 5.0–11 × 1.0–2.5 9.0–22 × 4.0–7.5 Tang et al., 2022 P. eucalypti Eucalyptus camaldulensis , Eucalyptus grandis , Eucalyptus robusta , Eucalyptus urophylla ( Myrtaceae) Acervular to pycnidial, (170–)180–200(–230) wide and (150–)170–190(–220) high (6–)8–12(–15) × 2–4(–6) (15–)17–19(–23) × (6.5–)7–8(–8.5); on MEA, (14–)16–19(–22) × (6–)7–9(–11) Cheewangkoon et al., 2010 ; Fan et al., 2018; Wang et al., 2016 P. inthanonense Plant litter Pycnidial, 80–140 height, 70–120 diameter (103 × 109) 3.3–12.3 × 1–2.7 ( 7 × 1.7) 14–22.6 × 6.4–10.2 (17.5 − 8.5 µm) Hittanadurage et al., 2023 P. machili Machilus nanmu Pycnidial, 150–200 × 100–250 7–16 × 2–3.5 17.5–23 × 10.5–13.5 Zhang et a. 2023 P. mangiferae Mangifera sp. Pycnidial, 70–140 high and 90–150 diameter 5–11 × 3.2–12.6 18–24 × 11–14 Phookamsak et al., 2019 P. myracrodruonis Myracrodruon urundeuva Pycnidial, 250–290 × 236–245 7–7.5 × 2–3(–3.5) (10–)12–15(–19) × (4–)5–6(–7.5) Bezerra et al., 2019 P. jasmini Jasminum grandiflorum Pycnidial, (135–)140–200 high, (145–)150–230(–240) diameter (mean = 184 × 171) (6.8–)7.7–13.7(–15.6) × 1.6–2.4(–3.0) (mean = 10.7 × 2.1) (11.8–)14–22 × (5.2–)6.5–11 (mean = 18.5 × 9.5) Gomdola et al., 2023 P. oldii Eucalyptus camaldulensis ( Myrtaceae) Acervular to pycnidial, (265–)285–300(–330) wide and (200–)220–250(–270) high (8.5–)15–20(–26) × 2–3(–4.5) (15–)17–20(–23) × (6–)7–8(–9); on MEA, (11–)14–17(–20) × (6–)7–9(–11) Cheewangkoon et al., 2010 P. perseae Persea americana Acervular, 107.39–205.67 wide and 55.51–92.62 high 8.61–21.68 × 5.86–14.93 17.82–24.15 × 11.73–15.57; on MEA 16.61–21.73 × 12.11–14.92 This study P. variabile Eucalyptus globulus ( Myrtaceae) Pycnidial, (145–)170–190(–245) wide and (130–)160–180(–230) high (12–)15–20(–23) × 2–3(–4.5) (12.5–)15.5–17.5(–23.5) × (5.5–)6.5–8(–9); on MEA, (6.5–)15.5–17(–19) × (6.5–)7.5–9(–10.5) Cheewangkoon et al., 2010 a Host: host Sensu lato, including endophyte hosts. Pathogenicity tests In pathogenicity tests, P. perseae caused leaf spots on the ‘Choquette,’ ‘Hall,’ and ‘Hung Shin Yuan’ cultivars 2 weeks post-inoculation. Needle-like black spots appeared on leaves (Fig. 5 A-D). Some lesions developed blurry halos (Fig. 5 A, C). The spots also appeared on veins on the reverse sides of leaves (Fig. 5 B). The leaves of the negative control (Fig. 5 E), which was inoculated with sterilized water, had no symptoms. To fulfill Koch’s postulates, spots on inoculated leaves were cut, sterilized, and incubated on PDA plates, and P. perseae were identified according to colony morphology and partial TUB sequence. P. perseae were re-isolated from all inoculated leaves, thus fulfilled Koch’s postulates. Fruit pathogenicity tests were conducted by placing agar discs bearing P. perseae mycelium on wounded fruits. No symptoms were visible after one week of observation of avocado fruits (data not shown). Discussion This is the first report characterizing a leaf spot disease on avocado leaves, and the causal agent of this disease, P. perseae is a new species according to analysis of colony morphology, spore size, sporulation structure, host preference, and phylogenetic analysis with ITS, LSU, and TUB gene markers. In pathogenicity tests, P. perseae was pathogenic on the leaves of the three avocado cultivars tested. The ITS, LSU, and TUB loci are commonly used to distinguish species under the genus Pseudoplagiostoma . For example, Cheewangkoon et al., ( 2010 ) concatenated ITS and TUB to distinguish P. eucalypti , P. oldii , and P. variable . Many studies of Pseudoplagiostoma species also included ITS, LSU, and TUB in their phylogenetic analyses (Bezerra et al., 2019 ; Crous et al., 2018 ; Gomdola et al., 2023 ; Hittanadurage et al., 2023; Mu et al., 2022 ; Phookamsak et al., 2019 ; Suwannarach et al., 2016 ; Zhang et al., 2023 ). Other gene markers, such as TEF and RPB2 , are also used to resolve Pseudoplagiostoma spp. (Bezerra et al., 2019 ; Hittanadurage et al., 2023; Mu et al., 2022 ; Tang et al., 2022 ; Zhang et al., 2023 ). However, for P. corymbiae , P. corymbiicola , P. dipterocarpi , and P. diptercarpicola , no RPB2 sequences have been deposited in GenBank or other DNA sequence databases; thus, using RBP2 to distinguish Pseudoplagiostoma spp. might be problematic. Colletotrichum spp. might have been mistaken as the causal agent of avocado leaf spot disease in field diagnosis in the past. This is mainly because some Colletotrichum spp. are known pathogens and endophytes in avocados (Giblin et al., 2010 ; Shetty et al., 2016 ). Use of the appropriate methods and timing are crucial to isolate P. perseae from symptomatic leaves. In this study, leaves with lesions were kept in a moistened box to induce sporulation of P. perseae . Using the general fungal isolation method, which involves disinfecting the leaf surface and incubating the leaf discs on a PDA plate, the fungal species regenerated from the disease tissue might be Colletotrichum spp. most of the time, since Colletotrichum spp. grow fast and are common endophytes, pathogens, and saprophytes on avocados. However, the chance of P. perseae being isolated from leaf tissues increases greatly if isolation is performed at an incipient infection stage, i.e., when needle-like black spots have just appeared on the avocado leaves. At least eight species in the Pseudoplagiostoma genus are associated with foliar disease. The pathogenicities of P. eucalypti and P. mangiferae have been validated on eucalyptus and mango, respectively (Cheewangkoon et al., 2010 ; Sankaran et al., 1995 ; Zhou et al., 2022 ), whereas P. corymbiae and P. corymbiicola were isolated from leaf spots of the lemon-scented gum tree ( Corymbia spp.) (Crous et al., 2018 , 2019 ). P. alsophilae , P. bambusae , P. castaneae , P. jasmini , and P. machili were also isolated from leaf spots on their preferred host plants, e.g., bamboo or chestnut (Mu et al., 2022 ; Zhang et al., 2023 ). In addition, P. dipterocarpi and P. myracroduonis are endophytic, living in the leaves of tree species such as the Indian English gurjun tree ( Dipterocarpus tuberculatus ) and Urunday tree ( Myracrodruon urundeuva ) (Bezerra et al., 2019 ; Suwannarach et al., 2016 ). Pseudoplagiostoma dipterocarpicola was isolated from dead twigs and fruits of Dipterocarpus sp. (Keruing). P. inthanonense was isolated from plant litter and might be a saprophyte (Hittanadurage et al., 2023). In Taiwan, three Pseudoplagiostoma spp. have already been documented and validated to be pathogenic on their respective hosts: P. eucalypti , which causes leaf spots on swamp mahogany ( Eucalyptus robusta ) (Wang et al., 2016 ), P. mangiferae , the causal agent of mango leaf blotch (Zhou et al., 2022 )d perseae , which causes avocado leaf spot, as demonstrated in this study. In the future, the host range, pathogenicity on different avocado cultivars, etiology, and fungicide sensitivity of P. perseae will be further studied. In planta field assays should also be conducted to confirm the effectiveness of fungicides, which have been tested against P. perseae in vitro . The information gained from these studies will help avocado growers to control P. perseae , which causes leaf spot disease of avocado. This will in turn benefit the avocado industry. Declarations Statements and Declarations Compliance with Ethical Standards There are no potential conflicts of interest. This research is not involving Human Participants and/or Animals. Therefore, there is no informed consent needed. Competing Interests The authors have no relevant financial or non-financial interests to disclose. References Bezerra, J. D. P., Pádua, A. P. S. L., Oliveira, T. G. L., Paiva, L. M., Guarnaccia, V., Fan, X., & Souza-Motta, C. M. (2019). Pseudoplagiostoma myracrodruonis ( Pseudoplagiostomataceae , Diaporthales ): a new endophytic species from Brazil. Mycological Progress , 18, 1329–1339. Cheewangkoon, R., Groenewald, J. Z., Verkley, G. J. M., Hyde, K. D., Winfield, M. J., Gryzenhout, M., Summerell, B. A., Denman, S., Toanun, C., & Crous, P. W. (2010). Re-evaluation of Cryptosporiopsis eucalypti and Cryptosporiopsis -like species occurring on Eucalyptus leaves. Fungal Diversity , 44 , 89–105. Crous, P. W., Luangsa-Ard, J. J., Wingfield, M. J., Carnegie, A. J., Hernández-Restrepo, M., Lombard, L., Roux, J., Barreto, R. (2018). Fungal Planet description sheets: 785– 867. 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E. (1992). Avocado diseases. Florida Agricultural Experiment Station Bulletin 161. Florida . Agricultural Experiment Station, University of Florida. Suwannarach, N., Kumla, J., & Lumyong, S. (2016). Pseudoplagiostoma dipterocarpi sp. nov., a new endophytic fungus from Thailand. Mycoscience , 57 , 118–122. Tang, X., Jayawardena, R., Stephenson, S., & Kang, J. C. (2022). A new species Pseudoplagiostoma dipterocarpicola ( Pseudoplagiostomataceae , Diaporthales ) found in northern Thailand on members of the Dipterocarpaceae . Phytotaxa , 543 , 233–243. Vaidya, G., Lohman, D. J., & Meier, R. (2011). SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics , 27 , 171–180. Vilgalys, R., & Hester, M. (1990). Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology , 172 , 4238–4246. Wang, C. L., Yang, S. W., & Chiang, C. Y. (2016). The first report of leaf spot of Eucalyptus robusta caused by Pseudoplagiostoma eucalypti in Taiwan. Plant Disease , 100 , 1504. White, T. J., Bruns, T., Lee, S., & Taylor, J. W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In M. A. Innis, D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR Protocols: A Guide to Methods and Applications (pp. 315–322). Academic.). Zentmyer, G. A. (1953). Disease of the avocado (pp. 875–881). United States Department of Agriculture Yearbook. Zhang, Z., Liu, X., Tao, M., Liu, X., Xia, J., Zhang, X., & Meng, Z. (2023). Taxonomy, phylogeny, divergence time estimation, and biogeography of the family Pseudoplagiostomataceae ( Ascomycota , Diaporthales ). Journal of Fungi , 9, 82. Zhou, Z. Y., Tsao, W. C., Chung, W. H., & Wang, C. L. (2022). First report of mango leaf blotch caused by Pseudoplagiostoma mangiferae in Taiwan. Plant Disease , 106 , 2749. Cite Share Download PDF Status: Published Journal Publication published 01 Aug, 2024 Read the published version in European Journal of Plant Pathology → Version 1 posted Editorial decision: Revision 07 May, 2024 Reviewers agreed at journal 10 Mar, 2024 Reviewers invited by journal 07 Mar, 2024 Editor invited by journal 07 Mar, 2024 Editor assigned by journal 06 Mar, 2024 First submitted to journal 05 Mar, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4019244","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":276922186,"identity":"e92a8815-15a0-4efe-a7d3-04b78e220c1a","order_by":0,"name":"Chao-jung Wu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3klEQVRIiWNgGAWjYJAC5j88Ejz8IFYCMcp5wISMhZxkG1wLMzFabCqMDY4hbMWvxZ797MEbEjkSiZvvNz/78IChVs7gAP8xCby28OQlWxickUjcdozNeEYCw3FjgwPMbPi1SPCYSST2gLQwGAP9cixxZgMz2w2CWg7+Azqsjf0z8VokG3gkjA3YeEC21CT2MxDSciYv2RqoUU7iWE4xQ4LBAWN+ZmbzH/i0sLefPXibgaeOh7/5+GbGHxV1cmzsjY8N8GkBxQxS+BgcJhgt6FoY6giqHwWjYBSMgpEHAPRJPQBbnRfPAAAAAElFTkSuQmCC","orcid":"","institution":"Taiwan Agricultural Research Institute Chiayi Agricultural Experiment Branch","correspondingAuthor":true,"prefix":"","firstName":"Chao-jung","middleName":"","lastName":"Wu","suffix":""},{"id":276922187,"identity":"5e8f9172-8eaf-4b32-9949-80e9c13be9cf","order_by":1,"name":"Jin Liang Chen","email":"","orcid":"","institution":"Chia Nan University of Pharmacy and Science","correspondingAuthor":false,"prefix":"","firstName":"Jin","middleName":"Liang","lastName":"Chen","suffix":""},{"id":276922188,"identity":"149cdd97-a4f7-482c-97d3-dbd460c5ce5a","order_by":2,"name":"Shean Shong Tzean","email":"","orcid":"","institution":"National Taiwan University","correspondingAuthor":false,"prefix":"","firstName":"Shean","middleName":"Shong","lastName":"Tzean","suffix":""},{"id":276922189,"identity":"390d91ac-64c1-4eff-9db7-7a1ae63ffe1d","order_by":3,"name":"Hui Fang Ni","email":"","orcid":"","institution":"Taiwan Agricultural Research Institute Chiayi Agricultural Experiment Branch","correspondingAuthor":false,"prefix":"","firstName":"Hui","middleName":"Fang","lastName":"Ni","suffix":""}],"badges":[],"createdAt":"2024-03-06 04:12:46","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4019244/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4019244/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10658-024-02921-1","type":"published","date":"2024-08-01T15:57:09+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":52453381,"identity":"9a0643ad-3699-44a4-b66b-4cf8a8a4e355","added_by":"auto","created_at":"2024-03-11 19:24:13","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3571816,"visible":true,"origin":"","legend":"\u003cp\u003eLeaf spot symptoms observed in the field. (A) A leaf with needle-like spots with a yellow halo appeared on adaxial surface during the incipient stage on October 16, 2019. (B) Symptoms on the same leaf on November 20, 2019. (C) Brown lesions observed on the leaf on December 24, 2019\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/b1d38fe887b8aa9320d1ca19.png"},{"id":52453380,"identity":"d34d04bf-5827-4bf6-bede-6fa9b8be1e5a","added_by":"auto","created_at":"2024-03-11 19:24:13","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":5406031,"visible":true,"origin":"","legend":"\u003cp\u003eColony, anamorph, and teleomorph of \u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e isolate V27. The surface and reverse sides of the culture on oatmeal agar (A, D), malt extract agar (B, E), and potato dextrose agar (PDA) (C, F). Anamorph: (G) incipient acervulus, (H) mature acervulus, (I) conidia mass, (J)-(K) conidiogenous cells with conidia at different developmental stages. (L) - (P) Teleomorph images: (L) ascospores, (M) - (N) an ascus, (O) perithecium, with the whole global structure shown, and (P) longitudinal section of a perithecium neck with periphysis. (G, H, O, P) images of \u003cem\u003eP. perseae \u003c/em\u003eon an avocado leaf; (I)-(K), images on PDA; (L)-(N), images on \u003cem\u003eCasuarina equisetifolia\u003c/em\u003e needles. (H, O, P) share the same scale bar as (G), which is 50 μm. (J) - (N) share the same bar as (I), which is 20 μm\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/019e46145f1b0aad42d683a1.png"},{"id":52453378,"identity":"18f7c08a-d4e9-4026-8fb9-4295f9286fd2","added_by":"auto","created_at":"2024-03-11 19:24:13","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1630875,"visible":true,"origin":"","legend":"\u003cp\u003eBayesian inference phylogenetic tree of \u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e and other \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species based on ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e sequences. \u003cem\u003eApoharknessia eucalyptorum\u003c/em\u003e CBS 142519 and \u003cem\u003eApoharknessia eucalypti\u003c/em\u003e CBS 142518 served as the outgroups. The Bayesian posterior probability (PP) values above 0.95 and maximum likelihood bootstrap support (BS) values above 75% are shown at the nodes (PP/BS). Ex-type strains are emphasized in bold. The scale bar indicates the number of expected changes per site\u003c/p\u003e","description":"","filename":"Fig3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/ec42eb0f2cb75e20df0ee813.jpg"},{"id":52453379,"identity":"4a22499d-8392-4800-8a78-a08ab662cdf9","added_by":"auto","created_at":"2024-03-11 19:24:13","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2228115,"visible":true,"origin":"","legend":"\u003cp\u003eIllustration of the acervulusof \u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e. Conidiogenous cells and conidia are shown. Scale bar = 10 μm\u003c/p\u003e","description":"","filename":"Fig4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/2a3993d5c01a2871f1150e23.jpg"},{"id":52453383,"identity":"f01eba5d-0f18-4063-8616-55646c4d7471","added_by":"auto","created_at":"2024-03-11 19:24:13","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":4862738,"visible":true,"origin":"","legend":"\u003cp\u003eSymptoms on the leaves of different avocado cultivars 2 weeks post-inoculation. (A) An inoculated leaf of ‘Choquette.’ (B) The reverse side of an inoculated ‘Choquette’ leaf. (C) An inoculated leaf of ‘Hall.’ (D) An inoculated leaf of ‘Hung Shin Yuan.’ (E) A ‘Choquette’ leaf inoculated with sterilized water as a negative control\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/450c2161a16b1b726bf83be5.png"},{"id":61793909,"identity":"185828bf-f52e-418e-b146-89298fc8129f","added_by":"auto","created_at":"2024-08-05 16:16:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":26924266,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4019244/v1/c56304fa-269a-469f-a7f4-ed901f0ad57d.pdf"}],"financialInterests":"","formattedTitle":"Pseudoplagiostoma perseae sp. nov. causes leaf spot disease on avocado leaves in Taiwan","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAvocado is one of the most popular tropical fruit crops in the world. According to the Food and Agriculture Organization (FAO) of the United Nations, 2.4\u0026nbsp;million tons of avocado were exported globally in 2022 (FAO \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In Taiwan, the avocado planting area was about 1,811 hectares in 2022, which represents a 147% increase from the planting area in 2016 (733 hectares). Some cultivars, such as \u0026lsquo;Changan,\u0026rsquo; Choquette,\u0026rsquo; \u0026lsquo;Hall,\u0026rsquo; and \u0026lsquo;Hung Shin Yuan\u0026rsquo; are commonly planted in orchards.\u003c/p\u003e \u003cp\u003eThere are three main pathogens known to cause lesions on avocado leaves. \u003cem\u003eCercospora purpurea\u003c/em\u003e (Pohronezny et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1994\u003c/span\u003e) causes \u003cem\u003eCercospora\u003c/em\u003e spot or blotch disease, which is initially characterized by small light-brown spots appearing on leaves, which later turn into angular brown lesions with a small yellow halo. Sometimes greyish fruiting bodies are visible in lesion centers under high humidity (Ruehle \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1943\u003c/span\u003e; Stevens \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1992\u003c/span\u003e; Zentmyer \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1953\u003c/span\u003e). \u003cem\u003eColletotrichum\u003c/em\u003e species are also pathogenic on avocado leaves and cause symptoms including brown spots randomly spread on the tips of senescing leaves (Giblin et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Pohronezny et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1994\u003c/span\u003e). \u003cem\u003eSphaceloma perseae\u003c/em\u003e causes avocado scab on fruits as well as leaves (Jenkins \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1925\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1934\u003c/span\u003e). The lesions on avocado leaves are grayish, oval to elongated in shape, and slightly elevated; they are mainly found on the reverse side of the leaf and mostly confined to the midrib and veins (Pegg et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). So far, no avocado leaf disease caused by \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species has been reported.\u003c/p\u003e \u003cp\u003eCurrently, the genus \u003cem\u003ePseudoplagiostoma\u003c/em\u003e contains 15 species (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Cheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Crous et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2012\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Gomdola et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Hittanadurage et al., 2023; Mu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Phookamsak et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Suwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Tang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The asexual structures of the genus \u003cem\u003ePseudoplagiostoma\u003c/em\u003e include acervular to pycnidial conidiomata with cylindrical to ampulliform conidiogenous cells and aseptate ellipsoidal conidia; the sexual structures are depressed globose or elliptical ostiolate perithecia with subcylindrical to long obovoid asci, which bear eight, two seriate, hyaline, medianly one-septate ellipsoidal ascospores with tapering ends and terminal elongate appendages (Cheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species are distinguished using morphological characteristics of the conidia (size, color, and shape) (Suwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and size of conidiomata or conidiogenous cells (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Multigene-locus sequence phylogenies have also been used to distinguish different \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species. The genetic markers employed in these studies were the internal transcribed spacer (ITS), large subunit of ribosomal DNA (LSU), RNA polymerase II (\u003cem\u003eRPB2\u003c/em\u003e), translational elongation factor EF-1α (\u003cem\u003eTEF\u003c/em\u003e), and β-tubulin (\u003cem\u003eTUB\u003c/em\u003e) (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Cheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Mu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Suwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Tang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLeaf spot disease has been found in many avocado orchards in Taiwan, and many growers and researchers think it might be an anthracnose disease and caused by \u003cem\u003eColletotrichum\u003c/em\u003e spp. However, in this study, a novel \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species was isolated from diseased avocado leaves in Taiwan, and the pathogenicity of this new fungal species on avocado leaves was validated.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eDisease descriptions and fungal isolation\u003c/h2\u003e \u003cp\u003eAn avocado leaf with leaf spots was photographed from October 16, 2019 to December 24, 2019 (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Initially, needle-like brown or black spots with a yellow halo appeared on the upper surface of the leaves (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA \u0026amp; \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). These needle-like spots gradually enlarged and became distinct, irregular, medium-brown lesions with dark-brown margins and black-brown centers on the upper surface (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eDiseased samples were collected from April 2017 to November 2019 in five avocado orchards located in Chiayi [orchard 1: (23\u0026deg;29'08.1\"N 120\u0026deg;28'11.4\"E), orchard 2: (23\u0026deg;29'27.1\"N 120\u0026deg;32'55.7\"E), orchard 3: (23\u0026deg;24'58.4\"N 120\u0026deg;30'44.3\"E)], Chunghua (23\u0026deg;51'12.5\"N 120\u0026deg;32'38.8\"E), and Tainan Counties (23\u0026deg;05'45.4\"N 120\u0026deg;21'42.1\"E). Symptomatic leaves were kept in a sealed plastic box with moistened paper towels for 3 days to promote sporulation. Spores on the leaf lesions were collected and evenly spread on 2% water agar (WA) plates with a transferring loop. After spore germination, an agar disc with a single spore was cut from the WA and transferred to potato dextrose agar (PDA) (Merck, New Jersey, USA) to obtain axenic cultures.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eMorphology\u003c/h2\u003e \u003cp\u003eColony morphology was investigated on PDA, malt extract agar (MEA) (Merck, New Jersey, USA), and oatmeal agar (OA) (Becton, Dickinson and Company, New Jersey, USA) after 7 days of incubation at 25 ℃ in darkness. Images of fungal structures (conidia, conidiogenous cells, asci, and ascospores) and measurements were made under a Nikon Eclipse Ni microscope (Nikon, Tokyo, Japan) with a ProgRess Gryphax NAOS camera and associated software (Jenoptik, Jena, Germany). The conidia and conidiogenous cells were observed on PDA after 21 days of incubation. To observe the sexual morph, the fungi were incubated on WA plates with sterilized \u003cem\u003eCasuarina equisetifolia\u003c/em\u003e needles (Chuang et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and observed after 21 days of incubation. In addition, to observe the acervuli and pycnidia on leaves, hand cross-sectioning of the fruiting bodies from leaf spots was performed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eDNA template preparation, PCR amplification, and sequencing\u003c/h2\u003e \u003cp\u003eTen isolates were used for molecular analysis, namely V14, V19, V22, V24, V25, V26, V27, V36, V58, and V64. For each isolate, mycelium was collected from the surface of PDA after 14 days of incubation at 25 ℃. The mycelium was ground in 0.5 N NaOH with an electronic grinder and centrifuged. The supernatant was mixed with 0.1 M Tris buffer (pH 8.0) in a ratio of 1:9 (v/v). The mixture was used as the DNA template for PCR. The primer pairs used to amplify the ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e sequences were ITS1 and ITS4 (White et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1990\u003c/span\u003e), LR5 and LROR (Vilgalys \u0026amp; Hester \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1990\u003c/span\u003e), and Bt1 (O\u0026rsquo;Donnell \u0026amp; Cigelnik \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1997\u003c/span\u003e) and Bt2b (Glass \u0026amp; Donaldson \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1995\u003c/span\u003e), respectively. Each 25 \u0026micro;L reaction mixture consisted of 0.2 \u0026micro;L (2 U) of Phusion High-fidelity DNA polymerase (ThermoFisher, USA), 1.5 \u0026micro;L of DNA template, 1 \u0026micro;L of 10 mM dNTPs, 5 \u0026micro;L of 5\u0026times; Phusion HF Buffer, and 17.3 \u0026micro;L of sterilized double-distilled water. The PCR reactions were performed in a ThermoFisher SensoQuest Labcycler thermal cycler. The PCR program for all genes was 98 ℃ for 30 s, followed by 35 cycles of 98 ℃ for 10 s, the appropriate annealing temperature for 30 s, 72 ℃ for 1 to 1.5 min, and a final 10 min elongation at 72 ℃. The annealing temperature was 54 ℃ for the amplification of ITS, 52 ℃ for LSU, and 55 ℃ for \u003cem\u003eTUB\u003c/em\u003e. The amplified products were sent to Tri-I Biotech Incorporation Company (Taipei, Taiwan) for sequencing, and the sequences for the isolates in this study were deposited in the GenBank database (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFungal isolates used in phylogenetic analysis and their GenBank accession numbers\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e \u003cp\u003eGenBank accession number\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIsolate/strain\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHost/substrate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLocation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eITS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLSU\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eTUB\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eApoharknessia eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 142518\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus\u0026nbsp; pellita\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMalaysia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMG934432\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMN162172\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMG934505\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eApoharknessia eucalyptorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 142519\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus \u0026nbsp;pellita\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMalaysia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKY979752\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eKY979807\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eKY979919\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma alsophilae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC WZ0451*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eAlsophila\u0026nbsp; spinulosa\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810625\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810631\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828586\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma alsophilae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC WZ0452\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eAlsophila\u0026nbsp; spinulosa\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810626\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810632\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828587\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma bambusae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC 1206-4*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBambusoideae \u0026nbsp;sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810629\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810635\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828590\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma bambusae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC 1206-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBambusoideae \u0026nbsp;sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810630\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810636\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828591\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma castaneae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCCmy0162*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCastanea\u0026nbsp; mollissima\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMZ156982\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMZ156985\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMZ220325\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma castaneae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCCmy0523\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCastanea\u0026nbsp; mollissima\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMZ156983\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMZ156986\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMZ220326\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma corymbiae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 132529*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCorymbia\u003c/em\u003e\u0026nbsp; sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAustralia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eJX069861\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eJX069845\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma corymbiicola\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 145052*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCorymbia\u0026nbsp; citriodora\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAustralia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMK047425\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMK047476\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMK047577\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma dipterocarpi\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTBRC 1895*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eDipterocarpus\u0026nbsp; tuberculatus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThailand\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKR994682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eKR994683\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma dipterocarpicola\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMFLUCC 21\u0026ndash;0142*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eDipterocarpus\u003c/em\u003e\u0026nbsp; sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThailand\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOM228844\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOM228842\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOM219638\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma dipterocarpicola\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMFLUCC 21\u0026ndash;0114\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eDipterocarpus\u003c/em\u003e\u0026nbsp; sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThailand\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOM228843\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOM228841\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOM219637\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 124807*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus \u0026nbsp;urophylla\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVenezuela\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973512\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973606\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU973575\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCPC 14161\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus\u0026nbsp; camaldulensis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVietnam\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973510\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973604\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU973573\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCPC 12280\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus\u003c/em\u003e \u0026nbsp;sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eUSA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973507\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973601\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU973570\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCPC 115743\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus \u0026nbsp;globulus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eUruguay\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973509\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973603\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU973572\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma inthanonense\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMFLUCC 23\u0026ndash;0262\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePlant litter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThailand\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOR606510\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOR633320\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOR611920\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma mangiferae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKUMCC 18\u0026ndash;0179*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eMangifera\u003c/em\u003e\u0026nbsp; sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMK084824\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMK084825\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma myracrodruonis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eURM 7799*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eAstronium\u0026nbsp; urundeuva\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBrazil\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMG870421\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMK982151\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMN019566\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma myracrodruonis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eURM 8123\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eAstronium\u0026nbsp; urundeuva\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBrazil\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMK982150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMK982152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMN019567\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma machili\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC BW0233*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eMachilus \u0026nbsp;nanmu\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810627\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810633\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828588\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma machili\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSAUCC BW0221\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eMachilus \u0026nbsp;nanmu\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOP810628\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOP810634\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOP828589\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma jasmini\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMFLUCC 23\u0026ndash;0044\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eJasminum grandiflorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThailand\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOQ786078\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOQ786079\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOQ850148\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma oldii\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 115722\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus\u0026nbsp; camaldulensis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAustralia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973535\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973610\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU993864\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma oldii\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 124808*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus\u0026nbsp; camaldulensis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAustralia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973534\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973609\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU993862\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV14; BCRC FU31382\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233353\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233367\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251137\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV19; BCRC FU31383\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233354\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233368\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251138\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV22; BCRC FU31384\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233355\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251139\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV24; BCRC FU31385\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233356\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233370\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251140\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV25; BCRC FU31386\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233357\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233371\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251141\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV26; BCRC FU31387\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233358\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233372\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251142\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV27; BCRC FU31388*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233359\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233373\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251143\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV36; BCRC FU31389\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233360\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233374\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251144\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV58; BCRC FU31390\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233361\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233375\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251145\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eV64; BCRC FU31391\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTaiwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMT233362\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMT233376\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMT251146\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePseudoplagiostoma variabile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCBS 113067*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus \u0026nbsp;globulus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eUruguay\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGU973536\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGU973611\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGU993863\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003csup\u003ea\u003c/sup\u003e Ex-type strains are marked with \u0026ldquo;*\u0026rdquo;.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003csup\u003eb\u003c/sup\u003eITS = internal transcribed spacer; LSU\u0026thinsp;=\u0026thinsp;large subunit of ribosomal DNA; and TUB\u0026thinsp;=\u0026thinsp;β-tubulin-2.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis\u003c/h2\u003e \u003cp\u003eThe three nuclear gene regions used in phylogenetic analysis were ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e. Sequence data of 27 \u003cem\u003ePseudoplagiostoma\u003c/em\u003e isolates were retrieved from the GenBank database (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Multiple sequence alignment of each gene was conducted using ClustalX v. 2.1 (Larkin et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) and these alignments were concatenated using Sequence Matrix v. 1.7.8 (Vaidya et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The best DNA substitution model for each gene region was predicted by jModeltest 2.1.7 (Posada, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). The DNA substitution models used for ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e were SYM\u0026thinsp;+\u0026thinsp;G, TrNef\u0026thinsp;+\u0026thinsp;I, and HKY\u0026thinsp;+\u0026thinsp;I, respectively. A Bayesian phylogenetic tree was constructed with the software Mr. Bayes v. 3.2.6 (Ronquist et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The analysis was run twice with 1\u0026times;10\u003csup\u003e7\u003c/sup\u003e generations, and samples were taken from the posterior probability trees every 1,000 generations. The first 25% of generations were discarded as burn-in. \u003cem\u003eApoharknessia eucalypti\u003c/em\u003e CBS 142518 and \u003cem\u003eApoharknessia eucalyptorum\u003c/em\u003e CBS 142519 were used as the outgroups. A maximum Likelihood (ML) tree was constructed using IQ tree 2.1.3 (Nguyen et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) with 1,000 bootstrap replicate. The matrices and phylogenetic trees were deposited in the TreeBASE database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://purl.org/phylo/treebase/phylows/study/TB2:S31183\u003c/span\u003e\u003cspan address=\"http://purl.org/phylo/treebase/phylows/study/TB2:S31183\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003ePathogenicity tests\u003c/h2\u003e \u003cp\u003eOne-year-old avocado seedlings of the \u0026lsquo;Choquette,\u0026rsquo; \u0026lsquo;Hall,\u0026rsquo; and \u0026lsquo;Hung Shin Yuan\u0026rsquo; cultivars were used in the pathogenicity tests using the \u003cem\u003ePseudoplagiostoma\u003c/em\u003e isolate V27. For each cultivar, three plants were used in one pathogenicity test, and for each plant, two leaves were inoculated. Conidia were harvested from PDA culture plates after 21 days of incubation at 25 ℃ in the dark. Conidia were diluted with the appropriate amount of sterilized distilled water to obtain a concentration of 2\u0026times;10\u003csup\u003e4\u003c/sup\u003e spores/ml. The spore suspension was sprayed on both the upper and lower sides of young avocado leaves until runoff. The inoculated leaves were sealed in zipper bags with moistened tissue paper to maintain high humidity. Leaves sprayed with sterile distilled water were used as the negative control. The inoculated leaves were removed from the zipper bags 2 days after inoculation and kept in a greenhouse with ambient light and temperature. The pathogenicity test was conducted twice. The first trial was conducted from October 18 to November 1, 2023. The mean temperature during this period was 24.91 ℃, with 8.81 h of sunshine per day on average. The second trial was conducted from November 1 to November 15, 2023. The mean temperature during this period was 23.99 ℃, with 8.83 h of sunshine per day. For the inoculation tests on the three cultivars, leaves of inoculated seedlings were observed 2 weeks after inoculation. To fulfill Koch\u0026rsquo;s postulates, the fungi were recovered from the spots on the leaves and identified based on morphological traits and partial \u003cem\u003eTUB\u003c/em\u003e sequences. For the fruit inoculation assay, \u003cem\u003ePseudoplagiostoma\u003c/em\u003e isolate V27 was incubated on PDA agar at 25 ℃ for 7 days. An agar disc 5 mm of diameter was cut from the edge of the fungal colony and placed on a wound on avocado fruit made using a needle (23G \u0026times; 1.25\u0026Prime;). After inoculation, the avocado fruits were placed in a basin, and the basin wrapped in a plastic bag for 2 days. The inoculated fruits were stored at 25 ℃ in darkness. The inoculated fruits were investigated after 7 days of inoculation.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eField symptoms and fungal isolation\u003c/h2\u003e \u003cp\u003eSixty-three isolates were obtained from avocado leaf spots in different avocado orchards. All 63 isolates exhibited similar colony morphologies viewed on the surface and reverse sides of OA (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, D), MEA (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB, E), and PDA (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC, F) plates. The surface of the colony appeared dark gray, gray to grayish, and olive green (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, B, C), and the colony appeared dark gray to olive green when viewed from the reverse side of the plate (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD, E, F). Ten isolates were chosen arbitrarily to conduct the subsequent phylogenetic analysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePhylogenetic analysis\u003c/h3\u003e\n\u003cp\u003eA Bayesian inference phylogenetic tree based on ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e sequences revealed that the ten \u003cem\u003eP. perseae\u003c/em\u003e isolates (V14, V19, V22, V24, V25, V26, V27, V36, V58, V64) formed an independent clade, which was sister to that of \u003cem\u003ePseudoplagiostoma alsophilae\u003c/em\u003e and \u003cem\u003ePseudoplagiostoma machili\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The clade was supported by a Bayesian posterior probability of 100% and ML bootstrap value of 100%.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eTaxonomy\u003c/h2\u003e \u003cp\u003e \u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e C. J. Wu, J. L. Chen, S. S. Tzean, H. F. Ni, sp. nov.\u003c/p\u003e \u003cp\u003e[Mycobank No:] MB839192 (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eEtymology\u003c/strong\u003e \u003cp\u003eIn reference to the host genus name, \u003cem\u003ePersea\u003c/em\u003e.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eTypification\u003c/strong\u003e \u003cp\u003eBeidou, Changhua, Taiwan (23.853461, 120.544106), isolated from a diseased avocado leaf. Ex-type living culture is BCRC FU31388 (V27) (Holotype TNM F0037701). Several other paratype cultures isolated from different locations and diseased leaf samples were maintained via cryopreservation in the Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan (BCRC FU31382\u0026ndash;BCRC FU31387 and BCRC FU31389\u0026ndash;BCRC FU31391). The GenBank accession numbers of the ten \u003cem\u003eP. perseae\u003c/em\u003e isolates are shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eDescription\u003c/strong\u003e \u003cp\u003eOn leaves, Conidiomata acervular, subepidermal, light brown, 107.39 to 205.67 \u0026micro;m in width, 55.51 to 92.62 \u0026micro;m high (145.21\u0026thinsp;\u0026plusmn;\u0026thinsp;31.41 \u0026times; 75.67\u0026thinsp;\u0026plusmn;\u0026thinsp;11.88 \u0026micro;m, n\u0026thinsp;=\u0026thinsp;15), often with pale yellow droplet exudates containing masses of conidia on the leaf surface. Thin conidiomata wall, consisting of one to two layers of \u003cem\u003etextura angularis\u003c/em\u003e; dehiscence irregular, by rupture of the host epidermal tissue. Conidiophores absent. Conidiogenous cells enteroblastic, annellidic, discrete, indeterminate, ovoid, obclavate or obpyriform, smooth, subhyaline to pale brown, with several percurrent proliferations in the apical part, 8.61\u0026ndash;21.68 \u0026times; 5.86\u0026ndash;14.93 \u0026micro;m (14.24\u0026thinsp;\u0026plusmn;\u0026thinsp;3.52 \u0026times; 9.02\u0026thinsp;\u0026plusmn;\u0026thinsp;2.57 \u0026micro;m, n\u0026thinsp;=\u0026thinsp;25). Conidia 17.66\u0026ndash;23.68 \u0026times; 10.62\u0026ndash;16.48 \u0026micro;m (20.15\u0026thinsp;\u0026plusmn;\u0026thinsp;1.35 \u0026times; 13.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31 \u0026micro;m, n\u0026thinsp;=\u0026thinsp;50), acrogenous, ellipsoidal, mostly aseptate, but rarely with one to three septa, smooth, hyaline to pale yellowish grey or pale brown, with a protruding scar at the base. On leaves, ascomata globose, subglobose or irregular, intra- or subepidermal; wall consisting of two to three layers of dark brown to black \u003cem\u003etextura angularis\u003c/em\u003e. Ostioles beaked lateral, neck internally lateral lined with periphysis. Paraphyses absent. Asci unitunicate, elongated clavate or fusiform with blunt or rounded apex, sessile, 8-spored, hyaline. Ascospores fusiform, one-septate, two-celled unequally, upper cell longer and wider, hyaline, smooth, constricted at the septum, with appendage at both ends, 26.15\u0026ndash;30.22 \u0026times; 12.39\u0026ndash;14.94 \u0026micro;m (28.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 \u0026times; 13.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.071 \u0026micro;m, n\u0026thinsp;=\u0026thinsp;50, on \u003cem\u003eC. equisetifolia\u003c/em\u003e needles).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eCulture characteristics\u003c/strong\u003e \u003cp\u003eOn PDA, colonies reached 4.4 cm in diameter after a week of incubation at 25 ℃ in darkness. Surface floccose to felted, gray to grayish green. Reverse dark gray to olive green. The colony features on OA, MEA, and PDA are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eNotes\u003c/strong\u003e \u003cp\u003eA phylogenetic tree based on ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e gene markers showed that \u003cem\u003eP. perseae\u003c/em\u003e formed a well-supported lineage with a high posterior probability (100% from Bayesian inference) and bootstrap value (100% from ML analysis) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eP. perseae\u003c/em\u003e differed from all other \u003cem\u003ePseudoplagiostoma\u003c/em\u003e spp. in terms of its conidia size (18\u0026ndash;24 \u0026times; 12\u0026ndash;14 \u0026micro;m) and shape except for \u003cem\u003eP. alsophilae\u003c/em\u003e (17\u0026ndash;21 \u0026times; 13\u0026ndash;15 \u0026micro;m), \u003cem\u003eP\u003c/em\u003e. \u003cem\u003emachili\u003c/em\u003e (17.5\u0026ndash;23 \u0026times; 10.5\u0026ndash;13.5 \u0026micro;m), and \u003cem\u003eP. mangiferae\u003c/em\u003e (18\u0026ndash;24 \u0026times; 11\u0026ndash;14 \u0026micro;m). However, the conidiomata of \u003cem\u003eP. alsophilae\u003c/em\u003e, \u003cem\u003eP. machili\u003c/em\u003e, and \u003cem\u003eP. mangiferae\u003c/em\u003e were pycnidial, whereas \u003cem\u003eP. perseae\u003c/em\u003e had acervuli. The colony surfaces of both \u003cem\u003eP. alsophilae\u003c/em\u003e and \u003cem\u003eP. machili\u003c/em\u003e on PDA were gray white to creamy white with irregular margins, and the reverse sides were similar (Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The surface of the \u003cem\u003eP. mangiferae\u003c/em\u003e colony on PDA was pink-white to cream and pale yellowish when viewed from the reverse side (Phookamsak et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). By contrast, most \u003cem\u003eP. perseae\u003c/em\u003e colonies were gray to grayish olive green on the surface and dark gray to olive green when viewed from the reverse side.\u003c/p\u003e \u003cp\u003eThe differences in morphology and phylogenetic relatedness indicated that \u003cem\u003eP. perseae\u003c/em\u003e is a new species. The hosts and characteristics of the conidiomata, conidiogenous cells, and conidia of the 16 currently known \u003cem\u003ePseudoplasgiostoma\u003c/em\u003e species are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAsexual morphological features of \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHost\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eConidiomata type and size (in \u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eConidiogenous cell size (in \u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eConidia size (in \u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eReferences\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. alsophilae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAlsophila\u0026nbsp;spinulosa\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 150\u0026ndash;250\u0026nbsp;\u0026times;\u0026nbsp;200\u0026ndash;300\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8\u0026ndash;13\u0026nbsp;\u0026times;\u0026nbsp;1.5\u0026ndash;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17\u0026ndash;21\u0026nbsp;\u0026times;\u0026nbsp;13\u0026ndash;15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eZhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. bambusae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBambusoideae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 200\u0026ndash;250\u0026nbsp;\u0026times;\u0026nbsp;150\u0026ndash;250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026ndash;13\u0026nbsp;\u0026times;\u0026nbsp;1.5\u0026ndash;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13\u0026ndash;20\u0026nbsp;\u0026times;\u0026nbsp;5.7\u0026ndash;7.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eZhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP.\u0026nbsp;castaneae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCastanea mollissima\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 270\u0026ndash;450 \u0026times; 220 \u0026minus;\u0026thinsp;400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8\u0026ndash;35 \u0026times; 1\u0026ndash;2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9\u0026ndash;13.5 \u0026times; 2\u0026ndash;4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. corymbiae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCorymbia\u003c/em\u003e sp. (\u0026lrm;Myrtaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcervular, up to 300 diameter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10\u0026ndash;20 \u0026times; 4\u0026ndash;7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(14\u0026ndash;)16\u0026ndash;18(\u0026ndash;19) \u0026times; (7\u0026ndash;)8\u0026ndash;9(\u0026ndash;10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCrous et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2012\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. corymbiicola\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCorymbia citriodora\u003c/em\u003e, \u003cem\u003eCorymbia variegata\u003c/em\u003e (\u0026lrm;Myrtaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcervular, up to 300 diameter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15\u0026ndash;30 \u0026times; 3\u0026ndash;5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(15\u0026ndash;)16\u0026ndash;17(\u0026ndash;20) \u0026times; (6\u0026ndash;)7(\u0026ndash;8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCrous et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Crous et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. dipterocarpi\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eDipterocarpus tuberculatus\u003c/em\u003e (Dipterocarpaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18\u0026ndash;25 \u0026times; 2.5\u0026ndash;4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14\u0026ndash;36 \u0026times; 7\u0026ndash;11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSuwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP.\u0026nbsp;dipterocarpicola\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eDipterocarpus\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 63\u0026ndash;153 \u0026times; 113\u0026ndash;288\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.0\u0026ndash;11 \u0026times; 1.0\u0026ndash;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.0\u0026ndash;22 \u0026times; 4.0\u0026ndash;7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. eucalypti\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus camaldulensis\u003c/em\u003e, \u003cem\u003eEucalyptus grandis\u003c/em\u003e, \u003cem\u003eEucalyptus robusta\u003c/em\u003e, \u003cem\u003eEucalyptus urophylla\u003c/em\u003e \u003c/p\u003e \u003cp\u003e( Myrtaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcervular to pycnidial, (170\u0026ndash;)180\u0026ndash;200(\u0026ndash;230) wide and (150\u0026ndash;)170\u0026ndash;190(\u0026ndash;220) high\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(6\u0026ndash;)8\u0026ndash;12(\u0026ndash;15) \u0026times; 2\u0026ndash;4(\u0026ndash;6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(15\u0026ndash;)17\u0026ndash;19(\u0026ndash;23) \u0026times; (6.5\u0026ndash;)7\u0026ndash;8(\u0026ndash;8.5); on MEA, (14\u0026ndash;)16\u0026ndash;19(\u0026ndash;22) \u0026times; (6\u0026ndash;)7\u0026ndash;9(\u0026ndash;11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Fan et al., 2018; Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2016\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. inthanonense\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlant litter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 80\u0026ndash;140 height, 70\u0026ndash;120 diameter (103 \u0026times; 109)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.3\u0026ndash;12.3 \u0026times; 1\u0026ndash;2.7 ( 7 \u0026times; 1.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14\u0026ndash;22.6 \u0026times; 6.4\u0026ndash;10.2 (17.5 \u0026minus;\u0026thinsp;8.5 \u0026micro;m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHittanadurage et al., 2023\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP.\u0026nbsp;machili\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMachilus\u0026nbsp;nanmu\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 150\u0026ndash;200\u0026nbsp;\u0026times;\u0026nbsp;100\u0026ndash;250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7\u0026ndash;16\u0026nbsp;\u0026times;\u0026nbsp;2\u0026ndash;3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.5\u0026ndash;23\u0026nbsp;\u0026times;\u0026nbsp;10.5\u0026ndash;13.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eZhang et a. 2023\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. mangiferae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMangifera\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 70\u0026ndash;140 high and 90\u0026ndash;150 diameter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026ndash;11 \u0026times; 3.2\u0026ndash;12.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e18\u0026ndash;24 \u0026times; 11\u0026ndash;14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePhookamsak et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. myracrodruonis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMyracrodruon urundeuva\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, 250\u0026ndash;290 \u0026times; 236\u0026ndash;245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7\u0026ndash;7.5 \u0026times; 2\u0026ndash;3(\u0026ndash;3.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(10\u0026ndash;)12\u0026ndash;15(\u0026ndash;19) \u0026times; (4\u0026ndash;)5\u0026ndash;6(\u0026ndash;7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eBezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. jasmini\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eJasminum grandiflorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, (135\u0026ndash;)140\u0026ndash;200 high, (145\u0026ndash;)150\u0026ndash;230(\u0026ndash;240) diameter (mean\u0026thinsp;=\u0026thinsp;184 \u0026times; 171)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(6.8\u0026ndash;)7.7\u0026ndash;13.7(\u0026ndash;15.6) \u0026times; 1.6\u0026ndash;2.4(\u0026ndash;3.0)\u003c/p\u003e \u003cp\u003e(mean\u0026thinsp;=\u0026thinsp;10.7 \u0026times; 2.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(11.8\u0026ndash;)14\u0026ndash;22 \u0026times; (5.2\u0026ndash;)6.5\u0026ndash;11 (mean\u0026thinsp;=\u0026thinsp;18.5 \u0026times; 9.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGomdola et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. oldii\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus camaldulensis\u003c/em\u003e ( Myrtaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcervular to pycnidial, (265\u0026ndash;)285\u0026ndash;300(\u0026ndash;330) wide and (200\u0026ndash;)220\u0026ndash;250(\u0026ndash;270) high\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(8.5\u0026ndash;)15\u0026ndash;20(\u0026ndash;26) \u0026times; 2\u0026ndash;3(\u0026ndash;4.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(15\u0026ndash;)17\u0026ndash;20(\u0026ndash;23) \u0026times; (6\u0026ndash;)7\u0026ndash;8(\u0026ndash;9); on MEA, (11\u0026ndash;)14\u0026ndash;17(\u0026ndash;20) \u0026times; (6\u0026ndash;)7\u0026ndash;9(\u0026ndash;11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. perseae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePersea americana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcervular, 107.39\u0026ndash;205.67 wide and 55.51\u0026ndash;92.62 high\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.61\u0026ndash;21.68 \u0026times; 5.86\u0026ndash;14.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.82\u0026ndash;24.15 \u0026times; 11.73\u0026ndash;15.57; on MEA 16.61\u0026ndash;21.73 \u0026times; 12.11\u0026ndash;14.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eThis study\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eP. variabile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEucalyptus globulus\u003c/em\u003e ( Myrtaceae)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePycnidial, (145\u0026ndash;)170\u0026ndash;190(\u0026ndash;245) wide and (130\u0026ndash;)160\u0026ndash;180(\u0026ndash;230) high\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(12\u0026ndash;)15\u0026ndash;20(\u0026ndash;23) \u0026times; 2\u0026ndash;3(\u0026ndash;4.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e(12.5\u0026ndash;)15.5\u0026ndash;17.5(\u0026ndash;23.5) \u0026times; (5.5\u0026ndash;)6.5\u0026ndash;8(\u0026ndash;9); on MEA, (6.5\u0026ndash;)15.5\u0026ndash;17(\u0026ndash;19) \u0026times; (6.5\u0026ndash;)7.5\u0026ndash;9(\u0026ndash;10.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003csup\u003ea\u003c/sup\u003eHost: host Sensu lato, including endophyte hosts.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePathogenicity tests\u003c/h2\u003e \u003cp\u003eIn pathogenicity tests, \u003cem\u003eP. perseae\u003c/em\u003e caused leaf spots on the \u0026lsquo;Choquette,\u0026rsquo; \u0026lsquo;Hall,\u0026rsquo; and \u0026lsquo;Hung Shin Yuan\u0026rsquo; cultivars 2 weeks post-inoculation. Needle-like black spots appeared on leaves (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA-D). Some lesions developed blurry halos (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA, C). The spots also appeared on veins on the reverse sides of leaves (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB). The leaves of the negative control (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE), which was inoculated with sterilized water, had no symptoms. To fulfill Koch\u0026rsquo;s postulates, spots on inoculated leaves were cut, sterilized, and incubated on PDA plates, and \u003cem\u003eP. perseae\u003c/em\u003e were identified according to colony morphology and partial \u003cem\u003eTUB\u003c/em\u003e sequence. \u003cem\u003eP. perseae\u003c/em\u003e were re-isolated from all inoculated leaves, thus fulfilled Koch\u0026rsquo;s postulates. Fruit pathogenicity tests were conducted by placing agar discs bearing \u003cem\u003eP. perseae\u003c/em\u003e mycelium on wounded fruits. No symptoms were visible after one week of observation of avocado fruits (data not shown).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis is the first report characterizing a leaf spot disease on avocado leaves, and the causal agent of this disease, \u003cem\u003eP. perseae\u003c/em\u003e is a new species according to analysis of colony morphology, spore size, sporulation structure, host preference, and phylogenetic analysis with ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e gene markers. In pathogenicity tests, \u003cem\u003eP. perseae\u003c/em\u003e was pathogenic on the leaves of the three avocado cultivars tested.\u003c/p\u003e \u003cp\u003eThe ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e loci are commonly used to distinguish species under the genus \u003cem\u003ePseudoplagiostoma\u003c/em\u003e. For example, Cheewangkoon et al., (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) concatenated ITS and \u003cem\u003eTUB\u003c/em\u003e to distinguish \u003cem\u003eP. eucalypti\u003c/em\u003e, \u003cem\u003eP. oldii\u003c/em\u003e, and \u003cem\u003eP. variable\u003c/em\u003e. Many studies of \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species also included ITS, LSU, and \u003cem\u003eTUB\u003c/em\u003e in their phylogenetic analyses (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Crous et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Gomdola et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Hittanadurage et al., 2023; Mu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Phookamsak et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Suwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Other gene markers, such as \u003cem\u003eTEF\u003c/em\u003e and \u003cem\u003eRPB2\u003c/em\u003e, are also used to resolve \u003cem\u003ePseudoplagiostoma\u003c/em\u003e spp. (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Hittanadurage et al., 2023; Mu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Tang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). However, for \u003cem\u003eP. corymbiae\u003c/em\u003e, \u003cem\u003eP. corymbiicola\u003c/em\u003e, \u003cem\u003eP. dipterocarpi\u003c/em\u003e, and \u003cem\u003eP. diptercarpicola\u003c/em\u003e, no \u003cem\u003eRPB2\u003c/em\u003e sequences have been deposited in GenBank or other DNA sequence databases; thus, using \u003cem\u003eRBP2\u003c/em\u003e to distinguish \u003cem\u003ePseudoplagiostoma\u003c/em\u003e spp. might be problematic.\u003c/p\u003e \u003cp\u003e \u003cem\u003eColletotrichum\u003c/em\u003e spp. might have been mistaken as the causal agent of avocado leaf spot disease in field diagnosis in the past. This is mainly because some \u003cem\u003eColletotrichum\u003c/em\u003e spp. are known pathogens and endophytes in avocados (Giblin et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Shetty et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Use of the appropriate methods and timing are crucial to isolate \u003cem\u003eP. perseae\u003c/em\u003e from symptomatic leaves. In this study, leaves with lesions were kept in a moistened box to induce sporulation of \u003cem\u003eP. perseae\u003c/em\u003e. Using the general fungal isolation method, which involves disinfecting the leaf surface and incubating the leaf discs on a PDA plate, the fungal species regenerated from the disease tissue might be \u003cem\u003eColletotrichum\u003c/em\u003e spp. most of the time, since \u003cem\u003eColletotrichum\u003c/em\u003e spp. grow fast and are common endophytes, pathogens, and saprophytes on avocados. However, the chance of \u003cem\u003eP. perseae\u003c/em\u003e being isolated from leaf tissues increases greatly if isolation is performed at an incipient infection stage, i.e., when needle-like black spots have just appeared on the avocado leaves.\u003c/p\u003e \u003cp\u003eAt least eight species in the \u003cem\u003ePseudoplagiostoma\u003c/em\u003e genus are associated with foliar disease. The pathogenicities of \u003cem\u003eP. eucalypti\u003c/em\u003e and \u003cem\u003eP. mangiferae\u003c/em\u003e have been validated on eucalyptus and mango, respectively (Cheewangkoon et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Sankaran et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Zhou et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), whereas \u003cem\u003eP. corymbiae\u003c/em\u003e and \u003cem\u003eP. corymbiicola\u003c/em\u003e were isolated from leaf spots of the lemon-scented gum tree (\u003cem\u003eCorymbia\u003c/em\u003e spp.) (Crous et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). \u003cem\u003eP. alsophilae\u003c/em\u003e, \u003cem\u003eP. bambusae\u003c/em\u003e, \u003cem\u003eP. castaneae\u003c/em\u003e, \u003cem\u003eP. jasmini\u003c/em\u003e, and \u003cem\u003eP. machili\u003c/em\u003e were also isolated from leaf spots on their preferred host plants, e.g., bamboo or chestnut (Mu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In addition, \u003cem\u003eP. dipterocarpi\u003c/em\u003e and \u003cem\u003eP. myracroduonis\u003c/em\u003e are endophytic, living in the leaves of tree species such as the Indian English gurjun tree (\u003cem\u003eDipterocarpus tuberculatus\u003c/em\u003e) and Urunday tree (\u003cem\u003eMyracrodruon urundeuva\u003c/em\u003e) (Bezerra et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Suwannarach et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). \u003cem\u003ePseudoplagiostoma dipterocarpicola\u003c/em\u003e was isolated from dead twigs and fruits of \u003cem\u003eDipterocarpus\u003c/em\u003e sp. (Keruing). \u003cem\u003eP. inthanonense\u003c/em\u003e was isolated from plant litter and might be a saprophyte (Hittanadurage et al., 2023). In Taiwan, three \u003cem\u003ePseudoplagiostoma\u003c/em\u003e spp. have already been documented and validated to be pathogenic on their respective hosts: \u003cem\u003eP. eucalypti\u003c/em\u003e, which causes leaf spots on swamp mahogany (\u003cem\u003eEucalyptus robusta\u003c/em\u003e) (Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), P. \u003cem\u003emangiferae\u003c/em\u003e, the causal agent of mango leaf blotch (Zhou et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e)d \u003cem\u003eperseae\u003c/em\u003e, which causes avocado leaf spot, as demonstrated in this study.\u003c/p\u003e \u003cp\u003eIn the future, the host range, pathogenicity on different avocado cultivars, etiology, and fungicide sensitivity of \u003cem\u003eP. perseae\u003c/em\u003e will be further studied. \u003cem\u003eIn planta\u003c/em\u003e field assays should also be conducted to confirm the effectiveness of fungicides, which have been tested against \u003cem\u003eP. perseae in vitro\u003c/em\u003e. The information gained from these studies will help avocado growers to control \u003cem\u003eP. perseae\u003c/em\u003e, which causes leaf spot disease of avocado. This will in turn benefit the avocado industry.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eStatements and Declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with Ethical Standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no potential conflicts of interest. This research is not involving Human Participants and/or Animals. Therefore, there is no informed consent needed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBezerra, J. D. P., P\u0026aacute;dua, A. P. S. L., Oliveira, T. G. L., Paiva, L. M., Guarnaccia, V., Fan, X., \u0026amp; Souza-Motta, C. M. (2019). \u003cem\u003ePseudoplagiostoma myracrodruonis\u003c/em\u003e (\u003cem\u003ePseudoplagiostomataceae\u003c/em\u003e, \u003cem\u003eDiaporthales\u003c/em\u003e): a new endophytic species from Brazil. \u003cem\u003eMycological Progress\u003c/em\u003e, 18, 1329\u0026ndash;1339.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCheewangkoon, R., Groenewald, J. Z., Verkley, G. J. M., Hyde, K. D., Winfield, M. J., Gryzenhout, M., Summerell, B. A., Denman, S., Toanun, C., \u0026amp; Crous, P. W. (2010). Re-evaluation of \u003cem\u003eCryptosporiopsis eucalypti\u003c/em\u003e and \u003cem\u003eCryptosporiopsis\u003c/em\u003e-like species occurring on \u003cem\u003eEucalyptus\u003c/em\u003e leaves. \u003cem\u003eFungal Diversity\u003c/em\u003e, \u003cem\u003e44\u003c/em\u003e, 89\u0026ndash;105.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCrous, P. W., Luangsa-Ard, J. J., Wingfield, M. 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United States Department of Agriculture Yearbook.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, Z., Liu, X., Tao, M., Liu, X., Xia, J., Zhang, X., \u0026amp; Meng, Z. (2023). Taxonomy, phylogeny, divergence time estimation, and biogeography of the family \u003cem\u003ePseudoplagiostomataceae\u003c/em\u003e (\u003cem\u003eAscomycota\u003c/em\u003e, \u003cem\u003eDiaporthales\u003c/em\u003e). \u003cem\u003eJournal of Fungi\u003c/em\u003e, 9, 82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou, Z. Y., Tsao, W. C., Chung, W. H., \u0026amp; Wang, C. L. (2022). First report of mango leaf blotch caused by \u003cem\u003ePseudoplagiostoma mangiferae\u003c/em\u003e in Taiwan. \u003cem\u003ePlant Disease\u003c/em\u003e, \u003cem\u003e106\u003c/em\u003e, 2749.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"european-journal-of-plant-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ejpp","sideBox":"Learn more about [European Journal of Plant Pathology](http://link.springer.com/journal/10658)","snPcode":"10658","submissionUrl":"https://www.editorialmanager.com/ejpp/default2.aspx","title":"European Journal of Plant Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Pseudoplagiostoma perseae, avocado, leaf spot disease, pathogenicity test","lastPublishedDoi":"10.21203/rs.3.rs-4019244/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4019244/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLeaf spot disease is commonly found on leaves of avocado plants in avocado orchards in Taiwan. Needle-like spots with yellow halos appear on diseased avocado leaves at the incipient stage, and these spots gradually enlarge and become brown lesions with black-brown centers. The causal agent of this disease is unknown. A \u003cem\u003ePseudoplagiostoma\u003c/em\u003e species was collected from these leaf spots. Evaluation of morphological traits and phylogenetic analysis using the ribosomal DNA internal transcribed spacer operon, β-tubulin, and partial large subunit of ribosomal DNA markers identified this fungal species as a novel species, \u003cem\u003ePseudoplagiostoma perseae\u003c/em\u003e sp. nov. A pathogenicity test was conducted on three avocado cultivars, \u0026lsquo;Choquette,\u0026rsquo; \u0026lsquo;Hall,\u0026rsquo; and \u0026lsquo;Hung Shin Yuan.\u0026rsquo; After 2 weeks of inoculation, needle-like leaf spots appeared on all three cultivars, and \u003cem\u003eP. perseae\u003c/em\u003e was re-isolated from the spots of all inoculated leaves, fulfilling Koch\u0026rsquo;s postulates. This is the first report describing the novel species \u003cem\u003eP. perseae\u003c/em\u003e, which was validated as the causal agent of avocado leaf spot disease.\u003c/p\u003e","manuscriptTitle":"Pseudoplagiostoma perseae sp. nov. causes leaf spot disease on avocado leaves in Taiwan","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-11 19:24:08","doi":"10.21203/rs.3.rs-4019244/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision","date":"2024-05-07T06:44:06+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-03-11T02:36:17+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-03-07T07:52:27+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"European Journal of Plant Pathology","date":"2024-03-07T07:14:33+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-06T23:50:39+00:00","index":"","fulltext":""},{"type":"submitted","content":"European Journal of Plant Pathology","date":"2024-03-06T01:02:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"european-journal-of-plant-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ejpp","sideBox":"Learn more about [European Journal of Plant Pathology](http://link.springer.com/journal/10658)","snPcode":"10658","submissionUrl":"https://www.editorialmanager.com/ejpp/default2.aspx","title":"European Journal of Plant Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"125d9a63-82c0-418c-a99d-d408ef78d8d4","owner":[],"postedDate":"March 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-05T16:08:14+00:00","versionOfRecord":{"articleIdentity":"rs-4019244","link":"https://doi.org/10.1007/s10658-024-02921-1","journal":{"identity":"european-journal-of-plant-pathology","isVorOnly":false,"title":"European Journal of Plant Pathology"},"publishedOn":"2024-08-01 15:57:09","publishedOnDateReadable":"August 1st, 2024"},"versionCreatedAt":"2024-03-11 19:24:08","video":"","vorDoi":"10.1007/s10658-024-02921-1","vorDoiUrl":"https://doi.org/10.1007/s10658-024-02921-1","workflowStages":[]},"version":"v1","identity":"rs-4019244","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4019244","identity":"rs-4019244","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}