Gray Lesions Research Articles

First Report of Paramarasmius palmivorus Causing Collar Rot of Solanum aethiopicum gilo in Ghana

African eggplant (Solanum aethiopicum gilo group) is a nutritious vegetable widely commercialized in Ghana. In the 2021 planting season (May-July), collar rot symptoms were observed on African eggplant on a farm at Domeabra, Legon, and Okumaning in the Central (N5° 48′ 11″, W1° 26′ 48″), Greater Accra (N5° 39′ 34″, W0° 11′ 34″) and Eastern (N6° 8′ 34″, W0° 55′ 59″) regions of Ghana, respectively. Disease incidence was 8-15% in the different farms. Infected plants showed gray-colored lesions at the collar region, soft root rot, and, eventually, wilted due to stem girdling. Thin mycelia were observed on the affected parts of the stem and roots. To identify the causal agent, small portions of diseased stem tissues were submerged in 2% NaOCl for 2 min and rinsed thrice in sterile distilled water (SDW). Next, they were transferred to PDA, amended with ampicillin (50 mg/ml), and then incubated at 25°C for 3 days. Twelve isolates purified by the hyphal-tip culture method were similar morphologically, fluffy with a smooth texture like fine chamois, initially cottony white, and turning pale orange at the center with aging. They were fast-growing, reaching 63.7 to 72.1 cm in diameter. Hyphae were hyaline, septate, branched, 1.4 to 3.1 µm in diameter. These morphological features resembled Paramarasmius sp., classified originally as Marasmius (Sharples) (Antonín et al., 2022). Genomic DNA was extracted from the mycelia of three isolates from the various sampling regions for molecular identification. Two rDNA regions, the internal transcribed spacer (ITS) and mitochondria small subunit (mtSSU), were amplified and sequenced using the ITS1/ITS4 and MS1/MS2 primers, respectively (White et al., 1990). BLASTn analysis of the ITS and the mtSSU sequences revealed a 99.9% and 99.8% identity to P. palmivorus (MN871733.1 and MN794124.1, respectively), further confirmed by phylogenetic analyses. To confirm pathogenicity, the inoculum was made by transferring mycelial plugs from three isolates into autoclaved millet grains, pre-soaked in SDW for 6 hrs. The cultures were incubated at 25 ± 1°C for 8 days, with regular shaking every 2 days to ensure uniform growth. Subsequently, fungal inoculation was made by placing three grains colonized by mycelia close to the stem base of a 21-day-old healthy seedling of African eggplant (cv. Kotobi) raised in pots filled with sterilized potting soil. Each fungal treatment consisted of six replicates of inoculated plants. Three plants were mock-inoculated with autoclaved grains to serve as control treatments, and the experiment was repeated twice. The plants were kept in a screen house with a 12/12 hrs light-dark cycle at 28 ± 3°C. Daily irrigation was done using SDW. Five days after inoculation, typical stem rot symptoms developed at the collar region, characterized by grey water-soaked lesions and thin white mycelium. Next, the seedlings toppled, wilted, and died within 7-14 days. No symptoms were observed in the mock-inoculated plants. The pathogen was reisolated from infected plants on PDA and confirmed as P. palmivorus according to the molecular approach outlined above. P. palmivorus has been recorded to cause bunch rot in oil palm and thread blight disease of cocoa in Ghana (Amoako-Attah et al., 2020). To our knowledge, this study is the first record of P. palmivorus affecting African eggplant in Ghana, and it emphasizes the need to control and determine the host range of this pathogen.

Application of T1-weighted and augmented T1-weighted images of multi-parametric MR sequence in detecting neonatal punctate white matter lesions.

Punctate White Matter Lesion (PWML) is common in neonates. Multi-parametric MR imaging with flexible design (MULTIPLEX, MTP) generates multiple requires only about 6 min for full-head coverage. This study aimed to evaluate the value of T1WI and aT1WI contrasts of MTP in detecting neonatal punctate white matter lesions. Twenty-one neonates with punctate white matter damage underwent multi-parametric MR imaging between November 2022 to July 2024. For subjective image quality, two pediatric neuroradiologists assessed overall image quality, and visualization of structures using a 4-point assessment scale. To analyze objective image quality, the signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), contrast, number and sharpness of lesions were quantified. With regard to sharpness of the lesion, MTP T1WI and aT1WI are comparable to conventional T1W. For subjective assessment, MTP-T1WI exhibited superior overall image quality and anatomical structure display compared to conventional T1WI (P < 0.01). Regarding objective assessment, MTP-T1WI had significantly higher SNR values for gray matter, white matter and lesions than the other two groups. The CNR values of MTP-T1WI and MTP-aT1WI of the white matter to lesion (WM-Lesion) were higher than conventional T1WI. The contrast of aT1WI surpassed that of the other two groups in WM-Lesion contrast. MTP-aT1W can detect more white matter lesions than conventional T1WI (conventional T1WI vs MTP-T1WI vs MTP-aT1WI,123 vs 165 vs 161). The MTP-T1W and aT1W images can enhance lesion contrast and precisely delineate the extent and boundaries of the lesions, and could be more sensitive to PWML than conventional T1WI.

First report of Alternaria leaf spot caused by Alternaria brassicae on broccoli in North Carolina, United States.

In September 2023, broccoli (Brassica oleracea var. italica) 'Sweet Bunch' plants on an organic farm in Buncombe County, North Carolina (NC), displayed symptoms of Alternaria leaf spot. Disease affected 10 to 20% of leaf area on all (approximately 30) plants. Lesions were dark brown with chlorotic halos, irregular in shape, and ≤3 cm in diameter. Leaf samples were incubated in a humidity chamber for 24 h, and the pathogen was putatively identified as Alternaria brassicae based on conidial morphology (Rimmer et al. 2007; Supplemental Fig. 1). Leaf pieces (2 x 2 mm) spanning lesion edges were surface-disinfested (1% sodium hypochlorite solution) for 1 min, rinsed in sterile distilled water, embedded in potato dextrose agar, and incubated at 22°C with a 12-h photoperiod. A pure culture was obtained by transferring a hyphal tip onto 20% V8 juice agar and incubated as above. After 10 d, colonies consisted of concentric zones of white to brown mycelia with white, aerial mycelia. Within 7 d, conidia measuring 80 to 150 µm long with 0 to 3 longitudinal septa, 6 to 15 transverse septa, and a long beak were observed, and the isolate was putatively identified as A. brassicae (Rimmer et al. 2007). To confirm identification, DNA was extracted from mycelia and subjected to PCR using primers to amplify a portion of the internal transcribed spacer of rDNA (ITS: ITS1/ITS4) (White et al. 1990), the transcription elongation factor-1 (tef1: EF1-728F/EF1-986R) (Carbone and Kohn 1999), and the glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH: gpd1/gpd2) (Berbee et al. 1999). Amplicons were sequenced at Eton Bioscience (Durham, NC) and subjected to BLAST analysis in NCBI GenBank. Sequences matched (99.5 to 100% identity) A. brassicae CBS 116528 accessions KC584185 (ITS), KC584641 (tef1), and KC584102 (GAPDH) (Woudenberg et al. 2013). All sequences were deposited in NCBI GenBank (accessions PP584506, PP584507, PP584508). To confirm pathogenicity on broccoli and kale (B. oleracea var. sabellica), 5-week-old plants of broccoli 'Sweet Bunch', 'Eastern Crown', and 'Green Magic' and kale 'Winterbor', 'Darkibor', and 'Oldenbor' grown in 10-cm-diameter pots were separated by crop type and arranged in four randomized complete blocks by cultivar. Each block contained single-plant replicates of each cultivar that were either inoculated or non-inoculated. Conidia of A. brassicae obtained from 10-d-old cultures (3.5 x 103 conidia/ml) were sprayed onto each inoculated plant until run-off. Non-inoculated plants were sprayed with distilled water. Plants were incubated in a humidity chamber alternated with ambient greenhouse conditions every 48 h for 10 d, with average greenhouse temperatures ranging from 15 to 28°C. The experiment was conducted twice. Dark brown to gray lesions measuring 1 to 3 mm in diameter were observed on leaves of all inoculated plants within 5 d of inoculation (Supplemental Fig. 2), and non-inoculated plants remained healthy. On broccoli plants, lesions expanded in concentric rings, and sporulation of the pathogen was observed on leaf tissue and petioles 10 d after inoculation. Lesions remained small (< 3 mm) on kale plants, and sporulation was not observed. Alternaria brassicae was re-isolated from leaf tissue of all inoculated cultivars using methods described above. Pathogen identity was confirmed via colony and conidial morphology. The larger impact of A. brassicae on brassicas is unknown, but finding this less common Alternaria species in the eastern United States could have implications for host resistance development and fungicide efficacy (Nieto-Lopez et al. 2023).

A Mycovirus Representing a Novel Lineage and a Mitovirus of Botrytis cinerea Co-Infect a Basidiomycetous Fungus, Schizophyllum commune.

Strain IBc-114 was isolated from a gray mold lesion and was identified as the fungus Schizophyllum commune. In this strain, two mycoviruses, Schizophyllum commune RNA virus 1 (ScRV1, C_AA053475.1) and Botrytis cinerea mitovirus 9 strain IBc-114 (BcMV9/IBc-114, C_AA053476.1), were isolated and characterized. ScRV1 has flexuous filamentous particles about 20 ± 2.1 nm in diameter and 1000 ± 94.2 nm in length. The genome of ScRV1 is 7370 nt in length and contains two open reading frames (ORFs) which encode a polyprotein and a coat protein, respectively. The polyprotein has 1967 aa, including a helicase domain and an RdRp domain which has the highest identity of 28.21% with that of Entomophthora benyvirus E (EbVE). The coat protein has 241 aa which is mostly phylogenetically close to the coat proteins of Alphatetraviridae. Based on the phylogenetic analysis of ScRV1 and viruses selected, ScRV1 might represent a new family (temporarily named Mycobenyviridae) of the order Hepelivirales. The genome of BcMV9/IBc-114 that infects S. commune is 2729 nt in length and has only one ORF encoding an RdRp protein with 719 aa. BcMV9/IBc-114 has the highest identity of 98.61% with Botrytis cinerea mitovirus 9 (BcMV9) (MT089704). ScRV1, but not BcMV9/IBc-114, has certain effects on the host growth of S. commune. Furthermore, BcMV9/IBc-114 has been demonstrated to replicate in the ascomycetous fungi Botrytis cinerea and Sclerotinia sclerotiorum, and it negatively affects the growth and pathogenicity of B. cinerea, but it does not affect S. sclerotiorum. This is the first report of mycoviruses in S. commune and cross-phyla transmission of mitovirus in nature.

First report of Exserohilum turcicum causing leaf blight on sudangrass (Sorghum × drummondii) in Mexico

In February 2021, elongated grey lesions with reddish margin were observed on sudangrass (Sorghum × drummondii) leaves, in the municipality of Guasave, Sinaloa, Mexico. These symptoms resembled those of leaf blight caused by Bipolaris and Exserohilum on other grasses. Two fungal isolates were recovered from symptomatic leaves, and their identities were confirmed based on a polyphasic approach. The morphological and phylogenetic analysis of the actin gene (act) and internal transcribed spacer region (ITS) of both isolates revealed their identity as Exserohilum turcicum. A pathogenicity test with a representative isolate confirmed this fungus as the causal agent of leaf blight on sudangrass, as symptoms previously observed in the field were developed on the inoculated plants. The E. turcicum isolate was recovered from the diseased sudangrass plants, fulfilling Koch’s postulates. This is the first report of E. turcicum causing leaf blight on sudangrass in Mexico.

First Report of Grovesinia moricola Causing Zonate Leaf Spot on Hernandia nymphaeifolia in Taiwan.

Hernandia nymphaeifolia (C. Presl) Kubitzki, a native tree of Taiwan, is a sea drift plant (Yang and Lu 1996). It is a salt- and wind-tolerant tree (Bezona et al. 2009) and was selected for the afforestation of badlands in coastal areas of Taiwan. In December 2022, all H. nymphaeifolia seedlings at a nursery in Wu-Lai, Taiwan were diseased and wilted with a similar progression. The initial symptom was small zonate white or gray lesions with water-soaked periphery on leaves. Then, expansion and fusion of leaf spots which caused leaf blight and defoliation were observed. Seedlings eventually wilted. Sporophores found on the host were generally hypophyllous, solitary, erect, and easily detachable. The upper portion of the sporophore was considered an individual conidium and consisted of a pyramidal head that was fusiform to ventricose, 206.3 to 501.8 μm (average: 378.0 ± 75.3 μm) long, and 63.6 to 104.5 μm (average: 85.0 ± 16.2 μm) wide at the broadest point (n=30). Branches within the pyramidal head were short, compact, and di- or trichotomously branched. The central stipe was hyaline, broad, septate, tapering toward an acute apex, and sometimes constricted at the basal septum. Sclerotia were gray or black, spherical, and 1.0 to 2.5 mm (n=10) in diameter and observed on older lesions. The fungus was isolated from infected tissue and sporophores and maintained on potato dextrose agar (PDA) at 20°C in darkness. Sclerotia were produced on PDA after 4 to 5 weeks and were irregular or spherical, but no sporophore was developed. The fungus was identified as Grovesinia moricola (I. Hino) Redhead based on morphological characteristics (Tomoko et al. 2006). Three DNA samples was obtained from the cultures isolated from the diseased leaf, sporophores and sclerotia. They were then amplified by PCR with primers for the internal transcribed spacer region (ITS; primers ITS5/ITS4) and the large subunit nuclear ribosomal RNA gene (LSU; primers LR0R/LR5) (Cho et al. 2017), and then sequenced respectively. The sequences were deposited into GenBank with accession nos. PP727191 to PP727193 and PP748518 to PP748520. BLAST analysis of the three isolates showed 100% identity to the sequences of G. moricola from Taiwan (OP550202, OP550203) for the ITS region and 99.9% identity to the sequence of G. moricola from the USA (MW013804) for the LSU rRNA gene. The specimens (FS2022-140) and the culture (Asco-0109) in this study were deposited into the herbarium of Taiwan Forestry Research Institute in Taiwan. Koch's postulates were performed by inoculating four 8-month-old, asymptomatic, potted H. nymphaeifolia plants; every plant was inoculated with sporophores from infected leaves on the upper surface of each of five leaves. Four uninoculated plants were kept in separate pots and served as controls. All plants were covered with transparent plastic bags individually and incubated in a growth chamber at 18 to 20°C with 8 h of light. Similar leaf spots and sporophores were observed after 2 to 4 days and 10 days on every inoculated plant but not on uninoculated plants. The pathogen with a similar colony on PDA was reisolated from the leaf spots of the inoculated plants. Molecular identification of the reisolated pathogen by the above method was carried out. The sequences showed 99.9% identity to the sequence of G. moricola, and were deposited into GenBank with accession nos. PQ157896 to PQ157897 (ITS region) and PQ157701 to PQ157702 (LSU rRNA gene). The pathogenicity test was repeated once. G. moricola is known to cause severe defoliation on woody and annual plants, including at least 73 host species and 36 families distributed in the eastern United States and Japan (Trolinger et al. 1978). This is the first report of G. moricola on H. nymphaeifolia in the world. Control of the disease would play an important role in maintaining healthy seedlings for the afforestation in Taiwan.

Epidemiological and Clinico-therapeutic Studies on Dermatophytosis in Cattle at Parbhani

Background: To investigate prevalence, risk factors, diagnosis and alternative treatment regimens of dermatophytosis in cattle in and around Parbhani. Methods: The overall period of prevalence study was from January, 2022 to December, 2022 in and around Parbhani. The presumptive diagnosis was done by SDA culture media and treatment was given to 18 selected positive cattle which was equally categorized into three group viz; Clove oil, Cow urine distillate and whitfield ointment/control group. Result: The overall prevalence recorded was 17.2 per cent. The most affected cattle were 4 to 8 years of age group, non-descript breed and male cattle. Dermatophytosis in adult cattle is typically manifested as a circumscribed, crusty, whitish grey lesion with focal alopecia, whereas in calf asbestos, crusty lesion was observed. The hump, neck and back were the major sites of occurrence for dermatophytosis. On cultural examination, Trichophyton verrucosumwas mostly isolated. On haematological investigation granulocytosis, monocytosis and lymphopenia were observed in affected cattle. The recovery period for Clove oil was 7 days, Cow urine distillate showed recovery in days and cattle treated with whitfield ointment recovered in 19 days.

Colletotrichum aeschynomenes Causing Anthracnose in Combretum indicum in Guangxi, China.

Combretum indicum（L.）Jongkind, distributed in Southeast Asia, is widely planted in southern China for its ornamental and medicinal value. In February 2023, anthracnose symptoms were observed on C. indicum leaves in Nanning Garden Expo (N22°43', E108°28'), Guangxi, China, causing severe defoliation of infected plants with a foliar disease incidence ranging from 40 to 60% (n = 100) in a 2 ha field. Disease symptoms began with small red spots (2 to 3 mm by 2 to 3 mm) on the leaves and gradually enlarged to larger irregular light grey lesions with yellowish halos (3 to 5 mm by 2 to 8 mm). In the late stage, spots merged into larger irregular lesions (5 to 15 mm by 6 to 13 mm) and the necrotic lesions abscised. Three diseased samples in total were collected from plants in three different locations. Symptomatic leaves were cut into small pieces (3×3 mm), disinfected with 75% ethanol solution for 10 s, 2% NaClO for 1 min followed by three washes in sterile distilled water. Tissue pieces were separately plated on potato dextrose agar (PDA) and incubated at 25°C for five days. One representative isolate from each sample (SJ-1, SJ2-1 and SJ3-1) were chosen for further analysis. Colonies were villiform, initially white, later turning gray in 7 days on PDA at 25℃. The average diameter for colonies were 8.1 cm, 8.0 cm and 8.1 cm for SJ1-1, SJ2-1 and SJ3-1, respectively. Conidia were aseptate, hyaline, cylindroid, and averaged 11.94 μm × 5.04 μm, 11.78 μm × 5.14 μm and 11.74 μm × 4.59 μm (n=90) for SJ1-1, SJ2-1 and SJ3-1, respectively. The characteristics were close to the descriptions of Colletotrichum spp. (Weir et al. 2012). Genomic DNA was extracted from 7-day-old aerial mycelia of these isolates. The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), β-tubulin (TUB2), chitin synthase (CHS-1), calmodulin (CAL) and the intergenic region between apn2 and MAT1-2-1 (ApMat) were amplified using primers ITS1/ITS4 (White et al. 1990), GDF/GDR, ACT-512F/ACT-783R, T1/Bt2b, CHS-79F/CHS-354R, CL1C/CL2C (Weir et al. 2012) and AM-F/AM-R (Silva et al. 2012), respectively. Sequences were deposited in GenBank (ITS: OR540240-OR540242; GAPDH: PP328968-PP328970; ACT: PP328959-PP328961; TUB2: PP328971-PP328973; CHS-1: PP328965-PP328967; CAL: PP328962-PP328964 and ApMat: OR548253-OR548255). A phylogenetic analysis was made via Bayesian inference based on the concatenated sequences (ITS, GAPDH, ACT, TUB2, CHS-1, CAL and ApMat). According to morphology and phylogenetic analysis, SJ1-1, SJ2-1 and SJ3-1 were identified as C. aeschynomenes. Pathogenicity was confirmed on leaves with and without wounds of 24 one-year-old C. indicum plants in a greenhouse in Nanning, Guangxi Province. The wound was made with a sterilized needle. Wounded and unwounded leaves were inoculated with 20 μl of conidial suspension (106 spores/ml in 0.1% sterile Tween 20) of the three isolates and control plants were inoculated with water containing 0.1% sterile Tween 20 (6 leaves/plant, 3 plants/treatment). All plants were covered with plastic bags to maintain a high humidity environment and placed in a 28°C growth chamber with constant light. After 7 days of incubation, necrotic lesions were observed on inoculated wounded leaves, whereas unwounded leaves and control plants showed no symptoms. The fungi were re-isolated from symptomatic leaves, completing Koch's postulates. These species can cause severe diseases in a variety of plants worldwide, such as Manihot esculenta, Theobroma cacao and Myrciaria dubia (Sangpueak et al. 2018; Nascimento et al. 2019; Matos et al. 2020). To our knowledge, this is the first report of C. aeschynomenes causing C. indicum leaf anthracnose in China. The results will provide valuable information for management of anthracnose in C. indicum.

First report of leaf spot on Daphniphyllum macropodum caused by Neopestalotiopsis clavispora in China.

Daphniphyllum macropodum Miq., an evergreen arbor, is widely cultivated in southern China for its ornamental and medicinal value (Su et al. 2013). In October 2019, a severe leaf spot was observed on D. macropodum in Jinggangshan National Nature Reserve in Ji'an city, Jiangxi, China (114°06'23″E, 26°32'25″N). The plants were about 15 years old, and the disease incidence was estimated to be 15% (4/26 plants). The disease primarily appeared as small black spots on the leaves. At the late stage, the spots enlarged and coalesced into regular or irregular gray necrotic lesions with dark margins. We collected five samples per plant and total 20 samples were collected to isolated the pathogen strains. The margin of the diseased tissues was cut into 5 mm × 5 mm pieces; surface disinfected with 70% ethanol and 2% NaOCl for 30 s and 60 s, respectively; and rinsed thrice with sterile water. Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C in the dark. Pure cultures were obtained by single-spore isolation method, and the representative isolates, JRM3, JRM6, and JRM8 were used for morphological studies and phylogenetic analyses. The colonies of three isolates grown on PDA were white, cottony, and flocculent, contained undulate edges with dense aerial mycelium on the surface at 25 °C. Conidiomata was black conidial masses on PDA. Conidia were 5-celled, clavate to fusiform, smooth, 19.3 to 24.4 long × 6.1 to 8.6 μm wide (n = 50). The 3 median cells were dark brown to olivaceous, the central cell was darker than the other 2 cells, and the basal and apical cells were hyaline. All conidia developed one basal appendage (3.4 to 8.3 μm long; n = 50), and 2 to 3 apical appendages (18 to 32 μm long; n = 50), filiform. The morphological characteristics of the isolates are comparable with those of the genus Neopestalotiopsis (Maharachchikumbura et al. 2014). The internal transcribed spacer (ITS) regions, β-tubulin 2 (TUB2) and translation elongation factor 1-alpha (TEF1-α) were amplified from genomic DNA for the three isolates using primers ITS1/ITS4, T1/Bt-2b, EF1-728F/EF-2 (Maharachchikumbura et al. 2014), respectively. The sequences of the isolates were submitted to GenBank (ITS, OQ372202 to OQ372204; TUB2, OQ390129 to OQ390131; TEF1-α, OQ390126 to OQ390128). A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences placed JRM3, JRM6, and JRM8 in the clade of N. clavispora. Based on the multi-locus phylogeny and morphology, three isolates were identified as N. clavispora. To confirm pathogenicity, eight healthy 10-year-old D. macropodum plants growing in the field were chosen, and 4 leaves per plant were wounded with a sterile needle and inoculated with 10 μL conidial suspension per leaf (106 conidia/ml). Eight plants inoculated with sterile water were used as control. All the inoculated leaves were covered with plastic bags to maintian a humidity environment for 2 days. The leaves inoculated with conidial suspension showed similar symptoms to those observed in the field, whereas control leaves were asymptomatic for 10 days. The same fungus were re-isolated from the lesions, whereas no fungus was isolated from control leaves. N. clavispora was determined as the pathogen of a variety of plant diseases, including Kadsura coccinea (Xie et al. 2018), Dendrobium officinale (Cao et al. 2022), Macadamia integrifolia (Santos et al. 2019). However, this is the first report of N. clavispora infecting D. macropodum in China. This work provided crucial information for epidemiologic studies and appropriate control strategies for this newly emerging disease.

First report of Pseudomonas cichorii causing bacterial leaf blight of pocketbook plant (Calceolaria hybrida) in Taiwan.

Pocketbook plants (Calceolaria spp.) are flowering ornamentals often grown as potted plants (Poesch 1937). In December 2022, leaf blight symptoms were observed on 2-mo-old plants of C. hybrida F1 'Dainty'. The disease was found in a nursery in Ren'ai Township, Nantou, and about 20% of the plants exhibited symptoms. Symptomatic plants had brown or gray necrotic lesions of different sizes and shapes, mostly around leaf margins. Lower leaf wilting was also observed (Fig. S1, A and B). Three plants were sampled. Leaf lesions were surface-disinfected with 75% ethanol and cut into smaller pieces in 10 mM MgCl2. After observing bacterial streaming under a microscope, the bacteria were streaked onto nutrient agar (NA). Following 2 days at 28°C, a type of round, creamy white colony predominated on all the plates. Three strains (Calc-A, Calc-B, and Calc-C) were obtained, one from each plant. The strains produced fluorescent pigments on King's B medium and were tested Gram-negative. The strains were characterized with the LOPAT scheme (Schaad et al. 2001). They did not exhibit activities of pectic enzymes, arginine dihydrolase and levan sucrase, but produced oxidase and induced the hypersensitive response in tobacco. DNA was extracted from the strains for PCR amplification of the 16S rDNA with primer pair 27f/1492r as described by Lane (1991). The 16S rDNA sequences were compared with entries in the GenBank database. The sequences obtained (GenBank accession no. OR824302) matched that of Pseudomonas cichorii MAFF 301158 (accession no. AB724288; 1,403/1,403 bp) and were 99% identical to that of DSM 50259T (accession no. CP074349; 1,391/1,405 bp). The strains were also tested with the species-specific primers hrp1a and hrp2a (Cottyn et al. 2011). The amplicons were sequenced and a BLASTn search showed that the sequences (accession no. OR827305) shared the highest identity (99.3%) with that of P. cichorii strain 83-1 (accession no. DQ168848; 848/854 bp) and were 97.3% identical to the sequence of DSM 50259T (accession no. CP074349; 831/854 bp). Calc-A was selected as a representative strain and deposited in the Bioresource Collection and Research Center, Taiwan (reference no. BCRC 81432). Koch's postulates were fulfilled by spray-inoculating a suspension of Calc-A on three 2-mo-old C. hybrida F1 'Dainty' plants. The inoculum was prepared by suspending NA-grown cells in 10 mM MgCl2 including 0.02% Silwet L-77 (OD600 = 0.3; 1.5 x 108 CFU/ml). For the controls, three plants were sprayed with bacteria-free solution. The plants were bagged throughout the experiment and kept in a growth chamber (14/10 h light/dark; 26/24°C day/night). Leaf blight and wilting symptoms developed on all leaves of the inoculated plants after 30 h, but not the controls (Fig. S1, C and D). The pathogen was reisolated from the treatment group, and colony PCR with hrp1a/hrp2a showed that the reisolated strain shared the same sequence with Calc-A to Calc-C. Repeating the inoculation assay produced consistent results. This is the first report of P. cichorii affecting Calceolaria in Taiwan. The bacterium has been reported infecting diverse crops in Taiwan, such as tomato and lettuce (Tsai et al. 2014). Expanding the understanding of the pathogen's potential hosts could help prevent its spread across important crops.

Structural MRI measures are associated with fatigue severity and persistence in a large, real-world cohort of people with multiple sclerosis

Background: Results of research on radiological hallmarks of multiple sclerosis (MS) fatigue have been conflicting. Objective: To investigate the associations of lesion and brain compartment volumes with fatigue severity and persistence in people with multiple sclerosis (PwMS). Methods: The Multiple Sclerosis Partners Advancing Technology and Health Solutions (MS PATHS) network collects standardized data during routine care of PwMS from 10 healthcare institutions. Magnetic resonance imaging (MRI) predictors included baseline brain parenchymal (BPF) and gray matter fractions (GMF) and T2 lesion volume (T2LV). The Quality of Life in Neurological Disorders (Neuro-QOL) fatigue subscore was analyzed linearly and categorically using T-score cutpoints, with a period of elevated symptoms defined as T-score ⩾ mean + 0.5 SD over follow-up. Results: At baseline, of 4012 participants (average age: 45.6 ± 11.8 years; 73% female; 31% progressive MS), 2058 (51%) had no fatigue, 629 (16%) had mild fatigue, and 1325 (33%) had moderate-to-severe fatigue. One SD greater baseline BPF and GMF were associated with 0.83 (p < 0.001) and 0.38 (p = 0.02) lower values in the baseline Neuro-QOL fatigue T-score. A 1 SD lower log of total T2LV was associated with a 0.49 (p < 0.001) lower baseline fatigue T-score. Higher BPF and lower T2LV at baseline were associated with lower odds of subsequent periods of elevated fatigue. Conclusion: Baseline lesion burden and lower generalized whole-brain volumes were associated with MS fatigue in cross-sectional and longitudinal analyses in a large, real-world cohort of PwMS.

Antifungal potential of lipopeptides produced by the Bacillus siamensis Sh420 strain against Fusarium graminearum.

Biological control is a more sustainable and environmentally friendly alternative to chemical fungicides for controlling Fusarium spp. infestations. In this work, Bacillus siamensis Sh420 isolated from wheat rhizosphere showed a high antifungal activity against Fusarium graminearum as a secure substitute for fungicides. Sh420 was identified as B. siamensis using phenotypic evaluation and 16S rDNA gene sequence analysis. An in vitro antagonistic study showed that Sh420's lipopeptide (LP) extract exhibited strong antifungal properties and effectively combated F. graminearum. Meanwhile, lipopeptides have the ability to decrease ergosterol content, which has an impact on the overall structure and stability of the plasma membrane. The PCR-based screening revealed the presence of antifungal LP biosynthetic genes in this strain's genomic DNA. In the crude LP extract of Sh420, we were able to discover several LPs such as bacillomycin, iturins, fengycin, and surfactins using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Microscopic investigations (fluorescent/transmission electron microscopy) revealed deformities and alterations in the morphology of the phytopathogen upon interaction with LPs. Sh420 LPs have been shown in grape tests to be effective against F. graminearum infection and to stimulate antioxidant activity in fruits by avoiding rust and gray lesions. The overall findings of this study highlight the potential of Sh420 lipopeptides as an effective biological control agent against F. graminearum infestations.IMPORTANCEThis study addresses the potential of lipopeptide (LP) extracts obtained from the strain identified as Bacillus siamensis Sh420. This Sh420 isolate acts as a crucial player in providing a sustainable and environmentally friendly alternative to chemical fungicides for suppressing Fusarium graminearum phytopathogen. Moreover, these LPs can reduce ergosterol content in the phytopathogen influencing the overall structure and stability of its plasma membrane. PCR screening provided confirmation regarding the existence of genes responsible for biosynthesizing antifungal LPs in the genomic DNA of Sh420. Several antibiotic lipopeptide compounds were identified from this bacterial crude extract using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Microscopic investigations revealed deformities and alterations in the morphology of F. graminearum upon interaction with LPs. Furthermore, studies on fruit demonstrated the efficacy of Sh420 LPs in mitigating F. graminearum infection and stimulating antioxidant activity in fruits, preventing rust and gray lesions.

First report of Diaporthe eres causing leaf spot on Rhododendron latoucheae Franch. in China.

Rhododendron latoucheae Franch. is an evergreen shrub with charming fragrance and large and abundant flowers that make it highly attractive as an ornamental species. The species is distribution in southwest China covers several different habitats and environments (Zhang, et al. 2022). From May to July in 2023, symptoms of leaf spot were observed on R. latoucheae over a wide portion of the Baili Azalea Forest Area (27°10' to 27°20'N, 105°04' to 106°04'E), Guizhou Province, China. About 500 plants were surveyed, and the incidence of leaf spot on R. latoucheae leaves was 12%, significantly reducing their ornamental and economic value. The affected leaves had irregular, grey white lesions with a clear blackish brown boundary and faint black conidiomata in a brown center. To isolate the pathogen, 15 symptomatic leaves were collected from 10 plants. A few black dots were picked from the lesions with a sterilized needle, plated on water agar, and incubated at 25°C for 24 h to observe spore germination (Choi et al. 1999). Then the germinated spores were transferred onto PDA for further purification and morphological observation. Three single-spore isolates (GULJ1-L7, GULJ1-L8, and GULJ1-L9) identical in morphology were obtained. The isolate GULJ 1-L7 was used for further study. Colonies on PDA irregular grew white felty aerial mycelium, becoming white felted aerial mycelium in the centre and grey-brown mycelium at the marginal area on the upper surface, while the lower surface presents alternating white, tan and taupe. Colony with conidiomata irregularly distributed over agar surface. In the representative isolate, darkly pigmented pycnidia (flask-shaped) were produced over the colony surface on PDA after about 30 days, and oozed milky or yellowish mucilaginous drops. The fungus produced two types of conidia, α and β. Regular α conidia were 5.15-10.29 × 1.54-3.33 μm (n = 50), hyaline, elongated, biguttulate and non-septate. Beta conidia were 20.34-31.91 × 1.01-1.90 μm (n = 50), aseptate, hyaline, smooth, spindle shaped, slightly curved to bent. The morphological features were consistent with the description of Diaporthe eres (Pereira, et al. 2022). The pathogen was confirmed to be D. eres by amplification and sequencing of the internal transcribed spacer region (ITS), the partial β-tubulin (TUB), the partial translation elongation factor 1-alpha (TEF) genes and the calmodulin (CAL) using primers ITS1/ITS4, Bt-2a/Bt-2b, EF1-728F/EF1-986R, and CAL-228F/CAL-737R, respectively (Tao et al. 2020). Sequences from PCR amplification were deposited in GenBank with accession numbers OR740563 (ITS), OR754301 (TUB), OR754298 (TEF), and OR754295 (CAL) respectively. BLAST searches of the sequences revealed 99.07% (533/538 nt), 100% (490/490 nt), 99.69% (317/318 nt) and 98.95% (376/380 nt) homology with those of D. eres AR5193T from GenBank (KJ210529.1, KJ420799.1, KJ210550.1 and KJ434999.1), respectively. Phylogenetic analysis (MEGA 7.0) using the maximum-likelihood method placed the isolate GULJ1-L7 in a well-supported cluster with D. eres CBS 138694T. The pathogen was thus identified as D. eres based on the morphological characterization and molecular analyses (Feng, et al. 2013; Tao, et al. 2020). The pathogenicity of GULJ1-L7 was tested through a pot assay. Due to the difficulty of artificial planting wild R. latoucheae, we conducted a pot essay to detect the pathogenicity of GULJ1-L7 using a closely related Rhododendron delavayi Franch. Ten healthy R. delavayi plants were scratched with a sterilized needle (0.45 mm in diameter) on three leaves per plant. Plants were inoculated by spraying α and β spore mixture suspension (106 spores ml-1) of GULJ1-L7 onto leaves until runoff, and the control leaves were sprayed with sterile water. The plants were maintained at 25°C and 75% relative humidity in a growth chamber. The pathogenicity test was repeated three times. After 14 days, the treated leaves developed brown lesions similar to those in the field, whereas the control had no symptoms. The same fungus was reisolated from the infected leaves and identified based on a morphological characterization and molecular analyses. These results fulfilled Koch's postulates. To our knowledge, this is the first report of leaf spot on R. latoucheae caused by D. eres in China. The fungal pathogen identification will provide valuable information for prevention and management of leaf spot disease associated with R. latoucheae.

Clinicomorphological Analysis of Urinary Bladder Lesions with Special Reference to Immunohistochemical Analysis of Grey Zone Lesions using Cytokeratin 20: A Cross-sectional Study

Clinicomorphological Analysis of Urinary Bladder Lesions with Special Reference to Immunohistochemical Analysis of Grey Zone Lesions using Cytokeratin 20: A Cross-sectional Study

First Report of Exserohilum rostratum Causing Leaf Blight on Broccoli (Brassica oleracea var. italica) in China.

Broccoli (Brassica oleracea var. italica) is not only an important crop worldwide with a large amount of production and consumption annually, but also rich in biologically active compounds (Surh et al., 2021). In November 2022, an unknown leaf blight was observed in the Broccoli planting area, Wenzhou City of Zhejiang Province (28.05 °N, 120.31 °E). Symptoms initially occurred at the leaf margin with yellow to gray lesions that were irregular and wilting. Approximately 10% of the surveyed plants were affected. To determine the pathogen, leaves with blight were collected randomly from five B. oleracea plants. Tissue blocks (3×3 mm) from diseased leaf portions were disinfected with 75% ethanol, rinsed three times with sterilized water, placed aseptically onto potato dextrose agar (PDA) medium, and incubated for 5 days at 28℃ in darkness. Seven fungal isolates with the same morphology were obtained using the spore method. The observed colonies were circular, taupe, pewter in color with light gray edging and many cottony aerial mycelia. Conidia were straight, curved or slightly bent, ellipsoidal to fusiform, and septate (typically 4-8 septa per conidium), with the size of 50.0-90.0 μm × 10.0-20.0 μm (n=30). The conidia had a slightly protruding and truncate hilum. These morphological characteristics were consistent with Exserohilum rostratum (Sharma et al., 2014). To further identify the pathogen, isolate WZU-XLH1 was chosen as a representative and the internal transcribed spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase-like (GAPDH) gene were amplified and sequenced using primer pairs ITS1/ITS4 (White et al., 1990) and Gpd1/Gpd2 (Berbee et al., 1999), respectively. The ITS and gpd gene sequences of isolate WZU-XLH1 were deposited in the GenBank database with accession numbers OQ750113 and OQ714500, respectively. BLASTn analysis showed matches of 568/571 (MH859108) and 547/547 (LT882549) with Exserohilum rostratum CBS 188.68. A neighbor-joining phylogenetic tree was constructed by combining the two sequenced loci, this isolate in the E. rostratum species complex clade at 71% bootstrap support.To verify the pathogenicity of the isolate, ten healthy Broccoli (cultivar 'You Xiu') seedlings with at least five leaves were divided into two groups: one group was inoculated with the isolate, while the other group served as a control. After surface disinfection with 75% ethanol and wiping with sterile water, tiny wounds were made on two leaves (two wounds in one leaf) using an inoculation needle. Fungal culture plugs cut from the isolate were placed on the wounds, while sterile PDA plugs served as the control. The leaves were sealed in wet airtight bags to retain moisture at room temperature with natural light (Cao et al., 2022). After five days, all leaves inoculated with isolate WZU-XLH1 showed symptoms identical to those observed in the field, with no symptoms present in the control group. The pathogenicity was confirmed by repeating the test in triplicate, and fungi re-isolated from symptomatic leaves were identified as E. rostratum by the morphological and molecular methods described above. To the best of our knowledge, this is the first report of E. rostratum causing leaf blight on broccoli in China. This study contributes to our understanding of B. oleracea leaf blight and establishes a basis for future studies on E. rostratum to develop management strategies.

First report of Neopestalotiopsis clavispora causing leaf blight on Aegicerascorniculatum in China.

Aegicerascorniculatum is an important mangrove plant that mainly grows in tropical and subtropical regions. Ithasimportantecological,economicandsocialbenefits (Bandaranayake 1998). In April 2021, a leaf disease on A.corniculatum was observed in Zhanjiang (21.21° N, 110.41° E), Guangdong province, China. Diseaseincidenceandseverity were 15%and 20% (n = 100 investigated plants), respectively. The disease symptoms on leaves primarily appeared as small brown spots, then enlarged and coalesced into regular or irregular gray necrotic lesions with dark margins. At the late stage of symptom development, black acervuli appeared on the necrotic lesions. Ten symptomatic leaves from 10 plants were collected. Small pieces of tissue (4 × 4 mm) were cut from lesion borders and were surfaced disinfected in 75% ethanol for 30 s, followed by 1 min in 1% NaClO, rinsed three times with sterile water, plated on potato dextrose agar (PDA), and incubated at 28°C. After 7 days, a total of 10 fungal isolates with 100% isolation frequency were obtained and three representative strains (THS-1, THS-2, and THS-3) were used for morphological and molecular characterization.Colonies were white with cottony aerial mycelium and irregular margins. Black viscous acervuli were scattered on the colony surface 10 days after incubation. Conidia were spindle shaped, five cells, four septa, 18.77-28.70 × 4.53-6.80μm (mean 23.13 × 5.14μm) in size (n = 50). Basal and apical cells were colorless while the three medium cells were dark brown and lightly versicolor. All conidia had one basal appendage of 3.51-7.27μm (mean 5.49 μm; n = 50) and two to three apical appendages of 15.80-33.64μm (mean 25.87 μm; n = 50). These morphological characteristics are consistent with those of Neopestalotiopsis clavispora(Maharachchikumbura et al. 2012). The ITS (OM698813-15), tub2 (OM810165-67) andtef1α(OM810186-88) sequences were 99.38%, 99.09%, and 99.17% identical to the type N.clavispora strain MFLUCC12-0281 (accession nos. JX398979, JX399014, and JX399045) through BLAST analysis, respectively. A phylogenetic tree was generated using the concatenated sequences of ITS, tub2, and tef1α. The result showed that these three isolates were clustered with N.clavispora strains including the type MFLUCC12-0281. To perform pathogenicity tests, 20 healthy potted seedlings of A.corniculatum (2-year-old) were selected. Ten surface-sterilized leavesof 10 seedlings were wounded and inoculated by spraying conidial suspension ( 105conidia/ml). The same number wounded leaves of the other 10 plants treated with sterile water served as controls. All plants were wrapped in polyethylene bags for 24 h and incubated at 28°C in a growth chamber (at 90% relative humidity). After 10 days, all the inoculated leaves showed similar symptoms to those observed in the field, whereas control leaves were asymptomatic. N. clavispora was reisolated from the lesions in terms of morphology and molecular characterization, whereas no fungus was isolated from the control leaves. The pathogenicity test was repeated three times under the same conditions. Thus, Koch's postulates were fulfilled. This pathogen has been reported on a wide host range worldwide, such as leaf spot on strawberry in China (Zhao et al. 2016) and twig blight on blueberry in Spain (Borrero et al. 2018). This is the first report of N. clavispora causing leaf blight on A.corniculatum in China.This study provides valuable information for the identification and control of the disease.

Identification of Fusarium solani f. sp. cucurbitae Causing Zucchini Fruit Rot in Inner Mongolia, China

Fruit rot is one of the major diseases impacting the production and quality of zucchini (Cucurbita pepo). In August 2021, fruit rot symptoms were observed on the zucchini fruit ‘Jindi 1’ in the Wuyuan region in Inner Mongolia, China with an incidence ranging from 10% to 30%. Where the pepo was in contact with the soil, dark grey and spongy corky lesions 4–5 cm in diameter with a light brown halo were observed. The internal necrosis of the fruit rind was also recorded. From the affected fruits, fungal colonies belonging to the Fusarium species were exclusively isolated. Molecular analysis of the ITS, TEF-1α, and RPB2 sequences identified the isolates as Fusarium solani f. sp. cucurbitae. Inoculated on ‘Jindi 1’, the strain Fx-1a induced typical fruit rot on the pepo and wilting on seedlings, while negative-controls remained asymptomatic. The impact of this disease on seed quality and yield in zucchini seed production needs to be further studied.

Ashy Dermatosis: A Narrative Literature Review

Ashy dermatosis (AD) or erythema dyschromicum perstans (EDP) is a type of hyperpigmented macules characterized by asymptomatic, gray macular lesions with a symmetrical distribution. Some of the predisposing factors for ashy dermatosis are intestinal parasitic infections, a bad environment such as consumption of ammonium nitrate, X-ray contrast media and genetic factors such as the HLA-DR4 allele. The clinical manifestations of ashy dermatosis are hyperpigmented or gray macules with slow progression and pigmentation abnormalities in the inner epidermal layer. The differential diagnosis of ashy dermatosis is with lichen planus pigmentosus, idiopathic eruptive macular pigmentation, and Riehl's melanosis. Diagnosis of ashy dermatosis needs to be linked between clinical findings and histopathological examination, other supporting examinations and differentiating it from other differential diagnoses. Management for patients with ashy dermatosis is with topical or systemic agents. For now, a combination of laser and tacrolimus ointment is recommended.

Multimodal imaging features of acute macular retinopathy in patients with COVID-19

Objective: To investigate the multimodal imaging characteristics of acute macular retinopathy (AMR) and/or parafoveal acute middle maculopathy (PAMM) in patients with coronavirus disease 2019 (COVID-19). Methods: It was a cross-sectional study. Eight patients (15 eyes) diagnosed with AMN and/or PAMM, who presented for their initial visit at Kaifeng Eye Hospital between December 17 and December 31, 2022 and were also confirmed positive for COVID-19, were enrolled as the observation group. The patients were classified into four types based on swept-source optical coherence tomography (SS-OCT) findings. Fifteen healthy volunteers (15 eyes) without ocular or systemic diseases were recruited as the healthy control group, and one eye was randomly selected for analysis. All participants underwent detailed ophthalmic examinations, including best-corrected visual acuity (BCVA), slit-lamp biomicroscopy, fundus photography (FP), intraocular pressure measurement, fundus infrared imaging, OCT and OCT angiography (OCTA). The foveal avascular zone (FAZ) area of the macular center was measured. General information and multimodal imaging findings were collected and analyzed. The superficial capillary plexus vessel density (SCP-VD) and deep capillary plexus vessel density (DCP-VD) were measured in circular areas with diameters of 1.0 mm, >1.0 mm and ≤3.0 mm, and>3.0 mm and ≤6.0 mm centered on the foveal center, recorded as SCP-VD1.0, 3.0, 6.0 and DCP-VD1.0, 3.0, 6.0. Statistical analyses were performed using t-tests, Mann-Whitney U tests, and chi-square tests. Results: The observation group consisted of 6 males (11 eyes) and 2 females (4 eyes) with a mean age of (26.87±11.56) years. The healthy control group included 11 males (11 eyes) and 4 females (4 eyes) with a mean age of (28.75±12.30) years. There were no statistically significant differences in age and gender distribution between the two groups (all P>0.05). All patients in the observation group experienced high fever (≥39.0 ℃) and developed ocular symptoms during the febrile period or within 24 hours after fever resolution. Among all patients, there were 5 cases (7 eyes) of Type Ⅰ, 1 case (1 eye) of Type Ⅱ, 3 cases (4 eyes) of Type Ⅲ, and 2 cases (3 eyes) of Type Ⅳ. In Type Ⅲ and Ⅳ, 3 cases (4 eyes) exhibited weakly reflective cystic spaces in the outer plexiform or outer nuclear layers, and fundus photography revealed multiple gray or reddish-brown lesions in the macular region. One case (1 eye) showed retinal superficial hemorrhage. Cotton wool spots were observed in 2 cases (4 eyes). Fundus infrared imaging showed that Type Ⅰ manifested as weak reflectivity lesions in the parafoveal central zone, with the tip pointing towards the fovea. Type Ⅱ showed no apparent abnormalities in the macular region, while Type Ⅲ and Ⅳ displayed map-like weak reflective lesions spanning the foveal center. OCTA findings demonstrated that SCP-VD1.0 in the observation group was 6.93% (4.77%, 6.93%), significantly lower than the healthy control group's 10.66% (8.05%, 10.55%) (U=174.00, P=0.016). SCP-VD3.0 in the observation group was 37.14% (32.15%, 43.48%), also lower than the healthy control group's 43.06% (38.95%, 46.55%) (U=174.00, P=0.016). DCP-VD3.0 in the observation group was 48.20% (46.11%, 50.33%), lower than the healthy control group's 51.10% (50.04%, 53.02%) (U=188.00, P=0.009). DCP-VD6.0 in the observation group was 49.27% (47.26%, 51.67%), lower than the healthy control group's 52.43% (50.07%, 53.82%) (U=70.00, P=0.004). There were no significant differences in SCP-VD6.0 and DCP-VD1.0 between the two groups (both P>0.05). Conclusions: Acute macular retinopathy in patients with COVID-19 can involve all retinal layers and present as segmental hyper-reflectivity on SS-OCT. Fundus infrared imaging reveals weak reflectivity in the affected area, fundus photography shows multiple gray or reddish-brown lesions in the macular region, and OCTA demonstrates a decrease in SCP-VD and DCP-VD.

First report of Fusarium proliferatum causing leaf spot on tea plants in Jiangxi Province, China.

Tea (Camellia sinensis (L.) O. Kuntze), a perennial evergreen shrub, is one of the most important cash crops in China. In September 2021, leaf spot symptoms were observed on approximately 30% of tea plants in a 2 ha commercial field of Lushan (29°33'0" N, 115°58'48" E), Jiangxi Province, China. The symptoms initially appeared as small, gray lesions, and later became larger (10-15 mm in diameter) circular to irregular spots with light brown centers and gray borders. To isolate the pathogen, small pieces (3×3 mm) cut from the margins of lesions were sterilized with 75% ethanol for 10 s, 0.1% HgCl2 for 20 s, and then rinsed three times with sterile water. The pieces were placed onto acidified potato dextrose agar (APDA) plates, and incubated in darkness at 28℃. Pure cultures were prepared by subculturing hyphal tips. A total of 16 fungal isolates were obtained, and the colonies of 15 isolates (isolation rate 93.8%) looked identical, resembling those of the genus Fusarium. The colonies were white to pink with purple woolly mycelium. After 10 to 15 days incubation, slightly curved macroconidia with three to four septa measuring 14.0 to 34.5 × 2.0 to 3.5 µm (n = 50), and oval, unicellar microconidia measuring 4.0 to 9.0 × 1.5 to 3.5 µm (n = 50) were observed. These morphological characteristics were similar to that described for Fusarium proliferatum (Leslie and Summerell 2006). Genomic DNA of representative isolates (LSZWY, LSZWY2, LSZWY3) was extracted with the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech Co., Ltd, Shanghai). The translation elongation factor 1 alpha gene (EF-1ɑ) was amplified using primers EF-1H / EF-2T (O'Donnell, et al. 2015). PCR product was sequenced and the sequence was 709 bp (Accession No. OL614004, ON357634, ON595710). BLAST search results showed that it had 99.9% identity with the EF-1ɑ gene sequence of F. proliferatum (MH341215, MT371378). To test pathogenicity, nine leaves from 5-year-old healthy tea plants (Ca. Luyun 3) were wounded using a sterilized needle and inoculated with a 20µl conidial suspension (2 × 107 conidia·mL-1) on one side of the plants and the other side with sterilized distilled water as a control. All leaves were incubated in a growth chamber at 28℃ and 80% relative humidity with a 12 h light/dark photoperiod. Seven days later, all inoculated treatments showed symptoms identical to those observed in the field, while the control remained asymptomatic. The experiment was repeated three times with similar results. Koch's postulates were fulfilled by successful re-isolation and morphological and molecular identification of F. proliferatum from the inoculated leaves. This pathogen can cause diseases of many crops, e.g. tobacco, Polygonatum cyrtonema and others (Li, et al 2017; Zhou, et al. 2021). However, this is the first report of F. proliferatum causing leaf spot on tea plants in China. This new disease poses a threat to the yield and quality of tea and methods need to be developed for its control and to prevent further spread.