Translation Elongation Factor 1-alpha Research Articles

First Report of Agroathelia rolfsii Causing Southern Blight on Alpinia hainanensis in China.

Katsumada galangal seed (Alpinia hainanensis K. Schum) is an important member of the Zingiberaceae family, with both medicinal value and culinary applications (Park et al. 2020). In May 2023, a stem disease of Al. hainanensis was detected in a 100-acre plantation located in Danzhou City, Hainan Province in China, with an average incidence of 15% to 20%. The disease primarily targeted the basal stems, with the pathogen spreading by mycelia in the soil around the rhizome, and reaching neighboring plants. The infection caused the stem base to undergo necrosis, turning the tissue brown to black, and leading to a loosely attached epidermis. As the disease advanced, the necrosis progressed upward along the stem, resulting in curling of the leaves. A large number of white mycelia, as well as white immature sclerotia and brown mature sclerotia, were also observed on the surfaces of the leaves, stems and soil. The diameter of the mature sclerotia ranged from 0.7 to 1.6 mm (aver. ± SD = 1.2 ± 0.3 mm, n = 50). To identify the pathogen, eleven tissue samples were collected from symptomatic stems. The sclerotia on the surface of the tissue were sterilized with 75% (V/V) ethanol for 30 s, followed by three rinses with sterile water, and finally incubated on potato dextrose agar (PDA) at 28°C in darkness. Eleven isolates were obtained and subcultured at 28°C in darkness. After 48 hours, the colonies of these isolates had an average daily radial growth of 39.5 ± 0.4 mm (n = 11) and were white, featuring entire margins and abundant aerial mycelia. By the seventh day, spherical or oval white sclerotia began to form on the surface of their colonies. These sclerotia eventually turned brown, with a diameter ranging from 0.6 to 1.5 mm (aver. ± SD = 1.3 ± 0.3 mm, n = 50). The morphological characteristics of these isolates were similar to those of Agroathelia rolfsii (Sacc.) Redhead & Mullineux (basionym: Sclerotium rolfsii Sacc.; Amylocorticiaceae) (Redhead and Mullineux 2023). DNA of a representative isolate DZAS-01 was extracted for amplification of its internal transcribed spacer (ITS) rDNA region, the translation elongation factor 1-alpha (TEF1) gene and the large subunit (LSU) region using the primers ITS1/ITS4, EF1-983F/EF1-2218R and LROR/LR5, respectively (Moncalvo et al. 2000, Rehner and Buckley 2005, White et al. 1990). The three sequences have been deposited in GenBank (accession nos. PP659527 for ITS, 607 bp; PP976301 for TEF1, 1,040 bp; PP968104 for LSU, 876 bp). The BLASTn results showed that the similarity of the three sequences with the known sequences PP908473 (ITS), OL416131 (TEF1) and KY446370 (LSU) of Ag. rolfsii was 100%, 99.81% and 100%, respectively. Pathogenicity tests were conducted on ten potted, 6-week-old healthy Al. hainanensis plants. The stem bases of ten plants were wounded with sterile needles and divided into two groups with five plants in each group. Each plant in the first group was inoculated by placing three mature sclerotia of DZAS-01 near the wound (Urbina et al. 2024). Plants in the other group served as controls and were not inoculated. Ten plants were covered with plastic bags, placed at 25°C to 28°C with a 12-h/12-h light/dark cycle, and sprayed with sterile water daily. After seven days, all inoculated plants exhibited symptoms that were consistent with those in the field, while non-inoculated control plants remained symptom-free. This pathogenicity test was repeated three times with identical results. A fungus was re-isolated from symptomatic tissues of inoculated plants in each repeat trial and had identical morphological characteristics and ITS, TEF1 and LSU sequences to those of isolate DZAS-01. Ag. rolfsii can infect more than 500 species of cultivated and wild plants in tropical and subtropical regions (Daunde et al. 2020). To the best of our knowledge, this is the first report of Ag. rolfsii causing southern blight on Al. hainanensis in China. The results obtained from this study are going to provide guidance for preventing and treating this disease.

First report of Ilyonectria liriodendri causing black foot on walnut in Chile.

Walnut (Juglans regia L.) is the primary nut tree cultivated in Chile, covering 44.626 ha. In autumn 2021, walnut decline and tree mortality were observed in young and old orchards (two and 18-years-old, respectively) in the O´Higgins Region in the Chilean Central Valley. In the surveyed walnut orchards cv. Chandler (n = 2), the incidence of symptomatic plants ranged from 35 to more than 90 %. Affected trees showed poor growth, twig and branch death, early defoliation, and shoot dieback. In addition, collar base necrosis and root rot were observed in symptomatic trees, resembling the symptoms of black foot disease, with blackening of complete roots and even the entire root system. Symptomatic root tissues from 5 independent trees were cut into pieces of about 25 mm2. The samples were sterilized for three minutes in a sodium hypochlorite solution (1%), rinsed in sterile distilled water, and plated onto Potato Dextrose Agar (PDA) medium supplemented with lactic acid (10%) and 0.5 g/L of streptomycin sulfate. Mycelium grew from the samples after seven days of incubation at 24°C. The characteristics of the culture and morphology of conidiophores and conidia indicated a Cylindrocarpon-like fungus. Then, 5 single-conidial isolates were obtained and subsequently grown on a PDA medium. Genomic DNA was extracted from cultures of three representative isolates (CNCO5; CNCO6; CNSF2) and used for amplification and sequencing of the Internal Transcribed Spacer of rDNA (ITS), β-tubulin (TUB2), Histone3 (HIS3), and Translation Elongation Factor 1-alpha (TEF1-α) partial gene regions using the primers ITS1/ITS4 (White et al. 1990), T1/Bt2b (O'Donnell and Cigelnik 1997; Glass and Donaldson 1995), CYLH3F/CYLH3R (Crous et al. 2004), and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively. Isolates clustered consistently with Ilyonectria liriodendri after a multilocus molecular phylogenetic analysis. Sequences were deposited in at NCBI Genbank data base (ITS: OR871536-OR871537-OR871538, TUB2: PP780828-PP780829-PP780830, HIS3: PP780822-PP780823-PP780824, and TEF1-α: PP780825-PP780826-PP780827). A pathogenicity test was conducted by inoculating three walnut Vlach rootstocks (six months old) with one representative I. liriodendri isolate (CNCO6). Plant roots were cut 5 cm from the root tip and immersed in a 105/mL conidial suspension for 30 min; control plants were immersed in sterile distilled water. Then, the plants were transplanted to a sterile substrate. After six weeks of growing in a greenhouse, necrosis characterized by brown-black coloration of the roots was observed developing upwards from the wounded roots. Necrotic lesions were found even in secondary roots of infected plants; however, control plants remained asymptomatic. The fungus was re-isolated, completing Koch's postulates. Black foot disease caused by Ilyonectria spp. has been reported worldwide on grapevines (Ye et al., 2021) and causing root rot of other fruit trees such as olive (Úrbez-Torres et al., 2012) and kiwifruit (Erper et al., 2011). Ilyonectria liriodendri was recently identified as the cause of walnuts root rot in California (Lawrence et al. 2019). To our knowledge, this is the first detection of I. liriodendri causing black foot of walnut in Chile and worldwide. The symptoms of this disease closely resemble those of Phytophthora spp. in walnuts. This similarity could lead to misdiagnosis, highlighting the necessity for specific diagnostic tools to accurately identify the pathogen.

First Report of Neoscytalidium dimidiatum Causing Fruit Rot on Fig in China.

Fig (Ficus carica L.) holds economic significance in Atushi, Xinjiang, but as fig cultivation expands, disease prevalence has risen. In July 2024, approximately 22% of harvested fig (cv. Xinjiang Zaohuang) from 20 commercial orchards (covering 40 hectares) in Atushi (39°39'37.65" N, 76°14'3.62" E) showed varying degrees of fruit rot symptoms. The initial symptoms were characterized by the appearance of small, brown lesions on the fruit surface. These lesions rapidly progressed into water-soaked spots, which expanded quickly. As the disease advanced, the affected areas became covered with dense, white, fluffy mycelia, accompanied by prominent black sporulation. In later stages, the infected tissues softened further, ultimately resulting in the complete decay of the fruit. Twenty diseased fig were collected from the sampling site. Tissue samples (5×5×5 mm) were cut at the diseased-healthy junction, surface-sterilized in 0.5% NaClO for 1 minute, rinsed twice in sterile distilled water, air-dried, and transferred aseptically onto potato dextrose agar (PDA), and incubated at 25°C for 5 days with a 12-hour photoperiod. Fifteen isolates were obtained from the infected tissues, with two representative isolates (WH 12 and WH 23) selected for further study due to morphological similarity. The fungal colonies initially appeared as white mycelium, later turning olive green to grayish-black. Colony growth was rapid (32 mm/day). Arthrospores were colorless to light brown, short columnar, aseptate, with a truncated base, 0 to 1 septate, averaging 11.9±2.3×3.6±0.8 μm (n = 50), and sometimes formed arthric chains. Chlamydospores were dark brown, round or oval, 0 to 1 septate, averaging 7.26±1.7×5.05±1.0 μm (n = 50). Genomic DNA was extracted from the two isolates. The internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF1-α), and beta-tubulin (TUB2) genes were amplified using primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone & Kohn. 1999), and BT2a/BT2b (Glass & Donaldson. 1995), respectively, and sequences were deposited in GenBank (ITS: PQ555020, PQ555021; TUB2: PQ557519, PQ557521; TEF1-α: PQ557520, PQ557522). BLAST analysis revealed 99-100% similarity to Neoscytalidium dimidiatum Arp2-D (ITS: MK813852; TUB2: MK816354; TEF1-α: MK816355). Phylogenetic analysis using IQ-Tree and MrBayes3.2.7 based on concatenated ITS-TEF1-TUB sequences showed WH 12 and WH 23 clustering with N. dimidiatum Arp2-D (99% bootstrap). Morphological and molecular data identified the isolates as N. dimidiatum (Penz.) Crous & Slippers (Crous et al. 2006). Pathogenicity tests were conducted on 20 healthy fig (cv. Xinjiang Zaohuang) by inoculating each fruit with 10 µl of a WH 12 conidial suspension (1 × 10⁶ conidia/ml) using sterile needles. The Control were treated with 10 µl of sterile distilled water. All fruits were placed in sterile plastic containers and incubated at 25 ± 1°C, 90% relative humidity, and a 12-hour light cycle. This experiment was performed twice. On the 1st day post-inoculation, brown lesions began to develop on the fruit. By the 4th day post-inoculation, the entire fruit was completely decayed and covered with white mycelia and black spores, while the control fruit showed no symptoms. The fungus was successfully reisolated from the inoculated fruits and identified as N. dimidiatum following the methods described above, fulfilling Koch's postulates. N. dimidiatum has been reported to have a wide range of hosts in China, such as Jacaranda mimosifolia, Hylocereus megalanthus, Hylocereus undatus, and Styphnolobium japonicum (Li et al. 2024; Zeng et al. 2024; Lan et al. 2022; Luo et al. 2024). To our knowledge, this study is the first report of N. dimidiatum as the causal agent of fruit rot in fig in China. Our findings have expanded the host range of N. dimidiatum in China and provide a theoretical basis for the diagnosis and treatment of the disease.

Isolation and Identification of Apiospora intestini from Hedera nepalensis Leaf Blight and Determination of Antagonism of Phomopsis sp.

In order to identify the pathogen responsible for Hedera nepalensis leaf blight and investigate effective biocontrol strategies, samples were collected from 10 significantly infected areas at Southwest Forestry University; four to six infected leaves were gathered from each area, followed by the isolation and purification of strains from the infected plant leaves using tissue isolation and hyphae-purification techniques. We conducted an examination of the biological characteristics and compared the inhibitory effects of different concentrations of Phomopsis sp. (50%, 25%, 16.7%, 12.5%, and 10%) with 20 µg/mL of synthetic fungicides (Mancozeb, Carbendazim, Polyoxin, and Hymexazol) on the pathogen, while also assessing the control efficacy of Phomopsis sp. against the pathogen in the greenhouse. The internal transcribed spacer (ITS) region, β-tubulin (TUB), and translation elongation factor 1-alpha (TEF) analysis revealed that the highly virulent strain causing H. nepalensis leaf blight was Apiospora intestini. Additionally, it was found that 25% Phomopsis sp. significantly inhibited Apiospora intestini when compared to synthetic fungicides, and Phomopsis sp. supernatant possesses both protective and curative effects against the plant diseases caused by Apiospora intestini. The results of this study serve as a reference for the prevention and treatment of H. nepalensis leaf blight.

Aggressiveness and phylogenetic relationship of Fusarium oxysporum associated with crown and root rot in pyrethrum plants.

In Australia, pyrethrum (Tanacetum cinerariifolium) cultivation provides a significant portion of the global supply of natural insecticidal pyrethrins. However, crown and root rots, along with stunted plant growth and plant loss during winter, are significant issues affecting certain sites. Several isolates of the Fusarium oxysporum species complex (FOSC) have been identified as causal agents of crown and root rot in pyrethrum, highlighting these as key pathogens contributing to this decline. However, the genetic and pathogenic diversity of the FOSC impacting pyrethrum is unclear. This study isolated F. oxysporum consistently from symptomatic and asymptomatic field-grown pyrethrum plant tissues, identified through morphological and multigene phylogenetic analyses. Phylogenetic analyses of partial gene sequences of calmodulin (cmdA), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1-α) and β-tubulin (tub2) resolved the placement of these isolates within the context of different published FOSC taxonomies and revealed notable genetic diversity among the isolates. Glasshouse experiments effectively reproduced the crown and root rot symptoms observed in field conditions, demonstrating a similar level of aggressiveness among F. oxysporum isolates from pyrethrum plants. The results indicate the importance of soil-borne disease management to reduce yield decline in pyrethrum fields and will help with the selection of aggressive isolates for resistance screening of pyrethrum varieties.

Morpho-phylogenetic analyses of two novel edible mushrooms from China and a mini review of Lyophyllum (Agaricales, Lyophyllaceae) cultivation and bioactivities

Lyophyllum plays an important role in the natural ecosystem and has significant economic value. Some species of this genus have been cultivated in Asia, America, and Europe. This study describes four edible species of Lyophyllum, two of which were newly discovered. Lyophyllum edulis has a dark grayish orange pileus, a grayish orange upper part of the stipe, and globose, subglobose to broadly ellipsoid basidiospores, while L. sinense has a dark gray-orange when injured pileus, dark grayish orange points and lines on the stipe surface, and quadrangular to broadly fusiform basidiospores. Molecular phylogenetic analyses using the internal transcribed spacer ITS1-5.8S-ITS2 ribosomal RNA (ITS), the large subunit ribosomal RNA (LSU), the second-largest subunit of RNA polymerase II (rpb2), and translation elongation factor 1-alpha (tef1-α) indicated that L. edulis is related to L. shimeji, L. heimogu, and L. decastes, and L. sinense has an affinity to L. bulborhizum and L. nigrum. We also summarize the cultivation techniques of the two edible species, L. shimeji and L. decastes.

Morpho-phylogenetic evidence reveals novel hyphomycetous fungi on medicinal plants in Southwestern China

ABSTRACT During a survey of saprobic fungi on medicinal plants in Southwestern China, thirty-nine hyphomycetous collections belonging to Dictyosporiaceae, Melanommataceae, and Stachybotryaceae were identified, representing nineteen distinct species. These taxa were characterised and identified based on morphological and culture characteristics, coupled with phylogenetic analyses of combined sequences of calmodulin (cmdA), the internal transcribed spacer region of ribosomal DNA (ITS), nuclear large subunit ribosomal DNA (LSU), RNA polymerase second-largest subunit (rpb2), the translation elongation factor 1-alpha (tef1-α), and β-tubulin (tub2). Twelve novel species are described, including Camposporium alangii, C. polygoni, Dendryphiella verrucosispora, Jalapriya cheirospora, Memnoniella cnidiicola, M. guttulatispora, M. reniformis, M. reynoutriae, M. verrucosispora, Sirastachys aspidistrae, Sir. ellipsoidispora, and Striatibotrys biguttulatispora. Additionally, three new host records, M. alishanensis, Sir. castanedae, and Stachybotrys chartarum, and four new hosts and geographical records of M. ellipsoidea, M. pseudonilagirica, Str. rhabdospora, and Virgatospora echinofibrosa are reported. Memnoniella nilagirica is revised and synonymised under M. pseudonilagirica. Key morphological characteristics, hosts specificity, and distributional data of Memnoniella, Sirastachys, and Striatibotrys were summarised. This study provides comprehensive illustrations, descriptions, and notes for each new taxon and record, marking the first report of these species from medicinal plants.

Morphological and phylogenetic analyses reveal two new species of the Fusariumfujikuroi (Hypocreales, Nectriaceae) species complex in China.

The Fusariumfujikuroi species complex (FFSC) encompasses a diverse array of more than 80 phylogenetic species with both phytopathological and clinical importance. A stable taxonomy is crucial for species in the FFSC due to their economical relevance. Fungal strains used in this study were obtained from Castaneamollissima and Rubuslambertianus, collected from Beijing and Shaanxi Province. We employ morphological and phylogenetic analyses based on partial gene fragments of the translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), calmodulin (CaM), RNA polymerase largest subunit (rpb1), and RNA polymerase II second largest subunit (rpb2), as well as the pairwise homoplasy index tests. Studies have shown that these phylospecies are clustered in the Asian clade of the FFSC. The present study delineates two novel Fusarium species within the FFSC, named F.castaneophilum and F.rubicola, complemented by illustrations and descriptions.

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

During a field study in the Baili Azalea Forest Area in Guizhou Province, China (27°12'N, 105°48'E) between May and July 2023, symptoms of leaf spot were observed on Rhododendron latoucheae Franch. The incidence of leaf spot on R. latoucheae leaves was about 12% in a field of 1 hm2, significantly reducing their ornamental and economic value. The affected leaves bore irregular, grey-white lesions with distinct dark brown borders, accompanied by black conidiomata. To isolate the pathogen, 15 symptomatic leaves were collected from 10 plants. Conidiomata present on the lesions were transferred onto water agar and incubated at 25℃ for 24 hours. Germinated spores were then transferred onto potato dextrose agar (PDA) to generate single spore cultures for morphological analysis. This process yielded three single-spore isolates: GZULJ 3-6.1, GZULJ 3-6.2, and GZULJ 3-6.3, all displaying identical morphological characteristics. Representative isolate GZULJ 3-6.1 was used for further study. The colonies, cultivated on PDA at 25°C in the dark, appeared pale gray with white aerial mycelium. Alpha conidia were single celled, translucent and fusiform, measuring 6.2 to 8.1 × 2.6 to 3.0 μm (n=50). Beta conidia were filamentous and hook-shaped, measuring 17.0 to 23.0 × 0.9 to 1.5 μm (n=50). The morphological features were consistent with the established description of Diaporthe hongkongensis (Zhang et al. 2021). For molecular identification, total genomic DNA was extracted. The complete internal transcribed spacer region (ITS), partial sequence of beta-tubulin (TUB), translation elongation factor 1-alpha (TEF1), and calmodulin (CAL) gene were amplified and sequenced using primers ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass and Donaldson. 1995), EF1-728F/EF1-986R and CAL-228F/CAL-737R (Carbone and Kohn. 1999), respectively. The ITS, TUB, TEF1, and CAL sequences (GenBank accession nos. OR807761, OR825453, OR825450, and OR825447, respectively) were obtained, and BLASTn analysis against sequences in GenBank showed 97.37 to 99.30% identity with Diaporthe hongkongensis CBS 115448. Phylogenetic analysis reaffirmed the isolate's placement within a well-supported cluster alongside D. hongkongensis. The pathogenicity of GZULJ 3-6.1 was evaluated using three 2-year-old R. latoucheae plants inoculated with spore suspension (106 spores per ml). The inoculation targeted three leaves per plant, while an additional three plants, serving as controls, were sprayed with sterile water. The plants were maintained in the growth chamber at 25°C, with a 12-hour photoperiod and 75% relative humidity. The pathogenicity test was repeated three times. After 14 days, the inoculated leaves developed brown lesions similar to those observed in the field, whereas the control leaves remained symptom-free. The fungus was successfully reisolated from the infected leaves and identified through morphological characterization and molecular analyses, confirming its identity. D. eres, a closely related species to D. hongkongensis, has been reported to induce similar disease symptoms on R. latoucheae (Wu et al. 2024). However, to our knowledge, this is the first report of D. hongkongensis causing leaf spot disease on R. latoucheae in China. This finding will facilitate the development of effective management strategies to control the complex leaf spot disease affecting R. latoucheae and provide critical insights into its broader implications for the health of this species.

First Report of Botryosphaeria dothidea Causing Fruit Rot on Hawthorn (Crataegus pinnatifida) in China.

Hawthorn (Crataegus pinnatifida) is an important economic fruit and Chinese medicinal plant, which is widely distributed in the northern China. In early July 2024, a fruit rot disease was observed on the young fruits of hawthorn in a park of Shouguang, Shandong Province, China (36°53'42.16″N, 118°47'22.4″E). Forty plants were surveyed (from approximately 120 trees), which had 15 to 30% of fruits infected. Diseased fruits displayed brown, sunken, approximately circular lesions. Small sections (3-4 mm) cut from the edges between the diseased and healthy tissue from 20 diseased fruits were surface sterilized by using 75% ethanol for 30 s, and rinsed thrice with sterile distilled water. The diseased tissues were then dried and plated onto potato dextrose agar (PDA) and incubated at 25°C in the dark. Cultures were purified by hyphal tipping. In total, 12 colonies were isolated from the necrotic tissues, and all of them showed similar morphological characteristics. The colonies were initially white, gradually turning gray-green to dark gray after 5 days. Conidia were hyaline, aseptate, ellipsoidal to obovoid, 20.1-29.3 × 5.0-8.3 μm, with an average of 24.2 × 6.6 μm (n = 30). These morphological traits suggested that the pathogen shares similarities with the Botryosphaeria dothidea (Zhang, et al., 2021). For accurate identification, three representative isolates (SGSZ-01 to SGSZ-03) were selected for molecular identification by amplifying and sequencing the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF1-α), and β-tubulin (TUB2) with the primers ITS1/ITS4, EF1-728F/986R, and Bt2a/Bt2b (Carbone and Kohn 1999; Jia et al. 2019), respectively. GenBank accession numbers were PQ197601, PQ586368, and PQ586369 for ITS, PQ201924, PQ588676, and PQ588677 for TEF1-α, and PQ201925, PQ588678, and PQ588679 for TUB2, and gene sequencing showed 99.2 to 100% identity with the ex-type strain of B. dothidea (CMW 8000). Results from the maximum likelihood phylogenetic analysis, based on the three concatenated gene sequences, placed our isolates in a clade with B. dothidea. The pathogenicity of SGSZ-01 was conducted using a wound inoculation method on 11-week-old fruits of 10-year-old hawthorn trees both on living plants and detached fruits. All fruits were wounded with a sterilized needle and inoculated by placing 5 mm2 agar plugs of 5-day-old cultures grown on PDA. Control fruits were treated with agar plugs. All the attached fruits were enclosed in a plastic bag with a wet cotton ball for 5 days. The same number of detached inoculated fruits were placed in Petri dishes on a piece of wet filter paper and incubated at 25°C. The pathogenicity test was conducted three times independently. Necrotic lesions (4.5 ± 0.3 mm in diameter) resembling natural infections were observed on fruits 7 days after inoculation, and no symptoms appeared on the control fruits. The pathogen was reisolated from the lesions and was identified by the methods described above. No pathogens were isolated from control fruits. Therefore, the pathogen of hawthorn fruit rot in China is B. dothidea. B. dothidea has previously been reported causing fruit rot on mango, plum, pomegranate fruit, and apple in China (Feng et al. 2023; Gu et al. 2020; Tang et al. 2012; Yuan et al. 2024). To our knowledge, this is the first report of B. dothidea causing fruit rot disease on C. pinnatifida. The information on identification of this fungus may be helpful to the control and prevention of the disease.

Pseudocercospora fengxinensis, a new species causing sooty spot of kiwifruit in China.

Pseudocercospora is a large cosmopolitan genus of plant pathogenic fungi that are commonly associated with leaf and fruit spots as well as blights on a wide range of plant hosts. P. actinidiae is a member of this genus, causing sooty spot on kiwifruit worldwide. With the expansion of kiwifruit cultivation, the incidence of sooty spot has become severe in Fengxin County, Jiangxi Province, China. Based on a survey conducted in 2022 across nine kiwifruit cultivars in six local orchards, the results showed that the disease incidence was higher in 'Donghong', 'Jinyan', 'Jinkui', 'Fenghuang No. 1' and 'Jinguo'. In contrast, it was lower in 'Gold3' and 'Hongshi No. 2'. No lesions were observed on 'Kuilv' and 'Miliang No. 1'. 31 fungal strains were isolated from infected leaves using single-spore isolation. Phylogenetic analysis of combined sequence data from the internal transcribed spacer region of ribosomal DNA (ITS), partial actin (actA), translation elongation factor 1-alpha (tef1), and the second largest subunit of RNA polymerase II (rpb2) sequence data, using maximum-likelihood (ML) and Bayesian inference (BI), revealed their taxonomic placement within Pseudocercospora. Both morphological and molecular phylogenetic analysis supported Pseudocercospora fengxinensis as a new taxon within Pseudocercospora. Pathogenicity tests confirmed P. fengxinensis as the causal agent of sooty spot on kiwifruit. This study is the first report of P. fengxinensis causing sooty spot of kiwifruit worldwide, enriching our understanding of Pseudocercospora diversity in China and providing a foundation for improved disease management strategies.

First Report of Exserohilum rostratum Causing Leaf Blight on Calathea orbifolia in China.

Peacock plant (Calathea orbifolia (Linden) H.A.Kenn.) is cultivated as a valuable houseplant in China. In September 2024, leaf blight was found on this plant in a field located in Shenzhen (22°35'55"N, 113°59'21"E), Guangdong Province, China. The area investigated was ~900 m2. The disease incidence was ~70% of 100 plants. Initial symptoms of brown spots appeared on leaves, gradually becoming enlarged, elongated, 0.5 to 10 × 0.2 to 4.5 cm, irregular, dark brown, with a yellow halo, and leaves blighted and desiccated. Ten symptomatic leaves were collected for pathogen isolation. Leaf pieces (5×5 mm2) of lesion margins were surface disinfected with 1% NaClO for 1 min, 70% ethanol for 30 s, rinsed in sterile distilled water, placed on PDA, and incubated at 25°C. After 3 days, fungal colonies were purified by a single hyphal tip method on PDA. Three isolates (MBSZU 24-001 to MBSZU 24-003) with similar morphology were obtained. Colonies on PDA were 48 to 54 mm in diameter after 1 week at 25°C, then gray, flat with entire edges, aerial mycelia dense, and dark in the center and gray at the margin on the reverse side after 2 weeks. Conidiophores were olivaceous-brown, cylindrical, septate, unbranched, up to 230 × 3.4 to 5 µm. Conidiogenous cells were integrated, straight to curved, proliferating sympodial, 25 to 60 × 4.5 to 7 µm. Conidia were straight to slightly curved, ellipsoid, pale olivaceous brown to dark olivaceous brown, 2-6-distoseptate, with a distinct basal hilum, 26.9 to 70 × 11.4 to 21.6 µm (n = 50). Morphological characteristics of all isolates resembled the Exserohilum (Hernández-Restrepo et al. 2018). The internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1-alpha (tef1-α) genes were amplified using primer pairs ITS5/ITS4 (White et al. 1990), GPD1/GPD2 (Berbee et al. 1999), and EF1-983F/EF1-2218R (Rehner and Buckley 2005), respectively. Sequences were deposited in GenBank (ITS: PQ498876 to PQ498878; GAPDH: PQ509760 to PQ509762; tef1-α: PQ509763 to PQ509765). In the BLAST analysis, the ITS, GAPDH, and tef1-α sequences showed 99.50% (599 bp out of 602 bp), 98.89% (536 bp out of 542 bp), and 99.78% (926 bp out of 928 bp), respectively, to E. rostratum (CBS 325.87). Maximum likelihood and Bayesian inference phylogenetic analyses of the concatenated three genes identified all isolates as E. rostratum. To test pathogenicity, both wounded and unwounded healthy leaves were used in this experiment. All leaves were wiped with 0.1% NaClO and then rinsed three times with sterile water. Conidia suspensions (15 µl of 1 × 106 conidia/ml) of each isolate grown on PDA at 25°C for 3 weeks were placed on all samples using the attached leaf assay. Control leaves were mock-inoculated with sterile distilled water. Ten replications were conducted for each treatment and repeated twice. Plants were incubated at 25°C with 80 to 85% relative humidity. After 10 days, all inoculated leaves had brown lesions with yellow halos, while control leaves remained asymptomatic. E. rostratum was consistently reisolated from the lesions on PDA to complete Koch's postulates. Prior to this study, E. rostratum was identified as a causal agent of leaf blight on Calathea picturata in China (Taiwan, Chern et al. 2011). This is the first report of leaf blight caused by E. rostratum on C. orbifolia in China and worldwide. These findings will provide insights for epidemiological investigations and future approaches to managing this disease.

Taxonomic novelty in Pleomonodictydaceae and new reports for Ampelomycesquisqualis (Phaeosphaeriaceae), Melomastiamaolanensis and M.oleae (Pleurotremataceae).

This study introduces a novel genus Robiniigena, with its type R.hyalinospora. The specimen was collected on dead aerial branches of Robiniapseudoacacia in Italy. Based on the examination of morphology and the results of phylogenetic analyses involving nuclear 18S rDNA (SSU), nuclear 28S rDNA (LSU), nuclear rDNA ITS1-5.8S-ITS2 (ITS), translation elongation factor 1-alpha (tef1-α) and RNA polymerase II second largest subunit (rpb2) sequences, Robiniigena is referred to the family Pleomonodictydaceae (Pleosporales). It is characterized by immersed to erumpent, ostiolate ascomata, filiform, septate and cellular pseudoparaphyses, bitunicate, clavate to cylindric-clavate asci and fusiform, hyaline ascospores surrounded by a mucilaginous sheath. This research also establishes the taxonomic placement of the previously unclassified Inflatispora (Pleosporales genus incertae sedis) within the Pleomonodictydaceae. The sexual morph of Ampelomycesquisqualis (Phaeosphaeriaceae) is described for the first time and it is characterized by immersed, perithecial ascomata, a peridium comprising two layers, branched, septate and filiform pseudoparaphyses, short-pedicellate, bitunicate asci with an ocular chamber and sub-hyaline, fusiform, septate ascospores. This species, previously known only in its asexual morph, has been found as a saprobe on Sonchus sp. in Italy. Our identification of the sexual morph was based on LSU rDNA and ITS rDNA sequence data. Melomastiamaolanensis (Pleurotremataceae) is reported for the first time in Thailand, collected from Chromolaenaodorata, while M.oleae is documented as a new record from Durantaerecta in Thailand.

Endophytic Diaporthe species from Brazil.

Diaporthe species can inhabit various hosts with different lifestyles and live as endophytes, pathogens, and saprobes. Our study analysed 180 endophytic Diaporthe isolates from Miconia sp. in the Atlantic Forest, Brosimum gaudichaudii in the Brazilian savanna (Cerrado), and Anacardium occidentale in the Caatinga forest and Cerrado in Brazil. Based on multi-locus phylogenetic analyses [β-tubulin (tub2), internal transcribed spacer regions and intervening 5.8S rRNA (ITS), translation elongation factor 1-alpha (tef1), calmodulin (cmdA), and histone (his3)] and morphological features, we are introducing seven new species (D. azevedoi, D. catimbauensis, D. coracoralinae, D. luizorum, D. pedratalhadensis, D. samambaiaensis, and D. vargemgrandensis) and reporting seven known species (D. fructicola, D. inconspicua, D. infertilis, D. paranensis, D. raonikayaporum, D. schini, and D. ueckeri). We also included a morphological description of D. infertilis and synonymised D. lutescens, D. pseudoinconspicua, and D. samaneae under D. inconspicua; D. neoraonikayoporum under D. raonikayaporum; and D. passifloricola, D. rosae, and D. vochysiae under D. ueckeri, based on limited nucleotide differences among DNA sequence data and overlapping morphological features. Our results highlight the importance of including endophytic isolates in the phylogeny of Diaporthe, and show how these data expand the geographic distribution and host relationships of known species. Citation: Ferro LO, Bezerra JDP, da Silva TM, de Oliveira CS, Nascimento SS, Paiva LM, Fan X, Crous PW, Souza-Motta CM (2024). Endophytic Diaporthe species from Brazil. Fungal Systematics and Evolution 14: 251-269. doi: 10.3114/fuse.2024.14.16.

Leaf spot of Acorus calamus var. angustatus Caused by Alternaria alternata in Anhui Province, China.

Acorus calamus var. angustatus Besser, a perennial herb of the Araceae family, was first reported in the ShenNong'sHerbalClassic and is widely distributed in southern China (Li 1979). It is important in traditional Chinese medicine for treating heart, stomach, and brain ailments (Lam et al. 2016). In March 2024, leaf spots were found on its leaves in a traditional Chinese medicine planting base in Yuexi County (30°91'56″ N, 116°19'24″ E), Anhui Province, with an incidence of about 35%. The symptoms began as small, light brown lesions that expanded, resulting in necrotic lesions ranging from 1 to 8 mm in diameter with brown halos. 3 × 3 mm sections, including both symptomatic and asymptomatic tissues, were cut from six infected plants. They were disinfected in 75% ethanol for 30 s, washed three times with sterile distilled water, transferred to Petri dishes containing potato dextrose agar (PDA) and incubated at 25 °C in the dark. Purified fungal isolates were obtained by the single-spore isolation method. The colonies on PDA initially appeared white, gradually became olive-green with 1 to 3 mm white margins and abundant aerial hyphae, while the reverse was greyish green to black. The conidia were light brown, ellipsoidal, obclavate, and 10.0 to 49.5 µm × 4.5 to 16.4 µm (mean 23.9 × 10.2 µm, n=50) in size, with 0 to 6 transverse septa and 0 to 6 longitudinal or oblique septa (n=50). Conidiophores were thick, dark brown, single-celled, with multiple conidial scars, measuring 12.8 to 146.8 × 2.9 to 6.1 (mean 50.6 × 4.2) µm (n=50). Based on above observations, the pathogen were identified as Alternaria spp. (Simmons 2007). Three representative isolates, SCP-1, SCP-2, and SCP-3 were selected for molecular identification. The Internal transcribed spacer (ITS), Alternaria major allergen (Alt a 1), and translation elongation factor 1-alpha (TEF1) genes were amplified with the primers ITS1/ITS4 (White et al. 1990), Alt-for/Alt-rev (Woudenberg et al. 2015) and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively. The sequences of the three isolates were consistent, and the sequences of isolate SCP-1 were submitted to NCBI GenBank (ITS, PP723104; Alt a1, PP708704; TEF1, PP708703). The ITS region of isolate SCP-1 was 100% similar to A. alternata TCS3002 (MN394880, Wang et al. 2023), the Alt a 1 gene was 100% similar to A. alternata CBS 620.83 (KP123868), and the TEF1 gene was 100% similar to A. alternata CBS 916.96 ex-type (KC584634). A phylogenetic tree based on sequences of ITS, Alt a 1 and TEF1 genes was constructed using the neighbor-joining method in MEGA 7 software, confirming the fungus as A. alternata. Three healthy plants of A. calamus var. angustatus were spayed with conidial suspension (1 × 107 conidia/ml) of isolate SCP-1. Three additional plants sprayed only with sterile distilled water were controls. All plants were covered with plastic bags to maintain a relative humidity of 90% for 48 h and incubated at 25 °C under a 12-h light photo-period. Twelve days post-inoculation, brown necrotic lesions developed on the inoculated leaves, enlarged, and the symptoms were similar to the original ones. The control plants remained healthy. The fungus was re-isolated from the infected plants and confirmed by morphological traits and molecular methods, fulfilling Koch's postulates. To our knowledge, there are no other reports of this fungus on A. calamus var. angustatus in Anhui, China. This report will help identify the disease based on field symptoms and provide a basis for disease management strategies of A. calamus var. angustatus.

Occurrence of Diplodia seriata causing leaf blight on Aucuba japonica in China.

Aucuba japonica is widely planted in China for landscaping purposes, often used for decoration in gardens and parks. In October 2023, a leaf blight on A. japonica was observed in Meicheng Park of Nanyang City (32°59'21″ N, 112°32'54″ E), Henan province. Subsequently, surveys were conducted in different sections of the park, a plant diseases incidence rate of 52% (n = 100). Initial symptoms included black spots and enlarged to leaf blight lesions on leaves. These lesions subsequently spread throughout the plant, causing defoliation, and aesthetic damage. Twenty diseased leaves from 15 plants were collected, and sections between the symptomatic and healthy tissues were cut into 3 × 3 mm2 pieces to isolate the pathogen. The disinfestation process involved surface sterilization with 75% ethanol solution for 30 s, followed by 1% NaClO solution for 1 min., then rinsing with sterile water, and placing on potato dextrose agar (PDA) plates, incubating at 28°C for 5 days under a photoperiodic condition of 12-hour light and 12-hour dark. After isolation, purified fungal isolates (n = 28) were obtained from 33 samples plated. These fungal isolates showed similar morphological phenotypes. The three isolates (DS2, DS8, and DS28) from different park areas were selected for subsequent identification. The colonies were gray to dark brown with abundant aerial mycelium on the surface. For sporulation, isolates were cultivated on 2% water agar bearing autoclaved poplar twigs at 25°C for 21 days (Phillips et al. 2007). Immature conidia were oblong to ovoid with rounded ends, aseptate, and became dark brown at maturity, measuring 25.1 to 21.3 × 11.9 to 7.7 µm (n = 100 per isolate). These morphological characteristics were consistent with the genus Diplodia (Bhat et al. 2023), which may lead to canker, dieback, fruit rot, and leaf spots on economically important forest and horticultural species. For further molecular identification, the genomic DNA of three strains was extracted. The rDNA internal transcribed spacer (ITS) region and β-tubulin (tub) and translation elongation factor 1-alpha (tef1) genes were amplified using primers ITS1/ITS4, Bt2a/Bt2b, and EF1-728F/EF1-986R, respectively (Díaz et al. 2019). The ITS (PP957738 to PP957740), tub (PP960547 to PP960549), and tef1 (PP960550 to PP960552) sequences were submitted to GenBank. A phylogenetic tree of concatenated markers sequences (ITS, tub, and tef1) and ex-type strains sequences was constructed using MEGA software (version 11) with the neighbor-joining method. The three strains formed a clade with the strains CBS114791 and CBS113508 of D. seriata in the phylogenetic trees. Molecular analyses supported the identification of the strains as Diplodia seriata. To verify pathogenicity, conidial suspensions (106 conidia ml-1of isolate DS2) were sprayed onto the wound-free leaves of five A. japonica of three-month-old seedlings. In contrast, another five plants were sprayed with sterile water as a mock-inoculated control. Thirty days post-inoculation, lesions consistent with those observed in the field were present on the inoculated plants, while no symptoms appeared on the mock-inoculated controls. The entire inoculation experiments were repeated thrice. Isolates recovered from the symptomatic plants were identified as D. seriata based on morphological and ITS, tub, and tef1 sequence analyses, fulfilling Koch's postulates. Gummosis of Prunus persica and cankers on Juglans regia and Malus domestica caused by D. seriata have been reported in China (Wang et al. 2011; Zhang et al. 2017; Sha et al. 2023). This is the first report of D. seriata causing leaf blight on A. japonica. Leaf blight seriously affects the aesthetic landscape, and this finding provides valuable insights into the control and spread of D. seriata.

First report of Alternaria alternata causing postharvest fruit rot of sweet orange in Anhui province, China.

As an important cash crop, sweet orange (Citrus sinensis Osbeck cv. Newhall) is globally cherished by consumers for its health-promoting properties. March 2024, orange postharvest rot was found in three markets in Anqing, Anhui Province, China, with an incidence of 15% to 20%. The initial symptom was the appearance of small, light to dark brown spots on the surface of the fruit, which later expand and cause the fruit to rot. To isolate the pathogen, ten 5 × 5 mm pieces of symptomatic tissue from the infected oranges were surface sterilized in 1% Sodium hypochlorite for 60 s, rinsed three times with sterile water, and plated onto potato dextrose agar (PDA) at 25°C for 7 days. The pure culture was obtained by transferring single-spore isolates to fresh PDA medium. Ten isolates with similar morphological characteristics were isolated, and all of the colonies were dark brown with white margins and abundant aerial mycelium and sporulation. Conidia were pale to dark brown in chains or singly and were obclavate to obpyriform, measuring 5.6 to 28.0 × 4.7 to 16.3 μm (n = 30) with 1 to 5 transverse septa and 0 to 3 longitudinal septa. Their morphological characteristics were consistent with the description of Alternaria sp. (Simmons 2007). For molecular identification, we arbitrarily selected five isolates and named them CZ-1 to 5 for further investigation. The genomic DNA of CZ-1 to 5 was extracted, and the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF1) gene, and partial sequence of the β-tubulin (TUB2) gene were amplified with primers ITS1/ITS4, TEF1-F/TEF1-R (Zhang et al. 2020), and T1/T2 (O'Donnell and Cigelnik 1997), respectively. Sequences of the three loci from the representative isolate CZ-1 were deposited in GenBank (PP911430, PP928092, and PQ040398) and shared 100% nucleotide identity with Alternaria alternata sequences FJ717733, MH708309, and MN175551, respectively. A phylogenetic analysis was conducted using MEGA11 based on concatenated sequences of the three sequences, indicating that the isolates were closely clustered with reference strains of A. alternata. To evaluate pathogenicity, six surface-sterilized orange fruit with or without wounding were inoculated with a 20-μl drop of spore suspension (1 × 106 conidia/ml) of the isolates CZ-1 and -2. Another six fruit treated with sterile water with and without wounding were used as controls. All fruit were incubated in an artificial climate chamber at 25°C and 70 to 80% relative humidity for 7 days. Light to dark brown necrotic lesions at the wound sites appeared similar to those observed in the fruit markets, whereas control and nonwounded fruit remained healthy. The pathogens were re-isolated from the infected fruit, fulfilling Koch's postulates. The fungal pathogen A. alternata is widely distributed across various host plant species globally and has been documented as a causal agent of postharvest rot in kiwifruit (Li et al. 2017) and sweet cherry (Ahmad et al. 2020) in China. To our knowledge, this is the first report of A. alternata causing postharvest fruit rot of orange in Anhui province, China. The findings of this study will serve as a foundation for effectively managing the postharvest disease epidemic and prolonging the shelf life of oranges.

First Report of Leaf Spot Caused by Alternaria alternata on Artemisia stolonifera (Maxim) Komar. in China.

Artemisia stolonifera (Maxim.) Komar., also known as the 'elite of Artemisia', is currently widely planted in Jiangxi, Hubei, and other provinces in China. Since April 2023, a leaf spot on Ar. stolonifera was observed in the total cultivation area of 33.33 ha in Zhangshu City, Jiangxi Province. The incidence was 50% to 60%. Small irregular blackish-brown spots first appeared on the leaves, some with chlorotic margins, which later joined into clumps. In severe cases, the leaves were completely necrotic, seriously affected the growth and quality of Ar. stolonifera. Small pieces (3 to 4 mm) were excised from the necrotic borders of 20 typical symptomatic infected leaves, disinfected with 2.5% sodium hypochlorite solution for 3 min, rinsed five times with sterile water, and placed on potato dextrose agar (PDA) medium, incubated at 25℃ for 5 days. 15 purified isolates with similar morphological characteristics were obtained by transferring hyphal tips (isolation frequency of 75%). The colonies on PDA initially were white, and then became olive green with a white rim. After 10 days of culture on potato carrot agar medium (PCA), the conidia were septate, light brown in color, with dimensions ranging from 10.82 to 30.59 × 7.12 to 11.97 μm (n=50). The culture and morphological characteristics corresponded to Alternaria spp. (Simmons 2007), and the representative isolates JNC01 and JNC02 were used for further identification. To further identify the isolates, the RNA polymerase II second largest subunit (RPB2), internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Alternaria major allergen (Alt a1), translation elongation factor 1-alpha (TEF-α) and histone 3 (HIS3) were amplified using primers RPB2-5F2/RPB2-7CR, ITS1/ITS4, gpd1/gpd2, Alt a1-F/Alt a1-R , EF1-728F/EF1-986R and H3-1a/1b, respectively (Glass and Donaldson 1995; Lawrence et al. 2013; Berbee et al. 1999; Hong et al. 2005). The subsequent sequences were deposited in GenBank (accession nos. PP101993 and PQ458469; PP350760 and PQ45545; PP409573 and PQ474679; PP746506 and PQ474680; PP375820 and PQ458468; PQ001738 and PQ458467 for JNCO1 and JNC02 respectively. NCBI BLASTn sequence analysis revealed that all sequences, except for HIS3 had 100% homology with those of the Al. alternata strain CBS 612.72 (RPB2: KP124777.1, ITS: KP124308.1, GAPDH: KP124165.1, Alt a1: KP123861.1, TEF-α: KP125084.1), meanwhile, HIS3 had 100% identity with the Al. alternata strain MRY1 (MK210171.1). Phylogenetic trees, constructed using concatenated sequences based on ITS, GAPDH, Alt a1, TEF and RPB2 genes, as well as one built solely with HIS3, both placed JNC01 and JNC02 in the Al. alternata clade with high confidence. To test pathogenicity, about 5 mL of conidia suspension (1×106 conidia/mL) of JNC01 was sprayed on five 1-year-old healthy seedlings, while five seedlings were sprayed with sterile distilled water as a control. All plants were covered with plastic bags and placed in a greenhouse at 25 ± 2℃. After 7 days, brown spots, some with chlorotic margins, were observed, similar to those observed in the field. The controls remained symptom-free. The pathogens reisolated from the diseased tissue were again identified as Al. alternata by both morphological and molecular identification based on ITS and HIS3. This study is the first report of Al. alternata causing leaf spot on Ar. stolonifera in China and provides vital information on the pathogen for further diagnosis and management of the disease.

A new leaf spot caused by Alternaria alternata on Atractylodes lancea in Jiangsu, China.

Atractylodes lancea is an important Chinese herbal medicine, which is mainly produced in Jiangsu province, China. In June 2022, leaf spots symptom were observed on some A. lancea seedlingsgrowing in a Chinese herbal medicine resource garden of NanjingBotanical Garden, Jiangsu. Approximately 75% of 100 A. lancea seedlings suffered from the disease. Initially, gray to black spots appeared at the tip of the blade, then spread deep into the petiole, finally causing the blade to wither and fall. To isolate the pathogen, five diseased leaves were collected from five different seedlings. Leaf sections (3 to 4 mm) were excised from the margins between healthy and diseased tissues, surface sterilized in 75% alcohol for 30 s, then in 1.5% NaClO for 90 s, rinsed three times in sterilized distilled water, plated on potato dextrose agar (PDA) and incubated at 25℃in darkness. Pure cultures were obtained by monosporic isolation. Eighteen isolates were obtained, and 77.8% of isolates was identified as Alternaria spp. A representative isolate, CS4-1 was used for further investigation. The colony of CS4-1, growing on PDA was cotton-like and black to brown with gray-white aerial hyphae on their surfaces, and dark gray on the back. The conidia were solitary on conidiophores and were oval to pear-shaped, brown in color, with 1 to 4 transverse septa and 0 to 1 oblique septa, parietal cells extending into the beak, and measured 8.9 to 39.5×6.0 to 13.5 µm (n=35). These characteristics were consistent with the description of Alternaria spp. (Simmons 2007). Six DNA regions, i.e.,internal transcribed spacer (ITS),large subunit ribosomal RNA (LSU), small subunit ribosomal RNA (SSU), anonymous region OPA10-2, Alt a 1 major allergen (Alta1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH)and translation elongation factor 1-alpha (TEF1)with the respective GenBank Accession No. OP836052, OP836054, OP836051, OP851487, OP851488, OP851489, and OP851490, were amplified and sequenced with the primer pairs described by White et al. (1990) and Woudenberg et al. (2015). A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7, and CS4-1 clustered in the A. alternata clade. To test pathogenicity, 12 leaves, on three one-month-old A. lancea seedlings (four leaves from each seedling) were wounded with a sterile needle and inoculated with 20 μL of conidial suspension (1×106 spores/mL) on the left sides of leaves. The right sides of the leaves were inoculated with 20 µL sterile water and used as the control. All inoculated detached leaves and seedlings were covered with clear polyethylene bags to keep moisture and were incubated in a greenhouse at 25℃, 80% relative humidity, and a 12-h light/dark cycle. The experiment was repeated three times. After 4 days, typical gray to black spots were visible on the left sides of all inoculated leaves and the inoculated seedlings, and the right sides remained asymptomatic. Subsequently, the same fungus was reisolated and identified based on morphological and molecular traits. A. alternata has a very wide host range but has never been reported on A. lancea worldwide (nt.ars-grin.gov). Because of the medicinal value of A. lancea, further studies should be directed toward control of this disease.

Molecular phylogeny and genetic diversity of Fusarium oxysporum from various hosts in Malaysia

Fusarium oxysporum is a cosmopolitan fungus, consisting of both pathogenic and non-pathogenic members and known to be the causative agent of several diseases on various host plants. In Malaysia, most studies have focused on pathogenic F. oxysporum isolates because of their implications for agricultural production, but less attention has been given to non-pathogenic isolates. The aim of this study was to determine the phylogenetic relationship, genetic diversity, pathogenicity and host range of F. oxysporum in Malaysia. A total of 133 isolates of F. oxysporum were isolated from symptomatic plants of Abelmoschus esculentus, Solanum melongena, Solanum tuberosum, Cucumis sativus, Solanum lycopersicum, Cucumis melo, Musa paradisiaca var. awak, Hymenocallis littoralis, Asparagus officinalis, and Sansevieria trifasciata and non-agricultural soils in Malaysia. Comparison of nucleotide sequences of translation elongation factor 1-alpha (tef1-α) and mitochondrial small subunit (mtSSU) showed that the isolates were 98–100% similar to F. oxysporum from GenBank, thus, confirming the fungal identity. Besides, Malaysian isolates of F. oxysporum exhibited polyphyletic evolutionary origin, wide host range, and genetically diverse by grouping into 20 VCGs and 17 IGS haplotypes. This finding is beneficial for the purpose of quarantine, monitoring and disease management in the agricultural settings in Malaysia.