Potato Dextrose Agar Research Articles

Identification and biological characteristics of Fusarium tobaccum sp. nov., a novel species causing tobacco root rot in Jilin Province, China.

Fusarium wilt of tobacco (FWT), caused by Fusarium spp., has emerged as a severe threat to tobacco production in China. In all, 132 isolates of Fusarium were isolated from tobacco and pathogenic to tobacco-causing FWT in Jilin Province, China. In this study, we identified 7 of 132 isolates as a novel species Fusarium tobaccum sp. nov. Zhao Xie & Jie Gao, using multi-gene phylogenetic analyses of translation elongation factor (tef1), β-tubulin (tub2), calmodulin (cmdA), and RNA polymerase II second largest subunit (rpb2) genes, along with subtle morphological differences. Isolates of F. tobaccum sp. nov. were clustered in a distinct branch in the maximum parsimony phylogenetic tree generated from the sequences of tef1-rpb2-tub2-cmdA and can be distinguished from closely related species F. cugenangense, F. callistephi, and F. elaeidis. The morphological characteristics of F. tobaccum sp. nov. are distinct from other Fusarium species. F. tobaccum sp. nov. exhibits abundant aerial mycelia and pigment production on potato dextrose agar (PDA), microconidia with 0-1 septa, and macroconidia with 2-5 septa on carnation leaf-piece agar (CLA), and produces abundant chlamydospores on Spezieller Nährstoffarmer agar (SNA) and CLA. The mycelia of F. tobaccum exhibited optimal growth at a pH of 7.1 and a temperature of 23.6°C. Sucrose and NaNO3 significantly promoted the mycelial growth of F. tobaccum. PD medium was optimal for total sporulation. However, the sporulation ratios of the macrospores of F. tobaccum in PD, SN, and CMC were relatively low (0.48%, 2.51%, and 2.16%, respectively). These findings provided valuable insights into the morphological and biological characteristics of F. tobaccum.IMPORTANCEFusarium wilt of tobacco (FWT) is a prevalent issue in tobacco-growing regions globally, leading to significant losses in yield and quality. This study identified F. tobaccum sp. nov., a novel species of Fusarium causing FWT in China. The identification was based on multi-gene phylogenetic analyses and morphological characteristics. The effects of temperature, pH, carbon source, nitrogen source, medium, and light on the mycelial growth of F. tobaccum sp. nov. were determined. These findings might contribute to future research on the pathogenic mechanisms of this novel species and the development of strategies to control FWT.

Feasibility of Fruit Waste-Derived Media for Microbial Culture: A Sustainable and Low-cost Approach

The present study, conducted at the College of Agriculture, Latur during 2023-2024, explores the viability of using fruit waste-derived media as a cost-effective alternative to conventional media for the growth of plant pathogens. The research focuses on the growth performance of Pseudomonas fluorescens, Fusarium oxysporum f. sp. udum, and Sclerotium rolfsii on media prepared from fruit peel of eight fruits (Banana, Pineapple, Papaya, Orange, Guava, Pomegranate, Dragon fruit, and Sapota). These fruit-based media were compared to traditional Nutrient Agar (NA) and Potato Dextrose Agar (PDA). Results revealed that Sclerotium rolfsii showed maximum growth on Papaya Dextrose Agar, while Pseudomonas fluorescens exhibited the most robust growth on Banana, Papaya and Dragon Fruit Dextrose Agar. Fusarium oxysporum f. sp. udum demonstrated the highest radial growth on Banana Dextrose Agar. The study also observed that fruit-based media significantly reduced costs compared to Potato Dextrose Agar and Nutrient Agar making them a sustainable and accessible option for microbiological research, particularly in resource-limited environments.

First Report of Root Rot Caused by Fusarium equiseti on Syrian rue (Peganum harmala) in China.

Syrian rue (Peganum harmala L) belongs to the family Zygophyllaceae and is mainly distributed in arid and semi-arid areas of Xinjiang, Gansu, Ningxia, Qinghai, and Inner Mongolia in China. It serves as a pioneer species in soil and water conservation, as well as in reclamation of wastelands, playing a crucial role in soil preservation and stabilization against sand encroachment. In 2023, a disease resembling root rot and wilt affected the quality and yield of Syrian rue in Xilingol (122.67°N, 42.78°E), Inner Mongolia, China. Symptoms ranged from yellowing to wilting, accompanied by darkening of the xylem in the roots. This disease was observed in approximately 50% of Syrian rue plants in desertification-prone grassland. A total number of 45 symptomatic root samples were collected from Syrian rue plants to isolate the pahogen. Small pieces of diseased tissue were surface sterilized with 75% alcohol for five seconds, rinsed twice with sterilized water, dried, and then placed on water agar and incubated for 72 h at 25℃ in darkness. A total of 45 isolates were obtained, showing identical morphology and sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-alpha (EF1-α) and RNA polymerase II second largest subunit (RPB2) genes. Therefore, the isolate LTP-5 was used as the representative for further study. On potato dextrose agar (PDA), fungal colonies of LTP-5 exhibited yellow to light brown colore with white aerial mycelia and irregular growth after 7 days of incubation at 25℃. Conidia and chlamydospre development were observed in carboxymethylcellulose sodium medium. Chlamydospores were abundant, terminal or intergrown between hyphae, rough brown walls of 5.9 to 19.82 μm in diameter (n = 75) were observed in the culture. Macroconidia had 5 to 7 septa, thick in the middle and thin at both ends, measured 28.17 to 62.81 μm in length and 1.50 to 4.19 μm in width (n = 128). Microconidia were absent. The morphological characteristics were consistent with the descriptions of Fusarium equiseti (Nelson et al., 1983). The pathogen was further confirmed to be F. equiseti by sequence analysis of the ITS, EF1-α and RPB2 genes amplified using polymerase chain reaction (PCR) with the primers ITS4/ITS5 (White et al., 1990), EF-1/EF-2 (O'Donnell et al., 1998), and RPB2-5F2 /fRPB2-7cR (Zhen et al., 2017). The sequences showed 100% similarity to the F. equiseti strain in the NCBI GenBank with accession numbers MH054914.1, KJ396323.1, and KT213286.1 for ITS, EF1-α, and RPB2, respectively. The sequences of PCR products were submitted to GenBank with accession numbers PP814863.1 (ITS), PP831628.1 (EF1-α) and PP826937.1 (RPB2). Pathogenicity tests were conducted through a pot assay. Five Syrian rue seedlings were inoculated by immersing plant roots in a suspension of 1×107 spores/mL for 30 min and transplanting to pots containing autoclaved soil. Control plants were immersed in sterile water. The inoculated plants showed yellow and root rot but no symptoms were observed on control plants. The same fungus was successfully re-isolated and identified based on the morphological characterization and sequence analyses from the infected root tissue but not from non-inoculated plants. In addition, F. equiseti has been reported to cause disease on Sugar beet (Khan et al., 2021), Watermelon (Rahman et al., 2021), potato (Cui et al., 2021), and other plants. This is the first report confirming F. equiseti as causal agent of Syrian rue wilt and root rot in China.

First Report in China of Leaf Blight on Spatholobus suberectus Dunn Caused by Diaporthe tulliensis.

Spatholobus suberectus Dunn is an important Chinese medicinal plant, which is the only officially recognized authentic source of the Chinese herbal medicine Spatholobi caulis in the China Pharmacopeia (Qin et al. 2024). In November 2023, a serious leaf disease was observed in the S. suberectus planting base in Nanning city (108.22E, 22.51N), Guangxi, China, with an incidence rate of 80%. Initially the symptoms appeared in the diseased leaves tips and was irregular in shape. The necrosis displayed discoloration of the gray color with a deep brown margin and this gradually expanded through into the lamina. To isolate the causal agent, 10 symptomatic leaves were collected and small segments (5×5 mm) of infected tissues were cut, and then they were surface sterilized (30s in 75% EtOH, 2 min in 2.5% NaOCl, and rinsed three times in sterilized distilled water), placed on potato dextrose agar (PDA) and incubated at 28℃ for 5 days. The hyphal tips were transferred to PDA to obtain pure cultures. 36 isolates were obtained, 23 of them had similar morphology, which were isolated from all the symptomatic leaves and were considered pathogens, and named YY-3-S. After 7 d, the colonies were white, fluffy, with white aerial mycelium. Sporulation was induced using pine needle medium (Su et al. 2012). After 30 days, the globose pycnidia developed, which were superficial to semi-immersed on the pine needle medium, and they appeared to be orange to black in color, with pale yellow conidial droplets. The alpha conidia spores were 3.53 to 6.85×1.75 to 3.70 μm (n = 50). They were unicellular, hyaline, aseptate and ellipsoidal to fusiform. These morphological features were consistent with Diaporthe sp (Sun et al. 2021). For further molecular identification, isolate YY-3-S underwent sequencing of the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1) sections (Huang et al. 2021). Obtained sequences of ITS, TUB2 and TEF1 (Genebank accessions nos. PP838571, PP865050 and PP865051) displayed a 99% similarity to Diaporthe tulliensis (Genebank accessions nos. OP163562, KR936132, OR662168, respectively). Phylogenetic analysis was based on a combination of ITS, TUB2 and TEF1 sequence data using MEGA 11 software to construct a phylogenetic tree with Neighbor-Joining. In the phylogenetic tree, the combined sequences attributed YY-3-S to D. tulliensis. The pathogenicity test was conducted on 1-year-old S. suberectus seedlings. We sprayed the mixed suspension of spores and mycelia on the unwounded leaves, while control seedlings received sterile water. 3 leaves per plant and 5 plants per treatment were selected for assessment. All seedings were placed in pots in a moist chamber at 28°C and 90% humidity. After 5 d, browning was observed on all the inoculated leaves tips. 14 d after inoculation, necrotic lesions expanded outward and their symptoms like those in the field, while control plants remained healthy. D. tulliensis was reisolated from the inoculated leaves, fulfilling the Koch's postulates. D. tulliensis has been reported to necrotic spots and leaf blight on Rambutan (Serrato-Diaz et al. 2023), leaf spot on bodhi tree (Li et al. 2022), stem canker on Jasmin (Hsu et al. 2023). To our knowledge, this is the first report of D. tulliensis causing leaf blight on S. suberectus in China. It is important to ensure timely management of the newly emerging disease.

First report of Curvularia chiangmaiensis causing leaf spot in elephant grass in Brazil.

Elephant grass, Cenchrus purpureus, exhibits high efficiency in sequesting atmospheric CO2 during photosynthesis, leading to significant biomass production. As a C4 plant, it holds immense potential for alternative biodiesel production and serves as fresh feed for cattle (Negawo et al. 2018). Field observations in commercial pastures planted with elephant grass, located in the Brazilian cerrado, revealed necrotic leaf spots with a brownish center and a yellowish halo, reaching a severity of 30 to 50% between 2022 and 2023 in Palmas (10°24'08"S 48°21'45" W) and Gurupi (11°46'21"S 49°02'08" W), municipalities located in the state of Tocantins, Brazil. In both years, the field incidence of the disease on young and mature leaves approached 100% during the rainy season. Affected leaves were collected, packed in a thermal box, and taken to the phytopathology laboratory. The 198 leaves were sanitised in 1% hypochlorite for 30 seconds and washed thrice in sterile water. Upon sanitisation, leaf tissues were cut and cultured on potato dextrose agar (PDA) medium and incubated in a controlled chamber (BOD) at 28°C with a 12-hour photoperiod. Colonies exhibited a greyish colour with whitish edges, and the top plate was brownish, growing to a diameter of 5 cm over seven days. The colonies obtained from monosporic cultures formed curved conidia, predominantly with three septa, of a brownish colour, with lengths ranging from (7 µm to 14 µm and widths from 5 µm to 7 µm. (N=-50, Fig 1) (Yasanthika et al. 2023). The UFTCC01 isolate's DNA was extracted using the extraction kit (PROMEGA ®), amplified for genes (ITS, Gapdh and Tefα1), sequenced (OR345316.1, OR344427.1 and OR344428.1) and compared in BLASTn at Gen Bank deposited sequences. Phylogenetic analysis using the parsimony method in MEGA X identified the isolate as Curvularia chiangmaiensis, showing 99.5% identity with reference sequences (MN215651.1, MN264085.1, and MN263942.1). Concatenated region comparison revealed 94% similarity to sequences (OP564987.1 and MF490814.1) in the phylogenetic tree, confirming the species. The pathogenicity test used 60 seedlings of 50-day-old elephant grass, 30 of which were inoculated with a spore suspension (3.0 × 106 spores/mL) and 30 with sterile water. The solutions were sprayed twice. Only plants inoculated with the spore suspension exhibited symptoms akin to those observed in the field. Then, the fungus was reisolated from the lesions and Koch's postulates were confirmed twice.Previous studies have documented the C. chiangmaiensis pathogenicity in other grasses, such as maise in Thailand (Marin et al. 2017). Our studies constitute the first report of C. chiangmaiensis causing leaf spots on elephant grass in the Brazil.

Comparison of rDNA-ITS Sequences, Vegetative Compatibility, Biological Characteristics, and Thifluzamide Sensitivity of Rhizoctonia solani AG-3 PT, TM, and TB Subgroup Isolates Collected from Potato and Tobacco in China.

Potato black scurf and tobacco target spot are significant agricultural diseases primarily caused by Rhizoctonia solani AG-3, which is further classified into PT (potato type), TM (tomato type), and TB (tobacco type) subgroups, with the naming of these subgroups originally indicating their respective infection hosts potato, tomato, and tobacco. This study determined the subgroup classification of 129 R. solani AG-3 isolates recovered from tobacco leaves exhibiting target spot disease in major tobacco-cultivating regions of China, as well as 80 R. solani AG-3 isolates obtained from potato stem cankers and tuber sclerotia in Liaoning, Jilin, and Heilongjiang provinces, and Inner Mongolia autonomous region of China. Sequence analysis of the internal transcribed spacer (ITS) regions revealed that all potato isolates belonged to the R. solani AG-3 PT subgroup, whereas the tobacco isolates were classified into TM and TB subgroups. Specifically, 51 strains from the northeast (Liaoning, Jilin, and Heilongjiang provinces) belonged to the TM subgroup, while the others belonged to the TB subgroup. Phylogenetic analysis indicated a closer relationship between AG-3 TM and AG-3 PT compared to AG-3 TB. Population genetic analysis highlighted distinct genetic variations among the subgroups, with the AG-3 TB subgroup isolates exhibiting fewer ITS1 variable sites and the AG-3 PT subgroup displaying a higher number of ITS2 variable sites. Furthermore, comparisons were made regarding macroscopic vegetative interactions, biological characteristics, pathogenicity to potato and tobacco, and susceptibility to thifluzamide among the three subgroups. The results indicated no macroscopic somatic interactions among the subgroups, with the highest frequency of somatic incompatibility observed within AG-3 PT, followed by AG-3 TM, and AG-3 TB. Additionally, the three subgroups differed in colony color, sclerotium morphology, production time, and distribution location on potato-dextrose-agar (PDA) medium, with AG-3 TM being the least likely to produce sclerotia. Regarding temperature, optimal growth conditions varied among the subgroups. Regarding pathogenicity, the AG-3 PT subgroup strains were more pathogenic on tobacco and potato stem bases compared to the AG-3 TB subgroup, whereas the AG-3 TB and TM subgroup strains were more pathogenic on leaves. Notably, AG-3 TM exhibited remarkable virulence towards both the stem base and leaves. The AG-3 PT subgroup strains exhibited the highest susceptibility to thifluzamide, with a susceptibility baseline of 0.037±0.013 mg·L-1, followed by the AG-3 TM subgroup strains with a baseline of 0.111±0.034 mg·L-1, and the AG-3 TB subgroup strains with a baseline of 0.137±0.035 mg·L-1. This study contributes to a deeper understanding of the biology and etiology of the three subgroups of R. solani AG-3, providing a valuable theoretical basis for the scientific control and management of tobacco target spot and potato black scurf diseases in China.

Evaluation of Antifungal Activity of Psidium guajava (Guava) Leaf Extracts

Aims: This study aimed to evaluate the effectiveness of four Psidium guajava leaf extracts against three strains of Candida albicans and a strain of Cryptococcus neoformans. Study Design: Experiment-observational study. Place and Duration of Study: P. guajava leaves were collected from specific areas in Regions Three and Four in Guyana; and the preparation of the leaf extracts and the treatment against the fungi were performed in two laboratories at the University of Guyana from August 2022-September 2023. Methodology: P. guajava leaves were washed with water, air-dried at room temperature, and ground in a disinfected food mixer. Extracts were prepared by soaking the ground leaves in different solvents, and then using the rotary evaporator to reduce the solutions and obtain crude extracts. Filter paper discs were prepared and allowed to soak in various concentrations of the crude extracts. The discs were then subjected to antifungal susceptibility testing on Potato Dextrose agar seeded with an appropriate fungus according to Clinical Laboratory Standards Institutes guidelines. The plates were incubated and the zone of inhibition (ZOIs) was measured. Results: P. guajava leaves were effective against all four strains of fungi evaluated. The largest ZOI was observed for C. neoformans (25 mm) with the ethyl acetate extract at 100 mg/mL concentration. Large ZOI was also observed for the three strains of C. albicans (10-15 mm). For one of the C. albicans strains, a ZOI significantly higher than the antifungal agents (ketoconazole and fluconazole) was observed with the P. guajava ethyl acetate extract at 100 mg/mL. Conclusion: It can be concluded that P. guajava leaves have antifungal activity against C. albicans and C. neoformans, potentially greater than current antifungal agents like ketoconazole and fluconazole.

Optimizing fungicide application timing for spring dead spot based on soil temperature and season

AbstractSpring dead spot (SDS) (Ophiosphaerella spp.) is the most detrimental disease to warm‐season turfgrasses in areas with cold‐induced dormancy. Fungicide applications do not provide consistent SDS suppression. One reason for this inconsistency is the use of solely calendar‐based fungicide applications instead of considering both calendar date and soil temperature. A field study was conducted at three separate hybrid bermudagrass (Cynodon dactylon (L.) Pers. × transvaalensis Burtt Davy) locations in Virginia to determine the optimal soil temperature and timing for SDS suppression with tebuconazole and isofetamid. Tebuconazole (1.5 kg a.i. ha−1) and isofetamid (4.1 kg a.i. ha−1) were applied at 11 different timings throughout the year based on soil temperatures at a 0‐ to 10‐cm depth. Plots were assessed for SDS severity in the spring and early summer of 2021 and 2022. Two in vitro studies were also conducted with Ophiosphaerella herpotricha and Ophiosphaerella korrae isolates to (1) determine the optimal temperature for growth on potato dextrose agar (PDA) placed on a thermogradient table (13–33°C) and (2) compare the daily growth rate of O. herpotricha and O. korrae isolates at 11, 19, and 27.5°C on PDA. In the field study, isofetamid suppressed SDS more than tebuconazole. Fall applications when soil temperatures were 13°C consistently provided the best SDS suppression. For the in vitro studies, both species grew optimally between 24 and 25°C, yet O. korrae and O. herpotricha growth rates differed at 11°C.

Optimized Liquid Medium Formulation for Sanghuangporus vaninii and Biological Activity of the Exopolysaccharides

Aims: Sanghuangporus vaninii (S. vaninii) is a rare medicinal mushroom that is rich in polysaccharides, triterpenes, flavonoids, and other bioactive compounds. It has good potential development value. Methods and Results: We performed single factor experiments and Box-Behnken response surface methodology to optimize the liquid fermentation medium formulation for S. vaninii with mycelial biomass as the indicator. The in vitro antioxidant and anti-cancer capacity of the exopolysaccharides of S. vaninii were estimated. The optimal liquid fermentation media composition for the MS-4, MS-6, and MS-8 strains of Sanghuangporus vaninii consisted of 25.86 ± 0.068 g/L maltose, 7.3 ± 0.043 g/L yeast extract, and 0.71 ± 0.005 g/L dandelion powder. The average mycelial biomass of S. vaninii under optimal conditions was 12.61 g/L. The mycelial biomass of the Sanghuangporus vaninii strains in the optimized formulation was 109–191% higher than that obtained with the basic potato dextrose broth (PDB). The exopolysaccharides of Sanghuangporus vaninii exhibited an ABTS radical scavenging activity with an EC50 of 0.021 ± 0.017 mg/mL and a DPPH radical scavenging activity with an EC50 of 0.076 ± 0.043 mg/mL. In anti-cancer assays, these exopolysaccharides demonstrated an IC50 value of 1.98 ± 0.36 mg/mL against PC-3 human prostate cancer cells, indicating significant bioactivity, highlighting their potential as functional food ingredients. Conclusions: In this study, the formula of liquid fermentation of S. vaninii strains was optimized, which lays a theoretical foundation for increasing the yield of S. vaninii and its application in industry. Moreover, our data showed the clinical potential of the S. vaninii exopolysaccharides as antioxidants and anti-cancer drugs.

First Report of Almond Decline Syndrome Caused by Neofusicoccum parvum in Italy

Almond (Prunus dulcis Mill. D. A. Webb) is considered a major crop in the Mediterranean countries. In Italy, almond orchards cover an area of 54,939 ha, with a yield of 82,146 t, with the cultivation widely distributed across several areas, such as southern regions. During the summer of 2023, 3-year-old almond plants cultivated in a commercial orchard in southern Piedmont (44°32’41.92’’ N; 7°57’25.7’’ E) showed wilting and sudden decline of the whole plant with an incidence of 15%. Main symptoms were cankers on the trunk characterized by a chocolate-brown discoloration of wood, cambium and even bark and occasional presence of gummy exudate. Symptomatic bark samples were rinsed with water, then blotted dry, and briefly soaked in ethanol for 15 s. Small pieces (5 × 5 mm) from the margin of necrotic areas were cut and placed on potato dextrose agar (PDA) plates amended with streptomycin sulphate (150 μg/ml). After incubation in the dark at 25 °C for 4 days fungal colonies emerging from the plated fragments were subcultured as monohyphal fungal cultures. They were fast-growing, turning from withish to smoky grey or grey-olivaceous after a week. Stromatic conidiomata were produced in pine needle agar, with pycnidia forming hyaline, globose to ellipsoidal, unicellular conidia, ranging in size from 16.9–17.3 μm × 5.4–5.6 μm. The causal agent was preliminary identified as Neofusicoccum sp. based on morphology. Two representative isolates (MTB3, MTC4) were identified by sequencing the ITS region and translation elongation factor 1-alpha (TEF1-α) gene with the primer pairs ITS1/ITS4 (White et al. 1990) and EF1-728F/EF1-986R (Carbone and Kohn 1999). The sequences of the two isolates were deposited in NCBI GenBank with accession numbers PP930646 / PP930647 (ITS), and PP933185 / PP933186 (TEF-1α). BLAST analysis revealed 99% similarity with reference sequences of Neofusicoccum parvum isolate CMW9081. A maximum-likelihood phylogenetic tree was constructed using MEGA7 software based on the combined ITS and TEF1-α dataset. The phylogenetic tree demonstrated that the two isolates grouped with N. parvum in a clade with the ex-type CMW9081. The pathogenicity of both isolates was evaluated on 1-year-old ‘Vairo’ almond potted plants grafted on GF677 rootstock, to assess Koch’s postulate. For the inoculation, 5 mm in diameter mycelium plugs taken from a 7-days-old PDA culture were placed on the surface of fresh wounds made with a sterile cork-borer (5 mm-diameter) on the stem. Control plants were inoculated with sterile PDA plugs. The test was conducted twice. Three blocks of three plants per isolate and control were arranged in a randomized block design. All inoculated plants showed symptoms of shoot blight and cankers 2-weeks after inoculation. Additionally, gumming, pycnidia production and internal wood discoloration developed on the inoculation sites, with lesion length spreading for 16.8 ± 1.6 and 15.4 ± 3.0 cm for MTB3 and MTC4, respectively. Most of the plants died one month after inoculation. Control plants showed no symptoms. The pathogen was consistently reisolated from inoculated plants and the identity was confirmed through TEF-1α sequencing. Thus, results of Koch postulate clearly identify N. parvum as the etiological agent of the observed disease of almond in Piedmont. N. parvum was recently reported in the same area on hazelnut (Waqas et al. 2021) and blueberry (Guarnaccia et al. 2021), and as causal agent of band canker and branch dieback in almond trees in US and Spain (Moral et al. 2019). To our knowledge, this is the first report of N. parvum as the causal agent of almond decline syndrome in Italy. Considering the increasing of almond cultivation in Italy, this disease represents a threat for the future production.

First Report of Southern Blight of Epimedium sagittatum Caused by Agroathelia rolfsii in Henan, China.

Epimedium [Epimedium sagittatum (Sieb. et Zucc.) Maxim., E. brevicornu Maxim., E. pubescens Maxim., E. koreanum Nakai] plants are perennial herbs and the dry leaves were used in traditional Chinese medicine with different medicinal effects (Chinese Pharmacopoeia, 2020 edition). In early August 2022 (the high temperature was 33 ℃ and the low temperature was 25 ℃ on average with above 70% relative humidity), large necrotized leaf spots were initially observed in one-year and two-year old E. sagittatum plants in Dengzhou (32°35'26.84" N, 112°11'8.37" E), Henan province, China. White mycelia, yellow and brown sclerotia were observed along the petioles and colonized to the back of leaves and also on soil-petioles interface around the diseased Epimedium plants. White mycelia spread rapidly from the disease plants as a center to neighboring plants and the infected plants eventually wilted. The disease incidence was 16.0% and 28.0% in the surveyed cultivation fields in Dengzhou (Henan) and Danjiangkou (Hubei), respectively, which resulted in above 20% of yield loss. The symptomatic tissues (petioles, n≥10) were cut into 0.5 cm sections with autoclaved scissors and surface sterilized with 75% ethanol for 30 s, followed by 3 min in 1% NaClO, rinsed three times with distilled sterile water, plated on autoclaved filter paper to remove excess moisture in a biological laminar hood, transferred onto potato dextrose agar (PDA) plates and incubated at 28℃ in incubator in the dark for 3-5 days. Twelve fungal isolates with white radial colony morphology were obtained and purified by hyphal-tip method. The isolates formed radial colonies showing abundant aerial mycelia with a growth rate of 19.25 to 22.25 mm/day (mean = 21.67 ± 0.77 mm; n = 36) and white abundant sclerotia were observed after 5-7 days post inoculation at 28℃. The hyphae of the isolates were hyaline, branched with clamp connections at septa. The isolates formed dark brown sclerotia on PDA plate post 10-14 days incubation. The diameter of mature brown sclerotia ranged from 1.31 to 3.07 mm (mean = 1.91 ± 0.39 mm; n = 40) and 13 to 45 sclerotia (mean = 28; n = 24) were observed on each Petri dish. The isolates were tentatively identified as Agroathelia rolfsii based on morphological characteristics (Yi Y., et al. 2024) and isolate YYHBJ1 was selected as representative isolate for molecular identification and pathogenicity test. The genomic DNA was extracted using CTAB method. For molecular identification, ITS region, translation elongation factor-1alpha (TEF-1α) and nuclear large-subunit ribosomal DNA (nLSU-rDNA) were amplified (White et al. 1990, Wendland and Kothe, 1997). The sequences 684-bp ITS (OR428367), 534-bp TEF1α (OR485567), and 965-bp nLSU-rDNA (OR426612) of isolate YYHBJ1 were deposited in GenBank. Sequence analysis revealed that ITS, TEF1α and nLSU-rDNA sequences of YYHBJ001 showed 100%, 99.63%, and 100% identity to ITS (MN380242 and JF819727), EF1α (MN262527) and nLSU-rDNA (MT225781 and AY635773) sequences of Agroathelia rolfsii (Sacc.) Redhead & Mullineux (Redhead and Mullineux 2023). Pathogenicity tests was confirmed by inoculating 1 year-old E. sagittatum plants grown in pots with sterile soil. PDA plug (5 mm diameter) colonized with 3 brown sclerotia and the fungal culture which was cultivated for 12 days was deposited beside the petiole on the surface of the soil. The control plants were inoculated with PDA plugs without the pathogen. Five plants were used in each treatment. The plants were cultivated in incubator at 28℃ with 80% relative humidity. Fourteen days later, white mycelia growing out from the sclerotia and spread along the petioles to the leave and the infected leaves were yellow. After 21 to 24 days, all inoculated plants displayed symptoms like those observed in field, while no symptoms were observed on the controls. The fungus was re-isolated from the inoculated plants as described above, fulfilling Koch's postulates. The pathogenicity tests were repeated three times. Based on morphological feature and DNA sequences, the pathogen of southern blight on Epimedium plants was identified as A. rolfsii (anamorph S. rolfsii). The pathogen A. rolfsii usually infect kinds of crops and cause southern blight, such as peanuts, chili peppers, green onion and so on (Daunde et al. 2018). To our knowledge, this is the first report of A. rolfsii causing southern blight on E. sagittatum plants in Henan in China.

First Report of Neofusicoccum nonquaesitum Causing Branch Dieback of English Walnut (Juglans regia) in the Maule Region, Chile

English walnut (Juglans regia) cultivation has been expanding in recent years in Chile, with a surface of 9,000 ha planted in the Maule Region (35° 26' S, 71° 40' W), central Chile. In a field survey conducted between August (2022) and January (2023) in three localities of the Maule Region, several declined trees were observed. English walnuts exhibited small chlorotic leaves, cankers, dead twigs, and dieback of branches, with an incidence of 10 to 35% of trees affected per orchard. Internally, diseased branches exhibit brown to dark brown wood cankers. Symptomatic branches (n = 30) were collected and surface disinfected with 96% ethanol, and flamed. Pieces of wood (approximately 5 mm) were placed on potato dextrose agar (PDA), supplemented with 0.005% tetracycline, 0.01% streptomycin, and 0.1% Igepal CO-630 (Díaz et al. 2018), and incubated at 22°C for 5 days in darkness. Four isolates (VLC-1-6-20, VLC-1-10-20, VLC-1-12-20, and VLC-1-15-20) exhibited moderate growth rates (16.4 mm/day), developing a white hue to olivaceous black colonies after 7 days at 22°C on PDA. Chlamydospores were absent. Black pycnidia were formed individually or in clusters. The isolates produced aseptate, hyaline, and fusiform, with base truncate conidia measuring 24.8 ± 1.6 x 6.8 ± 0.63 µm (l/w = 3.1; n = 50). The four isolates characterized were amplified using of the internal transcribed spacer (ITS1/ITS4), a portion of beta-tubulin (Bt2a/Bt2b), and part of the translation elongation factor 1- α (EF1-728/EF1-968) genes (Phillips et al. 2013). Blast analyses showed 100, 99 and 100 % identity with ex-type isolate PD-484 of Neofusicoccum nonquaesitum for ITS (deposited as GenBank accession no. PP697845 to PP697848), Bt (PP789576 to PP789579) and EF1 (PP830824 to PP830827) regions, respectively. Combined phylogenetic analysis using MEGA 7 software and the maximum likelihood test clustered the four isolates with ex-type of N. nonquaesitum. Pathogenicity was performed using two isolates (VLC-1-6-20 and VLC-1-10-20), which were inoculated in healthy tree walnuts cv. Chandler (8 years old), using 40 μl of mycelial suspension (105 fragments of mycelium/ml) on rounded wounds in the middle of each attached young branch (n=30 branches) (Twizeyimana et al. 2013). Sterile distilled water was used as a control treatment. Pathogenicity tests were repeated twice. After 6 months, necrotic streaks with mean lengths of 13 mm and 28 mm were observed in the middle of wounded young branches. No necrotic lesions were observed in the control treatments. Reisolation of N. nonquaesitum was positive (100%) only from inoculated branches and molecularly identified (EF1-a), fulfilling Koch’s postulates. Previously, N. nonquaesitum has been reported causing dieback on English walnut in California (Chen et al. 2014). To our knowledge, this is the first report of N. nonquaesitum causing canker and branch dieback of English walnut in central Chile. Previously, Diplodia mutila, Dothiorella sarmentorum, and N. austral have been associated with walnut dieback in Chile (Díaz et al. 2018; Iqbal et al. 2023; Barcos et al. 2023). In conclusion, it is imperative to implement epidemiological studies to avoid the spread and severity of branch dieback of walnuts in the Maule region, central Chile.

First Report of Leaf Spot Caused by Corynespora cassiicola on Pogostemon cablin in Guangdong, China.

Pogostemon cablin (Blanco) Benth. is a vital source of patchouli oil, utilized in traditional Chinese medicine, Ayurveda, cosmetics, and hygiene products(Swamy et al. 2016). In 2022, a leaf spot disease outbreak on patchouli plants occurred nearly 10 acres in Yunfu, Guangdong (21°2' N; 110°3' E), with an average incidence rate of 50%. Infected leaves initially showed circular spots with tan centers and yellow halos. Within five to ten days, these spots expanded, crossed leaf veins, and became polygonal with rough surfaces. As the disease progressed, the spots merged, darkening the veins and eventually causing leaf loss. Five symptomatic leaves were soaked in 75% ethanol for 10 s, and 2.5% sodium hypochlorite for 30 s, followed by a final rinse in sterile water. These samples were then placed on potato dextrose agar (PDA) plates. After 3-5 days of incubation at 28 °C, the mycelial growth was transferred to fresh PDA plates and purified by isolating the hyphal tip three times. Four of the five isolates had similar morphology and caused leaf spot symptoms upon inoculation. During culture on PDA, these isolates formed colonies with downy, gray-brown mycelium and darker centers. Microscopic examination revealed branched, septate mycelium; solitary, erect, unbranched grayish-brown conidiophores; and club-shaped conidia. Conidia were faint brown, segmented, contained 2 to 15 septa(predominant number being 3 to 4), ranged from 32 to 220 μm in length and 8.4 to 22.4 μm in width (n = 30). The morphology of these isolates was identical to that of Corynespora cassiicola (Ellis 1971). Pathogenicity tests were conducted on 3-month-old seedlings of P. cablin using the conidium infection method. A suspension of conidia (1 × 106 conidia/ml) was prepared from cultures induced to sporulate by 90-minute near-UV exposure followed by 2-day dark incubation at 28 °C, and 30 ml of this suspension was sprayed onto leaves of each seedling. Inoculated plants were incubated at 28 °C in an incubator. Potted plants treated only with sterile water were used as controls. Each treatment was inoculated into five potted plants. After seven days, all the inoculated leaves displayed symptoms similar to those observed in the fields, whereas the control leaves did not exhibit these symptoms. The pathogenicity test was repeated three times. Following Koch's postulates, the pathogen was re-isolated and identified as C. cassiicola through morphological and ITS sequence each time. To further identify, we selected a representative isolate, LD-TJ, for multi-locus sequence analysis of its ITS, LSU, and TEF1-α genes (GenBank Accession Nos. PQ042036, PQ035023, PQ060235)(Voglmayr et al. 2017). BLASTN analysis of the sequences obtained showed a high similarity of 99 to 100% with the ITS (JAEMHE010000031.1:51214-51777, 481/481 nucleotides), LSU (JAEMHE010000018.1:728464-729373, 910/910 nucleotides), and TEF1-α (JAEMHE010000005.1:656736-657712, 969/970 nucleotides) sequences of C. cassiicola CC01. The phylogenetic tree showed that the LD-TJ strain, C. cassiicola CC01(isolated from rubber trees), and C. cassiicola CC_29(isolated from soybean leaves) clustered into a clade with a 99% bootstrap value. C. cassiicola was identified as the cause of patchouli leaf spot in Hainan(Chen et al. 2010), but it has not yet been reported in Guangdong. Identifying P. cablin leaf spot disease is crucial in Guangdong Province because it is the main growing area for P. cablin in China (Yan et al. 2021).

Prevalence of Seed-Borne Fungi on Soybean (Glycine max L. Merr.) Seeds Stored Under Medium-Term Cold Room Facilities: Implications for Genebanks

This study examined the prevalence of seed-borne fungi in polyethylene plastic-packaged soybean seeds stored in a genebank and identified factors influencing their incidence. Twenty-four seed lots were sampled from the collection stored at 10 °C in the World Vegetable Center genebank in Arusha, Tanzania. The seeds used were those regenerated and harvested in 2015, 2016, and 2017. A seed health test was conducted for sterilized seeds on potato dextrose agar, sterilized seeds on top of the paper, and unsterilized seeds on top of the paper. Seven-year-old sterilized seeds plated on top of the paper exhibited the highest germination percentage (74 ± 7.09%) and lowest fungal incidence (7.00 ± 4.41%). Conversely, seven-year-old unsterilized seeds plated on top of the paper had the lowest germination (22.00 ± 6.97%) and highest fungal incidence (79.00 ± 8.31%). Older seed lots showed significantly higher equilibrium seed moisture content (Eqmc), though seed age had no significant influence on germination percentage or fungal incidence. Seed germination percentage showed a significant negative relationship with Eqmc, though Eqmc had no significant effect on fungal incidence. Meanwhile, germination percentage showed a significant negative relationship with fungal incidence. Six fungi isolates were identified using their morphological features from soybean seed samples: Absidia glauca, Aspergillus niger, Fusarium spp., Mucor hiemalis, Pestalotiopsis versicolor, and Sordaria macrospora. It is concluded that high fungal incidence negatively affects seed germination but is not significantly correlated with seed moisture content. The dry and cold storage conditions in the genebank, while effective in extending seed longevity, can unintentionally allow seed-borne mycoflora to persist. Although fungi may not actively proliferate under these conditions, they may do so when favorable conditions are restored, such as during germination tests, and consequently may compromise seed viability. Therefore, this study emphasizes the importance of increased systematic seed health checks before storage in genebanks to ensure optimal seed quality, and the replacement of all polyethylene plastic bags to aluminium foil packaging.

Utilization of Kluwih Seed Starch (Artocarpus camansi) as an Alternative Growth Medium for Aspergillus niger Fungus

Background: Carbohydrates in kluwih seeds have sufficient nutrients, such as potatoes and tubers, making it possible to be used as a medium for mushroom growth. The nutritional composition of breadfruit contains 25% carbohydrates, 1.5% protein, and 0.3% fat. Aspergillus niger fungi require nutrients such as carbohydrates and protein to grow. This study aims to determine whether kluwih seed starch can be used as an alternative medium for the growth of Aspergillus niger fungi. Method: This study used a pre-experimental design with laboratory observation. The study was conducted by utilizing kluwih seed starch as a medium, compared to Potato Dextrose Agar (PDA) and Sabouraud Dextrose Agar (SDA). Results: The results showed that kluwih seed starch can be used as an alternative medium for the growth of Aspergillus niger, with fungal colonies that grow macroscopically and microscopically almost the same as PDA and SDA media. Conclusion: Kluwih seed starch can be an alternative substitute for PDA media for the growth of Aspergillus niger.

Isolation, Antibacterial Activity and Molecular Identification of Avocado Rhizosphere Actinobacteria as Potential Biocontrol Agents of Xanthomonas sp.

Actinobacteria, especially the genus Streptomyces, have been shown to be potential biocontrol agents for phytopathogenic bacteria. Bacteria spot disease caused by Xanthomonas spp. may severely affect chili pepper (Capsicum annuum) crops with a subsequent decrease in productivity. Therefore, the objective of the study was to isolate rhizospheric actinobacteria from soil samples treated by physical methods and evaluate the inhibitory activity of the isolates over Xanthomonas. Initially, soil samples collected from avocado tree orchards were treated by dry heat air and microwave irradiation; thereafter, isolation was implemented. Then, antibacterial activity (AA) of isolates was evaluated by the double-layer agar method. Furthermore, the positive/negative effect on AA for selected isolates was evaluated on three culture media (potato-dextrose agar, PDA; yeast malt extract agar, YME; and oat agar, OA). Isolates were identified by 16S rRNA sequence analysis. A total of 198 isolates were obtained; 76 (series BVEZ) correspond to samples treated by dry heat and 122 strains (series BVEZMW) were isolated from samples irradiated with microwaves. A total of 19 dry heat and 25 microwave-irradiated isolates showed AA with inhibition zones (IZ, diameter in mm) ranging from 12.7 to 82.3 mm and from 11.4 to 55.4 mm, respectively. An increment for the AA was registered for isolates cultured on PDA and YME, with an IZ from 21.1 to 80.2 mm and 14.1 to 69.6 mm, respectively. A lower AA was detected when isolates were cultured on OA media (15.0 to 38.1 mm). Based on the 16S rRNA gene sequencing analysis, the actinobacteria belong to the Streptomyces (6) and Amycolatopsis (2) genera. Therefore, the study showed that microwave irradiation is a suitable method to increase the isolation of soil bacteria with AA against Xanthomonas sp. In addition, Streptomyces sp. BVEZ 50 was the isolate with the highest IZ (80.2 mm).

First report of leaf spot disease caused by Fusarium asiaticum on Lonicera caerulea L. in Heilongjiang Province, China.

Heilongjiang is the largest province with the small, cultivated fruit plant, blue honeysuckle (Lonicera caerulea L.), in China (Sun et al. 2024). Between September 2022 and September 2023, a leaf spot disease was observed on about 10 percent of blue honeysuckles of the 'Lanjingling' cultivar grown in a 6.66-ha field in Mudanjiang (129.38°E, 44.59°N), Heilongjiang Province, China. The typical symptoms include a few small brown sunken leaf spots (approximately 2 mm) in the fruiting stage that gradually expand into larger brown sunken lesions (approximately 10 mm) in the senescence stage (Fig. S1). Small 3-4-mm segments of infected tissue from 100 randomly selected leaf samples with typical symptoms were surface sterilized with 75% ethanol and 5% sodium hypochlorite, rinsed in sterile distilled water, and transferred to 9-cm Petri dishes containing potato dextrose agar (PDA) after drying. Finally, 10 isolated fungi were obtained through single-spore culture on PDA, and their conidia displayed a morphology consistent with Fusarium spp. (Zheng et al. 2023), and no other fungi were isolated. The fungal colonies on PDA were pink to dark red with fluffy aerial mycelium and pigmentation (Fig. S2). Microconidia were not observed, and the macroconidia of fungi had 2 to 6 septa, which were slender, sickle-shaped to nearly curved, with an average size of 38.83 (21.30 to 55.20) × 3.87 (2.70 to 6.60) μm (n = 50) on carnation leaf agar (Fig. S3). Genomic DNA was extracted from the representative isolate LD-245 for molecular verification, and PCR amplification was performed with ITS1/ITS4 (White et al. 1990), RPB2-5F2/RPB2-7CR (Liu et al. 1999), and EFl/EF2 (O'Donnell et al. 1998) primers. Sequences of LD-245 ITS (OR565903), RPB2 (OR584061), and TEF (PP965657) revealed 99 to 100% (494/494, 722/726, and 654/654) of identity with Fusarium asiaticum (MT322117, MH582121, and MT330257). Phylogenetic tree was constructed based on the concatenated sequences of ITS, RPB2, and TEF genes, and LD-245 was clustered with F. asiaticum (Fig. S4, Table S1). Therefore, based on the morphological characteristics and molecular phylogeny, LD-245 was identified as F. asiaticum. Six two-year-old healthy plants of the 'Lanjingling' cultivar were selected for a pathogenicity test. Three plants were inoculated with LD-245 conidial suspension (1 × 106 spores/ml) or with sterile water as an experimental control. All plants were cultured in a greenhouse (28℃, 75% relative humidity, 12 h light and dark cycle), and each experiment was replicated three times. Typical leaf spot symptoms were observed on inoculated leaves after 10 days (Fig. S5), whereas no symptoms were detected on water-treated leaves. The pathogens re-isolated from infected leaves displayed the same morphological and molecular traits as the initial isolate, and were again identified as F. asiaticum, confirming Koch's postulate. F. asiaticum was previously reported on giant elephant's ear (Alocasia macrorrhizos) (Zheng et al. 2023), causing leaf spot disease in China. This study is the first report of blue honeysuckle leaf spot disease caused by F. asiaticum in China. China has the largest blue honeysuckle cultivation area globally, and the production of blue honeysuckle with horticultural value is important for growers (Sun et al. 2024). Therefore, we hope researchers can develop efficient measures to manage emerging plant diseases in the future.

First report of leaf blight of water chestnut (Trapa natans L.) caused by Sclerotium hydrophilum in central China.

Trapa natans L., or water chestnut, is a currently globally distributed aquatic plant. The Yangtze River basin in China, a site origin of water chestnut (Fan et al., 2022), has extensive cultivation as a vegetable. In June 2021, a survey at the National Aquatic Vegetable Resource Garden in Wuhan, Hubei, China, revealed browning and wilting of water chestnut plants, with abundant white mycelia and brown to black sclerotia on leaves, indicative of southern blight (Sclerotium rolfsii). Field disease incidence a 1 ha area reached 70%, reducing fruit yield by 50%. About 10% of diseased plants showed blackening and rot from petioles to leaves. White sclerotial primordia and small brown to black sclerotia formed on the plant surfaces. By late August, symptoms exceeded 60%. To identify the pathogen, isolations were made from 243 sclerotial samples using 75% alcohol to disinfect, rinsed three times with water, and then incubated on potato dextrose agar (PDA) at 25℃. A total of 129 isolates were obtained, most of which exhibited characteristics of S. rolfsii. However, 21 isolates had sclerotia significantly smaller than those of S. rolfsii (1 to 1.5 mm in diameter). These isolates, cultured on PDA, produced abundant fluffy white aerial hyphae, 3 to 6 μm wide. Optimal mycelium growth was between 25 °C to 30 °C, with an average daily rate of 10 mm. White to light brown sclerotia appeared after 5 days and turned black within 10 to 14 days, averaging 0.34 mm in diameter (n=50). Some isolates produced a light brown pigment. These traits matched the description of S. hydrophilum (Bashyal et al. 2021). Isolates 221 and 238 were selected for molecular identification, with genomic DNA extracted from mycelia using the CTAB method. PCR amplification was conducted using ITS1/ITS4 and NS1/NS6 primers to target the internal transcribed spacer (ITS) and the small subunit ribosomal RNA gene (ssrRNA). Sequence analysis showed that the ITS sequence of isolate 221 (GenBank Acc. No. OR512512) had 99.84% sequence identity with S. hydrophilum Msh6 (GenBank Acc. No. FJ595946), and isolate 238 (GenBank Acc. No. PP035993) had 99.72% identity with S. hydrophilum Whcc-4 (GenBank Acc. No. PP035994). The ssrRNA sequences of both isolates (GenBank Acc. No. PP237261) had 99.69% identity with S. hydrophilum strain Hbq001 (GenBank Acc. No. KY995575), confirming their identification as S. hydrophilum. To assess pathogenicity, 16 water chestnut plants (cultivar Jia-yu Ling) at the rosette stage were placed individually in 32 cm diameter, 10 cm deep containers with fresh water. Eight plants were inoculated with 50 mature sclerotia from PDA cultures of isolates 221 and 238, and incubated at 25 °C for 14 days, with four plants per isolate. The remaining eight plants served as controls. Containers were covered to maintain 100% relative humidity at 25 °C to 32 °C for 3 days. This procedure was repeated twice. After 7 days, inoculated plants developed dark brown lesions on petioles that spread to leaves by 15 days post-inoculation. Fungi isolated from diseased leaves resembled isolates 221 and 238, fulfilling Koch's postulates. S. hydrophilum infecting at least 19 genera of plants, including rice (Zhong et al. 2018), wild rice, water lily (Kernkamp et al. 1977) and watershield (Fu et al. 2024). This is the first report of S. hydrophilum infecting water chestnut (T. natans) in central China, identifying it as a co-pathogen with S. rolfsii. Their combined presence may heighten plant mortality and threaten water chestnut cultivation.

Optimization of fermentation conditions for physcion production of Aspergillus chevalieri BYST01 by response surface methodology.

This study aimed to optimize the culture conditions of the termite-derived fungus Aspergillus chevalieri BYST01 for the production of physcion, a characteristic component of the traditional herb rhubarb, which has been commercially approved as a botanical fungicide in China. First, potato dextrose broth was screened as the suitable basal medium for further optimization, with an initial yield of 28.0 mg/L. Then, the suitable carbon source, fermentation time, temperature, pH value, and the rotary shaker speed for physcion production were determined using the one-variable-at-a-time method. Based on the results of single factors experiments, the variables with statistically significant effects on physcion production were further confirmed using the Plackett-Burman design (PBD). Among the five variables, temperature, initial pH, and rotary shaker speed were identified as significant factors (P < 0.05) for physcion productivity in the PDB and were further analyzed by response surface methodology (RSM). Finally, we found that the maximum physcion production (82.0 mg/L) was achieved under the following optimized conditions:initial pH 6.6, rotary shaker speed of 177 rpm, temperature of 28 °C, and glucose concentration of 30 g/L in PDB medium after 11 d of fermentation. The yield of physcion under the optimized culture conditions was approximately threefold higher than that obtained using the basal culture medium. Furthermore, the optimum fermentation conditions in the 5-L bioreactor achieved a maximal physcion yield of 85.2 mg/L within 8 d of fermentation. Hence, response surface methodology proved to be a powerful tool for optimizing physcion production by A. chevalieri BYST01. This study may be helpful in promoting the application of physcion produced by A. chevalieri BYST01 to manage plant diseases.

Fusarium verticillioides pigment: production, response surface optimization, gamma irradiation and encapsulation studies

BackgroundNatural pigments are becoming more significant because of the rising cost of raw materials, pollution, and the complexity of synthetic pigments. Compared to synthetic pigments, natural pigments exhibit antimicrobial properties and is less allergic. Pigments from microbial sources could easily be obtained in an inexpensive culture media, produced in high yields, and microbes are capable of producing different colored pigments. Searching for new sources for natural pigments to replace synthetic ones in food applications has become an urgent necessity, but the instability of these compounds is sometimes considered one of the obstacles that reduce their application. Encapsulation provides an ideal solution for natural dye protection through a controlled release strategy. Thus, this study aims at isolation of several soil fungi and subsequent screening their pigment production ability. The chosen pigment-producing fungal strain underwent full identification. The produced pigment was extracted with ethyl acetate and estimated spectrophotometrically. As there is a necessity to obtain a high pigment yield for efficient industrial application, the best production medium was tested, optimum conditions for maximum dye production were also investigated through the response surface methodology, and gamma irradiation was also employed to enhance the fungal productivity. Encapsulation of the produced pigment into chitosan microsphere was tested. The pigment release under different pH conditions was also investigated.ResultsA new strain, Fusarium verticillioides AUMC 15934 was chosen and identified for a violet pigment production process. Out of four different media studied, the tested strain grew well on potato dextrose broth medium. Optimum conditions are initial medium pH 8, 25 °C-incubation temperature, and for 15-day incubation period under shaking state. Moreover, a 400 Gy irradiation dose enhanced the pigment production. Chitosan microsphere loaded by the pigment was successfully prepared and characterized by infrared spectroscopy and scanning electron microscopy.ConclusionThis irradiated Fusarium strain provides a more economically favorable source for production of a natural violet dye with an optimum productivity, enhanced yield, and improved properties (such as, enhanced stability, controlled release, and bioaccessibility) by encapsulation with chitosan for efficient application in food industry.