• Title/Summary/Keyword: conidial germination

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Biocontrol Activity of Volatile-Producing Bacillus megaterium and Pseudomonas protegens against Aspergillus flavus and Aflatoxin Production on Stored Rice Grains

  • Mannaa, Mohamed;Oh, Ji Yeon;Kim, Ki Deok
    • Mycobiology
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    • v.45 no.3
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    • pp.213-219
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    • 2017
  • In our previous study, three bacterial strains, Bacillus megaterium KU143, Microbacterium testaceum KU313, and Pseudomonas protegens AS15, were selected as effective biocontrol agents against Aspergillus flavus on stored rice grains. In this study, we evaluated the inhibitory effects of the volatiles produced by the strains on A. flavus growth and aflatoxin production on stored rice grains. The three strains significantly reduced mycelial growth of A. flavus in dual-culture assays compared with the negative control strain, Sphingomonas aquatilis KU408, and an untreated control. Of these tested strains, volatiles produced by B. megaterium KU143 and P. protegens AS15 markedly inhibited mycelial growth, sporulation, and conidial germination of A. flavus on agar medium and suppressed the fungal populations in rice grains. Moreover, volatiles produced by these two strains significantly reduced aflatoxin production in the rice grains by A. flavus. To our knowledge, this is the first report of the suppression of A. flavus aflatoxin production in rice grains using B. megaterium and P. protegens volatiles.

Antagonistic Evaluation of Chromobacterium sp. JH7 for Biological Control of Ginseng Root Rot Caused by Cylindrocarpon destructans

  • Han, Joon-Hee;Park, Gi-Chang;Kim, Kyoung Su
    • Mycobiology
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    • v.45 no.4
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    • pp.370-378
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    • 2017
  • Cylindrocarpon destructans is an ascomycete soil-borne pathogen that causes ginseng root rot. To identify effective biocontrol agents, we isolated several bacteria from ginseng cultivation soil and evaluated their antifungal activity. Among the isolated bacteria, one isolate (named JH7) was selected for its high antibiotic activity and was further examined for antagonism against fungal pathogens. Strain JH7 was identified as a Chromobacterium sp. using phylogenetic analysis based on 16S rRNA gene sequences. This strain was shown to produce antimicrobial molecules, including chitinases and proteases, but not cellulases. Additionally, the ability of JH7 to produce siderophore and solubilize insoluble phosphate supports its antagonistic and beneficial traits for plant growth. The JH7 strain suppressed the conidiation, conidial germination, and chlamydospore formation of C. destructans. Furthermore, the JH7 strain inhibited other plant pathogenic fungi. Thus, it provides a basis for developing a biocontrol agent for ginseng cultivation.

Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum

  • Li, Taiying;Jung, Boknam;Park, Sook-Young;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.393-405
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    • 2019
  • Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

The Autophagy Protein CsATG8 is Involved in Asexual Development and Virulence in the Pepper Anthracnose Fungus Colletotrichum scovillei

  • Kwang Ho Lee;Adiyantara Gumilang;Teng Fu;Sung Wook Kang;Kyoung Su Kim
    • Mycobiology
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    • v.50 no.6
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    • pp.467-474
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    • 2022
  • Autophagy serves as a survival mechanism and plays important role in nutrient recycling under conditions of starvation, nutrient storage, ad differentiation of plant pathogenic fungi. However, autophagy-related genes have not been investigated in Colletotrichum scovillei, a causal agent of pepper fruit anthracnose disease. ATG8 is involved in autophagosome formation and is considered a marker of autophagy. Therefore, we generated an ATG8 deletion mutant, ΔCsatg8, via homologous recombination to determine the functional roles of CsATG8 in the development and virulence of C. scovillei. Compared with the wild-type, the deletion mutant ΔCsatg8 exhibited a severe reduction in conidiation. Conidia produced by ΔCsatg8 were defective in survival, conidial germination, and appressorium formation. Moreover, conidia of ΔCsatg8 showed reduced lipid amount and PTS1 selectivity. A virulence assay showed that anthracnose development on pepper fruits was reduced in ΔCsatg8. Taken together, our results suggest that CsATG8 plays various roles in conidium production and associated development, and virulence in C. scovillei.

Genetic Stability of Magnaporthe oryzae during Successive Passages through Rice Plants and on Artificial Medium

  • Park, Sook-Young;Chi, Myoung-Hwan;Milgroom, Michael G.;Kim, Hyo-Jung;Han, Seong-Sook;Kang, Seog-Chan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.313-320
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    • 2010
  • Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

Molecular characterization of yeast Snf1 homologue (sucrose non-fermenting gene) from Magnaporthe grisea

  • Yi, Mi-Hwa;Jeong, Jae-Gyu;Kim, Bong-Gyu;Ahn, Joong-Hoon;Lee, Yong-Hwan
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.84.2-85
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    • 2003
  • Magnaporthee grisea causes the devastating blast disease of rice. Entensive research has been conducted on infection mechanisms, particularly on appressorium formation and penetration, of this fungus during the last decade. However, the role(s) of cell-wall-degrading enzymes (CWDEs) on pathogenesis is not clearly demonstrated at molecular level. Many CWDES in plant pathogenic fungi including M. grisea are redundant; that is, there are multiple genes encoding enzymes with a similar or overlapping spectrum of activities. It is laborious to isolate all of the genes encoding related enzymes and to construct mutants lacking all 9f them. Thus, we considered alternative strategies to address the role of CWDEs in pathogenesis. Since expression of CWDE genes Is repressed by a simple sugar, as the first step, we cloned a Snfl (sucrose non-fermenting) gene (MgSnf1) from M. grisea. The predicted amino acid sequence showed a high identity with other Snf1 genes from various fungi. To elucidate molecular function of MgSnf1, a transformant lacking MgSnf1 was created by targeted gene replacement. En glucose, sucrose, and xylan the MgSnf1 mutant grew normally but in pectin and complex media, it grew slower than wild type. Expression of various CWDEs in MgSnf1 mutant was investigated and found that expression of some CWDEs is repressed. However, no significant difference was observed in conidial germination, appressorium formation, and pathogenicity in MgSnf1 mutant. However, MgSnf1 functionally complemented a yeast MgSnf1 mutant. These results suggest that MgSnf1 is involved in regulation of CWDEs and MgSnf1 is dispensable in pathogenicity of M. grisea.

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Biological Control of Gray Mold Rot of Perilla Caused by Boftis cinerea 1. Resistance of Perilla Cultivars and Selection of Antagonistic Bacteria

  • Moon, Byung-Ju;Son, Yeong-Jun;Lee, Jae-Pil;Kim, Choul-Seung;Song, Ju-Hee;Kim, Hyun-Ju;Kim, Jae-Woo;Kim, Do-Hoon;Park, Hyean-Cheal
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.36-42
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    • 2002
  • Resistance of perilla varieties to Botrytis cinerea LVF12 was evaluated, while antagonistic bacteria were selected and tested for their efficacy towards biological control of gray mold rot caused by B. cinerea. Among 11 perilla varieties tested for disease resistance, Milyang variety showed some degree of resistance, while the rest of varieties showed no resistance. Among 250 bacterial isolates collected from perilla loaves and rhizosphere of perilla plants, six isolates showed high levels of inhibitory effect on mycelial growth and conidial germination of B. cinerea in in vitro test. Using the pot test in growth chambers these isolates showed high levels of disease suppression, with Nl isolate showing 95.3% of control value and N4 isolate showing 90.8% of control value. Further test was performed to evaluate the two isolates ability for disease prevention and/or disease therapy, and results showed almost 100% of control vague. Isolates Nl and N4 were identified as Bacillus licheniformis and 5. megatepium, respectively, according to Bergey's manual, API 20E and 50CHB test kit, and Transmission electron microscope.

Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Anthocyanins from Clitoria ternatea Attenuate Food-Borne Penicillium expansum and its Potential Application as Food Biopreservative

  • Leong, Chean-Ring;Azizi, Muhammad Afif Kamarul;Taher, Md Abu;Wahidin, Suzana;Lee, Kok-Chang;Tan, Wen-Nee;Tong, Woei-Yenn
    • Natural Product Sciences
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    • v.23 no.2
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    • pp.125-131
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    • 2017
  • Clitoria ternatea or Commonly known blue pea, is a perennial climber crop native to Asian countries. The current study was aimed to evaluate the antimicrobial activity C. ternatea extract on food borne microorganisms and its antifungal effect on Penicillium expansum. The extract showed significant antimicrobial activity against 3 Gram positive bacteria, 2 Gram negative bacteria and 1 filamentous fungus on disc diffusion assay. The extract also showed good biocidal effect on all Gram positive bacteria tested and P. expansum. However, the kill curve analysis revealed that the fungicidal activity of the extract against P. expansum conidia was depend on the concentration of the extract and the time of exposure of the conidia to the extract. The scanning electron micrograph of the extract treated P. expansum culture showed alterations in the morphology of fungal hyphae. The germination of P. expansum conidia was completely inhibited and conidial development was totally suppressed by the extract, suggesting the possible mode of action of anthocyanin. Besides, the extract also exhibited 5.0-log suppression of microbial growth relative to control in the rice model. The results indicate the potential use of the C. ternatea anthocyanin as food biopreservative.

Biological Control with Streptomyces sp. on Fusarium oxysporum f. sp. vasinfectum and Phytophthora nicotianae var. parasitica Causing Sesame Wilt and Blight (Streptomyces sp. 에 의한 참깨 시들음병 (Fusarium oxysporum f. sp. vasinfectum) 및 역병 (Phytophthora nicotianae var. parasitica)의 생물학적(生物學的) 방제(防除))

  • Chung, Bong-Koo;Hong, Ki-Sung
    • The Korean Journal of Mycology
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    • v.19 no.3
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    • pp.231-237
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    • 1991
  • This study was conducted in order to find out biological control of sesame wilt and blight caused by Fusarium of oxysporum f. sp. vasinfectum and Phytophthora nicotianae var. parasitica by using Streptomyces spp. Two sesame pathogens, Fusarium oxysporum f. sp. vasinfectum and Phytophthora nicotianae var. parasitica were purely isolated from diseased sesame plants of the field. Streptomyces species were isolated from 72 soil samples collected from red pepper and sesame uplands in Chungbuk and selected as antagonists according to the results of dual culture. The selected Streptomyces isolates such as St-11 and St-20 were confirmed their antagonistic effect through mycelial inhibition zone and inhibitory effects on the mycelial growth of the pathogens by culture filterate of the antagonists. Inhibitory effects on the conidial germination of Fusarium oxysporum vasinfectum and Phytophthora nicotianae parasitica by the antagonists were also tested in addition to mycelial Iysis. The antagonists St-11 and St-20 showed inhibitory effect on growth of sesame seedlings after seeds soaked in the suspension. Effect of soil inoculation with antagonist St-11 showed 40 to 78 percent of control effect for two diseases in comparison with control under greenhouse.

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