• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.393-399
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• 1998

Enterobacter sp.B54 which shows antagonistic activity to Phytophthora capsici on potato dextrose agar was selected among 112 strains isolated from Korean soil. After Tn5 lac-induced mutants were obtained through Pl :: Tn5 lac mutagenesis, 2 mutants for loss of antibiosis and 1 mutant for increased antibiosis were screened by using in vitro fungal inhibition assay. When the 3 mutants affected in antibiosis were analyzed by southern hybridization with pRZ102 (ColEl :: Tn5) as a probe, its results suggest that Tn5 lac was randomly inserted into different chromosomal sites in these mutants.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.598-608
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• 2019

Endophytic fungi have received much attention as plant growth promoters as well as biological control agents against many plant pathogens. In this study, 30 endophytic fungal species, isolated from various plants in China, were evaluated using in vitro dual culture assay against Fusarium oxysporum f. sp. cucumerinum, causing wilt in cucumber. The results of the present study clearly showed that all the 30 endophytic fungal isolates were highly capable of inhibiting the mycelial colony growth of Fusarium oxysporum f. sp. cucumerinum with inhibition % over 66% as compared to control treatments. Among all of them, 5 isolates were highly effective such as, Penicillium sp., Guignardia mangiferae, Hypocrea sp., Neurospora sp., Eupenicillium javanicum, and Lasiodiplodia theobromae, respectively. The Penicillium sp. and Hypocrea sp. were highly effective as compared to other isolates. From in vitro results 10 best isolates were selected for greenhouse studies. The results of the greenhouse studies showed that among all of them 3 endophytic fungal isolates successfully suppressed wilt severity when co-inoculation with pathogen Fusarium. oxysporum f. sp. cucumerinum. The endophytic fungi also enhanced plant growth parameters of the host plants, the antagonistic fungal isolates increased over all plant height, aerial fresh, and dry weight as compared to control.

• The Plant Pathology Journal
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• v.16 no.3
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• pp.125-129
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• 2000

A rapid and efficient assay to determine melanin biosynthesis inhibition of Magnaporthe grisea, a causal agent of the rice blast, by chemicals was developed. Wells in 24-well plates were loaded with spore suspension of the fungus and three known melanin biosynthesis inhibitors of KC10017, tricyclazole, and carpropamid. Subsequent color changes of mycelia and culture media in the wells were observed 7 days after incubation. The wells treated with KC10017 (an inhibitor of polyketide synthesis step and/or pentaketide cyclization step) became colorless, whereas tricyclazole (an inhibitor of 1, 3, 8-trihydroxynaphthalene reductase) or carpropamid (an inhibitor of scytalone dehydratase)-treated wells exhibited red color. They did not show any inhibitory effect on fungal growth. The inhibition of reaction steps prior to 1, 3, 6, 8-tetrahydroxynaphthalene formation was easily determined by colorless medium and mycelia. However, it was impossible to distinguish between inhibition of reduction steps and inhibition of dehydration steps by colors of the cultures. It was accomplished through HPLC analysis of the melanin biosynthesis-involving pentaketide metabolites accumulated by the inhibitors. Through screening of a number of synthetic chemicals using the in vitro assay, we could find a novel chemical group of melanin biosynthesis inhibitor.

• Mycobiology
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• v.48 no.4
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• pp.326-329
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• 2020

Valuable natural compounds produced by a variety of microorganisms can be used as lead molecules for development of new agrochemicals. Furthermore, high-throughput in vitro screening systems with specific modes of action can increase the probability of discovery of new fungicides. In the current study, a rapid assay tested with various microbes was developed to determine the degree of respiratory inhibition of Saccharomyces cerevisiae in two different liquid media, YG (containing a fermentable carbon source) and NFYG (containing a non-fermentable carbon source). Based on this system, we screened 100 fungal isolates that were classified into basidiomycetes, to find microbial secondary metabolites that act as respiratory inhibitors. Consequently, of the 100 fungal species tested, the culture broth of an IUM04881 isolate inhibited growth of S. cerevisiae in NFYG medium, but not in YG medium. The result is comparable to that from treatment with kresoxim-methyl used as a control, suggesting that the culture broth of IUM04881 isolate might contain active compounds showing the inhibition activity for respiratory chain. Based on the assay developed in this study and spectroscopic analysis, we isolated and identified an antifungal compound (-)-oudemansin A from culture broth of IUM04881 that is identified as Oudemansiella venosolamellata. This is the first report that (-)-oudemansin A is identified from O. venosolamellata in Korea. Taken together, the development of this assay will accelerate efforts to find and identify natural respiratory inhibitors from various microbes.

• Archives of Pharmacal Research
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• v.24 no.5
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• pp.397-401
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• 2001

In the search for new naturally occurring angiogenic inhibitory we found that culture broths from two unidentified fungal strains exerted potent inhibitory activities on capillary-like tube formation of human umbilical vein endothelial cells (HUVEC) in vitro. Two active compounds were isolated by bioassay-guided separation and their structures were identified as gliotoxin (1) and its derivative methylthiogliotoxin (2) by spectroscopic analyses. These compounds significantly inhibited the migration of HUVEC assessed by in vitro wounding migration assay and exhibited at least 10 times more potent inhibition of proliferation of HUVECs as compared with that of cancer cell lines such as HeLa, MCF-7, and KB 3-1 cells. Especially, gliotoxin having disulfide group exerted more potent activities than methylthiogliotoxin, suggesting that gliotoxin could be a useful compound for further study as an anti-angiogenic agent.

• Mycobiology
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• v.36 no.4
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• pp.260-265
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• 2008

Cyclic voltammetric measurements were performed for Co(II), Ni(II), Cu(II) and Zn(II) complexes of 1 : 1 alternating copolymer, poly(3-nitrobenzylidene-1-naphthylamine-co-succinic anhydride) (L) and Ni(II) and Cu(II) complexes of 1 : 1 alternating copolymer, poly(3-nitrobenzylidene-1-naphthylamine-co-methacrylic acid) ($L^1$). The in vitro biological screening effects of the investigated compounds were tested against the fungal species including Aspergillus niger, Rhizopus stolonifer, Aspergillus flavus, Rhizoctonia bataicola and Candida albicans and bacterial species including Staphylococcus aureus, Escherichia coli, Klebsiella pneumaniae, Proteus vulgaris and Pseudomonas aeruginosa by well diffusion method. A comparative study of inhibition values of the copolymers and their complexes indicates that the complexes exhibit higher antimicrobial activity. Copper ions are proven to be essential for the growth-inhibitor effect. The extent of inhibition appeared to be strongly dependent on the initial cell density and on the growth medium. The nuclease activity of the above metal complexes were assessed by gel electrophoresis assay and the results show that the copper complexes can cleave pUC18 DNA effectively in presence of hydrogen peroxide compared to other metal complexes. The degradation experiments using Rhodamine B dye indicate that the hydroxyl radical species are involved in the DNA cleavage reactions.

• YAKHAK HOEJI
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• v.42 no.5
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• pp.480-486
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• 1998

Many antitumor and immune modulating components have been isolated from fungal extracts. In this study, the authors isolated the proteoglycans from cultured mycelia of Lentin us lepideus, including especially the acidic polysaccharide fraction, named lepidan. It was obtained by extraction with hot water followed by purification using DEAE cellulose anion exchange. To elucidate antitumor effects against different type of tumor, the proteoglycans were tested on sarcoma 180, C3H MCA clone 16 and P388 leukemia in vivo. Lepidan showed 58.3% of tumor inhibition against solid form of sarcoma 180 and 58.6% against MCA clone 16. But lepidan did not affect life span of mice against P388 leukemia. Also when Lepidan was applicated to MTT assay, it did not show any direct cytotoxicity against various tumor cells in vitro.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.36-47
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• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.31-37
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• 2020

We previously identified a new heparinase inhibitor fungal metabolite, named CRM646-A, which showed inhibition of heparinase and telomerase activities in an in vitro enzyme assay and antimetastatic activity in a cell-based assay. In this study, we elucidated the mechanism by which CRM646-A rapidly induced nucleus condensation, plasma membrane disruption and morphological changes by increasing intracellular Ca²⁺ levels. Furthermore, PD98059, a mitogen-activated protein kinase (MEK) inhibitor, inhibited CRM646-A-induced nucleus condensation through ERK1/2 activation in rat 3Y1 fibroblast cells. We identified CRM646-A as a Ca²⁺ ionophore-like agent with a distinctly different chemical structure from that of previously reported Ca²⁺ ionophores. These results indicate that CRM646-A has the potential to be used as a new and effective antimetastatic drug.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.339-347
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• 2017

Sclerotium rolfsii causing southern blight on numerous vegetable and fruit crops was isolated from stems of chili peppers showing wilting symptoms. The pathogen was identified by morphological observation and DNA sequencing analysis of ITS region. To select an effective fungicide for control of southern blight, we investigated the inhibition efficacy of thirty fungicides included in nine groups of fungicides with different mechanisms of action. A fungal growth inhibition assay was conducted through an agar dilution method by using mycelial discs and sclerotia of the pathogen as inoculum, respectively. When mycelial discs were used as an inoculum, several fungicides showed good inhibitory activity against the mycelial growth of S. rolfsii 12-6. All DMI fungicides tested had a good inhibition except for prochloraz which had low inhibitory effect. All strobilurin fungicides tested except for kresoxim-methyl and all SDHI fungicides tested except for boscalid and fluopyram, had a good inhibition. Also, fludioxonil, a protective fungicide and fluazinam had a good inhibitory effect. Interestingly, when sclerotia were used as an inoculum, inhibition efficacy was increased for fluopyram, a SDHI fungicide, and for some protective fungicides such as propineb, chlorothalonil, dithianon, and folpet. All the fungicides selected in this study should be tested in the field for their control activities against stem rot for practical use in chili pepper cultivation.