• The Plant Pathology Journal
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• v.38 no.4
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• pp.313-322
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• 2022

Seed-borne pathogens in crops reduce the seed germination rate and hamper seedling growth, leading to significant yield loss. Due to the growing concerns about environmental damage and the development of resistance to agrochemicals among pathogen populations, there is a strong demand for eco-friendly alternatives to synthetic chemicals in agriculture. It has been well established during the last few decades that plant seeds harbor diverse microbes, some of which are vertically transmitted and important for plant health and productivity. In this study, we isolated culturable endophytic bacteria and fungi from soybean seeds and evaluated their antagonistic activities against common bacterial and fungal seed-borne pathogens of soybean. A total of 87 bacterial isolates and 66 fungal isolates were obtained. Sequencing of 16S rDNA and internal transcribed spacer amplicon showed that these isolates correspond to 30 and 15 different species of bacteria and fungi, respectively. Our antibacterial and antifungal activity assay showed that four fungal species and nine bacterial species have the potential to suppress the growth of at least one seed-borne pathogen tested in the study. Among them, Pseudomonas koreensis appears to have strong antagonistic activities across all the pathogens. Our collection of soybean seed endophytes would be a valuable resource not only for studying biology and ecology of seed endophytes but also for practical deployment of seed endophytes toward crop protection.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.315-324
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• 2016

Background: Biocontrol agents are regarded as promising and environmental friendly approaches as agrochemicals for phytodiseases that cause serious environmental and health problems. Trichoderma species have been widely used in suppression of soil-borne pathogens. In this study, an endophytic fungus, Trichoderma gamsii YIM PH30019, from healthy Panax notoginseng root was investigated for its biocontrol potential. Methods: In vitro detached healthy roots, and pot and field experiments were used to investigate the pathogenicity and biocontrol efficacy of T. gamsii YIM PH30019 to the host plant. The antagonistic mechanisms against test phytopathogens were analyzed using dual culture, scanning electron microscopy, and volatile organic compounds (VOCs). Tolerance to chemical fertilizers was also tested in a series of concentrations. Results: The results indicated that T. gamsii YIM PH30019 was nonpathogenic to the host, presented appreciable biocontrol efficacy, and could tolerate chemical fertilizer concentrations of up to 20%. T. gamsii YIM PH30019 displayed antagonistic activities against the pathogenic fungi of P. notoginseng via production of VOCs. On the basis of gas chromatography-mass spectrometry, VOCs were identified as dimethyl disulfide, dibenzofuran, methanethiol, ketones, etc., which are effective ingredients for antagonistic activity. T. gamsii YIM PH30019 was able to improve the seedlings' emergence and protect P. notoginseng plants from soil-borne disease in the continuous cropping field tests. Conclusion: The results suggest that the endophytic fungus T. gamsii YIM PH30019 may have a good potential as a biological control agent against notoginseng phytodiseases and can provide a clue to further illuminate the interactions between Trichoderma and phytopathogens.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.3
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• pp.195-204
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• 1998

This study was designed to investigate the effects of antagonistic microorganism, Bacillus subtilis, on the growth of forage seedlings in repeated cultivation soils and unrepeated cultivation soils. The field experiment was wnducted in pots in a vinyl house using repeated and unrepeated cultivation soils. Forage types were 'Suwon 19' wrn(Zea mqs L.), 'Califbmia' white clover(Tr~oIium repens L.) and 'Fawn' tall fescue (Festuca arundianacea Schreb.). Samples of white clover and tall fescue were taken h m each pot at 36 days after seeding. Samples of wm were examined at 50 days after seeding. The most active antagonistic bacterium was isolated h m forage rhizosphere soil, and selected by reference to it's antagonistic ability on the growth of pathogenic fungi, Rhizoctonia solmi and Fusarium oxyspomm, and it was identified as Bacillus subtilis. This strain strongly suppressed the growth of fungal pathogens among isolated rhizobacteria. The dry weight of forage shoots and roots cultivated in unrepeated cultivation soils was higher than that cultivated in repeated cultivation soils. The dry weight of forage was positively affected by the inoculation of the antagonistic bacterium, Bacillus subtilis, in both repeated cultivation soils and unrepeated cultivation soils. In conclusion, the growth of forage was more affected by the inoculation of the antagonistic bacterium in unrepeated cultivation soils than that in repeated cultivation soils, and bacterization of forage with B. subtilis resulted in an inrreased yield.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.3
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• pp.239-248
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• 1997

This study was conducted to investigate the effects of antagonistic microorganisms, Bacillus spp., on growth of alfalfa(Medicag0 sativa L.) in repeated cultivation soil(RCS) and unrepeated cultivation soil(URCS). Alfalfa was established by seeding into pots 12 cm in diameter and 9 cm in depth containing 1 : 1 mixture of soil and vermiculite with antagonistic bacteria and pathogenic fungi. The growth experiment of alfalfa was conducted in pots in a vinyl house. The bacteria used in this study were Bacillus subtilis and hsants. B. subtilis was isolated and identified 60m forage rhizosphere soil and hsants isolated through cell fusion fiom B. subtilis 101 and B. thuringiensis. B. subtilis was named B. subtilis 101 and hsants named F -3 and F -8. From dark culture experimenf alfalfa was longer lived in treated soil with antagonistic bacteria than that in non-treated soil, and longer lived in URCS than that in RCS. However, alfalfa was shorter lived in RCS and URCS than that in autoclaved RCS. The number of leaves of alfalfa were positively affected by the inoculation of the antagonistic bacteria in both RCS and URCS. Dry weight of shoot and root was increased by the inoculation of the antagonistic bacteria(P< 0.05). However, the growth of alfalfa was decreased by the inoculation of the pathogenic hngi both RCS apd URCS.

• The Plant Pathology Journal
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• v.28 no.2
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• pp.156-163
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• 2012

Stem rots caused by Rhizoctonia solani and Sclerotinia sclerotiorum have been known as devastating diseases in balloon flower plants. Antifungal activities of four fungicides, azoxystrobin, polyoxin B, trifloxystrobin and validamycin A were evaluated in vitro, showing effective suppression with mycelial growth of the fungal isolates on PDA media. Efficacies of the four fungicides were also demonstrated in stem tissues of balloon flower plants against R. solani and S. sclerotiorum. A commercially available Bacillus subtilis strain Y1336 was tested in terms of antagonistic biological control of stem rot disease of balloon flower plants. The bacterial strain revealed its antifungal activities against R. solani and S. sclerotiorum demonstrated by dual culture tests using paper discs and two plant pathogenic fungi on PDA media, as well as by plant inoculation assay, indicating that this antagonistic bacterial strain can be incorporated into disease management program for balloon flower stem rot diseases together with the four chemical fungicides.

• Journal of Applied Biological Chemistry
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• v.42 no.3
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• pp.146-150
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• 1999

As a part of the integrated disease system in greenhouse, an antifungal fungus(AF1) was isolated from greenhouse soil. It exhibited strong inhibitory activites against Pythium ultimum, Phytophtora capsici, Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum based on dual culture on 1/5 strength of potato dextrose agar between antagonistic fungus and several plant pathogens. The antagonistic fungus was identified as Chaetomium cochliodes, based on morphological characteristics; the body of the perithecium bears straight or slightly wavy, unbranched hairs, whilst the apex bears a group of spirally coiled hairs. To investigate antagonistic principles, antifungal compound was extracted and fractionated by different solvent systems. An antifungal compound was isolated as pure crystal from is culture filtrate using organic solvent extraction and column chromatography, followed by preparative thin layer chromatography. The chemical structure of the purified antifungal compound was identified as chaetoglobosin A based on the data obtained form $^1H-NMR$, $^{13}C-NMR$, DEPT 90, 135, $^1H-^1H$ COSY, $^1H-^{13}C$ COSY and EI/MS. $ED_{50}$ values of the chaetoglobosin A against P. ultimum, P. capsici, R. solani, B. cinerea and F. oxysporum were 1.98, 4.01, 4.16, 2.67 and 35.14 ppm, respectively.

• Mycobiology
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• v.46 no.4
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• pp.370-381
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• 2018

Bioactive natural compounds, isolated from fungal endophytes, play a promising role in the search for novel drugs. They are an inspiring source for researchers due to their enormous structural diversity and complexity. During the present study fungal endophytes were isolated from a well-known medicinal shrub, Berberis aristata DC. and were explored for their antagonistic and antioxidant potential. B. aristata, an important medicinal shrub with remarkable pharmacological properties, is native to Northern Himalayan region. A total of 131 endophytic fungal isolates belonging to eighteen species and nine genera were obtained from three hundred and thirty surface sterilized segments of different tissues of B. aristata. The isolated fungi were classified on the basis of morphological and molecular analysis. Diversity and species richness was found to be higher in leaf tissues as compared to root and stem. Antibacterial activity demonstrated that the crude ethyl acetate extract of 80% isolates exhibited significant results against one or more bacterial pathogens. Ethyl acetate extract of Alternaria macrospora was found to have potential antibacterial activity. Significant antioxidant activity was also found in crude ethyl acetate extracts of Alternaria alternata and Aspergillus flavus. Similarly, antagonistic activity of the fungal endophytes revealed that all antagonists possessed inhibition potential against more than one fungal pathogen. This study is an important step towards tapping endophytic fungal diversity for bioactive metabolites which could be a step forward towards development of novel therapeutic agents.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.47-47
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• 2014

Dokdo island is located in the northeastern part of Ulleungdo, known as volcanic island. In total, 53 fungal isolates were isolated from Dokdo island soil sample, using dilution plate technique. The isolates were identified on the basis of morphological characteristics and rDNA ITS sequence analysis. Out of them, 41 isolates were identified at the level of species. The dominant fungal species and genera included Fusarium spp., Mucor sp., Clonostachys spp., and Trichoderma sp. The % sequence identity (the number of matches/the complete alignment length) values via NCBI BLAST searching of EML-IF9, EML-MF30-1 and EML-DDSF4 represented 97.19% (485/499) with Clonostachys cf. rosea (GenBank accession no. KC313107), 98.33% (472/480) with Metarhizium guizhouense (GenBank accession no. HM055445), and 100% (350/350) with Mortierella oligospora (GenBank accession no. JX976032), respectively. Three species of C. rosea, M. guizhouense and M. oligospora represented new records of fungi from Dokdo island, Korea. The antimicrobial activities of the fungal strains varied with tested. Two isolates (EML-MFS30-1 and EML-IF9) showed antifungal activity against several fungi including Fusarium oxysporum and Rhizotonia solani. Clonostachys rosea (EML-IF9) showed strong hydrolytic enzyme activity. Our results showed that the antagonistic fungi including Clonostachys rosea will be used as potential biocontrol agents for control of fungal diseases.

• The Plant Pathology Journal
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• v.38 no.1
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• pp.33-45
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• 2022

To screen antagonistic fungi against plant pathogens, dual culture assay (DCA) and culture filtrate assay (CFA) were performed with unknown soil-born fungi. Among the different fungi isolated and screened from the soil, fungal isolate ANU-301 successfully inhibited growth of different plant pathogenic fungi, Colletotrichum acutatum, Alternaria alternata, and Fusarium oxysporum, in DCA and CFA. Morphological characteristics and rDNA internal transcribed spacer sequence analysis identified ANU-301 as Aspergillus terreus. Inoculation of tomato plants with Fusarium oxysporum f. sp. lycopersici (FOL) induced severe wilting symptom; however, co-inoculation with ANU-301 significantly enhanced resistance of tomato plants against FOL. In addition, culture filtrate (CF) of ANU-301 not only showed bacterial growth inhibition activity against Dickeya chrysanthemi (Dc), but also demonstrated protective effect in potato tuber against soft rot disease. Gas chromatography-tandem mass spectrometry analysis of CF of ANU-301 identified 2,4-bis(1-methyl-1-phenylethyl)-phenol (MPP) as the most abundant compound. MPP inhibited growth of Dc, but not of FOL, in a dose-dependent manner, and protected potato tuber from the soft rot disease induced by Dc. In conclusion, Aspergillus terreus ANU-301 could be used and further tested as a potential biological control agent.

• Research in Plant Disease
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• v.24 no.3
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• pp.213-220
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• 2018

We classified the previously reported antagonistic strains of Sorangium cellulosum into 5 subspecies (A-E). Four strains were antagonistic to Botrytis cinerea (AB group) and two strains were antagonistic to Colletotrichum acutatum (AC group). According to the genetic and sequential analyses with standard genes, xynB1, bglA2, groEL1 for grouping, all strains of AB group were belonged to subspecies C and all strains of AC group were belonged to subspecies D. In addition, high pressure liquid chromatography with the culture filtrates confirmed the genetic results, because AB group had peaks with retention time at 20-22.5 minutes, whereas AC group had no peak. There was positive relationship ($R^2=0.9652$) between the control values of infecting B. cinerea on cherry tomatoes and the main peak areas of chromatograms among the four isolates of AB group. From the subspecies results of AB group, the main peak of KYC 3270 was expected to be epothilone D. However the retention times of the standard of commercial epothilone D and the main peak of KYC 3270 culture filtrate were different as 9.9 and 11.581 min., respectively. Finally, the antagonistic metabolite of AB group was inferred as 7-ketone epothilone D.