• The Plant Pathology Journal
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• v.23 no.3
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• pp.180-186
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• 2007

The biocontrol agent, Serratia plymuthica A21-4, has been developed for controlling pepper Phytophthora blight. Serratia plymuthica A21-4 strongly inhibits the mycelial growth, zoospore formation, and cyst germination of Phytophthora capsici in vitro. The application of a cell suspension of strain A21-4 to pepper plants in pot experiments and in greenhouse successfully controlled the disease. The bacteria produced a potent antifungal substance which was a key factor in the suppression of Phytophthora capsici. The most active chemical com-pound was isolated and purified by antifungal activity-guided fractionation. The chemical structure was identified as a chlorinated macrolide $(C_{23}H_{31}O_8Cl)$ by spectroscopic (UV, IR, MS, and NMR) data, and was named macrocyclic lactone A21-4. The active compound significantly inhibited the formation of zoosporangia and zoospore and germination of cyst of P. capsici at concentrations lower than $0.0625{\mu}g/ml$. The effective concentrations of the macrocyclic lactone A21-4 for $ED_{50}$ of mycelial growth inhibition were $0.25{\mu}g/ml,\;0.25{\mu}g/ml,\;0.30{\mu}g/ml \;and\;0.75{\mu}g/ml$ against P. capsici, Pythium ultimum, Sclerotinia sclerotiorum and Botrytis cinerea, respectively.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1272-1275
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• 2017

Two dimeric sesquiterpenes were separated from Chloranthus japonicus Sieb. and identified as shizukaols C and F. They exhibited potent antifungal activities ($MICs=4-16{\mu}g/ml$) in vitro against various plant pathogenic fungi (Pythium ultimum, Phytophthora infestans, Botrytis cinerea, Colletotrichum lagenarium, Alternaria kikuchiana, and Magnaporthe grisea). Shizukaol C showed 88% and 91% protective activities in the greenhouse against Puccinia recondita (wheat leaf rust) and Phytophthora infestans (tomato late blight), respectively, at $100{\mu}g/ml$; shizukaol F exhibited 93% antifungal activity against Puccinia recondita at the same concentration. Therefore, these compounds might serve as interesting candidates for effective antifungal agents.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.1030-1036
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• 2018

Bacillus strains produce various types of antibiotics, and random mutagenesis has traditionally been used to overproduce these natural metabolites. However, this method leads to the accumulation of unwanted mutations in the genome. Here, we rationally designed a single nucleotide substitution in the degU gene to generate a B. subtilis strain displaying increased plipastatin production in a foreign DNA-free manner. The mutant strain (BS1028u) showed improved antifungal activity against Pythium ultimum. Notably, pps operon deletion in BS1028u resulted in complete loss of antifungal activity, suggesting that the antifungal activity strongly depends on the expression of the pps operon. Quantitative real-time PCR and lacZ assays showed that the point mutation resulted in 2-fold increased pps operon expression, which caused the increase in antifungal activity. Likewise, commercial Bacillus strains can be improved to display higher antifungal activity by rationally designed simple modifications of their genome, rendering them more efficient biocontrol agents.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1994.06a
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• pp.50-61
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• 1994

Biological control of important fungal diseases such as Phytophthora blight of red pepper, gary mold rot of vegetables, and powdery mildew of many crops was attempted using an antagonistic bacterium, Bacillus sp. AC-1 in greenhouses and fields. The antagonistic bacterium isolated from the rhizosphere soils of healthy red pepper plant was very effective in the inhibition of mycelial growth of plant pathogenic fungi in vitro including Phytophthora capsici, Rhizoctonia solani, Pyricularia oryzae, Botrytis cinerea, Valsa mali, Fusarium oxysporum, Pythium ultimum, Alternari mali, Helminthosporium oryzae, and Colletotrichum gloeosporioides. Culture filtrate of antagonistic Bacillus sp. AC-1 applied to pot soils infested with Phytophthora capsici suppressed the disease occurrence better than metalaxyl application did until 37 days after treatment in greenhouse tests. Treatments of the bacterial suspension on red pepper plants also reduced the incidence of Phytophthora blight in greenhouse tests. In farmers' commercial production fields, however, the controlling efficacy of the antagonistic bacteria was variable depending on field locations. Gray mold rot of chinese chives and lettuce caused by Botrytis cinerea was also controlled effectively in field tests by the application of Bacillus sp. AC-1 with control values of 79.7% and 72.8%, respectively. Spraying of the bacterial suspension inhibited development of powdery mildew of many crops such as cucumber, tobacco, melon, and rose effectively in greenhouse and field tests. The control efficacy of the bacterial suspension was almost same as that of Fenarimol used as a chemical standard. Further experiments for developing a commercial product from the antagonistic bacteria and for elucidating antagonistic mechanism against plant pathogenic fungi are in progress.

• Journal of Life Science
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• v.20 no.8
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• pp.1254-1260
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• 2010

Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the $ED_{50}$ value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 mg/ml, 0.16 mg/ml and 0.29 mg/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The $^1H$ and $^{13}C$ assignment of the antagonistic substance was achieved from two-dimensional $^1H-^1H$ COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid ($C_{13}H_8N_2O_2$, M.W.=224).

• Research in Plant Disease
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• v.21 no.1
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• pp.12-19
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• 2015

Antimicrobial activities of nano-materials were tested against several plant pathogens. Twelve different nano-materials were used to observe the antagonistic activity against three kinds of mold and sixteen different kinds of watermelon fruit rot pathogens (Acidovorax citrulli). According to the results, no antagonism have been found against the pathogen, Cylindrocarpon destructans. However in the case of Pythium ultimum, combination of Brass/Glucose 1,000 ppm confirmed the mycelial growth reduction by 94%. In addition, little effect was found against Rhizoctonia solani by Ag/Glucose 3,000 ppm. The remaining other nano-materials have different antimicrobial effect depending on the strains of A. citrulli. But in the case of lime (Cu/Salt 1,000 ppm) highest antimicrobial activity was observed with 97%. Moreover growth of five different strains of A. citrulli was checked by 99% with the combination of Ag/Glucose 1,000 ppm. 92% reduction of A. citrulli growth was observed with $Brass/CaCO_3$ 3,000 ppm. Tested nano-materials against different plant pathogens in this study showed the antimicrobial activity at the range of 24-70%.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1361-1368
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• 2007

Endophytic bacteria associated with the roots of coastal sand dune plants were isolated, taxonomically characterized, and tested for their plant growth-promoting activities. Ninety-one endophytic bacterial isolates were collected and assigned to 17 different genera of 6 major bacterial phyla based on partial 16S rDNA sequence analyses. Gammaproteobacteria represented the majority of the isolates (65.9%), and members of Pseudomonas constituted 49.5% of the total isolates. When testing for antagonism towards plant pathogenic fungi, 25 strains were antagonistic towards Rhizoctonia solani, 57 strains were antagonistic towards Pythium ultimum, 53 strains were antagonistic towards Fusarium oxysporum, and 41 strains were antagonistic towards Botrytis cinerea. Seven strains were shown to produce indole acetic acid (IAA), 33 to produce siderophores, 23 to produce protease, 37 to produce pectinase, and 38 to produce chitinase. The broadest spectra of activities were observed among the Pseudomonas strains, indicating outstanding plant growth-promoting potential. The isolates from C. kobomugi and M. sibirica also exhibited good plant growth-promoting potential. The correlations among individual plant growth-promoting activities were examined using phi coefficients, and the resulting data indicated that the production of protease, pectinase, chitinase, and siderophores was highly related.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.95.2-96
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• 2003

In order to develop a new microbial fungicide for the control of vegetable diseases using endophytic bacteria, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth media, their antifungal activities were screened by in vivo bioassays against Botrytis cinerea(tomato gray mold), Pythium ultimum(cucumber damping-off), Phytopkhora infestans(tomato late blight), Colletotrichum orbiculare(cucumber anthracnose), and Blumeria graminis f. sp. hordei(barley powdery mildew). As the results of screening, 38 bacterial strains showed potent antifungal activities against at least one of 5 plant pathogens. A bacterial strain EB072 displayed potent disease control activities against 3 plant diseases. Among the bacterial strains with a potent antifungal activity against cucunlber anthracnose, three bacterial strains, EB054, EB151 and EB215, also displayed a potent in vitro antifungal activity against C. acutatum, a fungal agent causing pepper anthracnose. A bacterial strain EB215 obtained from roots of cucumber was identified as Burkholderia cepacia based on its physiological and biochemical characteristics and 165 rRNA gene sequence. An antifungal substance was isolated from the liquid cultures of B. cepacia EB215 strain by ethyl acetate partitioning, repeated silica gel column chromatography, and invitro bioassay, Its structural determination is in progress by various instrumental analyses.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.450-456
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• 2006

Pseudomonas fluorescens MC07 is a growth-promoting rhizobacterium that suppresses mycelial growth in fungi such as Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici. To determine the role of the bacterium's antifungal activity in disease suppression, we screened 2,500 colonies generated by Tn5lacZ insertions, and isolated a mutant 157 that had lost antifungal activity. The EcoRI fragment carrying Tn5lacZ was cloned into pBluescript II SK(+) and used as a probe to isolate wild-type clones from a genomic library of the parent strain, MC07. Two overlapping cosmid clones, pEH4 and pEH5, that had hybridized with the mutant clone were isolated. pEH4 conferred antifungal activity to the heterologous host P.fluorescens strain 1855.344, whereas pEH5 did not. Through transposon mutagenesis of pEH4 and complementation analyses, we delineated the 14.7-kb DNA region that is responsible for the biosynthesis of an antifungal compound. DNA sequence analysis of the region identified 11 possible open reading frames (ORF), ORF1 through ORF11. A BLAST search of each putative protein implied that the proteins may be involved in an antifungal activity similar to polyketides.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.984-991
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• 2005

Soil or seed applications of plant growth-promoting rhizobacteria (PGPR) have been used to enhance growth of several crops as well as to suppress the growth of plant pathogens. In this study, we selected a PGPR strain, Paenibacillus polymyxa strain E681, out of 3,197 heat-stable bacterial isolates from winter wheat and barley roots. Strain E681 inhibited growth of a broad spectrum plant pathogenic fungi in vitro, and treatment of cucumber seed with E681 reduced incidence of damping-off disease caused by Pythium ultimum, Rhizoctonia solani, or Fusarium oxysporum. When inoculated onto seeds as vegetative cells or as endospores, E681 colonized whole cucumber root systems and root tips. Different temperatures such as $20^{\circ}C\;and\;30^{\circ}C$ did not affect root colonization by strain E681. This colonization was associated with a consistent increase in foliar growth of cucumber in the greenhouse. These results indicate that strain E681 is a promising PGPR strain for application to agricultural systems, particularly during the winter season.