• Research in Plant Disease
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• v.12 no.3
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• pp.267-271
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• 2006

Botrytis cinerea Pers. was found to be highly virulent to the grapevine plant, especially in greenhouse condition. Pseudomonas species play key roles for the biocontrol of many plant diseases especially in soil. Of the 83 isolates of Pseudomonas spp., a bacterial strain P84, isolated from tomato rhizosphere, was shown to suppress a wide range of phytopathogenic fungi in vitro. The isolate was identified as Pseudomonas putida on the basis of its bacteriological and genetic characteristics. The P. putida P84 strain carry the phlD gene for 2,4-diacetylphloroglucinol biosynthesis and may produce the antibiotics as an antagonistic mechanism involved in biocontrol. The antagonistic activity of the bacterium has a promising implication for its use as a biocontrol agent to control grapevine gray mold.

• The Plant Pathology Journal
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• v.28 no.1
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• pp.68-74
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• 2012

Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

• Journal of Korean Society on Water Environment
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• v.32 no.6
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• pp.600-627
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• 2016

Korea is one of the countries with a large veterinary antibiotics market, although antimicrobial resistance in bacteria is becoming a serious issue in many countries. The Korean government started to take interest in estimating the effects of livestock manure on rivers and agricultural soils and in monitoring of heavy metals, organic pollutants and antibiotics in the ambient water and soil. In this paper, pre-treatment methods to separate the selected antibiotics from solid samples were reviewed. It is essential to select an efficient and appropriate procedure for pre-treatment due to the high proportion of proteins and organics in biosolid samples. Pre-treatment consists of extraction followed by clean-up. Initially, homogenized samples were extracted by sonication, mechanical agitation or pressurized liquid extraction with methanol/acetonitrile/water mixture under acidic/basic conditions depending on the compound. However, aminoglycosides and colistin were extracted with 5% trichloroacetic acid and HCl, respectively. Since the ${\beta}-lactams$ are easily decomposed in acidic and basic conditions, they were extracted in neutral pH. Filtration with a membrane (pore size, $0.2{\mu}m$) or solid phase extraction with HLB and methanol, as eluents, was normally applied for the clean-up. At least, three different pre-treatment procedures should be adopted to screen all the selected antibiotics in solid samples.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.92-100
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• 1996

For the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV)) from butyric acid and valeric acid, 10 strains of bacteria capable of producing P(3HB-co-3HV) were isolated from soil. Among them, the strain HJ-067 showed the best ability of producing P(3HB-co-3HV), and was indentified as a Azotobacter sp. For the production of P(3HB-co-3HV), the optimum concentrations of butyric and valeric acid were 3.0g/l, respectively. The most effective nitrogen source was $(NH_4)_{2}SO_4$ at an optimum concentration of 0.75g/l, which was equivalent to 21.36 in C/N ratio. Deficiency of the cationic metal ions ($Zn^{2+},\;Co^{2+},\;Mn^{2+}$) in the proguction medium had stimulating effect on P(3HB-co-3HV) accumulation, especially in the manganese. deficient medium. The optimum temperature for P(3HB-co-3HV) production was 27$^{\circ}C$ and the optimum initial pH was 7.0. Under the optimum conditions, 1.82g/l of P(3HB-co-3HV) and 3.00g/l of dry biomass were produced after 36 hour cultivation, and the P(3HB-co-3HV) yield and HV% were 60.60% (w/w), 15.92%, respectively.

• Microbiology and Biotechnology Letters
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• v.14 no.3
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• pp.239-244
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• 1986

To develop the potential use as new host strain for gene cloning, alkaline-tolerant isolates from soil were examined for amylase activity, protease activity, antimicrobial activity and transformability by using plasmid pUB 110. Of these strains, one was selected and identified as Bacillus sp. YA-14. in the enzymatic properties of Bacillus sp. YA-14 the optimal conditions for the reaction of amylase and protease were at pH 0.8 and pH 7.5 respectively. The antimicrobial activity of Bacillus sp. YA-14 was also found. For the transformation, Bacillus sp. YA-14 was cultured to late logarithmic growth phase ai 37$^{\circ}C$ in modified SPI medium (pH 8.0) containing 0.4% MgSO$_4$. The presence of pUB 110 plasmid DNA in transformants was confirmed by electrophoresis and stably maintained in the new host.

• Applied Biological Chemistry
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• v.34 no.2
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• pp.180-186
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• 1991

A water soluble antifungal antibiotic, Solumycin, was separated from the culture broth of Streptomyces sp. LAM-593, isolated from soil, by butanol extraction, alumina-, 1st and 2nd Sephadex LH-20 column chromatography. The substance was pale yellow crystal which gave a single spot at Rf value 0.24 with ethanol-ammonia water-water (8:1:1), 0.46 with butanol-ethanol-water (5:1:4), 0.84 with 50% methanol on silica gel TLC. It was dissolved well in water, methanol and acidic aq. butanol but not in ethanol, acetone, ethyl acetate, chloroform. acetic acid etc., and gave positive Fehling and Molish reaction. The UV spectrum in methanol showed absorption at 342, 361, 380, and 404 nm. The antibiotic was active against fungi such as Candide, Cryptococcus, Saccharomyces, Trichophyton and Trichosporon, but not to bacteria such as Bacillus, Escherichia and Staphylococcus.

• The Plant Pathology Journal
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• v.23 no.4
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• pp.239-244
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• 2007

MicroTom is a miniature tomato plants with various properties that make it as a model system for experiments in plant molecular biology. To extend its utility as a model plant to study a plant - bacterial wilt system, we investigated the potential of the MicroTom as a host plant of bacterial wilt caused by Ralstonia solanacearum. We compared the disease progress on standard tomato and MicroTom by two inoculation methods, root dipping and soil drenching, using a race 1 strain GMI1000. Both methods caused the severe wilting on MicroTom comparable to commercial tomato plant, although initial disease development was faster in root dipping. From the diseased MicroTom plants, the same bacteria were successfully reisolated using semiselective media to fulfill Koch's postulates. Race specific and isolate specific virulence were investigated by root dipping with 10 isolates of R. solanacearum isolated from tomato and potato plants. All of the tested isolates caused the typical wilt symptom on MicroTom. Disease severities by isolates of race 3 was below 50 % until 15 days after inoculation, while those by isolates of race 1 reached over 50% to death until 15 days. This result suggested that MicroTom can be a model host plant to study R. solanacearum - plant interaction.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.350-355
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• 2013

Plants protect themselves from diverse potential pathogens by induction of the immune systems such as systemic acquired resistance (SAR). Most bacterial plant pathogens thrive in the intercellular space (apoplast) of plant tissues and cause symptoms. The apoplastic leaf exudate (LE) is believed to contain nutrients to provide food resource for phytopathogenic bacteria to survive and to bring harmful phytocompounds to protect plants against bacterial pathogens. In this study, we employed the pepper-Xanthomonas axonopodis system to assess whether apoplastic fluid from LE in pepper affects the fitness of X. axonopodis during the induction of SAR. The LE was extracted from pepper leaves 7 days after soil drench-application of a chemical trigger, benzothiadiazole (BTH). Elicitation of plant immunity was confirmed by significant up-regulation of four genes, CaPR1, CaPR4, CaPR9, and CaCHI2, by BTH treatment. Bacterial fitness was evaluated by measuring growth rate during cultivation with LE from BTH- or water-treated leaves. LE from BTH-treatment significantly inhibited bacterial growth when compared to that from the water-treated control. The antibacterial activity of LE from BTH-treated samples was not affected by heating at $100^{\circ}C$ for 30 min. Although the antibacterial molecules were not precisely identified, the data suggest that small (less than 5 kDa), heat-stable compound(s) that are present in BTH-induced LE directly attenuate bacterial growth during the elicitation of plant immunity.

• Korean Journal of Veterinary Service
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• v.38 no.4
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• pp.211-220
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• 2015

Many chemical disinfectants are using to protect the foot and mouth disease (FMD) and avian influenza (AI) in Korea since 2000. This study was performed to confirm disinfective ability of commercialized chemical disinfectants and to investigate the sterilizing ability of E-ball as alterative to chemical disinfectants. 4 kinds of acidulant, 3 kinds of aldehyde, 1 kind of oxidizer and 300 g of E-ball were used in this study. Dilution rate of disinfective power of all chemical disinfectants were to 200 times. The sterilizing ability of aldehydes were better than the acidulant and oxidizer with Salmonella typhimurium. The sterilizing ability of E-ball treated solution was guessed due to the friction of E-ball deads. In the case of the friction of 2 beads of E-ball, Salmonella typhimurium was sterilizted on $1{\times}10^6/mL$ CFU in the E-ball treated solution. The E-ball treated solution had superior sterizing power compared with the chemical disinfectants in the bacteria of soil for antibacterial examination. E-ball treated solution has a possibility as the substitute of chemical disinfectants to protective the animal diseases contains FMD, AI.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.45-49
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• 2002

Bradyrhizobium japonicum is a soil bacterium with a unique ability to infect the roots of leguminous plants and establish a nitrogen-fixing symbiosis, which has been used as a microbial manure. In this study, we examined the stress response after pretreatment of cells with cold temperature. When pre-treated with cold temperature ($4^{\circ}C$) for 16 hr, B. japonicum increased the viability in subsequent stress-conditions such as alcohol, $H_2O_2$, heat, and dehydration. For cold adpatation, cultured B. japonicum was exposed to $4^{\circ}C$. Upon subsequent exposure to various conditions, the number of adapted cells pretreated by cold adaptation was 10-1000 fold higher than that of non-adaptated ones. It appeared de novo protein synthesis occurred during adaptation, because a protein synthesis inhibitor, chloramphenicol abolished the increased stress tolerance. By using a degenerate PCR primer set, a csp homolog was amplified from B. japonicum genome and sequenced. The deduced partial amino acid sequence of the putative Csp (Cold shock protein) shares a significant similarity with known Csp proteins of other bacteria.