• Korean Journal of Environmental Biology
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• v.25 no.4
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• pp.356-362
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• 2007

The rhizobacterial diversity associated with 9 native plant species inhabiting coastal sand dunes in Tae-an area, Chungnam Province, was studied using the denaturing gradient gel electrophoresis (DGGE) fingerprinting analysis over three times from October 2003 to March 2004. One dominant band commonly occurred in all of the rhizosphere samples, which was identified as that of Lysobacter enzymogenes. The other common bands included those derived from species of Pseudomonas and Bacillus. It was notable that L. enzymogenes was dominant in all of the 9 plant species and such dominance was consistent throughout the whole sampling period, which confirms the previous study by Lee et al. (2006a). The Bacillus bands were detected in all of the three samplings, and those of Pseudomonas were notable in the samples of December 2003. By the DGGE analysis alone, the significance of Lysobacter to the sand dune plants is not clear. However, considering their presence in healthy plants and the dominance in all plant species, Lysobacter may have positive roles in the survival or growth of the plants in sand dune area.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.490-497
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• 2010

A chitinolytic bacterium having a strong antagonistic activity against various pathogens including Phytophtora capsici was isolated from rhizosphere soil, and identified as Lysobacter enzymogenes (named as LE429) based on 16S rRNA gene sequence analysis. This strain produced a number of substances such as chitinase, ${\beta}-1$, 3-glucanase, lipase, protease, gelatinase and an antibiotic compound. This antibiotic compound was purified by diaion HP-20, sephadex LH-20 column chromatography and HPLC. The purified compound was identified as phenylacetic acid by gas chromatography-electron ionization (GC-EI) and gas chromatography-chemical ionization (GC-CI) mass spectrometry. In field experiment, pepper plants were treated by the strain LE429 culture (CB), neem oil solution (NO), combination (CB+NO) or control (CON). Plant height and number of branches, flowers and pods of pepper plant in CB treatment were generally highest, and followed by CB+NO, CON and NO. The fungal pathogens were strongly inhibited, while several insect pests were discovered in CB treatment. Any insect pests were not found, while all fungal pathogens tested were not suppressed in NO treatment. However, in CB+NO treatment, non incidence of fungal pathogens and insect pests were found. The strain LE429 producing secondary metabolites with neem oil should be a potential agent to control fungal diseases and insect pests.

• Journal of Radiation Industry
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• v.4 no.1
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• pp.65-71
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• 2010

Two chitinase producing strains CHI2 and CHI4 were isolated from soybean rhizosphere soil. Both the strains belonged to Lysobacter enzymogenes as indicated by 16S rDNA sequence analysis. Though strain CHI2 and CHI4 produced extracellular chitinase, they differ in their chitinolytic activity. CHI4 produced approximately three times the higher amounts of enzyme than that of CHI2 under specified conditions. CHI2 produced $535.67U\;l^{-1}$ of chitinase after 48 h incubation with a specific activity of $3.91U\;mg^{-1}$ of protein while strain CHI4 produced $1584.13U\;l^{-1}$ of chitinase with a specific activity of $10.88U\;mg^{-1}$ protein. SDS-PAGE analysis indicated that the molecular weight of chitinase enzyme was approximately 45 kDa. A faint band with a molecular weight of 55 kDa reveals the possibility for the presence of another kind of chitin binding protein. Mutant library was developed by exposing the isolates to gamma rays at their $LD_{99}$ value (0.23 kGy). Totally, 11 mutants of CHI2 and CHI4 are reported to have enhanced chitinase activity. Several leaky mutant clones with decreased enzyme activity and a defective mutant (CHI2-M16) with complete loss of chitinase activity were also identified. CHI4-M18, CHI4-M8 and CHI4-M29 showed 78.8, 41.5, and 31.9% increased chitinase activity over wild type CHI4.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.439-445
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• 2012

The chitinase producing Lysobacter enzymogenes C-3 has previously been shown to suppress plant pathogens in vitro and in the field, but little is known of the regulation of chitinase production, or its role in antimicrobial activity and biocontrol. In this study, we isolated and characterized chitinase-defective mutants by screening the transposon mutants of L. enzymogenes C-3. These mutations disrupted genes involved in diverse functions: glucose-galactose transpoter (gluP), disulfide bond formation protein B (dsbB), Clp protease (clp), and polyamine synthase (speD). The chitinase production of the SpeD mutant was restored by the addition of exogenous spermidine or spermine to the bacterial cultures. The speD and clp mutants lost in vitro antifungal activities against plant fungal pathogens. However, the gluP and dsbB mutants showed similar antifungal activities to that of the wild-type. The growth of the mutants in nutrient rich conditions containing chitin was similar with that of the wild-type. However, growth of the speD and gluP mutants was defective in chitin minimal medium, but was observed no growth retardation in the clp and dsbB mutant on chitin minimal medium. In this study, we identified the four genes might be involved and play different role in the production of extracellular chitinase and antifungal activity in L. enzymogenes C-3.

• Research in Plant Disease
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• v.13 no.1
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• pp.40-44
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• 2007

Powdery mildew of pepper is one of the most devastating diseases which is occurring all the year under greenhouse condition. In this study, control efficacy against powdery mildew was evaluated by mixed culture solutions of two chitinolytic bacteria, Lysobacter enzymogenenes strain C-3 and Chrornobacterium sp. strain C-61, cultivated in the chitin-supplemented medium. In all experiments, white powder on the reverse side of pepper leaves perfectly disappeared 3 days after application of mixed culture solutions. However, periods required for formation of new white powder on the same sites after application (control-lasting period) were largely differed according to environmental conditions. In particular, the control-lasting period was much longer when sprayed on 6 PM than 9 AM and especially, on rainy days than sunny days. This indicates that control efficacy of culture solution may be largely affected by environmental conditions after application. The undiluted culture solution resulted in a perfect control with control value more than 95% by application of 5-day-intervals under severely diseased field and 7-day-intervals under disease-started field. A ten-fold diluted product also showed control value more than 81% by application of the same method. These results suggest that this culture solution can be practically used to control powdery mildew disease in pepper plants.

• Research in Plant Disease
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• v.24 no.4
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• pp.353-358
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• 2018

An effective bioformulation of mixtures of chitin-degrading bacteria has been used successfully to control plant diseases and nematodes. In this study, the bioformulation approach was assessed to control damping-off disease of ginseng. In pot experiments with soils infested with dapming-off pathogens of ginseng, root-drenchings of Chrobacterium sp. C-61, Lysobacterium enzymogenes C-3, and mixture of two bacterial strains grown in chitin minimal medium were signficantly increased emergence of seeds and reduced damping-off disease incidence of seedlings. Efficacy of the bioformulated product depended on the dose and timing of application. In two-year-old ginseng field, the high control efficacies were achieved by soil drenching of two times with an undiluted product or three times with a 10-fold diluted product. In a To-jik nursery (self soil nursery), biocontrol efficacy of the undiluted product against damping-off disease were similar to that of a seed dressing with fungicide, Tolclofos-methyl WP. These results suggest that the bioformulated product containing Chromobacterium sp. C-61 and L. enzymogenes C-3 could be an effective approach to control of ginseng damping-off disease.