• Research in Plant Disease
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• v.29 no.4
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• pp.405-413
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• 2023

Bacterial blight of carrot caused by Xanthomonas hortorum pv. carotae (Xhc) is one of the serious diseases of carrot, of which control measures has not been still established in the domestic farm. In this study, in order to select effective sterilizer for bacterial blight of carrots, three antibiotics such as streptomycin, oxolinic acid, kasugamycin, two copper compounds like copper hydroxide and copper sulfate basic and three rhizobacteria Burkholderia gladioli MRL408-3, Pseudomonas fluorescens TRH415-2 and Bacillus cereus KRY505-3 were selected to investigate their direct antibacterial effects using artificial media, aiming to identify effective pesticides against Xhc. Among them, treated medium with antibiotics such as streptomycin, oxolinic acid, and the antagonistic rhizobacteria MRL408-3 were formed inhibition zone. The agrochemicals and the rhizobacteria MRL408-3, which showed antibacterial effects on carrot leaves, pre-treated on the carrot leaves and then inoculated with Xhc. High control effects were shown on the carrot leaves pre-treated with both streptomycin and oxolinic acid. Scanning electron microscopy images of the carrot leaf surfaces showed that the population of bacteria decreased significantly on leaves pre-treated with streptomycin and oxolinic acid. From these results, it can be inferred that antibiotics like streptomycin and oxolinic acid exhibit superior control effects compared to other agents. This study provides valuable insights towards establishing an effective control system for bacterial blight of carrot.

• Korean Journal of Soil Science and Fertilizer
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• v.33 no.4
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• pp.292-301
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• 2000

To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.

• Journal of Life Science
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• v.25 no.12
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• pp.1408-1414
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• 2015

In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

• Journal of Applied Biological Chemistry
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• v.55 no.1
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• pp.47-53
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• 2012

Increasing concerns over green farming technology, plant growth promoting rhizobacterium (PGRP) having growth promoting as well as plant disease suppressing properties was recently preferred to use for biological control of plant pathogens infecting plant. We measured the influence of the selected microbial consortium agents-a mixture of PGPR strains-, commercial bio-fungicide, and chemical pesticides on soil microbial community in red pepper field. The activities of soil enzyme such as dehydrogenase, urease, phosphatase, ${\beta}$-glucosidase, and cellulase were analyzed to investigate that of soil microbial community. We also measured plant length, main stem, stem diameter, number of branches and yields of red-pepper in order to observe the red pepper growth promotion. The results of measuring enzyme activities were dehydrogenase 3.5584 ${\mu}g$ TPF $g^{-1}h^{-1}$, urease 15.8689 ${\mu}g$ $NH_4{^-}N$ $g^{-1}h^{-1}$, phosphatase 0.5692 ${\mu}g$ PNP $g^{-1}h^{-1}$, ${\beta}$-glucosidase 2.4785 ${\mu}g$ PNP $g^{-1}h^{-1}$, and cellulase 86.1597 ${\mu}g$ glucose $g^{-1}h^{-1}$ in the soil treated with the microbial consortium agents, so it came out to be very active in the soil. Observing the growth of red-peppers, the main-stem length and the stem diameter were 6.1% and 8.1% higher in the soil treated with the selected microbial consortium agent than the chemical pesticides. After harvesting, yields were 7.3% higher in the soil treated with selected microbial consortium agents than the chemical pesticides. These results showed that microbial consortium agents contribute to increasing soil microbial diversity, growth promoting, and yield of red pepper.

• Applied Biological Chemistry
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• v.50 no.2
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• pp.95-100
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• 2007

The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.447-453
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• 2007

With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

• Korean Journal of Organic Agriculture
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• v.26 no.4
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• pp.677-686
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• 2018

This study was carried out to determine the effect of Kluyvera sp. CL-2 (KACC 91283P) on the growth of watermelon (Citrullus vulgaris L.). The study consisted of three treatments, no treatment (NT), twice application of Kluyvera sp. CL-2 before transplanting (KC1), and five times application around transplanting (KC2). We determined the chemical properties of soil before and after the treatments, and compared the growth characteristics of watermelon among treatments. The treatment of Kluyvera sp. CL-2 at $1.0{\times}10^6cfu\;mL^{-1}$ significantly increased available $P_2O_5$. The organic matter showed to increase for all treatments, while soil pH, exchangeable Ca and Mg tended to decrease for all treatments. The leaf width was increased by 11.6% for KC1 and 26.2% for KC2 compared to NT. But there were no significant differences in yield, leaf length, fruit weight, fruit length, fruit width, and pericarp thickness among treatments. The contents of free sugars such as fructose and glucose were increased by microbial treatments but sucrose was not different from NT. The content of glucose in watermelon was increased by 13.8% in KC1 and 12.8% in KC2 compared to NT. The content of fructose increased by 14.6% in KC1 and by 39.8% in KC2 compared to NT. The results from the study imply that Kluyvera sp. CL-2 can be used to increase sugar content in watermelon.

• Research in Plant Disease
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• v.28 no.2
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• pp.69-81
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• 2022

This study was performed to screen the efficacy of antagonistic bacterial isolates from various sources against the bacterial fruit blotch (BFB) causing pathogen (Acidovorax citrulli) in cucurbit crops. In addition, plant growth promoting traits of these antagonistic bacterial isolates were characterized. Two thousand seven hundred ninety-four microorganisms were isolated from the collected samples. Molecular identification revealed two A. citrulli out of 2,794 isolates. In vitro antagonistic results showed that, among the 28 antagonistic bacterial isolates, 24 and 14 bacterial isolates exhibited antagonism against HPP-3-3B and HPP-9-4B, respectively. Antagonistic and growth promotion characterization of the antagonistic bacterial isolates were further studied. Results suggested that, 4 antagonistic bacteria commonly showed both antagonism and growth promotion phenotypes. Moreover, 3 isolates possessed growth promoting activities. Overall results from this study suggests that BFB causing bacterial pathogen (A. citrulli) was suppressed in in vitro antagonism assay by antagonistic bacterial isolates. Furthermore, these antagonistic bacterial isolates possessed growth promotion and antagonistic enzyme production ability. Therefore, data from this study can provide useful basic data for the in vivo experiments which ultimately helps to develop the eco-friendly agricultural materials to control fruit rot disease in cucurbit crops in near future.

• Journal of Life Science
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• v.34 no.5
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• pp.304-312
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• 2024

This study was conducted to characterize strain Y10, isolated from discarded chicken feathers. Strain Y10 was identified as Bacillus amyloliquefaciens through phenotypic and 16S rRNA gene analysis. B. amyloliquefaciens Y10 exhibited plant growth-promoting activities, including the production of fungal cell-degrading enzymes (cellulase, lipase, protease, and pectinase), siderophores, ammonia, and indoleacetic acid. Furthermore, strain Y10 was able to inhibit the mycelial growth of several phytopathogenic fungi. When 0.1% sucrose as a carbon source and 0.05% casein as a nitrogen source were added to the basal medium, adjusted to pH 10, and cultured at 35℃, the degradation rate of chicken feathers by strain Y10 was about two times higher than that of the basal medium, with the feathers almost completely degraded in four days. Strain Y10 also degraded various keratin substrates, including duck feathers, wool, and human nails. It was confirmed that the feather hydrolyzates prepared using strain Y10 exhibited antioxidant activities, such as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity (EC₅₀ = 0.38 mg/ml) and superoxide dismutase-like activity (EC₅₀ = 183.7 mg/ml). These results suggest that B. amyloliquefaciens Y10 is a potential candidate for the development of bioinoculants and feed additives applicable to the agricultural and livestock industries, as well as the microbiological treatment of keratin waste.