• Applied Biological Chemistry
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• 제46권3호
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• pp.176-182
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• 2003

한국재래간장으로부터 혈전용해효소를 강하게 생산하는 균주를 선발하고 이를 Bacillus subtilis K7로 동정하였다. B. subtilis K7이 생산하는 혈전용해성 protease를 정제하여 분자량을 확인한 결과 21,500 Da이었다. 정제된 효소의 최적반응조건은 $40^{\circ}C$와 pH 9.0이었으며 pH 5.0라서 12.0까지 안정하고 $50^{\circ}C$에서 20분간 열처리한 후에도 50%이상의 효소활성을 가지며 EDTA, CDTA 및 iodoacetat에 실활하는 효소이었다. 이 효소의 fibrin에 대한 Km 값은 $1.8{\times}10^{-2}$ M이었다.

• 한국식물병리학회지
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• 제10권2호
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• pp.112-118
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• 1994

Forth microbial isolates out of 167 isolates from the soil of controlled cultivation areas inhibited mycelial growth of Rhizoctonia solani AG2-1 causing the strawberry bud rot in vitro. Among the isolates, Kr013 and Kr020 showed suppressive effect to R. solani AG2-1 on seedlings of chinese cabbage treated by root immersion, charcoal carrier granule and drenching on 1.0% infested soil in pot. Furthemore, the corresponding effect was also revealed when the charcoal carrier granule of the isolates were treated on the seedling of strawberry that were planted on the planting hole in pot. To examine the effects of biological control in green house, it had been tested the infection rates by using two different treatments. First, the strawberry runner were planted on the nursery soil mixed with 20% charcoal carrier granule of Kr013 and Kr020 isolate respectively, and grown for 20 days before transplanting. Then the young plants form the mother plant were separated and transplanted on the 1.0% infested soil. Another method was that the charcoal carrier of Kr013 and Kr020 isolates applied to planting hole of 1.0% infested soil just before transplanting. Then the young plants were grown for 20 days on the sterilized nursery soil before transplanting. From the results, the effects of biological control was significantly higher on former treatment (e.g. the infection rates were 7.3 and 5.7%, respectively) than on the latter treatment (e.g. the corresponding value were 16.7 and 15.7%, respectively). The antagonistic isolates of Kr013 and Kr020 were respectively identified as Pseudomonas cepacia with the similarity of 55.0% and 60.0% by using the Biolog GN Microplate system.

• The Plant Pathology Journal
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• 제30권3호
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• pp.288-298
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• 2014

We examined the efficacy of a bacterium for biocontrol of the root-knot nematode (RKN) Meloidogyne hapla in carrot (Daucus carota subsp. sativus) and tomato (Solanum lycopersicum). Among 542 bacterial isolates from various soils and plants, the highest nematode mortality was observed for treatments with isolate C1-7, which was identified as Bacillus cereus based on cultural and morphological characteristics, the Biolog program, and 16S rRNA sequencing analyses. The population density and the nematicidal activity of B. cereus C1-7 remained high until the end of culture in brain heart infusion broth, suggesting that it may have sustainable biocontrol potential. In pot experiments, the biocontrol efficacy of B. cereus C1-7 was high, showing complete inhibition of root gall or egg mass formation by RKN in carrot and tomato plants, and subsequently reducing RKN damage and suppressing nematode population growth, respectively. Light microscopy of RKN-infected carrot root tissues treated with C1-7 showed reduced formation of gall cells and fully developed giant cells, while extensive gall cells and fully mature giant cells with prominent cell wall ingrowths formed in the untreated control plants infected with RKNs. These histopathological characteristics may be the result of residual or systemic biocontrol activity of the bacterium, which may coincide with the biocontrol efficacies of nematodes in pots. These results suggest that B. cereus C1-7 can be used as a biocontrol agent for M. hapla.

• The Plant Pathology Journal
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• 제29권1호
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• pp.42-51
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• 2013

One of the most important limiting factors for the production of the grafted cactus in Korea is the qualitative and quantitative yield loss derived from stem rots especially caused by Bipolaris cactivora. This study is aimed to develop microbial control agents useful for the control of the bipolaris stem rot. Two bacteria (GA1-23 and GA4-4) selected out of 943 microbial isolates because of their strong antibiotic activity against B. cactivora were identified as Bacillus subtilis and B. amyloliquefaciens, respectively, by the cultural characteristics, Biolog program and 16S rRNA sequencing analyses. Both bacterial isolates significantly inhibited the conidial germination and mycelial growth of the pathogen with no significant difference between the two, of which the inhibitory efficacies varied depending on the cultural conditions such as temperature, nutritional compositions and concentrations. Light and electron microscopy of the pathogen treated with the bacterial isolates showed the inhibition of spore germination with initial malformation of germ tubes and later formation of circle-like vesicles with no hyphal growth and hyphal disruption sometimes accompanied by hyphal swellings and shrinkages adjacent to the bacteria, suggesting their antibiotic mode of antagonistic activity. Control efficacy of B. subtilis GA1-23 and B. amyloliquefaciens GA4-4 on the cactus stem rot were not as high as but comparable to that of fungicide difenoconazole when they were treated simultaneously at the time of pathogen inoculation. All of these results suggest the two bacterial isolates have a good potential to be developed as biocontrol agents for the bipolaris stem rot of the grafted cactus.

• 생명과학회지
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• 제13권6호
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• pp.843-848
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• 2003

가금티프스는 가금류에 Salmonella gallinarum이 원인균이 되어 발병하는 양계산업에 막대한 지장을 주는 질병이다. 가금티프스를 억제하기 위한 생균제 개발을 위한 목적으로써 가금티프스 원인균인 Salmonella gallinarum의 생육을 저해시킬 수 있는 길항균주를 토종닭의 내장으로부터 분리하여 생육특성과 길항물질 생산성을 조사하고, 이 균주를 분류학적으로 동정하였다. 분리된 길항균주는 Bacillus amyloliquefaciens와 98% 상동성을 나타내어 최종적으로 Bacillus amyloliquefaciens Y3로 명명하였다. 0.3% maltose, 0.2% $NH_4Cl,\; 37^{\circ}C$ pH 7에서 균생육 및 길항물질의 생산능이 가장 우수하였으나 장내 담즙에 대한 내성은 크게 나쁘지 않을 것으로 확인되어졌다. 생산된 길항물질을 추정하여 본 결과 분자량이 10,000보다 작은 저분자물질이었으며 $80^{\circ}C$에서 20분간 열처리한 후에도 80%의 활성을 유지하는 내열성 물질임을 확인할 수 있었다. 향후 선발되어진 Bacillus amyloliquefaciens Y3가 생산해내는 길항물질에 대한 연구와 개량을 통하고 장내 정착성 실험을 거쳐 우수균주로 확인되면 양계산업에 사용될 우수한 생균제로 개발이 가능할 것이라 생각된다.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.643-650
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• 2000

A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

• 한국유기농업학회지
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• 제19권spc호
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• pp.190-193
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• 2011

Organic and chemical fertilizer amendments are an important agricultural practice for increasing crop yields. In order to maintain the soil sustainability, it is important to monitor the effects of fertilizer applications on the shift of soil microorganism, which control the cycling of many nutrients in the soils. Here, culture-dependent and culture-independent approaches were used to analyze the soil microorganism and community structure under six fertilization treatments, including green manure, rice straw compost, rapeseed cake, pig mature compost, NPK +pig mature compost, NPK and control. Both organic and chemical fertilizers caused a shift of the cultural microorganism CFUs after treatments. Bacterial CFUs of the organic fertilization treatments were significantly higher than that of chemical fertilization treatments. The DGGE profiles of the bacterial communities of the samples showed that the green manure treatment was a distinct difference in bacterial community, with a greater complexity of the band pattern than other treatments. Cluster analyses based on the DGGE profile showed that rice straw compost and pig mature compost had a similar banding pattern and clustered together firstly. Rapeseed cake, NPK, NPK +pig manure compost and control clustered together in other sub-cluster and clearly distinguished from green manure.

• 한국식품위생안전성학회:학술대회논문집
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• 한국식품위생안전성학회 1996년도 제11회 학술대회 및 정기총회 - 식품의 위생 안전성에 관한 최근 연구 동향
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• pp.33-33
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• 1996

Interests in the field of rapid methods and automation in microbiology have been growing steadily on an international scale in recent years. International meetings concerned this problem have been held in elsewhere in the world countries since the past twenty years. But, unfortunately in the field of microbial examination in food hygiene, this problem have not yet been developed so much as in the field of clinical microbiology. Today, I would like to introduce you here present aspects of rapid and automation technologies, those which are manly carrying in milk and meats industries. My illustration will be given recent improved technologies using automatic apparatus and instruments along with process of microbial count procedure. Recent direct microbiological counting system (ChemeScan \ulcorner) as real time ultrasensitive analysis created by Cheminex Ltd., France is now most evolutional instrument to provide direct microbial counts, down to one cell, within 30 minutes. The results from these evaluations how a good correlation between the ChemScan system and the standard plate count method. This system will be successful application for not only in the field of pharmacology but also food microbiology. In addition, current identification of microbes by sophisticated instruments suitable for food microbiology, one of which Biology is manual system (BIOLOG\ulcorner), provides reference-level capability at a modes price. For the manual system, the color reactions in the microplate are read by eye and manually keyed into personal computer. Species identification appears on the computer screen within seconds, along with biotype patterns, a list of closely related species, and other useful statistics. In present this is useful application for microbial ecology and epidemiological survey. RiboPrinter system newly produced by DuPont is now focusing among microbiologists in the world, and is one of the biggest microbial characterization system using a DNA-based approach. The technology analyzer is bacterial culture for its genetic fingerprint or riboprint pattern. Finally Bio-cellTracer system for automatic measurement of fungal growth and Fukitori-Maseter, a Surface Hygiene Monitoring Kit by using swabe procedure in food processing environment are briefly illustrated in this presentation.

• The Plant Pathology Journal
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• 제37권2호
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• pp.137-151
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• 2021

The present study describes the bacterial blight of walnut, caused by Xanthomonas arboricola pv. juglandis (Xaj) in the northern Gyeongbuk province, Korea. Disease symptoms that appear very similar to anthracnose symptoms were observed in walnut trees in June 2016. Pathogens were isolated from disease infected leaves, fruits, shoots, bud, flower bud of walnut, and cultured onto yeast dextrose carbonate agar plates. Isolated bacteria with bacterial blight symptoms were characterized for their nutrient utilization profiles using Biolog GN2 and Vitek 2. In addition, isolates were subjected to physiological, biochemical, and morphological characterizations. Furthermore, isolates were identified using 16S rDNA sequence analysis, and multi-locus sequence analysis using atpD, dnaK, efp, and rpoD. To confirm pathogenicity, leaves, fruits, and stems of 3-year-old walnut plants were inoculated with bacterial pathogen suspensions as a foliar spray. One week after inoculation, the gray spots on leaves and yellow halos around the spots were developed. Fruits and stems showed browning symptoms. The pathogen Xaj was re-isolated from all symptomatic tissues to fulfill Koch's postulates, while symptoms were not appeared on control plants. On the other hand, the symptoms were very similar to the symptoms of anthracnose caused by Colletotrichum gloeosporioides. When walnut plants were inoculated with combined pathogens of Xaj and C. gloeosporioides, disease symptoms were greater in comparison with when inoculated alone. Xaj population size was more in the month of April than March due to their dormancy in March, and sensitive to antibiotics such as oxytetracycline and streptomycin, while resistant to copper sulfate.

• The Plant Pathology Journal
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• 제39권1호
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.