• Journal of Microbiology
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• v.39 no.3
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• pp.177-180
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• 2001

Glycinecin A, a bacteriocin produced by Xanthomonas campestris pv. glycines, inhibits the growth of X. c. pv. vesicatoria. We have reported that purified glycinecin A is composed of two polypeptides, is active over a wide range of pH (6 to 9), and is stable at temperatures up to 60$\^{C}$. Glycinecin A is a heterodimer consisting of 39- and 14-kDa subunits; the two encoding genes, glyA and glyB, respectively, have been cloned (Heu et al. 2001. Appl. Environ. Microbiol. 67, 4105-4110). Co-expression of glyA and glyB in the same cell is essential for bacteriocin activity. We constructed and produced a chimeric glycinecin A connecting glyA and glyB in one open reading frame. The chimeric glycinecin A has the same bactericidal activity as the wild-type glycinecin A. However, the chimeric glycinecin A is more stable in a wider range of pH and temperature.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.376-381
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• 1998

Xanthomonas campestris pv. glycines 8ra causes bacterial pustule disease on susceptible soybean leaves and produces a bacteriocin, named glycinecin, against related bacteria such as Xanthomonas campestris pv. vesicatoria. The antimicrobial activity of the glycinecin was effective to most tested Xanthomonas species. X. c. pv. glycines 8ra was able to produce the glycinecin in liquid media as well as solid media. Maximal productivity of glycinecin was obtained at 3$0^{\circ}C$ in the early stationary phase of growth of the X. c. pv. glycines 8ra. The production of glycinecin was not dependent on the initial inoculum level but on cell density. Glycinecin was very sensitive to proteolytic enzymes such as trypsin and proteinase K but resistant to DNase and RNase. The culture supernatant of X. c. pv. glycines 8ra retained some of its antimicrobial activity after 15 min at 6$0^{\circ}C$. It is stable at wide range of pH. The glycinecin showed the bactericidal activity after the adsorption of the glycinecin to the sensitive bacterial cell.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.249-256
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• 2001

Xanthomonas axonopodis pv. glycines 8ra produces a bacteriocin called glycinecinA, which specifically inhibits the growth of bacteria belonging to Xanthomonas species. GlycinecinA was produced by culturing Escherichia coli DH5 containing biosynthetic genes for glycinecinA, and was tested for its control effect against X. vesicatoria on red pepper and X. oryzae pv. oryzae on rice. The bacteriocin activity was much higher in the cell extract than in the supernatant. It reached a maximum level at the stationary phase, ws maintained up to 2 months at room temperature and approximately 10 months at $4^{\circ}$. The optimum concentration of glycinecinA for the control in the greenhouse and in the field was 12,800 AU/ml. In this study, the activity of glycinecinA on rice and red pepper leaves continued for 7-8 days, during which the pathogen populations remained at low levels. Bacterial leaf spot of red pepper and bacterial leaf blight of rice were significantly reduced by the bacteriocin treatments. The control efficacy was as high as, or even higher than, the chemical treatment of copper hydroxide. These results suggest that the bacteriocin is a potential control agent for bacterial diseases.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.57-61
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• 1999

Xanthomonas campestris pv. glycines causes bacterial pustule disease on susceptible soybean leaves and produces a bacteriocin, named glycinecinA, against most xanthomonads including Xanthomonas campestris pv. vesicatoria. One of the 5 isolated DNA regions responsible for bacteriocin production, a 1.7 kb DNA region for the glycinecinA gene, was used as a probe to detect the presence of the homolog DNA in other bacterial strains. Among 55 bacterial strains tested, only X. campestris pv. glycines showed the positive signal with glycinecinA DNA. Two oligomers, heu2 and heu4, derived from a glycinecinA DNA were used to carry out the polymerase chain reaction (PCR) analysis with chromosomal DNA from 55 different bacterial strains including 24 different strains of X. campestris pv. glycines, 9 different pathovars of xanthomonads, and other 22 bacterial strains of different genus and species. By separation of the PCR products on agarose gel, a 0.86 kb DNA fragment was specifically detected when X. campestris pv. glycines was present in the amplification assay. The 0.86 kb fragment was not amplified when DNA from other bacteria was used for the assay. Southern analysis with glycinecinA DNA showed that the PCR signal was obtained with X. campestris pv. glycines isolates from various geographic regions and soybean cultivars. Therefore, the 1.7 kb DNA region for the glycinecinA gene can be used for the pathovar-specific probe for the DNA hybridization and the primers heu2 and heu4 can be used for the pathovar-specific primers for the PCR analysis to detect X. campestris pv. glycines.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.61.1-61
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.17.1-17
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• 1999

• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.169-175
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• 1996

콩 불마름병균 Xanthomonas campestris pv. glycines 8ra는 X. c. pv. vesicatoria에 길항력이 있는 bacteriocin인 glycinecin을 생성 분비한다. Bacteriocin 생성 분비 능력이 있는 콩 불마름병균을 효과적인 생물학적 방제원으로 활용하기 위해서는 좀더 체계적인 연구가 필요하여, bacteriocin 생성에 관계되는 유전자의 분리를 시도하였다. 약 2,000개의 Xanthomonas campestris pv. glycines 8ra cosmid library에서 bacteriocin의 생성 분비 능력을 조사하여 다섯 개의 clone을, pG011, pG0113, pG33과 pG35, 선발하였다. 그중 한 clone pG08을 임의로 선택하여 plasmid DNA를 분리하였다. Plasmid pG08에서 약 6.0 kb의 DNA를 떼어내어 다른 plasmid vector에 넣은 subclone pBL5는 bacteriocin의 생성 분비 능력이 있었다. Plasmid pG08을 제한효소 처리후 다시 접함시켜 만든 몇 개의 subclone과 pBL5의 제한효소 지도를 비교 분석한 결과 약 3.0 kb의 BamHI-HindIII 부분의 DNA가 bacteriocin의 생성에 관계함을 알았다.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.2
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• pp.142-148
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• 2013

Bacterial diseases in soybean are bacterial pustule by Xanthomonas axonopodis pv. glycines, wildfire by Pseudomonas syringae pv. tabaci, bacterial blight by Pseudomonas savastanoi pv. glycines and bacterial brown spot by Pseudomonas syringae pv. syringae in Korea. It is difficult to identify each disease by early symptoms in fields, because the initial symptoms of these diseases are very similar to each other. In this study, we developed multiplex PCR detection method for rapid and accurate diagnosis of bacterial diseases. The glycinecin A of X. axonopodis pv. glycines, the tabtoxin of P. syringae pv. tabaci, the coronatine of P. savastanoi pv. glycines and the syringopeptin of P. syringae pv. syringae have been reported previously. These bacteriocin or phytotoxin producing genes were targeted to design the specific diagnostic primers. The primer pairs for diagnosis of each bacterial diseases were selected without nonspecific reactions. The studies on simultaneous diagnosis method were also conducted with primarily selected 21 primers. As a result, we selected PCR primer sets for multiplex PCR. Sizes of the amplified PCR products using the multiplex PCR primer set consist of 280, 355, 563 and 815 bp, respectively. This multiplex PCR method provides a efficient, sensitive and rapid tool for the diagnosis of the bacterial diseases in soybean.

• Research in Plant Disease
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• v.15 no.2
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• pp.83-87
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• 2009

Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X axonopodis pv. glycines with DNA concentration as low as 200 fg and $1.8{\times}10^3$ cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.