• Journal of Microbiology
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• 제39권3호
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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.

• 한국식물병리학회지
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• 제14권5호
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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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• 제17권5호
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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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• 제15권1호
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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.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1999년도 임시총회 및 추계 학술발표회
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• pp.61.1-61
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• 1999

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1999년도 임시총회 및 추계 학술발표회
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• pp.17.1-17
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• 1999

• 한국작물학회지
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• 제58권2호
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• pp.142-148
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• 2013

국내 콩에서 발생하는 세균병해인 불마름병, 들불병, 세균점무늬병, 세균갈색점무늬병의 다중 진단을 위한 PCR 방법을 요약하면 다음과 같다. 1. 콩에 발생하는 각각의 세균들은 서로 다른 박테리오신(bacteriocin) 이나 파이토톡신(phytotoxin)을 생산하는데 이와 관련한 유전자를 목적으로 하여 진단프라이머를 설계하였다. 2. 불마름병은 glycinecin A, 들불병은 tabtoxin, 세균점무늬병은 coronatine과 세균갈색점무늬병은 syringopeptin을 목적유전자로 하여 다중 진단프라이머 조합을 설계하였다. 3. 1차 선발로 각각의 균주에 대한 단일 진단 프라이머를 선발하였으며, 여기선 선발된 21개의 프라이머들을 조합하여 4종 다중진단프라이머 선발을 위한 2차 선발에 이용하였다. 최종적으로 280 bp의 불마름병, 355 bp의 세균갈색점무늬병, 563 bp의 들불병과 815 bp의 세균점무늬병으로 구성된 다중진단 프라이머 조합이 개발되었다. 4. 선발된 4종 다중 진단 프라이머 조합의 경우 다른 세균들과의 비특이적 반응이 있는지 확인하기 위한 3차 선발을 거쳐 그 특이성을 검증하였다.

• 식물병연구
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• 제15권2호
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• pp.83-87
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• 2009

콩 불마름병을 일으키는 Xanthomonas axonopodis pv. glycines를 종자에서 DNA 추출없이 바로 검출하는 방법에 대하여 연구하였다. 콩 종자에서 X. axonopodis pv. glycines를 특이적으로 검출하기 위해 특이적인 유전자로 알려져 있는 glycinecin A로부터 증폭 size가 401 bp인 primer Xag F1 & Xag R1을 고안하였다. Xag Fl과 Xag R1 primer는 콩 종자와 잎에서 분리한 균주와 KACC에서 분양받은 X. axonopoids pv. glycines 균주를 증폭시켰으나 근연종인 X. axonopodis pv. citri, X. axonopodis pv. vesicatoria 등은 증폭되지 않았다. 콩 종자에 존재하는 것으로 알려진 다른 세균들 역시 증폭되지 않았다. 고안된 Xag F1 & Xag R1 primer를 이용한 X. axonopodis pv. glycines의 검출 한계 농도 측정은 genomic DNA와 세포 현탁액을 이용하였다. X. axonopodis pv. glycines의 genomic DNA는 200 fg까지 증폭이 되었으며, 세포현탁액은 $OD_{600nm}$ 0.1로 농도를 조정한 뒤, $10^{-8}$까지 희석하여 측정한 결과 $10^{-6}$인 $1.8{\times}10^3$ cfu/ml까지 증폭이 확인되었다. 자연 감염된 종자에서 병원균을 검출하기 위해 종자를 육안상 건전종자와 불건전한 종자(변색립, 피해립)로 구분하여 direct PCR 방법으로 실험 한 결과 육안상 건전종자에서는 병원균이 검출되지 않았으나 불건전한 종자에서는 병원균의 검출이 확인되었다. 또한 진탕 배양 시간에 따른 병원균의 검출여부를 조사한 결과 진탕 배양 2시간부터 병원균의 검출이 확인되었으며 시간이 지날수록 증폭 밴드가 더욱 선명해 지는 것을 확인할 수 있었다. 그러므로 고안된 Xag Fl & Xag R1 primer를 이용한 direct PCR 방법은 다른 많은 미생물들로 오염되어진 콩종자에 있는 X. axonopodis pv. glycines를 신속하고 민감하게 검정할 수 있는 효과적인 방법으로 활용될 수 있을 것이다.