• 식물병연구
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• 제14권1호
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• pp.10-14
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• 2008

Ralstonia solanacearum에 의한 풋마름병은 세계적으로 치명적인 병해중의 하나이며 우리나라에서는 가지, 토마토, 감자, 고추 등 가지과 작물에 발병이 보고되어 있다. 2005년${\sim}$2006년 2년 동안 가지 주재배단지의 풋마름병 발생 포장에서 48개의 균주를 수집하여 분리 배양하였으며 R. solanacearum의 특이적인 flipcF/flipcR primer를 이용하여 PCR을 실시한 결과 470-bp의 풋마름병 특이적 밴드가 관찰되어 R. solanacearum으로 동정하였다. 분리 동정된 15균주로 각 기주별 병원성을 검정하였을 때 가지, 토마토에 병원성을 나타내었으나 고추는 저항성을 나타내었다. 가지 풋마름병원균의 biovar분화는 biovar 3, 4로 동정되었으며 특히 biovar 3는 가지 풋마름병 저항성 대목에 다소 강한 병원성을 나타내어 저항성대목의 육종이 필요한 것으로 판단된다.

• The Plant Pathology Journal
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• 제32권1호
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• pp.58-64
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• 2016

Bacterial wilt of tomatoes caused by Ralstonia solanacearum is a devastating disease that limits the production of tomato in Korea. The best way to control this disease is using genetically resistant tomato plant. The resistance degree to R. solanacearum was evaluated for 285 tomato accessions conserved in the National Agrobiodiversity Center of Rural Development Administration. These accessions of tomato were originated from 23 countries. Disease severity of tomato accessions was investigated from 7 days to 14 days at an interval of 7 days after inoculation of R. solanacearum under greenhouse conditions. A total of 279 accessions of tomato germplasm were susceptible to R. solanacearum, resulting in wilt and death in 70 to 90% of these plants. Two tomato accessions were moderately resistant to R. solanacearum. Only four accessions showed high resistance against R. solanacearum. No distinct symptom of bacterial wilt appeared on the resistant tomato germplasms for up to 14 days after inoculation of R. solanacearum. Microscopy of resistant tomato stems infected with R. solanacearum revealed limited bacterial spread with thickening of pit membrane and gum production. Therefore, these four resistant tomato germplasms could be used in tomato breeding program against bacterial wilt.

• 한국환경과학회지
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• 제23권6호
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• pp.1165-1173
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• 2014

For the field application of dielectric barrier discharge plasma reactor in nutrient solution culture, a filtration-DBD (dielectric barrier discharge) plasma reactor was investigated for the Ralstonia solanacearum which causes bacterial wilt in aquiculture. The filtration-DBD plasma reactor system of this study was consisted of filter, plasma reactor, reservoir. The DBD plasma reactor consisted of a quartz dielectric tube, discharge electrode (inner) and ground electrode (outer). The experimental results showed that the inactivation of R. solanacearum with filter media type in filter reactor ranked in the following order: anthracite > fiber ball > sand > ceramic ball > quartz ceramic. In filtration + plasma process, disinfection effect with the voltage was found to small. In disinfection time of 120 minutes, residual R. solanacearum concentration was 1.17 log (15 CFU/mL). When the continuous disinfection time was 120 minute, disinfection effect was thought to keep the four days. In sporadic operation mode of 30 minutes disinfection - 24 hours break, residual R. solanacearum concentration after five days was 0.3 log (2 CFU/mL). It is considered that most of R. solanacearum has been inactivated substantially.

• 미생물학회지
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• 제43권3호
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• pp.179-185
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• 2007

본 연구는 polymerase chanin reaction(PCR)기법과 DNA enzyme-linked immunosorbent assay(DNA ELISA) 기법을 이용하여 토양내 식물병원균인 Ralstonia solanacearum를 검출하고자 하였다. 토양 시료로부터 분석에 사용될 R. solanacearum DNA를 추출하기 위하여 몇 가지 방법을 비교 평가한 결과 기존의 DNA 추출 방법에 비하여 Guanidin isothiocyanate와 Chelex-100 resin을 사용하는 방법 이 토양 내에 존재하는 다양한 중류의 반응 저해 물질과 R. solanacearum만의 고유한 PCR반응 저해물질들을 제거하는 데에 효과적이었다. R. solanacearum만을 특이적으로 검출하기 위해 fliC유전자 부위에 특이적인 몇 종의 primer들을 제작하였다. 이들 중 높은 민감도와 특이도를 나타내는 두 set의 primer RsolfliC(forward; 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J. $Sch\ddot{o}nfeld$ et al.)와 RS_247 (forward; 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3, designed by this study)를 선정하여 nested PCR을 수행할 수 있도록 고안하였다. Nested PCR primer에 biotin을 표지하였고 nested PCR산물의 내부 서열과 특이적으로 교잡반응을 할 수 있는 probe를 제작하여 PCR 결과를 DNA-EIA반응으로 확인 분석할 수 있도록 하였다. Primary PCR과 nested PCR의 산물을 전기영동 상에서 확인한 결과, nested PCR이 약 $10^2$정도의 높은 민감도를 나타내었고 DNA-EIA의 경우 $10^2P{\sim}10^3$정도의 민감도를 상승시켜주는 것으로 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1765-1771
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• 2007

A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.

• 식물병연구
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• 제17권2호
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• pp.184-190
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• 2011

Ralstonia solanacearum은 풋마름병(bacterial wilt)을 일으키는 종자 전염 병원세균으로 한번 발생하면 방제가 매우 어려운 병이다. 따라서 Ralstonia solanacearum는 한국을 비롯한 여러 나라에서 식물검역병으로 지정되어있다. 본 연구에서는 가지과 종자로부터 Ralstonia solanacearum 검출할 수 있는 PCR 방법을 개발하였다. 프라이머 RSJH-F와 RS-JH-R는 오직 Ralstonia solanacearum race 1, 3으로부터 401 bp 크기의 DNA를 증폭하였다. 1차 PCR 증폭산물의 내부에서 디자인 된 nested PCR 프라이머인 RS-JH-F-ne and RS-JH-R-ne는 오직 Ralstonia solanacearum로부터 131 bp 크기의 DNA를 증폭하였다. 이들 프라이머들은 토마토, 고추를 포함한 가지과 종자 추출액으로부터 어떤 비특이적 DNA도 증폭하지 않았다. 인공적으로 병원균을 접종한 가지과 종자를 이용하여 병원균 검출 민감도를 비교하였을 때, 본 연구에서 개발한 nested PCR 방법이 ELISA나 반선택배지보다 100배 높은 민감도를 보여주었다. 따라서, 본 연구에서 개발한 PCR 방법들은 가지과 종자로부터 Ralstonia solanacearum를 검출하는 매우 유용한 방법으로 생각된다.

• 한국환경과학회지
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• 제21권7호
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• pp.797-804
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• 2012

A dielectric barrier discharge (DBD) plasma reactor was investigated for the inactivation of Ralstonia Solanacearum which causes bacterial wilt in aquiculture. The DBD plasma reactor of this study was divided into power supply unit, gas supply unit and plasma reactor. The plasma reactor consisted of a quartz dielectric tube, discharge electrode (inner) and ground electrode (outer). The experimental results showed that the optimum 1st voltage, 2nd voltage, air flow rate and pH were for 100 V (1st voltage), 15 kV (2nd voltage), 4 L/min, and pH 3, respectively. At a low 1st voltage, shoulder and tailing off phenomena was observed. The shoulder phenomenon was decreased as the increase of 1st voltage. R. Solanacearum disinfection in the lower air flow rate was showed shoulder and tailing off phenomenon because the active species generated less. Under optimum condition, shoulder and tailing off phenomenon was reduced. When the 2nd voltage was less than 7.5 kV, tailing off phenomenon was observed and this was not vanishes even though the increase of the disinfection time. The inactivation efficiency increased as the increase of air flow rate, however, the efficiency decreased when the air flow rate was above 4 L/min. R. Solanacearum disinfection at pH 3 showed somewhat higher than in pH 11. The pH effect of R. Solanacearum deactivation is less than the impact on other factor.

• 한국식물병리학회지
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• 제3권3호
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• pp.203-209
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• 1987

Pseudomonas solanacearum의 병원성균주 및 비병원성균주를 담배의 뿌리에 접종하여 그 잎과 줄기, 뿌리에서 P. solanacearum과 Erwinia carotovora subsp. carotovora, Escherichia coli에 대한 항균물질을 분리하였다. 담배즙액을 TLC plate에 발전시킨 결과 비병원성균주에 접종된 담배에서는 $R_f$ 0.6 부근에서, 병운성균주에 접종된 담배에서는 $R_f$ 0.9 부근에서 각각 항균물질을 분리할 수 있았다. 그러나 고압살균된 담배즙액배지에 병원성균주 및 비병원성균주를 3일간 배양한 배양여액에서는 항균물질의 생성이 인정되지 않았다. 비병원성균주 및 병원성균주에 접종된 담배잎에서는 무처리 담배에 비하여 peroxidase와 polyph-enoloxidase의 활성이 높았으나, 그 줄기와 뿌리에서는 효소활성의 차이가 없었다. 비병원성균주와 병원성균주로 처리된 담배 사이, 감수성품종인 BY4와 저항성품종 NC82 사이에도 두 효소의 활성과 동위효소형태의 차이가 없었다.

• 한국토양비료학회지
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• 제50권5호
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• pp.489-496
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• 2017

This study was conducted to test the applicability of hydroxy radical reactor system, which applied advanced oxidation processes, to sterilize pathogenic bacteria for nutrient solution recycling in closed hydroponics. Removal efficiency was tested on 25 L of nutrient solution maxed with 10 mL culture solution of bacteria, E. coli, and R. solanacearum in a pilot tank. The testing conditions included various levels of hydroxy radicals resulting from air flow rates of 40, 80, and $120L\;min^{-1}$, and 12 hours processing time. The removal of bacteria, E. coli, and R. solanacearum by hydroxy radical in nutrient solution was significantly increased with an increase in the flow rate of the air from $40L\;min^{-1}$ to $120L\;min^{-1}$. The optimum removal efficiency was achieved at an air flow rate of $120L\;min^{-1}$ for 2 hours treatment. There were no significant differences in removal efficiency among bacteria, E. coli, and R. solanacearum for tested level and time of hydroxy radical. These results verified the efficiency of hydroxy radical in removing the pathogenic bacteria and the applicability of hydroxy radical reactor system in the field.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.193-201
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• 2010

A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.