• Title/Summary/Keyword: Ralstonia solanacearum

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Detection of the Causal Agent of Bacterial Wilt, Ralstonia solanacearum in the Seeds of Solanaceae by PCR (가지과 종자에서 Ralstonia solanacearum의 검출을 위한 PCR 방법)

  • Cho, Jung-Hee;Yim, Kyu-Ock;Lee, Hyok-In;Baeg, Ji-Hyun;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.184-190
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    • 2011
  • Ralstonia solanacearum, a causal agent of bacterium wilt is very difficult to control once the disease becomes endemic. Thus, Ralstonia solanacearum is a plant quarantine bacterium in many countries including Korea. In this study, we developed PCR assays, which can detect Ralstonia solanacearum from the Solanaceae seeds. Primers RS-JH-F and RS-JH-R amplified specifically a 401 bp fragment only from Ralstonia solanacearum race 1 and race 3. The nested PCR primers, RS-JH-F-ne and RS-JH-R-ne that were designed inside of 1st PCR amplicon amplified specifically a 131 bp fragment only from Ralstonia solanacearum race 1 and race 3. The primers did not amplify any non-specific DNA from the seed extracts of the Solanaceae including tomato and pepper. When detection sensitivity were compared using the Solanaceae seeds inoculated with target bacteria artificially, the nested PCR method developed in this study 100 times more sensitive than ELISA and selective medium. Therefore, we believe that the PCR assays developed in this work is very useful to detect Ralstonia solanacearum in the Solanaceae seeds.

A Study on the Ralstonia Solanacearum Inactivation using Improved Plasma Process (개선된 플라즈마 공정을 이용한 Ralstonia Solanacearum 불활성화에 관한 연구)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Environmental Science International
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    • v.23 no.3
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    • pp.369-378
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    • 2014
  • Effect of improvement of the dielectric barrier discharge (DBD) plasma system on the inactivation performance of bacteria were investigated. The improvement of plasma reactor was performed by combination with the basic plasma reactor and UV process or combination with the basic plasma reactor and circulation system which was equipped with gas-liquid mixer. Experimental results showed that tailing effect was appeared after the exponential decrease in basic plasma reactor. There was no enhancement effect on the Ralstonia Solanacearum inactivation with combination of basic plasma process and UV process. The application of gas-liquid mixing device on the basic plasma reactor reduced inactivation time and led to complete sterilization. The effect existence of gas-liquid mixing device, voltage, air flow rate (1 ~ 5 L/min), water circulation rate (2.8 ~ 9.4 L/min) in gas-liquid mixing plasma, plasma voltage and UV power of gas-liquid mixing plasma+UV process were evaluated. The optimum air flow rate, water circulation rate, voltage of gas-liquid mixing system were 3 L/min, 3.5 L/min and 60 V, respectively. There was no enhancement effect on the Ralstonia Solanacearum inactivation with combination of gas-liquid mixing plasma and UV process.

Inactivation of Ralstonia Solanacearum using Filtration-Plasma Process (여과-Plasma 공정을 이용한 Ralstonia Solanacearum 불활성화)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Environmental Science International
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    • v.23 no.6
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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.

Detection of Ralstonia solanacearum with Nested PCR and DNA Enzyme-Linked Immunosorbent Assay (Nested PCR과 DNA Enzyme-Linked Immunosorbent Assays를 이용한 Ralstonia solanacearum의 검출)

  • Ko, Young-Jin;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.179-185
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    • 2007
  • In this study, we used the method of guanidin isothiocyanate and boiling with Chelex-100 resin to extract genomic DNA of Ralstonia solanacearum from soil. It is more efficient than general protocols to remove inhibitory compounds in soil and R. solanacearum on. Then, we applied polymerase chain reaction and DNA enzyme-linked immunosorbent assay (ELISA) to identify and detect pathogen. The fliC gene of R. solanacearum was selected for specific detection of pathogen and primer sets were designed. Among the primer sets, two specific and sensitive primer sets, RsolfliC(forward: 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J. $Sch\ddot{o}nfeld$ et al.) and RS_247 (forward: 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3 designed by this study), were designed to perform nested PCR. Nested PCR primer was labeled with biotin for hybridization between nested PCR product and probe to analyze with DNA ELISA.

Occurrence and Biovar Classification of Bacterial Wilt Caused by Ralstonia solanacearum in Eggplant (Solanum melongena) (가지의 Ralstonia solanacearum에 의한 풋마름병 발생과 생리형의 분화)

  • Lim, Yang-Sook;Lee, Mun-Jung;Cheung, Jong-Do;Rew, Young-Hyun;Kim, Byung-Soo
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.10-14
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    • 2008
  • Batcterial wilt caused by Ralstonia solanacearum is one of important and widespread diseases worldwide as well as in Korea. Bacterial wilt disease caused by R. solanacearum has been reported mainly in solanaceous crops including eggplant (Solanum melongena), tomato (Solanum lycopersicum), potato (S. tuberosum), and pepper (Capsicum annuum). A total of 48 strains of R. solanacearum from eggplant were collected during 2005 and 2006. They were confirmed as R. solanacearum by PCR amplification with primer pair flipcF/flipcR resulting in production of 470-bp DNA fragment. The 15 isolates exhibited pathogenicity on eggplant and tomato, but less virulent on pepper than other species. The biovar of collected isolates, which have been reported of five types worldwide, were classified as biovars 3 and 4 by physiological test. Biovar 4 was the dormant type without pathogenicity on eggplant rootstock, whereas biovar 3 had pathogenicity on eggplant rootstocks that is resistant to R. solanacearum, indicating necessity of breeding new rootstock with resistance to R. solanacearum biovar 3

Physiological, Biochemical and Genetic Characteristics of Ralstonia solanacearum Strains Isolated from Pepper Plants in Korea (고추에서 분리된 Ralstonia solanacearum 계통의 생리, 생화학 및 유전적 특성)

  • Lee, Young Kee;Kang, Hee Wan
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.265-272
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    • 2013
  • Totally sixty three bacteria were isolated from lower stems showing symptoms of bacterial wilt on pepper plants in 14 counties of 7 provinces, Korea. The isolates showed strong pathogenicity on red pepper (cv. Daewang) and tomato (cv. Seogwang) seedlings. All virulent bacteria were identified as Ralstonia solanacearum based on colony types, physiological and biochemical tests and polymerase chain reaction (PCR). All R. solanacearum isolates from peppers were race 1. The bacterial isolates consisted of biovar 3 (27%) and biovar 4 (73%). Based on polymorphic PCR bands generated by repetitive sequence (rep-PCR), the 63 R. solanacearum isolates were divided into 12 groups at 70% similarity level. These results will be used as basic materials for resistant breeding program and efficient control against bacterial wilt disease of pepper.

First Report of Bacterial Wilt Caused by Ralstonia solanacearum Biovar 2 Race 1 on Tomato in Egypt

  • Seleim, Mohamed A.A.;Abo-Elyousr, Kamal A.M.;Abd-El-Moneem, Kenawy M.;Saead, Farag A.
    • The Plant Pathology Journal
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    • v.30 no.3
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    • pp.299-303
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    • 2014
  • This study aims to isolate and identify the causal pathogen of tomato bacterial wilt in Egypt. In 2008, tomato plants showing typical symptoms of bacterial wilt disease with no foliar yellowing were observed in Minia, Assiut and Sohag governorates, Egypt. When cut stems of symptomatic plants were submerged in water, whitish ooze was evident and longitudinal sections showed a brown discoloration in the vascular tissues. Bacteria were isolated on triphenyl tetrazolium chloride medium and fifteen isolates shown typical morphological and cultural characteristics were confirmed as Ralstonia solanacearum biovar 2 race 1. Pathogenicity tests showed that all isolates proved to be pathogenic to tomato plants, varied from 52 to 97% wilting. This is the first report of R. solanacearum biovar 2 race 1 causing bacterial wilt in tomato crop in Egypt.

Improvement of Biological Control against Bacterial Wilt by the Combination of Biocontrol Agents with Different Mechanisms of Action

  • Kim, Ji-Tae;Kim, Shin-Duk
    • Journal of Applied Biological Chemistry
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    • v.50 no.3
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    • pp.136-143
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    • 2007
  • Despite the increased interests in biological control of soilborne diesease for environmental protection, biological control of bacterial wilt caused by Ralstonia solanacearum have not provided consistent or satisfying results. To enhance the control efficacy and reducing the inconsistency and variability, combinations of specific strains of microorganisms, each having a specific mechanism of control, were applied in this study. More than 30 microorganisms able to reduce the activity of pathogen by specific mechanism of action were identified and tested for their disease suppressive effects. After in vitro compatibility examinations, 21 individual strains and 15 combinations were tested in the greenhouse. Results indicated three-way combinations of different mode of control, TS3-7+A253-16+SKU78 and TS1-5+A100-1+SKU78, enhanced disease suppression by 70%, as compared to 30-50% reduction for their individual treatments. This work suggests that combining multiple traits antagonizing the pathogen improve efficacy of the biocontrol agents against Ralstonia solanacearum.

Characterization of an Antibiotic Produced by Bacillus subtilis JW-1 that Suppresses Ralstonia solanacearum

  • Kwon, Jae Won;Kim, Shin Duk
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.13-18
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    • 2014
  • Bacillus subtilis JW-1 was isolated from rhizosphere soil as a potential biocontrol agent of bacterial wilt caused by Ralstonia solanacearum. Seed treatment followed by a soil drench application with this strain resulted in >80% reduction in bacterial wilt disease compared with that in the untreated control under greenhouse conditions. The antibacterial compound produced by strain JW-1 was purified by bioactivity-guided fractionation. Based on mass spectroscopy and nuclear magnetic resonance spectral data ($^1H$, $^{13}C$, $^1H-^1H$ correlation spectroscopies, rotating frame nuclear Overhauser effect spectroscopy, and heteronuclear multiple-bond correlation spectroscopy), the structure of this compound was elucidated as a cyclic lipopeptide composed of a heptapeptide (Gln-Leu-Leu-Val-Asp-Leu-Leu) bonded to a ${\beta}$-hydroxy-iso-hexadecanoic acid arranged in a lactone ring system.