• Title/Summary/Keyword: Ralstonia Solanacearum

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Mutation of Virulence Candidate Genes of Ralstonia solanacearum

  • Park, Eun-Jin;Lee, Seung-Don;Um, Hae-Young;Chung, Eu-Jin;Lee, Myung-Hwan;Moon, Byung-Ju;Lee, Seon-Woo
    • 한국생명과학회:학술대회논문집
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    • 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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    • pp.104.1-104.1
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    • 2007
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A Real-Time PCR Assay for the Quantitative Detection of Ralstonia solanacearum in Horticultural Soil and Plant Tissues

  • Chen, Yun;Zhang, Wen-Zhi;Liu, Xin;Ma, Zhong-Hua;Li, Bo;Allen, Caitilyn;Guo, Jian-Hua
    • 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.

Identification of a Gene Involved in the Negative Regulation of Pyomelanin Production in Ralstonia solanacearum

  • Ahmad, Shabir;Lee, Seung Yeup;Khan, Raees;Kong, Hyun Gi;Son, Geun Ju;Roy, Nazish;Choi, Kihyuck;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • 제27권9호
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    • pp.1692-1700
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    • 2017
  • Ralstonia solanacearum causes bacterial wilt in a wide variety of host plant species and produces a melanin-like blackish-brown pigment in stationary phase when grown in minimal medium supplemented with tyrosine. To study melanin production regulation in R. solanacearum, five mutants exhibiting overproduction of melanin-like pigments were selected from a transposon (Tn) insertion mutant library of R. solanacearum SL341. Most of the mutants, except one (SL341T), were not complemented by the original gene or overproduced melanins. SL341T showed Tn insertion in a gene containing a conserved domain of eukaryotic transcription factor. The gene was annotated as a hypothetical protein, given its weak similarity to any known proteins. Upon complementation with its original gene, the mutant strains reverted to their wild-type phenotype. SL341T produced 3-folds more melanin at 72 h post-incubation compared with wild-type SL341 when grown in minimal medium supplemented with tyrosine. The chemical analysis of SL341T cultural filtrate revealed the accumulation of a higher amount of homogentisate, a major precursor of pyomelanin, and a lower amount of dihydroxyphenylalanine, an intermediate of eumelanin, compared with SL341. The expression study showed a relatively higher expression of hppD (encoding hydroxyphenylpyruvate dioxygenase) and lower expression of hmgA (encoding homogentisate dioxygenase) and nagL (encoding maleylacetoacetate isomerase) in SL341T than in SL341. SL341 showed a significantly higher expression of tyrosinase gene compared with SL341T at 48 h post-incubation. These results indicated that R. solanacearum produced both pyomelanin and eumelanin, and the novel hypothetical protein is involved in the negative regulation of melanin production.

풋마름병균, Ralstonia solanacearum의 길항세균 SKU-78 균주의 분리 동정 및 특성 (Isolation, Identification and Biological Control Activity of SKU-78 Strain against Ralstonia solanacearum)

  • 성필제;신정균;조홍범;김신덕
    • Applied Biological Chemistry
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    • 제48권1호
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    • pp.48-52
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    • 2005
  • 종자발아실험과 뿌리에서의 집락 형성실험으로 고추와 토마토의 근권 토양에서 plant growth promoting rhizobacteria(PGPR) 균주를 선발하였다. 그 중 포트실험에서 종자처리와 토양 관주처리 하였을 때 60% 이상 풋마름병 방제효과를 나타낸 SKU-78 균주에 대해서 포장에서의 방제효과 검증에 의해 생물농약으로의 개발 가능성을 확인하였으며, 생화학적 특성조사와 16S rDNA sequence 분석에 의해 SKU-78 균주를 Bacillus sp. SKU-78로 동정하였다.

Plant-derived Antibacterial Metabolites Suppressing Tomato Bacterial Wilt Caused by Ralstonia solanacearum

  • Vu, Thuy Thu;Choi, Gyung Ja;Kim, Jin-Cheol
    • 식물병연구
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    • 제23권2호
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    • pp.89-98
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    • 2017
  • Ralstonia solanacearum species complex (RSSC) causes bacterial wilt, and it is one of the most important soil-borne plant pathogenic bacteria. RSSC has a large host range of more than 50 botanical families, which represent more than 200 plant species, including tomato. It is difficult to control bacterial wilt due to following reasons: the bacterial wilt pathogen can grow inside the plant tissue, and it can also survive in soil for a long period; moreover, it has a wide host range and biological diversity. In most previous studies, scientists have focused on developing biological control agents, such as antagonistic microorganisms and botanical materials. However, biocontrol attempts are not successful. Plant-derived metabolites and extracts have been promising candidates to environmentally friendly control bacterial wilt diseases. Therefore, we review the plant extracts, essential oils, and secondary metabolites that show potent in vivo antibacterial activities (in potted plants or in field) against tomato bacterial wilt, which is caused by RSSC.

연초의 세균성마름병 저항성 유전에 관한 연구 (Studies on the Inheritance of Resistance to Bacterial Wilt(Ralstonia solanacearum) in Tobacco(Nicotiana tabacum L.))

  • 정석훈
    • 한국연초학회지
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    • 제22권1호
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    • pp.25-30
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    • 2000
  • Bacterial wilt(Ralstonia solanacearum) is one of the major diseases of flue-cured tobacco (Nicotiana tabacum L.) in the world. This study was conducted to investigate degree of dominance, selection, and correlation between leaf shape and degree of bacterial wilt resistance in flue-cured tobacco. The degree of disease caused by bacterial wilt was evaluated in parents, F$_1$, F$_2$ and F$_3$ populations of two crosses, BY 4 x NC 95 and BY 4 x Coker 86, in the infected field. The leaf shape index was also measured in parents and F$_2$ population of BY 4 x NC 95. The incidence of bacterial wilt was observed in the middle of June and peaked in late July, when the highest value of pathogen density reached 1.0 x 10$^{6}$ colony forming unit per gram. It was concluded that the inheritance mode of risestance to bacterial wilt in the above two crosses of susceptible and resistant varieties was recessive and polygenic. The resistance to bacterial wilt was significantly correlated with leaf shape in F2 generation of BY 4 x NC 95. But certain plants having narrower leaves were also resistant to bacterial wilt. It is considered that the bacterial wilt resistant lines having narrower leaves could be selected. The selection for bacterial wilt resistance in the F$_2$ population might be effective.

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Bacillus subtilis JW-1 균주가 생산하는 bacilysin의 풋마름병 억제 효과 및 특성 (Isolation and characterization of bacilysin against Ralstonia solanacearum from Bacillus subtilis JW-1)

  • 김신덕
    • 미생물학회지
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    • 제54권2호
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    • pp.136-139
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    • 2018
  • 풋마름병 균에 대해 강한 저해활성을 갖는 활성물질 Compound S와 conversion product인 compound S'이 Bacillus subtilis JW-1 배양액에서 일련의 크로마토그래피 방법으로 분리 정제되었고, $^1H$ NMR, $^{13}C$ NMR, $^1H-^1H$ COSY와 HMBC 등의 spectra 분석에 의해 구조가 alanyl-L-${\beta}$-(2,3-epoxycyclohexyl-4-one)alanine와 alanyl-L-${\beta}$-(2,3-dihydroxycyclohexyl-4-one) alanine로 각각 동정되었다. Compound S는 $G^+$, $G^-$ bacteria, yeast와 Candida albicans 등에 대해 광범위한 antimicrobial activity를 나타내며, conversion product의 활성 상실을 통해 intact oxirane ring이 Compound S의 활성에 필수적임이 밝혀졌다.

한국에서 분리한 Ralstonia solanacearum에 대한 순계 토마토의 병 반응과 고온에서의 발병 (Disease Responses of Tomato Pure Lines Against Ralstonia solanacearum Strains from Korea and Susceptibility at High Temperature)

  • 이형주;조은정;김남희;채영;이선우
    • 식물병연구
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    • 제17권3호
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    • pp.326-333
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    • 2011
  • Ralstonia solanacearum에 의해 발생하는 토마토 풋마름병에 대하여 저항성으로 알려진 토마토 순계 품종의 국내 병원균에 대한 저항성 반응을 온도별로 평가하였다. 한국에서 분리된 R. solanacearum 균주로 토마토 순계 6 품종의 저항성을 평가한 결과 기존에 저항성으로 알려진 토마토 순계들이 한국 분리균주에 대하여는 대부분 저항성을 유지하지 못하였다. 병원성이 강한 균주인 SL341 (race 1, biovar 4) 균주는 검정한 대부분의 품종에서 온도에 관계없이 강한 병원성을 보였다. 반면, 담배에서 분리된 균주인 SL1944(race 1, biovar 4)은 온도에 따라 발병진전이 현저하게 차이가 났다. Moneymaker와 Bonny Best와 같은 품종은 온도에 관계없이 SL1944에 대하여 감수성이었다. 그러나, 풋마름병 저항성 품종으로 알려진 Hawaii 7998, Hawaii 7996, B-blocking 품종은 오히려 상대적으로 고온인 조건($35^{\circ}C$에서 14시간 명조건과 $28^{\circ}C$에서 10시간 암조건)에서 급격히 발병하였다. 병 진전은 동일한 품종의 낮은 온도에서 병 진전이나 Moneymaker나 Bonny Best 같은 품종에서 동일한 고온 조건의 발병에 비해 눈에 띄게 빨랐다. 본 연구결과는 국내에서 분리된 균주들이 기존에 저항성 토마토 품종을 가해할 수 있으며 고온조건에서는 품종의 저항성 붕괴의 가능성으로 급격한 풋마름병이 유발될 수 있음을 제시한다.

담배세균성마름병[립고병(立枯病)]에 대한 담배품종의 저항성 검정법 (Screening Procedure of Tobacco Cultivars for Resistant to Bacterial Wilt Caused by Ralstonia solanacearum)

  • 전용호;강여규
    • 한국연초학회지
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    • 제30권1호
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    • pp.1-7
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    • 2008
  • Bacterial wilt caused by Ralstonia solanacearum has become a severe problem on tobacco in Korea. No effective single control measure is available at present time. One of the most potential way for controlling the bacterial wilt on tobacco is growing tobacco cultivars resistant to the bacterial wilt. In this study, optimal conditions for screening tobacco cultivars resistant to the bacterial wilt were examined to provide reproducible and efficient methods in growth chamber testing and field experiments for evaluating plant disease resistance. For this, already-known inoculation methods, inoculum densities, and incubation temperature, and plant growth stages at the time of inoculation were compared using tobacco cultivars resistant (Nicotiana tabacum cv, NC95), moderately resistant (N. tabacum cv. SPG70), and susceptible (N. tabacum BY4) to the bacterial disease. It was determined that root-dipping of tobacco seedlings at six true leaf stage into the bacterial suspension with inoculum level of $10^8$ colony-forming units (CFU)/ml for 20 min before transplanting was simple and most efficient in testing for resistance to the bacterial wilt of tobacco caused by R. solanacearum, for which disease incidences and severities were examined at 2 weeks of plant growth after inoculation at $20{\sim}25^{\circ}C$ in a growth chamber. These experimental conditions could discriminate one tobacco cultivar from the others by disease severity better than any other experimental conditions. In field testing, the optimum time for examining the disease occurrence was late June through early July. These results can be applied to establishing a technical manual for the screening of resistant tobacco cultivars against the bacterial wilt caused by R. solanacearum.

Contribution of the murI Gene Encoding Glutamate Racemase in the Motility and Virulence of Ralstonia solanacearum

  • Choi, Kihyuck;Son, Geun Ju;Ahmad, Shabir;Lee, Seung Yeup;Lee, Hyoung Ju;Lee, Seon-Woo
    • The Plant Pathology Journal
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    • 제36권4호
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    • pp.355-363
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    • 2020
  • Bacterial traits for virulence of Ralstonia solanacearum causing lethal wilt in plants were extensively studied but are not yet fully understood. Other than the known virulence factors of Ralstonia solanacearum, this study aimed to identify the novel gene(s) contributing to bacterial virulence of R. solanacearum. Among the transposon-inserted mutants that were previously generated, we selected mutant SL341F12 strain produced exopolysaccharide equivalent to wild type strain but showed reduced virulence compared to wild type. In this mutant, a transposon was found to disrupt the murI gene encoding glutamate racemase which converts L-glutamate to D-glutamate. SL341F12 lost its motility, and its virulence in the tomato plant was markedly diminished compared to that of the wild type. The altered phenotypes of SL341F12 were restored by introducing a full-length murI gene. The expression of genes required for flagella assembly was significantly reduced in SL341F12 compared to that of the wild type or complemented strain, indicating that the loss of bacterial motility in the mutant was due to reduced flagella assembly. A dramatic reduction of the mutant population compared to its wild type was apparent in planta (i.e., root) than its wild type but not in soil and rhizosphere. This may contribute to the impaired virulence in the mutant strain. Accordingly, we concluded that murI in R. solanacearum may be involved in controlling flagella assembly and consequently, the mutation affects bacterial motility and virulence.