Development of a specific molecular tool for the detection of epidemiologically active mulberry causing-disease strains of Ralstonia solanacearum phylotype I (historically race 5-biovar 5) in China

文献类型: 外文期刊

第一作者: Xu, J.

作者: Xu, J.;Xu, J. S.;Zhang, H.;Chen, K. Y.;Tian, Q.;Zhang, L. Q.;Liu, L.;He, L. Y.;Feng, J.;Prior, P.;Pan, Z. C.

作者机构:

关键词: Mulberry;Ralstonia solanacearum;Detection;Suppression subtractive hybridization;Insertion sequences

期刊名称:EUROPEAN JOURNAL OF PLANT PATHOLOGY ( 影响因子:1.907; 五年影响因子:2.022 )

ISSN:

年卷期:

页码:

收录情况: SCI

摘要: Ralstonia solanacearum causes bacterial wilt disease in many plant species, including mulberry. Here, we used a suppression subtractive hybridization (SSH) approach to identify specific DNA fragments in R. solanacearum race 5-biovar 5. The genome of the R. solanacearum M7 strain was subtracted from that of the GMI1000 strain, resulting in the identification of 85 subtracted fragments. The primer set MG67-F/R for identification of Ralstonia solanacearum race 5-biovar 5 strains was designed on the basis of the clone MG67 sequence. Furthermore, a multiplex PCR was developed by using the primer set MG67-F/MG67R in combination with the species-specific primer pair 759/760. A 156 bp r5-bv5-specific fragment, together with a 282 bp species-specific fragment, was amplified from all tested R. solanacearum r5-bv5 strains. The sensitivity of the multiplex PCR made it possible to detect concentrations as low as 10(2) CFU ml(-1) of pure culture. Moreover, the r5-bv5-specific multiplex PCR was successfully applied to detect Ralstonia solanacearum race 5-biovar 5 strains in diseased mulberry samples. Therefore, the multiplex PCR assay can be used as a reliable diagnostic technique to enable researchers to rapidly identify isolates of R. solanacearum race 5-biovar 5.

分类号: S432.1

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