数字农科院2.0

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

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

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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