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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： Pan, Z. C.; Xu, J. 1 ; Prior, P.; Xu, J. S. 1 ; Zhang, H. 1 ; Chen, K. Y. 1 ; Tian, Q. 1 ; Zhang, L. Q. 1 ; Liu, L. 1 ; H 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

2.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Pla

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 ）

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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.

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