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

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 102 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.

Bibliographic Details

Main Authors:	Pan, Z.C., Xu, Jianchu, Prior, Philippe, Xu, J.S., Zhang, H., Chen, K.Y., Tian, Q., Zhang, L.Q., Liu, L., He, L.Y., Feng, J.
Format:	article biblioteca
Language:	eng
Subjects:	H20 - Maladies des plantes, Ralstonia solanacearum, biologie moléculaire, identification, hybridation, épidémiologie, technique analytique, PCR, Broussonetia papyrifera, http://aims.fao.org/aos/agrovoc/c_37076, http://aims.fao.org/aos/agrovoc/c_4891, http://aims.fao.org/aos/agrovoc/c_3791, http://aims.fao.org/aos/agrovoc/c_3706, http://aims.fao.org/aos/agrovoc/c_2615, http://aims.fao.org/aos/agrovoc/c_1513, http://aims.fao.org/aos/agrovoc/c_34079, http://aims.fao.org/aos/agrovoc/c_1117, http://aims.fao.org/aos/agrovoc/c_1556,
Online Access:	http://agritrop.cirad.fr/587410/ http://agritrop.cirad.fr/587410/1/s10658-013-0249-9.pdf
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