title: Loop-Mediated Isothermal Amplification Method for the Rapid Detection ofRalstonia solanacearumPhylotype I Mulberry Strains in China.
abstract: Ralstonia solanacearumphylotype I mulberry strains are causative agent of bacterial wilt of mulberry. Current diagnostic methods are not adopted to the mulberry wilt disease. In this study, we developed a rapid method, loop-mediated isothermal amplification (LAMP), to detectR. solanacearumphylotype I mulberry strains. A set of six primers was designed to target the clone MG67 sequence in this LAMP detection which can be completed in 20 min at 64°C. The results of the LAMP reaction could be observed with the naked eye due to magnesium pyrophosphate precipitate produced during the reaction or the color change after adding SYBR Green I. The specificity of the LAMP was confirmed using DNA from 46 representative strains ofR. solanacearumand 7 other soil-borne bacteria strains. This method was also of high sensitivity and could be used to detect the presence of less than 160 fg genomic DNA or 2.2 × 102CFU/ml of bacterial cells per 25 μl reaction volume, moreover, the presence of plant tissue fluid did not affect the sensitivity. Since it does not require expensive equipment or specialized techniques, this LAMP-based diagnostic method has the potential to be used under field conditions to make disease forecasting more accurate and efficient.
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural SciencesBeijing, China; College of Plant Protection, Southwest UniversityChongqing, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Huang
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Jingsheng
authors.initials: J
authors.lastname: Xu
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Xiangjiu
authors.initials: X
authors.lastname: Kong
authors.affiliation: College of Plant Protection, Southwest University Chongqing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ding
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Feng
keywords: Ralstonia solanacearum; bacterial wilt; detection; loop-mediated isothermal amplification; phylotype I mulberry strains