河南农业科学2017，Vol.46Issue(4)：80-83,94,5.DOI:10.15933/j.cnki.1004-3268.2017.04.015
烟草青枯菌Tn5突变体库的构建及突变位点分析
Construction of Tn5 Mutant Library of Ralstonia solanacearum and Analysis of Mutation Sites
摘要
Abstract
In order to explore the pathogenic genes from the whole genome range of Ralstonia solanacearum,this study constructed a Ez-Tn5 transposon insertion mutant library of R.solanacearum TX-LLJ14-3 by electroporation method.The library contained 12 000 mutants.There were 216 non or weak pathogenic mutant strains by the pathogenic detection on tobacco.Tn5 flanking sequences of 15 non pathogenic mutant strains were amplified by TAIL-PCR,and simultaneously,the insertion sites were analyzed.The results showed that the insertion sites of the 15 mutant strains were located at genes with different functions such as nucleotide hydrolases,glycosyltransferases,transposase and synthase.When they were disrupted or destroyed,the expression or secretion of virulence related substances might be inhibited,or tobacco resistance to pathogen be induced,thus showing no pathogenic characteristics.关键词
烟草青枯菌/Tn5插入突变/无致病力菌株/TAIL-PCR/突变位点Key words
Ralstonia solanacearum/Tn5 insertion mutation/non pathogenic strain/TAIL-PCR/mutation site分类
农业科技引用本文复制引用
李小杰,李淑君,李成军,陈玉国,王海涛,邱睿,胡亚静..烟草青枯菌Tn5突变体库的构建及突变位点分析[J].河南农业科学,2017,46(4):80-83,94,5.基金项目
河南省基础与前沿技术研究计划项目(152300410142) （152300410142）
河南省烟草公司科技项目(HYKJ201407) （HYKJ201407）
