水稻细菌性条斑病菌对噻唑锌抗药性相关基因的克隆及其功能研究

项目名称： 水稻细菌性条斑病菌对噻唑锌抗药性相关基因的克隆及其功能研究

项目编号： No.31201550

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 陈雨

作者单位： 安徽省农业科学院

项目金额： 24万元

中文摘要： 噻唑锌是我国自主开发的一种新型杀细菌剂，对水稻细菌性条斑病菌具有较高的抑制活性，但抗药性机制并不是很清楚。为克隆抗药性相关基因，本项目研究内容为：（1）通过构建抗噻唑锌的水稻细菌性条斑病菌的转座子插入突变基因文库，以丧失抗药性遗传表型为指标筛选转化子；（2）通过Tail-PCR扩增被Tn5转座子插入而破坏的基因（简称为抗药性相关基因）的侧翼序列，根据NCBI的信息获得该基因的全序列；（3）扩增抗、感菌株中的抗药性相关基因，经比对、分析，明确该基因与病原菌抗药性之间的关系；（4）构建敲除突变体及回复突变体，通过PCR及杂交验证抗药性基因，明确突变体的生物学性质，揭示该基因的生物学功能及其与致病性的关系，从而为探明水稻细菌性条斑病菌对噻唑锌的抗药性分子机制奠定基础。其结果不仅能够为噻唑锌的抗药性风险评估提供重要的参考依据，而且为该药剂的科学使用、抗药性监测以及延缓抗药性的产生提供理论基础。

中文关键词： 水稻细菌性条斑病菌；噻唑锌；抗药性相关基因；基因功能；

英文摘要： Zinc thiazole is a newly independently developed bactericide in China, which exhibited strong inhibitory activity against Xanthomonas oryzae pv. oryzicola. But the mechanism of resistance to zinc thiazole is still unclearly understood. In order to clone the resistance-related gene(s), the objctives of this program are to: (1) established the Tn5 insertional mutants library from the zinc thiazole-resistant X. oryzae pv. oryzicola strain and the identify the transformants which lost the resistance to zinc thiazole; (2) amplify the flanking sequences of the Tn5-inserted gene through tail-PCR and get the whole sequence of the gene (resistance-related gene) based on the information through NCBI;(3) amplify the whole gene from the zinc thiazole-sensitive and -resistant X.oryzae pv. oryzicola strains with sequencing and blasting, so as to investigate the relationship between the gene and the resistance; (4) creat the in-frame deleted mutants and the complementary mutants, identify the gene function through PCR and blots, explore the characters of the mutants and the biological properties of the resistance-related gene and its relationship with pathogenicity. This would make basis for the molecular mechanism of resistance to zinc thiazole in X. oryzae pv. oryzicola. The results of this program would not only provide imp

英文关键词： Xanthomonas oryzae pv. oryzicola；Zinc thiazole；resistance-related gene；gene function；