水稻PBZ1基因启动子对烯丙异噻唑的诱导响应性分析  

Analysis of the responsiveness of rice PBZ1 promoter to probenazole

作　　者：朱政[1,2] 王迟早 高炯[3] 丁雨龙[1,4] 蒯本科[3] 

机构地区：[1]南京林业大学南方现代林业协同创新中心,江苏南京210037 [2]南京林业大学生物与环境学院,江苏南京210037 [3]复旦大学生命科学学院/遗传工程国家重点实验室,上海200438 [4]南京林业大学竹类研究所,江苏南京210037

出　　处：《南京农业大学学报》2015年第5期720-726,共7页Journal of Nanjing Agricultural University

基　　金：国家转基因生物新品种培育重大专项(2013ZX08009-004);江苏省优势学科建设项目

摘　　要：[目的]依据水稻PBZ1基因对一种环境友好型化学诱导剂烯丙异噻唑(3-allyloxy-1,2-benzisothiazole-1,1-dioxide,PBZ)的高响应性,旨在分析该基因启动子作为一种新型化学诱导启动子的应用潜力。[方法]首先构建PBZ1启动子与GUS的融合报告载体PPBZ1-1942bp::GUS,分别采用烟草叶片瞬时表达及转基因水稻稳定表达系统,检测PBZ1启动子对PBZ的诱导响应性;并且利用水稻原生质体瞬时表达系统,进一步分析不同长度的PBZ1启动子片段的诱导响应性。[结果]无论是在瞬时还是稳定表达系统中检测,PBZ1启动子均可以有效响应PBZ的诱导。本研究还首次证明水稻PBZ1基因响应PBZ诱导很大程度上依赖于茉莉酸(JA)信号途径。因此,在水稻原生质体瞬时表达系统中,我们采用茉莉酸甲酯(Me JA)作为诱导剂,替代不适用于水稻原生质体体系的PBZ,有效地验证了1 053和608 bp长度的启动子片段具有更好的诱导响应性。[结论]水稻PBZ1启动子具备作为一种基于PBZ诱导系统的新型化学诱导启动子的潜力。[Objectives]The expression of rice PBZ1 gene is highly induced by probenazole（ 3-allyloxy-1,2-benzisothiazole-1,1-dioxide,PBZ）,a widely used and environment-friendly chemical for rice blast control. This study aims to analyze the application potential of PBZ1 promoter as a new chemical-inducible promoter. [Methods]In this study,the PBZ1 promoter was fused with GUS reporter gene,resulting in the recombinant construct PPBZ1-1942bp： ： GUS. Then,the responsiveness of PBZ1 promoter to PBZ was tested by transient expression in tobacco leaves and by stable expression in transgenic rice. We also analyzed the inducible expression activity of different PBZ1 promoter fragments by transient expression in rice protoplast. [Results]Both in the transient expression system and the stable expression system,the results showed that the PBZ1 promoter can effectively respond to PBZ. In this research,we also found that the induction of PBZ1 to PBZ mainly depended on the jasmonic acid（ JA） signaling pathway. Thus,methyl jasmonate（ Me JA）,instead of PBZ,was used as the inducer of PBZ1 in rice protoplast. The 1 053 bp and 608 bp promoter fragments of PBZ1 showed higher inducibility to Me JA than the other two fragments in the transient expression system of rice protoplast. [Conclusions]The promoter of PBZ1 is an ideal candidate for engineering a new chemical-inducible expression system.

关 键 词：烯丙异噻唑 PBZ1 化学诱导剂 化学诱导启动子 

分 类 号：S511.032[农业科学—作物学]