小麦TaFRA抗逆基因簇启动子的克隆及功能分析

In China, freshwater is very necessitous, so drought and salinity are two main abiotic stresses that affect the crop growth and production. With more and more new genes cloned, transgenic industrialization has developed rapidly. Promoter plays an important role in gene transcript, which will help to further understand the gene's function and mechanism, as well as to provide basis for biological safity of transgenic plant. In our previous studies, a new F-box gene related to abiotic stress was isolated, named as TaFRA, funded by National Natural Science Foundation of China (NSFC). There were three copies tendemly in wheat. Based on that, the present project will study their space-time expression model and different functions through transgenic rice with the three promoters of TaFRA gene cluster, and explore the specific element and transcript factors related to stress response by deleting some sequence from the promoters and yeast one hybrid. This was expected to provide help for identifing the gene's function and obtaining the inducible promoter originated from plant for wheat transgeic breeding.

我国淡水资源相对贫乏，干旱和盐碱胁迫严重地影响了粮食作物的生产，随着越来越多的控制重要性状的新基因分离克隆，植物转基因产业化步伐不断加快。启动子是基因转录中最主要的一种调节方式，加强植物源诱导型启动子研究对深入认识基因功能、机理具有辅助作用，而且对转基因生物安全具有潜在价值。本课题组在国家自然科学基金的资助下，从小麦耐盐品系98-160材料中克隆到与耐盐密切相关的新基因TaFRA，并有三个拷贝串联存在。本研究拟在前期工作基础上，通过对TaFRA基因簇三个拷贝启动子的水稻遗传转化分析，研究其时空表达模式及其分工，并试图通过缺失型启动子和酵母单杂交的方法来寻找可能的响应胁迫的特异元件和转录因子，以期对TaFRA基因簇表达模式并获得植物源诱导型启动子，为麦类作物转基因育种奠定基础。

植物的抗逆性与基因的转录调控密切相关，启动子是基因转录中最主要的一种调节方式，加强植物源诱导型启动子研究对深入认识基因功能、机理具有辅 助作用，而且对转基因生物安全具有潜在价值。本项目前期研究在小麦中发现耐逆相关基因TaFRA三个拷贝串联存在，这三个基因序列很保守，但在启动子区却存在较大差异。本研究将三个基因的启动子与GUS基因融合构建表达载体，转化水稻，对转基因水稻的GUS表达活性进行分析，发现三个启子对胁迫反应具有诱导活性，而且三个启动子的驱动活性在胁迫处理条件下存在差异，表明他们受到胁迫后具有一定的互补或增强效应；将不同缺失片段的启动子转化拟南芥，通过GUS表达活性分析，发现启动子的活性区域集中在1000bp以内，根据对启动子序列的顺式元件进行分析，确定了一些与非生物胁迫反应相关的核心区域，及能与其相互作用的转录因子，该结果为解析TaFRA基因串联重复的作用机制及基因的胁迫反应模式奠定了基础，同时获得了三个植物源非生物胁迫诱导型转录因子，对于麦类作物转基因研究具有一定的应用价值。