重金属镉Cd胁迫反应中转录因子干扰肽siPEP的功能鉴定

项目名称： 重金属镉Cd胁迫反应中转录因子干扰肽siPEP的功能鉴定

项目编号： No.31271421

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 谭明普

作者单位： 南京农业大学

项目金额： 70万元

中文摘要： 本项目拟以RNA-seq分析镉Cd胁迫水稻的转录组为出发点，结合水稻基因组注释，确定与转录因子同源的小分子干扰肽siPEP。在转录组测序得到差异表达基因的基础上，以转录因子为核心，利用酵母双杂交和双分子荧光互补系统BiFC等蛋白质互作分析方法，筛选与转录因子互作的siPEPs；采用酵母单杂交分析和原生质体转录激活活性分析体系，确证转录因子与其靶调节的调节关系。初步明确siPEP干扰哪些转录因子来间接调节胁迫反应基因，进一步通过转基因来验证以上推测的作用模式。预期结果不仅可以从一个全新的角度揭示植物胁迫反应机制和重金属耐受机理，而且可以为植物遗传改良提供新的基因资源，对于环境修复和粮食安全具有重要的指导意义。

中文关键词： 小分子干扰肽；NAC转录因子；γ氨基丁酸；耐镉基因；全基因组关联分析

英文摘要： In this project, the analysis of the transcriptome of rice under cadmium stress as a starting point, combined with the rice genome annotation, the small interfering peptides, designated siPEPs, homologous to transcription factor, will be identified. Also, the differentially expressed genes will be obtained from the comparative transcriptome sequencing. Then, yeast two-hybrid screening and bimolecular fluorescence complementation (BiFC) system are used to confirm the interaction between siPEP and transcription factor. Yeast one-hybrid and protoplast-based transcriptional activation analysis systems are utilized to verify the target gene of the transcription factor. Thus, the mechanism of siPEP regulating the stress response through interfering certain transcription factors will be clarified. This proposed model centered on transcription factor will be verified by the transgenic research focusing on siPEP regulated Cd stress response. The expected results are not only helpful to reveal the mechanism of plant stress response mechanisms and heavy metal tolerance from a new perspective, but also provide new genetic resources for plant genetic improvement. whether in theory or in application, all this results have important guiding significance for environmental rehabilitation and food security.

英文关键词： Small interfering peptides；NAC transcription factors；gamma-aminobutyric acid；Cadmium tolerance gene；whole genome-wide association analysis