PpADC基因调控桃树抗逆性的分子机制研究

项目名称： PpADC基因调控桃树抗逆性的分子机制研究

项目编号： No.31501727

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 王保全

作者单位： 河南科技学院

项目金额： 20万元

中文摘要： 前期研究表明基因PpADC（桃树精氨酸脱羧酶）是一个多重胁迫应答基因，其转录表达明显受到干旱、低温和高盐等胁迫诱导。分离PpADC启动子进行生物信息学分析表明，PpADC启动子上含有干旱、低温等逆境应答顺式作用元件。在此基础上，本项目首先比较逆境处理下的植株表型，测定相关生理指标，以验证PpADC的抗逆功能。构建启动子不同类型的缺失载体转化Micro-Tom番茄，借助GUS染色和荧光定量技术，鉴定启动子的逆境调控功能，通过凝胶阻滞和定点突变分析确定启动子上逆境应答顺式作用元件，酵母单杂交筛选与逆境顺式作用元件特异结合的转录因子，结合转录因子的亚细胞定位及其编码基因的逆境转录表达模式，解析基因PpADC调控桃树抗逆性的分子机制。本项目是已有研究的深入和延续，既为揭示PpADC应答逆境的转录调控机制奠定基础，又为桃树抗逆遗传改良工程发掘优良基因资源，具有重要理论意义和潜在应用价值。

中文关键词： 桃；精氨酸脱羧酶；抗逆性；启动子；转录因子

英文摘要： Our previous work showed that PpADC was obviously induced by drought, low temperature and salt stresses, suggesting that PpADC was responsive to various stresses. The promoter of PpADC was isolated and bioinformatics analysis indicated that it contained the putative cis-elements related to drought, low temperature and so on. Based on the obtained data, this project first compares the differences of the phenotype between the transgenic lines and WT, and then related physiological indexes will be measured, including electrolyte leakage and chlorophyll content. Promoter deletions will be conducted and the corresponding vectors will be transferred into Micro-Tom tomato. GUS staining and fluorescent quantification will be used to identify the function of the promoter in response to stresses. Cis-elements on the promoter will be analyzed via electrophoretic mobility shift assay (EMSA) and site-directed mutagenesis. Then the corresponding transcription factors binding to the cis-element will be selected by yeast one-hybrid. In addition, subcellular localization and transcriptional changes responsive to stresses will be analyzed. This project is based on previously obtained data, it will have in-depth analysis on characterization of the function of PpADC and provide candidate gene for genetic improvement of peach.

英文关键词： prunus persica;ADC;stress resistance;promoter;transcription factor