巨桉锌指结构蛋白基因EgrZPCT在抗冷胁迫中功能和调控机制的研究

项目名称： 巨桉锌指结构蛋白基因EgrZPCT在抗冷胁迫中功能和调控机制的研究

项目编号： No.31270657

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 程龙军

作者单位： 浙江农林大学

项目金额： 78万元

中文摘要： 研究巨桉抗冻的分子机制，对扩大其栽培领域，降低低温造成的经济损失有重要意义。我们从巨桉不同低温处理的数字表达谱（DGE）数据中，得到一个受低温强烈诱导的编码锌指结构蛋白的基因EgrZPCT，并对其基因和启动子序列进行了克隆。前期研究表明，该基因受低温特异诱导，蛋白质与拟南芥ZAT12同源性较高，很可能是参与巨桉抗冷一条重要分子途径中的关键基因。本课题准备构建超表达、干涉和启动子指导GUS表达载体进行巨桉的遗传转化，通过对转基因植株在不同低温处理条件下的分子、生理分析，明确EgrZPCT的功能；另一方面，由于植物抗冷分子机制中，锌指蛋白类基因的调控机制至今不明确，我们准备运用酵母单杂交和CASTing技术，对调控EgrZPCT表达的上游基因和EgrZPCT所调控下游基因启动子上的结合序列，进行钓取和筛选，明确EgrZPCT在巨桉抗冷分子途径中的调控机制。

中文关键词： 巨桉；锌指蛋白；抗冷；功能；调控机制

英文摘要： The planting area of Eucalyptus grandis tree will be expanded and the lost caused by low temperture also will be decreased, if we learned about the molecular mechanism of cold tolerance in it. According to our data of digital gene expression profiling (DGE) under the condition of different low temperature treatment to Ecucalyputs grandis seedlings, a gene which encodes zinc-finger protein was selected because its expression was induced strongly under low temperature. Result of our previous research showed that the protein sequence of it has a high similarity to AtZAT12, a gene playing an important role in a special pathway related with cold tolerance. And, the quantitative RT-PCR result showed the gene was only induced by low temperature, and other abiotic stresses has no influence on its expression. We predicted that the gene was involed in the cold tolerance of Eucalyptus grandis and named it EgrZPCT(zin-finger protein of cold tolerance).Its full length of cDNA sequence and promotor sequence has been cloned. To elucidate the function of EgrZPC, overexpression, RNA interference and promotor/GUS vectors will be constructed and tranformed to Eucalyptus. The transgene seedlings will be used to analyze the function of EgrZPCT with molecular and physiology experiments. The research on regulation mechanisms of EgrZP

英文关键词： Eucalyptus grandis；zinc finger protein；cold tolerance；function；regulation mechanism