非ABA依赖型SnRK2激酶调控马铃薯响应干旱胁迫的机制解析

项目名称： 非ABA依赖型SnRK2激酶调控马铃薯响应干旱胁迫的机制解析

项目编号： No.31460369

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 食品科学、农学基础与作物学

项目作者： 白江平

作者单位： 甘肃农业大学

项目金额： 50万元

中文摘要： 蔗糖非酵解相关蛋白激酶家族2 (Sucrose Non-Fermenting Related Kinases，SnRK2) 是一类植物中高度保守的蛋白激酶，也是植物响应干旱盐碱等导致的渗透性胁迫的主要调控原件，但关于其在重要作物中的抗逆机理研究还很薄弱。本项目以马铃薯SnRK2类激酶为研究对象，利用TALEN (transcription activator-like effector nucleases) 基因组定向修饰与花粉原生质体转化相结合的技术手段构建SnRK2s敲除植株；利用In-Gel Kinase分析SnRK2s的体外激酶活性并确定非ABA依赖型SnRK2；利用荧光蛋白标记，实时定量RT-PCR，RNA-Seq等技术手段分析干旱胁迫下非ABA依赖型SnRK2激酶的组织特异性表达与遗传调控途径。研究结果将为解析马铃薯干旱胁迫响应代谢网络以及马铃薯抗逆育种提供理论依据与技术基础。

中文关键词： 马铃薯；SnRK2；花粉原生质体转化；TALEN；干旱胁迫

英文摘要： Sucrose Non-Fermenting-1 Related Kinases 2（SnRK2）is a family of highly conserved protein kinases in plants. They play important roles in the development of abscisic acid-dependent plants, and they are involved in the stress responses of plants to osmotic stress caused by drought or saline. To reveal the functional importance of SnRK2s in potato , we propose to employ the methods of TALEN (transcription activator-like effector nucleases) and combined with the pollen protoplast transformation technique to construct the SnRK2s knock-out potato plant. The physiological and biochemical characteristics of knock-out potato will be evaluated to reveal the regulatory function of SnRK2s. To study the activities of the SnRK2s in response to ABA, the His-tagged SnRK2s protein will be purified and the In-Gel Kinase assay will be conducted to analyze the in vitro activities of SnRK2s. To further characterize the function of non-ABA dependent SnRk2 in response to the abiotic stresses such as drought, the green fluorescence protein (GFP) labeling, real-time quantitative PCR will be employed to analyze the subcellular distribution and the stress induced tissue specific transcriptional expression. Final, the RNA-Seq will be conducted to analyze the regulatory roles of non-ABA dependent SnRk2. The results of this study should reveal the functional importance of non-ABA dependent SnRK2 in potato and would make it possible to explore the potentiality of improving the stress tolerance of potato through regulating the expression and activities of SnRK2s.

英文关键词： potato;SnRK2;pollen protoplast transformation;TALEN;drough stress