钙调磷酸酶B类蛋白GmCBL1在大豆低温胁迫应答中的作用机制

项目名称： 钙调磷酸酶B类蛋白GmCBL1在大豆低温胁迫应答中的作用机制

项目编号： No.31201233

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 食品科学、农学基础与作物学

项目作者： 王俊峰

作者单位： 山东省农业科学院

项目金额： 20万元

中文摘要： 低温寒害是制约大豆栽培及生产的主要原因之一。钙调磷酸酶B类蛋白CBLs在植物钙离子信号传导途径中起重要调节作用，CBL1可能对低温胁迫应答基因的表达起负调控作用。CBL1的研究主要集中在拟南芥，而大豆GmCBL1的研究却很少。本研究选用本实验室收集筛选出的抗盐、低温敏感型大豆种质DAS7，建立了盐、低温胁迫及对照的基因表达谱，比较了它们的基因表达差异，筛选出对低温敏感的候选基因GmCBL1，拟研究其表达特异性、钙离子结合能力、亚细胞定位、非生物胁迫（盐、低温、干旱）及激素应答反应。将35S::GmCBL1分别转入拟南芥野生型及其同源基因缺失突变型，分析低温、激素处理下转基因拟南芥的生长及生理变化、低温胁迫应答相关基因表达变化，初步确定该基因功能；通过构建大豆酵母双杂交cDNA文库，筛选与GmCBL1互作的蛋白。初步阐述GmCBL1在低温胁迫应答中的作用机制，为耐寒大豆的分子育种奠定基础。

中文关键词： 钙调磷酸酶B类蛋白；CBL互作蛋白激酶；低温；钙离子；大豆

英文摘要： Low-temperature damage is one of the main constraints in soybean(Glycine max) cultivation and production. Calcineurin B-like proteins were crucially involved in relaying plant responses to calcium signal transduction pathway and in regulating ion fluxes. Researches had shown that negative regulation of CBL1 might be involed in the signal transduction pathway of cold response. CBL1 researches were mainly concentrated in dicotyledonous model plant Arabidopsis thaliana,while few researches in soybean CBL1.Soybean germplasm DAS7,salt-resistant but cold-sensitive, was selected in this research. Then DAS7 gene expression profiles were established under salt and low temperature stresses against the control by RNA-Seq. According to the obtained information, low-temperature-sensitive GmCBL1 was prepared to be the candidate gene for cloning from soybean. This research would be focused on expression and subcellular localization of GmCBL1,protein binding capacity with calcium ions, plant responses to abiotic (salt, low temperature, drought) and hormonal stresses. After 35S:: GmCBL1vector was constructed, it was transferred into wild type A. thaliana and AtCBL1 gene deletion mutant to validate the function of GmCBL1. Under low temperature and hormones treatments, it was analysed not only the changes of A. thaliana growth an

英文关键词： Calcineurin B-like protein；CBL-interacting protein kinases；Low temperature；Calcium ion；Glycine max