小麦CBL结合蛋白激酶基因TaCIPK16的克隆及特征分析  

Characterization of a CBL-interacting Protein Kinase Gene TaCIPK16 in Wheat

作　　者：MYO THWIN 刘芃[1] 马微[1] 薛庆贺 郭军[1] 康振生[1] 

机构地区：[1]西北农林科技大学植物保护学院,旱区作物逆境生物学国家重点实验室,陕西杨陵712100

出　　处：《西北植物学报》2016年第6期1073-1079,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家“973”计划(2013CB127700);国家自然科学基金(31371889);教育部新世纪优秀人才支持计划(NCET-12-0471);高等学校学科创新引智计划(B07049)

摘　　要：类钙调磷酸酶亚基B蛋白(calcineurin B-1ike protein,CBL)作为一类钙离子结合蛋白,通过与一类蛋白激酶(CBL-interacting protein kinase,ClPK)结合,从而在钙信号依赖的生理生化过程中发挥作用。该研究在条锈菌诱导的小麦叶片中克隆获得CIPK家族中1个基因TaCIPK16,并利用qRT-PCR技术、酵母双杂交技术及亚细胞定位技术分析了其功能特性。序列分析表明,TaCIPK16编码447个氨基酸,包含保守的激酶催化结构域及调控结构域,与水稻、拟南芥CIPK蛋白具有高度相似性。酵母双杂交分析验证显示,TaCIPK16与TaCBL4和TaCBL9存在强烈互作。定量分析表明,TaCIPK16受到条锈菌的诱导表达,在小麦与条锈菌互作过程中呈显著差异表达趋势。综上结果,TaCIPK16可能作为正调控因子参与了小麦对条锈菌的抗病防卫反应。The calcineurin B-like（CBL）-CBL-interacting protein kinase（CIPK）network plays a pivotal role in regulating the physiological as well as developmental processes in plants.In this study,we obtained a CIPK gene,TaCIPK16,from the wheat leaves infected by Puccinia striiformis f.sp.tritici（Pst）using in silico cloning and RT-PCR.The full-length cDNA of TaCIPK16 was 1 606 bp,which encoding 433 amino acids.Multi-sequence alignment showed that TaCIPK16 share high similarity with OsCIPK16 and AtCIPK16in rice and Arabidopsis,respectively.Analysis of the protein domain features indicated that TaCIPK16 contained conserved regulatory domain in C-terminal and protein kinases domain in N-terminal.Subcellular localization assays revealed that TaCIPK16 displayed a localization pattern similar to that of the GFP control,indicating a cytoplasmic and nuclear localization.Yeast two-hybrid assays showed that TaCIPK16 strongly interacts with TaCBL4 and TaCBL9,respectively.qRT-PCR assays indicated that TaCIPK16 was induced by Pst infection and differentially expressed during incompatible and compatible interactions between wheat and Pst.Our results suggest that TaCIPK16 has a positive role in regulatingwheat resistance against Pst.

关 键 词：小麦 CBL 蛋白激酶 TaCIPK16 条锈菌 

分 类 号：Q785[生物学—分子生物学] Q786