小麦CBL结合蛋白激酶基因TaCIPK16的克隆及特征分析

摘要

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 Pucciniastriiformis f.sp.tritici (Pst)using insilico cloning and RT-PCR.The full-length cDNA of TaCIPK16 was 1 606 bp,which encoding 433 a-mino acids.Multi-sequence alignment showed that TaCIPK1 6 share high similarity with OsCIPK1 6 and AtCIPK1 6 in 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 TaCIPK1 6 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 TaCIPK1 6 was induced by Pst infection and differentially expressed during incompatible and compatible interactions between wheat and Pst.Our results suggest that TaCIPK1 6 has a positive role in regulating wheat resistance against P st .%类钙调磷酸酶亚基B蛋白(calcineurin B-1 ike 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,TaCIPK 16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,TaCIPK 16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。