小麦CBL结合蛋白激酶基因TaCIPK16的克隆及特征分析

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。

Characterization of a CBL interacting Protein Kinase Gene TaCIPK16 in Wheat

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 AtCIPK16 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 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 regulating wheat resistance against Pst.