Gene Expression And Functional Analysis Of TaCIPK8 In Transgenic Tobacco

Abiotic stress,such as drought and salinity,adversely affects normal growth of plants.Plants have developed a series of regulatory mechanisms to respond to these stresses in long-term evolution.Calcineurin B-like protein?CBL?and its interacting protein kinase?CIPK?play important roles in response to abiotic stress in plants,but there are few reports about the function of CBLs and CIPKs in wheat.In this study,we first analyzed the expression pattern of TaCIPK8,then using transgenic approach to study the function of TaCIPK8 in response to salt stress.The main results are as follows:?1?qRT-PCR results showed that TaCIPK8 was expressed at different levels in 9organs and TaCIPK8 had higher expression in stem and leaves.The expression of TaCIPK8 was upregulated in wheat leaves after the treatment with 200 mM NaCl,10 mM H₂O₂ and 100?M ABA.?2?Yeast two-Hybrid Assay was used to analyze the interaction of TaCIPK8 and TaCBLs.Results showed that TaCIPK8 could only interact with TaCBL1 among 7TaCBLs detected.?3?The over-expression vector pBI121-TaCIPK8 was constructed and it was transformed into tobacco plants by Agrobacterium mediated transformation method.Transgenic plants were identified via PCR detection.?4?Phenotypic indice analysis of TaCIPK8-overexpressing tobacco plants showed that the germination rate and root lengths of transgenic lines were higher and longer respectively,in comparison to the control lines during salt stress conditions.This shows that the salt tolerance of tobacco plants was increased as a result of the overexpression of TaCIPK8 gene.?5?The over-expression vector pAHC25-TaCIPK8 was constructed and it was transformed into young wheat embryos by Gene gun-mediated transformation method.Transgenic plants were identified via PCR detection.Our results showed that TaCIPK8 had a positive role in response to high salt stress in transgenic tobacco.It provided useful reference for further study of the function of TaCIPK8 in wheat.