GhCDPK4基因的克隆和功能分析

为了研究棉花中GhCDPK4基因在响应非生物胁迫中所起的作用，通过PCR的方法克隆GhCDPK4基因，利用基因重组技术，构建植物过表达载体，采用农杆菌介导的叶盘法转化模式植物烟草，分析干旱和盐胁迫处理对转基因烟草表型和生理生化指标的影响。本研究成功克隆了属于棉花CDPK家族的基因GhCDPK4，构建了植物过表达载体pCAMBIA2300-GhCDPK4。实时荧光定量PCR(qRT-PCR)检测发现转基因烟草中GhCDPK4基因高水平表达，并且转基因烟草相比于野生型烟草表现出较强的耐旱和耐盐性，其中SOD、POD和CAT活性显著升高，而相对电导率和MDA含量降低。研究结果表明GhCDPK4基因可正向参与应答干旱和盐胁迫。

Cloning and Functional Analysis of GhCDPK4 Gene

To study the role of GhCDPK4 gene in response to abiotic stressin cotton.GhCDPK4 gene was cloned by PCR,and the plant overexpression vector was constructed by gene recombination technology.The plant overexpression vector was transformed into tobacco by Agrobacterium-mediated leaf disk method.Finally,the effects of drought and salt stress on phenotype and physiological and biochemical indexes of transgenic tobacco were analyzed.In this study,the gene GhCDPK4 belonging to the CDPK family was successfully cloned from upland cotton,and the plant overexpression vector pCAMBIA2300-GhCDPK4 was constructed.High-level expression of GhCDPK4 gene in transgenic tobacco was detected by quantitative RT-PCR (qRT-PCR).After drought and salt stress treatment,the tolerance of transgenic tobacco to drought and salt stress increased,and the activities of SOD,POD and CAT increased significantly,while the relative conductivity and MDA content decreased.The results showed that GhCDPK4 gene was positively involved in response to drought and salt stress.