陆地棉GhVP1基因干扰载体的构建及拟南芥的遗传转化

Construction of the Interference Vector of GhVP1 Gene in Gossypium hirsutum L. and Arabidopsis Genetic Transformation

为进一步探索陆地棉GhVP1基因的功能,本研究以本实验室克隆得到的陆地棉GhVP1基因为模板,经Blast比对,选择长度为347 bp片段作为干涉载体正义、反义序列。利用In-fusion方法构建干涉载体,挑选阳性克隆测序验证,成功构建pBI121-PH::VP1干涉载体。利用农杆菌介导法转化拟南芥,并进行PCR检测,成功获得9株转基因苗,并对获得的T3代转基因拟南芥进行耐盐性分析。结果显示,盐胁迫下,转干涉载体拟南芥的发芽率降低,根长变短,整个生育期的生长受到抑制。研究表明VP基因与植物的耐盐性相关,为进一步研究GhVP1基因及其作用机理提供了参考依据。

In order to explore the function of GhVP1 gene in upland cotton further, in this study the upland cotton GhVP1 gene cloned by our lab was used as a template, after the Blast comparison, a fragment of 347 bp was selected as sense and antisense sequence. The interference vector was constructed with In-fusion method and positive cloning was selected to sequence, and finally p BI121-PH：：VP1 interference vector was built successfully.Arabidopsis thaliana were transformed by Agrobacterium tumefaciens-mediated, via the PCR testing, finally, 9transgenic seedlings were obtained. And then the salt resistance analysis of the T3 generation of transgenic Arabidopsis thaliana was analyzed. The results showed that under salt stress, the germination rate of transgenic seedlings reduced, the root length of transgenic seedlings shorten and the whole growth period of transgenic seedlings was restrained. The study showed that the VP gene was associated with the salt resistance of plants,which provided reference basis for the further study of GhVP1 gene and its function mechanism.