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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 further explore the function of GhVP1 gene in Gossypium hirsutum, GhVP1 was cloned by our lab and aligned by Blast. The 347 bp fragment was selected as the sense and antisense vector of interference vector. The interfering vector was constructed by In-fusion method and positive clones were selected and sequenced. The pBI121-PH :: VP1 interfering vector was successfully constructed. Arabidopsis thaliana was transformed by Agrobacterium tumefaciens method and PCR assay was carried out. Nine transgenic plants were successfully obtained, and the salt tolerance of T3 transgenic Arabidopsis was analyzed. The results showed that under salt stress, the Arabidopsis transgenic Arabidopsis thaliana had lower germination rate, shorter root length and inhibited growth during the whole growth period. Studies have shown that VP gene is related to plant salt tolerance, which provides a reference for further study of GhVP1 gene and its mechanism of action.