不同启动子调控转GhCDPK1基因烟草的抗逆性研究

为探究不同启动子对陆地棉GhCDPK1基因抗逆功能的影响,该研究克隆了长度为824bp和1 524bp的2个拟南芥RD29A的启动子序列,分别构建了35S启动子和2个RD29A启动子驱动的GhCDPK1融合表达载体,并利用农杆菌介导法转化烟草,分析了其驱动的转GhCDPK1基因烟草,在逆境胁迫处理后的表型变化,叶绿素、丙二醛(MDA)和脯氨酸含量,过氧化物酶(POD)和超氧化物歧化酶(SOD)活性以及细胞膜透性的生理变化。结果显示:RD29A启动子驱动的转GhCDPK1基因烟草,比35S启动子驱动表现出更强的耐逆性,其叶绿素含量、脯氨酸含量以及POD、SOD活性都高于35S启动子,而MDA含量与细胞膜的通透性低于35S启动子,且1 524bp的RD29A2启动子片段驱动转GhCDPK1基因烟草的耐胁迫能力比824bp启动子片段更强。

Study on Stress Resistance of Transfer GhCDPK1 Gene Tobacco Regulated by Different Promoters

In order to investigate the effects of different promoters on the resistance function of GhCDPK1 in cotton, we cloned two promoter sequences of Arabidopsis thaliana RD29A with length of 824 bp and 1 524 bp, and constructed the GhCDPK1 fusion expression vector driven by 35S promoter and two RD29A promoters, respectively. These expression vectors were introduced into wild type tobacco by Agrobacterium mediated leaf disk transformation method. After the drought, salt and low temperature treatments, the tobacco was analyzed through the changes of phenotypic and the content of chlorophyll, peroxidase activity, superoxide dismutase activity, MDA content, proline content and the physiological of membrane permeability. The results indicated that RD29A promoter driven transgenic tobacco showed stronger resistance than the 35S promoter driven transgenic tobacco. And the chlorophyll content, proline content, POD and SOD activities were higher than those of 35S promoter driven transgenic tobacco. But MDA content and cell membrane permeability were lower than those of 35S promoter driven tobacco. The GhCDPK1 transgenic tobacco driven by 1 524 bp promoter of RD29A2, was more resistant to stress than transgenic tobacco driven by the RD29A1 promoter of 824 bp.