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为探索DREB2ACA基因(constitutive activate DREB2A,组成型激活DREB2A基因)在植物抗盐反应中的功能及其在植物抗逆基因工程中的运用,用SOE-PCR(overlapping extension-PCR,折叠延伸PCR)方法克隆了DREB2ACA,并将其置于盐诱导的rd29A启动子控制下,构建了植物表达载体,并通过农杆菌介导的方法将目的基因转入烟草。Southern blot证明了目的基因成功整合到烟草基因组中。RT-PCR表明,目的基因受盐胁迫诱导表达。在盐处理条件下,转基因烟草比对照具有更好的生长状态,含有较高的光和色素,具有较高的光合速率及较低的MDA含量。说明DREB2ACA的表达提高了植物抗逆能力,该基因在抗盐领域具有广阔的应用前景。
In order to explore the function of DREB2ACA gene (constitutive activate DREB2A, constitutively activated DREB2A gene) in the plant salt-tolerance reaction and its application in plant stress-tolerant genetic engineering, we used SOE-PCR (overlap extension-PCR) DREB2ACA was cloned and placed under salt-induced rd29A promoter. Plant expression vector was constructed and transformed into tobacco by Agrobacterium-mediated transformation. Southern blot showed that the target gene was successfully integrated into tobacco genome. RT-PCR showed that the target gene was induced by salt stress. Under the salt treatment conditions, the transgenic tobacco had better growth status than the control, with higher light and pigment, higher photosynthetic rate and lower MDA content. This indicated that the expression of DREB2ACA enhanced the plant’s resistance to stress and the gene has broad application prospects in the field of salt resistance.