AhNCED1基因转化花生研究

Transformation AhNCED1 Gene in Peanut(Arachis hypogaea L.)

构建转化AhNCED1基因花生(Arachis hypogaea L.)过表达载体35S∷AhNCED1∷GUS,用OD600=0.8的LBA4404农杆菌液浸染汕油523,抗性芽诱导率达100%。PCR检测89株筛选苗,43株呈阳性,GUS检测阳性率为50%。转基因植株地上部分ABA含量增加;PEG胁迫10 h,转基因植株叶片AhNCED1蛋白表达增强,内源ABA水平积累,超氧化物水平降低。

9-cis-epoxycarotenoid dioxygenase（NCED） is the key enzyme of ABA biosynthesis in higher plant.The expression vector 35S∷AhNCED1∷GUS for peanut（Arachis hypogaea L.） was constructed.Then AhNCED1 gene was transformed into cotyledon of ＇Shanyou 523＇,which is sensitive to drought stress,by immersing into Agrobacterium tumefciens strain LBA4404 bacterial suspension solution.The induction rate of adventitious buds with kanamycin-resistant was 100%,and in which there were 48.3% and 50% positive plantlets by PCR amplification and GUS histochemical identification,respectively.The ABA contents in aboveground organs of transgenic plants were higher than that in the wild（non-transformation） plants.When the leaves of the transgenic AhNCED1 plants subjected to PEG stress for 10 h,the expressions of AhNCED1 protein and endogenous ABA content were increased,as well as superoxide level decreased.