鲨烯合酶RNAi载体构建及对水稻的遗传转化研究  

Construction of RNAi Vector for Squalene Synthase Gene and Rice Genetic Transformation

作　　者：彭梅芳[1] 甘凤[1] 潘春梅[1] 陈克贵[1] 

机构地区：[1]四川省农业科学院生物技术核技术研究所,四川成都610061

出　　处：《西南农业学报》2015年第4期1413-1418,共6页Southwest China Journal of Agricultural Sciences

基　　金：四川省财政育种工程青年基金(2012QNJJ-002);四川省农业科学院生物技核技术研究所青年基金(SHS2012005)

摘　　要：鲨烯合酶(Squalene synthase,SQS)是ABA生物合成途径上游支路的一个关键酶。本研究克隆了406 bp的SQS基因片段,将其分别正向和反向插入干扰载体p YLRNAi.5中,经过酶切鉴定证实成功构建了干扰载体p YLRNAi-SQS,通过农杆菌介导法转入水稻品种Kasalath中,PCR初步鉴定获得了20株转基因阳性植株,进一步通过半定量RT-PCR分析,阳性转基因材料与野生型Kasalath相比,Os SQS3都有不同程度的下调,而Os SQS7变化不明显。证明转入的SQS基因只对Os SQS3起干涉效果,而对Os SQS7无作用,推测水稻中两个SQS基因可能存在不同的作用机制。本研究为进一步研究水稻中Os SQS3和Os SQS7各自的生物学功能奠定了基础,同时也为研究SQS调控ABA的生物合成提供了研究材料。Squalene synthase（ SQS） is a key branch point enzyme in ABA biosynthesis pathway. A 406 bp gene specific fragment of maize SQS was inserted into the p YLRNAi. 5 vector in forward direction and then in reverse direction,resulted in an RNAi vector,p YLRNAi-SQS.The recombined plasmid p YLRNAi-SQS was introduced into Agrobacterium tumefaciens strain LBA4404 to generate engineered strain. Rice Kasalath was genetically transformed with LBA4404 harboring p YLRNAi-SQS. Twenty transgenic plants with select marker gene,Hygr,were confirmed by PCR analysis among the regenerated plants. Semi-quantitative RT-PCR was carried out to investigate the expression of SQS genes in the transgenic rice plants,and results showed that the expression of Os SQS3 was down-regulated in several lines,but Os SQS7 was not in the same transgenic plants. All these lay a solid foundation for further research on biological function of Os SQS3 and Os SQS7,particularly in ABA biosynthesis in rice.

关 键 词：鲨烯合酶 RNAI 载体构建 遗传转化 水稻 

分 类 号：S511[农业科学—作物学]