抗草甘膦分基因cp4.docx

Expression Expression Vector Construction and Transformation of Glyphosate Resistance Gene CP4-EPSPS in Sweet Potato Abstract Sweet potato[Ipomoea batatas(L．)Lam】is one of important food crops and industrial crops．Its production is sufferring a lot from weeds damage，each year it can be cut down 5％一1 5％by weeds．The glyphosate resistant varieties was contained with the gene CP4-EPSPS transformed into sweet potato． In this study,the plant expression vector pCAMBIAl301--CP4··EPSPS was successfully constructed by connecting the cloning vector pMD 1 8·-T-CP4·—EPSPS with the plant expression vector pCAMBIAl301 containing CaMV35S promoter,NOS terminator,based on the amplification of gene CP4一EPSPS from a soybean variety with resistance to glyphosate．Then the stem section of sweet potato was infected with Agrobacterium tumefaciens harboring pCAMBIAl301·-CP4--EPSPS recombinant vector assisted by vacuum infiltration method．It ttLmed out that 5 of the 72 samples were identified to he positive by PCR verifivation，including 1 individual of Guangshu 87 and 4 individuals of Fushu 1 8．Glyphosate tolerance identification results showed that 3 individuals of Fushu 1 8 could tolerate glyphosate of 0．1 2％concentration．The above results was respected to facilitated breeding varieties、Ⅳitll resistance to glyphosate and commercial production of sweet potato and proved that it was feasible to transfer the stem section with vacuum infiltration assisted Agrobacterium tumefaciens mediated transformation method． Key Words：Sweet Potato，Glyphosate，Vector construction，Transformation II 万方数据 目 目 录 摘要 I Abstract．．．．．．． ．．．．． ．．． ． ．． ． ． ．．．．．．．．．．．．．．．．．．．． ． ． ．．．．．． ．．． ．．．．．．． ． ．．．．．．．．． ．．．．．．．II I前言 I 1．I甘薯基因工程育种 1 1．2甘薯遗传转化研究进展 2 1．2．1基因枪法 ．2 1．2．2电击法 ．2 1．2．3发根农杆菌介导法 ．2 1．2．4根癌农杆菌介导法 ．2 I．3转基因研究现状 3 1．3．1转基因作物 ．3 1．3．2抗除草剂转基因作物 ．5 1．3．3抗除草剂基因 ．5 1．4草甘膦及抗草甘膦转基因研究进展 6 1．4．1草甘膦概述 ．6 1．4．2草甘膦作用机理 ．7 1．4．3抗草甘膦作物的获得策略 ．．7 1．4．4抗草甘膦作物研究 ．9 1．5本研究的目的和意义 9 1．6本研究的创新点 9 2材料和方法 ．10 2．I材料 ．．10 2．1