超长链多不饱和脂肪酸在棉花中的异源合成  

Production of Very Long Chain Polyunsaturated Fatty Acids in Cotton

作　　者：刘江[1] 马燕斌[2] 孙全喜[1,3] 吴霞[2] 李雪滢[1] 孙美红[1] 李燕娥[2] 李新征[1] 亓宝秀[1] 

机构地区：[1]作物生物学国家重点实验室/山东农业大学生命科学学院,山东泰安271018 [2]山西省农业科学院棉花研究所,山西运城044000 [3]山东省花生研究所,山东青岛266100

出　　处：《作物学报》2014年第1期86-92,共7页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育科技重大专项(2009ZX08005-024B);国家自然科学基金项目(30970222);山东省现代农业产业技术体系建设专项资金资助

摘　　要：从球等鞭金藻、眼虫、高山被孢霉和拟南芥中分别克隆到Δ9链延长酶、Δ8去饱和酶、Δ5去饱和酶和Δ15去饱和酶基因,利用我们的多基因聚合方法,将这4个基因聚合到植物表达载体pCambia2300上,其中每个基因都含有独立的CaMV35S启动子和Tnos终止子。利用农杆菌介导法将该表达载体转入棉花,通过卡纳霉素和PCR筛选获得转基因阳性植株,提取转基因阳性植株叶片总脂肪酸,用气相色谱分析法检测到花生四烯酸(AA,20:4Δ5,8,11,14)和二十碳五烯酸(EPA,20:5Δ5,8,11,14,17)含量分别达1.0%和5.0%。表明通过基因代谢工程在棉花中异源合成EPA是可行的,为进一步在棉籽中生产VLCPUFAs奠定了基础。We have isolated four genes encoding a Δ9 elongase,a Δ8 desaturase,a Δ5 desaturase,and a Δ15 desaturase from Isochrysis galbana,Euglena gracilis,Mortierella alpina,and Arabidopsis thaliana respectively.Using a multigene transfer tech-nology that we developed,these genes were stacked together in the plant expression vector pCambia2300.Each gene contained its own CaMV35S promoter and Tnos terminator.This plant expression vector was then transferred into cotton by Agrobacte-rium-mediated transformation method.Transgenic cotton seedlings were first identified by screening them based on kanamycin-containing media and followed by PCR with gene-specific primers of the four transgenes.Finally,these transgenic plants were subjected to gas liquid chromatography analysis for their fatty acid composition and the results showed that the contents of arachidonic acid（ARA,20：4Δ5,8,11,14） and eicosapentaenoic acid（EPA,20：5Δ5,8,11,14,17） were 1.0% and 5.0% respectively in the leaves of the transgenic plants,indicating that the four genes were expressed in cotton.Therefore,our data clearly demonstrated the feasibility for the heterologous production of EPA in cotton and this will lay a foundation for the production of VLCPUFAs,including EPA and DHA in cotton seed through transgenic technology in the future.

关 键 词：去饱和酶 链延长酶 转基因 EPA 棉花 

分 类 号：S562[农业科学—作物学]