农杆菌介导的木薯转基因研究初报  

Preliminary report on genetic transformation of cassava mediated by Agrobacterium tumefaciens

作　　者：覃艳[1] 何勇强[2] 农友业[1] 姜伯乐[2] 吴子恺[1] 

机构地区：[1]广西大学农学院,广西南宁530005 [2]广西大学生命科学与技术学院,广西南宁530005

出　　处：《广西农业生物科学》2008年第3期218-222,共5页Journal of Guangxi Agricultural and Biological Science

基　　金：广西科学研究与技术开发计划(桂科基0009017)

摘　　要：将米曲霉植酸酶基因phyA克隆于植物表达载体pBI121的CaMV35S启动子(P35S)下游,构建了含有P35S-phyA-TNOS表达盒的重组质粒pBI-phyA,并用电脉冲法将其导入大肠杆菌中。通过三亲接合将pBI-phyA质粒由大肠杆菌导入根癌农杆菌LBA4404菌株,获得带有phyA基因的根癌农杆菌工程菌株LBA4404/pBI-phyA。用LBA4404/pBI-phyA转化木薯外植体652个,在含100 mg/L卡那霉素和250 mg/L氨苄青霉素的MS分化培养基上诱导分化芽,获得24株抗卡那霉素转化苗。以转化苗和未转化植株的总DNA为模板,用phyA基因的同源序列为引物,进行聚合酶链式反应(PCR),结果表明9株转化苗的PCR结果为阳性,初步证明目的基因phyA已经转入木薯中。The Aspergillus oryzae phytase gene phyA was cloned into the site of downstream the CaMV35S promoter （P3ss） of the plant expression vector pBI121, generating a recombinant plasmid pBI-phyA containing the expression cassette P35s-phyA-TNOS. pBI-phyA was then introduced into the Escherichia coli cells by electroporation. The genetic engineering Agrobacterium tumefaciens strain LBA4404/pBI-phyA was constructed by introducing the plasmid pBI-phyA LBA4404 from E. coli/pBI-phyA via triparental conjugation. 652 explants of cassava （Manihot esculenta Crantz） were co-cultured with LBA4404/pBI-phyA. Regenerated shoots were selected out on MS medium supplemented with 100 mg/L kanamycin and 250 mg/L ampicilin. 24 putative transgenic plants were obtained. The total genomic DNAs of these regenerated plants and untransformed plants were detected with polymerase chain reaction （PCR） . The result indicated that 9 of these regenerated plants contained phyA gene.

关 键 词：植酸酶基因 根癌农杆菌 木薯 转基因 

分 类 号：S533[农业科学—作物学]