摘要
本研究对来自苏云金芽胞杆菌BT8的Cry1A(h)基因根据玉米密码子偏好性进行了优化。采用重叠PCR方法获得了优化的Cry1Ah基因与抗除草剂基因2m G2-epsps融合片段,并将其构建到植物表达载体p BI121上,构建p BI121-EPSPS-cryl Ah双元载体。将其转化根癌农杆菌菌株EHA105,获得基因工程菌。结果表明,玉米生产上使用的骨干优良自交系18599、综31等的幼胚作为转基因受体材料,以工程菌将抗虫抗除草剂双价基因转入上述玉米幼胚,经共培养、恢复、筛选、分化、生根、壮苗移栽等培养,苗期除草剂草甘膦涂抹,免疫试纸条检测、抗虫接种鉴定等初筛,再经PCR验证,获得了4株双抗转基因材料。此结果将为抗虫抗除草剂双抗转基因玉米新品种选育提供较好的亲本材料。
In this study, CrylA （h） gene from bacillus thuringiensis （BT8） was optimized based on the cedon bias of maize, the coalesce segment between CrylAh gene and the herbicide-tolerant gene（2mG2-epsps） were obtained by overlapping PCR technology. To construct the binary vector, the coalesce segment was combined with the plant expression vector pBI121, and the binary vector was transformed into the Agrobacter/um tumefaciens EHA105 as the genetically modified bacteria. The result showed that the embryo from superior inbred lines（ 18599 and 231 ） as the transgenic acceptors, which was integrated firm insect and herbicide resistance genes, was transformed into the embryo of maize. Through co-cultivation, renewing, selecting, differentiation, roofing, seedling and transplanting to cultivate the embryo, smearing glyphosate, the immune strip test and insect-resistant vaccination identification were applied to initial screening test during seedling period, finally, four double resistant materials were obtained by PCR analysis. The double resistant materials could be used as the excellent parents for transgenic maize breeding with the insect and herbicide resistance genes.
出处
《西南农业学报》
CSCD
北大核心
2015年第2期475-479,共5页
Southwest China Journal of Agricultural Sciences
基金
转基因生物新品种培育重大专项(2014ZX0800301B)
四川省农科院财政基因工程项目(2011JYGC01-002)
四川省农科院高新技术研究应用专项(2013GXJS-001)
关键词
玉米幼胚
遗传转化
抗虫基因
抗除草剂基因
Maize callus
Genetic transformation
Insect resistance sene
Herbicide resistance gene
