农杆菌介导的大豆萌发种子转化技术  

The Technique for Agrobacterium-mediated Transformation via Germinating Seeds of Soybean

作　　者：刘志敏[1] 熊鹤雯[1] 谢皓[1] 秦玉涛[2] 赵嫣然[2] 郭蓓[2] 

机构地区：[1]北京农学院植物科学技术学院,北京102206 [2]北京农学院生物科学与工程学院,北京102206

出　　处：《生物技术通报》2013年第8期83-87,共5页Biotechnology Bulletin

基　　金：国家高技术研究发展计划"863"项目(2012AA101106)

摘　　要：采用农杆菌介导的萌发种子侵染法,开展大豆遗传转化技术研究。以萌发大豆的生长点、子叶节、下胚轴3处为外植体,将带有抗除草剂草甘膦基因EPSPS及草丁膦抗性筛选基因Bar的质粒借助农杆菌介导转入大豆中,通过共培养及盆栽种植获得转化体。经过对T0代和T1代植株进行双重PCR检测以及两种除草剂的抗性鉴定得到转基因阳性株。试验结果表明,通过农杆菌介导的萌发种子转化法已将抗两种除草剂基因成功的转入大豆中,其综合转化率约为0.37%。Germination seeds were used to research the genetic transformation technology of Agrobacterium-mediated in soybean.The growing point,cotyledon node and hypocotyl of germination seed were used as explants,and the plasmid that carried with the resistance to the herbicide glyphosate gene EPSPS and glufosinate resistance screening gene Bar was transferred to soybean by Agrobacterium-mediated.Transformants were obtained by co-culture and potted plant growing.After the double PCR detection as well as the two herbicide resistance identification of transgenic T0 and T1 plants,the positive strains were acquired.The experiment results showed that two genes resistance to herbicides have been successfully transferred to soybean via germinated seeds by Agrobacterium-mediated transformation method,with the transformation rate of approximately 0.37%.

关 键 词：农杆菌 萌发种子 大豆 遗传转化 

分 类 号：S565.1[农业科学—作物学]