【作者】 王敬乔； 李根泽； 陈薇； 和江明； 董云松； 曾黎琼； 寸守铣；

【Author】 Jingqiao Wang;Genze Li;Wei Chen;Jiangming He;Yunshong Dong;Liqiong Zheng;Shouxian Cun Biotechnology Institute of Yunnan Agriculture Academy, Kunming, 650223

【机构】 云南省农科院生物技术研究所；

【摘要】 通过对转化材料的预培养处理、外植体的选择与制备、外植体与农杆菌的共培养等条件的优化组合,建立了农杆菌介导的甘蓝型油菜(Brassica napus)高效转基因平台:(1)高效率的外植体制备,每批次实验可获得2000～5000个外植体;(2)高频率的基因转化,外植体愈伤组织阶段基因转化率为68%～83%;(3)较高效率的植株再生,转基因植株再生频率17%～27%;(4)高效率的植株筛选,转化植株筛选效率高于85%;(5)高效率的工作程序,用液体培养基部份地取代固体培养基,大大地减少了基因转化的工作量,单人单批次转化实验可获得转化植株300～700株;单人每年可获得转基因油菜植株5000～10000株。利用这样的转基因平台可进行无选择基因转化,且再生植株中转基因植株的比例可达20%左右;因此,该平台技术可用于直接生产无选择标记转基因植株。更多还原

【Abstract】 This paper reported a high--efficient transgenic platform and a method of nonselective gene transformation for Brassica napus mediated by Agrobacteriumtumefaciens. By this platform, a trained person is able to finish transformation work oftwo to five thousand explants and obtain three to seven hundred transgenic plants in eachtest. So, five to ten thousand transgenic plants can be produced every year by one person.Furthermore, the nonselective transformation of Brassica napus was tested by this high-efficient transgenic platform and transgenic plants were obtained with the efficiency of20% or so. This result suggests that the nonselective gene transformation may become aroutine method for producing marker--free transgenic plants after the selectable markergene is cut from T--DNA, at least for Brassica napus.更多还原