中文摘要：

建立了根癌农杆菌介导的红豆杉细胞遗传转化体系，为红豆杉细胞的遗传改良打下基础。最优转化条件是：菌液光密度A600=0．6，侵染时间25min，共培养最适温度21℃，共培养时间48h；最适头孢霉素（Cephamycin，Cef）抑菌浓度为300mg／L；最优的除菌及筛选方式为：用含100mg／L头孢霉素的无菌水冲洗共培养后的细胞10s，然后将细胞转至舍300mg／L头孢霉素的抑菌培养基上，两周后转至含150mg／L卡那霉素（Kanamycin，Kan）或8mg／L潮霉素（Hygromycin，Hyg）的筛选培养基上筛选。采用小愈伤团法筛选转基因纯系，GUS染色显示，筛选后转化细胞株的阳性细胞占80％以上。

英文摘要：

In order to facilitate the genetic modification of Taxus cells, the genetic transformation system of Tax- us media cell mediated by Agrobacterium tumefaciens EHA 105 strain was established in the current work. The optimal bacterial concentration, infection time, temperature and co-culture time were A600 =0.6, 25 min, 21℃ and 48 h, respectively. The optimal cephalosporins chloramphenicol （Cef） bacteriostatic concentration was 300 mg/L. The most optimum methods for bacteria removing and Taxus cells screening were as follows： the co-cul- ture cells were washed for 10 s with distilled water containing 100 mg/L Cef, and then transferred to medium containing 300 mg/L Cef, and finally transferred to medium containing 150 mg/L kanamycin （Kan） or 8 mg/L hygromycin （Hyg） for further selection. A method of small callus culture was used to select the trans- genic pure cells. The results of GUS dyeing showed the ratio of the positive transformed cells in selected cell lines was above 80%.