番茄ZF遗传转化再生体系的研究  

Tomato plant regeneration in genetic transformation

作　　者：尹明安[1] 郭立[2] 刘华伟[2] 崔鸿文[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨陵712100 [2]西北农林科技大学生命科学学院,陕西杨陵712100

出　　处：《西北农林科技大学学报（自然科学版）》2002年第5期27-30,共4页Journal of Northwest A&F University(Natural Science Edition)

基　　金：西北农林科技大学科研专项经费资助项目

摘　　要：番茄 ZF无菌苗出芽后 5～ 7d,子叶切成 0 .5 cm× 0 .5 cm小块 ,下胚轴切成 1cm的小段作为外植体 ,以 MS为基本培养基 ,玉米素 1.0 mg/ L,6 -苄基腺嘌呤 1.0 m g/ L 分别与吲哚乙酸 0 .0 5 ,0 .2 ,0 .5和 1.0 mg/ L配成 7个激素组合 ,经诱芽比较 ,确定 MS+BA1.0 m g/ L+IAA0 .2 mg/ L 为最佳生芽培养基 ,MS添加吲哚乙酸0 .0 ,0 .0 5 ,0 .1和 0 .2 mg/ L 进行生根比较 ,确定 MS+IAA0 .0 5 mg/ L 为最佳生根培养基。卡那霉素 (Kan)临界浓度确定为 2 5 mg/ L。转化培养过程为 :外植体于生芽培养基 2 6℃ ,2 6 0 0 lx光下预培养 2 4 h。农杆菌 L BA4 4 0 4过夜培养 ,用 MS培养液稀释 10～ 2 0倍 ,侵染外植体 5 m in,2 8℃黑暗共培养 4 8h。然后在 MS+BA1.0 mg/ L+IAA0 .2 m g/ L+Kan2 5 mg/ L+Cef2 0 0 mg/ L 筛选培养基上诱芽 ,每 14 d转接 1次。芽长至 2 cm时 ,切下转至 MS+IAA0 .0 5 mg/ L+Kan2 5 m g/ L+Cef10 0 m g/ L 培养基上生根。When tomato ZF seedlings grew 5-7 days after germination,cotyledons were cut into 0.5 cm× 0.5 cm discs and hypocotyls into 1 cm segments as explant.MS was used as basic medium,and Zeatin 1.0 mg/L and BA 1.0 mg/L were resperctively combined with IAA 0.05,0.2,0.5 and 1.0 mg/L as additions.It was determined that MS+BA 1.0 mg/L+IAA 0.2 mg/L was the best shooting medium.IAA 0.0 ,0.05,0.1 and 0.2 mg/L were respectively added into MS medium and rooting effects were tested.Results showed that the best rooting medium was MS+IAA 0.05 mg/L.Optimum concentration of Kan was 25 mg/L.Process of transformation culture was as followings:The discs and segments were pre cultured in shooting medium for 24 h (26 ℃,2 600 lx).Agrobacterium LBA4404 was cultured overnight in YEB and diluted 10-20 times with MS medium,in which explants were soaked for 5 minutes.Then the exptants were co cultured for 48 h (28 ℃,in the dark).After that,the explants were cultured in medium MS+BA 1.0 mg/L+IAA 0.2 mg/L+Kan 25 mg/L+Cef 200 mg/L for selection with transfer every two weeks.When shoots were 2 cm high,they were cut and transferred into the medium MS+IAA 0.05 mg/L+Kan 12.5 mg/L+Cef 100 mg/L for rooting.

关 键 词：番茄 ZF 遗传转化 再生体系 组织培养 

分 类 号：S641.2[农业科学—蔬菜学]