| 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1 cm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 mg/L分别与吲哚乙酸 0.05 ,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+BA 1.0 mg/L+IAA 0.2 mg/L为最佳生芽培养基 ,MS添加吲哚乙酸0.0,0.05,0.1和 0.2 mg/L进行生根比较 ,确定MS+IAA 0.05 mg/L为最佳生根培养基。卡那霉素(Kan)临界浓度确定为25 mg/L。转化培养过程为 :外植体于生芽培养基26 ℃ ,2 600 lx光下预培养24 h。农杆菌LBA4404过夜培养 ,用MS培养液稀释10～20倍 ,侵染外植体5 min,28 ℃黑暗共培养48 h。然后在MS+BA 1.0 mg/ L+IAA 0.2 mg/L+Kan 25 mg/L+Cef 200 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+IAA 0.05 mg/ L+Kan 25 mg/L+Cef 100 mg/L培养基上生根。 |
| 关键词: 番茄 组织培养 遗传转化 |
| DOI： |
| 分类号: |
| 基金项目:西北农林科技大学科研专项经费资助项目 |
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| Tomato plant regeneration in genetic transformation |
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| Abstract: |
| When tomato ZF seedlings grew 5-7 days after germination,cotyledons were cut into 0.5 cm×0.5 cm discs and hypocotyls into 1cm 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 2cm 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. |
| Key words: Tomato tissue culture genetic transformation |
