苹果高效遗传转化受体系统的建立与优化

Establishment and Optimization of Acceptor System with High Frequency for Gene Transformation of Apple

以嘎拉（Gala）和长富2号（Nagafu No.2）组培苗离体叶片为外植体,进行了不同生长调节剂浓度、叶片成熟度、暗培养时间、叶片极性及抗生素筛选研究。结果表明,嘎拉叶片适宜的再生培养基为MS＋TDZ0.5 mg/L＋NAA 0.4 mg/L,叶片近轴面接触培养基;长富2号叶片适宜的培养基为MS＋TDZ 4.0 mg/L＋NAA 0.6 mg/L,叶片远轴面接触培养基。经过21 d的暗培养后,嘎拉和长富2号叶片再生频率分别可达100%和60%。抗生素筛选浓度为卡那霉素5 mg/L,头孢霉素200-300 mg/L。

Experiments of the concentration of growth hormone, maturity, dark treatment, leaf polarity and the concentration of the antibiotics were conducted with leaves of Gala and Nagafu No. 2 as in vitro explants. The results showed the Gala leaves were able to form buds on the MS medium containing 0.5 mg/LTDZ and 0.4 mg/LNAA with abaxial surface,adaixial surface of Nagafu No. 2 leaves touching MS medium with 4.0 mg/L TDZ and 0.6 mg/L NAA. After about 21d dark incubation treatment ,the generation efficiency of Gala and NagafuNo. 2 leaves reached 100% and 60%,respecticely. The concentration of Km and Cef were 5 mg/L and 200-300 mg/L.