巨尾桉组培快繁与转基因再生体系的优化

Optimization of Eucalyptus grandis×E. ophylla tissue culture and regeneration system on genetic transformation

以巨尾桉带腋芽的半木质化茎段为最初外植体,通过正交试验优化法,对巨尾桉组培快繁体系建立过程中各项培养基进行了研究,建立起良好的巨尾桉组织培养再生体系,为桉树遗传转化和良种保存提供材料和技术支持,同时对根癌农杆菌介导的叶盘法转化体系进行优化。结果表明:叶片分化芽率提高到26.7%,理论上最佳的转基因巨尾桉叶片直接分化培养基为WPM+6-BA 0.3 mg/L+NAA 0.15 mg/L,生根培养基中筛选转基因芽段的卡那霉素选择压为20 mg/L。

Efficient tissue culture regeneration system of Eucalyptus grandis xE. ophylla was established by orthogonal optimization method. The stem-segments with axillary buds from aseptic seedlings were used as exp[ants to reproduce multiple buds. The system of agrobacterium-mediated transformation of Eucalyptus was optimized and the differentiation rate increased to 26.7%. Theoretically, the most suitable medium for direct differentiation of adventitious buds was WPM＋6-BA 0.3 mg/L＋NAA 0.15 mg/L and the Km resistance level of the transgenic plants was 20 mg/L.