颠茄高频再生体系的建立及卡那霉素抗性筛选

摘要

[目的]建立颠茄高频再生体系及筛选卡那霉素(Kan)抗性.[方法]以颠茄叶片及腋芽为外植体,研究了培养基中6-BA和NAA不同配比对其不定芽分化的影响以及叶片不定芽对Kan的敏感性.[结果]MS+4.5mg/L 6-BA+0.2mg/L NAA为叶片不定芽分化的最佳培养基,不定芽分化率达100%,在1cm×lcm叶块上的不定芽分化数平均达5.85;MS+3.0mg/L6-BA+0.1mg/L NAA为腋芽不定芽分化的最适培养基,不定芽分化率达100%,每个腋芽不定芽分化平均数为4～8个;400.0mg/L Kan为颠茄叶片遗传转化的最佳筛选浓度.[结论]为颠茄无菌苗的快速繁殖以及基于叶盘法的遗传转化奠定了基础.%[ Objective] The research aimed to establish the high frequency regeneration system of Atropa belladonna L. and screen the kanamycin (Kan) resistance. [ Method ] The leave and axillary buds of Atropa belladonna L. were the explants. The influence of different ratio of 6-BA and NAA in the medium on the adventitious bud differentiation and the sensibility of leaf adventitious bud on kan were studied. [ Result] MS + 2.5 mg/L 6-BA + 0. 2 mg/L NAA was the optimum medium of leaf adventitious bud differentiation, and the differentiation ratio of adventitious bud reached 100％. The number of adventitious bud differentiation on 1 cm× 1 cm leaf block averagely reached 5.85. MS +3.0 mg/ L 6-BA +0. 1 mg/L NAA was the optimum medium of axillary bud adventitious bud differentiation, and the differentiation ratio of adventitious bud reached 100％. The average number of adventitious bud differentiation in every axillary bud was during 4 - 8. The optimum kanamycin screening concentration of Atropa belladonna L. leaf genetic transformation was 400.0 mg/L. [ Conclusion] The research laid the foundation for the intermediate propagation of Atropa belladonna L. aseptic seedling and the genetic transformation based on the leaf disc cocultivation.