[Objective] The research aimed to establish the high frequency regeneration system of Atropa belladonna L. and screen the kanamycin （Kan） resistance. [Method] The leaves and axillary buds of Atropa belladonna L. were as the explants,the influences of different ratios of 6-BA and NAA in the medium on the adventitious bud differentiation and the sensibility of leaf on Kan were studied. [Result] MS＋4.5 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. 0 cm ×1. 0 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 screening concentration for genetic transformation of Atropa belladonna L. leaf was 400.0 mg/L of Kan. [Conclusion] The research laid the foundation for the rapid propagation of Atropa belladonna L. aseptic seedling and the genetic transformation based on the leaf disc cocultivation.
