Abstract: Lettuce cultivar Hongkong glass lettuce was used as the test material to establish a high frequency regeneration and rapid propagation system. Effects of different sterilization methods on disinfection of seed, the growth of seedling， and the relationship between different hormones and medium on the induction callus of cotyledon， differentiation and rooting were studied. The results were as follows: When the lettuce seeds were treated for 60 s with 70% ethanol， followed by 2% NaClO for 10 min， the germination rate reached 98%， and the sterile seedlings grew well on MS medium. 3-5 d cotyledons should be selected as explants， and the optimal condition of adventitious buds was MS medium combined with 01 mg·L-1 TDZ and 01 mg·L-1 NAA while the differentiation rate reached 833% and differentiation index was 77. The medium of MS+ 03 mg·L-16\|BA+01 mg·L-1 NAA was optimal for the following generation and proliferation culture. The combination of NAA and IAA was beneficial for rooting， and the optimal medium for rooting was 1/2MS medium with 005 mg·L-1 NAA and 05 mg·L-1 IAA.

Key words: lettuce, seed sterilization, regeneration system