| Lilium.pumilum in Liliaceae is a kind of northeast wild flower in alpine region, which has high application value. Lilium.pumilum has scattered on the mountains under the shady woodland with strong adaptable resistance.It's not only one of the important lily parents but also excellent landscape plant material. At present, natural distribution of Lilium volume has dropped severely because of ecological environment destruction.Therefore, the preservation of wild germplasm has become more and more important.In this study, the high regeneration of Lilium. pumilum was achieved via organogenesis, callus indirect organogenesis by using bulb scales and the floral organs (including the petal, receptacle, filament, ovary, stigma) as explants through plant tissue culture technique; the optimal condition for Lilium.pumilum seed germination was found and sterile seeds were conducted polyploidy induced by colchicine. Polyploid plants were identified by multiple chromosome.The study play a fundamental role in propagate and polyploid breeding of Lilium.pumilum even more further molecular breeding. The primary research results as follows:1. Bulb scales of Lilium.pumilum were cultured on MS medium supplemented with different plant growth regulators for mass propagation. The results showed that the MS medium containing 0.5 mg/L NAA and 1.5 mg/L 6-BA was suitable for bud differentiation. The bud differentiation rate was 100% and adventtitious buds grew healthily.The proliferation of adventtitious buds on MS medium containing 0.2 mg/L NAA and 1.0 mg/L 6-BA was achieved 3.7.1/2MS medium supplemented with 0.5 mg/LIBA was applied for the root developed with 100% survival rate.There is position effect during induction of differentiation in the scales, outer> middle> inner; lower> middle> upper.2. The regeneration of Lilium.pumilum floral organs was achieved via callus indirect organogenesis. The results showed that MS medium supplemented with 0.5 mg/L NAA and 0.5mg/L 6-BA was best applied for receptacle callus induction; MS medium supplemented with 0.2 mg/L NAA and 1.0mg/L was best applied for ovary callus induction; MS medium supplemented with 0.5 mg/L NAA and 1.0mg/L 6-BA was best applied for stigma callus induction;MS medium supplemented with 2 mg/L 2,4-D and 2 mg/KT was best applied for petal callus induction; the best medium for filament callus induction was the same as the medium of petal; the best medium for adventitious bud differentiation was MS medium containing 0.5 mg/L NAA and 2.0 mg/L 6-BA. Callus induction were different from easy to difficult, the order was receptacle> filament> ovary> stigma> petal. MS medium containing 0.2 mg/L NAA and 2.0 mg/L 6-BA was the best choice for receptacle callus and adventtitious buds proliferation. IBA can significantly enhance the effectiveness of rooting with concentration of 0.5mg/L, up to 100% survival rate.3. The research showed that the optimum seed germination condition was MS medium with 25℃temperature. Adding sterile water artificially can effectively guarantee a high seed germination rate.4. Colchicine was the effective treatment to multiple the Lilium.pumilum seed with concentration of O.lmg/L during 24 hours, the mutation rate was up to 30%. The mutation plants had thicker leaves, increased type, sturdy plant, vigorous growth. Through stomata observation, tetraploid plants had increased stomatal but density decreased. Dealing with root tip, the tetraploid plants were successfully obtained with multiple chromosome number of 48.
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