| In this work, high frequency regeneration and transformation systems were established based on study of the factors affecting polar bulbs explants regeneration efficiency and gene transfer efficiency. IPT and NPR1 genes were introduced into Lilium through the improved protocols. The molecular detection and physiological analysis were also studied. The main research work and results were as follows:1. Construction of plant express vector pB35SNPR1 including NPR1 gene.NPR1 gene was injected into plasmid vector pGreen0229 to construct plant express vector pB35snpr1 (pGreen0229-35S-NPR1-GFP) selected noting by Kan. And then the new vector was further transferred into EHA105 by freeze-thaw method and used for transformation experiment of Lily.2. High frequence regeneration system of bulbs explants was established by orthogonal design. The results showed that the optimal time for interior bulbs was 9min, oppositely 15min for medium and exterior bulbs , and surviving rate reached 55%, 50%,45%, respectively, by 1‰ (W/V) Hg solution for explants killing agrobacteria after transformation. Meantime, the optimal regeneration medium was MS+6-BA2+NAA0.2mg/L for bulbs and MS+6-BA1+NAA0.2+2,4D0.1mg/L for leaves, and the regeneration rate reached 86.1%, 78.9%, respectively.3. Kan sensitive test indicated that the critical Kan sensitive concentration for inducing adventitious buds was 100mg/L, based on the regenerated leaves as materials for transformation system.Several crucial factors influencing the transformation efficiency were studied. It was found that pre-culture of leaves for 15-20 days before infection with A.tumefaciens (OD-0.5) for 15min, co-culture for 2 days after infection would be favorable for the transformation, the regeneration rate was 16% around; further investigated that 50μmol/L and 100μmol/L AS was supplemented into EHA105 and co-culture medium respectively would increase transformation rate.
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