Establishment Of Regeneration System Of Lilium And Transformation With DREB2A Gene

In this paper,'Siberia'were used as material. The regeneration system of lilium was established. The scales were cultured to gain weight and their scale leaves were used to induce adventitious shoots. In the genetic transformation the scale leaves of lily were used as explants The stress promoter (DREB2A gene) was transferred into lilium mediated by Agrobacterium. The genetic transformation system of lily was established, and the positive plants detected by PCR analysis were obtained. The main results were as follows:1. Established the regeneration adventitious buds system though bulbs: The slice of Lilium bulbs were used as explants. MS+6-BA 2.0 mg/L+NAA 0.5 mg/L was advantageous to shoot induction, the rate of shoot induction was 85.0%; MS+6-BA 0.7 mg/L+NAA 0.2 mg/L was advantageous to proliferation; 1/2MS+NAA 0.2 mg/L was advantageous to rooting, the rate of root induction is 100.0%.2. Established the bulb gain system: MS+6-BA 0.2 mg/L+NAA 0.5 mg/L and sucrose concentration 7% was advantageous to bulb gain, proliferation rate was 291.7%. Established the regeneration adventitious buds system from scale leaf: the optimal system was: MS+6-BA 0.5 mg/L+2, 4-D 1.0 mg/L, which the regeneration rate was 92.5%; MS+6-BA 1.0 mg/L+NAA 0.5 mg/L was suitable to petiolule regeneration, and the regeneration rate was 83.3%.3. Established the genetic transformation system: Taking the scale leaf of Lilies as transformation receptor, pre-culture for 2 d; YEB as bacterial liquid, and liquid MS medium as the bacterial liquid of resuspension, infecting for 15 min with OD600=0.6 Agrobacterium tumefaciens solution, co-cultivation for 3 d, the concentration of Kan was 125 mg/L and the concentration of Cef was 400 mg/L in the process of sterilization, until the plantlet formed. Medium of co-cultivation supplemented with 50 mmol/L acetosyringone and subtracted NH4NO3 could improve the transformation rate.4. PCR detection of the transformed plant: Twenty seven resistant plants were obtained. Rate of resistance plant was 15.2%. Three of them were PCR positive plants, positive percentage was 11.1% .