Optimization Of High Efficient Regeneration System Of Lilium And Agrobacterium-mediated Genetic Transformation With LIDREB1 Gene

As a kind of plant with elegant and fragrance, Lliy is usually used for cut flower and pot culture in commercial cultivation. Due to the destruction by the human being, the resources of wild lily are more serious. So it is very important to establish high efficient regeneration system of lily. Lily still has some characteristics that need to be improved owing to the customer’s demand. There are many breeding methods,but due to the shortage of traditional breeding, it should be make breakthrough in molecular breeding. Some achievements are being developed in Agrobacterium-mediated genetic transformation to change its features, But some difficulties are still exist in genetic transformation of Lilium plants due to its complicated background of hereditary. This study took different lily as material to establish regeneration system, it also took the study in Agrobacterium-mediated genetic transformation with aim gene. The main results were summarized as follows: 1. Optimization of regeneration system of Lilium longiflorum: When we use the bulb transverse thin cell layer as explants, proper medium for regeneration of Lilium longiflorum is 1.5MS+1.0mg/L BA+0.2mg/L NAA+90g/L sucrose+7.5g/L agarose,and it goes the way of multiple shoot clumps; When we use the bulb transverse thin cell layer or leaves as explants, proper callus induction medium is MS+1.0mg/L NAA+0.2mg/L TDZ+60g/L sucrose+7.5g/L agarose, callus differentiation medium is MS without any hormone; When we use the bulb transverse thin cell layer as explants, proper embryonic callus induction medium is MS+2.0mg/L PIC+ 0.1mg/L TDZ+60g/L sucrose+7.5g/L agarose,and it goes the way of embryonic. 2. Optimization of regeneration system of Lilium sulphureum:When we use the bulb transverse thin cell layer as explants, proper medium for regeneration of Lilium sulphureum is 1.5MS+1.0mg/L BA+0.6mg/L NAA+90g/L sucrose+7.5g/L agarose; and it goes the way of multiple shoot clumps; When we use the bulb transverse thin cell layer as explants, proper callus induction medium is MS+1.0mg/L NAA+0.2mg/L TDZ+60g/L sucrose+7.5g/L agarose,While we use the leaves as explants,proper callus induction medium is MS+0.5mg/L NAA+0.4mg/L TDZ+60g/L sucrose+7.5g/L agarose.Callus differentiation medium is MS without any hormone. 3. Embryonic callus of Lilium longiflorum are used as transformation materials, the target gene is LlDREB1, 32 resistant plants were obtained after selection.We got 6 PCR positive plants from 32 resistant plants,which were trans formed with EHA105. The rate of PCR positive is 18.8%.