Research On Transformation Of Flower Color Gene Delila In Muscari Botryoides Mill.

Grape hyacinthus, also named grape lily, is a perennial herbaceous flower. It blossoms out very early and has a long florescence. They are very good garden flowers, and are usually cultured under tree covering the ground. They are also suitable for cutting flowers and pot culturing. The traditional breeding method need too much time, which not only low efficiency but single variety of color. Now cultured Grape hyacinthus are majority of hybrid variety of purple and red. They cannot appease people in need of many kinds of flower colors of Grape hyacinthus far and away. Breeding efficiency can be improved and new variety of aimed character can be acquired by transferring of exogenous genes. This study transferred Delila gene that control formatting red flower into somatic cell of grape hyacinthus"Album"by Agrobacterium tumefaciens in order to change the white flower into red flower.On the basis of the Delila expression vectors built, the system of callus and recipient was built in grape hyacinth"Album". Then transferring Delila gene to grape hyacinthus by Agrobacterium tumefaciens-mediated. For the purpose of achieving transgenic grape hyacinthus of Delila gene and provide some beneficial foundation in grape hyacinthus in the future .The results were as following:1.A regeneration system of leaf and bulb was initiated from Muscari botryoides"Album"Leaves and bulbs of Muscari botryoides"Album"were chosen as explants. Different type and concentration of hormone were studied in order to induce callus and adventitious bud. The results showed that the leaf is more appropriate explants for tissue culture of grape hyacinthus through comparing the bulb. The leaf and bulb were cultivated in No.1-10 mediums and they could grow callus in all treatment, but induced results were different. The optimum medium for inducing callus form leaves was MS+2,4-D 0.6 mg/L+6-BA 0.1mg/L, the induction frequency may reach 100%;The combination of MS+2,4-D 3.0 mg/L+6-BA 0.3 mg/L was the appropriate medium for bulbs forming callus with the induction rate 91.2%. The best medium for inducing adventitious buds from callus was MS+6-BA 2.0mg/L+NAA 0.3 mg/L, the germination rate was 97.4%;The medium of 1/2 MS+IBA 1.0 mg/L+1g/L AC was suitable for rooting with strong roots.2 A receptor system of callus was initiated from Muscari botryoides"Album" A recipient system of callus was initiated from Muscari botryoides"Album"with the regeneration leaf as explants. The combination of MS+2,4-D 0.6 mg/L+6-BA 0.1mg/L+sucrose 30 g/L was the optimum medium for the induction of calluses; the medium of MS+6-BA 2.0mg/L+NAA 0.3 mg/L+ sucrose 30 g/L was the best medium for regeneration adventitious buds from callus ,and the sensibility of callus to kanamycin was 60 mg/L. The carbenicillin used for eliminating Agrobacterium and the optimum concentration was 300 mg/L.3.An Agrobacterium tumefaciens-mediated transformation protocol was established for Muscari botryoides"Album"An Agrobacterium tumefaciens-mediated transformation protocol was established for Muscari botryoides"Album"by using the recipient system of callus as transformation recipient in grape hyacinth.4.Plantlet of grape hyacinthus containing Delila gene was obtained.According to PCR test, one positive plantlet was verified in 64 kanamycin-resistent buds from callus by Agrobacterium EHA105 and the frequency of transformation was 1.56%.