Establishment Of Regeneration And SOD Gene Transformation System Of Lily (L.Longifloum) Leaf

These are beautiful flowers, Lilies are suitable for use in the garden ;a shrub border;potted plant.lily bulbs have rich nutrition,is a kind of important food and medicinal plant.Under nature conditions of growth and development, plants were invariably exposed to different environmental stress, which had profound effects on crop production. Many studies had examined that modification of Mn-SOD expression in transgenic plants can improve plant stress tolerance. In this study, we had introduced the Mn-SOD gene into lily to evealuate its effect on the persistence of environmental stress. The transformed lily derived from leaf explants would pave the way for its breeding via genetic engineering.The commercial Easter Lily cultivars "white elegance"in vitro leaves of seedlings were selected as experimental materials in this study, basal medium, 6-BAand NAA as factors, an idea medium which was MS+6-BA1.0~2.0mg/L+NAA0.2mg/L+sucrose30g/L for leaf regeneration in"white elegance" was sifted through test. And a system for in vitro high efficient regeneration was established after some factors affecting on leaf regeneration had been studied,example: Effect of leaf part on regeneration; Effect of leaf size on regeneration .For the multiplication and root development of the adventitious bud from leaf disc, hormone concentration and composition were also examined in this study.On the base of regeneration system from leaf explant of "white elegance", several factors that affected genetic transformation of the Easter Lily cultivars were examined, including selection of Kan concentrations, the suspension of strain EHA105, concentrations of Carb, and so on. Thus a simple and efficient genetic transformation system was developed. Selecting 5~8mm Leaf base of stronger leaves as explants, then cocultured on the differentiation medium with Agrobacterium tumefaciens EHA105 harboring a binary vector containing chimeric genes of Npt, Gus report gene and Mn-SOD gene driven by CaMV35S promoter. After 3 days cocultureation, these explants were transferred to the differentiation-resistant medium to select transgenic shoots. Some regenerated Kanamycin-resistant plants were obtained by multiplying and rooting. Through examination of GUS report gene, it was showed that the GUS report gene was showed.