Stem Lump Mustard And Efficient Regeneration System And Genetic Transformation System Of Research

Medicinal plant tissue culture is not only the genetic engineering application in plant resources exploitation and utilization of important technical foundation, but also play an important role in medicinal plant resources protection and sustainable utilization. This paper mainly studies of the optimization of the tissue culture, the cloning and analysis of flowering gene cry2, and the genetic transformation system’s set up in Brassica juncea var. tumida Tsen et Lee which have widely used and medicinal value.The optimization of the tissue culture:Using Stem tumor mustard seeds as raw material, sterile tissue method,1/2MS culture training, the six days seeding age cotyledons were used as the explants, the effect of different hormone of6-BA、NAA、2.4-D reflect the rate on explant differentiation and callus into seedlings were studied with orthogonal design. The results showed that the best culture medium for the growth of stem lunp mustard cotyledon is MS with6-BA3mg/L, NAA0.10mg/L,2.4-D Omg/L. The results also indicated that1/2MS supplied with0.10mg/L NAA is good for seedling cultivation strike root. Regeneration frequency of optimization culture medium has greatly raised.The cloning and analysis of cry2gene:Cryptochrome have important impacts on plant flowering. The cryptochrome control seedling de-etiolation; mediated hypocotyl elongation; control of nutrition and anthocyanin accumulation; and the maintenance of plant endogenous rhythms in plants. The research devise cry2gene expand primer compare with relatives plant and construct of over-expression vector and RNAi vector based on cloning cryptochrome2gene from stem lunp mustard. Then successfully transformed into agrobacterium tumefaciens strain GV3101to form engineering strain.Genetic transformation system is established: Research different conditions of preculture, coculture, bacteria liquid concentration, dip time and Kan concentration reflect the rate on conversion using orthogonal design methods. The results showed that the best condition of genetic transformation is preculture2d, coculture4d, bacteria liquid concentration (OD600) is0.5, dip time is10min and Kan concentration is30mg/L.Establish the optimization of genetic transformation system set important fundation for the reserach of Brassica juncea var. tumida Tsen et Lee and cruciferous plant’s genetic transformation and resource development and utilization.