Study On Transformation Of BADH Gene In Sophora Japonica And Sophora Japonica Var Golden

There is a large of coastal and inland saline, so cultivating and alkali resistant germplasm selection breeding in the forest is an important goal. Sophora japonica and Sophora japonica var golden are of legume sophoral deciduous trees, belongs to temperate tree species, which is an excellent multi-purpose species and holds ecological and economic value. In this study in order to provide theoretical foundation for new germplasm further tests, some physiological and biological changes of them under salt stresses were tested, meanwhile made using of the Agrobacterium-mediated transformation of BADH gene in Sophora japonica to enhance its anti-reversibility, at the same time, BADH gene were transformed into Sophora japonica var golden to acquire the new species with the anti-salt. The results are as following:1. The study cultured the seedlings of transgenic Sophora japonica with rd29A gene and its rd29A-- recipient parents in the specialized nutrient solutions with NaCl contents, and then some physiological and biological changes of them were tested separately after different days. The result showed that the content of soluble sugar, free proline, malonaldehyde and electric conductivity were increased while the content of chlorophyll was decreased under NaCl treatments with the time. Under the salt stresses, the seedlings of the transgenic Sophora japonica showed higher osmosis- regulating contents, higher membrane stability and higher chlorophyll content compared with the non-transgenetic plants, so that they have stronger salt resistance.2. The study making using of the Agrobacterium-mediated transformation turns the BADH gene into Sophora japonica with rd29A gene. The results enunciation: the D600 of Agrobacterium tumefaciens was suitable for 0.3-0.5, infection time for 15 min, the callus infected by flat training method were cultured in the nutrient solutions without Kan for two weeks, then in the nutrient solutions with Kan for slecting. Whether a short time screening or long screening, soaking wound by AS solution is the most effective to improve budded rate. Through the above conditions, the transgenic plants with rd29A and BADH were obtained and conformed by PCR.3. In this study, taking leaves of Sophora japonica var golden as the explants, the optimum medium for the induction of adventitious bud differentiation was selected: MS + 6-BA 4 mg/L + IBA 0.5 mg/L. BADH gene was introduced into Sophora japonica var golden by Agrobacterium-mediated genetic transformation. After cocultivation of leaves of Sophora japonica var golden with Agrobacterium tumefaciens harbored BADH gene, kanamycin resistance shoots were transferred to selective MS medium containing 4 mg/L 6-BA, 0.5 mg/L IBA, 400 mg/L Cef and 100 mg/L Kan cefotaxime. PCR confirmed that BADH was successfully transfered into Sophora japonica var golden.