Study On The Gene DREB Expression And Transformed Of Alfalfa

Alfalfa(Medicago sativa) is a perennial legume crop wildly planted in the world. It is not only the important forage crop,but also has a good ecological benefit to ecological environment protection and prevents soil erosion. The growth of alfalfa often has severely affected by the low temperature,drought and high salt stress. Therefore, research on improvement of alfalfa stress resistance is of practical significance. In this research, the DREB gene was transferred to"Zhongmu NO.1"by Agrobacterium-mediated transformation. The main achievements of the research are following:(1) Regeneration tissue culture system establishment of"Zhongmu No.1".The embryogenic callus induction was SH+2,4-D(0.2mg/L)+6-BA(0.5 mg/L);the subculture media was MSO+NAA(0.5 mg/L)+6-BA(0.5 mg/L)+AgNO3(1.0 mg/L); the differential medium was MS, and the effective root inducing meida was 1/2MS.(2) PBI121-DREB gene plant expression vector. We obtain DREB gene plant expression vector.In this study the DREB gene was transfered into"Zhongmu No.1".(3) The transformation system mediated by Agrobacterium tumefaciens had been studied and optimized systemically.The transformation system is that:preculture 3days, OD6000.6agrobacterial density, 10minutes infection time and5days coculture. the transfer to media with 350mg/LCef and 40mg/LKan.