BADH Gene Import And Physiological Identification Of Salt-tolerant Materials Of Alfalfa

In this research, Three alfalfa varieties salt endurance mutant and normal callus were used as plant materials. The salt tolerance were identified by quantitative analysis and comprehensive evaluation of physiological and biochemical indicators (MDA, PRO, SS, SP, SOD and others) after NaCl solution stress(0, 0.2, 0.4, 0.6, 0.8, 1.0%). The BADH gene was transferred to alfalfa by Agrobacterium mediated transformation in this paper. The main results were as following:1. Results showed that the content of the MDA, proline, soluble protein and soluble sugar content, as well as superoxide dismutase (SOD) activity of the salt resistant mutants were significantly (p<0.05) or highly significant (p <0.01) higher than comparison material.2. The content of PRO and soluble protein increased follow by the content of Nacl; the content of Malondialdehyde(MDA) decreased first and then increased ,; the content of soluble sugar and the activity of Superoxide dismutase(SOD) increased first and then decreased on the whole . The transition in the change curve appeared at 0.8% concentration of Nacl on the whole.3. The order of salt tolerance was Millennium > CW787> Victoria.4. Cultivated 7 days hypocotyls of Gan-nong No.4 and Millennium as explants and agrobacterium tumefaciens strain as mediator , the optimized genetic transformation system is as follows: 200 mg·L^-1 CEF could restrained growing of agrobacterium effectively on the whole .The selective pressure of Gan-nong No.4 was 50mg L^-1and the Millennium was 50mg L^-1.The test object was hypocotyl. OD 0.3～0.5 agrobacterial density and 10 minutes infection time. In a series of replicated experiment, 10 Kanamycin resistant of Gan-nong No.4 were obtained. The result of PCR analysis stated that the detected Kanamycin resistant plants transgenic BADH gene was one positive. It showed BADH gene get a good expression in transgenic plants.