Agrobacterium-Mediated Transformation Of Potato With Salt Resistance Gene BADH

In this study, BADH gene was transferred into Favorite, one main local cultivars, with agrobacterium-mediated method.The explants were microtuber discs. The transgenic plants were tested by PCR and Southern blotting, and they were confimed to be integrated in the genome of potato, Stress-related transciption factors BADH from orychophragmus, put BADH into the potato. A regeneration system available for potato transformation was established considering by several factors, such as the kind and concentration of different phytohormones and bactericides, the concentration of sucrose and agar, the time of co-culture, the effect of selection.Test to transgenic BADH of salt-resistant in potato, four transgenic plant lines compared with the control, leaf relative water content, MDA Content and Superocide Dismutase Activity, the different was significant. The main results were showed as following:1. The oplimal regeneration medium for tuber disc was MS+1mg/L IAA+2mg/L ZT.2. The optimal concentration of Agrobacterium was OD600=0.7, the optimal time of infection was 5 minutes, the effect of transformation was the best. In this condition, the rate of tuber with favourite was 90.00%, the rate of pollution was 12.50%.3. Co-culture time had great influence on callus inducation, the optimal co-culture time of favourite was 2 day.4. In the comparative experiment of two bactericides, the best one was carbenicillin made in USA, and its effective concentration was 200 mg/L, which did not have negative effect to the growth of explants.5. The manner of selection was confimed one week later, PPT was used as the selective agent. The result showed that: the concentration was 5mg/L for favourite tuber discs at the stage of shoot, 4mg/L for the root .6. Expeiment obtained 30 resistant plant , in which 10 and 4 transformants were confirmed by the test of PCR and Southern blotting to be transgenic plants. This result showed BADH gene was integrated into potato genomes.In salt resistant test, there are 4 transformants plants. The plants were used for Nacl concertration during 0.3%, 0.5%, 0.7% and 0.9%. In the 0.7%, the plants were growth normal.The results of physical detection showed: leaf relative water content 10.43% higher than that of the control , SOD activity 1.32 times higher than the contrast, controlled plant MDA content is higher than genetically modified strain 8.87%. Comprehensive above reslts, explain in 4transgenic BADH genes are already expressing, improve segregation of transgenic potato segregation of salt resistance.