Agrobacterium-mediated Transformation Of Potato With Drought Resistance Gene BcBCP1

In this study, BcBCP1 gene was transferred into Huangmazi, a 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 confirmed to be integrated in the genome of potato. A regeneration system available for potato transformation was established considering by severial factors, such as the kind and concentration of different phytohormones and bactericides, the time of co-culture. The physiological Index showed transgenic potato plants are with strong tolerance to drought stress.The main results were showed as following:1 The oplimal regeneration medium for tuber disc was MS+4mg/L ZT + 1mg/L IAA. The highest rate of inducing shoot is 86%.2 For BcBCP1 gene, the optimal concentration of Agrobacterium was OD600 0.5, the optimal time of infection was 5 minutes. In this condition, the rate of tuber disc with Huangmazi was 87.19%, the rate of pollution was 1.2%.3 Co-culture time had great influence on callus induction rate, The optimal co-culture time of Huangmazi with BcBCP1 gene was 2 days, In this condition, the rate of tuber disc with was 83.97%, the rate of pollution was 6.77%.4 In the comparative experiment of two bactericides, the effective concentration with Cefotaxime sodium made in China was 400mg/L. The best one was Cefotaxime sodium made in USA, and its effective concentration was 150mg/L , which didn't have negative effect to the growth of explants.5 In this study, PPT was used as the selective agent. The manner of selection was confirmed 7 days later.The concentration was 4mg/L for the tuber discs at the stage of shoot ,and 5mg/L for the root formation.6 72 transformants were attained for BcBCP1 gene in this experiment, in which 35 plants were passed rootage (PPT 5mg/L), and in which14 transformants were confirmed by the test of PCR testing. Southern blot showed that 6 transformants had hybridization signals. This results showed BcBCP1 gene were integrated into potato genomes.7 In drought resistant test, 6 transgenic plants and control plants were stressed by drought and the physiological index were analyzed. The results showed: after 15 days'drought stress, the relative water content of 4 transgenic plants and the control plants had achieved notable difference level. The increasing content of proine in control plants was 80.20%, and in 6 transgenic plants was 181.63%-396.28%; At the same time, the chlorophyll content of 6 transgenic plants and control plants were measured ,and result showed the decreasing rate of chlorophyll content was 43.73% in control plant, while the number is 21.98%-32.62% in transgenic plants. That means BcBCP1 gene had expressed in all 6 transgenic plants, and enhanced'Huangmazi'transgenic plants drought resistance.