The Genetic Transformation Of ODREB2B Gene In Potato And Identification Of Transgenic BcBCP1 Gene Of The Progenies In Potato

In this study,ODREB2B gene was transferred into Huangmazi and zhongshu3,two 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.Stress-related transcription factors ODREB2B from Orychophragmus,put ODREB2B into the 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 concentration of sucrose and agar,the time of co-culture,the effect of selection.Test to transgenic BcBCP1 of drought-resistant in potato,six transgenic plant lines compared with the control,Leaf relative water content,MDA Content and Superoxide Dismutase Activity,the difference was significant.The main results were showed as following:1.The genetic transformation of ODREB2B gene in potato1.1 The oplimal regeneration medium for tuber disc was MS+1mg/L IAA+4mg/L ZT.1.2 The optimal concentration of Agrobacterium was OD₆₀₀ 0.5,the optimal time of infection was 5 minutes,the effect of transformation was the best.In this condition,the rate of tuber disc with Huangmazi was 122.86%,the rate of tuber disc with Zhongshu3 was 106.06%.1.3 Co-culture time had great influence on callus induction rate,the optimal co-culture time of Huangmazi was 2 days,and the optimal co-culture time of Zhongshu3 was 3 days.1.4 Different types and concentrations of bactericides Comparative tests,the best one was imported Cefotaxime sodium Invitrogen,its effective concentration was 150mg/L,and it was inhibited agrobacterium strain LBA4404,which didn't have negative effect to the growth of explants.1.5 The manner of selection was confirmed one week later.PPT was used as the selective agent. The results showed that:the concentration was 5mg/L for Huangmazi and Zhongshu3 tuber discs at the stage of shoot,4mg/L for the root formation in Huangmazi and 3mg/L for the root formation in Zhongshu3.1.6 33 Experiment obtained 33 resistant plants of Huangmazi and 27 resistant plants of Zhongshu3,and in which 16 and 14 transformants were confirmed by the test of PCR and PCR-Southern blotting to be transgenic plants.This result showed BcBCP1 gene was integrated into potato genomes.2.Biological identification of transgenic BcBCP1 gene of the progenies in HuangmaziIn drought resistant test,harvest for transgenic BcBCP1 of Huangmazi in past year,6 transgenic plants and control plants of morphological indexes and physiological and biochemical indexes were stressed by drought were analyzed.After 15 days' drought stress,stems and leaves were green,leaves were out of curl of transgenic plants;Stems and leaves were yellow-green, leaves were curl of non-transgenic plants;The results of physical detection showed:leaf relative water content 14.98%higher than that of the control,SOD 27.48%higher than that of the control, MDA Content 43.83%higher than that of the control.That means BcBCP1 gene had expressed in all 6 transgenic plants,and enhanced potato transgenic plants drought resistance.