马铃薯'华颂66号'遗传转化体系的建立OACSTPCD

In order to explore the optimal genetic transformation system for potatoes,the genetic transformation system of purple potato'Huasong 66',using stem segments of plantlets in vitro as explants.Plant silencing vector plasmid pBWA(V)KS-miR8019 was used to investigate the effects of different pre-culture times,plant hormones and their ratios,selection concentrations,and antibiotics using Agrobacterium tumefaciens transformation method.The results indicate that:1)The pre-culture time of stem segments was 2 d,and the callus induction ratewas the highest.2)The optimum callus induction medium was MS+2.0 mg/L 6-BA+1.0 mg/L 2,4-D and the ratio of callus induction reached 96.7％.The optimum callus differentiation medium was MS+4.0 mg/L 6-BA+O.1 mg/L NAA+1.0 mg/L GA3.3)The optimum screening pressure for plants regeneration and rooting was 75 mg/L Kan.4)The optimum concentration of agent for Agrobacterium bacteriostatic inhibition was 300 mg/L timentin,and the timentin was needed in callus induction,differentiation and rooting.In conclusion,the genetic transformation system of'Huasong 66'was preliminarily established through studying the factors at each stage of Agrobacterium-mediated genetic transformation,which would provide the certain basis for the development of potato molecular breeding.