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目的:建立籼型杂交稻亲本V20B成熟胚愈伤组织培养的高效遗传转化体系。方法:以V20B的成熟胚作为外植体,研究比较不同培养基、不同生长物质类型及浓度、不同培养条件对愈伤组织诱导、继代、分化的影响。结果:诱导培养基以N6为基本培养基,添加2.5 mg.L-12,4-D和0.2 mg.L-16-BA,光照条件下诱导,成熟胚愈伤组织诱导率达到93.44%;继代培养基以MS为基本培养基,添加2.0 mg.L-12,4-D,黑暗条件下培养;分化培养基以DL为基本培养基,激素配比为2.0 mg.L-1KT、2.0 mg.L-16-BA、0.2mg.L-1NAA、0.2 mg.L-1IAA;采用农杆菌介导法对该体系获得的愈伤组织侵染后能获得的抗性愈伤组织,经PCR检测潮霉素基因转化率为53.89%。结论:建立了适宜于籼型稻V20B的高效组织培养体系。
Objective: To establish a high efficient genetic transformation system for callus induction of mature embryos of V20B in indica hybrid rice. Methods: The mature embryos of V20B were used as explants to study the effects of different culture media, different growth media types and concentrations, and different culture conditions on the induction, subculture and differentiation of callus. Results: The induction medium was N6, 2.5mg.L-12,4-D and 0.2 mg.L-16-BA were added into the induction medium, and the induction rate of mature embryo callus reached 93.44% Substitute medium with MS as the basic medium, adding 2.0 mg.L-12,4-D, dark conditions; differentiation medium with DL as the basic medium, the hormone ratio of 2.0 mg.L-1KT, 2.0 mg L-16-BA, 0.2mg.L-1NAA and 0.2 mg.L-1IAA respectively. The resistant callus obtained by the system after Agrobacterium-mediated transformation was detected by PCR Hygromycin gene conversion was 53.89%. Conclusion: An efficient tissue culture system suitable for indica rice V20B was established.