目的:探讨根癌农杆菌介导转化平菇菌丝体的最佳转化条件。方法:以平菇ACCC52857为受体,利用潮霉素B抗性基因作为筛选标记,建立根癌农杆菌介导的平菇菌丝遗传转化体系;探讨乙酰丁香酮(AS)的浓度,根癌农杆菌的浓度,共培养的时间和共培养的温度这四个条件对转化的影响。结果:根癌农杆菌菌液浓度为OD600=0.6~0.7,诱导培养基中添加200μmol/L AS,25℃共培养3~5d,其转化效率最高。结论:对根癌农杆菌介导平菇菌丝转化体系条件的优化,为提高遗传转化效率,以及菌株选育工作奠定基础。
Objective:Optimization of the Agrobacterium- mediated transformation condition of Pleurotus ostreatus mycelia is studied. Methood:Based on earlier experiments, Pleurotus ostreatus ACCC 52857 was used as the host strain and Hygromycin B resistant gene was used as selective marker, and the transformation system of ATMT was established. This article explores the impact for the transformation from four conditions, including the concentration of acetosyringone (AS) and Agrobacterium tumefaciens, co - culture temperature and co - culture time. Result: The best transformation condition is the Agrobacterium tumefaciens liquid concentration for OD600 = 0.6 - 0.7, induction medium adding 200μmol/L AS, co - culture 3 -5 d in 25 ℃. Conclusion:The optimization of the transformation system of ATMT conditions, lay a solid foundation for improving heredity transformation efficiency and strains breeding.