一种根癌农杆菌介导转化平菇菌丝体的新方法

设计了一种简单高效的根癌农杆菌介导转化平菇菌丝体的方法。利用嵌套平板的方法，在内层平板培养平菇菌丝，待菌丝生长至平板边缘，将经诱导的根癌农杆菌与共培养培养基（CCM）混合，倒入内层平板与外层平板的间隙中，与正在生长中的菌丝共培养，实现转化，利用潮霉素抗性筛选获得转化子，平均转化效率33%左右。此方法操作简单，周期短，极易扩大实验规模，为平菇的分子育种以及其他丝状真菌的遗传转化提供参考。
In this paper，we developed an easy and efficient method for Agrobacterium-mediated transformation of the mycelia of Pleurotus ostreatus. Using nested plates，the mycelia of Pleurotus ostreatus were grown on the inner plate. When the mycelia grew to the edge of the plate，the induced Agrobacterium tumeifaciens were mixed with co-cultivation medium（CCM） and then were added to the space between the inner and outer plate，the above mixtures further were co-cultivated with the growing mycelia. The average conversion rate of the transformations obtained by the usage of resistance screening to the hygromycin B is about 33%. The advantages of this method are simple operation，short term，and easy to expand the scale of experiments. The results may provide reference for the molecular breeding of Pleurotus ostreatus and the genetic transformation of the other filamentous fungi