以G418抗性为选择标记的金龟子绿僵菌原生质体转化的研究

Protoplast Transformation of Metarhizium anisopliae with G418 Resistance as Selective Marker

[目的]提高绿僵菌的生防效果。[方法]利用PEG方法,将含有G418抗性标记的质粒pSM334转化绿僵菌原生质体,并在高于绿僵菌敏感的G418浓度下筛选转化子。[结果]转化率达15~23个/μg质粒。将转化子在非选择压力下连续培养5代,其抗性稳定不变。稳定性试验表明,所获得的转化子可通过有丝分裂稳定传代。[结论]该研究为提高绿僵菌的生防效果提供了依据。

[Objective] The study aimed to enhance bio-control effect of M.anisopliae.[Method] The plasmid pSM334 containing G418 resistance marker was transformed into M.anisopliae protoplast by PEG method and the transformants were screened under the G418 concentration over the sensitive concentration of M.anisopliae.[Result] The transformation rate was 15～23个/μg plasmid.The transformants were cultured for 5 generations continuously under nonselective pressure and their resistance was stable and unchanged.The stability test indicated that the obtained transformants could passage stably through mitosis.[Conclusion] The study provided basis for increasing the bio-control effect of M.anisopliae.