农杆菌介导生防棘孢木霉菌转化体系的优化

Optimization of Trichoderma asperellum Transformation System with Agrobacterium tumefaciens

目的:实现棘孢木霉菌T4的遗传转化并优化其转化体系。方法:以潮霉素抗性为选择标记,利用农杆菌转化法介导转化棘孢木霉菌。结果:潮霉素基因成功整合到受体菌基因组中,转化子抗性基因可稳定遗传。结论:最优的转化体系和条件为:IM和CM培养基中AS浓度为200μg/mL,棘孢木霉T4孢子浓度为106/mL,农杆菌浓度为200μL(OD600约0.8),共培养时间为48 h,转化效率约为50个转化子/106个孢子。

Objective:The filamentous fungi Trichoderma asperellum was transformed and the conditions were optimized.Method:Agrobacterium genetic transformation was applied in Trichoderma asperellum T4 and with the selection markers for hygromycin B resistance.Result:The hygromycin B gene had been inserted into the genome of Trichoderma asperellum T4,indicating that the resistance gene is stable genetically.Conclusion:Under the optimized conditions,the concentration of AS in IM and CM medium was 200μg/mL,the spores of T.asperellum was 106/mL,and the concentration of A.tumefaciens was 200μL(OD600=0.8),48h in co-culture medium,which resulted in a reasonable ATMT rates.About 50 transformants /106.