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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: To realize the genetic transformation of Trichoderma sporeatum T4 and optimize its transformation system. Methods: The hygromycin resistant marker was selected and transformed into Trichoderma sp. By Agrobacterium tumefaciens transformation. Results: The hygromycin gene was successfully integrated into the recipient bacterial genome, and the transformant resistance gene could be stably inherited. Conclusion: The optimal transformation system and conditions were AS concentration of 200μg / mL in IM and CM medium, 106 / mL of spore of Trichoderma sp., 200μL of Agrobacterium tumefaciens (OD600 about 0.8), co-culture time For 48 h, the transformation efficiency was about 50 transformants / 106 spores.