镁硅酸盐纳米粘土介导的深绿木霉HB20111的遗传转化  

Magnesium aminoclay-mediated genetic transformation of Trichoderma atroviride HB20111

作　　者：隋丽娜 扈进冬 吴远征 魏艳丽 李纪顺 王燕[1] SUI Lina;HU Jindong;WU Yuanzheng;WEI Yanli;LI Jishun;WANG Yan(School of Bioengineering,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250353,Shandong,China;Shandong Provincial Key Laboratory of Applied Microbiology,Ecology Institute,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250103,Shandong,China)

机构地区：[1]齐鲁工业大学(山东省科学院)生物工程学院,山东济南250353 [2]齐鲁工业大学(山东省科学院)生态研究所山东省应用微生物重点实验室,山东济南250103

出　　处：《微生物学通报》2022年第3期956-963,共8页Microbiology China

基　　金：山东省重大科技创新工程项目(2020CXGC010803,2019JZZY010718);齐鲁工业大学(山东省科学院)校(院)地产学研协同创新基金项目(2020-CXY42);齐鲁工业大学(山东省科学院)科教产融合创新试点工程项目(2020KJC-GH07);山东省科学院国际科技合作项目(2019GHPY05)。

摘　　要：【背景】木霉菌遗传转化方法的烦琐复杂限制了对其基因克隆、基因功能的研究。【目的】建立深绿木霉HB20111便捷、高效的遗传转化体系。【方法】利用镁硅酸盐纳米粘土作为载体吸附含有荧光蛋白基因gfp的丝状真菌表达载体pCAMBIA1303-gpdA-GFP-TrpC-Hygro,形成质粒-镁硅酸盐纳米粘土复合物,在超声条件下对深绿木霉HB20111分生孢子进行遗传转化。【结果】当分生孢子浓度为10^(6)CFU/mL、分生孢子悬液培养12h、镁硅酸盐纳米粘土浓度100mg/L和超声处理30 s(输出功率为100 W/cm^(2),发射频率为50 kHz)条件下对深绿木霉HB20111的转化效率最高,可以达到124个转化子/μg-DNA。【结论】使用镁硅酸盐纳米粘土作为载体可以实现对深绿木霉HB20111便捷、高效的遗传转化,转化子抗性基因和报告基因可以稳定遗传。[Background]The complicated genetic transformation methods hinder the gene cloning and gene function characterization of Trichoderma.[Objective]To establish a convenient and efficient genetic transformation system for Trichoderma atroviride HB20111.[Methods]Magnesium aminoclay was used as a carrier to adsorb the filamentous fungal expression vector pCAMBIA1303-gpdA-GFP-TrpC-Hygro containing the fluorescent protein gene gfp to form a plasmid-magnesium aminoclay complex.The conidia of T.atroviride HB20111 were then genetically transformed under ultrasonic conditions.[Results]Under the conditions of the conidial suspension at a concentration of 10^(6) CFU/mL,culture time of 12 h,magnesium aminoclay concentration of 100 mg/L,and ultrasonic treatment for 30 s(the ultrasonic output power was 100 W/cm^(2),and the emission frequency was 50 kHz),the transformation efficiency of HB20111 was the highest,which reached 124 CFU/μg-DNA.[Conclusion]Using magnesium aminoclay as a carrier can realize convenient and efficient genetic transformation of T.atroviride HB20111,and the resistance gene and reporter gene can be inherited stably in the transformant.

关 键 词：镁硅酸盐纳米粘土 深绿木霉HB20111 超声 绿色荧光蛋白 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]