Magnesium aminoclay-mediated genetic transformation of Trichoderma atroviride HB20111

Abstract:

[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 10⁶ 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/cm²,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.

Key words:magnesium aminoclay;Trichoderma atroviride HB20111;ultrasound;green fluorescent protein