METHOD FOR IDENTIFYING IMPORTANT FOR AGRICULTURE FUNGI OF THE GENUS TRICHODERMA-TRICHODERMA HARZIANUM, TRICHODERMA VIRIDE AND TRICHODERMA ATROVIRIDE BASED ON PCR-RFLP Russian patent published in 2024 - IPC C12Q1/68 

FIELD: microbiology.

SUBSTANCE: described is a method for molecular genetic identification of members of the genus Trichoderma, such as T. harzianum, T. viride and T. atroviride. DNA is recovered from the pre-collected biological material. PCR, restriction of PCR products, analysis of polymorphism of length of restriction fragments (RFLP) of amplicon, electrophoresis in agarose gel are carried out. PCR is carried out using primers: direct ITS1 SEQ ID NO: 1 and reverse ITS4 SEQ ID NO: 2. Restriction enzyme Bam HI is used for identification during restriction analysis, which forms unique restriction fragments for three fungi of the genus Trichoderma—T. harzianum, T. viride and T. atroviride, where the presence of DNA fragments with length of 430 bps and 178 bps indicates belonging to a type of fungi T. atroviride, with length of 520 bps and 119 bps—for belonging to a type of fungi T. viride, length of 620 bps—for belonging to the type of fungi—T. harzianum.

EFFECT: identification of fungi of Trichoderma harzianum, Trichoderma viride and Trichoderma atroviride species without involvement of specialists in microbiology by standardized methods of molecular genetics, with possibility of identification procedures for 4,5–6 hours.

1 cl, 1 dwg, 1 ex