SET OF OLIGONUCLEOTIDE PRIMERS FOR GENOTYPING OF FUNGI OF COLLETOTRICHUM ACUTATUM SPECIES COMPLEX Russian patent published in 2023 - IPC C12Q1/68 

FIELD: biochemistry; biotechnology; molecular biology.

SUBSTANCE: following is proposed: a method of determining the species identity of isolates and/or strains of fungi of the species complex Colletotrichum acutatum in a sample, and the said method includes the following steps: extracting total DNA from a sample presumably containing fungal mycelium; carrying out PCR with primers and the nucleotide sequences of the amplicons of ITS, GAPDH, TUB2, APN/MAT1 loci are determined by sequencing; using the method of sequence alignment and phylogenetic comparison with genetic databases containing sequences for the ITS, GAPDH, TUB2, APN/MAT1 loci for fungi of the species complex Colletotrichum acutatum, the qualitative species and intraspecific composition of fungi of the genus Colletotrichum in the analyzed sample are determined.

EFFECT: invention makes it possible to expand the set of tools for amplification and targeted sequencing of a new generation of fragments of loci (ITS, GAPDH, TUB2, APN/MAT1) of various isolates of the Colletotrichum lupini, the causative agent of anthracnose white lupine and yellow lupine, as well as related species of the Colletotrichum acutatum complex (C. tamarilloi, C.costaricensis, C.melonis, C.cuscutae, C.paranaense, C.nymphaeae, C.simmondsii, C.acutatum); the invention also makes it possible to increase the accuracy and speed of establishing the species identity of monosporous and multisporous isolates of species of the Colletotrichum acutatum complex, allows to study their phylogenetic relationship at the species level and identify agriculturally significant anthracnose isolates, phytosanitary control of the spread of pathogenic strains, control the infectious background in breeding and seed-growing areas of white and white lupine yellow lupine.

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