Технология генетической идентификации сортов и диких форм винограда на основе мультилокусного микросателлитного анализа

Abstract

The genotyping technology has been developed based on 9 microsatellite loci (VVS2, VVMD5, VVMD7, VVMD27, VVMD28, VVMD25, VVMD32, VrZAG62, VrZAG79). It can be used for efficient, accurate and fast identification of grape varieties and forms (genus Vitis). The proposed approach includes a multiplex PCR of all loci followed by electrophoretic analysis of DNA fragments in one capillary of a genetic analyzer. The application of an additional length standard, an allelic ladder, consisting of all possible DNA fragments of the analyzed microsatellite loci, is one of the key features of the technology, which ensures the accuracy and reproducibility of the results. The advantage of the proposed technology is the ability to standardize and automate the procedure using 96-well plates, which opens up the possibility of conducting mass analyses. As a result of the study of varieties and forms of Vitis genus species, genetic passports were obtained, according to which a dendrogram was constructed, reflecting the genetic relationship of the studied samples. The developed technology makes it possible to distinguish varieties and wild forms of grapes; it can be used for their identification and determination of genetic distance between them, as well as for assessment of planting material and protection of breeders' rights. Vitis, grapes, microsatellites, SSR, genotyping, multiplex PCR, DNA fragment analysis The work was carried out within the framework of the state assignment on the topic "Development of New Technologies for Genetic Analysis of Forms of Agricultural Plants to Accelerate and Control the Selection Process" (project no. 0574-2019-0003).