The Technology of Genetic Identification of Varieties and Wild-Growing Forms of Grapes Based on Multilocus Microsatellite Analysis

• Published in: Applied biochemistry and microbiology Vol. 58; no. 9; pp. 1050 - 1059
• Main Authors: Sekridova, A. V., Shilov, I. A., Kislin, E. N., Malyuchenko, O. P., Kharchenko, P. N.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 01-12-2022; Springer Nature B.V
• Subjects: Biochemistry; Biomedical and Life Sciences; Control; Deoxyribonucleic acid; DNA; Fragments; Genetic distance; Genetic relationship; Genotyping; Grapes; Life Sciences; Medical Microbiology; Metrology; Microbiology; Passports; Standardization; Vitaceae; microsatellites; multiplex PCR; SSR; genotyping; grapes; DNA fragment analysis
• ISSN: 0003-6838; 1608-3024
• DOI: 10.1134/S0003683822090083

A genotyping technology has been developed based on nine microsatellite loci (VVS2, VVMD5, VVMD7, VVMD27, VVMD28, VVMD25, VVMD32, VrZAG62, and VrZAG79). It can be used for efficient, accurate, and fast identification of varieties and forms of
Vitis
grapes. The proposed approach includes 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 possibility of standardizing and automating the procedure using 96-well plates, which opens the possibility of conducting mass analyses. As a result of the study of the varieties and forms of the
Vitis
genus species, genetic passports were created, 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-growing forms of grapes; it can be used for their identification and determination of the genetic distance between them, as well as for assessment of planting material and protection of breeders’ rights.