Multiplex Microsatellite Analysis Technology for Genetic Identification of Sugar Beet Lines and Hybrids

A technology for sugar beet genotyping has been developed (Beta vulgaris) at ten microsatellite loci: FDSB1001, FDSB1033, SB04, SB09, SB15, Unigene16898, Unigene17623B, Unigene17923, Unigene26753, and Unigene27833. The technology can be used for efficient, accurate, and rapid identification of sugar beet lines and hybrids. The proposed approach includes multiplex PCR for all loci in one test tube followed by electrophoretic analysis of the obtained DNA fragments in one capillary of a genetic analyzer. One of the key elements of the technology, determining the accuracy and reproducibility of the analysis, is the use at the electrophoresis stage of an additional length standard, an allelic ladder consisting of DNA fragments of the analyzed microsatellite loci. The advantages of the proposed technology include the possibility of standardization and automation of the method in a 96-well plate format, which allows for the establishment of a mass analysis process. As a result of this study, the DNA profiles of six modern sugar beet hybrids (Azimut, Visit, Korvet, Rubin, Fregat, and Uspekh) and their component lines were obtained for the first time. The developed technology can be used effectively for genetic identification of sugar beet lines, assessment of the homogeneity of plant material, and quality control of hybridization at all stages of the breeding process and can become a reliable laboratory tool for supporting industrial seed production of this crop.