In Ukraine, the area under garlic is increasing and as of 2020 amounted to 1,100 hectares, due to the high profitability of production and constant demand in both domestic and foreign markets. At the same time, the evaluation of existing and new varieties of winter garlic is important for further growth in yields of this important crop. The aim of the research is to assess the genetic diversity of the studied varieties of winter garlic by ISSR loci and to determine the most productive in the conditions of the Left Bank Forest-Steppe of Ukraine. Winter garlic varieties Duchess, Lyubasha and Hungarian (line 20-16) were used for the research. The highest level of marketable yield (14.1 t / ha) was obtained when growing winter garlic of Lyubasha. The Hungarian variety (Line 20-16) provided a yield of 12.9 t / ha, the Duchess variety 9.4 t / ha. The formation of winter garlic yield by 12.3 % depended on the characteristics of the variety, the influence of the growing season was 87.3 %. Agronomically stable (As ˃ 70 %) was the winter garlic variety Hungarian (Line 20-16) with As = 73.8 %. Based on the above studies, allelic formulas (molecular genetic passports) of winter garlic varieties Hungarian (Line 20-16), Lyubasha and Duchess were formed. In the studied varieties of winter garlic, several amplicons were found, unique within the study group. The UCC812725 and UBC 812902 loci were unique in the Duchess variety, and the UBC 812791 and UBC 842702 loci were unique in the Lyubasha variety. The calculated Nei–Li similarity coefficients indicate a significant genetic similarity of the studied varieties of winter garlic. The Lyubasha and Duchess varieties are genetically closer to each other than the Hungarian varieties (Line 20-16).