Fagopyrum represents an economically and nutritionally important crop which belongs to the group of pseudocereals. Buckwheat is referred to be a functional crop with a wide range of use around the world. Grains of buckwheat are nutritionally significant. They are characterised by a high content of starch, proteins, flavonoids and fibre. SCoT technique is used for the detection of plant genes polymorphism, for the evaluation of genetic diversity and for mapping the functional regions of the genome. The aim of the work was to perform molecular analyses of 21 genotypes of Fagopyrum esculentum Moench and 14 genotypes of Fagopyrum tataricum Gaertn. using ten gene-targeted SCoT markers. The seedlings of buckwheat were used for DNA isolation and subsequently for the amplification of the DNA using a polymerase chain reaction. The number of amplified fragments ranged from 12 (SCoT 60, SCoT 13) to 27 (SCoT 12). The total number of fragments was 176 of which 162 were polymorphic with an average of polymorphic fragments 16.2. The percentage of polymorphism ranged from 58.33 % (SCoT 60) to 100 % (SCoT 12, SCoT 13, SCoT 29, SCoT 30) with an average of 90.29 %. Polymorphic information content (PIC) values characterize the polymorphism of used SCoT markers. The PIC values ranged from 0.578 (SCoT 60) to 0.932 (SCoT 36) with an average of 0.859. The genetic diversity of buckwheat was determined by hierarchical cluster analysis using the UPGMA algorithm in the created dendrogram. Genotypes of buckwheat were divided into two main clusters. Two genotypes of common buckwheat (Siva, Špačinska I) from Slovenia and the Slovak Republic, respectively, as well as another two tartary buckwheat genotypes (Tohno Zairai, Winsor Royal) from unknown regions, respectively, were genetically the closest. The PCoA plot confirmed the separation of tartary buckwheat from common buckwheat resulting in the constructed dendrogram According to our results, we can consider the SCoT technique appropriate for differentiation of Fagopyrum esculentum Moench and Fagopyrum tataricum Gaertn. genotypes leading to genotype identification and utilization in the breeding process to improve the buckwheat genetic material.