Thirty-eight melon accessions, which are of primary breeding importance in the Ukraine, were analysed for diversity. These collections represent a major non-US and non-western Europe source of melon germplasm that have not yet been subjected to molecular characterization. Molecular diversity was estimated based on a robust set of 465 polymorphisms gathered by amplified fragment length polymorphisms and simple sequence repeats (SSR). In this paper, we report 12 newly developed polymorphic SSR primer pairs, and their use for molecular characterization in the Ukrainian melon collections. Based on these polymorphisms, we estimated similarity indices that ranged from 0.70 to 1.00 among various accessions. The phylogenetic tree based on the similarity indices and a three-dimensional plot of the first three vectors of the principal component analysis corresponded fairly well with the existing three classical morphotypes namely aestivalis, europeus and hiemalis, under the convar Europeus, which is also known as adana. The polymorphisms generated in the current study, which are specific to the grouping of fruit types and days to maturity will be very useful for further genetic studies and marker-assisted selections.