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Not AvailableOne of the most widely applicable and used markers considered in plant research is simple sequence repeats (SSRs). The Gy14 cucumber genome provided information for the development of SSR marker in this species which might be used in other related species additionally. We, therefore, developed genome-wide SSR markers from cucumber, which were used for genetic diversity analysis in 16 different cucurbits species and comparative mapping with melon and watermelon. A total of 2171 SSR motifs were identified from seven cucumber chromosomes. Out of them, 1749 markers belong to melon while 442 markers to watermelon. We experimentally validated 70 SSR markers to check their cross-species transferability among 16 different cucurbits species belongs to six genera. Maximum% polymorphism has been recorded for Momordica charantia (82%) followed by Momordica cochinchinesis (78.18%) and Luffa harmaphrodita (73.68%) while Benincasa hispida (49.21%) exhibited minimum% polymorphism. The large number of cucumber SSR markers developed in the present study provides a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection (MAS) in cucumber. Furthermore, the cross-species transferable SSR markers could also be useful in various molecular markers related studies in other closely related species in the Cucurbitaceae family in which draft genomes are not yet available.Not Availabl