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Citrullus lanatus (Thunb.) Matsumura and Nakai (Cucurbitaceae) is an important cucurbit crop worldwide. Global production of watermelon is about 90 million metric tonnes per annum, making it among the top five most consumed fresh fruits. The objective of this study was to evaluate seed variability in different segregating populations, and determine heritability of traits of watermelon. Interspecific crosses were made between two cultivars of C. lanatus (Bebu and Wlêwlê Small Seeds (WSS) were performed at Research Station of Nangui Abrogoua University in Abidjan, Côte d’Ivoire. There was wide variability between parental, F1, BC1 (first generation of back-crossing) and F2 seeds. Seeds of all hybrid populations were intermediate versus those of the parents. Also, crossing did not affect F1 and F2 seed characters, but affected those of BC1 because of maternal effects. Thus, back-crossing on Bebu cultivar produced seeds which looked like those of Bebu; while back-crossing on WSS cultivar produced seeds similar to those of WSS. Principal Component Analysis (PCA) and individuals repartitioning revealed that Bebu and WSS cultivars were genetically distinct and showed three main groups: two groups from each parental line and one from a recombinant line (hybrids). F2 population had a wide individual’s dispersion, and contained seeds of all other populations. High heritability was observed for all evaluated characters.