Eight sesame (Sesamum indicum L.) genotypes were evaluated using a randomized complete block design in field experiment with three replications for their diversity in seven agronomic and morphological traits at Ismailia Agricultural Research Station, ARC, Ismailia Governorate, Egypt during the two successive seasons 2016 and 2017. Results showed that the most promising genotypes for seed yield and its attributes were Zahar₁₄, Shandwell₃, H₈₈A_2, Zahar_18, B_4-2, andZahar₁₈ For all traits, estimates of phenotypic coefficient of variation (PCV) were slightly higher than those of genotypic coefficient of variation (GCV). High heritability coupled with high genetic advance (as % of the mean) was recorded for all studied traits. Five ISSR primers were used for fingerprinting of the eight sesame genotypes produced 29 bands, 23 of them were polymorphic with 79% polymorphism, while primer ISSR 1 produced two positive markers in shandweell 3 and line M_4-55. Primer ISSR 6 and ISSR 8 produced one band positive marker in the same line M_4-55.