Wild relatives of safflower (Carthamus tinctorius L.) may contribute genes to improve cultivated species under water stress conditions. This study was conducted to increase genetic diversity of safflower for agromorphological traits and fatty acid profiles using interspecific hybridization. Three species—C. tinctorius, C. palaestinus Eig., and C. oxyacanthus M. Bieb.—were used to develop three segregating populations of C. tinctorius × C. palaestinus (TP), C. oxyacanthus × C. palaestinus (OP), and C. oxyacanthus × C. palaestinus (TO). Seventy‐three lines, along with the three parental species, were evaluated for agromorphological traits under water stress and nonstress conditions in F3 and F4 generations during 2 yr. Fatty acid analysis of interspecific progenies showed occurrence of transgressive segregation. The results showed that mean values of seed yield for TP and TO progenies, where C. tinctorius was one of their parents, were higher than mean value of seed yield for OP that derived from the two wild species. High genetic variation was observed for seed yield, capitulum diameter, and phenological traits in interspecific populations under both moisture conditions. Indirect selection for flowering and seed yield components under water deficit conditions was most efficient to improve seed yield. Results indicated that gene introgression from wild relatives of safflower into the cultivated gene pool increased genetic variation for the measured traits and allowed identifying superior genotypes for future breeding programs.