Safflower ( Carthamus tinctorius L.) is one of the oldest domesticated crops, mainly grown as oilseed in the arid and semiarid regions of the world. Cross-ability has made wild Carthamus species a suitable gene source for further improvement of cultivated species. To investigate the effects of genome integration between cultivated and wild relatives of Carthamus species, three studies were attempted. In the first experiment several characteristics in several genotypes of C. tinctorius , C. palaestinus , C. oxyacanthus , C. lanatus and C. glaucus were investigated under normal and water stress condition and suitable genotypes for interspecific hybridization were selected. The second study consisted of generation mean analysis in different progeny generations derived from interspecific crosses under both water stress and non-stress conditions. In the third study Bulk Segregant Analysis was performed for head diameter in one of the interspecific crosses. The results of first study showed that speciediffered in respect to their oil content and seed yield but oil content and fatty acid profile were highly stable over different environments for all studied species. Evaluation of seed germination rate and cardinal temperatures showed a relatively similar response among the cross-able species of C . tinctorius , C. palaestinus , C. oxyacanthus under different moisture and temperature conditions. It is therefore highly probable that interspecific crosses of these species may not change oil and seed germination characteristics in the hybrids. On these bases one genotype from each of C . tinctorius , C. palaestinus and C. oxyacanthus were selected and crossed for development of F1, F2, F3, BC1:1, BC2:1 and BC1:1S1 generations. In the second study, F1 heterosis, transgressive segregation and higher drought tolerance than the parental genotypes were detected in the segregated generations. No epistatic effects were observed for most of the studied traits. Higher similarity of back cross generations to the cultivated parental genotype was evident and proved through generation mean and variance analysis. However, genetic parameters and heritability calculation will provide useful information for safflower breeders although non-normal distribution of F2 and F3 progenies, high similarity of F1 and back cross generations to cultivated parent and low environmental variance of all generations showed the complex inheritance of traits in interspecific crosses. In the third study, the three species of C. tinctorius , C. palaestinus and C. oxyacanthus were evaluated for SSR markers polymorphism. On the bases of significantly higher SSR polymorphism between C. tinctorius and C. oxyacanthus the Bulk Segregant Analysis (BSA) for head diameter was performed on the F2 generation of this cross. The results presented three SSR markers that were very likely associated with safflower head diameter. Assessment of SSR marker distribution in F2 progenies used for BSA, revealed extreme segregation distortion which helped a better interpretation of results obtained from generation mean analysis. Keywords: Safflower, Carthamus species, interspecific cross, generation mean analysis, Bulk segregant analysis.