Drought is a severe environmental constraint to growth and production of crop plants world wide. Wild species of cultivated plants carry useful genes which can be used for breeding programs. To evaluate drought tolerance and especially traits related to root in cultivated and wild safflower genotypes, this experiment was conducted in two years and the experimental design was split plot arrangement in RCBD with 3 replications. The levels of irrigation that were considered as the main factor consisted of normal condition that was based on the depletion of 60% of soil moisture content and moderat stress with 75% and severe stress with 50% of water requierments for normal condition. In the first year, genotypes as a subsidiary factor were consisted of Kashan, C4110, Azari, Kermanshah and the genotype derived from inter specific cross and in second year the p6 genotype from palaestinus spicies was added. Phenologic and agronomic traits, charestricteristics of aerial plant parts and root, seed yield and yield compenent and proline content in two years, membrane stability index (MSI) and phenol content in the first year and relative water content, activity of antioxidant enzymes in second year were measured. There was a considerable variation among wild and cultivated genotypes and the genotype getting of cross in normal and stress conditions for all studied traits. Cultivated genotypes passed phonologic stages earlier than wild genotypes.In the first and second year, moisture stress had a significant effect on seed yield. Kashan and Kermanshah genotypes had minimum percentage of reduction of seed yield in stress condition in the first and second year respectively. Drought stress had not significant effect on root length, however, increasing water stress level caused increase in the length of root that the most presentage of its increase belonged to cultivated genotypes in two years. Drought stress caused significant increase in proline content so that in the first year oxyacanthus species had more proline than tinctorius and the most percentage of increase in second year was related to p6 genotype. Activity of antioxidant enzymes was increased significantly with increasing water stress levels. In severe stress condition, the activity of peroxidase and ascorbat peroxidase enzymes were increased in oxyacanthus spcies and p6 genotype ( C. palaestinus ) more than other genotypes respectively. Genotype from cultivated species and Kermanshah genotype had the maximum increase of catalas enzyme activity. The most pecentage of phenol content was showed in kashan genotype. Relative water cotent deacreased in stress condition and the least percentage of reduction was revealed in genotype getting of specific cross. On the other hand, MSI was increased in this genotype in severe stress, This genotype hasbeen drived from a cross between wild genotype of Isfahan and genotype C4110 from cultivated species, and had the most persent of increase in proline content in severe water stress. In the first and second year, under moderate water stress, Mean productivity (MP) was superior index for identification tolerant genotypes. In severe water stress selection based on geometric mean productivity (GMP) and stress tolerance index (STI) corresponded to diserable seed yield and low sensitivity to water stress . In the first year, in mild water stress based on MP index, kashan genotype and the genotype derived from cross had the most tolerance and C4110 genotype was introduced as the most sensitive and in severe stress based on GMP and STI indices, kashan genotype was introduced as the most tolerant and similarity to mild stress, C4110 genotype was the most sensitive. In second year, based on MP index in mild stress p6 and Azri genotypes were introduced as the most tolerant and the most sensitive respectively, and based on STI and GMP indices, Kermanshah was the most tolerant and Azri had the lowest sensivity similar to mild stress. Gentype derived from cross and p6 genotype showed the most percent of increase in proline content but in severe stress, increased sensivity of this genotypes to drought stress was observed, It means that high accumulation of prolin helped these genotypes just to survive. In the first year, kashan genotype had the most percent of increase in root length under drought stress condition. Apparently for this reason, kashan genotype was the most tolelerant. Reduction of growth period in C4110 genotype caused intense decrease in seed yield, so it was introduced as the most sensitive genotype. In second year, it seems that more percent of increase in antioxidant enzyme systems and low reduction of RWC in Kermanshah genotype as compared to cultivated and p6 genotypes had key role for tolerance to drought stress. As a whole, results showed that physiological mechanisms especially antioxidant enzymes systems in wild species were more active than cultivated species.