Drought stress causes various physiological and biochemical effects in plants. Chicory (Cichorium intybus L.), of the Asteraceae, is a medicinal herbal that is found in most parts of Iran. This study was taken as a factorial experiment based on a completely randomized design with four replications, to evaluate the effect of water deficit on changes in antioxidant enzyme activity, proline content, and ABA accumulation of four ecotypes of chicory (Sefid Shiraz, Siyah Shiraz, Sefid Isfahan, Siyah Isfahan). To conduct this research, greenhouse cultivation at four different irrigation levels, 100% (as a control), 80, 60 and 40% of field capacity (FC), was conducted and after 60days of tension under specific treatment, the plants were harvested and the indicators were investigated. The results of the present study indicate that Siyah Shiraz performs better than the others under water deficit conditions, as indicated by higher antioxidant enzyme (superoxide dismutase, catalase, and peroxidase) activity, proline content (as non-enzymatic anti oxidant) and abscisic acid accumulation in leaves. Since activation of the antioxidant system helps the plants with stress-induced damages, our results indicate that Siyah Shiraz has a better defense system against oxidative damage. These data are useful for better understanding of the physiological basis of changes in drought resistance as well as crop breeding projects.