Abstract: An investigation was conducted to study the effect of water stress on the antioxidant content, protective enzyme activities, proline content and lipid peroxidation in wheat seedlings. Drought stress increases the amount of Reactive Oxygen Species (ROS), leading to metabolic disorders. It is now known that higher levels of activity- protective mechanisms render the cells more enduring against environmental stress including drought. Two widely cultivated cultivars of wheat in Iran, Sab. and N. Sar. were grown up according to the hydroponic method. Having reached the stage of 4-5 leaves growth; the plants were kept under 4, 8 and 12 bars potential resulting from using Polyethylene Glycol 8000 (PEG 8000). Hogland solution was used as the control. Then the amount of ascorbate, glutathione, superoxide dismutase and catalase activity, proline and lipid Peroxidation was measured in cut samples of the leaves. The result indicated an increase in the amount of Ascorbate and Glutathione as the stress was intensified in the case of Sab. Moreover, the reduced form of Ascorbate (ASC) and Glutathione (GSH) were higher in Sab. at 8 and 12 bars. The amount of Proline accumulation was considerably higher in Sab. than N. Sar. SOD activity, on the other hand, diminished at 8 and 12 bar levels. CAT activity is also regarded as a limiting factor. Lipid peroxidation was also geared up as the stress was intensified. These limiting factors rendered N. Sar. cultivar more sensitive to water stress resulting from PEG8000 compared to Sab.