The aim of this research was to gauge the alternations in the lipid peroxidation and antioxidative enzyme activity in two cultivars (cv. RGS003 and cv. Sarigol) of canola under drought stress and drought tolerance amelioration by penconazole (PEN) and calcium (Ca). Plants were treated with different polyethylene glycol (PEG) concentrations (0, 5, 10 and 15%) without or with PEN (15 mg L-1) and Ca (15 mM). The Ca treatment prevented the negative effects of drought on fresh weight (FW) in RGS003 and Sarigol at 5 and 15% PEG respectively. Ca and PEN/Ca treatments caused significant induction in the proline content in Sarigol at 15% PEG; the latter treatment was accompanied by higher glycine betaine (GB), lower malondialdehyde (MDA) and growth recovery. Hydrogen peroxide (HO2) content in Sarigol was proportional to the severity of drought stress and all PEN, Ca and PEN/Ca treatments significantly reduced the H2O2 content. PEN and PEN/Ca caused alleviation of the drought-induced oxidative stress in RGS003. RGS003 cultivar exhibited significantly higher antioxidative enzymes activity at most levels of drought, which could lead to its drought tolerance and lower MDA content. In contrast to that of Sarigol, the activity of catalase and superoxide dismutase (SOD) increased with Ca and PEN/Ca treatments in RGS003 under low stress. The application of PEN and Ca induced significantly P5CS and SOD expression in RGS003 under drought stress after 24 h. Overall, these data demonstrated that PEN and Ca have the ability to enhance the tolerance against the drought stress in canola plants.