In the present study, low temperature-dependent physiological changes were investigated through photosynthetic activity and some endogenous mechanisms in two winter oilseed rape cultivars (Brassica napus L. ssp. oleifera cvv. Eurol and Hansen) on the basis of leaf age. Chlorophyll fluorescence measurements demonstrated that low temperature caused decreased photosynthetic activity in both cultivars. However, photosynthetic apparatus in the young leaves of Hansen is more tolerant to low temperature as demonstrated by lower F0 (minimum fluorescence yield) and 1-qp (excitation pressure of photosystem II), higher Fm (maximum fluorescence yield), Fv/Fm and non-photochemical quenching (NPQ) compared with Eurol. In addition, young leaves of Hansen represented marked increase in some antioxidant enzyme activities (superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR)) during cold exposure. In the young leaves of Eurol, however, APX and GR activity was decreased by low temperature, indicating lower efficiency of ascorbate-glutathione cycle. Lower antioxidant activity in the young leaves of Eurol may be responsible for increased malondialdehyde (MDA), H2O2 and membrane damage and decreased chlorophyll content as a result of oxidative damage during cold exposure. In the cold-stressed mature leaves, both cultivars represented similar antioxidant capacities and photosynthetic efficiency. As a consequence, coordinated increase in SOD, APX and GR activities, increased capacity to keep quinone A (QA) in an oxidised state (as indicated by lower 1-qp) and accumulation of soluble sugar and proline could be mainly attributed to higher level of tolerance of the young leaves of Hansen to low temperature when compared with Eurol.