Cotton (Gossypium hirsutum L., cv DPL 5415)) plants were grown in naturally lit environment chambers at day/night temperature regimes of 26/18 (T-26/18)), 31/23 (T-31/23)) and 36/28 °C (T-36/28)) and CO2 concentrations of 350 (C-350)), 450 (C-450)) and 700 μLL−1 (C-700)). Net photosynthesis rates, stomatal conductance, transpiration, RuBP carboxylase activity and the foliar contents of starch and sucrose were measured during different growth stages. Net CO2 assimilation rates increased with increasing CO2 and temperature regimes. The enhancement of photosynthesis was from 24 μmol CO2 m−2 s−1 (with C-350 and T-26/18)) to 41 μmol m−2 s−1 (with C-700 and T-36/28)). Stomatal conductance decreased with increasing CO2 while it increased up to T-31/23 and then declined. The interactive effects of CO2 and temperature resulted in a 30% decrease in transpiration. Although the leaves grown in elevated CO2 had high starch and sucrose concentrations, their content decreased with increasing temperature. Increasing temperature from T-26/18 to 36/28 increased RuBP carboxylase activity in the order of 121, 172 and 190 μmol mg−1 chl h−1 at C-350, C-450 and C-700 respectively. Our data suggest that leaf photosynthesis in cotton benefited more from CO2 enrichment at warm temperatures than at low growth temperature regimes.