Wheat (Triticum aestivum L. cv. HD 2285) was grown in control (C) and heated (H) open top chambers (OTCs) for entire period of growth and development till maturity. The mean maximum temperature of the entire period was 3 °C higher in H-compared to C-OTCs. Net photosynthetic rate (PN) measured at different temperature (20–40 °C) of C-and H-grown plants showed greater sensitivity to high temperature in H-plants. PN measured at respective growth temperature was lower in H-compared to C-plants. The CO2 and irradiance response curves of photosynthesis also showed lesser response in H-compared to C-plants. The initial slope of PNversus internal CO2 concentration (PN/Ci) curve was lower in H-than C-plants indicating ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) limitation. In irradiance response curve, the plateau was lower in H-compared to C-plants which is interpreted as RuBPCO limitation. RuBPCO content in the leaves of C-and H-plants, however, was not significantly different. Ribulose-1,5-bisphosphate carboxylase (RuBPC) initial activity was lower in H-plants, whereas activity of fully activated enzyme was not affected, indicating a decrease in activation state of the enzyme. This was further substantiated by the observed decrease in RuBPCO activase activity in H-compared to C-plants. RuBPCO activase was thus sensitive even to moderate heat stress. The decrease in PN under moderate heat stress was mainly due to a decrease in activation state of RuBPCO catalysed by RuBPCO activase.