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The mechanisms controlling the photosynthetic performance of C4 plants at low temperature were investigated using ecotypes of Bouteloua gracilis Lag. from high (3000 m) and low (1500 m) elevation sites in the Rocky Mountains of Colorado. Plants were grown in controlled-environment cabinets at a photon flux density of 700 μmol m−2 s−1 and day/night temperatures of 26/16 °C or 14/7 °C. The thermal response of the net CO2 assimilation rate (A) was evaluated using leaf gas-exchange analysis and activity assays of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), phosphoenolpyruvate carboxylase (PEPCase) and pyruvate,orthophosphate dikinase (PPDK). In both ecotypes, a reduction in measurement temperature...