In a previous study, we found that enhanced CO2 subjected to nodulated alfalfa plants grown at different temperatures (ambient and ambient + 4 degrees C) and water availability regimes could protect PSII from photodamage. The main objective of this study was to determine the mechanism(s) involved in the photoprotection of PSII at elevated CO2 levels in this plant. Elevated CO, reduced carboxylation capacity-induced photosynthetic acclimation and reduced enzymatic and/or nonenzymatic antioxidant activities, suggesting that changes in electron flow did not cause any photooxidative damage (which was also confirmed by H2O2 and lipid peroxidation analyses). Enhanced nonphotochemical quenching and xanthophyll cycle pigments revealed that plants grown at 700 mu mol mol(-1) CO2 compensated for the reduction in energy sink with a larger capacity for nonphotochemical dissipation of excitation energy as heat, i.e., modulating the status of the VAZ components. Elevated CO2 induced the de-epoxidation of violaxanthin to zeaxanthin. facilitating thermal dissipation and protecting the photosynthetic apparatus against the deleterious effect of excess excitation energy. (C) 2008 Elsevier B.V. All rights reserved.