Ascorbate (vitamin C) can reach very high concentrations in chloroplasts (20–300 mM).The pool size in leaves and chloroplasts increases during acclimation to high light intensity and the highest concentrations recorded are in high alpine plants. Multiple functions for ascorbate in photosynthesis have been proposed, including scavenging of active oxygen species generated by oxygen photoreduction and photorespiration, regeneration of α–tocopherol from α–tocopheryl radicals, cofactor for violaxanthin de–epoxidase and donation of electrons to photosystem II. Hydrogen peroxide scavenging is catalysed by ascorbate peroxidase (Mehler peroxidase reaction) and the subsequent regeneration of ascorbate by reductant derived from photosystem I allows electron flow in addition to that used for CO2 assimilation. Ascorbate is synthesized from guanosine diphosphate–mannose via L–galactose and L–galactono–1,4–lactone. The last step, catalysed by L–galactono–1,4–lactone dehydrogenase, is located on the inner mitochondrial membrane and uses cytochrome c as electron acceptor. L–galactono–1,4–lactone oxidation to ascorbate by intact leaves is faster in high–light acclimated leaves and is also enhanced by high light, suggesting that this step contributes to the control of pool size by light. Ascorbate–deficient Arabidopsis thaliana vtc mutants are hypersensitive to a number of oxidative stresses including ozone and ultraviolet B radiation. Further investigation of these mutants shows that they have reduced zeaxanthin–dependent non–photochemical quenching, confirming that ascorbate is the cofactor for violaxanthin de–epoxidase and that availability of thylakoid lumen ascorbate could limit this reaction. The vtc mutants are also more sensitive to photooxidation imposed by combined high light and salt treatments.