Colorado 80309-0334 l h e protective role of leaf antioxidant systems i n the mechanism of plant acclimation to growth irradiance was studied in Vinca major, Schefflera arboricola, and Mahonia repens, which were grown for several months at 20, 100, and 1200 pmol photons m-* s-'. As growth irradiance increased, several constituents of the "Mehler-peroxidase" pathway also increased: superoxide dismutase, ascorbate peroxidase, glutathione reductase, ascorbate, and glutathione. This occurred concomitantly with increases i n the xanthophyll cycle pool size and in the rate of nonphotochemical energy dissipation under steady-state conditions. There was no evidence for photosystem II overreduction in plants grown at high irradiance, although the reduction state of the stromal NADP pool, estimated from measurements of NADP-malate dehydrogenase activity, was greater than 60% in V. major and S. arboricola. Ascorbate, which removes reactive O, species generated by O, photoreduction in the chloroplast and serves as a reductant for the conversion of the xanthophyll cycle pigments to the de-epoxidized forms A plus 2, generally exhibited the most dramatic increases in response to growth irradiance. We conclude from these results that O, photoreduction occurs at higher rates i n leaves acclimated to high irradiance, despite increases in xanthophyll cycle-dependent energy dissipation, and that increases in leaf antioxidants protect against this potential oxidative stress.Plants acclimated to high irradiance use various mechanisms to protect the photosynthetic apparatus against the deleterious effects of excess light absorption. Much attention has been focused on elucidating the role of the xanthophyll cycle in the dissipation of excess excitation energy in the light-harvesting antennae . The associated increases in the xanthophyll cycle pool size, V plus A plus Z, and the capacity for nonphotochemical energy dissipation, NPQ, is a well-characterized and apparently fundamental acclimatory response to high PFDs (Bjorkman and Demmig-Adams, 1994; DemmigAdams and Adams, 1994). Xanthophyll cycle-dependent energy dissipation lowers the photon efficiency of PSII, thus providing a mechanism to balance the synthesis of ATP and NADPH with the rate at which these metabolites can be utilized in photosynthesis (Foyer, 1993 , 1995) or serve a regulatory role by augmenting the transthylakoid proton gradient (Schreiber and Neubauer, 1990). Although the magnitude of O,-dependent flux in vivo remains controversial, direct measurements using MS suggest that in C, plants up to 25% of the total noncyclic electron transport is consumed by this process at light saturation (Canvin et al., 1980;Badger, 1985;Osmond and Grace, 1995). Protection against the possible toxicity of ROS is afforded by an integrated system of enzymatic and nonenzymatic antioxidants that are concentrated in the chloroplast (Asada, 1994). The superoxide anion, the initial product of photosynthetic O, reduction, is either dismuted by SOD or reduced by ascorbate to H,O,, which is t...