Barley leaves were exposed for several min to a white light of photon flux density 1000 micromol m-2 s-1, leading to a massive conversion of the xanthophyll violaxanthin to antheraxanthin and zeaxanthin in the absence of lipid peroxidation. Using electron spin resonance spectroscopy and different spin-labeled stearate probes, we observed that this light treatment noticeably decreased thylakoid membrane lipid fluidity. The light-induced membrane rigidification (i) was proportional to the amount of zeaxanthin present in the membranes, (ii) was blocked by dithiothreitol, a potent inhibitor of the violaxanthin de-epoxidase, (iii) was slowly reversible in the dark, (iv) was not observed in thylakoids of an Arabidopsis mutant that has no xanthophyll cycle and (v) was accompanied by a substantial increase in the thermostability of the ionic permeability properties of the thylakoid membranes. The amount of xanthophyll-cycle pigments found in photosystem II was observed to significantly decrease after illumination. Photoacoustic and chlorophyll fluorometric analyses of the illuminated leaves revealed that strong illumination decreased the quantum yield of photosynthetic oxygen evolution and the pigment antenna size of photosystem II in green light (preferentially absorbed by carotenoids) but not in red light (absorbed by chlorophylls only). Taken together in the light of previous in vitro data on carotenoids incorporated into artificial membranes, our results indicate that the xanthophyll cycle could be an 'emergency mechanism' that rapidly provides thylakoid membrane lipids with rigidifying carotenoid molecules upon sudden increase in light intensity. The significance of this mechanism for the membrane function and adaptation to stressful light and temperature conditions is discussed.