The influence of zeaxanthin on high-energy-state quenching (q(E)) of room temperature chlorophyll fluorescence and on the quantum yield of oxygen evolution(Phi(O2)) has been studied with isolated spinach thylakoids. A set of three different pretreatments was tested which all led to thylakoids with high contents of zeaxanthin. Depending on the presence of light the pretreatments differed strongly with respect to their effect on nonphotochemical fluorescence quenching. Independently from the extent of changes in nonphotochemical energy dissipation in the membrane energized state as reflected by nonphotochemical quenching the light pretreatments caused also changes in nonphotochemical energy dissipation in the nonenergized state as measured by the quantum yield of oxygen evolution (Phi(O2)) in uncoupled thylakoids. Zeaxanthin was not involved in the decrease of (Phi(O2)).