Nonphotochemical quenching of excitation energy in photosystem II.: A picosecond time-resolved study of the low yield of chlorophyll a fluorescence induced by single-turnover flash in isolated spinach thylakoids

Chlorophyll a fluorescence emission is widely used as a noninvasive measure of a number of parameters related to photosynthetic efficiency in oxygenic photosynthetic organisms. The most important component for the estimation of photochemistry is the relative increase in fluorescence yield between dark-adapted samples which have a maximal capacity for photochemistry and a minimal fluorescence yield (F-o) and light-saturated samples where photochemistry is saturated and fluorescence yield is maximal (F-m). However, when photosynthesis is saturated with a short (less than 50 mu s) flash of light, which induces only one photochemical turnover of photosystem II, the maximal fluorescence yield is significantly lower (F-sat) than when saturation is achieved with a millisecond duration multiturnover flash (F-m). To investigate the origins of the difference in fluorescence yield between these two conditions, our time resolved fluorescence apparatus was modified to allow collection of picosecond time-resolved decay kinetics over a short time window immediately following a saturating single-turnover flash (F-sat) as well as after a multiturnover saturating pulse (F-m), Our data were analyzed with a global kinetic model based on an exciton radical pair equilibrium model for photosystem II, The difference between F-m and F-sat was modeled well by changing only the rate constant for quenching of excitation energy in the antenna of photosystem II. An antenna-based origin for the quenching was verified experimentally by the observation that addition of the antenna quencher 5-hydroxy-1,4-naphthoquinone to thylakoids under F-m conditions resulted in decay kinetics and modeled kinetic parameters very similar to those observed under F-sat conditions in the absence of added quinone. Our data strongly support the origin of low fluorescence yield at F-sat to be an antenna-based nonphotochemical quenching of excitation energy in photosystem II which has not usually been considered explicitly in calculations of photochemical and nonphotochemical quenching parameters. The implications of our data with respect to kinetic models for the excited-state dynamics of photosystem II and the practical applications of the fluorescence yield parameters F-m and F-sat to calculations of photochemical yield are discussed.