Limited sensitivity of pigment photo-oxidation in isolated thylakoids to singlet excited state quenching in photosystem II antenna

Light-induced pigment oxidation and its relation to excited state quenching in photosystems antennae have been investigated in isolated thylakoids. The results indicate that (i) chlorophyll oxidation takes place in two sequential steps. A slow initial phase is followed by a steep increase in the bleaching rate when more than one quarter of the chromophores are oxidised. (ii) During the initial slow phase, the carotenoid pool is bleached with an apparent rate which is about three times faster than that found for chlorophyll a and more than six times faster than that of chlorophyll b. (iii) Pigment bleaching has been observed both in photosystem I and photosystem II, and it has been possible to estimate a similar carotenoid bleaching rate in the two photosystems. (iv) The protection conferred by singlet state quenchers in the initial slow phase of pigment oxidation is modest. Taking into consideration that both the photosystems are subjected to the oxidative treatment, a somewhat larger protective effect than those estimated for photo-inhibition in thylakoids [S. Santabarbara, F.M. Garlaschi, G. Zucchelli, R.C. Jennings, Biochim. Biophys. Acta 1409 (1999) 165-170] can be computed, although it is less than 50% of the expected level on the basis of the observed reciprocity to the number of incident photons. (v) Pigment oxidation is associated with the loss of membrane ultra-structure, which is interpreted as originating from a decrease in grana stacking. The dynamics of loss of membrane ultra-structure parallel the phases observed for chlorophyll photo-bleaching.