Bicarbonate protects the water-oxidizing complex of photosystem II against thermoinactivation in intact Chlamydomonas reinhardtii cells

Abstract

Currently available data about bicarbonate (BC) action on the Mn-containing water-oxidizing complex (WOC) of the photosystem II (PSII) were obtained almost solely in vitro, e.g. on subchloroplast membrane fragments enriched with PSII. To investigate the in vivo BC effect on the PSII donor side, we used the method of dark thermoinactivation of intact Chlamydomonas reinhardtii cells. Photosynthetic activity of PSII was measured as photoinduced changes in the PSII chlorophyll fluorescence yield and as the rate of photosynthetic oxygen evolution. To exclude a “direct” effect of the absence of BC on the PSII activity, before measurements of the photosynthetic activity, the concentration of BC in all samples was equalized by addition of NaHCO3 to each of them (except for those that contained 5 mM of NaHCO3 during thermoinactivation) to reach the final concentration of 5 mM. This allowed registering only so-called “irreversible” (i.e., not reversible by subsequent addition of BC) effect of the absence of BC during thermoinactivation. It was shown that, if 5 mM NaHCO3 was added to the medium before thermoinactivation, the rate of inactivation of the PSII donor side was lower than in BC-depleted medium 1.5-to 2-fold. The obtained results are interpreted as an indication that BC protects the donor side of PSII against thermoinactivation in vivo, in intact C. reinhardtii cells. This proves the correctness of the earlier proposition that BC is an integral constituent of the Mn-containing water-oxidizing complex of PSII.