Genes encoding A-type flavoproteins are essential for photoreduction of O2 in cyanobacteria.

O(2) photoreduction by photosynthetic electron transfer, the Mehler reaction, was observed in all groups of oxygenic photosynthetic organisms, but the electron transport chain mediating this reaction remains unidentified. We provide the first evidence for the involvement of A-type flavoproteins that reduce O(2) directly to water in vitro. Synechocystis sp. strain PCC 6803 mutants defective in flv1 and flv3, encoding A-type flavoproteins, failed to exhibit O(2) photoreduction but performed normal photosynthesis and respiration. We show that the light-enhanced O(2) uptake was not due to respiration or photorespiration. After dark acclimation, photooxidation of P(700) was severely depressed in mutants Deltaflv1 and Deltaflv3 but recovered after light activation of CO(2) fixation, which gives P(700) an additional electron acceptor. Inhibition of CO(2) fixation prevented recovery but scarcely affected P(700) oxidation in the wild-type, where the Mehler reaction provides an alternative route for electrons. We conclude that the source of electrons for O(2) photoreduction is PSI and that the highly conserved A-type flavoproteins Flv1 and Flv3 are essential for this process in vivo. We propose that in cyanobacteria, contrary to eukaryotes, the Mehler reaction produces no reactive oxygen species and may be evolutionarily related to the response of anaerobic bacteria to O(2).

Massive light-dependent cycling of inorganic carbon between oxygenic photosynthetic microorganisms and their surroundings.

Membrane inlet mass spectrometry indicated massive light-dependent cycling of inorganic carbon between the medium and the cells of various phytoplankton species representing the main groups of aquatic primary producers. These included diatoms, symbiotic and free living dinoflagellates, a coccolithophorid, a green alga and filamentous and single cell cyanobacteria. These organisms could maintain an ambient CO(2) concentration substantially above or below that expected at chemical equilibrium with HCO(3) (-). The coccolithophorid Emiliania huxleyi shifted from net CO(2) uptake to net CO(2) efflux with rising light intensity. Differing responses of CO(2) uptake and CO(2) fixation to changing light intensity supported the notion that these two processes are not compulsorily linked. Simultaneous measurements of CO(2) and O(2) exchange and of the fluorescence parameters in Synechococcus sp. strain PCC 7942, showed that CO(2) uptake can serve as a sensitive probe of the energy status of the photosynthetic reaction centers. However, during transitions in light intensity, changes in CO(2) uptake did not accord with those expected from fluorescence change. Quantification of the net fluxes of CO(2), HCO(3) (-) and of photosynthesis at steady-state revealed that substantial HCO(3) (-) efflux accompanied CO(2) uptake and fixation in the case of 'CO(2) users'. On the other hand, 'HCO(3) (-) users' were characterized by a rate of net CO(2) uptake below that of CO(2) fixation. The results support the notion that entities associated with the CCM function not only in raising the CO(2) concentration at the site of Rubisco; they may also serve as a means of diminishing photodynamic damage by dissipating excess light energy.

CO2 CONCENTRATING MECHANISMS IN PHOTOSYNTHETIC MICROORGANISMS.

Many microorganisms possess inducible mechanisms that concentrate CO2 at the carboxylation site, compensating for the relatively low affinity of Rubisco for its substrate, and allowing acclimation to a wide range of CO2 concentrations. The organization of the carboxysomes in prokaryotes and of the pyrenoids in eukaryotes, and the presence of membrane mechanisms for inorganic carbon (Ci) transport, are central to the concentrating mechanism. The presence of multiple Ci transporting systems in cyanobacteria has been indicated. Certain genes involved in structural organization, Ci transport and the energization of the latter have been identified. Massive Ci fluxes associated with the CO2-concentrating mechanism have wide-reaching ecological and geochemical implications.