CO2-concentrating mechanisms of the potentially toxic dinoflagellate Protoceratium reticulatum (Dinophyceae, Gonyaulacales)

The low CO2 concentration in seawater poses severe restrictions on photosynthesis, especially on those species with form II RUBISCO. We found that the potentially toxic dinoflagellate Protoceratium reticulatum Clap. et J. Lachm. possesses a form II RUBISCO. To cast some light on the mechanisms this organism undergoes to cope with low CO2 availability, we compared cells grown at atmospheric (370 ppm) and high (5000 ppm) CO2 concentrations, with respect to a number of physiological parameters related to dissolved inorganic carbon (DIC) acquisition and assimilation. The photosynthetic affinity for DIC was about one order of magnitude lower in cells cultivated at high [CO2]. Endpoint pH-drift experiments suggest that P. reticulatum was not able to efficiently use HCO 3 under our growth conditions. Only internal carbonic anhydrase (CA) activity was detected, and its activity decreased by about 60% in cells cultured at high [CO2]. Antibodies raised against a variety of algal CAs were used for Western blot analysis: P. reticulatum extracts only cross-reacted with anti-beta-CA sera, and the amount of immunoreactive protein decreased in cells grown at high [CO2]. No pyrenoids were observed under all growth conditions. Our data indicate that P. reticulatum has an inducible carbon-concentrating mechanism (CCM) that operates in the absence of pyrenoids and with little intracellular CO2 accumulation. Calculations on the impact of the CA activity to photosynthetic growth [CO2] suggest that it is an essential component of the CCM of P. reticulatum and is necessary to sustain the photosynthetic rates observed at ambient CO2.