The physiology and genetics of CO2 concentrating mechanisms in model diatoms.

Diatoms, a diverse and ecologically-important group of unicellular algae, use a CO2 concentrating mechanism to enhance the performance of RubisCO and overcome the limited availability of CO2 in their habitats. The recent development of genetic manipulation techniques for the model diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana and the sequencing of their genomes have enabled the rapid identification of genes involved in their CO2 concentrating mechanisms (CCMs). These include numerous carbonic anhydrases (CAs), which are localized to distinct subcellular compartments in the two diatom species, and putative bicarbonate transporters, one of which has been functionally characterized. New physiological data on the P. tricornutum CCM are consistent with this molecular data and suggest that the major driver of the CCM is a 'chloroplast-pump' that actively transports bicarbonate into the chloroplast. In T. pseudonana, the localization of a CA in the chloroplast stroma presents a paradox as this would be expected to impede function of a biophysical CCM, though the recent proposal of a modified C4 CCM offers a potential explanation.

Hopkinson BM ，Dupont CL ，Matsuda Y 《-》

Localization of putative carbonic anhydrases in two marine diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana.

Tachibana M ，Allen AE ，Kikutani S ，Endo Y ，Bowler C ，Matsuda Y ... -  《-》

Molecular aspects of the biophysical CO2-concentrating mechanism and its regulation in marine diatoms.

Diatoms operate a CO2-concentrating mechanism (CCM) that drives upwards of 20% of annual global primary production. Recent progress in CCM research in the marine pennate diatom Phaeodactylum tricornutum revealed that this diatom directly takes up HCO3- from seawater through low-CO2-inducible plasma membrane HCO3- transporters, which belong to the solute carrier (SLC) 4 family. Apart from this, studies of carbonic anhydrases (CAs) in diatoms have revealed considerable diversity in classes and localization among species. This strongly suggests that the CA systems, which control permeability and flux of dissolved inorganic carbon (DIC) by catalysing reversible CO2 hydration, have evolved from diverse origins. Of particular interest is the occurrence of low-CO2-inducible external CAs in the centric marine diatom Thalassiosira pseudonana, offering a strategy of CA-catalysed initial CO2 entry via passive diffusion, contrasting with active DIC transport in P. tricornutum. Molecular mechanisms to transport DIC across chloroplast envelopes are likely also through specific HCO3- transporters, although details have yet to be elucidated. Furthermore, recent discovery of a luminal θ-CA in the diatom thylakoid implied a common strategy in the mechanism to supply CO2 to RubisCO in the pyrenoid, which is conserved among green algae and some heterokontophytes. These results strongly suggest an occurrence of convergent coevolution between the pyrenoid and thylakoid membrane in aquatic photosynthesis.

Tsuji Y ，Nakajima K ，Matsuda Y 《-》

Diversity of CO2-concentrating mechanisms and responses to CO2 concentration in marine and freshwater diatoms.

The presence of CO2-concentrating mechanisms (CCMs) is believed to be one of the characteristics that allows diatoms to thrive in many environments and to be major contributors to global productivity. Here, the type of CCM and the responses to variable CO2 concentration were studied in marine and freshwater diatoms. At 400 ppm, there was a large diversity in physiological and biochemical mechanisms among the species. While Phaeodactylum tricornutum mainly used HCO3-, Thalassiosira pseudonana mainly used CO2. Carbonic anhydrase was an important component of the CCM in all species and C4 metabolism was absent, even with T. weissflogii. For all species, at 20 000 ppm, the affinity for dissolved inorganic carbon was lower than at 400 ppm CO2 and the reliance on CO2 was higher. Despite the difference in availability of inorganic carbon in marine and fresh waters, there were only small differences in CCMs between species from the two environments, and Navicula pelliculosa behaved similarly when grown in the two environments. The results suggest that species-specific differences are great, and more important than environmental differences in determining the nature and effectiveness of the CCM in diatoms.

Clement R ，Jensen E ，Prioretti L ，Maberly SC ，Gontero B ... -  《-》

The nature of the CO2 -concentrating mechanisms in a marine diatom, Thalassiosira pseudonana.

Diatoms are widespread in aquatic ecosystems where they may be limited by the supply of inorganic carbon. Their carbon dioxide-concentrating mechanisms (CCMs) involving transporters and carbonic anhydrases (CAs) are well known, but the contribution of a biochemical CCM involving C4 metabolism is contentious. The CCM(s) present in the marine-centric diatom, Thalassiosira pseudonana, were studied in cells exposed to high or low concentrations of CO2 , using a range of approaches. At low CO2 , cells possessed a CCM based on active uptake of CO2 (70% contribution) and bicarbonate, while at high CO2 , cells were restricted to CO2 . CA was highly and rapidly activated on transfer to low CO2 and played a key role because inhibition of external CA produced uptake kinetics similar to cells grown at high CO2 . The activities of phosphoenolpyruvate (PEP) carboxylase (PEPC) and the PEP-regenerating enzyme, pyruvate phosphate dikinase (PPDK), were lower in cells grown at low than at high CO2 . The ratios of PEPC and PPDK to ribulose bisphosphate carboxylase were substantially lower than 1, even at low CO2 . Our data suggest that the kinetic properties of this species results from a biophysical CCM and not from C4 type metabolism.

Clement R ，Dimnet L ，Maberly SC ，Gontero B ... -  《-》