title: The Chlamydomonas reinhardtii cia3 mutant lacking a thylakoid lumen-localized carbonic anhydrase is limited by CO2 supply to rubisco and not photosystem II function in vivo.
abstract: The Chlamydomonas reinhardtii cia3 mutant has a phenotype indicating that it requires high-CO(2) levels for effective photosynthesis and growth. It was initially proposed that this mutant was defective in a carbonic anhydrase (CA) that was a key component of the photosynthetic CO(2)-concentrating mechanism (CCM). However, more recent identification of the genetic lesion as a defect in a lumenal CA associated with photosystem II (PSII) has raised questions about the role of this CA in either the CCM or PSII function. To resolve the role of this lumenal CA, we re-examined the physiology of the cia3 mutant. We confirmed and extended previous gas exchange analyses by using membrane-inlet mass spectrometry to monitor(16)O(2),(18)O(2), and CO(2) fluxes in vivo. The results demonstrate that PSII electron transport is not limited in the cia3 mutant at low inorganic carbon (Ci). We also measured metabolite pools sizes and showed that the RuBP pool does not fall to abnormally low levels at low Ci as might be expected by a photosynthetic electron transport or ATP generation limitation. Overall, the results demonstrate that under low Ci conditions, the mutant lacks the ability to supply Rubisco with adequate CO(2) for effective CO(2) fixation and is not limited directly by any aspect of PSII function. We conclude that the thylakoid CA is primarily required for the proper functioning of the CCM at low Ci by providing an ample supply of CO(2) for Rubisco.
authors.affiliation: University of New Mexico, Department of Biology, Albuquerque, New Mexico 87131, USA.
authors.forename: David Thomas
authors.initials: DT
authors.lastname: Hanson
authors.forename: Linda A
authors.initials: LA
authors.lastname: Franklin
authors.forename: Goran
authors.initials: G
authors.lastname: Samuelsson
authors.forename: Murray R
authors.initials: MR
authors.lastname: Badger
mesh_terms: D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects; D008027:Light; D009154:Mutation / Q000235:genetics*; D010100:Oxygen / Q000378:metabolism; D010788:Photosynthesis / Q000528:radiation effects; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism*; D020524:Thylakoids / Q000201:enzymology*
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D002245:Carbon Dioxide; D012273:Ribulose-Bisphosphate Carboxylase; D002256:Carbonic Anhydrases; D010100:Oxygen