caa a22 4500 445889470 CHVBK 20180317145611.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s11120-011-9634-4 doi (NATIONALLICENCE)springer-10.1007/s11120-011-9634-4 Localization of putative carbonic anhydrases in two marine diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana [Elektronische Daten] [Masaaki Tachibana, Andrew Allen, Sae Kikutani, Yuri Endo, Chris Bowler, Yusuke Matsuda] It is believed that intracellular carbonic anhydrases (CAs) are essential components of carbon concentrating mechanisms in microalgae. In this study, putative CA-encoding genes were identified in the genome sequences of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. Subsequently, the subcellular localizations of the encoded proteins were determined. Nine and thirteen CA sequences were found in the genomes of P. tricornutum and T. pseudonana, respectively. Two of the β-CA genes in P. tricornutum corresponded to ptca1 and ptca2 identified previously. Immunostaining transmission electron microscopy of a PtCA1:YFP fusion expressed in the cells of P. tricornutum clearly showed the localization of PtCA1 within the central part of the pyrenoid structure in the chloroplast. Besides these two β-CA genes, P. tricornutum likely contains five α- and two γ-CA genes, whereas T. pseudonana has three α-, five γ-, four δ-, and one ζ-CA genes. Semi-quantitative reverse transcription PCR performed on mRNA from the two diatoms grown in changing light and CO2 conditions revealed that levels of six putative α- and γ-CA mRNAs in P. tricornutum did not change between cells grown in air-level CO2 and 5% CO2. However, mRNA levels of one putative α-CA gene, CA-VII in P. tricornutum, were reduced in the dark compared to that in the light. In T. pseudonana, mRNA accumulation levels of putative α-CA (CA-1), ζ-CA (CA-3) and δ-CA (CA-7) were analyzed and all levels found to be significantly reduced when cells were grown in 0.16% CO2. Intercellular localizations of eight putative CAs were analyzed by expressing GFP fusion in P. tricornutum and T. pseudonana. In P. tricornutum, CA-I and II localized in the periplastidial compartment, CA-III, VI, VII were found in the chloroplast endoplasmic reticulum, and CA-VIII was localized in the mitochondria. On the other hand, T. pseudonana CA-1 localized in the stroma and CA-3 was found in the periplasm. These results suggest that CAs are constitutively present in the four chloroplastic membrane systems in P. tricornutum and that CO2 responsive CAs occur in the pyrenoid of P. tricornutum, and in the stroma and periplasm of T. pseudonana. Springer Science+Business Media B.V., 2011 Marine diatom nationallicence Carbonic anhydrase nationallicence Inorganic carbon concentrating mechanism nationallicence Pyrenoid nationallicence Rubisco : Ribulose-1,5-bisphosphate carboxylase/oxygenase nationallicence CA : Carbonic anhydrase nationallicence CER : Chloroplast endoplasmic reticulum nationallicence PPC : Periplastidial compartment nationallicence BLS : Blob-like structure nationallicence CE : Chloroplast envelope nationallicence CCM : Inorganic carbon concentrating mechanism nationallicence TEM : Transmission electron microscopy nationallicence Tachibana Masaaki Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut Allen Andrew Environmental and Evolutionary Genomics Section, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), CNRS UMR8197 INSERM U1024, Ecole Normale Supérieure, 46 rue d'Ulm, 75005, Paris, France aut Kikutani Sae Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut Endo Yuri Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut Bowler Chris Environmental and Evolutionary Genomics Section, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), CNRS UMR8197 INSERM U1024, Ecole Normale Supérieure, 46 rue d'Ulm, 75005, Paris, France aut Matsuda Yusuke Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut Photosynthesis Research Springer Netherlands 109/1-3(2011-09-01), 205-221 0166-8595 109:1-3<205 2011 109 11120 https://doi.org/10.1007/s11120-011-9634-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11120-011-9634-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tachibana Masaaki Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut NATIONALLICENCE 700 1- Allen Andrew Environmental and Evolutionary Genomics Section, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), CNRS UMR8197 INSERM U1024, Ecole Normale Supérieure, 46 rue d'Ulm, 75005, Paris, France aut NATIONALLICENCE 700 1- Kikutani Sae Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut NATIONALLICENCE 700 1- Endo Yuri Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut NATIONALLICENCE 700 1- Bowler Chris Environmental and Evolutionary Genomics Section, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), CNRS UMR8197 INSERM U1024, Ecole Normale Supérieure, 46 rue d'Ulm, 75005, Paris, France aut NATIONALLICENCE 700 1- Matsuda Yusuke Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, 669-1337, Sanda, Hyogo, Japan aut NATIONALLICENCE 773 0- Photosynthesis Research Springer Netherlands 109/1-3(2011-09-01), 205-221 0166-8595 109:1-3<205 2011 109 11120 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer