title: The carbon concentrating mechanism in Chlamydomonas reinhardtii: finding the missing pieces.
abstract: The photosynthetic, unicellular green alga, Chlamydomonas reinhardtii, lives in environments that often contain low concentrations of CO2 and HCO3 (-), the utilizable forms of inorganic carbon (Ci). C. reinhardtii possesses a carbon concentrating mechanism (CCM) which can provide suitable amounts of Ci for growth and development. This CCM is induced when the CO2 concentration is at air levels or lower and is comprised of a set of proteins that allow the efficient uptake of Ci into the cell as well as its directed transport to the site where Rubisco fixes CO2 into biomolecules. While several components of the CCM have been identified in recent years, the picture is still far from complete. To further improve our knowledge of the CCM, we undertook a mutagenesis project where an antibiotic resistance cassette was randomly inserted into the C. reinhardtii genome resulting in the generation of 22,000 mutants. The mutant collection was screened using both a published PCR-based approach (Gonzalez-Ballester et al. 2011) and a phenotypic growth screen. The PCR-based screen did not rely on a colony having an altered growth phenotype and was used to identify colonies with disruptions in genes previously identified as being associated with the CCM-related gene. Eleven independent insertional mutations were identified in eight different genes showing the usefulness of this approach in generating mutations in CCM-related genes of interest as well as identifying new CCM components. Further improvements of this method are also discussed.
authors.affiliation: Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, 70803, USA.
authors.forename: Nadine
authors.initials: N
authors.lastname: Jungnick
authors.forename: Yunbing
authors.initials: Y
authors.lastname: Ma
authors.forename: Bratati
authors.initials: B
authors.lastname: Mukherjee
authors.forename: Julie C
authors.initials: JC
authors.lastname: Cronan
authors.forename: Dequantarius J
authors.initials: DJ
authors.lastname: Speed
authors.forename: Susan M
authors.initials: SM
authors.lastname: Laborde
authors.forename: David J
authors.initials: DJ
authors.lastname: Longstreth
authors.forename: James V
authors.initials: JV
authors.lastname: Moroney
mesh_terms: D002244:Carbon / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D018745:Genome, Plant / Q000235:genetics
chemical_list: D002244:Carbon
references.citation: EMBO J. 1998 Aug 10;17(5):1208-16
references.pmid: 9482718
references.citation: Plant Cell. 2004 Jun;16(6):1466-77
references.pmid: 15155888
references.citation: Annu Rev Plant Biol. 2005;56:99-131
references.pmid: 15862091
references.citation: Plant J. 2002 May;30(3):301-13
references.pmid: 12000678
references.citation: Plant Physiol. 2011 Jun;156(2):884-96
references.pmid: 21527423
references.citation: Am J Physiol Cell Physiol. 2008 Jun;294(6):C1371-7
references.pmid: 18400985
references.citation: Plant Physiol. 2011 Jan;155(1):56-63
references.pmid: 20940348
references.citation: Plant Physiol. 1983 Oct;73(2):273-6
references.pmid: 16663207
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):18228-33
references.pmid: 15596724
references.citation: Plant Physiol. 2005 Feb;137(2):545-56
references.pmid: 15653810
references.citation: Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5990-5
references.pmid: 19321421
references.citation: Photosynth Res. 2011 Sep;109(1-3):133-49
references.pmid: 21365258
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 27;103(26):10110-5
references.pmid: 16777959
references.citation: Plant Physiol. 1985 Sep;79(1):177-83
references.pmid: 16664365
references.citation: Plant Cell. 2012 May;24(5):1860-75
references.pmid: 22634764
references.citation: Plant Physiol. 1987 Mar;83(3):460-3
references.pmid: 16665269
references.citation: J Exp Bot. 2013 Jan;64(3):753-68
references.pmid: 23028015
references.citation: Physiol Plant. 2008 May;133(1):15-26
references.pmid: 18405332
references.citation: J Exp Bot. 2013 Jan;64(3):769-86
references.pmid: 23345319
references.citation: Science. 2003 Mar 7;299(5612):1572-5
references.pmid: 12624266
references.citation: Plant Mol Biol. 2004 Sep;56(1):125-32
references.pmid: 15604732
references.citation: Plant Cell Physiol. 2010 Sep;51(9):1453-68
references.pmid: 20660228
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5341-6
references.pmid: 11309511
references.citation: Photosynth Res. 2011 Sep;109(1-3):47-57
references.pmid: 21359551
references.citation: Plant Physiol. 1999 Jul;120(3):757-64
references.pmid: 10398710
references.citation: Gene. 2001 Oct 17;277(1-2):221-9
references.pmid: 11602359
references.citation: Plant Cell. 2012 May;24(5):1876-93
references.pmid: 22634760
references.citation: Plant Physiol. 1989 Apr;89(4):1213-9
references.pmid: 16666686
references.citation: Plant Physiol. 1993 Apr;101(4):1195-1199
references.pmid: 12231773
references.citation: Plant Methods. 2011 Jul 27;7:24
references.pmid: 21794168
references.citation: Plant Physiol. 2011 Jan;155(1):20-6
references.pmid: 20923885
references.citation: Plant Physiol. 1998 Apr;116(4):1585-91
references.pmid: 9536077
references.citation: Plant Physiol. 2003 Dec;133(4):1854-61
references.pmid: 14605215
references.citation: Proc Natl Acad Sci U S A. 2012 Nov 20;109(47):19474-9
references.pmid: 23112177
references.citation: Curr Opin Biotechnol. 2008 Apr;19(2):153-9
references.pmid: 18374559
references.citation: Biochem J. 1997 Oct 1;327 ( Pt 1):51-7
references.pmid: 9355734
references.citation: Plant Physiol. 2004 Jul;135(3):1595-607
references.pmid: 15235119
references.citation: Philos Trans R Soc Lond B Biol Sci. 2012 Feb 19;367(1588):493-507
references.pmid: 22232762
references.citation: J Basic Microbiol. 2009 Feb;49(1):42-51
references.pmid: 19253331
references.citation: J Biol Chem. 2010 Jun 25;285(26):19833-41
references.pmid: 20424165
references.citation: Plant Mol Biol. 1996 May;31(2):443-8
references.pmid: 8756610
references.citation: Plant Physiol. 1991 Dec;97(4):1576-80
references.pmid: 16668587
references.citation: Eukaryot Cell. 2007 Aug;6(8):1251-9
references.pmid: 17557885
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5347-52
references.pmid: 11287669
references.citation: Photosynth Res. 2013 Nov;117(1-3):121-31
references.pmid: 23771683
references.citation: Plant Cell. 2010 Sep;22(9):3105-17
references.pmid: 20870960
references.citation: Plant Physiol. 2003 Aug;132(4):2267-75
references.pmid: 12913181
references.citation: Plant Physiol. 1989 Mar;89(3):897-903
references.pmid: 16666639
references.citation: Biochim Biophys Acta. 2012 Aug;1817(8):1248-55
references.pmid: 22709623
references.citation: Science. 2012 Jun 29;336(6089):1671-2
references.pmid: 22745421
references.citation: PLoS One. 2012;7(11):e49063
references.pmid: 23139834
references.citation: Plant Physiol. 2005 Dec;139(4):1959-69
references.pmid: 16306144
references.citation: Annu Rev Plant Biol. 2010;61:235-61
references.pmid: 20192734
references.citation: Photosynth Res. 2011 Sep;109(1-3):281-96
references.pmid: 21327536
references.citation: Protist. 2006 Oct;157(4):421-33
references.pmid: 16905358
references.citation: Plant Physiol. 1983 Oct;73(2):268-72
references.pmid: 16663206
references.citation: Genetics. 1998 Apr;148(4):1821-8
references.pmid: 9560396