A mutant of Chlamydomonas reinhardtii that cannot acclimate to low CO2 conditions has an insertion in the Hdh1 gene
James E. Adams A , Sergio L. Colombo A , Catherine B. Mason A , Ruby A. Ynalvez A , Baran Tural A and James V. Moroney A BA Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.
B Corresponding author. Email: btmoro@lsu.edu
Functional Plant Biology 32(1) 55-66 https://doi.org/10.1071/FP04119
Submitted: 6 July 2004 Accepted: 5 November 2004 Published: 21 January 2005
Abstract
Photosynthetic microorganisms must acclimate to environmental conditions, such as low CO2 environments or high light intensities, which may lead to photo-oxidative stress. In an effort to understand how photosynthetic microorganisms acclimate to these conditions, Chlamydomonas reinhardtii was transformed using the BleR cassette, selected for Zeocin resistance and screened for colonies that showed poor growth at low CO2 levels. One of the insertional mutants obtained, named slc-230, was shown to have a BleR insert in the first exon of Hdh1, a novel, single copy gene. The predicted Hdh1 gene product has similarity to bacterial haloacid dehalogenase-like proteins, a protein family that includes phosphatases and epoxide hydrolases. In addition, Hdh1 is predicted to be localised to the chloroplast or mitochondria in C. reinhardtii. It was found that a genomic copy of wild type Hdh1 can complement slc-230.
Physiological studies were conducted to determine the effects of the altered expression of Hdh1 in slc-230. slc-230 grows slowly autotrophically in low CO2, exhibits a lower affinity for inorganic carbon, a decreasing photosynthetic rate over time and a lower content of chlorophylls and quenching xanthophylls than wild type cells. Some possible roles of Hdh1 in the acclimation to low CO2 conditions are discussed.
Keywords: Chlamydomonas reinhardtii, growth on low CO2, Hdh1, haloacid dehalogenase-like gene, insertional mutagenesis.
Altschul SF,
Madden TL,
Schäffer AA,
Zhang J,
Zhang Z,
Miller W, Lipman DJ
(1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25, 3389–3402.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Arnon DI
(1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris.
Plant Physiology 24, 1–15.
Asamizu E,
Miura K,
Kucho K,
Inoue Y,
Fukuzawa H,
Ohyama K,
Nakamura Y, Tabata S
(2000) Generation of expressed sequence tags from low-CO2 and high-CO2 adapted cells of Chlamydomonas reinhardtii.
DNA Research 7, 305–307.
| PubMed |
Baroli I,
Do AD,
Yamane T, Niyogi KK
(2003) Zeaxanthin accumulation in the absence of a functional xanthophyll cycle protects Chlamydomonas reinhardtii from photooxidative stress. The Plant Cell 15, 992–1008.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Baroli I,
Gutman BL,
Ledford HK,
Shin JW,
Chin BL,
Havaux M, Niyogi KK
(2004) Photo-oxidative stress in a xanthophyll-deficient mutant of Chlamydomonas.
Journal of Biological Chemistry 279, 6337–6344.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Borkhsenious ON,
Mason CB, Moroney JV
(1998) The intracellular localization of ribulose-1,5-bisphosphate carboxylase / oxygenase in Chlamydomonas reinhardtii.
Plant Physiology 116, 1585–1591.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Buffan-Dubau E, Carman KR
(2000) Diel feeding behavior of meiofauna and their relationships with microalgal resources. Limnology and Oceanography 45, 381–395.
Burow MD,
Chen Z-Y,
Mouton TM, Moroney JV
(1996) Isolation of cDNA clones induced upon transfer of Chlamydomonas reinhardtii cells to low CO2. Plant Molecular Biology 31, 443–448.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Colman B,
Huertas IE,
Bhatti S, Dason JS
(2002) The diversity of inorganic carbon mechanisms in eukaryotic algae. Functional Plant Biology 29, 261–270.
| Crossref | GoogleScholarGoogle Scholar |
Colombo SL,
Pollock SV,
Eger KA,
Godfrey AC,
Adams JE,
Mason CB, Moroney JV
(2002) Use of the bleomycin resistance gene to generate tagged insertional mutants of Chlamydomonas reinhardtii that require elevated CO2 for optimal growth. Functional Plant Biology 29, 231–241.
| Crossref | GoogleScholarGoogle Scholar |
Davies JP,
Yildiz FH, Grossman AR
(1996) Sac1, a putative regulator that is critical to survival of Chlamydomonas reinhardtii during sulfur deprivation. EMBO Journal 15, 2150–2159.
| PubMed |
Davies JP,
Yildiz FH, Grossman AR
(1999) Sac3, an Snf1-like serine / threonine kinase that positively and negatively regulates the responses of Chlamydomonas to sulfur limitation. The Plant Cell 11, 1179–1190.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Dietmaier W,
Fabry S,
Huber H, Schmitt R
(1995) Analysis of a family of ypt genes and their products from Chlamydomonas reinhardtii.
Gene 158, 41–50.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Emanuelsson O,
Nielsen H,
Brunak S, von Heijne G
(2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. Journal of Molecular Biology 300, 1005–1016.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Eriksson M,
Karlsson J,
Ramazanov Z,
Gardeström P, Samuelsson G
(1996) Discovery of an algal mitochondrial carbonic anhydrase: molecular cloning and characterization of a low-CO2-induced polypeptide in Chlamydomonas reinhardtii.
Proceedings of the National Academy of Sciences USA 93, 12 031–12 034.
| Crossref | GoogleScholarGoogle Scholar |
Ferris PJ,
Armbrust EV, Goodenough UW
(2002) Genetic structure of the mating-type locus of Chlamydomonas reinhardtii.
Genetics 160, 181–200.
| PubMed |
Fujiwara S,
Fukuzawa H,
Tachiki A, Miyachi S
(1990) Structure and differential expression of two genes encoding carbonic anhydrase in Chlamydomonas reinhardtii.
Proceedings of the National Academy of Sciences USA 87, 9779–9783.
Fukuzawa H,
Miura K,
Ishizaki K,
Kucho K,
Saito T,
Kohinata T, Ohyama K
(2001) Ccm1, a regulatory gene controlling the induction of a carbon-concentrating mechanism in Chlamydomonas reinhardtii by sensing CO2. Proceedings of the National Academy of Sciences USA 98, 5347–5352.
| Crossref | GoogleScholarGoogle Scholar |
Gussow D, Clackson T
(1989) Direct clone characterization from plaques and colonies by the polymerase chain reaction. Nucleic Acids Research 17, 4000–4000.
| PubMed |
Harris, EH (1989).
Im CS,
Zhang ZD,
Shrager J,
Chang CW, Grossman AR
(2003) Analysis of light and CO2 regulation in Chlamydomonas reinhardtii using genome-wide approaches. Photosynthesis Research 75, 111–125.
| Crossref | GoogleScholarGoogle Scholar |
Jeffrey, SW ,
Mantoura, RFC ,
and
Wright, SW (1997).
Kaplan A, Reinhold L
(1999) CO2 concentrating mechanisms in photosynthetic microorganisms. Annual Review of Plant Physiology and Plant Molecular Biology 50, 539–570.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Karlsson J,
Clarke AK,
Chen Z-Y,
Park Y-I,
Hugghins SY,
Husic HD,
Moroney JV, Samuelsson G
(1998) A novel α-type carbonic anhydrase associated with the thylakoid membrane in Chlamydomonas reinhardtii is required for growth at ambient CO2. EMBO Journal 17, 1208–1216.
| Crossref |
PubMed |
Kindle KL
(1990) High-frequency nuclear transformation of Chlamydomonas reinhardtii.
Proceedings of the National Academy of Sciences USA 87, 1228–1232.
Mitra M,
Lato SM,
Ynalvez RA,
Xiao Y, Moroney JV
(2004) Identification of a new chloroplast carbonic anhydrase in Chlamydomonas reinhardtii.
Plant Physiology 135, 173–182.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Miura K,
Yamano T,
Yoshioka S,
Kohinata T, Inoue Y , et al.
(2004) Expression profiling-based identification of CO2-responsive genes regulated by CCM1 controlling a carbon-concentrating mechanism in Chlamydomonas reinhardtii.
Plant Physiology 135, 1595–1607.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Moroney JV, Somanchi A
(1999) How do algae concentrate CO2 to increase the efficiency of photosynthetic carbon fixation? Plant Physiology 119, 9–16.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Niyogi KK,
Bjorkman O, Grossman AR
(1997)
Chlamydomonas xanthophyll cycle mutants identified by video imaging of chlorophyll fluorescence quenching. The Plant Cell 9, 1369–1380.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Pazour GJ,
Wilkerson CG, Witman GB
(1998) A dynein light chain is essential for the retrograde particle movement of intraflagellar transport (IFT). Journal of Cell Biology 141, 979–992.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Pollock SV,
Colombo SL,
Prout DL,
Godfrey AC, Moroney JV
(2003) Rubisco activase is required for optimal photosynthesis in the green alga Chlamydomonas reinhardtii in a low-CO2 atmosphere. Plant Physiology 133, 1854–1861.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Price GD,
Maeda S,
Omata T, Badger MR
(2002) Modes of active inorganic carbon uptake in the cyanobacterium, Synechococcus sp. PCC7942. Functional Plant Biology 29, 131–149.
| Crossref | GoogleScholarGoogle Scholar |
Raven JA
(1997) Inorganic carbon acquisition by marine autotrophs. Advances in Botanical Research 27, 85–209.
Rawat M, Moroney JV
(1995) Regulation of carbonic anhydrase and ribulose-1,5-bisphosphate carboxylase / oxygenase activase by light and CO2 in Chlamydomonas reinhardtii.
Plant Physiology 109, 937–944.
| PubMed |
Rodman JS, Wandinger-Ness A
(2000) RAB GTPases coordinate endocytosis. Journal of Cell Science 113, 183–192.
| PubMed |
Sambrook, J ,
Fritsch, EF ,
and
Maniatis, T (1989).
Schnell RA, Lefebvre PA
(1993) Isolation of the Chlamydomonas reinhardtii regulatory gene NIT2 by transposon tagging. Genetics 134, 737–747.
| PubMed |
Shimogawara K,
Fujiwara S,
Grossman A, Usuda H
(1998) High-efficiency transformation of Chlamydomonas reinhardtii by electroporation. Genetics 148, 1821–1828.
| PubMed |
Somanchi A,
Handley ER, Moroney JV
(1998)
Chlamydomonas reinhardtii cDNAs upregulated in low CO2 conditions: expression and analysis. Canadian Journal of Botany 76, 1003–1009.
| Crossref | GoogleScholarGoogle Scholar |
Sueoka N
(1960) Mitotic replication of deoxyribonucleic acids in Chlamydomonas reinhardtii.
Proceedings of the National Academy of Sciences USA 46, 83–91.
Wright SW,
Jeffrey SW,
Mantoura RFC,
Llewellyn CA,
Bjornland T,
Repeta D, Welschmeyer N
(1991) Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton. Marine Ecology Progress Series 77, 183–196.
