Skip to main content
SpringerLink
Log in

Molecular comparison of carbonic anhydrase from Flaveria species demonstrating different photosynthetic pathways

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

During the evolution of C4 plants from C3 plants, both the function and intracellular location of carbonic anhydrase (CA) have changed. To determine whether these changes are due to changes at the molecular level, we have studied the cDNA sequences and the expression of CA from Flaveria species demonstrating different photosynthetic pathways. In leaf extracts from F. bidentis (C4), F. brownii (C4-like), F. linearis (C3–C4) and F. pringlei (C3), two polypeptides of Mr 31 kDa and 35 kDa cross-reacted with anti-spinach CA antibodies. However, the relative labelling intensities of the two polypeptides differed depending on the species. Northern blot analysis indicated at least two CA transcripts are present in each Flaveria species with sizes ranging from 1.1 to 1.6 kb. Carbonic anhydrase cDNAs from all four Flaveria species studied encode an open reading frame for a polypeptide of 35–36 kDa. The amino acid sequences deduced from all four Flaveria cDNAs share at least 70% homology with the sequences of other dicot CAs. The F. bidentis (C4) CA sequence was found to be the least similar of the Flaveria proteins and, as most of the sequence dissimilarity was found in the first third of the CA molecule, these differences may be involved in the intracellular targeting of CA. A neighbour-joining tree inferred from CA amino acid sequences showed that the Flaveria CAs cluster with other dicot CAs forming a group distinct from those of monocot CAs and prokaryotic and Chlamydomonas periplasmic CAs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Atkins CA, Patterson BD, Graham D: Plant carbonic anhydrases. I. Distribution of types among species. Plant Physiol 50: 214–217 (1972).

    Google Scholar 

  2. Bracey MH, Christiansen J, Tovar P, Cramer SP, Bartlett SG: Spinach carbonic anhydrase: Investigation of the zinc-binding ligands by site-directed mutagenesis, elemental analysis, and EXAFS. Biochemistry 33: 13126–13131 (1994).

    Google Scholar 

  3. Brown WV, Smith BN: Grass evolution, the Kranz syndrome, 13C/12C ratios, and continental drift. Nature 239: 345–346 (1972).

    Google Scholar 

  4. Burnell JN, Hatch MD: Low bundle sheath carbonic anhydrase is apparently essential for effective C4 pathway operation. Plant Physiol 86: 1252–1256 (1988).

    Google Scholar 

  5. Burnell JN, Gibbs MJ, Mason JG: Spinach chloroplastic carbonic anhydrase: nucleotide sequence analysis of cDNA. Plant Physiol 92: 37–40 (1990).

    Google Scholar 

  6. Cavallaro A, Ludwig M, Burnell J: The nucleotide sequence of a complementary DNA encoding Flaveria bidentis carbonic anhydrase. FEBS Lett 350: 216–218 (1994).

    Google Scholar 

  7. Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159 (1987).

    Google Scholar 

  8. Dean C, Tamaki S, Dunsmuir P, Favreau M, Katayama C, Dooner H, Bedbrook J: mRNA transcripts of several plant genes are polyadenylated at multiple sites in vivo. Nucl Acids Res 14: 2229–2240 (1986).

    Google Scholar 

  9. Fawcett TW, Browse JA, Volokita M, Bartlett SG: Spinach carbonic anhydrase primary structure deduced from the sequence of a cDNA clone. J Biol Chem 265: 5414–5417 (1990).

    Google Scholar 

  10. Felsentein J: Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783–791 (1985).

    Google Scholar 

  11. Fett JP, Coleman JR:; Characterization and expression of two cDNAs encoding carbonic anhydrase in Arabidopsis thaliana. Plant Physiol 105: 707–713 (1994).

    Google Scholar 

  12. Fukuzawa H, Fujiwara S, Tachiki A, Miyachi S: Nucleotide sequences of two genes CAH1 and CAH2 which encode carbonic anhydrase polypeptides in Chlamydomonas reinhardtii. Nucl Acids Res 18: 6441–6442 (1990).

    Google Scholar 

  13. Fukuzawa H, Suzuki E, Komukai Y, Miyachi S: A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon dioxide fixation by Synechococcus PCC7942. Proc Natl Acad Sci USA 89: 4437–4441 (1992).

    Google Scholar 

  14. Gutierrez M, Huber SC, Ku SB, Kanai R, Edwards GE: Intracellular location of carbon metabolism in the mesophyll cells of C4 plants. In: Avron M (ed) Proceedings of the Third International Congress on Photosynthesis, pp. 1219–1230. Elsevier, Amsterdam (1974).

    Google Scholar 

  15. Harlow E, Lane D: Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  16. Hatch MD, Burnell JN: Carbonic anhydrase activity in leaves and its role in the first step of C4 photosynthesis. Plant Physiol 93: 825–828 (1990).

    Google Scholar 

  17. Jacobson BS, Fong F, Heath RL: Carbonic anhydrase of spinach: Studies on its location, inhibition, and physiological function. Plant Physiol 55: 468–474 (1975).

    Google Scholar 

  18. Ku SB, Edwards GE: Photosynthesis in mesophyll protoplasts and bundle sheath cells of various types of C4 plants. Z Pflanzenphysiol 77: 16–32 (1975).

    Google Scholar 

  19. Ku MSB, Wu J, Dai Z, Scott RA, Chu C, Edwards GE: Photosynthetic and photorespiratory characteristics of Flaveria species. Plant Physiol 96: 518–528 (1991).

    Google Scholar 

  20. Majeau N, Coleman JR: Isolation and characterization of a cDNA coding for pea chloroplastic carbonic anhydrase. Plant Physiol 95: 264–268 (1991).

    Google Scholar 

  21. Majeau N, Coleman JR: Nucleotide sequence of a complementary DNA encoding tobacco chloroplastic carbonic anhydrase. Plant Physiol 100: 1077–1078 (1992).

    Google Scholar 

  22. Moore PD: Evolution of photosynthetic pathways in flowering plants. Nature 295: 647–648 (1982).

    Google Scholar 

  23. Okabe K, Yang S-Y, Tsuzuki M, Miyachi S: Carbonic anhydrase: Its content in spinach leaves and its taxonomic diversity studied with anti-spinach leaf carbonic anhydrase antibody. Plant Sci Lett 33: 145–153 (1984).

    Google Scholar 

  24. Olmsted JB: Analysis of cytoskeletal structures using blot-purified monospecific antibodies. Meth Enzymol 134: 467–472 (1986).

    Google Scholar 

  25. Poincelot RP: Intracellular distribution of carbonic anhydrase in spinach leaves. Biochim Biophys Acta 258: 637–642 (1972).

    Google Scholar 

  26. Provart NJ, Majeau N, Coleman JR: Characterization of pea chloroplastic carbonic anhydrase. Expression in Escherichia coli and site-directed mutagenesis. Plant Mol Biol 22: 937–943 (1993).

    Google Scholar 

  27. Raines CA, Horsnell PR, Holder C, Lloyd JC: Arabidopsis thaliana carbonic anhydrase: cDNA sequence and effect of CO2 on mRNA levels. Plant Mol Biol 20: 1143–1148 (1992).

    Google Scholar 

  28. Reed ML: Intracellular location of carbonate dehydratase (carbonic anhydrase) in leaf tissue. Plant Physiol 63: 216–217 (1979).

    Google Scholar 

  29. Reed ML, Graham D: Carbonic anhydrase in plants: Distribution, properties and possible physiological roles. In: Reinhold L, Harborne JB, Swain T (eds) Progress in Phytochemistry, vol 7, pp. 47–94. Pergamon Press, Oxford (1981).

    Google Scholar 

  30. Roeske CA, Ogren WL: Nucleotide sequence of pea cDNA encoding chloroplast carbonic anhydrase. Nucl Acids Res 18: 3413 (1990).

    Google Scholar 

  31. Saitou N, Nei M: The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406–425 (1987).

    Google Scholar 

  32. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  33. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  34. Sheen J-Y, Bogorad L: Differential expression of the ribulose bisphosphate carboxylase large subunit geneisin bundle sheath and mesophyll cells of developing maize leaves is influenced by light. Plant Physiol 79: 1072–1076 (1985).

    Google Scholar 

  35. Sung Y, Fuchs JA: Characterization of the cyn operon in Escherichia coli K12. J Biol Chem 263: 14769–14775 (1988).

    Google Scholar 

  36. Suzuki S, Burnell JN: Nucleotide sequence of a cDNA encoding rice chloroplastic carbonic anhydrase. Plant Physiol 107: 299–300 (1995).

    Google Scholar 

  37. Thompson JD, Higgins DG, Gibson TJ: CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucl Acids Res 22: 4673–4680 (1994).

    Google Scholar 

  38. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354 (1979).

    Google Scholar 

  39. Tsuzuki M, Miyachi S, Edwards GE: Localization of carbonic anhydrase in mesophyll cells of terrestrial C3 plants in relation to CO2 assimilation. Plant Cell Physiol 26: 881–891 (1985).

    Google Scholar 

  40. Utsunomiya E, Muto S: Carbonic anhydrase in the plasma membranes from leaves of C3 and C4 plants. Physiol Plant 88: 413–419 (1993).

    Google Scholar 

  41. von Heijne G, Steppuhn J, Herrmann RG: Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem 180: 535–545 (1989).

    Google Scholar 

Download references

Author information

Author notes

  1. Martha Ludwig

    Present address: Research School of Biological Sciences, Australian National University, P.O. Box 475, 2601, Canberra, ACT, Australia

  2. James N. Burnell

    Present address: Department of Molecular Sciences, James Cook University of Northern Queensland, 4814, Townsville, QLD, Australia

Authors and Affiliations

  1. Centre for Molecular Biotechnology, Queensland University of Technology, Box 2434, 4001, Brisbane, QLD, Australia

    Martha Ludwig & James N. Burnell

Authors
  1. Martha Ludwig
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. James N. Burnell
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ludwig, M., Burnell, J.N. Molecular comparison of carbonic anhydrase from Flaveria species demonstrating different photosynthetic pathways. Plant Mol Biol 29, 353–365 (1995). https://doi.org/10.1007/BF00043658

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00043658

Key words

Search

Navigation