Skip to main content
Log in

Genomic structure and expression of the pyruvate, orthophosphate dikinase gene of the dicotyledonous C4 plant Flaveria trinervia (Asteraceae)

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

Abstract

Pyruvate orthophosphate dikinase (PPDK) is a key enzyme of C4 photosynthesis providing the acceptor molecule for the primary CO2 fixation in the mesophyll cells. Here we present the isolation and characterisation of the corresponding gene (termed pdk) from the C4 plant Flaveria trinervia (Asteraceae). Southern analysis indicates that in contrast to maize pdk sequences in F. trinervia are present as single copy. Sequence analysis of the entire gene reveals that its coding sequence is identical to the previous isolated PPDK-cDNA from this species. The gene spans about 13 kb and consists of 21 exons, it thus contains two additional exons compared to the maize gene. As in maize, a long intervening sequence of 6.1 kb is positioned at the boundary of the transit peptide segment and the mature protein region. Pdk transcripts accumulate abundantly in leaves, but are also detectable in stems and roots. While the leaf and stem transcripts are 3.4 kb in size and encode the chloroplastic PPDK isoform, a 3.0 kb transcript lacking the region encoding the plastidic transit peptide accumulates in roots. Thus two different transcripts can be produced from a single pdk gene most likely by use of alternative promoters and not by alternative splicing. The accumulation of the 3.4 kb transcript is under light control. Darkening leads to a drastic depletion of this transcript in both leaves and stems. Instead, the 3.0 kb transit peptide-lacking pdk transcript accumulates, but only in stems and roots, not in leaves.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Aoyagi K, Bassham JA: Pyruvate orthophosphate dikinase in wheat leaves. Plant Physiol 73: 853–854 (1983).

    Google Scholar 

  2. Aoyagi K, Bassham JA: Pyruvate orthophosphate dikinase mRNA organ specificity in wheat and maize. Plant Physiol 76: 278–280 (1984).

    Google Scholar 

  3. Aoyagi K, Kuhlemeier C, Chua NH: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).

    Google Scholar 

  4. Aoyagi K, Nakamoto H: Pyruvate,Pi dikinase in bundle sheath strands as well as in mesophyll cells in maize leaves. Plant Physiol 78: 661–664 (1985).

    Google Scholar 

  5. Aviv H, Leder P: Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69: 1408–1412 (1972).

    Google Scholar 

  6. Bruchhaus I, Tannich E: Primary structure of the pyruvate phosphate dikinase in Entamoeba histolytica. Mol Biochem Parasitol 62: 153–156 (1993).

    Google Scholar 

  7. Chen EY, Seeburg PH: Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA 4: 165–170 (1985).

    Google Scholar 

  8. Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).

    Google Scholar 

  9. Cockburn W: Stomatal mechanism as the basis of the evolution of CAM and C4 photosynthesis. Plant Cell Environ 6: 275–279 (1983).

    Google Scholar 

  10. Cole KP, Blakeley SD, Dennis DT: Structure of the gene encoding potato cytosolic pyruvate kinase. Gene 122: 255–261 (1992).

    Google Scholar 

  11. Cushman JC, Meyer G, Michalowski CB, Schmitt JM, Bohnert HJ: Salt stress leads to differential expression of two isogenes of phosphoenolpyruvate carboxylase during crassulancean acid metabolism induction in the common ice plant. Plant Cell 1: 715–725 (1989).

    Google Scholar 

  12. Darnell JE, Doolittle WF: Speculations on the early course of evolution. Proc Natl Acad Sci USA 83: 1271–1275 (1986).

    Google Scholar 

  13. Dorit RL, Gilbert W: The limited universe of exons. Curr Opin Genet Devel 1: 464–469 (1991).

    Google Scholar 

  14. Edwards GE, Ku MSB. Biochemistry of C3-C4 intermediates. In: Hatch MD, Boardman NK (eds) The Biochemistry of Plants, vol. 10, pp 275–325. Academic Press, Inc., New York (1987).

    Google Scholar 

  15. Edwards GE, Nakamoto H, Burnell JN, Hatch MD: Pyruvate,Pi dikinase and NADP-malate dehydrogenase in C4 photosynthesis: properties and mechanism of light/dark regulation. Annu Rev Plant Physiol 36: 255–286 (1985).

    Google Scholar 

  16. Edwards GE, Walker DA. C3, C4: Mechanism, and Cellular and Environmental Regulation, of Photosynthesis. Blackwell Scientific Publications, Oxford London (1983).

    Google Scholar 

  17. Ehleringer JR, Sage RF, Flanagan LB, Pearcy RW. Climate change and the evolution of C4 photosynthesis. Trends Ecol Evol 6: 95–99 (1991).

    Google Scholar 

  18. Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    Google Scholar 

  19. Glackin CA, Grula JW: Organ-specific transcripts of different size and abundance derive from the same pyruvate, orthophosphate dikinase gene in maize. Proc Natl Acad Sci USA 87: 3004–3008 (1990).

    Google Scholar 

  20. Hanley BA, Schuler MA: Plant intron sequences: evidence for distinct groups of introns. Nucl Acids Res 16: 7159–7174 (1988).

    Google Scholar 

  21. Harvey PH, Purvis A: Comparative methods for explaining adaptations. Nature 351: 619–624 (1991).

    Google Scholar 

  22. Hata S, Matsuoka M: Immunological studies on pyruvate orthophosphate dikinase in C3 plants. Plant Cell Physiol 28: 635–641 (1987).

    Google Scholar 

  23. Hatch MD: C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim Biophys Acta 895: 81–106 (1987).

    Google Scholar 

  24. Hattersley PW. Variations in photosynthetic pathway. In: Soderstrom TR, Hilu KW, Campbell CS, Barkworth ME (eds) Grass Systematics and Evolution, pp. 49–64. Smithonian Institution Press, Washington (1987).

    Google Scholar 

  25. Helentjaris T, Weber D, Wright S: Identification of the genomic locations of duplicate nucleotide sequences in maize by analysis of restriction fragment length polymorphisms. Genetics 118: 353–363 (1988).

    Google Scholar 

  26. Henikoff S: Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).

    Google Scholar 

  27. Hermans J, Westhoff P: Analysis of expression and evolutionary relationships of phosphoenolpyruvate carboxylase genes in Flaveria trinervia (C4) and F. pringlei (C3). Mol Gen Genet 224: 459–468 (1990).

    Google Scholar 

  28. Hermans J, Westhoff P: Homologous genes for the C4 isoform of phospheenolpyruvate carboxylase in a C3-and a C4-Flaveria species. Mol Gen Genet 234: 275–284 (1992).

    Google Scholar 

  29. Hudspeth RL, Glackin CA, Bonner J, Grula JW: Genomic and cDNA clones for maize phosphoenolpyruvate carboxylase and pyruvate,orthophosphate dikinase: expression of different gene-family members in leaves and roots. Proc Natl Acad Sci USA 83: 2884–2888 (1986).

    Google Scholar 

  30. Höfer MU, Santore UJ, Westhoff P: Differential accumulation of the 10-, 16- and 23-kDa peripheral components of the water-splitting complex of photosystem II in mesophyll and bundle-sheath chloroplasts of the dicotyledonous C4 plant Flaveria trinervia (Spreng.) C. Mohr. Planta 186: 304–312 (1992).

    Google Scholar 

  31. Lepiniec L, Keryer E, Philippe H, Gadal P, Crétin C: Sorghum phosphoenolpyruvate carboxylase gene family: Structure, function and molecular evolution. Plant Mol Biol 21: 487–502 (1993).

    Google Scholar 

  32. Lipka B, Steinmüller K, Rosche E, Börsch D, Westhoff P. The C3 plant Flaveria pringlei contains a plastidic NADP-malic enzyme which is orthologous to the C4 iso-form of the C4 plant F. trinervia. Plant Mol Biol 26: 1775–1783 (1994).

    Google Scholar 

  33. Lou H, McCullough AJ, Schuler MA: 3′ Splice site selection in dicot plant nuclei is position dependent. Mol Cell Biol 13: 4485–4493 (1993).

    Google Scholar 

  34. Matsuoka M: Structure, genetic mapping, and expression of the gene for pyruvate, orthophosphate dikinase from maize. J Biol Chem 265: 16772–16777 (1990).

    Google Scholar 

  35. Matsuoka M, Ozeki Y, Yamamoto N, Hirano H, Kano-Murakami Y, Tanaka Y: Primary structure of maize pyruvate, orthophosphate dikinase as deduced from cDNA sequence. J Biol Chem 263: 11080–11083 (1988).

    Google Scholar 

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

    Google Scholar 

  37. Norris SR, Meyer SE, Callis J: The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression. Plant Mol Biol 21: 895–906 (1993).

    Google Scholar 

  38. Ohta T: Multigene families and the evolution of complexity. J Mol Evol 33: 34–41 (1991).

    Google Scholar 

  39. Oswald A, Streubel M, Ljungberg U, Hermans J, Eskins K, Westhoff P: Differential biogenesis of photosystem II in mesophyll and bundle sheath cells of NADP malic enzyme-type C4 plants. A comparative protein and RNA analysis. Eur J Biochem 190: 185–194 (1990).

    Google Scholar 

  40. Palmer JD, Logsdon JM: The recent origin of introns. Curr Opin Genet Devel 1: 470–477 (1991).

    Google Scholar 

  41. Pokalyco DJ, Carroll LJ, Martin MB, Babbitt PC, Cunaway-Mariano D: Analysis of sequence homologies in plant and bacterial Pyruvate Phosphate Dikinase, enzyme I of the bacterial phosphoenolpyruvate: sugar phosphotransferase system and other PEP-utilizing enzymes. Identification of potential catalytic and regulatory motifs. Biochemistry 29: 10757–10765 (1990).

    Google Scholar 

  42. Powell AM: Systematics of Flaveria (Flaveriinae-Asteraceae). Ann Mo Bot Gard 65: 590–636 (1978).

    Google Scholar 

  43. Raghavendra AS: Characteristics of plant species intermediate between C3 and C4 pathways of photosynthesis: their focus of mechanism and evolution of C4 syndrome. Photosynthetica 14: 271–283 (1980).

    Google Scholar 

  44. Rosche E, Streubel M, Westhoff P: Primary structure of the photosynthetic pyruvate orthophosphate dikinase of the C3 plant Flaveria pringlei and expression analysis of pyruvate orthophosphate dikinase sequences in C3, C3-C4 and C4 Flaveria species. Plant Mol Biol 26: 763–769 (1994).

    Google Scholar 

  45. Rosche E, Westhoff P: Primary structure of pyruvate, orthophosphate dikinase in the dicotyledonous C4 plant Flaveria trinervia. FEBS Lett 273: 116–121 (1990).

    Google Scholar 

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

    Google Scholar 

  47. Schäffner AR, Sheen J: Maize C4 photosynthesis involves differential regulation of phosphoenolpyruvate carboxylase genes. Plant J 2: 221–232 (1992).

    Google Scholar 

  48. Sheen J: Molecular mechanisms underlying the differential expression of maize pyruvate, orthophosphate dikinase genes. Plant Cell 3: 225–245 (1991).

    Google Scholar 

  49. Steinmüller K, Batschauer A, Apel K: Tissue-specific and light-dependent changes of chromatin organization in barley (Hordeum vulgare). Eur J Biochem 158: 519–525 (1986).

    Google Scholar 

  50. Stockhaus J, Poetsch W, Steinmüller K, Westhoff P: Evolution of the C4 phosphoenolpyruvate carboxylase promoter of the C4 dicot Flaveria trinervia: an expression analysis in the C3 plant tobacco. Mol Gen Genet 245: 286–293 (1994).

    Google Scholar 

  51. Tabor S, Richardson CC: Effect of manganese ions on the incorporation of dideoxynucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase. I. Proc Natl Acad Sci USA 86: 4076–4080 (1989).

    Google Scholar 

  52. Thompson WF, White MJ: Physiological and molecular studies of light-regulated nuclear genes in higher plants. Annu Rev Plant Physiol Plant Mol Biol 42: 423–466 (1991).

    Google Scholar 

  53. Tobin EM, Silverthorne J: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).

    Google Scholar 

  54. Westhoff P, Herrmann RG: Complex RNA maturation in chloroplasts. The psbB operon from spinach. Eur J Biochem 171: 551–564 (1988).

    Google Scholar 

  55. Westhoff P, Offermann-Steinhard K, Höfer M, Eskins K, Oswald A, Streubel M: Differential accumulation of plastid transcripts encoding photosystem II components in the mesophyll and bundle-sheath cells of monocotyledonous NADP-malic enzyme-type C4 plants. Planta 184: 377–388 (1991).

    Google Scholar 

  56. Whitkus R, Doebley J, Lee M: Comparative genome mapping of sorghum and maize. Genetics 132: 1119–1130 (1992).

    Google Scholar 

  57. Wilson AC, Carlson SS, White TJ: Biochemical evolution. Annu Rev Biochem 46: 573–639 (1977).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rosche, E., Westhoff, P. Genomic structure and expression of the pyruvate, orthophosphate dikinase gene of the dicotyledonous C4 plant Flaveria trinervia (Asteraceae). Plant Mol Biol 29, 663–678 (1995). https://doi.org/10.1007/BF00041157

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00041157

Key words

Navigation