Skip to main content
Log in

Immunocytochemical localization of phosphoenolpyruvate carboxylase and photosynthetic gas-exchange characteristics in ears of Triticum durum Desf.

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The presence and distribution of phosphoenolpyruvate carboxylase (PEPCase) in the glumes and immature grains of durum wheat (Triticum durum Desf.) were studied by electron-microscopical immunolabeling of PEPCase with polyclonal antibodies followed by protein A-gold. Plants were grown under mediterranean field conditions and samples were obtained two weeks after anthesis. In the kernels, high gold label was associated with the unstained areas of the protein bodies of aleurone cells, whereas labeling in the cytoplasm and chloroplasts of the pericarp was slight, although significantly above the background. In the glumes, high gold label was only located in cytoplasmic granules (vesicles) of the mesophyll cells, although labeling in the cytoplasm and chloroplasts was also significantly above the background. These observations in immature kernels and glumes are in accordance with the anaplerotic role of this enzyme, as evidenced in C3 plants. Measurements of apparent photosynthesis and its O2 dependence and CO2 compensation concentration were made on ears and flag leaves of durum wheat. In addition, an analog of phosphoenolpyruvate, 3,3-dichloro-2-dihydroxy-phosphinoylmethyl-2-propenoate, was used to inhibit PEPCase and, thereby, to assess the contribution of the PEPCase to photosynthesis in detached ears. There was no effect of the inhibitor on the apparent photosynthesis of ears. Whereas inhibition of apparent photosynthesis by 210 mL · L−1 O2 in flag leaves was typical of C3 species, inhibition in ears was even greater. The CO2 compensation concentrations in different ear parts were similar to or higher than in flag leaves and the O2 dependence was also comparable (about 70%). Therefore, gas-exchange data give further support to the assumption that a C4 cycle is absent or limited to very low rates in ears of durum wheat.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

AP:

apparent photosynthesis

BSA:

bovine serum albumin

DCDP:

3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate

PBS:

phosphate-buffered saline

PEP:

phosphoenolpyruvate

PEPCase:

PEP carboxylase

Γ:

CO2 compensation concentration

References

  • Adams, C.A., Leung, F., Sun, S.S.M. (1986) Molecular properties of phosphoenolpyruvate carboxylase from C3, C3–C4 intermediate, and C4 Flaveria species. Planta 167, 218–225

    Google Scholar 

  • Aoyagi, K., Bassham, J.A. (1984) Pyruvate orthophosphate dikinase of C3 seeds and leaves as compared to the enzyme from maize. Plant Physiol. 75, 387–392

    Google Scholar 

  • Aoyagi, K., Bassham, J.A., Greene, F.C. (1984) Pyruvate orthophosphate dikinase gene expression in developing wheat seeds. Plant Physiol. 75, 393–396

    Google Scholar 

  • Araus, J.L., Santiveri, P., Bosch-Serra, D., Royo, C., Romagosa, I. (1992) Carbon isotope ratios in ear parts of Triticale. Influence of grain filling. Plant Physiol. 100, 1033–1035

    Google Scholar 

  • Araus, J.L., Brown, R.H., Febrero, A., Bort, J., Serret, M.D. (1993) Ear photosynthesis, carbon isotope discrimination and the contribution of respiratory CO2 to differences in grain mass in durum wheat. Plant Cell Environ. 16, 383–392

    Google Scholar 

  • Black, C.C. (1973) Photosynthetic carbon fixation in relation to net CO2 uptake. Annu. Rev. Plant Physiol. 24, 253–286

    Google Scholar 

  • Blanke, M.M., Lenz, F. (1989) Fruit photosynthesis. Plant Cell Environ. 12, 31–46

    Google Scholar 

  • Brown, R.H., Bouton, J.H., Evans, P.T., Maltes, H.E., Rigsby, L.L. (1985) Photosynthesis, morphology, leaf anatomy, and cytogenetics of hybrids between C3 and C3/C4 Panicum species. Plant Physiol. 77, 653–658

    Google Scholar 

  • Brown, R.H., Bassett, C., Cameron, R., Evans, P.T., Bouton, J.H., Black, C.C., Sternberg, L'O.R., DeNiro, M.J. (1986) Photosynthesis in F1 hybrids between C4 and C3–C4 species in Flaveria. Plant Physiol. 82, 211–217

    Google Scholar 

  • Brown, R.H., Byrd, G.T., Black, C.C. (1991) Assessing the degree of C4 photosynthesis in C3–C4 species using an inhibitor of phosphoenolpyruvate carboxylase. Plant Physiol. 97, 985–989

    Google Scholar 

  • Brown, R.H., Byrd, G.T., Black, C.C. (1992) Degree of C4 photosynthesis in C4 and C3–C4 Flaveria species and their hybrids II. Inhibition of apparent photosynthesis by a phosphoenolpyruvate carboxylase inhibitor. Plant Physiol. 100, 947–950

    Google Scholar 

  • Caley, C.Y., Duffus, C.M., Jeffcoat, B. (1990) Photosynthesis in the pericarp of developing wheat grains. J. Exp. Bot. 41, 303–307

    Google Scholar 

  • Cameron, R.G., Bassett, C.L. (1988) Inheritance of C4 enzymes associated with carbon fixation in Flaveria species. Plant Physiol. 88, 532–536

    Google Scholar 

  • Cameron, R.G., Bassett, C.L., Bouton, J.H., Brown, R.H. (1989) Transfer of C4 photosynthetic characters through hybridization of Flaveria species. Plant Physiol. 90, 1538–1545

    Google Scholar 

  • Carlemalm, E., Garavito, R.M., Villiger, W. (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J. Microsc. 126, 123–143

    Google Scholar 

  • Duffus, C.M., Rosie, R. (1973) Some enzyme activities associated with the chlorophyll containing layers of the immature barley pericarp. Planta 114, 219–226

    Google Scholar 

  • Fincher, G.B. (1989) Molecular and cellular biology associated with endosperm mobilization in germinating cereal grains. Annu. Rev. Plant. Physiol. Plant Mol. Biol. 40, 305–346

    Google Scholar 

  • Hubick, K., Farquhar, G.D. (1989) Carbon isotope discrimination and the ratio of carbon gained to water lost in barley cultivars. Plant Cell Environ. 12, 795–804

    Google Scholar 

  • Jenkins, C.L.D. (1989) Effects of the phosphoenolpyruvate carboxylase inhibitor 3,3-dichloro-2-(dihydroxy-phosphinoylmethyl) propenoate on photosynthesis: C4 selectivity and studies on C4 photosynthesis. Plant Physiol. 89, 1231–1237

    Google Scholar 

  • Jenkins, C.L.D., Harris, R.L.N., McFadden, H.G. (1987) 3,3-Dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate, a new, specific inhibitor of phosphoenolpyruvate carboxylase. Biochem. Int. 14, 219–226

    Google Scholar 

  • Kriedemann, P. (1966) The photosynthetic activity of the wheat ear. Ann. Bot. 30, 349–363

    Google Scholar 

  • Lacoste-Royal, G., Gibbs, S.P. (1987) Immunocytochemical localization of ribulose-1,5-bisphosphate carboxylase in the pyrenoid and thylakoid region of the chloroplast of Chlamydomonas reinhardtii. Plant Physiol. 83, 602–606

    Google Scholar 

  • Matsuoka, M., Hata, S. (1987) Comparative studies of phosphoenolpyruvate carboxylase from C3 and C4 plants. Plant Physiol. 85, 947–951

    Google Scholar 

  • Melzer, E., O'Leary, M.H. (1987) Anapleurotic CO2 fixation by phosphoenolpyruvate carboxylase in C3 plants. Plant Physiol. 84, 58–60

    Google Scholar 

  • Melzer, E., O'Leary, M.H. (1991) Aspartic-acid synthesis in C3 plants. Planta 185, 368–371

    Google Scholar 

  • Meyer, A.O., Kelly, G.J., Latzko, E. (1978) Pyruvate orthophosphate dikinase of immature wheat grains. Plant. Sci. Lett. 12, 35–40

    Google Scholar 

  • Meyer, A.O., Kelly, G.J., Latzko, E. (1982) Pyruvate orthophosphate dikinase from the immature grains of cereal grasses. Plant Physiol. 69, 7–10

    Google Scholar 

  • O'Leary, M.H. (1982) Phosphoenolpyruvate carboxylase: an enzymologist's view. Annu. Rev. Plant Physiol. 33, 297–315

    Google Scholar 

  • Perrot-Rechenmann, C., Vidal, J., Brulfert, J., Burlet, A., Gadal, P. (1982) A comparative immunocytochemical localization study of phosphoenolpyruvate carboxylase in leaves of higher plants. Planta 155, 24–30

    Google Scholar 

  • Reed, J.E., Chollet, R. (1985) Immunofluorescent localization of phosphoenolpyruvate carboxylase and ribulose 1,5-bisphosphate carboxylase/oxygenase proteins in leaves of C3, C4 and C3–C4 intermediate Flaveria species. Planta 165, 439–445

    Google Scholar 

  • Salisbury, F.B., Ross, C.W. (1991) Plant physiology. 4th edn. Wadsworth Publishing Company, Belmont California

    Google Scholar 

  • Simpson, R.J., Dalling, M.J. (1981) Nitrogen redistribution during grain growth in wheat (Triticum aestivum L.). Planta 151, 447–456

    Google Scholar 

  • Singal, H.R., Sheoran, I.S., Singh, R. (1986) In vitro enzyme activities and products of 14CO2 assimilation in flag leaf and ear parts of wheat (Triticum aestivum L.). Photosynthesis Res. 8, 113–122

    Google Scholar 

  • Singh, U. (1981) The influence of irrigation on the composition of soluble amino acids in developing wheat grain. J. Sci. Food Agric. 32, 203–207

    Google Scholar 

  • von Caemmerer, S. (1992) Carbon isotope discrimination in C3–C4 intermediates: commissioned review. Plant Cell Environ. 15, 1063–1072

    Google Scholar 

  • Watson, P.A., Duffus, C.M. (1988) Carbon dioxide fixation by detached cereal caryopses. Plant Physiol. 87, 504–509

    Google Scholar 

  • Willmer, C.M., Roksandic, Z. (1980) Carbon isotope discrimination by tissues and organs from oat and barley inflorescences. J. Exp. Bot. 31, 1493–1495

    Google Scholar 

  • Wirth, E., Kelley, G.J., Fischbeck, G., Latzko, E. (1977) Enzyme activities and products of CO2 fixation in various photosynthetic organs of wheat and oat. Z Pflanzenphysiol. 82, 78–87

    Google Scholar 

  • Ziegler-Jöns, A. (1989) Gas-exchange of ears of cereals in response to carbon dioxide and light. II. Occurrence of a C3–C4 intermediate type of photosynthesis. Planta 178, 164–175

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

We thank Dr. Sam Sun (University of Hawaii) for kindly providing the PEPCase antibodies and Drs. C.L.D. Jenkins and M.D. Hatch (Division of Plant Industry, CSIRO, Canberra, Australia) for supplying DCDP for these experiments. We are also grateful to Dr. Robert Bargalló and Mrs. Ana Ribero (Servei de Microscopia Electronica, Universitat de Barcelona) for technical assistance in electron-microscopy studies. Participation of Drs. J.L. Araus and J. Bort in this work was supported by the Research Project of CICYT AGF92-0301, Spain.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Araus, J.L., Bort, J., Brown, R.H. et al. Immunocytochemical localization of phosphoenolpyruvate carboxylase and photosynthetic gas-exchange characteristics in ears of Triticum durum Desf.. Planta 191, 507–514 (1993). https://doi.org/10.1007/BF00195752

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195752

Key words

Navigation