Photosynthetica, 1995 (vol. 33), issue 1

Photosynthetica 1995, 33(1):39-50 | DOI: 10.1023/A:1022190720904

In situ immunofluorescent localization of ribulose-1,5-bisphosphate carboxylase/oxygenase in mesophyll of C4 dicotyledonous plants

M. Castrillo1, P. Aso2, M. Longart3, A. Vermehren3
1 Depto Biología de Organismos, Universidad Simón Bolívar, Apdo, Caracas, Venezuela
2 Depto Biología Celular, Universidad Simón Bolívar, Apdo, Caracas, Venezuela
3 Coordinación de Biología, Universidad Simón Bolívar, Apdo, Caracas, Venezuela

The location of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) in the leaf mesophyll of some dicotyledonous C4 plants was confirmed by immunofluorescent labelling. The anti-RuBPCO immune serum was obtained by inoculating a rabbit with commercially obtained RuBPCO. Specificity of these antibodies was tested by immunodiffusion, immunoelectrophoresis, and Western blotting. Fresh hand-cuts of leaves from dicotyledonous C4 plants, Amaranthus caudatus, A. dubius, Gomphrena globosa, and Portulaca oleracea, were incubated with the conjugated anti-RuBPCO immune serum and then with a commercial FITC-anti-rabbit IgG conjugate. Nerium oleander was used a control C3 plant pattern and Zea mays as a C4 plant pattern. The immunofluorescent label was distributed in both mesophyll and bundle sheath in all the C4 plants tested. It is an unequivocal proof that in the C4 dicotyledonous plants the RuBPCO is not only located in the chloroplasts of the bundle sheath cells but also in the chloroplasts of the mesophyll cells. In these plants therefore, the C4 pathway cannot exclusively be viewed as an intercellular level concentration mechanism. In the mesophyll cytoplasm, phosphoenolpyruvate carboxylase traps CO2, while in the mesophyll chloroplasts, RuBPCO operates with atmospheric CO2 and CO2 from the C4 decarboxylation step at an intracellular level, which could mean a significant energetic economy. The CO2 from photorespiration could be saved and reincorporated. Location of RuBPCO in the mesophyll and/or bundle sheath chloroplasts is a matter of inter- and intracellular compartmentation which makes another variation of C4 photosynthetic pathway possible.

Keywords: Amaranthus; Gomphrena; Portulaca; Nerium; Zea

Prepublished online: March 1, 1997; Published: March 1, 1998Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Castrillo, M., Aso, P., Longart, M., & Vermehren, A. (1995). In situ immunofluorescent localization of ribulose-1,5-bisphosphate carboxylase/oxygenase in mesophyll of C4 dicotyledonous plants. Photosynthetica33(1), 39-50. doi: 10.1023/A:1022190720904.
Download citation

References

  1. Bassüner, B., Keerberg, O., Bauwe, H., Pyarnik, T., Keerberg, H.: Photosynthetic CO2 metabolism in C3-C4 intermediate and C4 species of Flaveria (Asteraceae).-Biochem. Physiol. Pflanzen 179: 631-634, 1984. Go to original source...
  2. Bauwe, H.: Photosynthetic enzyme activities and immunofluorescence studies on the localization of ribulose-1,5-bisphosphate carboxylase/oxygenase in leaves of C3, C4 and C3-C4 intermediate species of Flaveria (Asteraceae).-Biochem. Physiol. Pflanzen 179: 235-268, 1984. Go to original source...
  3. Black, C.C., Jr.: Photosynthetic carbon fixation in relation to net CO2 uptake.-Annu. Rev. Plant Physiol. 24: 253-286, 1973. Go to original source...
  4. Brown, R.H.: Photosynthesis of grass species differing in carbon dioxide fixation pathways. IV. Analysis of reduced oxygen response in Panicum milioides and Panicum schenckii.-Plant Physiol. 65: 346-349, 1980. Go to original source...
  5. Brown, R.H., Hattersley, P.W.: Leaf anatomy of C3-C4 species as related to evolution of C4 photosynthesis.-Plant Physiol. 91: 1543-1550, 1990
  6. Castrillo, M., Whatley, F.R.: A differential mechanical technique for tissue separation in dicotyledonous C4 plants.-Photosynthetica 30: 269-277, 1994.
  7. Cheng, S.-H., Moore, B.d., Edwards, G.E., Ku, M.S.B.: Photosynthesis in Flaveria brownii, a C4-like species. Leaf anatomy, characteristics of CO2 exchange, compartmentation of photosynthetic enzymes, and metabolism of 14CO2.-Plant Physiol. 87: 867-873, 1988. Go to original source...
  8. Cheng, S.-H., Moore, B.d., Wu, J., Edwards, G.E., Ku, M.S.B.: Photosynthetic plasticity in Flaveria brownii. Growth irradiance and the expression of C4 photosynthesis.-Plant Physiol. 89: 1129-1135, 1989. Go to original source...
  9. Dengler, N.G., Dengler, R.E., Grenville, D.J.: Comparison of photosynthetic carbon reduction (Kranz) cells having different ontogenetic origins in the C4 NADP-malic enzyme grass Arundinella hirta.-Can. J. Bot. 68: 1222-1232, 1990. Go to original source...
  10. Dengler, R.E., Dengler, N.G.: Leaf vascular architecture in the atypical NADP-malic enzyme grass Arundinella hirta.-Can. J. Bot. 68: 1208-1221, 1990
  11. Edwards, G.E., Sheta, E., Moore, B.d., Dai, Z., Franceschi, V.R., Cheng, S.H., Lin, C.-H., Ku, M.S.B.: Photosynthetic characteristics of cassava (Manihot esculenta Crantz), a C3 species with chlorenchymatous bundle sheath cells.-Plant Cell Physiol. 31: 1199-1206, 1990.
  12. Hatch, M.D.: C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure.-Biochim. biophys. Acta 895: 81-106, 1987. Go to original source...
  13. Hatch, M.D., Slack, C.R., Bull, T.A.: Light-induced changes in the content of some enzymes of the C4-dicarboxylic acid pathway of photosynthesis and its effect on other characteristics of photosynthesis.-Phytochemistry 8: 697-706, 1969. Go to original source...
  14. Hattersley, P.W., Watson, L., Osmond, C.B.: In situ immunofluorescent labelling of ribulose-1,5-bisphosphate carboxylase in leaves of C3 and C4 plants.-Aust. J. Plant Physiol. 4: 523-539, 1977.
  15. Hegde, B.A., Patil, T.M.: Parthenium hysterophorus (L.), a C3 plant with "Kranz" syndrome.-Photosynthetica 15: 1-4, 1981.
  16. Holaday, A.S., Brown, R.H., Bartlett, J.M., Sandlin, E.A., Jackson, R.C.: Enzymic and photosynthetic characteristics of reciprocal F1 hybrids of Flaveria pringlei (C3) and Flaveria brownii (C4-like species).-Plant Physiol. 87: 484-490, 1988. Go to original source...
  17. Kennedy, R.A.: Photorespiration in C3 and C4 plant tissue cultures. Significance of Kranz anatomy to low photorespiration in C4 plants.-Plant Physiol. 58: 573-575, 1976. Go to original source...
  18. Matsumoto, K., Nishimura, M., Akazawa, T.: Ribulose-1,5-bisphosphate carboxylase in the bundle sheath cells of maize leaf.-Plant Cell Physiol. 18: 1281-1290, 1977. Go to original source...
  19. Monson, R.K., Edwards, G.E., Ku, M.S.B.: C3-C4 intermediate photosynthesis in plants.-BioScience 34: 563-566, 571-574, 1984. Go to original source...
  20. Moore, P.D.: Evolution of photosynthetic pathway in flowering plants.-Nature 295: 647-648, 1982. Go to original source...
  21. Nelson, T., Langdale, J.A.: Developmental genetics of C4 photosynthesis.-Annu. Rev. Plant Physiol. Plant mol. Biol. 43: 25-47, 1992. Go to original source...
  22. Ouchterlony, O.: Diffusion-in-gel methods for immunological analysis.-Progr. Allergy 5: 1-78, 1958. Go to original source...
  23. Perrot-Rechenmann, C., Jacquot, J.P., Gadal, P., Weeden, N.F., Cséke, C., Buchanan, B.B.: Localization of NADP-malate dehydrogenase of corn leaves by immunological methods.-Plant Sci. Lett. 30: 219-226, 1983. Go to original source...
  24. Perrot-Rechenmann, C., Vidal, J., Brulfert, J., Burlet, A., Gadal, P.: A comparative immunocytochemical localization study of phosphoenolpyruvate carboxylase in leaves of higher plants.-Planta 155: 24-30, 1982. Go to original source...
  25. Reed, J.E., Chollet, R.: 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, 1985. Go to original source...
  26. Reiskind, J.B., Berg, R.H., Salvucci, M.E., Bowes, G.: Immunogold localization of primary carboxylases in leaves of aquatic and a C3-C4 intermediate species.-Plant Sci. 61: 43-52, 1989. Go to original source...
  27. Reyes-Fernández, M.E., Baker, D.A.: Immunogold localization of ribulose-1,5-bisphosphate carboxylase/oxygenase and glycine decarboxylase in the "non-classical" C4 species Aristida funiculata.-Photosynthetica 31: 379-388, 1995.
  28. Sheen, J.Y., Bogorad, L.: Differential expression of the ribulose bisphosphate carboxylase large subunit gene in bundle sheath and mesophyll cells of developing maize leaves is influenced by light.-Plant Physiol. 79: 1072-1076, 1985. Go to original source...
  29. Shomer-Ilan, A., Beer, S., Waisel, Y.C.: Suaeda monoica, a C4 plant without typical bundle sheath.-Plant Physiol. 56: 676-679, 1985.