Skip to main content
Log in

Immunoelectron-microscopy localization of abscisic acid with colloidal gold on Lowicryl-embedded tissues of Chnopodium polyspermum L.

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Further study on the localization of abscisic acid (ABA) has been undertaken at the ultrastructural level in Chenopodium polyspermum L. Axillary-bud-bearing nodes on the main axis were fixed with soluble 1-(3-dimethylaminopropyl)-3 ethyl carbodiimide, then postfixed with paraformaldehyde and embedded in Lowicryl K4M at-20° C. Ultrathin sections mounted on grids were successively incubated with rabbit anti-ABA antibodies and with gold-labelled goat anti-rabbit anti-bodies (40 nm particle size). Control sections treated with preimmune rabbit serum and ABA-preabsorbed antibodies were devoid of label. The background staining was very low with this technique. Quantitative analysis of the immunolabelling showed that two main sites of ABA accumulation could be defined: first, plastids in cortical cells and vascular parenchyma cells associated with sieve elements and xylem vessels; second, the cell cytoplasm and nucleus in the axillary bud tip and in procambial strands. In vascular bundles, the cambial cells showed no immunoreactivity. These observations support the hypothesis for the cytoplasmic synthesis of ABA which is subsequently trapped in plastids as cells mature.

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

ABA:

abscisic acid

EDC:

1-(3-dimethyl-aminopropyl)-3-ethyl carbodiimide

GAR 40:

goat anti-rabbit antibodies labelled with colloidal gold of particle size 40 nm

IgG:

immunoglobulin G

References

  • Bendayan, M. (1984) Protein A-gold electron microscopic immunocytochemistry: methods, applications and limitations. J. Electron Microsc. Technol. 1, 243–270

    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. (Oxford) 126, 123–143

    Google Scholar 

  • Cowan, I.R., Raven, J.A., Hartung, W., Farquhar, G.D. (1982) A possible role for abscisic acid in coupling stomatal conductance and photosynthetic carbon metabolism in leaves. Aust. J. Plant Physiol. 9, 489–498

    Google Scholar 

  • Craig, S., Goodchild, D.J. (1982) Post-embedding immunolabelling. Some effects of tissue preparation on the antigenicity of plant proteins. Eur. J. Cell Biol. 28, 251–256

    Google Scholar 

  • Craig, S., Miller, C. (1984) LR white resin and improved ongrid immunogold detection of Vicilin, a pea seed storage protein. Cell Biol. Int. Rep. 8, 879–886

    Google Scholar 

  • De Mey, J. (1983) Colloïdal gold probes in immunocytochemistry. In: Immunocytochemistry: practical applications in pathology and biology, vol 1, pp. 82–113, Polak, J.M., Van Noorden, S., eds. Wright, Bristol

    Google Scholar 

  • Goodchild, D.J., Andersson, B., Anderson, J.M. (1985a) Immunocytochemical localization of polypeptides associated with the oxygen evolving system of photosynthesis. Eur. J. Cell. Biol. 36, 294–298

    Google Scholar 

  • Goodchild, D.J., Anderson, J.M., Andersson, B. (1985b) Immunocytochemical localization of the cytochrome b/f complex of chloroplast thylakoid membranes Cell Biol. Int. Rep. 9, 715–721

    Google Scholar 

  • Greenwood, J.S., Chrispeels, M.J. (1985) Immunocytochemical localization of phaseolin and phytohemagglutinin in the endoplasmic reticulum and Golgi complex of developing bean cotyledons. Planta 164, 295–302

    Google Scholar 

  • Harris, N., Croy, R.R.D. (1985) The major albumin protein from pea (Pisum sativum L.). Localization by immunocytochemistry. Planta 165, 522–526

    Google Scholar 

  • Hartung, W., Heilmann, B., Gimmler, H. (1981) Do chloroplasts play a role in abscisic acid synthesis? Plant Sci. Lett. 22, 235–242

    Google Scholar 

  • Herman, E.M., Shannon L.M. (1984) Immunocytochemical evidence for the involvement of Golgi apparatus in the deposition of seed lectin of Bauhinia purpura (Leguminosae). Protoplasma 121, 163–170

    Google Scholar 

  • Leroux, B., Maldiney, R., Miginiac, E., Sossountzov, L., Sotta, B. (1985) Comparative quantitation of abscisic acid in plant extracts by gas-liquid chromotography and an enzyme-linked immunosorbent assay using the avidin-biotin system. Planta 166, 524–529

    Google Scholar 

  • Maldiney, R., Leroux, B., Sabbagh, I., Sotta, B., Sossountzov, L., Miginiac, E. (1986) Enzyme immunoassay to quantify three phytohormones: auxin, abscisic acid and zeatin-riboside, using a biotin/avidin system. J. Immunol. Meth., in press

  • Milborrow, B.V. (1979) Antitraspirants and regulation of abscisic acid content. Aust. J. Plant Physiol. 6, 249–254

    Google Scholar 

  • Newman, G.R., Jasani, B., Williams, E.D. (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope. Histochem. J. 15, 543–555

    Google Scholar 

  • Rodriguez, D., Nicolas, G., Aldasoro, J.J., Hernandez-Nistal, J., Babiano, M.J., Matilla, A. (1985) Altered development of polysomal RNA activity in chick-pea (Cicer arietimum I.) embryonic axes. Effects of abscisic acid and temperature. Planta 164, 517–523

    Google Scholar 

  • Roth, J., Bendayan, M., Orci, L. (1978) Ultrastructural localization of intracellular antigens by the use of protein-A gold complex. J. Histochem. Cytochem. 26, 1074–1081

    Google Scholar 

  • Roth, J., Bendayan, M., Carlemalm, E., Villiger, W., Garavito, M. (1981) Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J. Histochem. Cytochem. 29, 663–671

    Google Scholar 

  • Shaw, P.J., Henwood, J.A. (1985) Immuno-gold localization of cytochrome f, light-harvesting complex, ATP synthase and ribulose 1,5-bisophosphate carboxylase/oxygenase. Planta 165, 333–339

    Google Scholar 

  • Sotta, B., Sossountzov, L., Maldiney, R., Sabbagh, I., Tachon, P., Miginiac, E. (1985) Abscisic acid localization by light microscopic immunohistochemistry in Chenopodium polyspermum L. J. Histochem. Cytochem. 33, 201–208

    Google Scholar 

  • Tomenius, K., Clapham, D., Oxelfelt P. (1983) Localization by immunogold cytochemistry of viral antigens in sections of plant cells infected with red clover mottle virus. J. Gen. Virol. 645, 2669–2678

    Google Scholar 

  • Walton, D.C. (1980) Biochemistry and physiology of abscisic acid. Annu. Rev. Plant. Physiol. 31, 453–489

    Google Scholar 

  • Yamamoto, N., Yasuda, K. (1977) Use of a water soluble carbodiimide as a fixing agent. Acta Histochem. Cytochem. 10, 14–37

    Google Scholar 

  • Zavala, M.E., Brandon, B.L. (1983) Localization of a phytohormone using immunocytochemistry. J. Cell Biol. 97, 1235–1239

    Google Scholar 

  • Zeevaart, J.A.D. (1977) Sites of abscisic acid synthesis and metabolism in Ricinus communis. L. Plant Physiol. 59, 788–791

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sossountzov, L., Sotta, B., Maldiney, R. et al. Immunoelectron-microscopy localization of abscisic acid with colloidal gold on Lowicryl-embedded tissues of Chnopodium polyspermum L.. Planta 168, 471–481 (1986). https://doi.org/10.1007/BF00392266

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00392266

Key words

Navigation