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.
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
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
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
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
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
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
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
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
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
Harris, N., Croy, R.R.D. (1985) The major albumin protein from pea (Pisum sativum L.). Localization by immunocytochemistry. Planta 165, 522–526
Hartung, W., Heilmann, B., Gimmler, H. (1981) Do chloroplasts play a role in abscisic acid synthesis? Plant Sci. Lett. 22, 235–242
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
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
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
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
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
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
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
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
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
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
Walton, D.C. (1980) Biochemistry and physiology of abscisic acid. Annu. Rev. Plant. Physiol. 31, 453–489
Yamamoto, N., Yasuda, K. (1977) Use of a water soluble carbodiimide as a fixing agent. Acta Histochem. Cytochem. 10, 14–37
Zavala, M.E., Brandon, B.L. (1983) Localization of a phytohormone using immunocytochemistry. J. Cell Biol. 97, 1235–1239
Zeevaart, J.A.D. (1977) Sites of abscisic acid synthesis and metabolism in Ricinus communis. L. Plant Physiol. 59, 788–791
Author information
Authors and Affiliations
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392266