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The release of α-amylase through gibberellin-treated barley aleurone cell walls

An immunocytochemical study with Lowicryl K4M

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Abstract

Localisation of α-amylase (EC 3.2.1.1.) in low-temperature-embedded isolated barley (Hordeum vulgare L.) aleurone has been achieved using rhodamine-labelled secondary antibodies and the protein A-gold technique. Treatment with gibberellic acid (GA3) resulted in an increase of immunofluorescence in the cytoplasm of aleurone cells and also its appearance in specific regions of the cell walls. Cytoplasmic label was neither perinuclear nor associated specifically with aleurone grains as had been found in earlier work, but was present throughout the cytoplasm of all cells. A relatively high level of labelling occurred in hydrolysed wall regions. Label was also associated with plasmodesmata in both hydrolysed and unhydrolysed wall regions. The pattern of labelling indicates that α-amylase is released from aleurone via digested wall channels and that, except for the inner wall layer, unhydrolysed regions are impermeable to the enzyme. It is suggested that the resistant wall tubes around plasmodesmata may facilitate enzyme release by providing a pathway for transfer, especially of wall hydrolases, into the more impermeable parts of the wall.

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Abbreviations

ER:

endoplasmic reticulum

GA3 :

gibberellic acid

RER:

rough endoplasmic reticulum

References

  • Akazawa, T., Hara-Nishimura, I. (1985) Topographic aspects of biosynthesis, extracellular secretion, and intracellular storage of proteins in plant cells. Annu. Rev. Plant Physiol. 36, 441–472

    Google Scholar 

  • Ashford, A.E., Allaway, W.G., Gubler, F., Lennon, A., Sleegers, J. (1986) Temperature control in Lowicryl K4M and glycol methacrylate during polymerisation: is there a low-temperature embedding method? J. Microsc. 144, 107–126

    Google Scholar 

  • Ashford, A.E., Gubler, F. (1984) Mobilization of polysaccharide reserves from endosperm. In: Seed physiology, vol. 2: Germination and reserve mobilization, pp. 117–162, Murray, D.R., ed. Academic Press Sydney, Australia

    Google Scholar 

  • Ashford, A.E., Jacobsen, J.V. (1974) Cytochemical localization of phosphatase in barley aleurone cells: the pathway of gibberellic-acid-induced enzyme release. Planta 120, 81–105

    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 

  • Carpita, N.C. (1982) Limiting diameters of pores and the surface structure of plant cell walls. Science 218, 813–814

    Google Scholar 

  • Carpita, N., Sabularse, D., Montezinos, D., Delmer, D.P. (1979) Determination of the pore size of cell walls of living plant cells. Science 205, 1144–1147

    Google Scholar 

  • Chrispeels, M.J., Varner, J.E. (1967) Gibberellic acid-enhanced synthesis and release of α-amylase and ribonuclease by isolated barley aleurone layers. Plant Physiol. 42, 398–406

    Google Scholar 

  • Faye, L., Sturm, A., Bollini, R., Vitale, A., Chrispeels, M.J. (1986) The position of the oligosaccharide side-chains of phytohemagglutinin and their accessibility to glycosidases determines their subsequent processing in the Golgi. Eur. J. Biochem. 158, 655–661

    Google Scholar 

  • Filner, P., Varner, J.E. (1967) A test for de novo synthesis of enzymes: density labeling with H2O18 of barley α-amylase induced by gibberellic acid. Proc Natl. Acad. Sci. USA 58, 1520–1526

    Google Scholar 

  • Gardiner, M., Chrispeels, M.J. (1975) Involvement of the Golgi apparatus in the synthesis and secretion of hydroxyproline-rich cell wall glycoproteins. Plant Physiol. 55, 536–541

    Google Scholar 

  • Giloh, H., Sedat, J.W. (1982) Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science 217, 1252–1255

    Google Scholar 

  • Griffiths, G., Simons, K. (1987) The trans Golgi network: sorting at the exit site of the Golgi complex. Science 234, 438–443

    Google Scholar 

  • Goodwin, P.B. (1983) Molecular size limit for movement in the symplast of the Elodea leaf. Planta 157, 124–130

    Google Scholar 

  • Gubler, G., Ashford, A.E. (1985) Release of ferulic acid esters from barley aleurone. I. Time course of gibberellic-acid induced release from isolated layers. Aust. J. Plant. Physiol. 12, 297–305

    Google Scholar 

  • Gubler, F., Jacobsen, J.V., Ashford, A.E. (1986) Involvement of the Golgi apparatus in the secretion of α-amylase from gibberellin-treated barley aleurone cells. Planta 168, 447–452

    Google Scholar 

  • Gunning, B.E.S., Overall, R.L. (1983) Plasmodesmata and cell-to-cell transport in plants. BioScience 33, 260–265

    Google Scholar 

  • Heupke, H., Robinson, D.G. (1985) Intracellular transport of α-amylase in barley aleurone cells: evidence for the participation of the Golgi apparatus. Eur. J. Cell Biol. 39, 265–272

    Google Scholar 

  • Higgins, T.J.V., Jacobsen, J.V., Zwar, J.A. (1982) Gibberellic acid and abscisic acid modulate protein synthesis and mRNA levels in barley aleurone layers. Plant Mol. Biol. 1, 191–215

    Google Scholar 

  • Jacobsen, J.V., Knox, R.B. (1973) Cytochemical localization and antigenicity of α-amylase in barley aleurone tissue. Planta 112, 213–224

    Google Scholar 

  • Jacobsen, J.V., Zwar, J.A., Chandler, P.M. (1985) Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley. Planta 163, 430–438

    Google Scholar 

  • Jones, R.L. (1969a) Gibberellic acid and the fine structure of barley aleurone cells. I. Changes during the lag-phase of α-amylase synthesis. Planta 87, 119–133

    Google Scholar 

  • Jones, R.L. (1969b) Gibberellic acid and the fine structure of barley aleurone cells. II. Changes during the synthesis and secretion of α-amylase. Planta 88, 73–86

    Google Scholar 

  • Jones, R.L. (1972) Fractionation of the enzymes of the barley aleurone layer: evidence for a soluble mode of release. Planta 103, 95–109

    Google Scholar 

  • Jones, R.L. (1985) Protein synthesis and secretion by the barley aleurone: a perspective. Isr. J. Bot. 34, 377–395

    Google Scholar 

  • Jones, R.L., Chen, R. (1976) Immunohistochemical localization of α-amylase in barley aleurone cells. J. Cell. Sci. 20, 183–198

    Google Scholar 

  • Jones, R.L., Jacobsen, J.V. (1982) The role of the endoplasmic reticulum in the synthesis and transport of α-amylase in barley aleurone layers. Planta 156, 421–432

    Google Scholar 

  • Kawasaki, S., Sato, S. (1979) Isolation of the Golgi apparatus from suspension cultured tobacco cells and preliminary observations on the intracellular transport of extensin-precursor. Bot. Mag. 92, 305–314

    Google Scholar 

  • Kelly, R.B. (1985) Pathways of protein secretion in eukaryotes. Science 230, 25–32

    Google Scholar 

  • Osborn, M., Weber, K. (1982) Immunofluorescence and immunocytochemical procedures with affinity purified antibodies: tubulin-containing structures. In: Methods in cell biology, vol. 24: The cytoskeleton, pt. A, pp. 97–132, Wilson, I., ed. Academic Press, New York

    Google Scholar 

  • Overall, R.L., Wolfe, J., Gunning, B.E.S. (1982) Intercellular communication in Azolla roots: 1. Ultrastructure of plasmodesmata. Protoplasma 111, 134–150

    Google Scholar 

  • Pyliotis, N.A., Ashford, A.E., Whitecross, M.I., Jacobsen, J.V. (1979) Localization of gibberellic acid-induced acid phosphatase activity with the electron microscope. Planta 147, 134–140

    Google Scholar 

  • Rey, M.E.C., Noble, J.P. (1984) Subcellular localization by immunocytochemistry of the extracellular protease produced by Nectria galligena Bres. in infected apple tissue. Physiol. Plant Path. 25, 323–336

    Google Scholar 

  • Taiz, L., Jones, R.L. (1970) Gibberellic acid, β-1,3-glucanase and the cell walls of barley aleurone layers. Planta 92, 73–84

    Google Scholar 

  • Taiz, L., Jones, R.L. (1973) Plasmodesmata and an associated cell wall component in barley aleurone tissue. Am. J. Bot. 60, 67–75

    Google Scholar 

  • Valnes, K., Brandtzaeg, P. (1985) Retardation of immunofluorescence fading during microscopy. J. Histochem. Cytochem. 33, 755–761

    Google Scholar 

  • Van Noorden, S., Polak, J.M. (1983) Immunocytochemistry today techniques and practive. In: Immunocytochemistry: practical applications in pathology and biology, pp. 11–42, Polak, J.M., Van Noorden, S., eds. J. Wright, Bristol, UK

    Google Scholar 

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Authors and Affiliations

  1. School of Botany, University of New South Wales, P.O. Box 1, 2033, Kensington, N.S.W., Australia

    F. Gubler & A. E. Ashford

  2. CSIRO Division of Plant Industry, 2601, Canberra, A.C.T., Australia

    J. V. Jacobsen

Authors
  1. F. Gubler
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  2. A. E. Ashford
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  3. J. V. Jacobsen
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Gubler, F., Ashford, A.E. & Jacobsen, J.V. The release of α-amylase through gibberellin-treated barley aleurone cell walls. Planta 172, 155–161 (1987). https://doi.org/10.1007/BF00394583

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  • DOI: https://doi.org/10.1007/BF00394583

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