Skip to main content
Log in

Microtubules and coated vesicles in guard-cell protoplasts ofAllium cepa L.

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Protoplasts were prepared from the guard cells ofA. cepa. Epidermal peels taken from expanding green leaves and largely free of mesophyll were treated with Cellulysin, and protoplasts were harvested after 18 h of digestion. That the protoplasts were derived from guard cells was ascertained from their characteristic vacuolar autofluorescence and from observations showing that all other epidermal cells are killed in the peeling procedure. The protoplasts proved to be a good system with which to view the cell cortex and inner surface of the plasmalemma. The lysis of cells adhering to polylysine-treated, Formvar-coated grids, followed by negative staining in uranyl acetate, showed that many microtubules normally present in ordered arrays in situ remain closely applied to the inner surface of the plasmalemma in protoplasts. In addition, numerous vesiculate elements including coated vesicles and/or pits are present amongst the microtubules. Similar vesicles are evident in thin sections of fixed, embedded guard cells and protoplasts. The significance of these structures in the cell cortex is discussed.

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

References

  • Albertini, D.F., Clark, J.I. (1975) Membrane-microtubule interactions: concanavalin A capping induced redistribution of cytoplasmic microtubules and colchicine binding proteins. Proc. Natl. Acad. Sci. USA72, 4976–4980

    Google Scholar 

  • Allen, N.S., Allen, R.D. (1978) Cytoplasmic streaming in green plants. Annu. Rev. Biophys. Bioeng.7, 497–526

    Google Scholar 

  • Anderson, R.G.W., Vasile, E., Mello, R.J., Brown, M.S., Goldstein, J.L. (1978) Immunocytochemical visualization of coated pits and vesicles in human fibroblasts: relation to low density lipoprotein receptor distribution. Cell15, 919–933

    Google Scholar 

  • Behnke, O. (1975) An outer component of microtubules. Nature (London)257, 709–710

    Google Scholar 

  • Bloodgood, R.A., Leffler, E.M., Bojczuk, A.T. (1979) Reversible inhibition ofChlamydomonas surface motility. J. Cell Biol.82, 664–674

    Google Scholar 

  • Brown, M.S., Goldstein, J.L. (1979) Receptor-mediated endocytosis: insights from the lipoprotein receptor system. Proc. Natl. Acad. Sci. USA76, 3330–3337

    Google Scholar 

  • Clarkson, D.T. (1977) Membrane structure and transport. In: Molecular biology of plant cells, pp. 24–63, Smith, H., ed. University of California Press, Berkeley California

    Google Scholar 

  • Cram, W.J. (1980) Pinocytosis in plants. New Phytologist84, 1–17

    Google Scholar 

  • Franke, W.W., Herth, W. (1974) Morphological evidence for de novo formation of coated vesicles in exponentially growing cultured plant cells. Exp. Cell. Res.89, 447–451

    Google Scholar 

  • Franke, W.W., Luder, M.R., Kartenbeck, J., Zerban, H., Keenan, T.W. (1976) Involvement of vesicle coat material in casein secretion and surface regeneration. J. Cell Biol.69, 173–195

    Google Scholar 

  • Giddings, T.H., Brower, D.L., Staehelin, L.A. (1980) Visualization of particle complexes in the plasma membrane ofMicrasterias denticulata associated with the formation of cellulose fibrils in primary and secondary cell walls. J. Cell Biol84, 327–339

    Google Scholar 

  • Goldstein, J.L., Anderson, R.G.W., Brown, M.S. (1979) Coated pits, coated vesicles and receptor-mediated endocytosis. Nature (London)279, 679–685

    Google Scholar 

  • Hardham, A.R., Gunning, B.E.S. (1978) Structure of cortical microtubule arrays in plant cells. J. Cell Biol.77, 14–34

    Google Scholar 

  • Hardham, A.R., Gunning, B.E.S. (1979) Interpolation of microtubules into cortical assays during cell elongation and differentiation in the roots ofAzolla pinnata. J. Cell Sci.37, 411–442

    Google Scholar 

  • Heath, I.B. (1974) Unified hypothesis for role of membrane-bound enzyme complexes and microtubules in plant-cell wall synthesis. J. Theor. Biol.48, 445–449

    Google Scholar 

  • Helenius, A., Kartenbeck, J., Simons, K., Fries, E. (1980) On the entry of Semliki Forest virus into BHK-21 cells. J. Cell Biol.84, 404–420

    Google Scholar 

  • Hepler, P.K., Palevitz, B.A. (2974) Microtubules and microfilaments. Annu. Rev. Plant Physiol.25, 309–362

    Google Scholar 

  • Heuser, J. (1980) Three-dimensional visualization of coated vesicle formation in fibroblasts. J. Cell Biol.84, 560–583

    Google Scholar 

  • Jacobs, M., Bennett, P.M., Dickens, M.J. (1975) Duplex microtubule is a new form of tubulin assembly induced by polycations. Nature (London)257, 707–709

    Google Scholar 

  • Keen, J.H., Willingham, M.C., Pastan, I.H. (1979) Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell16, 303–312

    Google Scholar 

  • Kirschner, M.W., Honig, L.S., Williams, R.C. (1975) Quantitative electron microscopy of microtubule assembly in vitro. J. Mol. Biol.99, 263–276

    Google Scholar 

  • Lloyd, C.W., Slabas, A.R., Powell, A.J., Lowe, S.B. (1980) Microtubules, protoplasts and cell shape. Planta140, 7–14

    Google Scholar 

  • Marchant, H.J., Hines, E.R. (1979) The role of microtubules and cell-wall deposition in elongation of regenerating protoplasts ofMougeotia. Planta146, 41–48

    Google Scholar 

  • Maxfield, F.R., Willingham, M.C., Schlessinger, J., Davies, P.J.A., Pastan, I. (1979) Receptor-mediated internalization of proteins and polypeptide hormones by cultured fibroblasts. In: Hormones and cell culture. Cold Spring Harbor conferences on cell proliferation, vol. 6, pp. 159–166. Cold Spring Harbor Press, Cold Spring Harbor, N.Y. USA

    Google Scholar 

  • Mazia, D., Mazia, G., Sale, Schatten and Sale, W. (1975) Adhesion of cells to surfaces coated with polylysine. J. Cell Biol.66, 198–200

    Google Scholar 

  • Newcomb, E.H. (1980) Coated vesicles: their occurrence in different plant cell types. In: Coated vesicles, pp. 55–68, Ockleford, C.D., Whyte, A., eds., Cambridge University Press, Cambridge, U.K.

    Google Scholar 

  • Nicolson, G.L. (1976) Cytoplasmic influence over cell surface components. Biophys. Biochim. Acta457, 57–108

    Google Scholar 

  • Olmsted, J.B., Marcum, J.M., Johnson, K.A., Allen, C., Borisy, G.G. (1974) Microtubule assembly: some possible regulatory mechanisms. J. Supramol. Struct.2, 429–450

    Google Scholar 

  • Palevitz, B.A. (1976) Actin cables and cytoplasmic streaming in green plants. In: Cell motility, Bk.C, pp. 601–611, Goldman, R., Pollard, T., Rosenbaum, J., eds. Cold Spring Harbor Press, Cold Spring Harbor, N.Y., USA

    Google Scholar 

  • Palevitz, B.A. (1980a) Comparative effects of phalloidin and cytochalasin B on motility and morphogenesis inAllium. Can. J. Bot.58, 773–785

    Google Scholar 

  • Palevitz, B.A. (1980b) The structure and development of stomatal cells. In: Soc. Exp. Biol. Sem. Ser., Stomatal Physiology, Jarvis, P.G., Mansfield, T.A., eds. Cambridge University Press, Cambridge, U.K., in press

    Google Scholar 

  • Palevitz, B.A., Hepler, P.K. (1976) Cellulose microfibril orientation and cell shaping in developing guard cells ofAllium—role of microtubules and ion accumulation. Planta132, 71–93

    Google Scholar 

  • Pearse, B.M.F. (1976) Clathrin: a unique protein associated with intracellular transfer of membrane by coated vesicles. Proc. Natl. Acad. Sci. USA73, 1255–1259

    Google Scholar 

  • Reynolds, E.S. (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol.17, 208–212

    Google Scholar 

  • Robinson, D.G. (1977) Plant cell wall synthesis. Adv. Bot. Res.5, 89–151

    Google Scholar 

  • Roth, T.F., Porter, K.R. (1964) Yolk protein uptake in the oocyte of the mosquitoAedes aegypti L. J. Cell Biol.20, 313–332

    Google Scholar 

  • Rothman, J.E., Fine, R.E. (1980) Coated vesicles transport newly synthesized membrane glycoproteins from endoplasmic reticulum to plasma membrane in two successive stages. Proc. Natl. Acad. Sci. USA77, 780–784

    Google Scholar 

  • Schnabl, H., Bornman, C.H., Ziegler, H. (1978) Studies on isolated starch-containing (Vicia faba) and starch-deficient (Allium cepa) guard cell protoplasts. Planta143, 33–39

    Google Scholar 

  • Seagull, R.W., Heath, I.B. (1979) The effects of tannic acid on the in vivo preservation of microfilaments. Eur. J. Cell Biol.20, 184–188

    Google Scholar 

  • Singh, A.P., Srivastava, L.M. (1973) The fine structure of pea stomata. Protoplasma76, 61–82

    Google Scholar 

  • Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res.26, 31–43

    Google Scholar 

  • Srivastava, L., Singh, A.P. (1972). Stomatal structure in corn leaves. J. Ultrastruct. Res.39, 345–363.

    Google Scholar 

  • Willingham, M.C., Maxfield, F.R., Pastan, I.H. (1979) 401-1 Macroglobulin binding to the plasma membrane of cultured fibroblasts. Diffuse binding followed by clustering in coated pits. J. Cell Biol.82, 614–625

    Google Scholar 

  • Woodward, M.P., Roth, T.F. (1978) Coated vesicles: characterization, selective dissociation and reassembly. Proc. Natl. Acad. Sci. USA75, 4394–4398

    Google Scholar 

  • Zeiger, E., Hepler, P.K. (1976) Production of guard cell protoplasts from onion and tobacco. Plant. Physiol.58, 492–498

    Google Scholar 

  • Zeiger, E., Hepler, P.K. (1977) Light and stomatal function: blue light stimulates swelling of guard cell protoplasts. Science196, 887–889

    Google Scholar 

  • Zeiger, E., Hepler, P.K. (1979) Blue light-induced, intrinsic vacuolar fluorescence in onion guard cells. J. Cell Sci.37, 1–10

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Doohan, M.E., Palevitz, B.A. Microtubules and coated vesicles in guard-cell protoplasts ofAllium cepa L.. Planta 149, 389–401 (1980). https://doi.org/10.1007/BF00571175

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00571175

Key words

Navigation