Skip to main content

Organization of the actin cytoskeleton during pollen development inGasteria verrucosa (Mill.) H. Duval visualized with rhodamine-phalloidin

Planta volume 178, pages 531–539 (1989)Cite this article

Abstract

The three-dimensional organization of the microfilamental cytoskeleton of developingGasteria pollen was investigated by light microscopy using whole cells and fluorescently labelled phalloidin. Cells were not fixed chemically but their walls were permeabilized with dimethylsulphoxide and Nonidet P-40 at premicrospore stages or with dimethylsulphoxide, Nonidet P-40 and 4-methylmorpholinoxide-monohydrate at free-microspore and pollen stages to dissolve the intine.

Four strikingly different microfilamentous configurations were distinguished. (i) Actin filaments were observed in the central cytoplasm throughout the successive stages of pollen development. The network was commonly composed of thin bundles ramifying throughout the cytoplasm at interphase stages but as thick bundles encaging the nucleus prior to the first and second meiotic division. (ii) In released microspores and pollen, F-actin filaments formed remarkably parallel arrays in the peripheral cytoplasm. (iii) In the first and second meiotic spindles there was an apparent localization of massive arrays of phalloidin-reactive material. Fluorescently labelled F-actin was present in kinetochore fibers and pole-to-pole fibers during metaphase and anaphase. (iv) At telophase, microfilaments radiated from the nuclear envelopes and after karyokinesis in the second meiotic division, F-actin was observed in phragmoplasts.

We did not observe rhodamine-phalloidin-labelled filaments in the cytoplasm after cytochalasin-B treatment whereas F-actin persisted in the spindle. Incubation at 4° C did not influence the existence of cytoplasmic microfilaments whereas spindle filaments disappeared. This points to a close interdependence of spindle microfilaments and spindle tubules.

Based on present data and earlier observations on the configuration of microtubules during pollen development in the same species (Van Lammeren et al., 1985, Planta165, 1-11) there appear to be apparent codistributions of F-actin and microtubules during various stages of male meiosis inGasteria verrucosa.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Abbreviations

DMSO:

dimethylsulfoxide

References

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

    Google Scholar 

  • Baldi, B.G., Franceschi, V.R., Loewus, F.A. (1986) Dissolution of pollen intine and release of sporoplasts. In: Biotechnology and ecology of pollen, pp. 77–82, Mulcahy, D.L., Mulcahy, G.B., Ottaviano, E., eds. Springer, New York Berlin Heidelberg Tokyo

    Google Scholar 

  • Clayton, L., Lloyd, C.W. (1985) Actin organization during the cell cycle in meristematic plant cells. Exp. Cell Res.156, 231–238

    Google Scholar 

  • Condeelis, J.S. (1974) The identification of F-actin in the pollen tube and protoplast ofAmaryllis belladonna. Exp. Cell Res.88, 435–439

    Google Scholar 

  • Forer, A., Jackson, W.T. (1979) Actin in spindles ofHaemanthus katherinae endosperm. I. General results using various glycerination methods. J. Cell Sci.37, 323–347

    Google Scholar 

  • Heath, I.B., Seagull, R.W. (1982) Oriented cellulose fibrils and the cytoskeleton: a critical comparison of models. In: The cytoskeleton in plant growth and development, pp. 163–187, Lloyd, C.W., ed. Academic Press, London

    Google Scholar 

  • Herth, W., Franke, W.W., Vanderwoude, W.J. (1972) Cytochalasin stops tip growth in plants. Naturwissenschaften59, 38–39

    Google Scholar 

  • Heslop-Harrison, J., Heslop-Harrison, Y., Cresti, M., Tiezzi, A., Chiampolini, F. (1986) Actin during pollen germination. J. Cell Sci.86, 1–8

    Google Scholar 

  • Jackson, W.T. (1982) Actomyosin. In: The cytoskeleton in plant growth and development, pp. 3–29, Lloyd, C.W., ed. Academic Press, London

    Google Scholar 

  • Kakimoto, T., Shibaoka, H. (1987) Actin filaments and microtubules in the preprophase band and phragmoplast of tobacco cells. Protoplasma140, 151–156

    Google Scholar 

  • Lin, D.C., Tobin, K.D., Grumet, M., Lin, S. (1980) Cytochalasins inhibit nuclei-induced actin polymerization by blocking filament elongation. J. Cell Biol.84, 455–460

    Google Scholar 

  • Lloyd, C.W., ed. (1982) The cytoskeleton in plant growth and development. Academic Press, London

    Google Scholar 

  • Lloyd, C.W. (1987) The plant cytoskeleton: The impact of fluorescence microscopy. Annu. Rev. Plant Physiol.38, 119–139

    Google Scholar 

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

    Google Scholar 

  • Palevitz, B.A. (1987) Actin in the preprophase band ofAllium cepd. J. Cell Biol.104, 1515–1519

    Google Scholar 

  • Parthasarathy, M.V., Perdue, T.D., Witztum, A., Alvernaz, J. (1985) Actin network as a normal component of the cytoskeleton in many vascular plant cells. Am. J. Bot.72, 1318–1323

    Google Scholar 

  • Pesacreta, T.C., Carley, W.W., Webb, W.W., Parthasarathy, M.V. (1982) F-actin in conifer roots. Proc. Natl. Acad. Sci. USA79, 2898–2901

    Google Scholar 

  • Pierson, E.S. (1988) Rhodamine-phalloidin staining of F-actin in pollen after dimethylsulphoxide permeabilization. A comparison with the conventional formaldehyde preparation. Sex. Plant Reprod.1, 83–87

    Google Scholar 

  • Pierson, E.S., Derksen, J., Traas, J.A. (1986) Organization of microfilaments and microtubules in pollen tubes grown in vitro or in vivo in various angiosperms. Eur. J. Cell Biol.41, 14–18

    Google Scholar 

  • Schliwa, M. ed. (1986) The cytoskeleton, an introductory survey. Springer, Wien New York

    Google Scholar 

  • Schmit, A.C., Lambert, A.M. (1985) F-actin distribution during the cell cycle of higher plant endosperm cells. J. Cell Biol.101, 38a

    Google Scholar 

  • Schmit, A.C., Lambert, A.M. (1987) Characterization and dynamics of cytoplasmic F-actin in higher plant endosperm cells during interphase, mitosis and cytokinesis. J. Cell Biol.105, 2157–2166

    Google Scholar 

  • Schmit, A.C., Vantard, M., Lambert, A.M. (1985) Microtubule and F-actin rearrangement during the initiation of mitosis in acentriolar higher plant cells. In: Cell motility: mechanism and regulation, pp. 415–433, Ishikawa H., Hatano, S., Sato, H., eds. University of Tokyo Press

  • Seagull, R., Falconer, M., Weerdenburg, C. (1987) Microfilaments: Dynamic arrays in higher plant cells. J. Cell Biol.104, 995–1004

    Google Scholar 

  • Sheldon, J.M., Hawes, C. (1988) The actin cytoskeleton during male meiosis inLilium. Cell Biol. Int. Rep.12, 471–476

    Google Scholar 

  • Staiger, C.J., Schliwa, M. (1987) Actin localization and function in higher plants. Protoplasma141, 1–12

    Google Scholar 

  • Tiwari, S., Wick, S.M., Williamson, R.E., Gunning, B.E.S. (1984) Cytoskeleton and integration of cellular function in cells of higher plants. J. Cell Biol.99, 63s-69s

    Google Scholar 

  • Traas, J.A., Doonan, J.H., Rawlins, D.J., Shaw, P.J., Watts, J., Lloyd, C.W. (1987) An actin network is present in the cytoplasm throughout the cell cycle of carrot cells and associates with the dividing nucleus. J. Cell Biol.105, 387–395

    Google Scholar 

  • Van Lammeren, A.A.M., Keijzer, C.J., Willemse, M.T.M., Kieft, H. (1985) Structure and function of the microtubular cytoskeleton during pollen development inGasteria verrucosa (Mill.) H. Duval. Planta165, 1–11

    Google Scholar 

  • Wieland, T. (1977) Modifications of actin by phallotoxins. Naturwissenschaften64, 303–309

    Google Scholar 

  • Willemse, M.Th.M. (1972) Morphological and quantitative changes in the population of cell organelles during microsporogenesis ofGasteria verrucosa. Acta Bot. Neerl.21, 17–31

    Google Scholar 

  • Wulf, E., Deboben, A., Bautz, F.A., Faulstich, H., Wieland, T. (1979) Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc. Natl. Acad. Sci. USA76, 4498–4502

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Cytology and Morphology, Agricultural University Wageningen, Arboretumlaan 4, NL-6703 BD, Wageningen, The Netherlands

    A. A. M. Van Lammeren & M. T. M. Willemse

  2. Plant Anatomy and Cytology Department, Maria Curie Slodowska University Lublin, Akademicka 19, 2033, Lublin, Poland

    J. Bednara

Authors
  1. A. A. M. Van Lammeren
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Bednara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. T. M. Willemse
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Van Lammeren, A.A.M., Bednara, J. & Willemse, M.T.M. Organization of the actin cytoskeleton during pollen development inGasteria verrucosa (Mill.) H. Duval visualized with rhodamine-phalloidin. Planta 178, 531–539 (1989). https://doi.org/10.1007/BF00963823

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00963823

Key words

  • Cytoskeleton
  • Gasteria
  • Meiosis
  • Microfilament
  • Pollen
  • Rhodamine-phalloidin