Skip to main content
SpringerLink
Log in

RGD-mediated membrane-matrix adhesion triggers agarose-induced embryoid formation in sunflower protoplasts

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Agarose embedding of sunflower (Helianthus annuus L.) hypocotyl protoplasts induces an asymmetric division pattern and subsequent polarized development leading to embryoid formation. We cultured protoplasts in media with different mannitol concentrations. Induction of plasmolysis of agarose-embedded protoplasts by increasing the mannitol concentration lowered the proportion of embryoids formed. This indicates that adhesion sites between the plasma membrane and the agarose matrix are involved in embryoid formation. The involvement of such adhesion sites was tested by incubating embedded protoplasts with RGD peptide. 1 μM RGD heptapeptide reduced embryoid formation by 50% as compared to the control DGR peptide. We also showed that RGD heptapeptide acts on the cytoskeleton by disrupting cortical microtubules. The results are discussed in terms of a model in which the anchorage of the protoplast membrane to the agarose matrix is mediated by RGD-binding proteins connected with microtubules, determining asymmetric division of the cell and polarized development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ECM:

extracellular matrix

TBS:

Tris-buffered saline

References

  • Asano Y, Ito Y, Sugiura K, Fujiie A (1994) Improved protoplast culture of bentgrass (Agrostis L.) using a medium with increased agarose concentration. J Plant Physiol 143: 122–124

    Google Scholar 

  • Bachewich C, Heath IB (1993) Effects of cell wall-cytoskeleton linkage inhibition on growth and development inSaprolegnia ferax. Inoculum 44: 25

    Google Scholar 

  • Barbotin JN, Nava Saucedo JE, Bazinet C, Kersulec A, Thomasset B, Thomas D (1993) Immobilization of whole cells and somatic embryos: coating process and cell-matrix interactions. In: Redenbaugh K (ed) Synseeds: application of synthetic seeds to crop improvement. CRC Press, Boca Raton, pp 65–103

    Google Scholar 

  • Barthou H, Brière C, Caumont C, Petitprez M, Kallerhoff J, Borin C, Souvré A, Alibert G (1997) Effect of atmospheric pressure on sunflower (Helianthus annuus L.) protoplast division. Plant Cell Rep 16: 310–314

    Google Scholar 

  • Boucaut JC, Darribère T, Poole TJ, Aoyama H, Yamada KM, Thiery JP (1984) Biologically active synthetic peptide as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. J Cell Biol 99: 1822–1830

    Google Scholar 

  • Caumont C, Petitprez M, Woynaroski S, Barthou H, Brière C, Kallerhoff J, Borin C, Souvré A, Alibert G (1997) Agarose embedding affects cell wall regeneration and microtubule organization in sunflower hypocotyl protoplasts. Physiol Plant 99: 129–134

    Google Scholar 

  • Chanabé C, Burrus M, Alibert G (1989) Factors affecting the improvement of colony formation from sunflower protoplasts. Plant Sci 64: 125–132

    Google Scholar 

  • — —, Bidney D, Alibert G (1991) Studies on plant regeneration from protoplasts in the genusHelianthus. Plant Cell Rep 9: 635–638

    Google Scholar 

  • Corrêa A, Staples RC, Hoch HC (1996) Inhibition of thigmostimulated cell differentiation with RGD-peptides inUromyces germlings. Protoplasma 194: 91–102

    Google Scholar 

  • Gens JS, Reuzeau C, Doolittle KW, McNally JG, Pickard BG (1996) Covisualization by computational optical-sectioning microscopy of integrin and associated proteins at the cell membrane of living onion protoplasts. Protoplasma 194: 215–230

    Google Scholar 

  • Haas TA, Plow EF (1994) Integrin-ligand interactions: a year in review. Curr Opin Cell Biol 6: 656–662

    Google Scholar 

  • Haubner R, Gratias R, Diefenbach B, Goodman SL, Jonczyk A, Kessler H (1996) Structural and functional aspects of RGD-containing cyclic pentapeptides as highly potent and selective integrin antagonists. J Am Chem Soc 118: 7461–7472

    Google Scholar 

  • Henry CA, Jordan JR, Kropf DL (1996) Localized membrane-wall adhesions inPelvetia zygotes. Protoplasma 190: 39–52

    Google Scholar 

  • Hynes RO (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69: 11–25

    Google Scholar 

  • Katembe WJ, Swatzell LJ, Makaroff CA, Kiss JZ (1997) Immunolocalization of integrin-like proteins inArabidopsis andChara. Physiol Plant 99: 7–14

    Google Scholar 

  • Klotz SA, Smith RL (1991) A fibronectin receptor onCandida albicans mediates adherence of the fungus to extracellular matrix. J Infect Dis 163: 604–610

    Google Scholar 

  • Lénée P, Chupeau Y (1986) Development of nitrogen assimilating enzymes during growth of cell derived from protoplasts of sunflower and tabacco. Plant Sci 59: 109–117

    Google Scholar 

  • Lloyd CW (1994) Why should stationary plant cells have such dynamic microtubules? Mol Biol Cell 5: 1277–1280

    Google Scholar 

  • Maniotis AJ, Chen CS, Ingber DE (1997) Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. Proc Natl Acad Sci USA 94: 849–854

    Google Scholar 

  • Novotny AM, Forman M (1974) The relationships between changes in cell wall composition and the establishment of polarity inFucus embryos. Dev Biol 40: 162–173

    Google Scholar 

  • O'Neill CM, Mathias RJ (1993) Regeneration of plants from protoplasts ofArabidopsis thaliana L. cv. Columbia (C24), via direct embryogenesis. J Exp Bot 44: 1579–1585

    Google Scholar 

  • Oparka KJ (1994) Plasmolysis: new insights into an old process. New Phytol 126: 571–591

    Google Scholar 

  • Pavalko FM, Otey CA (1994) Role of adhesion molecule cytoplasmic domains in mediating interactions with the cytoskeleton. Proc Soc Exp Biol Med 205: 282–293

    Google Scholar 

  • Petitprez M, Brière C, Borin C, Kallerhoff J, Souvré A, Alibert G (1995) Characterisation of protoplasts from hypocotyls ofHelianthus annuus in relation to their tissular origin. Plant Cell Tissue Organ Cult 41: 33–40

    Google Scholar 

  • Pont-Lezica RF, McNally JG, Pickard BG (1993) Wall-to-membrane linkers in onion epidermis: some hypotheses. Plant Cell Environ 16: 111–123

    Google Scholar 

  • Quatrano RS, Brian L, Aldridge J, Schultz T (1991) Polar axis fixation inFucus zygotes: components of the cytoskeleton and extracellular matrix. Development Suppl 1: 11–16

    Google Scholar 

  • Reuzeau C, Pont-Lezica RF (1995) Comparing plant and animal extracellular matrix—cytoskeleton connections — are they alike? Protoplasma 186: 113–121

    Google Scholar 

  • Robinson CR, Sligar SG (1995) Hydrostatic and osmotic pressure as tools to study macromolecular recognition. Methods Enzymol 259: 395–427

    Google Scholar 

  • Ryu JH, Mizuno K, Takagi S, Nagai R (1997) Extracellular components implicated in the stationnary organization of the actin cytoskeleton in mesophyll cells ofVallisneria. Plant Cell Physiol 38: 420–432

    Google Scholar 

  • Shibaoka H (1993) Regulation by gibberellins of the orientation of cortical microtubules in plant cells. Aust J Plant Physiol 20: 461–470

    Google Scholar 

  • Schindler M, Meiners S, Cheresh DA (1989) RGD-dependent linkage between plant cell wall and plasma membrane: consequences for growth. J Cell Biol 108: 1955–1965

    Google Scholar 

  • Schnabl H, Youngman RJ (1985) Immobilisation of plant cell protoplasts inhibits enzymic lipid peroxidation. Plant Sci 40: 65–69

    Google Scholar 

  • Smidsrod O, Skjak-Braek G (1990) Alginate as immobilization matrix for cells. Trends Biotechnol 8: 71–78

    Google Scholar 

  • Traas J (1989) Le rôle du cytosquelette dans la détermination du plan de division. Bull Soc Bot Fr 136: 99–106

    Google Scholar 

  • Wayne R, Staves MP, Leopold AC (1992) The contribution of the extracellular matrix to gravisensing in characean cell. J Cell Sci 101: 611–623

    Google Scholar 

  • Woldringh CL (1994) Significance of plasmolysis spaces as markers for periseptal annuli and adhesion sites. Mol Microbiol 14: 597–607

    Google Scholar 

  • Wyatt SE, Carpita NC (1993) The plant cytoskeleton-cell-wall continuum. Trends Cell Biol 3: 413–417

    Google Scholar 

  • Yuan M, Shaw PJ, Warn RM, Lloyd CW (1994) Dynamic reorientation of cortical microtubules from transverse to longitudinal in living plant cells. Proc Nat Acad Sci USA 91: 6050–6053

    Google Scholar 

  • Zandomeni K, Schopfer P (1994) Mechanosensory microtubule reorientation in the epidermis of maize coleoptiles subjected to bending stress. Protoplasma 182: 96–101

    Google Scholar 

  • Zhu JK, Shi J, Singh U, Wyatt SE, Bressan RA, Hasegawa PM, Carpita NC (1993) Enrichment of vitronectin- and fibronectinlike proteins in NaCl-adapted plant cells and evidence for their involvement in plasma membrane-cell wall adhesion. Plant J 3: 637–646

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Biotechnologie et Amélioration des Plantes, Ecole Nationale Supérieure Agronomique, Avenue de l'Agrobiopole, F-31326, Castanet-Tolosan Cedex, France

    H. Barthou, M. Petitprez, C. Brière, A. Souvré & G. Alibert

Authors
  1. H. Barthou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Petitprez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Brière
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. Souvré
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G. Alibert
    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

Barthou, H., Petitprez, M., Brière, C. et al. RGD-mediated membrane-matrix adhesion triggers agarose-induced embryoid formation in sunflower protoplasts. Protoplasma 206, 143–151 (1999). https://doi.org/10.1007/BF01279261

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01279261

Keywords

Search

Navigation