The effects of freezing on membrane electric potential in winter oilseed rape leaves

  • Grażyna Piotrowska
  • Maria Filek
  • Alina Kacperska


Extracellular ice formation in winter oilseed rape leaf discs (Brassica napus L. var. oleifera L. cv. Jantar) at different temperatures resulted in a transient membrane depolarization, which was followed by a decrease in membrane electric potential. In discs which underwent supercooling (no extracellular ice was formed), no membrane depolarization was observed. The inhibitors of calcium ion channels, gadolinium and lanthanum, decreased to some extend the amplitude of the frost-induced (−6 °C) depolarization and completely eliminated the decrease in membrane potential. Changes in membrane potential were associated with the increased electrolyte efflux, measured after thawing of the discs. No efflux from supercooled discs was observed. Application of calcium channel blockers decreased the level of the efflux induced by freezing at −6°C. It is suggested that membrane depolarization is one of the primary events induced by ice formation at a leaf surface. The possible reasons for changes in the membrane electric potential and their physiological consequences are discussed.

Key words

Brassica napus electrolyte efflux calcium channel inhibitors freezing gadolinium lanthanum leaves membrane potential winter oilseed rape 


  1. Arora, R., Palta, J.P. 1991. A loss of plasma membrane ATPase activity and its recovery coincides with incipient freeze-thaw injury and postthaw recovery in onion bulb scale tissue. Plant Physiol. 95: 846–852.PubMedGoogle Scholar
  2. Davies, E. 1987. Action potentials as multifunctional signals in plants: a unifying hypothesis to explain apperently disparate wound responses. Plant, Cell, Environ. 10: 623–631.CrossRefGoogle Scholar
  3. De Nisi, P., Zocchi, G. 1996. The role of calcium in the cold shock responses. Plant Sci. 121: 161–166.CrossRefGoogle Scholar
  4. Filek M., Pazurkiewicz-Kocot K., Dubert F., Marcińska I., Biesaga-Kościelniak J. 1993. Changes of surface potential and phospholipid composition of winter wheat callus cells grown at 5 and 25 C. J. Agronom. Crop Sci. 171: 243–250.CrossRefGoogle Scholar
  5. Frachisse, J.M., Desbiez, M.O., Champagnat, P., Thellier, M. 1985. Transmission of a traumatic signal via a wave of electric depolarization and induction of correlations between the cotyledonary buds in Bidens pilosus L. Physiol. Plant. 64: 48–52.CrossRefGoogle Scholar
  6. Hellegren, J., Widell, S., Lundborg, T. 1987. Freezing injury in purified plasma membranes from cold acclimated and non-acclimated needles of Pinus silvestris: Is the plasma membrane bound ion-stimulated ATPase the primary site of freezing injury? In: Plant Cold Hardiness, ed. by P.H. Li, Alan R. Liss Press, New York: 211–230.Google Scholar
  7. Iswari, S., Palta, J.P. 1989. Plasma membrane ATPase activity following reversible and nonreversible freezing injury. Plant Physiol. 90: 1088–1095.PubMedGoogle Scholar
  8. Jian, L.C., Sun, L.H., Dong, H.Z. 1982. Adaptive changes in ATPase activity in the cells of winter wheat seedlings during cold hardening. Plant Physiol. 70: 127–131.PubMedGoogle Scholar
  9. Julien, J.L., Frachisse J.M. 1992. Involvement of the proton pump and proton conductance change in the wave of depolarization induced by wounding in Bidens pilosa. Can. J. Bot. 70: 1451–1458.Google Scholar
  10. Kinoshita, T., Nishimura, M., Shimazaki, K. 1995. Cytosolic concentration of Ca2+ regulates the plasma membrane H+-ATPase in guard cells of fava bean. Plant Cell 7: 1333–1342.PubMedCrossRefGoogle Scholar
  11. Knight, M.R., Campbell, A.K., Smith, S.M., Trewavas, A.J. 1991. Transgenic plants aequorin reports the effects of touch and cold-shock, and elicitors on cytoplasmic calcium. Nature 352: 524–526.PubMedCrossRefGoogle Scholar
  12. Kulesza, L., Pukacki, P., Kacperska, A. 1986. Ice formation and frost killing temperatures related to cold acclimation of winter rape plants. Acta Physiol. Plant. 8: 185–193.Google Scholar
  13. Lewis, B.D. Karlin-Neumann G., Davis R.W., Spalding E.P. 1997. Ca2+-activated anion channels and membrane depolarizations induced by blue light and cold in Arabidopsis seedlings. Plant Physiol. 114: 1327–1334.PubMedCrossRefGoogle Scholar
  14. Lewis, B.D., Spalding E.P. 1998. Nonselective block by La3+ of Arabidopsis channels involved in signal transduction. J. Membrane Biol. 162: 81–90.CrossRefGoogle Scholar
  15. Millet, B., Pickard, B.G. 1988. Gadolinium ion is an inhibitor for testing the putative role of stretch-activated ion channels in geotropism and thigmotropism. Biophys. J. 53: 155a.Google Scholar
  16. Minorsky, P.V., Spanswick, R.M. 1989. Electro-physiological evidence for a role for calcium in temperature sensing by roots of cucumber seedlings. Plant, Cell, Environ. 12: 137–143.CrossRefGoogle Scholar
  17. Piňeros, M., Tester, M. 1997. Calcium channels in higher plant cells: selectivity, regulation and pharmacology. J. Exp. Bot. 48: 551–577.Google Scholar
  18. Piotrowska, G. 1988. The role of calcium ions in freezing injuries of winter oilseed rape leaves. Effects of calcium channel blockers and mimesis by calcium ionophore. Acta Physiol. Plant. 20: 257–261.Google Scholar
  19. Piotrowska, G., Kacperska, A. 1990. Utility of leaf discs cultured in vitro for studies on frost resistance. Plant Cell Tiss. Org. Cult. 22: 21–26.Google Scholar
  20. Ricca, U. 1916. Soluzione d’un problema di fisiologia: la propagazione di stimulo nella Mimosa. Nuovo G. Bot. Ital. 23: 51–170.Google Scholar
  21. Reid R.J., Tester, M., Smith, F.A. 1997. Voltage control of calcium influx in intact cells. Aust. J. Plant Physiol. 24: 805–810.CrossRefGoogle Scholar
  22. Sakai, A., Larcher, W. 1987. Frost Survival of Plants. Responses and Adaptation to Freezing Stress. Springer Verlag, Berlin, Heidelberg, New York: 21–36.Google Scholar
  23. Stanković, B., Zawadzki, T., Davies, E. 1997. Characterization of the variation potential in sunflower. Plant Physiol. 115: 1083–1088.PubMedGoogle Scholar
  24. Stahlberg, R., Cosgrove, D.J. 1992. Rapid alterations in growth rate and electrical potentials upon stem excision in pea seedlings. Planta 187: 523–531.PubMedCrossRefGoogle Scholar
  25. Stahlberg, R., Cosgrove, D.J. 1997. The propagation of slow wave potentials in pea epicotyls. Plant Physiol. 113: 209–217.PubMedGoogle Scholar
  26. Steponkus, P. Stout, D.G., Wolfe, J., Lovelase, R.V.E. 1985. Possible role of transient electric fields in freezing-induced membrane destabilization. J. Membrane Biol. 85: 191–198.CrossRefGoogle Scholar
  27. Smoleńska-Sym, G., Kacperska, A. 1996. Inositol 1,4,5-trisphosphate formation in leaves of winter oilseed rape plants in response to freezing, tissue water potential and abscisic acid. Physiol. Plant. 96: 692–698.CrossRefGoogle Scholar
  28. Sussman, M. 1993. The plasma membrane proton pump (H+-ATPase) of higher plants. In: Plant Signals in Interactions with Other Organisms, ed. by J. Schultz, I. Raskin, American Society of Plant Physiologists.Google Scholar
  29. Wayne, R. 1994. The excitability of plant cells: with a special emphasis on Characea internode cells. Bot. Rev. 60: 265–367.PubMedGoogle Scholar
  30. Wildon, D.C., Tain, J.F., Minchin, P.E.H., Gubb., I.R. Reilly, A.J., Skipper, Y.D., Doherty, H.M., O’Donnel P.J., Bowles, D.J. 1992. Electrical signaling and systemic proteinase inhibitor induction in the wounded plant. Nature 360: 62–65.CrossRefGoogle Scholar
  31. Yoshida, S. 1979. Freezing injury and phospholipid degradation in vivo in woody plant cells. II. Regulatory effects of divalent cations on activity of membrane-bound phospholipase D. Plant Physiol. 64: 247–273.PubMedCrossRefGoogle Scholar

Copyright information

© Department of Plant Physiology 2000

Authors and Affiliations

  • Grażyna Piotrowska
    • 1
  • Maria Filek
    • 2
  • Alina Kacperska
    • 1
  1. 1.Institute of Experimental Plant BiologyWarsaw UniversityWarszawa
  2. 2.F. Górski Department of Plant PhysiologyPolish Academy of SciencesKraków

Personalised recommendations