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Planta

, Volume 183, Issue 4, pp 590–596 | Cite as

Vanadate inhibition of stomatal opening in epidermal peels of Commelina communis

Cl interferes with vanadate uptake
  • Amnon Schwartz
  • Nitza Illan
  • Sarah M. Assmann
Article

Abstract

An H+ ATPase at the plasma-membrane of guard cells is thought to establish an electrochemical gradient that drives K+ and Cl uptake, resulting in osmotic swelling of the guard cells and stomatal opening. There are, however, conflicting results regarding the effectiveness of the plasma-membrane H+-ATPase inhibitor, vanadate, in inhibiting both H+ extrusion from guard cells and stomatal opening. We found that 1 mM vanadate inhibited light-stimulated stomatal opening in epidermal peels of Commelina communis L. only at KCl concentrations lower than 50 mM. When impermeant n-methylglucamine and HCl (pH 7.2) were substituted for KCl, vanadate inhibition was still not observed at total salt concentrations≥50 mM. In contrast, in the absence of Cl, when V2O5 was used to buffer KOH, vanadate inhibition of stomatal opening occurred at K+ concentrations as high as 70 mM. Partial vanadate inhibition was observed in the presence of the impermeant anion, iminodiacetic acid (100 mM KHN(CH2CO2H)2). These results indicate that high concentrations of permeant anions prevent vanadate uptake and consequently prevent its inhibitory effect. In support of this hypothesis, an inhibitor of anion uptake, anthracene-9-carboxylic acid, partially prevented vanadate inhibition of stomatal opening. Other anion-uptake inhibitors (1 mM 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid, 1 mM 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid, 200 μM Zn2+) were not effective. Decreased vanadate inhibition at high Cl/vanadate ratios may result from competition between vanadate and Cl for uptake. Unlike metabolic inhibitors, vanadate did not affect the extent of stomatal closure stimulated by darkness, further indicating that the observed action of vanadate represents a specific inhibition of the guard-cell H+ ATPase.

Key words

Anion transport Commelina Fusicoccin Guard cell Stomatal opening Vanadate, Vanadium 

Abbreviations

DIDS

4,4-diisothiocyanatostilbene-2,2′-disulfonic acid

FC

fusicoccin

SITS

4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid

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References

  1. Assmann, S.M., Zeiger, E. (1987) Guard cell bioenergetics. In: Stomatal function, pp. 165–194, Zeiger, E., Farquhar, G.D., Cowan, I., eds. Stanford University Press, Stanford, Cal., USAGoogle Scholar
  2. Assmann, S.M., Simoncini, L., Schroeder, J.I. (1985) Blue light activates electrogenic ion pumping in guard cell protoplasts of Vicia faba L. Nature 318, 285–287Google Scholar
  3. Blum, W., Key, G., Weiler, E.W. (1988) ATPase activity in plasmalemma-rich vesicles isolated by aqueous two-phase partitioning from Vicia faba mesophyll and epidermis: characterization and influence of abscisic acid and fusicoccin. Physiol Plant. 72, 279–287Google Scholar
  4. Bosma, M.M. (1989) Anion channels with multiple conductance levels in a mouse B lymphocyte cell line. J. Physiol. 410, 67–90Google Scholar
  5. Bowman, B.J., Slayman, C.W. (1979) The effect of vanadate on the plasma membrane ATPase of Neurospora crassa J. Biol. Chem. 254, 2928–2934Google Scholar
  6. Briskin, D.P., Leonard, R.T. (1982a) Partial characterization of a phosphorylated intermediate associated with the plasma membrane ATPase of corn roots. Proc. Natl. Acad. Sci. USA 79, 6922–6926Google Scholar
  7. Briskin, D.P., Leonard, R.T. (1982b) Phosphorylation of the adenosine triphosphatase in a deoxycholate-treated plasma membrane fraction from corn roots. Plant Physiol. 70, 1459–1464Google Scholar
  8. Cantley, L.C., Jr., Josephson, L., Warner, R., Vanagisann, M., Lechere C. Guidotti G. (1977) Vanadate is a potent (Na, K)-ATPase inhibitor found in ATP derived from muscle. J. Biol. Chem. 252, 7421–7423Google Scholar
  9. Cantley, L.C., Jr., Resh, M.D., Guidotti, G. (1978) Vanadate inhibits the red cell (Na+, K+) ATPase from the cytoplasmic side. Nature 272, 552–554Google Scholar
  10. Clint, G., Blatt, M.R. (1989) Mechanisms of fusicoccin action: evidence for concerted modulations of secondary K+ transport in a higher plant cell. Planta 178, 495–508Google Scholar
  11. Cocucci, M., Ballarin-Denti, A., Marré, M.I. (1980) Effects of orthovanadate on H+ secretion, K+ uptake, electric potential difference and membrane ATPase activities of higher plant tissue. Plant Sci. Lett. 17, 391–400Google Scholar
  12. Dhugga, K.S., Waines, J.G., Leonard, R.T. (1988) Nitrate absorption by corn roots: inhibition by phenylglyoxal. Plant Physiol. 86, 759–763Google Scholar
  13. Fricker, M.D., Wilmer, C.M. (1987) Vanadate sensitive ATPase and phosphate activity in guard cell protooplasts of Commelina. J. Exp. Bot. 38, 642–648Google Scholar
  14. Gallagher, S.R., Leonard, R.T. (1982) Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activities of corn roots. Plant Physiol. 70, 1335–1340Google Scholar
  15. Gepstein, S., Jacobs, M., Taiz, L. (1982) Inhibition of stomatal opening in Vicia faba epidermal tissue by vanadate and abscisic acid. Plant Sci. Lett. 28, 63–72Google Scholar
  16. Jacobs, M., Taiz, L. (1980) Vanadate inhibition of auxin-enhanced H+ secretion and elongation in pea epicotyls and oat coleoptiles. Proc. Natl. Acad. Sci. USA 77, 7242–7246Google Scholar
  17. Karlsson, P.E., Schwartz, A. (1988) Characterization of the effects of metabolic inhibitors, ATPase inhibitors, and a potassium-channel blocker on stomatal opening and closing in isolated epidermis of Commelina communis L. Plant Cell Environ. 11, 165–172Google Scholar
  18. Keller, B.U., Hedrich, R. Raschke, K. (1989) Voltage-dependent anion channels in the plasma membrane of guard cells. Nature 341, 450–453Google Scholar
  19. Kochian, L.V., Xin-zhi, J., Lucas, W.J. (1985) Potassium transport in corn roots. IV: Characterization of the linear component. Plant Physiol. 79, 771–776Google Scholar
  20. Kuroda, H., Warncke, J., Sanders, D., Hansen, U.P., Allen, K.E., Bowman, B.J. (1980) Effects of vanadate on the electrogenic proton pump in Neurospora, In: Plant membrane transport: Current conceptual issues, pp. 507–508, Spanswick, R.M., Lucas, W.J., Dainty, J., eds. Elsevier, AmsterdamGoogle Scholar
  21. Lin, W (1981) Inhibition of anion transport in corn root protoplasts. Plant Physiol. 68, 435–438Google Scholar
  22. MacRobbie, E.A.C. (1987) Ion relations of guard cells. In: Stomatal function, pp. 125–162, Zeiger, E., Farquhar, G.D., Cowan, I., eds. Stanford University Press, Stanford, Cal., USAGoogle Scholar
  23. Nobel, P.S. (1983) Biophysical plant physiology and ecology. W.H. Freeman & Co., San FranciscoGoogle Scholar
  24. Palade, P.T., Barchi, R.L. (1977) On the inhibition of muscle membrane chloride conductance by aromatic carboxylic acids. J. Gen. Physiol. 69, 879–896Google Scholar
  25. Pedersen, P.L., Carafoli, E. (1987) Ion motive ATPases: I. Ubiquity, properties, and significance to cell function. TIBS 12, 146–150Google Scholar
  26. Penny, M.G., Kelday, L.S., Bowling, D.J.F. (1976) Active chloride transport in the leaf epidermis of Commelina communis in relation to stomatal activity. Planta 130, 291–294Google Scholar
  27. Pope, M.T., Dale, B.W. (1969) Isopoly-vanadates, -niobates, and -tantalates. Q. Rev. Chem. Soc. Lond. 22, 527–548Google Scholar
  28. Raschke, K. (1987) Action of abscisic acid on guard cells. In: Stomatal function, pp. 254–279, Zeiger, E., Farquhar, G.D., Cowan, I., eds. Stanford University Press, Stanford, Cal., USAGoogle Scholar
  29. Raschke, K., Humble, G.D. (1973) No uptake of anions required by opening stomata of Vicia faba: guard cells release hydrogen ions. Planta 115, 47–57Google Scholar
  30. Raschke, K., Schnabl, H. (1978) Availability of chloride affects the balance between potassium chloride and potassium malate in guard cell of Vicia faba L. Plant Physiol. 62, 84–87Google Scholar
  31. Rubinson, K.A. (1981) Concerning the form of biochemically active vanadate. Proc. R. Soc. London Ser. B 212, 65–84Google Scholar
  32. Saxe, H., Rajagopal, R. (1981) Effect of vanadate on bean leaf movement, stomatal conductance, barley leaf unrolling, respiration, and phosphatase activity. Plant Physiol. 68, 880–884Google Scholar
  33. Schroeder, J.I., Hagiwara, S. (1989) Cytosolic calcium regulates ion channels in the plasma membrane of Vicia faba guard cells. Nature 338, 427–430Google Scholar
  34. Serrano, R. (1989) Structure and function of plasma membrane ATPase. Annu. Rev. Plant Physiol. Plant Mol. Biol. 40, 61–94Google Scholar
  35. Shimazaki, K., Kondo, N. (1987) Plasma membrane H+ -ATPase in guard-cell protoplasts from Vicia faba L. Plant Cell Physiol. 28, 893–900Google Scholar
  36. Shimazaki K., Iiono, M., Zeiger, E., (1986) Blue light-dependent proton extrusion by guard cell protoplasts of Vicia faba. Nature 319, 324–326Google Scholar
  37. Stein, W.D. (1986) Transport and diffusion across cell membranes. Academic Press, LondonGoogle Scholar
  38. Sze, H. (1984) H+-translocating ATPases of the plasma membrane and tonoplast of plant cells. Physiol. Plant. 61, 683–691Google Scholar
  39. Weyers, J.D.B., Patterson, N.W., Fitzsimmons, P.J., Dudley, J.M. (1982) Metabolic inhibitors block ABA-induced stomatal closure. J. Exp. Bot. 33, 1270–1278Google Scholar
  40. Zeiger, E. (1983) The biology of stomatal guard cells. Annu. Rev. Plant Physiol. 34, 441–475Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Amnon Schwartz
    • 1
  • Nitza Illan
    • 1
  • Sarah M. Assmann
    • 2
  1. 1.Department of Agricultural Botany, Faculty of AgricultureHebrew University of JerusalemRehovotIsrael
  2. 2.Department of Organismic and Evolutionary BiologyHarvard University, Biological LaboratoriesCambridgeUSA

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