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.
Similar content being viewed by others
Abbreviations
- DIDS:
-
4,4-diisothiocyanatostilbene-2,2′-disulfonic acid
- FC:
-
fusicoccin
- SITS:
-
4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid
References
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., USA
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, 285–287
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, 279–287
Bosma, M.M. (1989) Anion channels with multiple conductance levels in a mouse B lymphocyte cell line. J. Physiol. 410, 67–90
Bowman, B.J., Slayman, C.W. (1979) The effect of vanadate on the plasma membrane ATPase of Neurospora crassa J. Biol. Chem. 254, 2928–2934
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, 6922–6926
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, 1459–1464
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, 7421–7423
Cantley, L.C., Jr., Resh, M.D., Guidotti, G. (1978) Vanadate inhibits the red cell (Na+, K+) ATPase from the cytoplasmic side. Nature 272, 552–554
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, 495–508
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, 391–400
Dhugga, K.S., Waines, J.G., Leonard, R.T. (1988) Nitrate absorption by corn roots: inhibition by phenylglyoxal. Plant Physiol. 86, 759–763
Fricker, M.D., Wilmer, C.M. (1987) Vanadate sensitive ATPase and phosphate activity in guard cell protooplasts of Commelina. J. Exp. Bot. 38, 642–648
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, 1335–1340
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, 63–72
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, 7242–7246
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, 165–172
Keller, B.U., Hedrich, R. Raschke, K. (1989) Voltage-dependent anion channels in the plasma membrane of guard cells. Nature 341, 450–453
Kochian, L.V., Xin-zhi, J., Lucas, W.J. (1985) Potassium transport in corn roots. IV: Characterization of the linear component. Plant Physiol. 79, 771–776
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, Amsterdam
Lin, W (1981) Inhibition of anion transport in corn root protoplasts. Plant Physiol. 68, 435–438
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., USA
Nobel, P.S. (1983) Biophysical plant physiology and ecology. W.H. Freeman & Co., San Francisco
Palade, P.T., Barchi, R.L. (1977) On the inhibition of muscle membrane chloride conductance by aromatic carboxylic acids. J. Gen. Physiol. 69, 879–896
Pedersen, P.L., Carafoli, E. (1987) Ion motive ATPases: I. Ubiquity, properties, and significance to cell function. TIBS 12, 146–150
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, 291–294
Pope, M.T., Dale, B.W. (1969) Isopoly-vanadates, -niobates, and -tantalates. Q. Rev. Chem. Soc. Lond. 22, 527–548
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., USA
Raschke, K., Humble, G.D. (1973) No uptake of anions required by opening stomata of Vicia faba: guard cells release hydrogen ions. Planta 115, 47–57
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, 84–87
Rubinson, K.A. (1981) Concerning the form of biochemically active vanadate. Proc. R. Soc. London Ser. B 212, 65–84
Saxe, H., Rajagopal, R. (1981) Effect of vanadate on bean leaf movement, stomatal conductance, barley leaf unrolling, respiration, and phosphatase activity. Plant Physiol. 68, 880–884
Schroeder, J.I., Hagiwara, S. (1989) Cytosolic calcium regulates ion channels in the plasma membrane of Vicia faba guard cells. Nature 338, 427–430
Serrano, R. (1989) Structure and function of plasma membrane ATPase. Annu. Rev. Plant Physiol. Plant Mol. Biol. 40, 61–94
Shimazaki, K., Kondo, N. (1987) Plasma membrane H+ -ATPase in guard-cell protoplasts from Vicia faba L. Plant Cell Physiol. 28, 893–900
Shimazaki K., Iiono, M., Zeiger, E., (1986) Blue light-dependent proton extrusion by guard cell protoplasts of Vicia faba. Nature 319, 324–326
Stein, W.D. (1986) Transport and diffusion across cell membranes. Academic Press, London
Sze, H. (1984) H+-translocating ATPases of the plasma membrane and tonoplast of plant cells. Physiol. Plant. 61, 683–691
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, 1270–1278
Zeiger, E. (1983) The biology of stomatal guard cells. Annu. Rev. Plant Physiol. 34, 441–475
Author information
Authors and Affiliations
Additional information
We thank Drs. R.T. Leonard (University of California, Riverside, USA) and K.A, Rubinson (Yellow Springs, Oh., USA) for helpful comments on the research, Janet Sherwood (Harvard University) for excellent plant care, and Angela Ciamarra, Anne Gershenson, Gustavo Lara (Harvard University) and Orit Tal (Hebrew University) for valuable technical assistance. This research was supported by a grant from the National Science Foundation (DCB-8904041) to S.M.A.
Rights and permissions
About this article
Cite this article
Schwartz, A., Illan, N. & Assmann, S.M. Vanadate inhibition of stomatal opening in epidermal peels of Commelina communis . Planta 183, 590–596 (1991). https://doi.org/10.1007/BF00194281
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00194281