Ion Channels in the Xylem Parenchyma of Barley Roots (A Procedure to Isolate Protoplasts from This Tissue and a Patch-Clamp Exploration of Salt Passageways into Xylem Vessels

To identify mechanisms for the simultaneous release of anions and cations into the xylem sap in roots, we investigated voltage-dependent ion conductances in the plasmalemma of xylem parenchyma cells. We applied the patch-clamp technique to protoplasts isolated from the xylem parenchyma by differential enzymic digestion of steles of barley roots (Hordeum vulgare L. cv Apex). In the whole-cell configuration, three types of cation-selective rectifiers could be identified: (a) one activated at membrane potentials above about -50 mV; (b) a second type of outward current appeared at membrane potentials above +20 to +40 mV; (c) below a membrane potential of approximately -110 mV, an inward rectifier could be distinguished. In addition, an anion-specific conductance manifested itself in single-channel activity in a voltage range extending from about -100 to +30 mV, with remarkably slow gating. In excised patches, K+ channels activated at hyperpolarization as well as at depolarization. We suggest that salt is released from the xylem parenchyma into the xylem apoplast by simultaneous flow of cations and anions through channels, following electrochemical gradients set up by the ion uptake processes in the cortex and, possibly, the release and reabsorption of ions on their way to the xylem.

[1]  R. Hedrich,et al.  Ca2+ and nucleotide dependent regulation of voltage dependent anion channels in the plasma membrane of guard cells. , 1990 .

[2]  D. Clarkson,et al.  A Comparison Between the ATPase and Proton Pumping Activities of Plasma Membranes Isolated from the Stele and Cortex of Zea mays Roots , 1993 .

[3]  D. Clarkson,et al.  Proton Fluxes and the Activity of a Stelar Proton Pump in Onion Roots , 1986 .

[4]  J. Dunlop,et al.  The Movement of Ions to the Xylem Exudate of Maize Roots I. PROFILES OF MEMBRANE POTENTIAL AND VACUOLAR POTASSIUM ACTIVITY ACROSS THE ROOT , 1971 .

[5]  C. Slayman,et al.  Ion channels in Arabidopsis plasma membrane : transport characteristics and involvement in light-induced voltage changes. , 1992, Plant physiology.

[6]  S. Tyerman,et al.  Current-Voltage Curves of Single CI− Channels which Coexist with Two Types of K+ Channel in the Tonoplast of Chara corallina , 1989 .

[7]  K. Ketchum,et al.  Characterization of potassium-dependent currents in protoplasts of corn suspension cells. , 1989, Plant physiology.

[8]  E. Neher Correction for liquid junction potentials in patch clamp experiments. , 1992, Methods in enzymology.

[9]  K. Takeda,et al.  Calcium-activated, voltage-dependent, non-selective cation currents in endosperm plasma membrane from higher plants , 1989, Proceedings of the Royal Society of London. B. Biological Sciences.

[10]  R. Hedrich The Physiology of ION Channels and Electrogenic Pumps in Higher Plants , 1989 .

[11]  E Marre,et al.  Fusicoccin: A Tool in Plant Physiology , 1979 .

[12]  A. Crafts,et al.  MIGRATION OF SALTS AND WATER INTO XYLEM OF THE ROOTS OF HIGHER PLANTS , 1938 .

[13]  M. Goldsmith,et al.  K Channels Are Responsible for an Inwardly Rectifying Current in the Plasma Membrane of Mesophyll Protoplasts of Avena sativa. , 1992, Plant physiology.

[14]  R. Graham,et al.  A Micro-electrode Fast Data Capture Technique: Its Use to Examine Plant Cell Electrophysiology in the Wheat Root , 1987 .

[15]  A. Spurr,et al.  Lateral Transport of Ions into the Xylem of Corn Roots: II. Evaluation of a Stelar Pump. , 1971, Plant physiology.

[16]  K. Raschke,et al.  A slow anion channel in guard cells, activating at large hyperpolarization, may be principal for stomatal closing , 1992, FEBS letters.

[17]  M. G. Pitman,et al.  ION Transport into the Xylem , 1977 .

[18]  D. Ypey,et al.  Whole-Cell K+ Currents across the Plasma Membrane of Tobacco Protoplasts from Cell-Suspension Cultures , 1993, Plant physiology.

[19]  G. Ehrenstein,et al.  Potassium Channels in Motor Cells of Samanea saman: A Patch-Clamp Study. , 1988, Plant physiology.

[20]  J. B. Hanson Application of the chemiosmotic hypothesis to ion transport across the root. , 1978, Plant physiology.

[21]  D. Kramer The possible role of transfer cells in the adaptation of plants to salinity , 1983 .

[22]  A. D. Boer Xylem/Symplast ion exchange : Mechanism and function in salt-tolerance and growth , 1985 .

[23]  K Raschke,et al.  Voltage dependence of K channels in guard-cell protoplasts. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[24]  J. Dunlop Membrane Potentials in the Xylem in Roots of Intact Plants , 1982 .

[25]  A. D. Boer,et al.  Xylem perfusion of tap root segments of Plantago maritima: the physiological significance of electrogenic xylem pumps , 1985 .

[26]  R. D. Warmbrodt STUDIES ON THE ROOT OF HORDEUM VULGARE L.– ULTRASTRUCTURE OF THE SEMINAL ROOT WITH SPECIAL REFERENCE TO THE PHLOEM , 1985 .

[27]  R. Cerana,et al.  K+ and Cl− Conductance of Arabidopsis thaliana Plasma Membrane at Depolarized Voltages , 1992 .

[28]  M. Tester Tansley Review No. 21 Plant ion channels: whole-cell and single channel studies. , 1990, The New phytologist.

[29]  Klaus Raschke,et al.  Voltage-dependent anion channels in the plasma membrane of guard cells , 1989, Nature.

[30]  K. Föhr,et al.  Calculation and control of free divalent cations in solutions used for membrane fusion studies. , 1993, Methods in enzymology.

[31]  C. Heimsch DEVELOPMENT OF VASCULAR TISSUES IN BARLEY ROOTS , 1951 .

[32]  F. Bentrup Potassium ion channels in the plasmalemma. , 1990, Physiologia plantarum.

[33]  M. G. Pitman Transport across plant roots , 1982, Quarterly Reviews of Biophysics.

[34]  J. Schroeder K+ transport properties of K+ channels in the plasma membrane of Vicia faba guard cells , 1988, The Journal of general physiology.

[35]  M. Pitman Uptake and Transport of Ions in Barley Seedlings I. Estimation of Chloride Fluxes in Cells of Excised Roots , 1971 .

[36]  F. Maathuis,et al.  Patch clamp studies on root cell vacuoles of a salt-tolerant and a salt-sensitive plantago species : regulation of channel activity by salt stress. , 1990, Plant physiology.

[37]  D. Schachtman,et al.  The k/na selectivity of a cation channel in the plasma membrane of root cells does not differ in salt-tolerant and salt-sensitive wheat species. , 1991, Plant physiology.