Effects of anion channel blockers on xylem nitrate transport in barley seedlings

Abstract The effects of anion channel blockers, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) and anthracene-9-carboxylic acid (A-9-C), on the uptake and xylem transport of nitrate were investigated in intact barley (Hordeum vulgare L.) seedlings using 13NO3 − and 15NO3 − as tracers. The seedling roots were pretreated with either blocker for 1 h and then 13NO3 − or 15NO3 − was supplied to the medium. Real-time images of 13N accumulation in shoots were monitored during the first 30 min using Positron Emitting Tracer Imaging Systems (PETIS). The radioactivity in the shoots of the plants treated with either blocker was about half of that in the shoots of the control plants. Analysis of the distribution of 13N in whole seedlings using a Bioimaging Analyzer System (BAS) showed a 13N-accumulation in both shoots and roots of the control plants but reduced 13N-levels in the shoots of the plants treated with either blocker. Nitrate concentrations in the xylem sap were significantly reduced by the application of either blocker while the net uptake of 15NO3 − was not or slightly influenced by the treatment with blockers. The translocation and uptake of chloride were also significantly reduced by the treatment with A-9-C but not with DIDS. These results suggested that anion channels contributed to xylem loading of nitrate in barley plants.

[1]  H. Hayashi,et al.  Detection and characterization of nitrogen circulation through the sieve tubes and xylem vessels of rice plants , 1997, Plant and Soil.

[2]  J. Schroeder Anion channels as central mechanisms for signal transduction in guard cells and putative functions in roots for plant-soil interactions , 1995, Plant Molecular Biology.

[3]  Susan J. Smith,et al.  The mechanism of nitrate transport across the tonoplast of barley root cells , 1992, Planta.

[4]  A. D. Tomos,et al.  Compartmental nitrate concentrations in barley root cells measured with nitrate-selective microelectrodes and by single-cell sap sampling , 1991, Planta.

[5]  S. Tyerman,et al.  A channel that allows inwardly directed fluxes of anions in protoplasts derived from wheat roots , 2004, Planta.

[6]  R. L. Warner,et al.  Role of xylem sap nitrate in the regulation of nitrate reductase gene expression in leaves of barley (Hordeum vulgare L.) seedlings , 2002 .

[7]  Hiroshi Uchida,et al.  Real Time Visualization of 13N-Translocation in Rice under Different Environmental Conditions Using Positron Emitting Tracer Imaging System , 2001 .

[8]  N. Ohtake,et al.  Rapid N transport to pods and seeds in N-deficient soybean plants. , 2001, Journal of experimental botany.

[9]  C. Maurel,et al.  Anion channels in higher plants: functional characterization, molecular structure and physiological role. , 2000, Biochimica et biophysica acta.

[10]  J. Frachisse,et al.  Characterization of a nitrate-permeable channel able to mediate sustained anion efflux in hypocotyl cells from Arabidopsis thaliana. , 2000, The Plant journal : for cell and molecular biology.

[11]  J. Boyer,et al.  Passive nitrate transport by root plasma membrane vesicles exhibits an acidic optimal pH like the H(+)-ATPase. , 2000, Plant physiology.

[12]  K. Raschke,et al.  The delivery of salts to the xylem. Three types of anion conductance in the plasmalemma of the xylem parenchyma of roots of barley. , 2000, Plant physiology.

[13]  S. Thomine,et al.  Sulfate is both a substrate and an activator of the voltage-dependent anion channel of Arabidopsis hypocotyl cells. , 1999, Plant physiology.

[14]  孝 佐藤,et al.  根粒非着生と根粒着生ダイズにおける13NO3-と15NO3-を用いた硝酸吸収と移行の解析 , 1999 .

[15]  D. Clarkson,et al.  Nitrate and ammonium nutrition of plants: physiological and molecular perspectives , 1999 .

[16]  N. Ohtake,et al.  Determination of leghemoglobin components and xylem sap composition by capillary electrophoresis in hypernodulation soybean mutants cultivated in the field , 1998 .

[17]  N. Crawford,et al.  Molecular and physiological aspects of nitrate uptake in plants , 1998 .

[18]  S. Thomine,et al.  Anion-Channel Blockers Interfere with Auxin Responses in Dark-Grown Arabidopsis Hypocotyls , 1997, Plant physiology.

[19]  Hiroshi Uchida,et al.  Uptake and transport of positron-emitting tracer (18F) in plants☆ , 1997 .

[20]  P. Pouliquin,et al.  In vitro study of passive nitrate transport by native and reconstituted plasma membrane vesicles from corn root cells. , 1997, Biochimica et biophysica acta.

[21]  W. Vaalburg,et al.  Nitrate and ammonium influxes in soybean (Glycine max) roots: direct comparison of 13N and 15N tracing , 1996 .

[22]  J. Schroeder,et al.  Anion-Channel Blockers Inhibit S-Type Anion Channels and Abscisic Acid Responses in Guard Cells , 1995, Plant physiology.

[23]  R. L. Warner,et al.  Expression of NADH-Specific and NAD(P)H-Bispecific Nitrate Reductase Genes in Response to Nitrate in Barley , 1995, Plant physiology.

[24]  J. Schroeder,et al.  Anion Selectivity of Slow Anion Channels in the Plasma Membrane of Guard Cells (Large Nitrate Permeability) , 1994, Plant physiology.

[25]  B. Touraine,et al.  Effects of NaCl on Flows of N and Mineral Ions and on NO3- Reduction Rate within Whole Plants of Salt-Sensitive Bean and Salt-Tolerant Cotton , 1994, Plant physiology.

[26]  L. Wegner,et al.  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 , 1994, Plant physiology.

[27]  J. Schroeder,et al.  Identification of High-Affinity Slow Anion Channel Blockers and Evidence for Stomatal Regulation by Slow Anion Channels in Guard Cells. , 1993, The Plant cell.

[28]  David T. Clarkson,et al.  Roots and the Delivery of Solutes to the Xylem , 1993 .

[29]  C. Schobert,et al.  Transport of Nitrate and Ammonium into the Phloem and the Xylem of Ricinus communis seedlings , 1992 .

[30]  R. L. Warner,et al.  Synthesis and degradation of barley nitrate reductase. , 1983, Plant physiology.

[31]  T. Ohyama,et al.  Assimilation and transport of nitrogenous compounds originated from 15N2 fixation and 15NO2 absorption , 1979 .

[32]  J. Boyer,et al.  Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: I. Regulation by Nitrate Flux. , 1976, Plant physiology.