Advertisement

Planta

, Volume 218, Issue 4, pp 615–622 | Cite as

Cellular and whole-plant chloride dynamics in barley: insights into chloride–nitrogen interactions and salinity responses

  • Dev T. Britto
  • Thomas J. Ruth
  • Suzanne Lapi
  • Herbert J. KronzuckerEmail author
Original Article

Abstract

The first analysis of chloride fluxes and compartmentation in a non-excised plant system is presented, examining ten ecologically pertinent conditions. The short-lived radiotracer couple 38Cl/39Cl was used as a Cl tracer in intact barley (Hordeum vulgare L. cv. Klondike) seedlings, which were cultured and investigated under four external [Cl], from abundant (0.1 mM) to potentially toxic (100 mM). Chloride–nitrogen interactions were investigated by varying N source (NO3 or NH4 +) and strength (0.1 or 10 mM), in order to examine, at the subcellular compartmentation level, the antagonism, previously documented at the influx level, between Cl and NO3 , and the potential role of Cl as a counterion for NH4 + under conditions in which cytosolic [NH4 +] is excessive. Cytosolic [Cl] increased with external [Cl] from 6 mM to 360 mM. Cl influx, fluxes to vacuole and shoot, and, in particular, efflux to the external medium, also increased along this gradient. Efflux reached 90% of influx at the highest external [Cl]. Half-times of cytosolic Cl exchange decreased between high-affinity and low-affinity influx conditions. The relationship between cytosolic [Cl] and shoot flux indicated the presence of a saturable low-affinity transport system (‘SLATS’) responsible for xylem loading of Cl. N source strongly influenced Cl flux to the vacuole, and moderately influenced Cl influx and shoot flux, whereas efflux and half-time were insensitive to N source. Cytosolic pool sizes were not strongly or consistently influenced by N source, indicating the low potential for Cl to act as a counterion to hyperaccumulating NH4 +. We discuss our results in relation to salinity responses in cereals.

Keywords

Compartmental analysis Efflux Hordeum Ion transport Salinity Translocation 

Abbreviations

[Cl]cyt

cytosolic chloride concentration

[Cl]o

external chloride concentration

Notes

Acknowledgments

We thank A.D.M. Glass and J. Dainty for helpful discussion of the manuscript, A.D.M. Glass for the generous provision of laboratory space and equipment, and the Natural Sciences and Engineering Resource Council of Canada (NSERC) for financial support.

References

  1. Adler PR, Wilcox GE (1995) Ammonium increases the net rate of sodium influx and partitioning to the leaf of muskmelon. J Plant Nutr 18:1951–1962Google Scholar
  2. Apse MP, Blumwald E (2002) Engineering salt tolerance in plants. Curr Opin Biotechnol 13:146–150CrossRefPubMedGoogle Scholar
  3. Bar Y, Apelbaum A, Kafkafi U, Goren R (1997) Relationship between chloride and nitrate and its effect on growth and mineral composition of avocado and citrus plants. J Plant Nutr 20:715–731Google Scholar
  4. Binzel ML, Hess FD, Bressan RA, Hasegawa PM (1988) Intracellular compartmentation of ions in salt adapted tobacco cells. Plant Physiol 86:607–614Google Scholar
  5. Britto DT, Kronzucker HJ (2001) Constancy of nitrogen turnover kinetics in the plant cell: insights into the integration of subcellular N fluxes. Planta 213:175–181CrossRefPubMedGoogle Scholar
  6. Britto DT, Kronzucker HJ (2003) Ion fluxes and cytosolic pool sizes: examining fundamental relationships in transmembrane flux regulation. Planta 217:490–497PubMedGoogle Scholar
  7. Britto DT, Siddiqi MY, Glass ADM, Kronzucker HJ (2001) Futile transmembrane NH4 + cycling: a cellular hypothesis to explain ammonium toxicity in plants. Proc Natl Acad Sci USA 98:4255–4258CrossRefPubMedGoogle Scholar
  8. Collins JC, Abbas MA (1985) Ion and water transport in seedlings of mustard (Sinapis alba L). New Phytol 99:195–202Google Scholar
  9. Cram WJ (1968) Compartmentation and exchange of chloride in carrot root tissue. Biochim Biophys Acta 163:339–353CrossRefPubMedGoogle Scholar
  10. Cram WJ (1973) Chloride fluxes in cells of isolated root cortex of Zea mays. Aust J Biol Sci 26:757–779Google Scholar
  11. Cram WJ (1983) Chloride accumulation as a homeostatic system—set points and perturbations—the physiological significance of influx isotherms, temperature effects and the influence of plant-growth substances. J Exp Bot 34:1484–1502Google Scholar
  12. Cram WJ, Laties GG (1971) Use of short-term and quasi-steady influx in estimating plasmalemma and tonoplast influx in barley root cells at various external and internal chloride concentrations. Aust J Biol Sci 24:633–646Google Scholar
  13. Deane-Drummond CE (1986) A comparison of regulatory effects of chloride on nitrate uptake, and of nitrate on chloride uptake into Pisum sativum seedlings. Physiol Plant 66:115–121Google Scholar
  14. Downton WJS, Millhouse J (1983) Turgor maintenance during salt stress prevents loss of variable fluorescence in grapevine leaves. Plant Sci Lett 31:1–7CrossRefGoogle Scholar
  15. Drew MC, Saker LR (1984) Uptake and long-distance transport of phosphate, potassium and chloride in relation to internal ion concentrations in barley—evidence of non-allosteric regulation. Planta 160:500–507Google Scholar
  16. Eaton FM (1966) Chlorine. In: Chapman HD (ed) Diagnostic criteria for plants and soils. University of California Division of Agricultural Sciences, Riverside, pp 98–135Google Scholar
  17. Elzam OE, Epstein E, Rains DW (1964) Ion transport kinetics in plant tissue—complexity of chloride absorption isotherm. Biochem Biophys Res Commun 15:273–276PubMedGoogle Scholar
  18. Felle HH (1994) The H+/Cl symporter in root-hair cells of Sinapis alba. Plant Physiol 106:1131–1136PubMedGoogle Scholar
  19. Flowers TJ (1988) Chloride as a nutrient and as an osmoticum. Adv Plant Nutr 3:55–78Google Scholar
  20. Flowers TJ, Hajibagheri MA (2001) Salinity tolerance in Hordeum vulgare: ion concentrations in root cells of cultivars differing in salt tolerance. Plant Soil 231:1–9CrossRefGoogle Scholar
  21. Flowers TJ, Troke PF, Yeo AR (1977) Mechanism of salt tolerance in halophytes. Annu Rev Plant Physiol Plant Mol Biol 28:89–121CrossRefGoogle Scholar
  22. Gerson DF, Poole RJ (1972) Chloride accumulation by mung bean root tips—low affinity active transport-system at plasmalemma. Plant Physiol 50:603–607Google Scholar
  23. Gibson TS, Speirs J, Brady CJ (1984) Salt tolerance in plants. 2. In vitro translation of mRNAs from salt-tolerant and salt-sensitive plants on wheat-germ ribosomes—responses to ions and compatible organic solutes. Plant Cell Environ 7:579–587Google Scholar
  24. Gimmler H, Kaaden R, Kirchner U, Weyand A (1984) The chloride sensitivity of Dunaliella parva enzymes. Z Pflanzenphysiol 114:131–150Google Scholar
  25. Glass ADM, Siddiqi MY (1985) Nitrate inhibition of chloride influx in barley—implications for a proposed chloride homeostat. J Exp Bot 36:556–566Google Scholar
  26. Greenway H (1965) Plant responses to saline substrates. 4. Chloride uptake by Hordeum vulgare as affected by inhibitors transpiration and nutrients in medium. Aust J Biol Sci 18:249–268Google Scholar
  27. Hajibagheri MA, Flowers TJ, Collins JC, Yeo AR (1988) A comparison of the methods of X-ray microanalysis, compartmental analysis and longitudinal ion profiles to estimate cytoplasmic ion concentrations in 2 maize varieties. J Exp Bot 39:279–290Google Scholar
  28. Hajibagheri MA, Yeo AR, Flowers TJ, Collins JC (1989) Salinity resistance in Zea mays—fluxes of potassium, sodium and chloride, cytoplasmic concentrations and microsomal membrane lipids. Plant Cell Environ 12:753–757Google Scholar
  29. Harvey DMR (1985) The effects of salinity on ion concentrations within the root-cells of Zea mays L. Planta 165:242–248Google Scholar
  30. Harvey DMR, Thorpe JR (1986) Some observations on the effects of salinity on ion distributions and cell ultrastructure in wheat leaf mesophyll cells. J Exp Bot 37:1–7Google Scholar
  31. Hill AE (1970) Ion and water transport in Limonium. 3. Time constants of transport system. Biochim Biophys Acta 196:66–72CrossRefPubMedGoogle Scholar
  32. Huang CX, van Steveninck RFM (1989) Longitudinal and transverse profiles of K+ and Cl concentration in low-salt and high-salt barley roots. New Phytol 112:475–480Google Scholar
  33. Jacoby B, Rudich B (1980) Proton–chloride symport in barley roots. Ann Bot 46:493–498Google Scholar
  34. Jefferies RL, Jensen A, Bazely D (1983) The biology of the annual Salicornia Europaea agg at the limits of its range in Hudson Bay. Can J Bot 61:762–773Google Scholar
  35. Jeschke WD, Klagges S, Hilpert A, Bhatti AS, Sarwar G (1995) Partitioning and flows of ions and nutrients in salt-treated plants of Leptochloa fusca L. Kunth.1. Cations and chloride. New Phytol 130:23–35Google Scholar
  36. Kafkafi U, Valoras N, Letey J (1982) Chloride interaction with nitrate and phosphate nutrition in tomato (Lycopersicon esculentum L). J Plant Nutr 5:1369–1385Google Scholar
  37. Kronzucker HJ, Siddiqi MY, Glass ADM (1995) Analysis of 13NH4 + efflux in spruce roots—a test case for phase identification in compartmental analysis. Plant Physiol 109:481–490PubMedGoogle Scholar
  38. Kronzucker HJ, Britto DT, Davenport RJ, Tester M (2001) Ammonium toxicity and the real cost of transport. Trends Plant Sci 6:335–337CrossRefPubMedGoogle Scholar
  39. Kronzucker HJ, Szczerba MW, Britto DT (2003) Cytosolic potassium homeostasis revisited: 42K-tracer analysis in Hordeum vulgare L. reveals set-point variations in [K+]. Planta 217:540–546PubMedGoogle Scholar
  40. Lee RB, Clarkson DT (1986) 13N studies of nitrate fluxes in barley roots. 1. Compartmental analysis from measurements of 13N efflux. J Exp Bot 37:1753–1767Google Scholar
  41. Macklon AES (1975) Cortical cell fluxes and transport to stele in excised root segments of Allium cepa L.1. Potassium, sodium and chloride. Planta 122:109–130Google Scholar
  42. Macklon AES (1976) An examination, by compartmental flux analysis, of the development of sodium and chloride absorption capacities in beetroot disks. J Exp Bot 27:651–657Google Scholar
  43. Memon AR, Saccomani M, Glass ADM (1985) Efficiency of potassium utilization by barley varieties—the role of subcellular compartmentation. J Exp Bot 36:1860–1876Google Scholar
  44. Noble CL, Rogers ME (1992) Arguments for the use of physiological criteria for improving the salt tolerance in crops. Plant Soil 146:99–107Google Scholar
  45. Oertli JJ (1968) Extracellular salt accumulation a possible mechanism of salt injury in plants. Agrochimica 12:461–469Google Scholar
  46. Peuke AD, Jeschke WD, Hartung W (1998) Foliar application of nitrate or ammonium as sole nitrogen supply in Ricinus communis—II. The flows of cations, chloride and abscisic acid. New Phytol 140:625–636CrossRefGoogle Scholar
  47. Pierce WS, Higinbotham N (1970) Compartments and fluxes of K+, Na+ and Cl in Avena coleoptile cells. Plant Physiol 46:666–673Google Scholar
  48. Pitman MG (1971) Uptake and transport of ions in barley seedlings.1. Estimation of chloride fluxes in cells of excised roots. Aust J Biol Sci 24:407–421Google Scholar
  49. Sanders D (1984) Gradient-coupled chloride transport in plant cells. In: Gerenscser GA (ed) Chloride transport coupling in biological membranes and epithelia. Elsevier, Amsterdam, pp 63–120Google Scholar
  50. Seemann JR, Critchley C (1985) Effects of salt stress on the growth, ion content, stomatal behavior and photosynthetic capacity of a salt-sensitive species, Phaseolus vulgaris L. Planta 164:151–162Google Scholar
  51. Siddiqi MY, Glass ADM, Ruth TJ (1991) Studies of the uptake of nitrate in barley. 3. Compartmentation of NO3 . J Exp Bot 42:1455–1463Google Scholar
  52. Speer M, Brune A, Kaiser WM (1994) Replacement of nitrate by ammonium as the nitrogen source increases the salt sensitivity of pea plants.1. Ion concentrations in roots and leaves. Plant Cell Environ 17:1215–1221Google Scholar
  53. Wang MY, Siddiqi MY, Ruth TJ, Glass ADM (1993) Ammonium uptake by rice roots 1. Fluxes and subcellular distribution of 13NH4 +. Plant Physiol 103:1249–1258CrossRefPubMedGoogle Scholar
  54. White PJ, Broadley MR (2001) Chloride in soils and its uptake and movement within the plant: a review. Ann Bot 88:967–988CrossRefGoogle Scholar
  55. Xu GH, Magen H, Tarchitzky J, Kafkafi U (2000) Advances in chloride nutrition of plants. Adv Agron 68:97–150Google Scholar
  56. Zhong WJ, Kaiser W, Kohler J, Bauer-Ruckdeschel H, Komor E (1998) Phloem loading of inorganic cations and anions by the seedling of Ricinus communis L. J Plant Physiol 152:328–335Google Scholar
  57. Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Dev T. Britto
    • 1
  • Thomas J. Ruth
    • 2
  • Suzanne Lapi
    • 2
    • 3
  • Herbert J. Kronzucker
    • 1
    Email author
  1. 1.Department of Life Sciences, Department of BotanyUniversity of TorontoTorontoCanada
  2. 2.Tri-University Meson Facility (TRIUMF)VancouverCanada
  3. 3.Simon Fraser UniversityBurnabyCanada

Personalised recommendations