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Related Experiment Videos

Decrease of pH Gradients in Tonoplast Vesicles by NO(3) and Cl: Evidence for H-Coupled Anion Transport.

K S Schumaker1, H Sze

  • 1Botany Department, University of Maryland, College Park, Maryland 20742.

Plant Physiology
|March 1, 1987
PubMed
Summary

Chloride and nitrate ions reduce the pH gradient in plant cell membranes, indicating a novel H(+)-coupled anion transport system. This mechanism may regulate anion storage and mobilization within plant cells.

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Area of Science:

  • Plant cell biology
  • Membrane transport
  • Biochemistry

Background:

  • The tonoplast, or vacuolar membrane, plays a crucial role in maintaining cellular pH homeostasis and storing ions.
  • Understanding anion transport across the tonoplast is essential for comprehending nutrient uptake, storage, and cellular signaling.

Purpose of the Study:

  • To investigate the effect of chloride (Cl-) and nitrate (NO3-) on the pH gradient across the tonoplast.
  • To elucidate the mechanisms of anion-coupled proton transport in plant membranes.

Main Methods:

  • Utilized tonoplast-enriched vesicles to measure pH gradients using [(14)C]methylamine accumulation.
  • Artificially imposed pH gradients via pH jump or K(+) gradient with nigericin to decouple effects from H(+)-ATPase.
  • Investigated the role of 4,4'-Diisothiocyano-2,2'-stilbene disulfonic acid (DIDS) in modulating anion-induced pH changes.

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Main Results:

  • Chloride and nitrate significantly decreased the pH gradient across tonoplast vesicles in an anion concentration-dependent manner.
  • These effects were independent of the tonoplast's H(+)-ATPase, suggesting alternative transport pathways.
  • 4,4'-Diisothiocyano-2,2'-stilbene disulfonic acid partially inhibited the Cl(-)-induced decrease in pH gradient.
  • Similar anion-dependent pH gradient changes were observed in plasma membrane and Golgi fractions.

Conclusions:

  • Identified potential H(+)-coupled anion symport or antiport systems on the tonoplast, plasma membrane, and Golgi.
  • These systems likely facilitate either the mobilization of stored anions (Cl-, NO3-) to the cytoplasm or the transport of anions into the vacuole.
  • The findings suggest a conserved mechanism for H(+)-coupled anion transport across various plant cellular membranes.