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Plant and Soil

, Volume 142, Issue 2, pp 203–210 | Cite as

Soil humic substances affect transport properties of tonoplast vesicles isolated from oat roots

  • R. Pinton
  • Z. Varanini
  • G. Vizzotto
  • A. Maggioni
Article

Abstract

The effect of a low molecular size (<5 KDa) humic fraction, essentially fulvic acids, on microsomal and tonoplast ion-stimulated ATPase activity was studied. After 20 min of pre-incubation with microsomal vesicles from oat roots, humic substances at organic C concentration of up to 0.5 μg cm-3 increased KCl-stimulated ATPase activity, while they inhibited enzyme activity at higher concentrations. Cl--stimulated ATPase activity of tightly sealed tonoplast-enriched vesicles was similarly affected by <5 KDa humic substances. This behaviour was not observed when gramicidin D was added to the assay medium. Proton transport by vesicles incubated up to 5 min with <5 KDa humic molecules was affected in a concentration-dependent manner, strongly resembling that observed for ATP hydrolysis, whereas it was severely reduced when the assay conditions were close to those used for measuring ATP hydrolysis (20 min pre-incubation of vesicles with humic substances). The transmembrane electrical potential was negatively affected, irrespective of the concentration of humic molecules. Furthermore, a 15-min pre-incubation strongly reduced the formation of a potential gradient. The size and concentrations of humic substances employed make an interaction with the vacuolar membrane of root cells plausible. The results show that the main target of humic molecules is the electrical membrane potential and suggest a possible way of interference of these naturally occurring substances with the biochemical mechanisms involved in plant mineral nutrition.

Key words

acridine orange Avena sativa L. humic substances oxonol VI proton gradient tonoplast ATPase transmembrane potential 

Abbreviations

BSA

bovine serum albumin

BTP

bis-tris-propane (1,3-bis(tris(hydroxy-methyl)-methylamino)-propane)

DTT

dithiothreitol

EGTA

ethylene glycol bis(β-aminoethyl ether)-N,N′-tetracetic acid

Hepes

(N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid))

IDA

iminodiacetic acid

oxonol VI

bis (3-propyl-5-oxoisoxazol-4-yl)pentamethine oxonol

Tris

(tris-(hydroxy-methyl)aminomethane)

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Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • R. Pinton
    • 1
  • Z. Varanini
    • 1
  • G. Vizzotto
    • 1
  • A. Maggioni
    • 1
  1. 1.Dipartimento di Produzione Vegetale e Tecnologie AgrarieUniversity of UdineUdineItaly

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