Characterization of the TaALMT1 protein as an Al3+-activated anion channel in transformed tobacco (Nicotiana Tabacum L.) cells

Wen Hao Zhang, Peter R. Ryan, Takayuki Sasaki, Yoko Yamamoto, Wendy Sullivan, Steve D. Tyerman

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

TaALMT1 encodes a putative transport protein associated with Al 3+-activated efflux of malate from wheat root apices. We expressed TaALMT1 in Nicotiana tabacum L. suspension cells and conducted a detailed functional analysis. Protoplasts were isolated for patch-clamping from cells expressing TaALMT1 and from control cells (empty vector transformed). With malate2- as the permeant anion in the protoplast, an inward current (anion efflux) that reversed at positive potentials was observed in protoplasts expressing TaALMT1 in the absence of Al3+. This current was sensitive to the anion channel antagonist niflumate, but insensitive to Gd3+. External AlCl3 (50 μM), but not La3+ and Gd 3+, increased the inward current in TaALMT1-transformed protoplasts. The inward current was highly selective to malate over nitrate and chloride (Pmal >> PNO3 < PCl, P mal/PCl <18, +/-Al3+), under conditions with higher anion concentration internally than externally. The anion currents displayed a voltage and time dependent deactivation at negative voltages. Voltage ramps revealed that inward rectification was caused by the imposed anion gradients. Single channels with conductances between 10 and 17 pS were associated with the deactivation of the current at negative voltages, agreeing with estimates from voltage ramps. This study of the electrophysiological function of the TaALMT1 protein in a plant heterologous expression system provides the first direct evidence that TaALMT1 functions as an Al 3+-activated malate2- channel. We show that the Al 3+-activated currents measured in TaALMT1-transformed tobacco cells are identical to the Al3+-activated currents observed in the root cells of wheat, indicating that TaALMT1 alone is likely to be responsible for those endogenous currents.

Original languageEnglish
Pages (from-to)1316-1330
Number of pages15
JournalPlant and Cell Physiology
Volume49
Issue number9
DOIs
Publication statusPublished - Sep 1 2008

Keywords

  • ALMT1
  • Al-gated
  • Aluminium
  • Anion channel
  • Malate
  • Patch-clamp

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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