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The mechanism of nickel uptake into vacuoles isolated from leaf tissue of Alyssum lesbiacum was investigated to help understand the ability of this species to hyperaccumulate Ni. An imaging system was designed to monitor Ni uptake by single vacuoles using the metal-sensitive fluorescent dye, Newport Green. Nickel uptake into isolated vacuoles from leaf tissue of A. lesbiacum was enhanced by the presence of Mg/ATP, presumably via energisation of the vacuolar H(+)-ATPase (V-ATPase). This ATP-stimulated Ni uptake was abolished by bafilomycin (a diagnostic inhibitor of the V-ATPase) and by dissipation of the transmembrane pH difference with an uncoupler. These observations are consistent with Ni(2+)/nH(+) antiport activity at the tonoplast driven by a proton electrochemical gradient established by the V-ATPase, which would provide a mechanism for secondary active transport of Ni(2+) into the vacuole. This study provides insights into the molecular basis of Ni tolerance in Alyssum, and may aid in the identification of genes involved in Ni hyperaccumulation.

Original publication

DOI

10.1111/j.1438-8677.2008.00080.x

Type

Journal article

Journal

Plant Biol (Stuttg)

Publication Date

11/2008

Volume

10

Pages

746 - 753

Keywords

Adenosine Triphosphate, Antiporters, Brassicaceae, Fluorescent Dyes, Hydrogen-Ion Concentration, Macrolides, Microscopy, Fluorescence, Nickel, Vacuolar Proton-Translocating ATPases, Vacuoles