The elucidation of the low-temperature stress mechanism using reconstituted vesicles with tonoplast H^+-ATPase

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Proton pumping across the tonoplast was markedly suppressed by chilling. In order to clarify the mechanism of chilling-induced depression of proton-pumping across tonoplast. we reconstituted chimera proteoliposomes with H^+-ATPase solubilized from the tonoplast of chilling insensitive rice (var. Nipponbare) without chilling and individual or combined lipids extracted from the tonoplast of chilling-sensitive rice (var. Boro) cells with or without chilling at 5 ℃. We further examined the relation between membrane fluidity and chilling.As the result, the chilling-induced decrease in proton pumping across the tonoplast is due to the decrease in the fluidity of the hydrophilic surface region of the lipid bilayer of the tonoplast, which is caused by glycolipids, such as glucocerebroside and acylsteryl glucoside.