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Plant and Cell Physiology, 2000, Vol. 41, No. 7 840-849
© 2000 Oxford University Press

Mechanism of the Chilling-Induced Decrease in Proton Pumping across the Tonoplast of Rice Cells

Kunihiro Kasamo1,3, Mineo Yamaguchi1 and Yoshiyuki Nakamura2

1 Research Institute for Bioresources, Okayama University, Kurashiki, Okayama, 710-0046 Japan 2 Chugoku National Agricultural Experiment Station, Fukuyama, Hiroshima, 721-8514 Japan

The ATP-generated proton pumping across tonoplast vesicles from chilling-sensitive Boro rice (Oryza sativa L. var. Boro) cultured cells was markedly decreased by chilling at 5°C for 3 d. The membrane fluidity of core hydrophobic and surface hydrophilic regions of the lipid bilayer was measured by steady-state fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene and trimethylammonium 1,6-diphenyl-1,3,5-hexatriene and by electron spin resonance spectroscopy of 16- and 5-doxyl stearic acid, respectively. The fluidity of the surface region of the lipid bilayer of the tonoplast vesicles decreased by chilling. The fluidity of the surface region of the liposomes and the proton pumping across the reconstituted proteoliposomes with tonoplast H+-ATPase decreased with increasing content of the glycolipids. The proton pumping across chimera proteoliposomes was reduced by chilling only when it was reconstituted in the presence of tonoplast glycolipids from chilled Boro cells. These data suggest that the reduction in ATP-generated proton pumping across the tonoplast by chilling is due to the decrease in the fluidity of the surface region of the lipid bilayer of the tonoplast, which is caused by the changes in glycolipids.

3 Corresponding author: E-mail, kasamo@rib.okayama-u.ac.jp; Fax, +81-(0)86-434-1221.


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