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Plant and Cell Physiology, 1994, Vol. 35, No. 5 829-836
© 1994

Effects of Gamma-Irradiation on the Activity of Tonoplast H+-ATPase from Potato Tubers (Solanum tuberosum L.)

Setsuko Todoriki, Toru Hayashi, Yoshiyuki Nakamura and Kunihiro Kasamo

National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries 2-1-2 Kannondai, Tsukuba, Ibaraki, 305 Japan

Tonoplast vesicles were prepared from potato tubers (Solarium tuberosum L.) on a step gradient (0% and 6%, w/w) of dextran T-70 to clarify the mechanism by which the tonoplast H+-ATPase is inactivated by gamma-irradiation. H+-ATPase activity and H+ -pumping were examined after irradiation of tubers (in vivo irradiation) and of isolated tonoplast vesicles (in vitro irradiation) at doses up to 1.0 kGy. Both in vivo irradiation and in vitro irradiation resulted in significant decreases in ATPase and H+-pumping activities. The ATPase and H+-pumping activities 12 h after irradiation were much lower than those 2 h after irradiation. Solubilized H+-ATPase was inactivated, in a dose-dependent manner, by irradiation (enzyme irradiation) to a greater extent than was observed after in vitro irradiation or in vivo irradiation. The activity of ATPase 12 h after enzyme irradiation was almost the same as it was 2 h after enzyme irradiation. The free fatty acid content of vacuolar membranes was increased by in vivo irradiation and by in vitro irradiation with an accompanying decrease in tonoplast H+-ATPase activity. Lipids from irradiated tonoplasts had a considerable inhibitory effect on the activity of solubilized H+-ATPase. This result suggests that the direct inactivation of H+-ATPase in potato tonoplast by gamma-irradiation is augmented by the effects of deterioration of membrane lipids that is induced by the irradiation.

(Received December 21, 1994; Accepted May 16, 1994)
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