| Project/Area Number |
63540538
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物生理学
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| Research Institution | University of Shizuoka |
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Principal Investigator |
MIYOSHI Yasuhiro University of Shizuoka, Dept. of Biology, Professor, 国際関係学部, 教授 (40008165)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Phytochrome / Protoplast / Red / Far-Red Irradiation / Fura 2 / Electroporation / ATPase Activity / Intracellular Ca^<2+> CONCENTRATION / カルシウムポンプ / プロトンポンプ / 光可逆的 / 水性二層分配法 / シールドベシクル / 細胞内Caイオン濃度 |
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| Research Abstract |
The temporal changes of intracellular Ca^<2+> concentration were measured following red/far-red light irradiation, using protoplasts of Vigna mungo and Pisum sativum isolated from 5-day-old dark-grown seedlings. The intracellular Ca^<2+> concentration was measured by spectrophotometry using fluorogenic calcium probe Fura2-AM. The reagent was injected into the protoplasts by electroporation in 20 muM Fura2-AM solution followed by 30 min. Of incubation at 25^ C in the same solution. The intracellular Ca^<2+> concentrations were riversibly changed by red/far-red light irradiation. The degrees of increase and decrease of the Ca^<2+> concentration depended on Ca^<2+> and H^+ concentrations in the suspension medium. Therefore, it was suggested that the intracellular Ca^<2+> concentration change by red/far-red irradiation was brought not across the membranes of organella but through plasma membrane.
ATPase activity in microsomal and mitochondrial fractions isolated from 5-day-old dark-grown seedlings of Vigna mungo, Avena sativa and Pisum sativum was measured. The ATPase activity of the mitochondrial fractions per mg protein was the highest in Pisum sativum and the lowest in Vigna mungo. The ATPase activity of the microsomal fractions was very low. The ATPase activity of the mitochondrial fraction of Pisum sativum was changed reversibly by red/far-red irradiation without any phytochrome enrichment process. Red light irradiation decreased the ATPase activity. Far-rd light recovered the activity to the dark level. The phytochrome-dependent ATPase activity change in the mitochondrial fractions depended more strongly on the concentration of H^+ in the medium than that of Ca^<2+>.
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