| Project/Area Number |
09660063
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Plant nutrition/Soil science
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
MATOH Toru Kyoto University Graduate School of Agriculture Associate Professor, 農学研究科, 助教授 (50157393)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Boron / Cell death / Boron deficiency / tobacco cells / 遺伝子発現 / シロイナズナ |
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| Research Abstract |
Higher plants require boron (B) as an essential nutritional element. How does B work in higher plant cells? To answer this question, the morphological changes of the tobacco BY-2 cultured cells after withdrawal of B from the culture media were followed.
The cells were stained with Fluorescein diacetate (FDA) and Propidium iodide (P1) to discern dead and alive cells. When the tobacco cells were transferred to B-free culture solutions, the cells started to die after 9 hours and at 36 hours more than 80% of the cell died. In dead cells, protoplasm was separated from cell walls as if the cells were plasmolysed and P1 stained evenly the cells. On the other hand, when the cells were killed with 30 mM H202 or 0.2% Triton X-100, the nuclei were stained intensely with P1 while the other part of the protoplasm was not stained. Removal of Ca ions from the culture solution also killed the cells in a similar manner as B did.
To visualize the DNA transcription, the differential display method was applied to B-deficient cells. At 3 and 6 hours after the B removal, any significant and reproducible changes were not detected under the current experimental condition. At 1 8 hours after the treatment, several changes in the DNA transcription were detected. Cloning of the expressed DNA is now under progress.
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