| Project/Area Number |
13460151
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | Nara Institute of Science and Technology |
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Principal Investigator |
CHE Fang-sik Nara Institute of Science and Technology, Graduate school of Biological Sciences, Assistant Professor, バイオサイエンス研究科, 助手 (00263442)
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| Co-Investigator(Kenkyū-buntansha) |
IWANO Megumi Nara Institute of Science and Technology, Graduate school of Biological Sciences, Assistant Researcher, バイオサイエンス研究科, 教務職員 (50160130)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2002: ¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 2001: ¥9,800,000 (Direct Cost: ¥9,800,000)
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| Keywords | Hypersensitive cell death / Active oxygen / Rice / Programmed cell death / Flagellin / Electron microscope / Apoptosis / NADPH oxidase / 形質転換体 / 抵抗性反応 / 誘導型発現ベクター |
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| Research Abstract |
To clarify the induction mechanism of the hypersensitive cell death in rice, two researches were performed.
1. Function of rice OsAMPK-_γ on induction of the hypersensitive cell death
Fluorescence differential display was performed using the hypersensitive cell death inducing and non-inducing cultured rice cells. Rice OsAMPK-_γ gene was identified as down regulated genes during the hypersensitive cell death. Anti-sense OsAMRK-_γ gene controlled by DEX-inducible promoter was transformed into cultured rice cells. The hypersensitive cell death was induced in the transgenic cultured cells when the transgenic cultured cells were treated with DEX. The data indicate that downregulation of OsAMRK-_γ gene expression was involved in induction of the hypersensitive cell death in rice.
2. Function of active oxygen species on induction of the hypersensitive cell death
To clarify function of active oxygen species on induction of the hypersensitive cell death, cultured rice cells were inoculated with inc
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ompatible strain of Acidovorax avenae (N1141) and H_2O_2 generation and the hypersensitive cell death were measured. When cultured rice cells were inoculated with the incompatible N1141 strain, H_2O_2 generation was detected before the cell death induction. To examine the location of the H_2O_2 generation, the cultured rice cells inoculated with N1141 strain were treated with Ce (0H) and accumulated Ce was observed by TEM. Plasma membrane was partially convoluted at contact site to bacteria and electron dense material existed between plasma membrane and cell wall. The H_2O_2 generation was dependent on NADPH and inhibited by DPI treatment. Therefore, Osrboh A, NADPH oxidase gene of rice, was cloned from cultured rice cells and anti-sense Osrboh A was transformed into cultured rice cells. When the incompatible strain inoculated to the anti-sense transgenic cells, no H_2O_2 generation was detected and the hypersensitive cell death was also abolished in the transgenic cells, suggesting that H_2O_2 generated by Osrboh A induced the hypersensitive cell death in rice. Less
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