Induction signaling and its mechanism of hypersensitive cell death which is thought to be the rice defense responses

诱导信号及其过敏细胞死亡机制（被认为是水稻的防御反应）

• 批准号: 13460151
• 负责人: CHE Fang-sik
• 金额: $ 9.98万
• 依托单位: Nara Institute of Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460151/
• 关键词: 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

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 deathFluorescence 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 deathTo clarify function of active oxygen species on induction of the hypersensitive cell death, cultured rice cells were inoculated with inc … More 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

为了阐明水稻超敏性细胞死亡的诱导机制，本研究进行了两个方面的研究.水稻OsAMPK-γ在诱导过敏性细胞死亡中的作用利用诱导和未诱导过敏性细胞死亡的水稻培养细胞进行荧光差异显示。水稻OsAMPK-γ基因被鉴定为在过敏性细胞死亡过程中表达下调的基因。将DEX诱导型启动子控制的反义OsAMRK-γ基因转化水稻细胞。当转基因培养细胞用地塞米松处理时，可诱导转基因培养细胞发生超敏性细胞死亡。结果表明，OsAMRK-_γ基因表达的下调参与了水稻过敏性细胞死亡的诱导.活性氧在诱导水稻细胞过敏性死亡中的作用为了阐明活性氧在诱导水稻细胞过敏性死亡中的作用，本研究以水稻细胞为材料，接种了不同浓度的外源基因，研究了外源基因对水稻细胞过敏性死亡的影响。 ...更多信息 测定了燕麦食酸菌（Acidovorax avenae，N1141）的H_2O_2产生量和过敏性细胞死亡率。用不亲和菌株N1141接种水稻培养细胞，在诱导细胞死亡之前就有H_2O_2产生。用Ce（OH）处理接种N1141菌株的水稻细胞，透射电镜观察Ce的积累，以确定H_2O_2产生的位置。细胞质膜与细菌接触处部分卷曲，细胞壁与质膜之间存在电子致密物质。H_2O_2的产生依赖于NADPH，DPI抑制H_2O_2的产生。因此，从水稻培养细胞中克隆了水稻NADPH氧化酶基因Osrboh A，并将反义Osrboh A转化到水稻培养细胞中。将不亲和菌株接种到反义转基因细胞上，没有检测到H_2O_2的产生，转基因细胞的过敏性死亡也被消除，表明Osrboh A产生的H_2O_2诱导了水稻过敏性细胞死亡。少

项目成果

Fang-Sik Che: "Identification of novel gene which are differentially expressed in compatible and incompatible interactions between rice and Pseudomonas avenae"Plant Science. 162. 449-458 (2002)

Fang-Sik Che：“水稻和燕麦假单胞菌之间相容和不相容相互作用中差异表达的新基因的鉴定”植物科学。

Megumi Iwano: "Electron microscopic analysis of the H_2O_2 accumulation proceeding to hypersensitive cell death in cultured rice cells induced by an incompatible strain of Pseudomonas avanae"Molecular Plant Pathology. 3. 1-8 (2002)

Megumi Iwano：“对由不相容的 avanae 假单胞菌菌株诱导的培养水稻细胞中 H_2O_2 积累导致过敏性细胞死亡的电子显微镜分析”《分子植物病理学》。

Megumi Iwano et al.: "Electron microscopic analysis of the H_2O_2 accumulation preceding to hypersensitive cell death in cultured rice cells induced by an incompatible strain of Pseudomonas avenae"Mol. Plant Pathol.. 3. 1-8 (2002)

Megumi Iwano 等人：“由不相容的燕麦假单胞菌菌株诱导的培养水稻细胞中过敏性细胞死亡之前的 H_2O_2 积累的电子显微镜分析”Mol。

Megumi Iwano. 他: "Electron microscopic analysis of the H_2O_2 accumulation preceding to hypersensitive cell death in cultured rice cells induced by an incompatible strain of Pseudomonas avenae."Molecular Plant Pathology. 3. 1-8 (2003)

Megumi Iwano. 等人：“由不相容的燕麦假单胞菌菌株诱导的培养水稻细胞中过敏性细胞死亡之前的 H_2O_2 积累的电子显微镜分析”。《分子植物病理学》。

Noriko Tanaka: "Flagellin from incompatible strain of Acidovorax avanae mediates H2O2 generation accompanying hypersensitive cell death and PAL, Cht-1, and PBZ1 expression, but not LOX expression in rice"Molecular Plant Microbe-Interactions. (印刷中). (2003)

Noriko Tanaka：“来自 Acidovorax avanae 的不相容菌株的鞭毛蛋白介导 H2O2 的产生，并伴随过敏性细胞死亡和 PAL、Cht-1 和 PBZ1 表达，但不介导水稻中的 LOX 表达”（出版中）。