Molecular and physiological mechanism of oxidative burst in relation to signal transduction for defense response of plant to infection

氧化爆发与植物感染防御反应信号转导相关的分子和生理机制

• 批准号: 09460027
• 负责人: DOKE Noriyuki
• 金额: $ 8.96万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460027/
• 关键词: Oxidative burst; O_2 generating NADPH oxidse; Solubilization of NADPH oxidase; Isolation of NADPH oxidase; gp91^<phox> homologus cDNA; Function of oxidative burst; Expression of defense-related genes; Potato plant; 防御関連遺 遺伝子の発現; O_2NADPH酸化酵素; 酸素

Plants cause induced defense response to pathogens or elicitor. Rapid activation of superoxide generating reaction (Oxidative burst : OXB) is induced at the beginning of induction of the defense response. This reaction system was shown to be dependent on an NADPH oxidase in plasma membrane regulated by signal trasnduction system involving Ca^<2+>, calumodulin, protein kinase, etc. In the present research, isolation and characterization of the NADPH oxidase, signal transduction stimulated by OXB and OXB-induced induction of defense-related genes were investigated with the following results. 1. Time course increase in potential for response with OXB on the sliced surface during aging after slicing of potato tuber. 2. Induction of sub-systemic OXB at aged sliced surface distant from elicitor treated side with local OXB.3. Kinetics of O_2 generating NADPH oxidase in isolated plasma membrane fraction of potato tuber. 4. Various inhibitor for Ca^<2+> signaling or protein kinase inhibited ind … More uctions of local and sub-systemic. 5. Solubirization of active 0_2 generating NADPH oxidase from plasma membrane by detergents. 6. Detection of several active bands in polyacril amid gel after electrophoresis of solubilized NADPH oxidase. 7. Cloning of cDNA homologous with gp91p^<phox> of human leukocytes and detection of homology with those from rice and Arabidopsis plants. 8. Detection of signal by antibody against p67^<phax> from human leukocytes in the fraction on SDS-PAGE of soluble protein from potato tuber tissues. 9. Inhibition by Ca^<2+> inhibitor of elicitor-stimulated OXB resulted in little induction of defense genes. lO.OXB stimulated phospholypase A_2 depending on a GTP binding protein. 11. OXB-related cytoplasmic aggregation was dependent on actin molecules with actin-binding proteins. 12. OXB did not necessarily associated with induction of enzymes related with phytoalexin production. 13. Further characterization of 0_2 generating NADPH oxidase was necessary for understanding OXB at molecular level of enzyme and gene. Less

植物对病原体或激发子产生诱导防御反应。超氧化物生成反应（氧化爆发：OXB）的快速激活在防御反应的诱导开始时被诱导。该反应系统依赖于细胞膜上的NADPH氧化酶，并受Ca^（2+）、钙调蛋白、蛋白激酶等信号转导系统的调控。本研究对NADPH氧化酶的分离、鉴定、OXB刺激的信号转导以及OXB诱导的防御相关基因的表达进行了研究。1.马铃薯块茎切片后老化过程中切片表面上OXB反应潜力的时程增加。2.在远离激发子处理侧的老化切片表面用局部OXB诱导亚系统OXB。马铃薯块茎离体质膜O_2产生NADPH氧化酶的动力学4.各种Ca^2+信号传导或蛋白激酶抑制剂可抑制细胞凋亡， ...更多信息 局部和亚系统的选择。5.去污剂对质膜产生NADPH氧化酶的活性O_2的溶解作用6.可溶性NADPH氧化酶电泳后聚丙烯酰胺凝胶中几条活性带的检测。7.人白细胞gp 91 p ^同源cDNA的克隆<phox>及其与水稻和拟南芥同源性的检测8.在<phax>马铃薯块茎组织可溶性蛋白的SDS-PAGE上的级分中，通过抗人白细胞p67 α的抗体检测信号。9.用Ca^&lt;2+&gt;抑制剂抑制激发子刺激的OXB几乎不诱导防御基因。10.OXB依赖GTP结合蛋白刺激磷脂酶A_2。11. OXB相关的细胞质聚集依赖于肌动蛋白结合蛋白的肌动蛋白分子。12. OXB不一定与植物抗毒素产生相关酶的诱导有关。13.对O_2产生NADPH氧化酶的进一步研究对于从酶和基因的分子水平上了解OXB是必要的。少

项目成果

Takemoto, D.: "Identification of chitinase and osmotin-like protein as actin-binding proteins in suspension-cultured potato cells." Plant Cell Physiol.38. 441-448 (1997)

Takemoto, D.：“在悬浮培养的马铃薯细胞中鉴定几丁质酶和渗透素样蛋白作为肌动蛋白结合蛋白。”

Senda, K.: "Effect of mastoparan on phospholipase A_2 activity in potato tubers treated with fungal elicitor." Plant Cell Physiol.38. 441-448 (1998)

Senda, K.：“mastoparan 对用真菌诱导子处理的马铃薯块茎中磷脂酶 A_2 活性的影响。”

Doke, N.: "The oxidative burst : roles in signal transduction and plant stress." In : Scandalious, J.G.(ed.) Oxidative Burst and the molecular Biology and Antioxidant Defense, Cold Spring Harbor Laboratory Press.785-813 (1997)

Doke, N.：“氧化爆发：在信号转导和植物应激中的作用。”

Senda,K.: "Effect of mastoparan on phospholipase A_2 activity in potato tubers treated with fungal elicitor." Plant Cell Physiol.39. 1080-1086 (1998)

Senda,K.：“mastoparan 对用真菌诱导子处理的马铃薯块茎中磷脂酶 A_2 活性的影响。”

道家紀志: "オキシダティブバーストと抵抗性発現.山田哲治・島本 功・渡辺裕一郎監修、分子レベルからみた植物の耐病性.(分担)" 秀潤社、東京., 196 (1998)

Noriyoshi Doike：“氧化爆发和抗性表达。从分子水平角度看植物抗病性，由 Tetsuji Yamada、Isao Shimamoto 和 Yuichiro Watanabe 监督。（共享）” Shujunsha，Tokyo.，196（1998）