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ANALYSIS OF THE REGULATION GENE FOR HYPERSENSITIIVITY OF PLANT AND ANALYSIS OF SIGNAL TRUNSDUCTION IN PLANT CELL

植物过敏调控基因分析及植物细胞信号转导分析

基本信息

批准号：
05660047
负责人：
FURUICHI Naotaka
金额：
$ 1.34万
依托单位：
NIIGATA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

In vitro protein phosphorylation of the plasma membrane proteins of potato (Solanum tuberosum L.) tuber tissue and bean (Phaseolus vulgaris L.) hypocotyl tissue was stimulated by an hyphal wall (HWC) elicitor and suppressor (Mr. 4,700 and Mr 440) isolated from Phytophthora infestans. Phosphorylation levels were estimated at 25 muM free Ca^<2+>. For determination of ^<32>P-incorporation into [H^+] ATPase, phosphorylated proteins were separated by SDS-PAGE.^<32>P levels in the protein bands were analyzed by the ANBIS Radioanalytic Imaging System. The suppressor (250 mug/ml) stimulated phosphorylation of the plasma membrane [H^+]ATPase in potato more than the elicitor (250 mug/ml). Suppressor also stimulated the ATPase activity of bean plasma membranes.Salicylic acid (15-30 muM) stimulated the phosphorylation level of potato and bean. In contrast, hydrogen peroxide decreased the protein phosphorylation level of both plants. Salicylic acid and hydrogen peroxide are potential chemical signals in plant defenses. We conclude that specific protein kinase(s) affect the P-type [H^+] ATPase. Regulation of protein kinase activity by fungal pathogen components may participate in membrane signal transduction for plant defense mechanisms.Production of active oxygen group (superoxide radicals and hydrogen peroxide) in bean suspension cells were measured by the detectioin of chemiluminescence in cell suspension media. Hyphal wall elicitor caused genaration of active oxigen groups. In contrast, the suppressor of the fungus did not stimulate the production.Hyphal wall elicitor (250 mug/ml) caused the production of active oxygen and hydrogen peroxide in potato suspension cells within 15 min, but the glucan of the fungus caused its production less than elicitor.

马铃薯质膜蛋白的体外蛋白质磷酸化块茎组织和菜豆(Phaseolus Common garis L.)下胚轴组织被从致病疫霉(Phytophthora Infestans)分离的菌丝壁(HWC)激发子和抑制子(MR 4700和MR 440)刺激。磷酸化水平估计为25微米的游离钙离子。为了确定[H^+]ATPase中^&lt；32&gt；P的掺入量，用SDS-PAGE分离磷酸化蛋白，用ANBIS放射分析成像系统分析蛋白条带中^&lt；32&gt；P的含量。抑制物(250µg/ml)对马铃薯质膜[H~+]ATPase磷酸化的促进作用强于激发子(250µg/ml)。水杨酸(15-30 mU)对马铃薯和菜豆的磷酸化水平有促进作用。相反，过氧化氢降低了两种植物的蛋白质磷酸化水平。水杨酸和过氧化氢是植物防御中潜在的化学信号。我们的结论是，特定的蛋白激酶(S)影响P型[H^+]ATPase。病原菌成分对蛋白激酶活性的调节可能参与了植物防御机制的膜信号转导。通过细胞悬浮液中化学发光的检测，测定了大豆悬浮细胞中活性氧自由基(超氧阴离子自由基和过氧化氢)的产生。菌丝壁激发子引起活性氧基团的生成。菌壁激发子(250µg/ml)能在15min内诱导马铃薯悬浮细胞产生活性氧和过氧化氢，而菌体中的葡聚糖对马铃薯悬浮细胞产生的过氧化氢和活性氧的影响小于激发子。