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

研究了马铃薯（Solanum tuberosum L.）块茎组织和大豆（Phaseolus vulgaris L.）下胚轴组织中菌丝壁（HWC）激发子和抑制子（Mr. 4700和Mr. 440）的体外蛋白磷酸化作用。磷酸化水平估计为25 muM游离Ca^<2+>。为了测定^<32> p并入[H^+] atp酶，磷酸化蛋白通过SDS-PAGE分离。^<32>蛋白带P水平用ANBIS放射分析成像系统分析。抑制因子（250马克杯/毫升）比激发因子（250马克杯/毫升）更能刺激马铃薯质膜[H^+] atp酶的磷酸化。抑制因子还能刺激豆质膜atp酶活性。水杨酸（15 ~ 30 muM）刺激了马铃薯和豆类的磷酸化水平。相反，过氧化氢降低了两种植物的蛋白磷酸化水平。水杨酸和过氧化氢是植物防御的潜在化学信号。我们得出结论，特定的蛋白激酶(s)影响p型[H^+] atp酶。真菌病原体成分对蛋白激酶活性的调节可能参与植物防御机制的膜信号转导。采用细胞悬浮介质化学发光法测定了大豆悬浮细胞中活性氧基团（超氧自由基和过氧化氢）的产生。菌丝壁激发剂引起活性氧基团的产生。相比之下，真菌的抑制因子对产量没有刺激作用。菌丝壁激发剂（250马克杯/毫升）在15分钟内使马铃薯悬浮细胞产生活性氧和过氧化氢，但真菌的葡聚糖使其产生的活性氧和过氧化氢少于激发剂。

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.