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

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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.