Requirement of Host Signaling Mechanisms for the Action of Ptr ToxA in Wheat
Ptr ToxA, the host-selective toxin produced by Pyrenophora tritici-repentis, is genetically associated with the development of tan spot disease of wheat. The toxin was shown previously to cause a programmed cell death in the host that requires de novo mRNA and protein synthesis. In the present study, inhibitors of plant signaling mechanisms protected wheat leaves from toxin action, as determined by electrolyte leakage bioassays, when applied to leaves with toxin. Okadaic acid, calyculin A and phenylarsine oxide, all inhibitors of protein phosphatase activity, reduced toxin-induced electrolyte leakage by more than 90%. Inorganic calcium channel blockers (LaCl3 and CoCl2 reduced toxin-induced electrolyte leakage by 78–95%, depending on inhibitor and time of measurement. By comparison, about 50% protection was achieved by the application of the protein kinase inhibitors staurosporine and K-252A. Nonetheless, the reduction in toxin-induced electrolyte leakage by protein kinase inhibitors was reproduced in multiple trials and was statistically significant. The data indicate that host signaling mechanisms, including calcium fluxes and a protein phosphorylation cascade, are required for the Ptr ToxA-induced cell death in wheat. Our current model holds that the signaling events occur between toxin perception by the cell and the toxin-directed gene expression in the host associated with cell death. As an alternative, the toxin-induced mRNA synthesis required for cell death may be for protein phosphatase and/or protein kinase genes. Additional work is required to resolve these possibilities.
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