Hydrogen peroxide acts as a second messenger for the induction of defense genes in tomato plants in response to wounding, systemin, and methyl jasmonate

The systemic accumulation of both hydrogen peroxide (H2O2) and proteinase i nhibitor proteins in tomato leaves in response to wounding was inhibited by the NADPH oxidase inhibitors diphenylene iodonium (DPI), imidazole, and py ridine. The expression of several defense genes in response to wounding, sy stemin, oligosaccharides, and methyl jasmonate also was inhibited by DPI. T hese genes, including those of four proteinase inhibitors and polyphenol ox idase, are expressed within 4 to 12 hr after wounding. However, DPI did not inhibit the wound-inducible expression of genes encoding prosystemin, lipo xygenase, and allene oxide synthase, which are associated with the octadeca noid signaling pathway and are expressed 0.5 to 2 hr after wounding. Accord ingly, treatment of plants with the H2O2-generating enzyme glucose oxidase plus glucose resulted in the induction of only the later-expressed defensiv e genes and not the early-expressed signaling-related genes. H2O2 was cytoc hemically detected in the cell walls of vascular parenchyma cells and spong y mesophyll cells within 4 hr after wounding of wild-type tomato leaves, bu t not earlier. The cumulative results suggest that active oxygen species ar e generated near cell walls of vascular bundle cells by oligogalacturonide fragments produced by wound-inducible polygalacturonase and that the result ing H2O2 acts as a second messenger for the activation of defense genes in mesophyll cells. These data provide a rationale for the sequential, coordin ated, and functional roles of systemin, jasmonic acid, oligogalacturonides, and H2O2 signals for systemic signaling in tomato plants in response to wo unding.