Hydrogen peroxide regulated salicylic acid- and jasmonic acid-dependent systemic defenses in tomato seedlings

To investigate the regulation of hydrogen peroxide (H2O2) in SA- and JA-dependent systemic defense pathways in tomato seedlings damaged by cotton bollworm. The lower two leaves of four-leaf tomato plant seedlings were treated with cotton bollworms. The upper leaves were then removed at various time points and assessed for systemic changes in H2O2 levels, enzyme activity and defense-related genes expression. Wild-type seedlings, def-5 mutant seedlings (deficient in JA accumulation) and transgenic nahG seedlings(deficient in SA content) were constructed at four-leaf stage. Wild-type plants were further treated with dimethylthiourea (DMTU) and 2,5-dihydroxycinnamic acid methyl ester (DHC). Bollworm feeding in the lower leaves induced rapid systemic accumulation of H2O2 in the upper leaves. H2O2 accumulation further increased activities for SA- and JA-dependent enzymes including proteinase inhibitors (PIs), pathogenesis-related gene 1 (PR-1), cathepsin D inhibitor (CDI), beta-1,3-glucanse (BGL) and polyphenol oxidase (PPO). Furthermore, H2O2 accumulation enhanced mRNA expression of BGL-2, PR-1, PI, CDI and PPO genes. Suppression of H2O2 accumulation using H2O2 scavengers substantially diminished these effects. mRNA expression of PR-1 was not induced in transgenic nahG plants. SA- and JA-dependent signaling pathways are involved in the tomato systemic defense responses to herbivores, and that H2O2 generation is required for both systemic pathways.