Setaria viridis (green foxtail millet), a short life-cycle C4 plant in the Gramineae, serves as a resilient crop that provides good yield even in dry and marginal land. Although S. viridis has been studied extensively in the last decade, its defense responses, in particular the chemical defensive metabolites that protect it against insect herbivory, are unstudied. To characterize S. viridis defense responses, we conducted transcriptomic and metabolomic assays of plants infested with aphids and caterpillars. Pathway enrichment analysis indicated massive transcriptomic changes that involve genes from amino acid biosynthesis and degradation, secondary metabolites and phytohormone biosynthesis. The Trp-derived metabolite serotonin was notably induced by insect feeding. Through comparisons with known rice serotonin biosynthetic genes, we identified several predicted S. viridis Trp decarboxylases and cytochrome P450 genes that were up-regulated in response to insect feeding. The function of one Trp decarboxylase was validated by ectopic expression and detection of tryptamine accumulation in Nicotiana tabacum. To validate the defensive properties of serotonin, we used an artificial diet assay to show reduced Rhopalosiphum padi aphid survival with increasing serotonin concentrations. This demonstrated that serotonin is a defensive metabolite in S. viridis and is fundamental for understanding the adaptation of it to biotic stresses.

Characterizing the serotonin biosynthesis pathway upon aphid infestation in Setaria viridis leaves