Molecular Regulation of Antioxidant Melatonin Biosynthesis by Brassinosteroid Acting as an Endogenous Elicitor of Melatonin Induction in Rice Seedlings

Gibberellic acid (GA) was recently shown to induce melatonin synthesis in rice. Here, we examined whether brassinosteroids (BRs) also induce melatonin synthesis because BRs and GA show redundancy in many functions. Among several plant hormones, exogenous BR treatment induced melatonin synthesis by twofold compared to control treatment, whereas ethylene, 6-benzylaminopurine (BA), and indole-3-acetic acid (IAA) showed negligible effects on melatonin synthesis. Correspondingly, BR treatment also induced a number of melatonin biosynthetic genes in conjunction with the suppression of melatonin catabolic gene expression. Several transgenic rice plants with downregulated BR biosynthesis-related genes, such as DWARF4, DWARF11, and RAV-Like1 (RAVL1), were generated and exhibited decreased melatonin synthesis, indicating that BRs act as endogenous elicitors of melatonin synthesis. Notably, treatment with either GA or BR fully restored melatonin synthesis in the presence of paclobutrazol, a GA biosynthesis inhibitor. Moreover, exogenous BR treatment partially restored melatonin synthesis in both RAVL1 and Gα RNAi transgenic rice plants, whereas GA treatment fully restored melatonin synthesis comparable to wild type in RAVL1 RNAi plants. Taken together, our results highlight a role of BR as an endogenous elicitor of melatonin synthesis in a GA-independent manner in rice plants.