2-Hydroxymelatonin Induces Husk-Imposed Vivipary in the Transgenic Rice Overexpressing Melatonin 2-Hydroxylase

Pre-harvest sprouting (PHS) reduces the quality and quantity of crop seeds. PHS can be imposed through the embryo or husk pathway of cereal crops. Most reported PHS seeds are imposed via the embryo pathway. Here, we generated transgenic rice plants overexpressing rice melatonin 2-hydroxylase (<i>OsM2H</i>), which catalyzes the hydroxylation of melatonin to 2-hydroxymelatonin (2-OHM). <i>OsM2H</i> overexpression (M2H-OE) showed PHS under paddy conditions. Germination assays revealed that intact seeds harvested at 26 and 36 days after heading (DAH) showed PHS, whereas dehusked seeds did not, indicating husk-imposed PHS. Overproduction of 2-OHM was observed in M2H-OE seeds compared to wild-type control. In addition, M2H-OE lines produced more hydrogen peroxide than the wild-type. 2-OHM-induced reactive oxygen species resulted in the induction of <i>OsGA3ox2</i>, a gibberellin (GA) biosynthesis gene, and repression of <i>OsGA2ox3</i>, a GA degradation gene, in caryopses at 2 DAH, but in the induction of the ABA degradation gene <i>OsABA8ox3</i> in intact seeds at 26 DAH. In addition, M2H-OE seedlings were longer and showed increased levels of hydrogen peroxide and <i>OsGA3ox2</i> expression versus the wild-type. This is the first report showing that 2-OHM can induce PHS via the husk pathway in rice seeds through the induction of GA biosynthetic and ABA degradation genes.