Integrating Physiology, Transcriptome, and Metabolomics Reveals the Potential Mechanism of Nitric Oxide Concentration-Dependent Regulation of Embryo Germination in .

Nitric oxide (NO) breaks a seed's dormancy and stimulates germination by signaling. However, the key physiological metabolic pathways and molecular regulatory mechanisms are still unclear. Therefore, this study used physiological, transcriptomic, and metabolomics methods to analyze the key genes and metabolites involved in the NO regulation of plant embryo germination and their potential regulatory mechanisms. The physiological analysis results indicate that the appropriate concentration of NO increased the content of NO and hydrogen peroxide (HO) in cells, stimulated the synthesis of ethylene and jasmonic acid (JA), induced a decrease in abscisic acid (ABA) content, antagonistic to the gibberellin (GA) effect, and promoted embryo germination and subsequent seedling growth. However, the high concentrations of NO caused excessive accumulation of HO, destroyed the reactive oxygen species (ROS) balance, and inhibited embryo germination and seedling growth. The combined analysis of transcriptomics and metabolomics showed that the genes related to phenylpropanoid (phenylalanine ammonia-lyase, trans-cinnamate 4-monooxygenase, ferulate-5-hydroxylase, coniferyl-alcohol glucosyltransferase), and flavonoid synthesis (10 genes such as ) were significantly up-regulated during embryo germination. The high concentration of exogenous NO inhibited embryo germination by up-regulating the expression of 4-coumaric acid coenzyme A ligase (4CL) and negatively regulating the expression of flavonoid synthesis genes. This suggests that NO concentration-dependently regulates phenylpropanoid and flavonoid biosynthesis, thereby affecting ROS metabolism and hormone levels, and ultimately regulates the dormancy and germination of embryos.