Melatonin and dopamine mediate the regulation of nitrogen uptake and metabolism at low ammonium levels in Malus hupehensis.

In plants, ammonium (NH4+) is the main nitrogen source and acts as a physiological and morphological response signaling molecule. Melatonin and dopamine are associated with plant responses to abiotic stress. However, previous studies have rarely focused on nutrient stress, and the roles of melatonin and dopamine in the uptake and metabolism of nitrogen in plants remain unclear. In this study, we investigated the regulatory effects of melatonin and dopamine on nitrogen utilization efficiency in apple seedlings under two NH4+ concentrations (2 and 0.1 mM) by measuring plant growth, root system architecture, 15NH4+ content, and related enzyme activity and gene expression. Under low nitrogen supply, apple seedling growth slowed and showed marked reductions in biomass accumulation, chlorophyll content, and nutrient uptake. However, both melatonin and dopamine significantly improved plant growth, chlorophyll content, and root development and enhanced antioxidant enzyme activity. Exogenous application of melatonin or dopamine also promoted the absorption and accumulation of 15NH4+ and enhanced nitrogen metabolism-related enzyme activity. At the molecular level, melatonin and dopamine significantly increased the expression levels of nitrogen metabolism genes and transporter genes. Overall, these results suggest that melatonin and dopamine can relieve nutrient stress caused by low concentrations of NH4+ through regulating the absorption and metabolism of nitrogen.

Beneficial Effects of Exogenous Melatonin and Dopamine on Low Nitrate Stress in Malus hupehensis.

Malus hupehensis, as an apple rootstock, is an economically important tree species popular due to its excellent fruit yield and stress resistance. Nitrogen is one of the critical limiting factors of plant growth and fruit yield, so it is crucial to explore new methods to improve nitrogen use efficiency. Melatonin and dopamine, as multifunctional metabolites, play numerous physiological roles in plants. We analyzed the effects of exogenous melatonin and dopamine treatments on the growth, root system architecture, nitrogen absorption, and metabolism of M. hupehensis when seedlings were exposed to nitrate-deficient conditions. Under low nitrate stress, plant growth slowed, and chlorophyll contents and 15NO3 - accumulation decreased significantly. However, the application of 0.1 µmol/L melatonin or 100 µmol/L exogenous dopamine significantly reduced the inhibition attributable to low nitrate levels during the ensuing period of stress treatment, and the effect of dopamine was more obvious. In addition to modifying the root system architecture of nitrate-deficient plants, exogenous melatonin and dopamine also changed the uptake, transport, and distribution of 15NO3 -. Furthermore, both exogenous melatonin and dopamine enhanced tolerance to low nitrate stress by maintaining the activity of enzymes (NR, NiR, GS, Fd-GOGAT, and NADH-GOGAT) and the transcription levels of related genes involved in leaf and root nitrogen metabolism. We also found that exogenous melatonin and dopamine promoted the expression of nitrate transporter genes (NRT1.1, NRT2.4, NRT2.5, and NRT2.7) in nitrate-deficient plant leaves and roots. Our results suggest that both exogenous melatonin and dopamine can mitigate low nitrate stress by changing the root system architecture, promoting the absorption of nitrate, and regulating the expression of genes related to nitrogen transport and metabolism. However, according to a comprehensive analysis of the results, exogenous dopamine plays a more significant role than melatonin in improving plant nitrogen use efficiency.