Exogenous melatonin promotes the growth of alfalfa (Medicago sativa L.) under NaCl stress through multiple pathways.

Salinity is one of the most common factors affecting alfalfa (Medicago sativa L.), and NaCl is one of the main factors of salinity stress which can cause heavy losses in agricultural production in the world. The application of exogenous melatonin (MT) plays a major role in numerous plants against various stress environments. The effects of exogenous MT on the NaCl tolerance of alfalfa treated with the control, 100 µmol L-1 MT, 150 mmol L-1 NaCl, or 150 mmol L-1 NaCl+ 100 µmol L-1 MT were investigated. The results showed that MT increased growth parameters, inhibited chlorophyll degradation and promoted photosynthetic gas exchange parameters (photosynthetic rate, conductance to H2O, and transpiration rate) and stomatal opening under NaCl stress. Osmotic regulation substances such as soluble sugar, proline and glycine betaine were the highest in the NaCl treatment and the second in the NaCl+MT treatment. Nitrogen, phosphorus, potassium, calcium and magnesium were reduced and sodium was increased by NaCl, whereas these levels were reversed by the NaCl+MT treatment. MT inhibited cell membrane imperfection, lipid peroxidation and reactive oxygen species (ROS) accumulation caused by NaCl stress. MT up-regulated the gene expression and activity of antioxidant enzymes and increased the content of antioxidant non-enzyme substances to scavenge excessive ROS in NaCl-treated plants. In addition, all indicators interacted with each other to a certain extent and could be grouped according to the relative values. All variables were divided into PC 1 (89.2 %) and PC 2 (4 %). They were clustered into two categories with opposite effects, and most of them were significant variables. Hence, these findings reveal that exogenous MT alleviates the inhibitory effects of NaCl stress on photosynthesis, stomata opening, osmotic adjustment, ion balance and redox homeostasis, enhancing tolerance and growth of alfalfa. Furthermore, it suggests that MT could be implemented to improve the NaCl tolerance of alfalfa.