Metabolomic and physiological analysis of alfalfa (Medicago sativa L.) in response to saline and alkaline stress

Alfalfa (Medicago sativa L.) is a leguminous forage widely grown worldwide. Saline and alkaline stress can affect its development and yield. To elucidate the physiological mechanisms of alfalfa in response to saline and alkaline stress, we investigated the growth and physiological and metabolomic changes in alfalfa under saline (100 mM NaCl) and alkaline (100 mM Na2CO3, NaHCO3) stress. At the same Na+ concentration, alkaline stress caused more damage than that caused by saline stress. A total of 65 and 124 metabolites were identified in response to saline and alkaline stress, respectively. Determination of gene expression, enzyme activity, substance content, and KEGG enrichment analysis in key pathways revealed that alfalfa responded to saline stress primarily by osmoregulation and TCA cycle enhancement. Flavonoid synthesis, TCA cycle, glutamate anabolism, jasmonate synthesis, and cell wall component synthesis increased as responses to alkaline stress. This study provides important resources for breeding saline–alkaline-resistant alfalfa.