Study on physiological and ecological responses of alfalfa during the germination stage under drought, bicarbonate, and combined stress conditions

Summary Alfalfa (Medicago sativa L.) is frequently constrained by drought and salinity during cultivation and production. This study aimed to investigate the differential response characteristics and underlying physiological and ecological mechanisms of alfalfa during the germination stage under drought stress, bicarbonate stress, and their combined effects. Twelve alfalfa varieties were subjected to germination tests using PEG-6000 to simulate drought stress (0–20%) and NaHCO□ to simulate bicarbonate stress (0–30 mM). Based on the semi-inhibitory concentrations and phenotypic indices observed under single-stress conditions, a combined stress treatment was designed. The subordinate function method was employed to comprehensively evaluate the variations in stress responses among the twelve alfalfa varieties during the germination stage. Additionally, this study explored the physiological and ecological response mechanisms of drought-tolerant, salt-tolerant, and drought-salt-sensitive varieties. Among the 12 alfalfa varieties, WL363HQ has the strongest drought and salt tolerance, while WL319HQ is the most sensitive to drought and salt stress. Based on this, the varieties can be classified into drought-tolerant, salt-tolerant, and sensitive categories. At low concentrations, the drought-salt combination shows antagonism (salt alleviates drought inhibition); at high concentrations, it turns into synergy, and the inhibitory effect is amplified.WL363HQ adapts to the combined stress by maintaining stable SOD activity and gradually accumulating soluble sugar; WL319HQ shows sensitivity due to fluctuating SOD, membrane lipid peroxidation, and uncontrolled soluble sugar. Competing Interest Statement The authors have declared no competing interest.