【Objective】 In order to determine the relatively strong adaptability of alfalfa(Medicago sativa) varieties under flooding stress.【Method】 The pot covering method was used to simulate the flooding conditions and explore the impact on the biological characteristics of alfalfa seedlings,in order to provide basic materials for the study of alfalfa waterlogging tolerance.【Result】 The results showed that the plant height of alfalfa was higher than that of control after 7 days of flooding. The stem dry weight,leaf dry weight and aboveground dry biomass of Juneng 7,gladiator,Juneng 801,WL440HQ,WL525HQ and WL656HQ were higher than those of control. The main root length of Juneng 995,gladiator,Juneng 801,WL525HQ,WL656HQ and WL712 were higher than that of control. At the 14th day of flooding stress,only 2 ‐3 varieties had higher main root length,stem dry weight,leaf dry weight and aboveground dry biomass than the control.【Conclusion】 The comprehensive evaluation of waterlogging tolerance was carried out by principal component analysis and subordinate function method. The waterlogging tolerance of Juneng 995, WL440HQ,gladiator and other varieties was higher.