Soil salinity is a threat to agricultural production worldwide. Oat (Avena sativa L.) is an irreplaceable crop in areas with fragile ecological conditions. However, there is a lack of research on salt tolerance evaluation of oat germplasm resources. Therefore, the purpose of this work was to evaluate the salt tolerance of oat cultivars and investigate the mechanism of salt-tolerant oat cultivars’ adaptation to salinity. Salt tolerance of 100 oat cultivars was evaluated, and then two salt-tolerant cultivars and two salt-sensitive cultivars were used to compare their physiological responses and expression patterns of Na+- and K+-transport-related genes under salinity. Principal component analysis and membership function analysis had good predictability for salt tolerance evaluation of oat and other crops. The 100 oat cultivars were clustered into three categories, with three salt tolerance levels. Under saline condition, salt-tolerant cultivars maintained higher growth rate, leaf cell membrane integrity, and osmotic adjustment capability via enhancing the activities of antioxidant enzymes and accumulating more osmotic regulators. Furthermore, salt-tolerant cultivars had stronger capability to restrict root Na + uptake through reducing AsAKT1 and AsHKT2;1 expression, exclude more Na+ from root through increasing AsSOS1 expression, compartmentalize more Na + into root vacuoles through increasing AsNHX1 and AsVATP-P1 expression, and absorb more K+ through increasing AsKUP1 expression, compared with salt-sensitive cultivars. The evaluation procedure developed in this work can be applied for screening cereal crop cultivars with higher salt tolerance, and the elucidated mechanism of oat adaptation to salinity lays a foundation for identifying more functional genes related to salt tolerance.