Heat stress is one of the major abiotic factors limiting the growth of cool-season grass species during summer season. The objectives of this study were to assess genetic variations in the transcript levels of selected genes in fine fescue cultivars differing in heat tolerance， and to identify single nucleotide polymorphism (SNP) markers associated with candidate genes related to heat tolerance. Plants of 26 cultivars of five fine fescue species ( spp.) were subjected to heat stress (38/33 °C， day/night temperature) in controlled environmental growth chambers. Physiological analysis including leaf chlorophyll content， photochemical efficiency， and electrolyte leakage demonstrated significant genetic variations in heat tolerance among fine fescue cultivars. The transcript levels of selected genes involved in photosynthesis (RuBisCO activase， Photosystem II CP47 reaction center protein)， carbohydrate metabolism (Sucrose synthase)， energy production (ATP synthase)， growth regulation (Actin)， oxidative response (Catalase)， and stress protection (Heat shock protein 90) were positively correlated with the physiological traits for heat tolerance. SNP markers for those candidate genes exhibited heterozygosity， which could also separate heat-sensitive and heat-tolerant cultivars into clusters. The development of SNP markers for candidate genes in heat tolerance may allow marker-assisted breeding for the development of new heat-tolerant cultivars in fine fescue and other cool-season grass species.