Abstract: A forward subtractive cDNA library was constructed from tall fescue “Houndog 5” cultivar exposed to low temperature stress via suppression subtractive hybridization (SSH). Then cold-resistance of tall fescue (induced by low temperaturing) was determined through genes expression analysis. A total of 36 ESTs from SSH-cDNA library were selected and sequenced after reverse northern hybridization screening. The sequencing results were analyzed by means of retrieval and alignment in unigene GeneBank database. Based on the analysis, 34 genes exhibited different expressions, of which the functions of 33 genes were known and identified. These functionally known genes were detected in signal transduction, photosynthesis, sugar metabolism, material transportation and stress resistance.