To better understand the molecular mechanism of rice (Oryza sativa L.) in response to cold stress and further improve its tolerance to cold stress,a cold-responsive gene OsLCL3 was screened from cold stress expression profiles of rice gene chips.OsLCL3 was confirmed to be induced by low temperature (4℃) strongly with real-time fluorescent quantitative PCR and the expression of GUS reporter gene expression in stable transgenic rice driven by the promoter of OsLCL3.Homologous protein sequence analysis showed that OsLCL3 and its homologous proteins belong to DUF761 super family.Phylogenetic analysis of part members of this family including four homologs in rice genome was conducted.Subcellular localization analysis of OsLCL3 in rice protoplast showed that it was localized in nucleus and cytoplasm.Transgenic rice lines with over-expressing OsLCL3 were obtained and three of them were tested for low temperature stress tolerance.Results showed that the electrical conductivity of overexpression lines under the low temperature stress was significantly lower than that of wild type plants.The survival rate of overexpression lines was significantly higher than that of wild type plants.These results indicate that OsLCL3 may play an important role in low temperature response in rice.