For a better understanding of stress responses of desert plant Ammopiptanthus mongolicus, we reported molecular characterization and expression analysis of AmLEA14 gene from the EST data of cold and droughtstressed A. mongolicus. Sequence analysis showed that the overall length of gene AmLEA14 was 579 bp, and the cDNA of AmLEA14 contained a 459 bp ORF encoding a polypeptide of 152 amio acids with a calculated molecular mass of 166 kDa and a theoretical pI of 49. Sequence and structural analysis showed that AmLEA14 protein was a member of late embryogenesis abundant protein class 4. Phylogenetic analysis indicated that AmLEA14 protein was most closely related to soybean LEA14 protein (P465191). Realtime quantitative PCR analysis showed that AmLEA14 was constitutively expressed, upregulated by cold, drought stressed, and specifically accumulated at the late stage of cold treatment. Otherwise, AmLEA14 was downregulated by heat stress. The combined results indicate that AmLEA14 protein plays some role in abiotic stress responses, mainly in cold tolerant mechanism, and is valuable for future abiotic stress researches in A. mongolicus.