Abstract: The gene family of the WRKY transcription factors in plants play an important role in controlling leaf senescence. The WRKY53 gene is closely related to leaf senescence. Based on the EST sequence, the full-length CDS of SpWRKY53 was cloned from Stipa purpurea. Sequence analysis showed that the opening reading frame of SpWRKY53 was 1443 bp, which encoded a 480-amino acid peptide. Its protein molecular weight and isoelectric point were 50.9 kD and 7.64, respectively. Sequence alignment and phylogenetic analysis results indicated that the SpWRKY53 gene with two WRKY sequences displayed similar structures with those reported for the WRKY53 gene of wheat. SpWRKY53 was localized in the cell nucleus by subcellular analysis. SpWRKY53 was upregulated and could promote leaf senescence under abiotic stress. This study will lay a foundation for further study of the WRKY53gene in the molecular breeding of forage grass.