WRKY transcription factors are widely involved in plant growth and development and play important roles in biotic and abiotic stress responses. In this study we cloned the EnWRKY26 gene from Elymus nutans Griseb ‘Ba Qing’ and conducted bioinformatics analysis， subcellular localization analysis， gene expression profiling， and transcription factor activity validation. The results indicated that EnWRKY26 was 996 bp in length， encoding 331 amino acids， with a molecular weight of 35 501.70 Da. The theoretical isoelectric point of the EnWRKY26 protein was 9.49， with an instability index of 53.78 and an average hydrophobicity of -0.652， indicating that it was an unstable hydrophilic protein. The entire protein sequence lacks transmembrane domains， and its secondary structure is predominantly composed of random coils. EnWRKY26 is most closely related to TraseCS7B02G418600 and TraseCS7D02G497700 in terms of phylogenetic relationship. Subcellular localization analysis revealed that EnWRKY26 was localized in the nucleus. Transcription factor activity validation confirmed that it possesses transcriptional activation activity. Under salt stress treatment， the relative expression level of EnWRKY26 showed an increasing trend. This study provides a theoretical basis for an in-depth understanding of the molecular mechanism by which EnWRKY26 regulates the response of Elymus nutans to salt stress.

Elymus nutans  /  EnWRKY26  /  Gene cloning  /  Expression analysis  /  Transcription factors