In order to reveal the roles of asparagus lettuce WRKYⅢ subfamily transcription factors in different biological processes, we cloned two WRKYⅢ subfamily transcription factor genes LsWRKY08 and LsWRKY37 from asparagus lettuce cultivar ‘Yonganhong’ in the study. The sequence alignment, evolutionary tree construction, qRTPCR, interaction network and promoter analysis were carried out. The results will provide a basis for further studying the function of WRKYⅢ transcription factor of asparagus lettuce and improving the yield and quality of asparagus lettuce. The results showed that: (1) LsWRKY08 and LsWRKY37 transcription factors contained 945 bp and 930 bp open reading frames, encoding 314 and 309 amino acid residues, respectively. (2) qRTPCR analysis showed that the expression of LsWRKY08 in leaves was higher than that in roots and stems; in the process of stem expansion, its expression decreased gradually; but compared with control, the expression patterns of LsWRKY08 increased in drought, high/low temperature and salt; the expression of LsWRKY08 increased significantly in SA treatment compared with control. LsWRKY37 showed a higher level of expression in roots than in leaf and stem. Its expression pattern during stem expansion was different to that of LsWRKY08. There were differences in the expression pattern of LsWRKY37 under different abiotic stresses. (3) Interaction network analysis showed that LsWRKY08 and LsWRKY37 could interact with plant defense related proteins and other transcription factors. Promoter analysis showed the promoter regions of LsWRKY08 and LsWRKY37 genes contained multiple cis elements. Wbox element was identified in the promoter region of LsWRKY37, which indicating the selfregulation and crossregulation existed between LsWRKY37 and other WRKY transcription factors. These results showed that LsWRKY08 and LsWRKY37 genes could respond to different abiotic stresses and hormone treatments, but the expression patterns of these two genes were different, which indicating that the function of transcription factor genes belonged to the same subfamily may be different. LsWRKY08 and LsWRKY37 transcription factors could participate in different biological processes through interaction with other proteins or regulation by other genes.