Drought stress is an important factor affecting wheat yield. In recent years, physiological phenomena related to glutamine synthase (GS) activity and drought resistance ability of wheat have been gradually discovered. However, the drought resistance function of GS gene in wheat has not been systematically identified. The GS family proteins of plants and microorganisms were studied in this study. Based on the whole genome database of common wheat, the TaGSs genes were systematically identified and its characteristics were analyzed by bioinformatics method. The result shows that fifteen TaGSs proteins were screened from the whole genome database of common wheat, which belonged to five subfamilies. TaGS genes in the same subfamily share high similarity in conserved motifs, intron-exon number, length and position. Combined with the transcriptome data of Qingmai 6, the expression of TaGSs in wheat leaves under abiotic stress was studied. The results showed that TaGSs genes showed different expression patterns in response to drought and salt stress. However, it was consistent that Ta4A/4B/4D.GSe were up-regulated in both abiotic stresses, and Ta4D.GSe was the most significantly up-regulated. Ta4D.GSe overexpression vector was constructed, and transgenic wheat lines have been obtained. The wilting degree of overexpressed wheat lines was significantly lower than that of the recipient wheat under natural drought treatment. The scavenging ability of reactive oxygen species was improved, the degree of lipid peroxidation of plant cell membrane was lower, and the content of proline and soluble sugar were significantly increased. These results indicated that overexpression of Ta4D.GSe could improve osmotic regulation ability of wheat, which laid a foundation for further study of drought resistance mechanism of Ta4D.GSe gene.