Stripe rust caused wheat yields losses every year, which was a great threat to wheat grain security. TGA factors have been confirmed to form complex with SA receptor NPR1, and they were considered to participate in the plant defense, but the study about TGA transcription factors function in wheat was limited. In this study, we identified 38 TGA transcription factors in wheat genome. Through phylogenetic analysis, the distance of these TaTGAs with other plants was figured out, which includes its two progenitors and two well studied model plant. On the basis of that, the conserved motifs of these TGAs were analysed to infer the function approximately. Then we screened the roles of TaTGAs in biotic stress from transcriptome data and acquired some TaTGAs had important roles in pathogen resistance. Subsequently, some TaTGAs with significantly differential expression were confirmed by qRT-PCR. The results showed the transcriptional levels of some TaTGAs can up to 7 times higher after treatment with salicylic acid or pathogen infection than the contrast. In addition to this, some ROS-response genes show the same changes. Subcellular location and transcriptional activation assay showed the TaTGA13 located in the nucleus and possessed transactivate ability. In the end, Virus-induced gene co-silencing led to downregulation of TaTGA13, expressing enhanced resistance to stripe rust in phenotypes. The phenotype of the TGAs mutation with stripe rust inoculation confirmed results of VIGS assay, it showed fungal growth was slightly decreased, and the ROS-response genes were reduced in the mutant. These results suggest that TaTGAs play an important role in wheat stripe rust resistance. It provides a theoretical basis for wheat stripe rust resistance breeding.