BackgroundThe myeloblastosis (MYB) superfamily is the largest transcription factor family in plants that play diverse roles during stress responses. However, the biotic stress-responsive MYB transcription factors of the grapevine have not been systematically studied. In China, grapevine berries are often infected with the grapevine berry inner necrosis virus (GINV), which eventually reduces the nutritional quality and commodity value.ResultsThe present study identified and characterized 265VvMYBorVvMYB-related genes of the “Crimson seedless” grapevine. Based on DNA-binding domain analysis, these VvMYB proteins were classified into four subfamilies, including MYB-related, 2R-MYB, 3R-MYB, and 4R-MYB. Phylogenetic analysis divided the MYB transcription factors into 26 subgroups. Overexpression ofVvMYB58suppressed GINV abundance in the grapevine. Further qPCR indicated that among 41 randomly selectedVvMYBgenes, 12 were induced during GINV infection, while 28 were downregulated. These findings suggest thatVvMYBgenes actively regulate defense response in the grapevine.ConclusionA deeper understanding of the MYB TFs engaged in GINV defense response will help devise better management strategies. The present study also provides a foundation for further research on the functions of the MYB transcription factors.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-023-04296-7.