Abstract BACKGROUND Powdery mildew in grapevine is caused by Erysiphe necator and its control requires many chemical treatments. Numerous efforts are being made to improve disease management to achieve crop sustainability goals. The exogenous induction of plant immune responses is one of the most encouraging strategies currently being developed. The objective of this research was to analyse differences in phenolic compound concentrations in E. necator ‐infected leaves of two varieties of Vitis vinifera , Tempranillo and Tempranillo Blanco, using ultra performance liquid chromatography coupled with mass spectrometry. To understand the susceptibility of the varieties, in vitro assays using whole leaves were done. RESULTS Differences in susceptibility between varieties were found in the early stage of the disease. In both varieties, total phenolic compounds were higher in infected leaves; however, hydroxycinnamic acid, anthocyanins and stilbenes were higher only in Tempranillo. Twenty‐six compounds showed differential responses to the fungal disease in Tempranillo, but only two in Tempranillo Blanco: syringa resinol, which was not detected in diseased leaves; and gallocatechin, which increased at 5 days post inoculation. In Tempranillo, four anthocyanidins, six hydroxycinnamic acids, mainly feruloyl derivates, and epigallocatechin gallate were higher in infected leaves at the beginning of the infection, whereas (−)‐epicatechin and protocatechuic hexoside contents were lower. CONCLUSION Disease‐induced changes in phenolic compound biosynthesis were found. The increase in anthocyanidin content and flavan‐3‐ol galloylation could have a role in delaying E. necator growth in Tempranillo. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.