The reduction of phytochemicals applied to grapevine relies on the development of alternative strategies involving activation of the plant's own defense system. The aim of this work was to study the signaling of defense responses to pathogens in Vitis vinifera. We identified in V. vinifera cv. Chardonnay two putative regulatory elements, VvNHL1 and VvEDS1, with similarity to Arabidopsis defense regulators NDR1 and EDS1. Expression studies of these putative signaling genes together with other known grape defense genes show that they are differentially regulated by salicylic acid and jasmonate-ethylene treatments, as well as by inoculation with different types of pathogens. The expression of VvEDS1 was stimulated by salicylic acid treatment, Botrytis cinerea and Plasmopara viticola inoculation, whereas VvNHL1 was repressed by B. cinerea. VvNHL1 overexpression introduced in Arabidopsis ndr1 mutant did not complement the mutation in terms of sensitivity to avirulent Pseudomonas syringae pv. tomato. Moreover, we observed a weakened resistance to B. cinerea of ndr1 mutants overexpressing VvNHL1, which may be related to cell death enhancement. Together, our results identify two new pathogen-responsive regulatory elements in Vitis vinifera, with potential roles in pathogen defense.