Sour rot in grapevines is thought to result from berry infection by yeasts, acetic acid bacteria and vinegar flies. To better characterize the role of each of the actors involved in sour rot expression, we conducted experiments involving the isolation of 1593 fungi and bacteria to understand the composition and dynamics of the microbiomes associated with healthy berries, diseased berries and insect vectors. As some grape clusters became symptomatic for sour rot in the absence of acetic acid bacteria, the latter might not necessarily be needed for disease expression. Similar to other yeast genera, the yeast genus Geotrichum is here also reported for the first time to be able to initiate sour rot in grapes, however, this finding has to be confirmed by further studies. By allowing or denying the access of insect vectors to intact or artificially wounded grapes we could emphasize that they accelerate the expression of the disease when berries are injured. Moreover, the microbial communities identified on native vinegar flies (Drosophila spp.) and the introduced Drosophila suzukii were similar and yeast dominated. This highlights the key role of these insect vectors in the transmission of microorganisms inflicting sour rot to wounded berries. Finally, our data also suggest that sour rot and grey mould can coexist in a vineyard at an advanced stage of grape decomposition, which runs counter to recent studies that emphasize the supremacy of sour rot over grey mould.