This study aimed to develop and use a culturomic approach to build a collection of grapevine foliar microorganisms with potential applications for the biocontrol of grapevine foliar diseases, particularly downy mildew (caused by the oomycete Plasmopara viticola). A custom grapevine leaf-based culture medium (GLA) was developed and tested to enhance the cultivability of microbial communities associated with grapevine leaf surface (phyllosphere) and leaf endosphere. This plant-based medium was coupled with conventional diluted and undiluted laboratory media to isolate bacteria, yeast and filamentous fungi from grapevine leaves collected from vineyards in the Médoc region of Bordeaux in spring 2022. Isolates were identified by combining Matrix-Assisted Laser Desorption/Ionisation Time Of Flight Mass Spectroscopy (MALDI-TOF MS) and Sanger sequencing. The dataverse contains the input files and R scripts used for all statistical analyses carried out in the study.