Yeasts are known to colonize various natural and anthropogenic habitats, notably vineyards and wineries. The recent literature shows that there is a growing interest in indigenous yeast starters, reflective of the local yeast biodiversity and potentially adapted to cultivated grape varieties. This assumes that native yeast strains could be linked to the notion of “terroir”. Within this biodiversity, yeasts participate in complex microbial communities associated with distinct niches in the vineyards and wineries. This natural yeast biodiversity could represent a unique microbial signature of the “terroir” and an invaluable resource for winemakers. The present study revealed the diversity of yeast species in South-Western Switzerland. Comprehensive sampling was conducted across eight Swiss vineyards and their associated wineries. Yeasts were isolated from collected samples encompassing vineyard niches (flowers, vine bark, insects, grape berries) and winery environments (equipment surfaces, walls). Molecular identification was performed through sequencing of the D1/D2 domain of the 26S ribosomal DNA gene. The analysis identified 263 isolates, belonging to 44 species distributed across 25 distinct genera. Subsequently, a subset of these yeasts has undergone characterization in co-fermentations with S. cerevisiae reported in another work. Characterization studies, including micro-fermentations alongside chemical and sensory analyses, will be extended to additional species.