Yeast community involved in plant–bumblebee mutualism was investigated in three successive years. Yeasts were isolated from floral nectar, bumblebee queens after hibernation, bumblebee workers, and the honey provisions in nests. From the distribution of yeast species in the various microhabitats in the course of the year their ecology was assessed. Nectar of numerous plant species belonging to various plant families was analyzed in order to uncover possible impacts on the yeasts present in the nectar. Only ascomycetous yeasts were autochthonous members of the communities in the plant–bumblebee mutualism. Species in the Metschnikowia clade, the Starmarella clade, and the genera Debaryomyces and Zygosaccharomyces were associated with the mutualism. Some species appeared highly specialized, whereas others had a broader distribution. While physical and chemical properties of nectar had only limited influence on the abundance of nectar yeasts, the attractiveness of plants to the flower-visiting insects appears to have had a greater impact on the abundance and frequency of yeasts in the nectar of different plant species.