This study aimed to add diversity to the food fermented by yeasts. For this purpose, we isolated only the strains of Saccharomyces spp. and examined their several characteristics essential for fermented food. Furthermore, we tried to make bread using the strains selected. Two hundred and sixty-two strains of budding yeasts were isolated from 138 plants in the Hokkaido area (Taisetsu, Hidaka, Shiretoko, Abashiri and Botanical Garden of Hokkaido University) on high osmotic agar plates. In order to select strains belonging to genus Saccharomyces from the budding yeasts, we constructed two primer pairs specific to Saccharomyces spp. within the D l/D 2 domain of 26 S rRNA gene. Using colony PCR with these specific primers, we obtained 24 tentative strains of Saccharomyces spp. Homology search studies and phylogenetic analysis based on the gene sequences of 26 SrRNA gene D 1/D 2 domain of these tentative strains demonstrated that 23 strains could be identified as Saccharomyces spp. We examined their abilities for ethanol, aroma and CO₂ production, sugar assimilability, and their invertase activities. Distinctive isolates, S. cerevisiae AHU 4438, S. paradoxus AHU 4432, and S. kluyveri AHU 4436, were applied for bread making and the former two strains were evaluated as acceptable regarding bread volume and a distinctive aroma with full body when they were baked after a 40% longer final fermentation time.