Ethanol tolerance is crucial for the oenological yeasts.Rosa roxburghiiTratt, a Rosaceae plant native to China, is rich in nutritional and medicinal ingredients. In this study, ethanol-tolerant non-Saccharomycesyeasts were screened, and their oenological properties were further evaluated. Three ethanol-tolerant yeast strains (designated as C6, F112, and F15), which could tolerate 12% (v/v) ethanol treatment, were isolated fromR. roxburghii, and identified asCandida tropicalis,Pichia guilliermondii, andWickerhamomyces anomalus, respectively. The winemaking condition tolerances of these ethanol-tolerant yeast strains were similar to those ofSaccharomyces cerevisiaeX16. However, their growth, sugar metabolic performance and sulphureted hydrogen activities, were different. The β-glucosidase production ability of strainW. anomalusF15 was lower than that ofS. cerevisiaeX16, and strains ofC. tropicalisC6 andP. guilliermondiiF112 were similar toS. cerevisiaeX16. Electronic sensory properties of theR. roxburghiiwines fermented using ethanol-tolerant yeasts together withS. cerevisiaeshowed no significant differences. However, the mixed inoculation of the ethanol-tolerant yeast strains withS. cerevisiaecould regulate the volatile aroma characteristics of the fermentedR. roxburghiiwine, enriching and enhancing the aroma flavor. Therefore, the selected ethanol-tolerant yeasts have the potential for application in the production of uniqueR. roxburghiiwine.