Osmophilic yeast in honey at home and abroad was detected, and the strains isolated from some samples were identified and their characteristics tested. Methods A total of 916 batches of domestic and imported honey were selected, and osmophilic yeast was counted according to GB 14963-2011 "National Standard Honey for Food Safety" and GB 4789.2-2022 "Determination of the total number of colonies for microbial Inspection of National Standard Food Safety", and the detection rate of osmophilic yeast was calculated. The isolated strains were amplified by 26S rDNA PCR, and the sequences were sequenced and compared to determine the species. The ability of some yeast strains to tolerate high sugar and produce alcohol was tested by absorbance and potassium bichromate colorimetry. Results In 916 batches of honey, 36 batches of osmophilic yeast were detected, and the detection rate was 3.93%. Among 786 batches of domestic honey, 33 batches of osmophilic yeast were detected, the detection rate was 4.20%, and the osmophilic yeast count was 2.5×102 ~ 7×104 CFU/g. Three osmophilic yeasts were detected in 130 batches of imported honey, the detection rate was 2.31%, and the osmophilic yeast count was 1×101 ~ 7×101 CFU/g. Five osmophilic yeast strains were isolated, one was Moesziomyces aphidis/Pseudozyma sp., and three were Zygosaccharomyces mellis. One was Candida magnoliae , and all of the tested strains were well tolerant to high glucose concentrations and had some capacity to produce alcohol. Conclusion Honey may be contaminated by osmophilic yeast, so it is necessary to strengthen its control and management. Honey can be used as a source for screening osmolal-resistant strains, and the selection of osmophilic yeast is helpful to the development of honey food fermentation industry and the research and development of excellent products.