Using Daqu as a saccharification- fermentation agent is one of the main technological characteristics of traditional Chinese liquor. The microbial components in Daqu were rich, including various types of mold, yeast and bacteria. These complex microbial systems provided necessary microorganisms, enzymes and flavor substances for the fermentation of liquor. As an essential microorganism in all alcoholic fermentation, yeast also played an important role in the production of Daqu. To explore the functional microorganism in Daqu, the yeasts, isolated from Daqu in different periods , were compared and analyzed by 26S rRNA D1 / D2 region sequences, and 260 yeasts were isolated and identified, belonging to 22 species. The yeasts were mainly of Wickerhamomyces anomalus, Candida orthopsilosis, Meyerozyma guilliermondii, Pichia caribbica, Saccharomycopsis fibμLigera, Cryptococcus neoformans var. Grubii, Clavispora lusitaniae, Saccharomyces cerevisiae. Single strand conformation polymorphism (SSCP) analysis was performed on the results of sequence alignmentTheirinterspecies differences were determined and the M-L system tree was also constructed. In addition, the composition and variation of yeasts in different periods of Daqu production were preliminarily investigated according to the results of isolation and identification. These results provided a basis for further research on brewing microorganisms of traditional Chinese liquor.