Abstract: In order to analyze the correlation between fungus and physicochemical characteristics of medium-temperature Daqu in Xiangyang, Hubei Province, high-throughput sequencing and pure culture techniques were used to analyze the samples from Hubei Longzhong Wine Industry, and physicochemical and sensory indexes were evaluated, then the influence of fungus on its quality was revealed. The result showed that all the physicochemical indexes of Daqu samples met the requirements of QB/T 4259-2011 "Fragrant Daqu", and there were great differences in esterification and saccharification among samples (P<0.05). Thermomyces, Rhizopus, Dipodascus, Aspergillus Pichia, Hyphopichia and Penicillium were the main fungal genus. The isolated 25 strains were identified as Saccharomycopsis fibuligera and Wickerhamomyces anomalus. The result of correlation showed that Hypericum had a positive effect on the formation of aromatic compounds (P<0.05). Thus, the medium-temperature Daqu in Hubei Longzhong Wine Industry contains a variety of fungi that need to be explored, and the metabolism of some fungi are conducive to the formation of organic sulfides and aromatic compounds.