Abstract: Objective: To avoid browning and spoilage caused by microbial growth, there is an urgent need to carry out researches related to the spoilage species of Stropharia rugosoannulata, and to provide a theoretical basis for exploring storage and preservation techniques. Methods: The spoilage fungi of S. rugosoannulata were isolated and purified, and the dominant spoilage fungi were selected by corrosivity analysis and identified by morphology and molecular biology methods. Results: The spoilage fungi from S. rugosoannulata were carried out to 16 strains of Fusarium, 14 strains of Aspergillus and 4 strains of Rhizopus among them. Four decay-causing fungi were selected to have a rot-causing effect, and identified by morphology and molecular biology as Fusarium pseudoanthophilum, Aspergillus niger, Rhizopus azygosporus and Talaromyces purpureogenus, respectively. The growth characteristics showed that the optimal temperature was 25~28 ℃. The spoilage fungi grew slowly with the temperature of 4~10 ℃ and 35~60 ℃, and the lethal temperature was 60 ℃. The optimal pH was about 7.0 and the spoilage fungi could not grow when the pH was 13.0. Conclusion: Post-harvest storage at low temperature and suitable alkaline environment could inhibit the growth and reproduction of spoilage fungi, which was conducive to the storage and preservation of S. rugosoannulata.