Abstract: To assess the diversity of fungi in soil of Great Khingan forest, the forest surface soil around the Ecosystem Station ofGreat Khingan was collected. Then based on ITS gene,the Miseq high-throughput sequencing technique was applied to discoverthe diversity of fungi in the soil. The resuhs showed that obtained 3 260 ITS sequences could be usable. At the divergence levelof 0.03, these sequences were divided into 51 OTU. The alpha diversity indexes namely Shannon,coverage, Chao value and acevalue were 0.8189,85.60% ,85 and 155, respectively. The ITS sequences were originated from Basidiomycota, Ascomycota,Zygomycota and some unknown tungal taxa, of which the predominated was Basidiomycota with the highest ratio of 97.91%.