As an insufficiently utilized energy resource,oil shale is conducive to the formation of characteristic microbial communities due to its special geological origins.However,little is known about fungal diversity in oil shale.Polymerase chain reaction cloning was used to construct the fungal ribosomal deoxyribonucleic acid internal transcribed spacer(r DNA ITS)clone libraries of Huadian Mine in Jilin Province,Maoming Mine in Guangdong Province,and Fushun Mine in Liaoning Province.Pure culture and molecular identification were applied for the isolation of cultivable fungi in fresh oil shale of each mine.Results of clone libraries indicated that each mine had over 50% Ascomycota(58.4%–98.9%)and 1.1%–13.5%unidentified fungi.Fushun Mine and Huadian Mine had 5.9% and 28.1% Basidiomycota,respectively.Huadian Mine showed the highest fungal diversity,followed by Fushun Mine and Maoming Mine.Jaccard indexes showed that the similarities between any two of three fungal communities at the genus level were very low,indicating that fungi in each mine developed independently during the long geological adaptation and formed a community composition fitting the environment.In the fresh oil-shale samples of the three mines,cultivable fungal phyla were consistent with the results of clone libraries.Fifteen genera and several unidentified fungi were identified as Ascomycota and Basidiomycota using pure culture.Penicillium was the only genus found in all three mines.These findings contributed to gaining a clear understanding of current fungal resources in major oil-shale mines in China and provided useful information for relevant studies on isolation of indigenous fungi carrying functional genes from oil shale. As an insufficiently utilized energy resource, oil shale is conducive to the formation of characteristic microbial communities due to its special geological origins. However, little is known about fungal diversity in oil shale. Polymerase chain reaction cloning was used to construct the fungal ribosomal deoxyribonucleic acid internal transcribed spacer (rDNA ITS) clone libraries of Huadian Mine in Jilin Province, Maoming Mine in Guangdong Province, and Fushun Mine in Liaoning Province. Pure culture and molecular identification were applied for the isolation of cultivable fungi in fresh oil shale of each mine .Results of clone libraries indicated that each mine had over 50% Ascomycota (58.4% -98.9%) and 1.1% -13.5% unidentified fungi. Fushun Mine and Huadian Mine had 5.9% and 28.1% Basidiomycota, respectively. fungal diversity, followed by Fushun Mine and Maoming Mine. Jaccard indexes showed that the similarities between any two of three fungal communities at the genus level were very low, indicating that fungi in each mine developedpendent during the long geological adaptation and formed a community composition fitting the environment. the fresh oil-shale samples of the three mines, cultivable fungal phyla were consistent with the results of clone libraries .Fifteen genera and several unidentified fungi were identified as Ascomycota and Basidiomycota using pure culture. Penicillium was the only genus found in all three mines. The findings made to gaining a clear understanding of current fungal resources in major oil-shale mines in China and provided useful information for relevant studies on isolation of indigenous fungi carrying functional genes from oil shale.