Objective To systematically analyze and compare the diversity of endophytic fungi from wild and cultivated Dendrobium officinale, and screen fungal strains with anti-anthracnose activities.Method Endophytic fungi of D. officinale were isolated by tissue block separation method, and they were identified by slide culture method and molecular identification method based on internal transcribed spacer regions of ribosomal DNA (rDNA-ITS). Screening for endophytic fungi against anthracnose was carried out by plate method.Result Totally 236 endophytic fungal strains were isolated from D. officinale(132 from wild plants and 104 from artificially cultivated plants). Total colonization rate (50.46%)，total isolation rate (61.11%) and diversity index (2.95%) of endophytic fungi in wild plants were significantly higher than those in artificially cultivated plants (the corresponding values were 39.35%, 48.14% and 2.49, respectively). We screened out 34 strains of endophytic fungi with inhibitory activities against Colletotrichum gloeosporioides. Amomg them, the strains Ceriporia sp. F102, Rigidoporus sp. F27, Termitomyces sp. F103 and Peniophora sp. F122 showed higher inhibitory effects, and their inhibition rates were 93.06%, 88.20%, 87.48% and 84.57%, respectively.Conclusion The biodiversity and community structure of endophytic fungi of wild D. officinale and artificially cultivated D. officinal had significant differences, and abundant strains with anti-anthracnose activities were obtained from them, which lays the foundation for improving the quality of medicinal materials of artificially cultivated D. officinal and the biocontrol of D. officinale anthracnose.