This study was aimed to explore the resource diversity and microbial inhibition activity of endophytic fungi from medicinal plant Plantago asiatica L. The endophytic fungi were isolated from the root, stem and leaf of the host by tissue inoculation culture and five plant pathogenic fungi and four bacteria strains used as indicating microbes to test microbial inhibition activity by agar plate antagonistic action and modified agar gel diffusion methods. The results indicated that thirteen fungal endophytic strains were isolated from the host. Most of them came from stem, then leaf, and root as the least in number. The isolated strains attribute to five genera, two fam-ilies, and two orders based on morphological characteristics. For the isolated strains, eleven of them were found to have some microbial inhibition activities against one or more indicating fungi, making up 84.6% of the total iso-lates. Six isolated strains had some antimicrobial activities against one or more indicating bacteria, amounting to 46.2% of the total isolates. Three isolated active strains, which are PAEFS001, PAEFS007 and PAEFS008, ex-hibited evident inhibition activities against five kinds of pathogenic fungi used in the trials respectively. The strain of PAEFS001 ascribed to Ozonium sp. Both strains of PAEFS007 and PAEFS008 ascribed to Aspergillus sp. One active strain of PAEFS003 showed evident antibacterial activities to Bacillus subtilis and Staphyloccus aureus, which belonged to Fusidium sp. The endophytic fungi from medicinal plant Plantago asiatica L. have evident an-timicrobial activities. Their inhibition activities against pathogenic fungi have relatively broad spectrum. And their inhibition activities to both Bacillus subtilis and Staphyloccus aureus as G+ are evident and have certain selectivi-ty. It is feasible to find new bioactive compounds associated with endophytic fungi from Plantago asiatica L. Fur-ther research and development of the endophyic fungi will be important for the integrated utilization of the host.