Research into plant-derived endophytic fungi had grown in recent decades. Endophytic fungi still had enormous potential to inspire and influence modern agriculture. The aim of the present study was to screen for antimicrobial activity of endophytic fungi isolated from surface sterilized leaves, stems and fruits of Eucommia ulmoides oliv. to Gibberella zeae. The fermentation broths from 32 isolated fungi were tested for antimicrobial activity by the mycelium growth rate method. The results showed that fermentation broths from the majority of isolates exhibited antifungal activity and the strongest antimicrobial activity was exhibited by strain DZGS01 against Gibberella zeae, showing 66.47 % inhibition at 1 mg/mL , compared to 62.36 % for 12 µg/mL carbendazim. Moreover, compared to water control , the control efficacy of the fermentation broth of the DZGS01 was 63.16 % by pot experiment. And the endophytic fungus DZGS01 was belonged to Myrothecium based on rDNA sequencing of ITS region .The results indicate that the endophytic fungus DZGS01 from the plant Eucommia ulmoides oliv. was a promising source of novel bioactive inhibitors against Gibberella zeae.