Abstract:A total of five hundred and forty-nine isolates of endophytic fungi isolated from Taxus mairei were screened for the bioactivity targeted against Pythium ultimum and Rhizoctonia solani. Of 549 isolates, 20 isolates (about 3.6% of the total isolates) showed significant activity against P.ultimum and 15 isolates (about 2.8% of the total isolates) against R.solani with ID50 beyond 10. The isolate F0028, which has the highest inhibitory rate against P.ultimum (ID50 of 278±15.0) and R.solani (ID50 of 108±18.6) was identified as Aspergillus clavatonanicus. Its crude fermentation broths also showed strong inhibitory activity at pH value range from 1.0 to 7.0 and retained certain inhibitory activity of (65±3.8)% when autoclaved. Five compounds were isolated from the strain F0028 crude fermentation broths by HPLC. The results of bioassays revealed that all the five compounds have certain inhibitory activity with EC50 between 7.0 and 45 μg/mL against two pathogenic fungi.