The production of taxol endophytic fungi was improved through optimizing the fermentation conditions of WY2, which would support the foundation for following study. The culture medium, the initial pH, fermentation temperature, shaker speed and the incubation time have been explored, and detected the influence on the Taxus chinensis taxol production by endophytic fungi. The results show that YES medium was the best culture, and the initial pH 6.0, cultivate temperature 30 °C, shaker rate 200 r/min was the best fermentation conditions. The effects of some precursor on production of taxol in endophytic fungi strain WY2 also were studied. By the orthogonal experiment, it showed that when add 20 mg/l phenylalanine, 10 mg/l of ammonium acetate, 5 mg/l tyrosine, 20 mg/l of glycine to medium, taxol production (547 μg/l) was increased by 337%, compared to that of initial production The optimized culture condition and the appropriate precursor can be used to improve the production of taxol, meeting the demand of medical application.