In this study, we isolated an endophytic Streptomyces sp. strain, WP-1, from surface-sterilized barks of Pinus dabeshanensis, an endangered Chinese plant. WP-1 showed strong antifungal activity against diverse pathogenic fungi, such as Fusarium oxysporum, Rhizoctonia solani, Phytophthora infestan, and Candida albicans. Based on phylogenetic analyses, preliminary identification suggested that the WP-1 strain belonged to the genus Streptomyces. Column chromatogram and HPLC were employed to isolate the primary antifungal component from the culture medium of WP-1, and it was identified as the methylpentaene macrolide antibiotic, fungichromin (FC). In this study, for the first time, using in vitro bioassay studies, we revealed that FC strongly inhibited mycelial growth and conidia germination in Fusarium oxysporum. The median inhibitory concentration of FC was found to be 3.80 mg/L. The fermentation conditions of the WP-1 strain were further investigated to improve FC production. We found that supplementation of the synthetic medium with oils (soybean oil, oleic acid, and so on), particularly during the initial stage of fermentation, significantly increased the FC yield. Ammonium-trapping agent (magnesium phosphate) was used as an additive to increase FC yield to 5741.7 mg/L. It was 2.9-fold more as compared to the highest FC yield reported so far where Streptomyces padanus PMS-702 was used for FC production.