Moso bamboo (Phyllostachys pubescens, Gramineae) is a well-known medicinal and edible plant found in China with various bioactivities, but few systematic studies address the utilization of its anti-fungal activity. The extract of moso bamboo leaf showed good anti-fungal activity to Phytophthora capsici, Fusarium graminearum, Valsa mali Miyabe et Yamada, Botryosphaeria dothidea, Venturia nashicola, and Botrytis cinerea Pers, with inhibitory rate of 100.00%, 75.12%, 60.66%, 57.24%, 44.62%, and 30.16%, respectively. Anti-fungal activity was different by the difference of samples picking time and location. The extract showed good synergistic effects with carbendazim at the ratios of 9:1 and 15:1 (extract : carbendazim), and the co-toxicity coefficients were 124.4 and 139.95. Compound 2 was isolated and identified as the main active component, with the EC50 value of 11.02 mg L-1. Then, the extract was formulated as a 10% emulsion in water, which was stable and had no acute toxic effects. Moreover, a field trial about this formulation was assayed to control pepper phytophthora blight, with the control effect of 85.60%. These data provided a better understanding of the anti-fungal activity and relevant active component of moso bamboo leaf extract. Taken together, our findings illustrated that bamboo leaf extract could be developed and utilized as a botanical fungicide or fungicide adjuvant.