Berberis orthobotrys Bienerat ex C.K. Schneider (BOB) is broadly utilized as a medicinal plant in Asia. In present work, various solvents (water, methanol, ethanol) were used as solvent in the extraction of BOB. Then, total phenol, flavonoid and tannin contents of dried BOB were investigated. Additionally, in vitro and antioxidant capacities and antibacterial and antifungal activities of different solvent extracts were also evaluated. The extract obtained by 80% methanol had the maximum amount of total phenolic, flavonoid and tannin contents, while, the minimum amount of these compounds were obtained for aqueous extract of BOB. It was found that the anti-DPPH radical activity and reducing power of methanolic extract were higher than other samples. The effect of BOB extract on the oxidation of sunflower oil demonstrated that BOB can inhibit the oxidative reactions. The evaluation of antibacterial and antifungal activities of different BOB extracts exhibited that methanolic extract had significant inhibition activity against tested bacteria and fungus. In conclusion, BOB can be used as a potential source of natural antioxidant and antibacterial agent for food and pharmaceutical systems.