This study was proposed to investigate the in-vitro antifungal, antibacterial, insecticidal, cytotoxic and phytotoxic potential of ethanolic extract of twigs of Taxus wallichiana Zucc. Traditionally, this plant is used in folk medicines in Pakistan, for the treatment of various microbial infections. The crude sub fractions of ethanolic extracts of the plant were tested against six bacterial and six fungal strains using hole diffusion and macro-dilution methods, respectively. The n-hexane and ethyl acetate fractions showed 15 and 24 mm zones of inhibition against Staphylococcus aureus, while the chloroform fraction exhibited 35 mm inhibition against Shigella flexenari. In case of antifungal activity, both the chloroform and methanol fractions showed minimum inhibitory concentration at the concentration level of 15 μg/mL against Microsporum canis, while ethyl acetate fraction exhibited antifungal activity against Candida albicans and Candida glaberata at 27 and 13 μg/mL concentrations, respectively. The rest of the fractions showed no antifungal activity. The cytotoxicity assay revealed that all the fractions have activity against the shrimp. The n-hexane, chloroform and methanol fractions were found active against Rhyzopertha dominica, Callos bruchuanalis and Tribolium castaneum. In case of phytotoxicity assay, all fractions showed significant activity at the concentration of 500 μg/ml.