In the present study, Fenugreek (Trigonella foenum graecum) seeds were surface sterilized and allowed to germinate on hormone free Murashige and Skoog's culture media for one week. Hypocotyls were cut into 0.5 mm length segments.  Callus morphogenesis was achieved from the hypocotyls segments Using Murashige and Skoog's culture media supplemented with benzyl adenine and naphthalene acetic acid. The best callus biomass was obtained with 2 mg/l (BA) plus 1.5 mg/L (NAA) and the growth biomass was measured after 4 weeks. This concentration was then used to plot the growth curve of callus under both dark and light growth conditions. Calli obtained under light conditions exhibited a greenish yellow pigmentation, generally accumulated better biomass and their cells contained numerous and well formed discoid plastids. Calli obtained after 3, 6 and 9 weeks of growth were harvested, dried, and grinded into a fine powder and then extracted with ethanol 80%. Antibacterial activity of various calli extracts was checked by the disc-plate diffusion method and the results obtained have indicated that both hypocotyls and calli obtained from these explants exhibited activity against the tested bacteria and it was generally observed that calli were more potent than hypocotyls and that calli induced under light conditions were more potent in some cases than dark-induced calli in inhibiting the bacterial growth.