The antimicrobial potential of the aqueous and ethanolic extracts of seeds, leaves, roots, callus and in vitro regenerated plantlets leaves of Anethum graveolens have been evaluated against important bacterial strains, Escherichia coli HB101 (MTCC-82), Bacillus subtilis (MTCC-441), Bacillus cereus (MTCC-1306), Micrococcus luteus (MTCC-2452). The antimicrobial activity was determined in aqueous and ethanol extracts using both agar disc diffusion and agar well diffusion method. The ethanolic extracts were found to be more potent than aqueous extracts of all parts of plant studied. The ethanolic extracts of seeds showed strong activity against all bacterial strains. In comparison to in vivo, in vitro plant extracts depicted reduced activity. The phytochemical screening of the plant parts showed that leaves, stems, roots, in vitro callus and regenerated leaves were rich in tannins, terpenoids, cardiac glycosides and flavonoids. Though, the seeds of Anethum have been used traditionally as decoctions or infusions prepared in water to treat various ailments, due to the presence of active component, hence, they show maximum activity. During the present study, we have tested effectiveness of in vivo plant parts as well as in vitro grown callus and leaves, on selected strains of bacteria. These phytochemicals and secondary metabolites could be responsible for the antibacterial activities exhibited by the extract and hence, justify the medicinal uses of A. graveolens by common folks.