Solanum nigrum (L.) is an important herbaceous medicinal plant belonging to family Solanaceae. It is antiseptic, antibacterial, antidiuretic and anti-inflammatory. In the present study, stem nodal segments of Solanum nigrum were used as explants to induce callus formation on Murashige and Skoog basal medium fortified with 30 g/l sucrose, solidified with 6 g/l agar and supplemented with BA and 2,4-D under biotic (autoclaved Fusarium  oxysporum culture filtrate) and abiotic elicitation (mannitol induced osmotic stress). Results revealed that the callus vigor of growth, color, texture and the nature of the surface of fresh and dry weight, size of callus and contents of some secondary metabolites were affected by the action and interaction and concentration of the two applied elicitors. Elicitation, in general, enhanced the biosynthesis and accumulation of secondary metabolites in callus tissues. Alkaloids were increased from 30 mg/g in dry callus powder (control) to 200 mg/g in calli treated with 1% fungal filtrate and 100 mM mannitol. Flavonoids increased from 20 mg/g in dry callus powder (control) to 410 mg/g  in calli treated with 1% fungal filtrate without mannitol. Tannins increased from 10 mg/g in dry callus powder (control) to 310 mg/g in calli treated with 0.5% fungal filtrate and 50 mM mannitol or 1% fungal filtrate with or without 50 mM mannitol. The obtained results indicated also that elicited calli exhibited in general much more pronounced antibacterial effect against the four tested pathogenic bacterial strains (Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa and Escherichia coli). The obtained results indicate the possibility of maximizing the production of valuable phytopharmaceutics in vitro using biotic and abiotic elicitation.