Hairy root induction in Plantago lanceolata was optimized to take advantage of transformed root cultures. The highest frequency of transformation was achieved using leaf explant, A4 strain, pre-cultivation of explant, 150 µM Acetosyringone, 5 min inoculation, half-strength Murashige and Skoog basal medium as co-cultivation, and half-strength Gamborg's basal medium as a selective medium with 3% sucrose. Among the studied compound encompassing gallic acid, catalpol and apigenin, only the production of gallic acid in hairy roots was affected by 20 mg L^-1 AgNO₃ and 100 mg L^-1 chitosan at 24 hr which yielded 7.63, 4.76-fold increase in its content, respectively. The methanolic extracts of hairy roots elicited by 20 mg L^-1 AgNO₃ exhibited anti-bacterial activity (MIC and MBC = 25 mg mL^-1) against Klebsiella pneumoniae, Proteus vulgaris and Salmonella typhi and anti-bacterial potential of non-elicited hairy roots of P. lanceolata (MIC = 25 mg mL^-1 and MBC = 35 mg mL^-1) were more active against Klebsiella pneumoniae and P. vulgaris than other bacteria. The methanolic extracts of the P. lanceolata hairy roots demonstrated significant cytotoxic activity on colorectal carcinoma cell line (SW-480) with IC₅₀ = 250.65 ± 6.8 µg mL^-1 in comparison to human embryonic kidney (HEK-293) with IC₅₀ = 5263.65 ± 4.6 µg mL^-1. Plantago lanceolata hairy roots showed important biological activity explaining its role in traditional medicine.

Keywords: Plantago lanceolata; Anti-bacterial; anti-cancer activities; elicitors; transgenic roots.