Agrobacteriam rhizogenes-induced medicinal plant hairy root cultures are alternative tools for production of bioactive compounds. Hairy root induction in V. officinalis was studied using various A. rhizogenes strains, A4, ATCC 15834, ATCC 11325 and A13 along with three different induction media, half strength Murashig and skoog (½ MS), Gamborg’s B5 and Linsmaier and Skoog (LS). Transgenic nature of the hairy roots was confirmed by PCR using specific primers for rolB and rolC genes. Different Agrobacterium strains varied in their capability for induction of hairy roots in V. officinalis of which, A13 was not able to induce hairy roots. There was a significant interaction between different induction media and A. rhizogenes strains in terms of transformation efficiency. A4 showed the greatest transformation frequency (54%) in LS medium, while the best medium for ATCC 15834 was ½ MS with 40.67% efficiency. ATCC 11325 showed similar transformation frequency (9.67%) in both ½ MS and LS media, but did not induce any hairy root in B5 medium. on the other hand, respecting hairy roots dry weight, there was no statistically significant difference among different Agrobacterium strains.

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