Optimizing the composition of inorganic nutrients of the media for hairy root cultures is essential to gain high production of secondary metabolites. It has been reported that concentration of nitrogen and appropriate ratio of nitrogen sources in the culture medium greatly affected the growth and production of secondary metabolites in hairy root cultures. In the present study, the effect of different basal media and various NH4+ to NO3- ratios in the culture medium on the growth of hairy root cultures of V. officinalis are discussed.