Abstract: Solanesol is an important material for synthesizing ubiquinone. An efficient transformation system was established in Nicotiana tabacum by use of Agrobacterium rhizogenes W.T15834.The content of solanesol was detected. Hairy roots could grow normally on solid or liquid MS medium without a growth regulator. However, the hairy roots grew rapidly and no callus formed during subculture in liquid medium. The transformation of hairy roots was confirmed by PCR and mannopine assay. By use of improved HPLC, we obtained a content of solanesol of almost 43.2% in hairy roots that was 1.12 times that in roots of tobacco seedlings, but still lower than that in discarded tobacco leaves; the content of solanesol is normally higher in liquid culture than in solid culture. This study aims to provide a basis for future research in taking advantage of hairy root technology to produce solanesol.