To establish the hairy roots culture system of Patrinia scabioscaefolia Fisch and analyze the flavone content in hairy roots culturing system and its antibacterial role of basic research.Methods:Hairy root of Patrinia scabioscaefolia Fisch were induced by the infection of four Agrobacterium rhizogenes strains: R15834, R1601, R1000, and A4. The infecting time, pre-culture and co-culture time on growth of hairy root were studied, and the hairy roots were detected by PCR. The purity of flavone was determined by UV, and determination of inhibition was tested by disk diffusion method.Results:The highest induction frequency was obtained from stem segment with 48 h pre-culture and co-culture which were induced by A4 for 8 min. The transformation frequency and proliferation rate could be raised by culture media with sucrose concentration of 40 g/L. The PCR examination result showed that rolB genes were successfully inserted into the hairy roots of Patrinia scabioscaefolia Fisch.Conclusion:Hairy root of flavonoids content is higher than that of the natural content and that the aqueous extrats had obvious antibacterial effect against Staphylococcus aureus.