The effect of UV radiation (254 and 366 nm) on isoflavonoid production in Genista tinctoria callus cultures was investigated. The production of the separated isoflavonoids in callus culture of G. inctoria was changed in the dependence on wavelength of UV treatment. Genistin, genistein, daidzein, biochanin A in higher levels were formed after UV irradiation in callus culture in comparison with untreated callus culture. Maximal genistin content (3.03%) was reached after treatment of UV 254 nm for 300 s (sampled after 48 h). High genistin content (2.06%) was observed also after 120 s of UV 366 nm radiation (sampled after 48 h). Formononetin was detected only after UV treatment in G. tinctoria callus culture.