Production of flavonoids in madder callus culture (Rubia tinctorum L.) was dependent on culture conditions and culture media composition. The content of flavonoids increased in calli maintained on media supplemented with NAA (4 mg.l-1) or NAA:BAP (4 mg.l-1 a 1 mg.l-1) in 16 h photoperiod. Flavonoids represented 2.08 – 2.25 % of callus dry mass. The presence of Cd(NO3)2 (3.1 or 31.0 mg.l-1 concentrations) negatively influenced callus growth, but enhaced the percentage of dry mass in callus cells. During 42 days of culture an increase of cadmium accumulation and even of flavonoids has been observed. The most considerable influence of CdCl2 or Cd(NO3)2 on flavonoids content has been shown in short-term experiments after 48 h of callus culture. More distinct influence has been observed under the treatment with CdCl2 (0.005 mg.l-1) in comparison with Cd(NO3)2.