Cysteine, gamma-glutamylcysteine, and glutathione and the extractable activity of the enzymes of glutathione biosynthesis, gamma-glutamylcysteine synthetase (EC 6.3.2.2) and glutathione synthetase (EC 6.3.2.3), were measured in roots and leaves of maize seedlings (Zea mays L. cv LG 9) exposed to CdCl2 concentrations up to 200 micromolar. At 50 micromolar Cd2+, gamma-glutamylcysteine contents increased continuously during 4 days up to 21-fold and eightfold of the control in roots and leaves, respectively. Even at 0.5 micromolar Cd2+, the concentration of gamma-glutamylcysteine in the roots was significantly higher than in the control. At 5 micromolar and higher Cd2+ Concentrations, a significant increase in gamma-glutamylcysteine synthetase activity was measured in the roots, whereas in the leaves this enzyme activity was enhanced only at 200 micromolar Cd2+. Labeling of isolated roots with [S-35]sulfate showed that both sulfate assimilation and glutathione synthesis were increased by Cd. The accumulation of gamma-glutamylcysteine in the roots did not affect the root exudation rate of this compound. Our results indicate that maize roots are at least in part autonomous in providing the additional thiols required for phytochelatin synthesis induced by Cd.