The yielding and yield quality are affected by interactions of complex dependencies between the plant, the habitat, and the agricultural practice. The goals of this study were to determine grain yield, grain quality, and winter wheat root infestation by Gaeumannomyces graminis var. tritici in crop rotation and monoculture. The experiment was established with the method of equiva- lent subblocks (25 m × 6 m) in three replications. The main experimental factor was the crop- ping systems (CS), and the second-order experimental factor was the tillage systems (TS). Winter wheat was sown in the following crop rotations: (i) CR-A: potato – winter wheat – peas – winter barley; (ii) CR-B: peas – winter barley – winter wheat – spring wheat; and (iii) MON: multi-year wheat monoculture. The following tillage systems were applied in CR-A, CR-B, and MON: (a) conventional tillage (CT); (b) reduced tillage (RT); and (c) no-tillage (NT). Wheat grain yield was affected by CS more than by TS; its highest value was recorded in CR-A followed by CR-B, whereas the lowest one – in MON. Higher grain yield was also obtained in NT and RT than in CT. The CR-A and CR-B as well as CT systems were found to elicit posi- tive effects on the gluten content of wheat grain, grain weight per volume, Zeleny’s sedimenta- tion index, grain uniformity, and ash content of the grain. Wheat root infestation by Gaeuman- nomyces graminis var. tritici was more severe in MON than in CR-A and CR-B, and in NT than in CT and RT.