The aim of the study was to test the synergistic activity of zearalenone and thidiazuron with commonly used regulators (6-benzylaminopurine, 2,4-dichlorophenoxyacetic acid) in callus tissue growth and the regeneration processes of winter oilseed rape and winter wheat. Oilseed rape callus was obtained from the upper fragments of seedling hypocotyls. Wheat callus tissue was obtained from immature embryos. Zearalenone, in combination with 6-benzylaminopurine, stimulated the regeneration of oilseed rape and, in combination with 2,4-dichlorophenoxyacetic acid, the regeneration of wheat. An additional application of thidiazuron to these media increased the percentage of shoot regeneration from callus in both species tested. Zearalenone is proposed as a new regulator in in vitro plant cultures for use in protocols of plant regeneration.