Culture conditions have been established for the induction of callus from different explants of Paspalum simplex. Fast-growing calli were obtained from hypocotyls and roots excised from 5-day-old seedlings on culture medium containing 2,4-dichlorophenoxyacetic acid and kinetin. Rapid plant regeneration from both apomictic and sexual lines was achieved when the medium was supplemented with alfa-naphthaleneacetic acid and benzyladenine. Restriction fragment length polymorphism analysis of the apomixis-controlling region of the regenerated plants showed an absence of restriction site variation for the loci analysed, whereas various degrees of variation were detected for the DNA methylation sites of the same loci