The species Trifolium polymorphum Poiret is endemic to Uruguay and is widespread in native grasslands throughout the country. Preliminary observations suggested that the aerial flowers are chasmogamous (open at maturity for potential cross-pollination) while the basal flowers are cleistogamous. Several approaches have been practised to determine the reproductive system of forage legumes by the aid of co-dominant markers (Real et al., 2004; Dalla Rizza et al., 2004). The aim of this study is to explore cross-species amplification as a quick approach to obtain co-dominant markers to study the breeding system of T. polymorphum