The Dilatata group of Paspalum is a polyploid complex native to the grasslands of temperate South America. A pentaploid apomictic biotype of P. dilatatum is a widely recognized forage grass; however, the complex includes apomictic tetra- and hexaploids along with sexual allotetraploids (P. urvillei, P. dasypleurum, P. dilatatum ssp. flavescens, and biotypes Virasoro and Vacaria of P. dilatatum) which are suspected to be hybrids between different combinations of biotypes. Fifteen primer pairs for nuclear microsatellite loci were developed from a (CA)n-enriched genomic library. A single, low stutter, robust two-step PCR profile was used for all the primer pairs. Twelve primer pairs were analysed for their variability and transferability among all the sexual biotypes of the Dilatata group and the common apomictic pentaploid. Six primer pairs amplified more than one locus, and sequence and segregation evidence suggest that the additional bands correspond to homeologous loci. No close linkage was found among the 16 loci amplified in the tetraploids. One variable chloroplast microsatellite is also reported. All loci were successfully amplified from most members of the Dilatata group and variability was recorded for all the biotypes analysed. The set of loci reported here provide highly variable markers for intra-biotypic population studies, biotype-specific markers for the analysis of hybrid apomicts and a biotype-specific chloroplast marker.