The objectives of these experiments were to evaluate male sterility (MS) and 2n pollen frequency in 77 4x×2x families and 26 4x×4x families. The 2x parents were haploid-species hybrids and the 4x parents mainly clones with S. tuberosum ssp. tuberosum (tbr) and S. tuberosum ssp. andigena (adg) in their genetic background. The female parents had different cytoplasms: adg, tbr, S. demissum (dms), and S. stoloniferum (sto). Families from female parents with adg or dms cytoplasm did not have MS plants. However, families derived from crosses of many tbr females with 2x hybrids and a 4x adg clone had a high percentage of MS plants; all the progenies from cv. Serrana (sto cytoplasm) possessed tetrad sterility. These results can be explained as due to an interaction between a dominant male sterility (Ms) gene with tbr and sto cytoplasms. Some MS 4x hybrids (tbr×adg) used as females had the Ms gene, but male fertility was restored in some plants of their progenies when they were crossed with 4x tbr clones. This indicates that some tbr clones have a fertility restorer gene (Rt). The results from both 4x×2x and 4x×4x families fit the expected ratios assuming chromatid segregation for both the Ms and Rt loci. The gene frequency of parallel spindles was estimated in the 4x population as 0.74. © 1993 Kluwer Academic Publishers.