The two nightshades Solanum ochranthum and S. juglandifolium show genetic and morphological similarities to the tomatoes (Solanum sect. Lycopersicon), but are isolated from them by strong reproductive barriers. Their genetic relationships to tomato and other Solanum species were investigated using comparative genetic linkage maps obtained from an interspecific F(2) S. ochranthum x S. juglandifolium population. Sixty-six plants were screened using a total of 132 markers--CAPs, RFLPs and SSRs--previously mapped in tomato. Twelve linkage groups were identified, generally corresponding to the expected (syntenic) tomato chromosomes, with two exceptions. Chromosome 1 was composed of two linkage groups and chromosomes 8 and 12 were connected in one large linkage group, indicating a likely reciprocal translocation differentiating the two parental genomes. The total map length comprised 790 cM, representing a 42% reduction in recombination rate relative to the tomato reference map. Transmission ratio distortion affected one-third of the genome, with 13 putative TRD loci identified on 9 out of 12 chromosomes. Most regions were collinear with the tomato reference maps, including the long arm of chromosome 10, which is inverted relative to two other tomato-like nightshades, S. lycopersicoides and S. sitiens. The results support the status of S. ochranthum and S. juglandifolium as the nearest outgroup to the tomatoes and imply they are more closely related to cultivated tomato than predicted from crossing relationships, thus encouraging further attempts at hybridization and introgression between them.