|A 527-marker linkage map of Saccharum spontaneum L. 'SES 208' (2n = 64) was established by analyzing 208 single-dose (SD) arbitrarily primed PCR polymorphisms, 234 single-dose RFLPS, 41 double-dose (DD) and 1 triple-dose (TD) polymorphisms. A map hypothesis constructed using these markers (minimum L.O.D. = 4.00, theta = 0.25 M) had 64 linkage groups with 13 SD, 9 DD, and 1 TD markers unlinked. This is the first complete linkage map (i.e., number of linkage groups observed equals number of linkage groups expected) constructed for any complex polyploid species with no known diploid relatives. Eight chromosome homology groups were identified by using DD and TD fragments as well as SD RFLPs that identified more than one linkage group. Linkages in repulsion phase were absent from the map, as found in two previous genetic studies of this species. Together, these data demonstrate that SES 208 displayed polysomic segregation, a genetic behaviour typical of autopolyploid species. As with previous studies, it was concluded that SES 208 behaved like an auto-octaploid, which was also in agreement with the number of homology groups observed. A chi-square was used to test whether the 527 markers were randomly distributed throughout the genome: both arbitrarily primed PCR markers and RFLPs had a distribution that was statistically indistinguishable from random. The integrated arbitrarily primed PCR-RFLP map had a predicted genomic coverage of 93% (considering only 442 SD polymorphisms) and an average interval between markers of 6 cM. SD markers were used to estimate the genome size of SES 208 at approximately 33 Morgans.