The turf-type bermudagrasses include diploid Cynodon transvaalensis Burtt Davy, tetraploid C. dactylon (L.) Pers., and sterile triploid hybrids produced by crosses of these species. The objective of thiis study was to develop a set of mierosatellite markers that could distinguish among commercially important turf-type cultivars. A genomic library enriched for tfie [CA/GT]^ repeat motif was constructed from DNA of the Tifway' fiybrid and sequenced to identify mierosatellite regions. Twenty-five microsatellite-flanking primer sets were developed and used to genotype two plant introductions and 12 turf-type cultivars. These primer sets produced an average of 10 amplicons across tfie 14 DNA templates. Sequences of selected amplicons revealed polymorphism resulting from expansion and/or contraction of the microsatellite and from indel mutations in the microsatellite flanking regions. As few as two primer sets were sufficient to differentiate all unrelated introduction lines and cultivars. Tfie primer sets failed to distinguish among closely related cultivars developed by selection of natural variants, but one primer set uniquely distinguished the cultivar TifEagle from its irradiated parent cultivar. Tfiese genomic microsatellites were not derived from gene coding sequences and will supplement tfie existing expressed sequence tag (EST)-based bermudagrass microsatellites. Tfiey will be most useful for evaluating tfie genetic diversity of Cynodon accessions and distinguishing among cultivars that exploit thils diversity.