Technical Abstract: The increasing use of secondary water sources for irrigation, which is highly linked to salinization, creates a demand for salt tolerant turf. Seashore paspalum (Paspalum vaginatum Swartz) is a warm-season turfgrass that survives in sand dunes along coastal sites, brackish ponds, and in estuaries. Some seashore paspalum accessions/cultivars are far more salt tolerant than others. In order to identify genetic regions that are associated with salt tolerance, molecular tools must be developed. In this study, genomic libraries, enriched for microsatellites, were generated using the salt tolerant accession HI33. High throughput sequencing and subsequent assembling of these libraries resulted in 18,967 contigs and 158,595 singletons. The number of simple sequence repeats (SSR) detected in contigs and in singletons was 3,511 and 31,949, respectively, and the number of primer sets designed within each group was 937 and 1,667. A total of 80 SSR markers, including five markers previously developed, were used to assess genetic relationships among 18 Paspalum accessions. Two major clusters were identified from the seashore paspalum accessions. Accessions that are likely polyploids, all with coarse leaves, grouped together whereas accessions with fine-to mid-fine leaves formed a second group. Some cultivars, such as Seadwarf, Temple 2, SeaIsle 1, and Taliaferra reported to have been collected from all over the world, were found to be very genetically similar. Furthermore, 33 seashore paspalum SSR markers cross-amplify in bahiagrass (Paspalum notatum Flugge) and these markers can be a useful tool in this species.