RESUMO
PREMISE OF THE STUDY: Microsatellite markers were developed for Polypodium appalachianum (Polypodiaceae) to facilitate investigation of species boundaries between P. appalachianum and its putative hybrid, P. virginianum, and potentially among other members of the Miocene-age P. vulgare species complex. METHODS AND RESULTS: Forty-eight primer pairs were designed from Illumina data and screened for successful amplification. Sixteen pairs were genotyped and evaluated for variability within and among three populations in North Carolina, Vermont, and New Hampshire. Twelve of these primer pairs were reliable and polymorphic, exhibiting one to 10 alleles per locus. Cross-species amplification experiments were conducted for P. virginianum and four additional close relatives from the P. vulgare complex in order to maximize information about likely utility within a phylogenetic context. CONCLUSIONS: These microsatellite markers will be useful in population genetics and species boundaries studies of P. appalachianum and P. virginianum, and likely in other species within the P. vulgare complex.