Creating Microsatellite Primers for \u3cem\u3eBotrychium\u3c/em\u3e

Botrychium sensu stricto (Ophioglossaceae) is a genus with cryptic species that are difficult to delineate on the basis of their limited morphology and high incidence of hybridization events. Various means have been used to determine the phylogeny of Botrychium based on both genetic and morphological factors. We designed species-specific microsatellite primers to better determine species relationships between the allotetraploid Botrychium matricariifolium and the diploid species Botrychium angustisegmentum. We examined seventeen individuals, from both B. matricariifolium and B. angustisegmentum. We found that both B. matricariifolium and B. angustisegmentum amplified on the majority of DNA on our gels for eight of twelve runs. Amplified loci on the Beckman Coulter CEQ 8000 runs were associated with bands visualized on the gel. Preliminary results with a high percentage of missing data showed a genetic difference between the two species. There was little variation within the populations, however we may see more by using our primers to test other populations of B. matricariifolium and B. angustisegmentum may also prove beneficial to look at other species entirely, particularly Botrychium pallidum