Development and characterization of microsattelite Loci for the harvested limpets Patella Candei (D'orbigny, 1839) and Patella Aspera (Röding, 1798) using 454 sequencing

World Congress of Malacology, Ponta Delgada, July 22-28, 2013.There is growing consensus that anthropogenic activities are impacting the structure and functioning of marine ecosystems and that these can have profound community level effects, particularly when targeting keystone species. Limpet harvesting in Azores (NE Atlantic) has been taking place probably since the islands were first colonized. These species are highly exploited and the stocks in most islands have declined steadily with catastrophic effects on coastal communities. They are a locally important resource but also ecologically important species and require prioritizing conservation strategies. Such strategies should be su pported by reliable data on the structure and dynamics of their populations, so that ecological hotspots are identified and protected. Characterizing the genetic diversity and structure of marine exploited populations is thus of paramount importance to identify such units of conservation. Here we describe and develop species-specific microsatellite markers for the harvested limpets Patella candei and Patella aspera using whole genome shotgun 454 sequencing. A total of 309 bioinformatic-validated pairs of primers were obtained from P. aspera microsatellite enriched library. The optimization of the amplification conditions of selectect microsatellites (simplex and multiplex reactions) was performed in a gradient thermal cycler to optimize locus-specific amplification conditions and test their utility as genetic markers. Forty pairs of primers were tested, and about 28 revealed to be polymorphic. Using the same procedure, a total of 107 pairs of primers were validated for P. candei of which 15 turned out to be polymorphic. These novel genetic markers can be used to study the population genetic structure and evolutionary history of both patellid species e.g. levels of genetic variability within and between populations, and thus to contribute for stock conservation and management along their distributional area