Identification and amplification of microsatellite loci in deep-sea tubeworms of the genus Escarpia (Polychaeta, Sibogli
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TECHNICAL NOTE
Identification and amplification of microsatellite loci in deep-sea tubeworms of the genus Escarpia (Polychaeta, Siboglinidae) Dominique A. Cowart • Chunya Huang Stephen W. Schaeffer
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Received: 6 November 2012 / Accepted: 24 November 2012 / Published online: 4 December 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract Deep-sea cold seeps support diverse communities that include vestimentiferan tubeworms of the genus Escarpia. To investigate population structure to understand how deep-sea communities are established, several polymorphic microsatellites were developed using 454 pyrosequencing. Eleven and sixteen loci were identified in Escarpia laminata (n = 129) and Escarpia southwardae (n = 80). Additionally, the amplification of microsatellites is expected to be successful in closely related species and can help resolve their phylogenetic relationships. Nine of the E. laminata loci were amplified across three Escarpia species (n = 229). Four of the twenty-eight loci deviated from Hardy–Weinburg Equilibrium and no linkage disequilibrium was found in the loci tested here. Keywords Vestimentiferan tubeworm Hydrocarbon seep High-throughput marker identification Microsatellite Siboglinid
Cold hydrocarbon seeps fuel specialized deep-sea ecosystems and are home to vestimentiferan tubeworms (Family: Siboglinidae) (McMullin et al. 2003). Vestimentiferans are important members of seep communities because they cluster in aggregations that provide living space for other fauna (Bergquist et al. 2003). Three described species of the genus Escarpia are distributed at seeps below 950 m along continental margins of the Atlantic and Pacific (Olu et al. 2010). Escarpia laminata occurs across the Gulf of
D. A. Cowart (&) C. Huang S. W. Schaeffer Department of Biology, The Pennsylvania State University, 208 Erwin W. Mueller Laboratory, University Park, PA 16801, USA e-mail: [email protected]
Mexico; Escarpia spicata is present in the Gulf of California, and Escarpia southwardae occurs along the equatorial margin of West Africa (Jones 1985; Andersen et al. 2004). Seep communities are under increased environmental threat from dumping, oil and gas drilling, so we employed 454-pyrosequencing to develop microsatellites for Escarpia to understand how these communities are established and maintained. Microsatellites are often used due to high rates of polymorphism, which can confirm population differentiation undetected by less sensitive analyses. Additionally, the use of next-generation sequencing reduces time and costs associated with traditional marker development methods (Allentoft et al. 2009). Furthermore, microsatellites can be applied across species to clarify phylogenetic relationships (Primmer et al. 2005; Hausdorf and Hennig 2010), and variation is expected to yield high returns for taxa within genera, with similar genome sizes and outcrossed mating systems (Barbara´ et al. 2007), all of which are characteristics of Escarpia. Here we report several novel microsatellites that are species s
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