EST-based microsatellites for northern pike ( Esox lucius ) and cross-amplification across all Esox species
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EST-based microsatellites for northern pike (Esox lucius) and cross-amplification across all Esox species G. Ouellet-Cauchon • E. Normandeau M. Mingelbier • L. Bernatchez
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Received: 1 December 2013 / Accepted: 25 December 2013 Ó Springer Science+Business Media Dordrecht 2014
Abstract Northern pike experience a global decline of populations and a better understanding of the species’ population genetic diversity and structure is needed for proper conservation and management. We developed 17 novel microsatellite markers in North American northern pike, Esox lucius, and tested cross-amplification on European populations as well as all five other Esox species. One marker deviated significantly from Hardy–Weinberg equilibrium and no linkage disequilibrium among all loci pairs was observed. A mean of 6.88 alleles per locus was found (between 2 and 23 alleles) and mean expected heterozygozity was 0.49 (range 0.033–0.950). All loci were successfully amplified on E. lucius from North America and Europe and E. reichertii. Between eight and 11 loci were successfully amplified in other Esox species and four out of 17 loci were successfully cross-amplified on all species. Keywords Esox lucius Northern pike EST-linked microsatellite Primers PCR
G. Ouellet-Cauchon (&) E. Normandeau L. Bernatchez Institut de Biologie Inte´grative et des Syste`mes (IBIS), Universite´ Laval, 1030 Avenue de la Me´decine, Quebec, QC G1V 0A6, Canada e-mail: [email protected] M. Mingelbier Service de la Faune Aquatique, Ministe`re du De´veloppement durable, de l’Environnement, de la Faune et des Parcs du Que´bec (MDDEFP), 880 chemin Sainte-Foy, Que´bec, QC G1S 4X4, Canada
Primer note Northern pike (Esox lucius) is a ubiquitous fish in the northern hemisphere’s freshwater bodies and is heavily exploited by recreational and commercial fisheries. Many stocks are also impacted by habitat alterations (Casselman and Lewis 1996). Northern pike populations have been shown to decline globally (e.g. Casselman and Lewis 1996; Smith et al. 2007). A better understanding of the species’ population genetic diversity and structure is needed if we are to achieve proper conservation and management. Towards this end we developed novel polymorphic microsatellite markers for this species. We developed 17 pairs of primers from EST sequences available from the cGRASP EST database (Table 1). Based on blast searches (Blast2Go software; Conesa et al. 2005), only EL21 was annotated and corresponded to the Danio rerio solute carrier organic anion transporter family member 2b1 gene. Microsatellite markers were amplified on 30 E. lucius individuals captured from three locations in St. Lawrence River in North America (46°240 200 °N, -72°220 800 °W; 45°220 1100 °N, -73°460 1900 °W; 44°250 1600 °N, -75°510 5500 °W). Cross-amplification was tested on all five Esox species [E. lucius from Europe (n = 20), E. reichertii (n = 17) from Eurasia, E. americanus americanus (n = 10), E. americanus vermiculatus (n = 10), E. masquinongy
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