Isolation and characterization of polymorphic microsatellite markers from a globally distributed marine apex predator, t
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Isolation and characterization of polymorphic microsatellite markers from a globally distributed marine apex predator, the tiger shark (Galeocerdo cuvier) A. M. Bernard • K. A. Feldheim • M. S. Shivji
Received: 9 December 2014 / Accepted: 16 December 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract High levels of fishing have resulted in declines among many of the oceans top predators, including the globally distributed tiger shark (Galeocerdo cuvier). Overexploitation of this species has led to declines in parts of its distribution, which may have important ecological consequences given the tiger shark’s trophic position as a large bodied, generalist predator. To assess the population genetics of this species, an enrichment protocol was used to isolate a suite of nine di- and tetra-nucleotide microsatellite loci within the tiger shark, which were subsequently characterized using genomic DNA derived from 40 individuals sampled from Hawaiian waters. The number of alleles per locus ranged from 3 to 23, and the average expected heterozygosities across loci ranged from 0.30 to 0.93. Analyses suggested a low frequency of null alleles across markers, and all loci conformed to both Hardy– Weinberg and linkage equilibrium at P [ 0.01. Keywords Tiger shark Galeocerdo cuvier Microsatellite Elasmobranch genetics
Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0408-0) contains supplementary material, which is available to authorized users. A. M. Bernard M. S. Shivji (&) Save Our Seas Shark Research Center and Guy Harvey Research Institute, Nova Southeastern University, Oceanographic Center, 8000 North Ocean Drive, Dania Beach, FL 33004, USA e-mail: [email protected] K. A. Feldheim Pritzker Laboratory for Molecular Systematics and Evolution, The Field Museum of Natural History, 1400 South Lake Shore Drive, Chicago, IL 60605, USA
The tiger shark (Galeocerdo cuvier) is a globally distributed species that inhabits warm temperate and tropical coastal pelagic waters. As a large, generalist predator, the tiger shark has been identified as a keystone species within some marine ecosystems. The tiger shark is categorized as globally ‘Near Threatened’ on the IUCN Red List of Threatened Species (Simpfendorfer 2009), rendering it an important conservation priority. To date, little is known about the genetic connectivity, and demographic and evolutionary histories of this species, underscoring the need for useful genetic markers to study the population genetics of this important predator. Microsatellite markers were developed from genomic DNA (gDNA), extracted using the DNeasy Kit (QIAGEN, Valencia, CA), from a single tiger shark following the enrichment protocol of Glenn and Schable (2005) with some alterations: (1) restriction digest of gDNA was performed using the restriction enzymes RsaI and XmnI; and (2) two separate enrichments were performed: one using tetranucleotide probes [(AAAG)8, (ACAT)8, (AGAT)8, (AACT)8, (AAAT) 8, (AAGT)8] and one usi
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