Characterization and cross-amplification of microsatellite markers in four species of anemonefish (Pomacentridae, Amphiprion spp.)
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Anemonefish are iconic symbols of coral reefs and have become model systems for research on larval dispersal and population connectivity in coral reef fishes. Here we present 24 novel microsatellite markers across four species of anemonefish and also test 35 previously published markers for cross-amplification on two anemonefish species in order to facilitate further research on their population genetics and phylogenetics. Novel loci were isolated from sequences derived from microsatellite-enriched or 454 GS-FLX shotgun sequence libraries developed using congeneric DNA. Primer testing successfully identified 15 new microsatellite loci for A. percula, 4 for A. melanopus, 3 for A. akindynos, and 2 for A. omanensis. These novel microsatellite loci were polymorphic with a mean of 10 ± 1.6 SE (standard error) alleles per locus and an average observed heterozygosity of 0.647 ± 0.032 SE. Reliable cross-amplification of 12 and 26 of the 35 previously published Amphiprion markers was achieved for A. melanopus and A. akindynos, respectively, suggesting that the use of markers developed from the DNA of congeners can provide a quick and cost-effective alternative to the isolation of new loci. Together, the markers presented here provide an important resource for ecological, evolutionary, and conservation genetic research on anemonefishes that will inform broader conservation and management actions for coral reef fishes.
KeywordsAmphiprion Clownfish Coral reefs Microsatellite Population genetics
We are grateful to the many volunteers who assisted with sample collections, including Thomas Mannering, Rohan Brooker, Lisa Boström-Einarsson, Scott Hansen, Dylan Simonson, and Tessa Hempson. This project was supported by the King Abdullah University of Science and Technology (KAUST) (Awards CRG-1-2012-BER-002, OCRF-SPCF-2011-BER-001, and baseline research funds to MLB) and the Australian Government’s National Environmental Research Program (NERP) Tropical Ecosystems Hub (Project 8.3). Additional support was provided by the Australian Research Council (ARC) Centre of Excellence for Coral Reef Studies, the KAUST Red Sea Research Center, and the KAUST Biosciences Core Laboratory.
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