Novel polymorphic microsatellite loci for the eastern king prawn, Penaeus (Melicertus) plebejus
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MICROSATELLITE LETTERS
Novel polymorphic microsatellite loci for the eastern king prawn, Penaeus (Melicertus) plebejus Jackie T. Chan • Sharon A. Appleyard • William B. Sherwin • Matthew D. Taylor
Received: 20 May 2013 / Accepted: 30 May 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract Eastern king prawn, Penaeus (Melicertus) plebejus, is an endemic Australian species which is heavily exploited both commercially and recreationally in estuaries and in offshore trawl fisheries across several jurisdictions. P. plebejus shows extensive movement throughout the known distribution range during various life history stages (Montgomery et al. in Fish Res 80:80–87, 2007). It is currently unknown whether there is a full migration throughout the species range, or step-wise migration, and identifying the origins of important spawning populations remains as a challenge. We report 20 high resolution
Electronic supplementary material The online version of this article (doi:10.1007/s12686-013-9972-y) contains supplementary material, which is available to authorized users. J. T. Chan S. A. Appleyard CSIRO Food Futures National Research Flagship, 5 Julius Avenue, North Ryde, NSW 2113, Australia J. T. Chan CSIRO Livestock Industries, Queensland Biosciences Precinct, 306 Carmody Road, St. Lucia, QLD 4067, Australia J. T. Chan S. A. Appleyard CSIRO Marine and Atmospheric Research, 233 Middle Street, Cleveland, QLD 4163, Australia J. T. Chan W. B. Sherwin M. D. Taylor Evolution and Ecology Research Centre, School of Biological Earth and Environmental Science, University of New South Wales, Sydney, NSW 2052, Australia M. D. Taylor (&) Wild Fisheries Research, Port Stephens Fisheries Institute, NSW Department of Primary Industries, Locked Bag 1, Nelson Bay, NSW 2315, Australia e-mail: [email protected]
microsatellite markers, to address such questions and assist conservation planning for long-term sustainability. Keywords Penaeidae Stock enhancement Population structure
Microsatellite note Microsatellites were isolated using enrichment library and Roche 454 pyrosequencing approaches. Genomic DNA of five individuals were used for constructing the enriched libraries targeting four motifs (ACAT8, GGA8, AGAT8 and AAAC8) using biotin-labelled micro-beads. Cloning was done using pGEM-T vectors and One Shot TOP10 competent cells. Recombinant clones were selected by standard bluewhite-screening and subsequently sequenced. Approximately 75 % of the screened clones contained detectable repeat structure (Table 1). Only the GGA8 motif showed a high level of redundancy. Forty-two candidate loci were screened by PCR and high-resolution 3 % agarose gel using a test panel (seven individuals), polymorphism was determined by the number of bands. Fifteen loci were selected for fluorescence labelling based on: (1) success and specificity of amplification; and (2) level of polymorphism. A final eight were subsequently put into two multiplexes and characterised using 27 individuals. Locus L4E07 was characterised as single
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