Abstract
As a first step to establish the genetic structure of the sea urchin Echinometra lucunter lucunter throughout the Caribbean Sea, 26 microsatellite loci were isolated using Illumina paired-end sequencing, Next Generation Sequencing (NGS). We successfully optimized 17 loci for genotyping and the variation tested for 23 individuals from the Caribbean Sea and Tropical Eastern Atlantic Ocean. The allele number per locus (Na) ranged from four to 24, the observed heterozygosity (Ho) from 0.682 to 1, and the expected heterozygosity (He) from 0.609 to 0.9304. We detected no linkage disequilibrium between pairs of loci. These microsatellites will be used for the first time to detect the influence of marine barriers to genetic flow in the sea urchin E. lucunter lucunter throughout the Caribbean Sea. These new validated markers will be essential for current conservation and connectivity studies across the Caribbean Sea and the Atlantic Ocean.
References
Avise, J. C. (1992). Molecular population structure and biogeographic history of a regional fauna a case history with lessons for conservative biology. Oikos. 63: 62-76. Doi: 10.2307/3545516
Baums, I. B., Miller, M. W., Hellberg, M. E. (2005). Regionally isolated populations of an imperiled Caribbean coral, Acropora palmata. Mol Ecol. 14: 1377-1390. Doi: 10.1111/j.1365-294X.2005.02489.x
Blacket, M. J., Robin. C., Good, R. T, Lee, S. F, Miller, A. D. (2012). Universal primers for fluorescent labelling of PCR fragments – an effective approach to genotyping by fluorescence. Mol Ecol Resour. 1: 456-463. Doi: 10.1111/j.1755-0998.2011.03104.x
Carlin, J. L., Robertson, D. R., Bowen, B. W. (2003). Ancient divergences and recent connections in two tropical Atlantic reef fishes Epinephelus adscensionis and Rypticus saponaceous (Percoidei: Serranidae). Mar Biol. 143: 1057-1069. Doi: 10.1007/s00227-003-1151-3
Cowen, R. K., Paris, C. B., Srinivasan, A. (2006). Scaling of population connectivity in marine populations. Sci. 311: 522-527. Doi: 10.1126/science.1122039
Culley, T. M., Stamper, T. I., Stokes, R. L., Brzski, J. R., Hardman, N. A., Klooster, M. R, Merritt, B. J. (2013). An efficient technique for primer development and application that integrates fluorescent labelling and multiplex PCR. Appl Plant Sci. 1 (10): 1-10. Doi: 10.3732/apps.1300027
Fox, G., Preziosi, R. F., Antwis, R. E., Benavides‐Serrato M., Combe, F. J., Harris, W. E., Hartley, I. R., Kitchener, A. C., De Kort, S. R., Nekaris, A. I., Rowntree, J. K. (2019). Multi‐individual microsatellite identification: A multiple genome approach to microsatellite design (MiMi). Mol Ecol Resour. 19 (6): 1-9. Doi: 10.1111/1755-0998.13065
Griffiths, S. M., Fox, G., Briggs, P. J., Donaldson, I. J., Hood, S., Richardson, P., Leaver, G. W., N. K. Truelove, Preziosi, R. F. (2016). A Galaxy-based bioinformatics pipeline for optimised, streamlined microsatellite development from Illumina next-generation sequencing data. Conserv Genet Resour. 8: 481-486. Doi: 10.1007/s12686-016-0570-7
Kalinowski, S. T., Taper, M. L., Marshall, T.C. (2007). Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Mol Ecol. 16: 1099-1106. Doi: 10. 1111/j.1365-294X.2007.03089.x
McCartney, M. A., Keller, G., Lessios, H. A. (2000). Dispersal barriers in tropical oceans and speciation in Atlantic and eastern Pacific sea urchins of the genus Echinometra. Mol. Ecol. 9: 1391-1400. Doi: 10.1046/j.1365-294x.2000.01022.x
Paris, C. B., Cowen, R. K. (2004). Direct evidence of a biophysical retention mechanism for coral reef fish larvae. Limnol. 49 (6): 1964-1979. Doi: 10.4319/lo.2004.49.6.1964
Pawson, D. L. (1978). The echinoderm fauna of Ascension Island, South Atlantic Ocean. Smithsonian Contrib. Mar. Sci, 2: 1-29.
Qiagen. DNeasy® Blood & Tissue Handbook [pdf] (2006). Date of entry: January 15, 2015. Available in: http://mvz.berkeley.edu/egl/inserts/DNeasy_Blood_&_Tissue_Handbook.pdf
Qiagen. Type-it Microsatellite PCR Handbook [pdf] (2009). Date of entry: March 14, 2015. Available in: http://www.qiagen.com/Products/Catalog/Assay-Technologies/End-Point-PCR-and-RTPCR-Reagents/Type-it-Microsatellite-PCR-Kit#resources
Schultz, E. T. & Cowen, R. K. (1994). Recruitment of coral-reef fishes to Bermuda – local retention or long-distance transport. Mar Ecol Prog Ser. 109: 15-28. Doi: 10.3354/meps111015
Selkoe, K. A. & Toonen, R. J. (2006). Microsatellites for ecologists: A practical guide to using and evaluating microsatellite markers. Ecol Lett. 9: 615-29. Doi: 10.1111/j.1461-0248.2006.00889.x
Sunnucks, P. (2000). Efficient genetic markers for population biology. Trends Ecol Evol. 15 (5): 199-203. Doi: 10.1016/S0169-5347(00)01825-5
Taylor, M. S. & Hellberg, M. E. (2003). Genetic evidence for local retention of pelagic larvae in a Caribbean reef fish. Sci. 299: 107-109. Doi: 10.1126/science.1079365
Taylor, M. S. & Hellberg, M. E. (2006). Comparative phylogeography in a genus of coral reef fishes: biogeographic and genetic concordance in the Caribbean. Mol Ecol. 15: 695-707. Doi: 10.1111/j.1365-294X.2006.02820.x
Van Oosterhout, C., Hutchison, W. F., Shipley, P., Wills, D. P. M. (2004). Micro-Checker: Software for identifying and correcting genotyping errors in microsatellite data. Mol. Ecol. Notes. 4: 535-538. Doi: 10.1111/j.1471-8286.2004.00684.x
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Copyright (c) 2020 Revista de la Academia Colombiana de Ciencias Exactas, Físicas y Naturales