Abstract
Extensive studies of genetic variation of Atlantic horseshoe crab Limulus polyphemus populations have revealed the presence of considerable allelic diversity and population structuring that appear to reflect the actions of various evolutionary processes. We have expanded on our previous efforts to gain a more refined understanding of L. polyphemus population structure by surveying 792 additional animals distributed among 12 additional spawning aggregations. Here we report on variation at 13 microsatellite DNA markers for 1,684 horseshoe crabs sampled from 33 spawning assemblages from northern Maine to the Yucatan Peninsula, Mexico. Average unbiased heterozygosity (uH E ) was high (0.74 ± 0.01), the number of private alleles was low (0.06 ± 0.04), effective population size (N e) ranged from 22 to 187, inbreeding (F) ranged from −0.07 to 0.07, and tests for genic differentiation among populations indicated shallow but statistically significant differentiation within regions and highly significant differences among regions (P < 0.005). Current findings are consistent with previous research by this group in suggesting a series of genetic discontinuities across the species’ range that could indicate regional adaptive significance or reflect vicariant geographic events. Additional collections allowed improved delineation of structuring (as reflected by two new zones of genetic discontinuity) along the southeast Atlantic coast as well as identification of previously undetected shallow but significant structuring along the Florida Gulf coast. Regional groupings may warrant management unit recognition based on the patterns observed among multiple genetic metrics. The integration of this information with previously identified genetic variation and ecological data is essential to developing an ecologically and evolutionarily sound conservation management strategy.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Allendorf FW, Luikart G (2007) Conservation and the genetics of populations. Blackwell Publishing, Malden
ASMFC (Atlantic States Marine Fisheries Commission) (1998) Interstate fishery management plan for horseshoe crab. ASMFC, Fishery Management report no. 32, Washington, DC
Avise JC (1992) Molecular population structure and the biogeographic history of a regional fauna: a case history with lessons for conservation biology. Oikos 63:62–76
Baker AJ, Gonzalez PM, Piersma T et al (2004) Rapid population decline in red knots: fitness consequences of decreased refueling rates and late arrival in Delaware Bay. Proc R Soc Lond B Biol Sci 271:875–882
Barton NH, Slatkin M (1986) A quasi-equilibrium theory of the distribution of rare alleles in a subdivided population. Heredity 56:409–415
Bernatchez L, Wilson CC (1998) Comparative phylogeography of Nearctic and Palearctic fishes. Mol Ecol 7:431–452
Botton ML, Loveland RE (2003) Abundance and dispersal potential of horseshoe crab (Limulus polyphemus) larvae in the Delaware estuary. Estuaries 26:1472–1479
Brockmann HJ, Colson T, Potts W (1994) Sperm competition in horseshoe crabs (Limulus polyphemus). Behav Ecol Sociobiol 35:153–160
Brockmann HJ, Nguyen C, Potts W (2000) Paternity in horseshoe crabs when spawning in multiple male groups. Anim Behav 60:837–849
Cavalli-Sforza LL, Edwards AWF (1967) Phylogenetic analysis: models and estimation procedures. Evolution 21:550–570
Cornuet J-M, Piry S, Luikart G et al (1999) New methods employing multilocus genotypes to select or exclude populations as origins of individuals. Genetics 153:1989–2000
Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet Resour 4(2):359–361
Ehlinger GS, Tankersley RA (2004) Survival and development of horseshoe crab (Limulus polyphemus) embryos and larvae in hypersaline conditions. Biol Bull 206:87–94
Ehlinger GS, Tankersley RA (2007) Reproductive ecology of the American horseshoe crab Limulus polyphemus in the Indian River Lagoon: an overview. Fla Sci 70:449–463
Ehlinger GS, Tankersley RA (2009) Ecology of horseshoe crabs in microtidal lagoons. In: Tanacredi JT, Botton M, Smith DR (eds) Biology and conservation of horseshoe crabs. Springer, New York, pp 149–162
Ellegren H (2004) Microsatellites: simple sequences with complex evolution. Nat Rev Genet 5:435–455
Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620
Excoffier L, Smouse PE, Quattro JM (1992) Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics 131:479–491
Faurby S, King TL, Obst M et al (2010) Population dynamics of American horseshoe crabs–historic climatic events and recent demographic pressures. Mol Ecol 19:3088–3100
Fulford RS, Haehn RA (2012) An evaluation of Mississippi barrier islands as spawning and nesting habitat for the American horseshoe crab, Limulus polyphemus, with implications for island restoration. Gulf Caribb Res 24:51–62
Gerhart SD (2007) A review of the biology and management of horseshoe crabs with emphasis on Florida populations. Fish and Wildlife Research Institute, St. Petersburg, http://research.myfwc.com. Accessed Jan 2015
Gomez-Aguirre S (1993) Cacerolita de Mar (Limulus polyphemus L.) en la Peninsula de Yucatan. In: Salazar-Vallejo SI, Gonzalez NE (eds) Biodiversidad Marina ye Costera de Mexico. Com. Nal. Biodiversidad y CIQRO, Mexico, D.F., pp 650–659
Goudet J, Perrin N, Waser P (2002) Tests for sex-biased dispersal using bi-parentally inherited genetic markers. Mol Ecol 11:1103–1114
Graham LJ, Botton ML, Hata D et al (2009) Prosomal-width-to-weight relationships in American horseshoe crabs (Limulus polyphemus): examining conversion factors used to estimate landings. Fish Bull 107:235–243
Guo SW, Thompson EA (1992) Performing the exact test of Hardy-Weinberg proportions for multiple alleles. Biometrics 48:361–372
Hallerman EM (2003) Population genetics: principles and applications for fisheries scientists. American Fisheries Society, Bethesda
Hammer Ø, Harper DAT, Ryan PD (2001) Past: paleontological statistics software package for education and data analysis. Palaeontol Electron 4(1):9pp, art. 4, 178kb. http://palaeoelectronica.org/2001_1/past/issue1_01.htm
Hastings A (1993) Complex interactions between dispersal and dynamics–lessons from coupled logistic equations. Ecology 74:1362–1372
Höss M, Pääbo S (1993) DNA extraction from Pleistocene bones by a silica-based purification method. Nucleic Acids Res 21:3913–3914
King TL, Eackles MS, Spidle AP et al (2005) Regional differentiation and sex-biased dispersal among populations of the horseshoe crab Limulus polyphemus. Trans Am Fish Soc 134:441–465
King TL, Switzer JF, Morrison CL et al (2006) Comprehensive genetic analyses reveal evolutionary distinction of a mouse (Zapus hudsonius preblei) proposed for delisting from the US Endangered Species Act. Mol Ecol 15:4331–4359
Levin J, Bang FB (1968) Clottable protein in Limulus: its localization and kinetics of its coagulation by endotoxin. Thromb Diath Haemorrh 19(1):186–197
Meirmans PG (2006) Using the AMOVA framework to estimate a standardized genetic differentiation measure. Evolution 60:2399–2402
Moritz C (1994) Defining ‘evolutionary significant units’ for conservation. Trends Ecol Evol 9:373–375
Paetkau D, Calvert W, Stirling I et al (1995) Microsatellite analysis of population structure in Canadian polar bears. Mol Ecol 4:347–354
Palsbøll PJ, Berube M, Allendorf FW (2007) Identification of management units using population genetic data. Trends Ecol Evol 22:11–16
Peakall ROD, Smouse PE (2006) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol Ecol Notes 6:288–295
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update. Bioinformatics 28:2537–2539
Pierce JP, Tan G, Gaffney PM (2000) Delaware Bay and Chesapeake Bay populations of the horseshoe crab (Limulus polyphemus) are genetically distinct. Estuaries 23:690–698
Piry S, Alapetite A, Cornuet J-M et al (2004) GeneClass2: a software for genetic assignment and first-generation migrant detection. J Hered 95:536–539
Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959
Provancha J (1997) Annual report for sea turtle netting in Mosquito Lagoon. NMFS Permit #942, FL Permit #114
Ramilo ST, Wang J (2012) The effect of close relatives on unsupervised Bayesian clustering algorithms in population genetic structure analysis. Mol Ecol Resour 12:873–884
Ramstad KM, Woody CA, Sage GK et al (2004) Founding events influence genetic population structure of sockeye salmon (Oncorhynchus nerka) in Lake Clark, Alaska. Mol Ecol 13:277–290
Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249
Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225
Riska B (1981) Morphological variation in the horseshoe crab Limulus polyphemus. Evolution 35:647–658
Rohlf FJ (2000) NTSYS-PC: numerical taxonomy and multivariate analysis systems, Version 2.10. Exeter Software, Setauket
Ryder O (1986) Species conservation and systematics: the dilemma of subspecies. Trends Ecol Evol 1:9–10
Saunders NC, Kessler LG, Avise JC (1986) Genetic variation and geographic differentiation in mitochondrial DNA of the horseshoe crab, Limulus polyphemus. Genetics 112:613–627
Scheidt D, Lowers R (2001) Using an aerial survey to document the extent of a horseshoe crab die-off in Florida. In: American Fisheries Society annual meeting, Baltimore, MD
Selander RK, Yang SY, Lewontin RC et al (1970) Genetic variation in the horseshoe crab (Limulus polyphemus), a phylogenetic “relic”. Evolution 24:402–414
Shuster CN Jr (1979) Distribution of the American horseshoe “crab”, Limulus polyphemus (L.). In: Cohen E (ed) Biomedical applications of the horseshoe crab (Limulidae). Alan R. Liss, Inc., New York, pp 3–26
Shuster CS, Barlow RB, Brockmann HJ (eds) (2003) The American horseshoe crab. Harvard University Press, Cambridge, MA
Sokal RR, Rohlf FJ (1994) Biometry: the principles and practice of statistics in biological research, 3rd edn. W.H. Freeman & Co., New York
Swofford DL, Selander RB (1981) BIOSYS-1: A FORTRAN program for the comprehensive analysis of electrophoretic data in population genetics and systematics. J Hered 72:281–283
Walls EA, Berkson J, Smith SA (2002) The horseshoe crab, Limulus Polyphemus: 200 million years of existence, 100 years of study. Rev Fish Sci 10:39–73
Wang J, Santure AW (2009) Parentage and sibship inference from multilocus genotype data under polygamy. Genetics 181:1579–1594
Wright S (1931) Evolution in Mendelian populations. Genetics 16:97–159
Zaldivar-Rae J, Sapien-Silva RE, Rosales-Raya M et al (2009) American horseshoe crabs, Limulus polyphemus, in Mexico: open possibilities. In: Tanacredi JT, Botton M, Smith DR (eds) Biology and conservation of horseshoe crabs. Springer, New York, pp 97–113
Acknowledgements
The U.S. Geological Survey (State Partnership and Quick Response programs), the Atlantic States Marine Fisheries Commission, and the AAAS Women’s International Science Collaboration Program funded this research. The following individuals graciously provided L. polyphemus tissue samples: S. Schaller, C. McBane, K. Tuxbury, C. Grahn, B. Battelle, M-J. James-Pirri, T. O’Connell, J. Mattei, R. Chapman, G. Ehlinger, L. Barton, T. Summers, C. Callahan, C. Morrison, D.R. Smith, K. Voges, A. Garcia, Z. Johnson, R. Johnson, K. Hovatter, V. Robbins, and P. Pooler. Assistance with the Yucatan field collection was provided by G. V. Rios, Centro Regional de Investigación Pesquera, Yucalpetén, Mexico. We thank the State of Florida Fish and Wildlife Conservation Commission and Ryan Gandy and Michael Tringali (St. Petersburg Office) for their kind help in providing genotype data on the newest Florida collections. Laboratory assistance was provided by B. Eleby, R. Long, R. Johnson, and M. Meadows. We thank J. Young for assistance in generating the collection site map. Funding for EH’s participation in this work was provided in part by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture. Use of trade, product, or firm names does not imply endorsement by the U.S. Government.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer International Publishing Switzerland
About this chapter
Cite this chapter
King, T.L., Eackles, M.S., Aunins, A.W., Brockmann, H.J., Hallerman, E., Brown, B.L. (2015). Conservation Genetics of the American Horseshoe Crab (Limulus polyphemus): Allelic Diversity, Zones of Genetic Discontinuity, and Regional Differentiation. In: Carmichael, R., Botton, M., Shin, P., Cheung, S. (eds) Changing Global Perspectives on Horseshoe Crab Biology, Conservation and Management. Springer, Cham. https://doi.org/10.1007/978-3-319-19542-1_4
Download citation
DOI: https://doi.org/10.1007/978-3-319-19542-1_4
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-19541-4
Online ISBN: 978-3-319-19542-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)