Advertisement

Conservation Genetics

, 9:1263 | Cite as

Patterns of nuclear and mitochondrial DNA variation in Iberian populations of Emys orbicularis (Emydidae): conservation implications

  • Guillermo Velo-Antón
  • Mario García-París
  • Adolfo Cordero Rivera
Research Article

Abstract

The European pond turtle (Emys orbicularis) is threatened and in decline in several regions of its natural range, due to habitat loss combined with population fragmentation. In this work, we have focused our efforts on studying the genetic diversity and structure of Iberian populations with a fine-scale sampling (254 turtles in 10 populations) and a representation from North Africa and Balearic island populations. Using both nuclear and mitochondrial markers (seven microsatellites, ∼1048 bp nDNA and ∼1500 bp mtDNA) we have carried out phylogenetic and demographic analyses. Our results show low values of genetic diversity at the mitochondrial level although our microsatellite dataset revealed relatively high levels of genetic variability with a latitudinal genetic trend decreasing from southern to northern populations. A moderate degree of genetic differentiation was estimated for Iberian populations (genetic distances, F ST values and clusters in the Bayesian analysis). The results in this study combining mtDNA and nDNA, provide the most comprehensive population genetic data for E. orbicularis in the Iberian Peninsula. Our results suggest that Iberian populations within the Iberian–Moroccan lineage should be considered as a single subspecies with five management units, and emphasize the importance of habitat management rather than population reinforcement (i.e. captive breeding and reintroduction) in this long-lived species.

Keywords

Emys orbicularis Genetic diversity mtDNA nDNA Population structure 

Notes

Acknowledgements

We thank all the people who helped us to collect samples: César Ayres from most populations; Pedro Segurado from Portugal; Gonzalo Alarcos from Salamanca and Zamora; Judith Hidalgo, Matati, Alex Portheault and Ana Andreu from the Doñana National Park; Vicente Sancho and Ignacio Lacomba from Valencia; Alberto Álvarez from Madrid, Carlos Gómez Lorenzo from Ourense. Alain Veysset provided samples from Menorca; David Donaire and Salvador Carranza supplied samples from Morocco. David Buckley, Iñigo Martínez-Solano and three anonymous reviewers made helpful comments and suggestions on the manuscript. Christopher Beatty improved English on the manuscript. GV-A is supported by a grant from Galician Government (Conselleria de Innovación, Industria e Comercio, Xunta de Galicia). Funding was provided by Research grants from Xunta de Galicia (PGIDT01MAM37101PR, PGIDIT05RFO37101PR) and Cátedra Filgueira Valverde-Universidade de Vigo to ACR.

References

  1. Alexandrino J, Froufe E, Arntzen JW, Ferrand N (2001) Genetic subdivision, glacial refugia and postglacial recolonization in the golden-striped salamander, Chioglossa lusitanica (Amphibia:urodela). Mol Ecol 9:771–781CrossRefGoogle Scholar
  2. Altherr S, Freyer D (2000) Asian turtles are threatened by extinction. Turtle Tortoise Newsl 1:7–11Google Scholar
  3. Avise JC, Bowen BW, Lamb T, Meylan AB, Bermingham E (1992) Mitochondrial-DNA a evolution at a turtles pace – evidence for low genetic variability and reduced microevolutionary rate in the testudines. Mol Biol Evol 9:457–473PubMedGoogle Scholar
  4. Ayres Fernández C, Cordero Rivera A (2004) The incidence of asymmetries and accessory plates in Emys orbicularis from NW Spain. Biologia 14(59/Suppl):85–88Google Scholar
  5. Beheregaray LB, Ciofi C, Caccone A, Gibbs JP, Powell JR (2003) Genetic divergence, phylogeography and conservation units of giant tortoises from Santa Cruz and Pinzon, Galapagos Islands. Conserv Genet 4:31–46CrossRefGoogle Scholar
  6. Belkhir K, Borsa P, Chikhi L, Raufaste N, Bonhomme F (2004) GENETIX 4.05, logiciel sous Windows TM pour la génétique des populations. Laboratoire Génome, Populations, Interactions, MontpellierGoogle Scholar
  7. Bertolero A (2000) Suivi de la population de Cistude Emys orbicularis dans le delta de l’Ebre (NE Espagne). Chelonii 2:63–68Google Scholar
  8. Bishop CA, Ng P, Pettit KE, Kennedy SW, Stegeman JJ, Norstrom RJ, Brooks RJ (1998) Environmental contamination and developmental abnormalities in eggs and hatchlings of the common snapping turtle (Chelydra serpentina serpentina) from the Great Lakes-St. Lawrence River basin (1989–91). Environ Pollut 101:143–156PubMedCrossRefGoogle Scholar
  9. Cabral JM, Magalhães P, Oliveira E, Romão C (1990) Livro vermelho dos vertebrados de Portugal. Vol. I – Mamíferos, aves, répteis e anfibios. Serviço Nacional de Parques, Reservas e Conservaçao da Natureza, LisboaGoogle Scholar
  10. Caccone A, Gentile G, Burns CE, Sezzi E, Bergman W, Ruelle M, Saltonstall K, Powell JR (2004) Extreme difference in rate of mitochondrial and nuclear DNA evolution in a large ectotherm, Galapagos tortoises. Mol Phylogenet Evol 31:794–798PubMedCrossRefGoogle Scholar
  11. Carranza S, Arnold EN (2004) History of West Mediterranean newts, Pleurodeles (Amphibia: Salamandridae), inferred from old and recent DNA sequences. Syst Biodivers 1(3):327–337CrossRefGoogle Scholar
  12. Carranza S, Arnold EN, Wade E, Fahd S (2004) Phylogeography of the false smooth snakes, Macroprotodon (Serpentes, Colubridae): mitochondrial DNA sequences show European populations arrived recently from Northwest Africa. Mol Phylogenet Evol 33:523–532PubMedCrossRefGoogle Scholar
  13. Carranza S, Arnold EN, Pleguezuelos JM (2006) Phylogeny, biogeography, and evolution of two Mediterranean snakes, Malpolon monspessulanus and Hemorrhois hippocrepis (Squamata, Colubridae), using mtDNA sequences. Mol Phylogenet Evol 40:532–546PubMedCrossRefGoogle Scholar
  14. Cavallis LL, Edwards AWF (1967) Phylogenetic analysis models and estimation procedures. American Journal of Human Genetics 19:233Google Scholar
  15. Clement M, Posada D, Crandall KA (2000) TCS: a computer program to estimate gene genealogies. Mol Ecol 9:1657–1659PubMedCrossRefGoogle Scholar
  16. Congdon JD, Dunham AE, Sels RCV (1993) Delayed sexual maturity and demographics of Blanding´s turtles (Emydoidea blandingii) – implications for conservation and management of long-lived organisms. Conserv Biol 7:826–833CrossRefGoogle Scholar
  17. Cordero Rivera A, Ayres Fernández C (2004) A management plan for the European pond turtle (Emys orbicularis) populations of the river Louro basin (NW Spain). Biologia 14(59/Suppl):161–171Google Scholar
  18. Cournet JM, Luikart G (1997) Description and power analysis of two tests for detecting recent population bottlenecks from allele frecuency data. Genetics 144:2001–2014Google Scholar
  19. Cunningham J, Baard EHW, Harley EH, O’Ryan C (2002) Investigation of genetic diversity in fragmented geometric tortoise (Psammobates geometricus) populations. Conserv Genet 3:215–223CrossRefGoogle Scholar
  20. Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS (1991) Touchdown PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19:4008PubMedCrossRefGoogle Scholar
  21. Epperson DM, Heise CD (2003) Nesting and hatchling ecology of gopher tortoises (Gopherus polyphemus) in southern Mississippi. J Herpetol 37:315–324CrossRefGoogle Scholar
  22. Felsenstein J (1995) PHYLIP. [3.57c]. Department of Genetics, University of Washington, SeattleGoogle Scholar
  23. Filatov DA (2002) PROSEQ: a software for preparation and evolutionary analysis of DNA sequence data sets. Mol Ecol Notes 2:621–624CrossRefGoogle Scholar
  24. Forman RTT, Alexander LE (1998) Roads and their major ecological effects. Annu Rev Ecol Syst 29:207–231CrossRefGoogle Scholar
  25. Frankham R (1995) Conservation genetics. Annu Rev Genet 29:305–327PubMedCrossRefGoogle Scholar
  26. Frankham R, Ralls K (1998) Conservation biology: inbreeding leads to extinction. Nature 392:441–442CrossRefGoogle Scholar
  27. Frankham R, Ballou JD, Briscoe DA (2002) Introduction to conservation genetics. Cambridge University Press, CambridgeGoogle Scholar
  28. Friedel RH, Stubbusch J, Barde YA, Schnurch H (2001) A novel 7-transmembrane receptor expressed in nerve growth factor-dependent sensory neurons. Mol Cell Neurosci 17:31–40PubMedCrossRefGoogle Scholar
  29. Fritz U (1993) On the intraspecific variation of Emys orbicularis (Linnaeus, 1758). 3. Two new subspecies from the Iberian Peninsula and North Africa, Emys orbicularis fritzjuergenobsti.nov. and E.o.occidentalis subsp.nov. (Reptilia, Testudines: Emydidae). Zool Abh Mus Tierdk Dresden 47:131–155Google Scholar
  30. Fritz U (2001) Emys orbicularis (Linnaeus, 1758) – Europäische Sumpfschildkröte. In: Fritz U (ed) Handbuch der Reptilien und Amphibien Europas. Band 3/IIIA: Schildkröten I. Aula-Verlag, Wiebelsheim, pp 343–515Google Scholar
  31. Fritz U, Keller C, Budde M (1996) A new subspecies of the European pond turtle from southwestern Spain, Emys orbicularis hispanica subsp.nov. Salamandra 32:129–152Google Scholar
  32. Fritz U, Pieh A, Lenk P, Mayol J, Sättele B, Wink M (1998) Is Emys orbicularis introduced to Majorca? Mertensiella 10:122–133Google Scholar
  33. Fritz U, Barata M, Busack SD, Fritzsch G, Castilho R (2006) Impact of mountain chains, sea straits and peripheral populations on genetic and taxonomic structure of a freshwater turtle, Mauremys leprosa (Reptilia, Testudines, Geoemydidae). Zool Scr 35:97–108CrossRefGoogle Scholar
  34. Fritz U, Guicking D, Kami H et al (2007). Mitochondrial phylogeography of European pond turtle (Emys orbicularis, Emys trinacris) – an update. Amphibia-Reptilia 28:418–426CrossRefGoogle Scholar
  35. Fujita MK, Engstrom TN, Starkey DE, Shaffer HB (2004) Turtle phylogeny: insights from a novel nuclear intron. Mol Phylogenet Evol 31:1031–1040PubMedCrossRefGoogle Scholar
  36. Gaggiotti OE, Lange O, Rassmann K, Gliddon C (1999) A comparison of two indirect methods for estimating average levels of gene flow using microsatellite data. Mol Ecol 8:1513–1520PubMedCrossRefGoogle Scholar
  37. Galán P (1999) Conservación de la herpetofauna gallega. Universidade da Coruña, A CoruñaGoogle Scholar
  38. Gasith A, Sidis I (1984) Polluted water bodies, the main habitat of the Caspian terrapin (Mauremys caspica rivulata) in Israel. Copeia 1984:216–219CrossRefGoogle Scholar
  39. Gemmell NJ, Akiyama S (1996) An efficient method for the extraction of DNA from vertebrate tissues. Trends Genet 12:338–339PubMedCrossRefGoogle Scholar
  40. Gibbs JP, Shriver WG (2002) Estimating the effects of road mortality on turtle populations. Conserv Biol 16:1647–1652CrossRefGoogle Scholar
  41. Godinho R, Paulo OS, Ferrand N, Luis C, Rosa HD, Crespo EG (2003) Major patterns of population differentiation in the Iberian Schreiber’s green lizard (Lacerta schreiberi) inferred from protein polymorphism. Herpetol J 13:35–42Google Scholar
  42. Gómez A, Lunt DH (2007) Refugia within refugia: patterns of phylogeographic concordance in the Iberian Península. In: Weiss S, Ferrand N (eds) Phylogeography in southern European refugia: evolutionary perspectives on the origins and conservation of European biodiversity. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 155–188Google Scholar
  43. Gómez-Cantarino A, Lizana M (2000) Distribución y uso del hábitat de los galápagos (Mauremys leprosa y Emys orbicularis) en la provincia de Salamanca. Bol Asoc Herpetol Esp 11:4–8Google Scholar
  44. Goodman SJ (1997) RSTCALC: a collection of computer programs for calculating estimates of genetic differentiation from microsatellite data and determining their significance. Mol Ecol 6:881–885CrossRefGoogle Scholar
  45. Guo SW, Thomson EA (1992) Performing the exact test of Hardy-Weinberg proportion for multiple alleles. Biometrics 48:361–372PubMedCrossRefGoogle Scholar
  46. Hanski IA, Gilpin ME (1997) Metapopulation biology: ecology, genetics and evolution. Academic Press, San DiegoGoogle Scholar
  47. Harrison S, Hastings A (1996) Genetic and evolutionary consequences of metapopulation structure. Trends Ecol Evolut 11:180–183CrossRefGoogle Scholar
  48. Hart D, Clark A (1997) Principles of Population Genetics, 3rd edn. Sinauer Associates, SunderlandGoogle Scholar
  49. Hauswaldt JS, Glenn TC (2005) Population genetics of the diamondback terrapin (Malaclemys terrapin). Mol Ecol 14:723–732PubMedCrossRefGoogle Scholar
  50. Hedrick PW, Cockerham CC (1986) Partial inbreeding: equilibrium heterozygosity and the heterozygosity paradox. Evolution 40:856–861CrossRefGoogle Scholar
  51. Hewitt GM (1999) Post-glacial re-colonization of European biota. Biol J Linn Soc 68:87–112CrossRefGoogle Scholar
  52. Hewitt G (2000) The genetic legacy of the Quaternary ice ages. Nature 405:907–913PubMedCrossRefGoogle Scholar
  53. Janzen FJ, Hoover SL, Shaffer HB (1997) Molecular phylogeography of the western pond turtle (Clemmys marmorata): preliminary results. Chelelonian Conserv Biol 2:623–626Google Scholar
  54. Joyal LA, McCollough M, Hunter ML (2001) Landscape ecology approaches to wetland species conservation: a case study of two turtle species in southern Maine. Conserv Biol 15:1755–1762CrossRefGoogle Scholar
  55. Keller C, Andreu AC (2002) Emys orbicularis (Linnaeus, 1758). Galápago europeo. In: Pleguezuelos JM, Márquez R, Lizana M (eds) Atlas y Libro Rojo de los anfibios y reptiles de España. Ministerio de Medio Ambiente, Madrid, pp 181–186Google Scholar
  56. King TL, Julian SE (2004) Conservation of microsatellite DNA flanking sequence across 13 Emydid genera assayed with novel bog turtle (Glyptemys muhlenbergii) loci. Conserv Genet 5:719–725CrossRefGoogle Scholar
  57. Kuo C-H, Janzen FJ (2004) Genetic effects of a persistent bottleneck on a natural population of ornate box turtles (Terrapene ornata). Conserv Genet 5:425–437CrossRefGoogle Scholar
  58. Lenk P, Fritz U, Joger U, Winks M (1999) Mitochondrial phylogeography of the European pond turtle, Emys orbicularis (Linnaeus 1758). Mol Ecol 8:1911–1922PubMedCrossRefGoogle Scholar
  59. Machordom A, Doadrio I (2001) Evidence of a cenozoic Belt-Kabilian connection based on freshwater fish phylogeography (Luciobarbus, Cyprinidae). Mol Phylogent Evol 18:252–263CrossRefGoogle Scholar
  60. Martínez-Silvestre A, Perpinan D, Marco I, Lavin S (2002) Venipuncture technique of the occipital venous sinus in freshwater aquatic turtles. J Herpetol Med Surg 12(4):31–32Google Scholar
  61. Martinez-Solano I, Teixeira J, Buckley D, Garcia-Paris M (2006) Mt-DNA phylogeography of Lissotriton boscai (Caudata, Salamandridae): evidence for old, multiple refugia in an Iberian endemic. Mol Ecol 15:3375–3388PubMedCrossRefGoogle Scholar
  62. Mascort R (1998) Distribution and status of the European pond turtle, Emys orbicularis, in Catalonia. Mertensiella 10:177–186Google Scholar
  63. Moll D, Moll EO (2004) The ecology, exploitation, and conservation of river turtles. Oxford University Press, New YorkGoogle Scholar
  64. Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 76:5269–5273PubMedCrossRefGoogle Scholar
  65. Pinho C, Harris DJ, Ferrand N (2007) Contrasting patterns of population subdivision and historical demography in three western Mediterranean lizard species inferred from mitochondrial DNA variation. Mol Ecol 16:1191–1205PubMedCrossRefGoogle Scholar
  66. Pleguezuelos JM, Márquez R, Lizana M (2002) Atlas y Libro Rojo de los Anfibios y Reptiles de España. Dirección General de la Conservación de la Naturaleza-AHE, MadridGoogle Scholar
  67. Primack RB (1993) Essentials of conservation biology. Sinauer Associates, SunderlandGoogle Scholar
  68. Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959PubMedGoogle Scholar
  69. Recuero E, Iraola A, Rubio X, Machordom A, García-París M (2007) Mitochondrial differentiation and biogeography of Hyla meridionalis (Anura: Hylidae): an unusual phylogeographical pattern. J Biogeogr 34:1207–1219CrossRefGoogle Scholar
  70. Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225Google Scholar
  71. Rito de Araújo P, Segurado P, dos Santos NR (1997) Bases para a consevaçao das tartarugas de áuga doce (Emys orbicularis e Mauremys leprosa). Instituto da Conservaçao da Natureza, LisboaGoogle Scholar
  72. Roe JH, Georges A (2007) Heterogeneous wetland complexes, buffer zones, and travel corridors: Landscape management for freshwater reptiles. Biol Conserv 135:67–76CrossRefGoogle Scholar
  73. Rozas J, Rozas R (1997) DnaSP version 2.0: A novel software package for extensive molecular population genetics analysis. Comput Appl Biosci 13:307–311PubMedGoogle Scholar
  74. Schleich HH, Boehme W (1994) Kupferzeitliche Schildkrötenreste aus der Grabung von Durankulak bei Tolbuchin in NO-Bulgarien (Reptilia: Testudines: Testudo graeca, Emys orbicularis). Mitteilungen der Bayerischen Staatssammlung fuer Palaeontologie und Historische Geologie 34:199–211Google Scholar
  75. Schwartz T, Karl S (2005) Population and conservation genetics of the gopher tortoise (Gopherus polyphemus). Conserv Genet 6:917–928CrossRefGoogle Scholar
  76. Sites JW Jr, Fitzsimmons NN, da Silva NJ Jr, Cantarelli VH (1999) Conservation genetics of the Giant Amazon River Turtle (Podocnemis expansa): inferences from two classes of molecular markers. Chelonian Conserv Biol 3:454–463Google Scholar
  77. Slatkin M (1995) A measure of population subdivision based on microsatellite allele frequencies. Genetics 139:457–462PubMedGoogle Scholar
  78. Sokal RR, Rohlf FJ (1995) Biometry, 3rd edn. W.H. Freeman, New YorkGoogle Scholar
  79. Spinks PQ, Shaffer HB (2005) Range-wide molecular analysis of the western pond turtle (Emys marmorata): cryptic variation, isolation by distance, and their conservation implications. Mol Ecol 14:2047–2064PubMedCrossRefGoogle Scholar
  80. Spinks PQ, Shaffer HB, Iverson J, McCord W (2004) Phylogenetic hypotheses for the turtle family Geomydidae. Mol Phylogenet Evol 32:164–182PubMedCrossRefGoogle Scholar
  81. Starkey DE, Shaffer HB, Burke RL, Forstner MRJ, Iverson JB, Janzen FJ, Rhodin AGJ, Ultsch GR (2003) Molecular systematics, phylogeography, and the effects of Pleistocene glaciation in the painted turtle (Chrysemys picta) complex. Evolution 57:119–128PubMedGoogle Scholar
  82. Takezaki N, Nei M (1996) Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA. Genetics 144:389–399PubMedGoogle Scholar
  83. Templeton AR, Crandall KA, Sing CF (1992) A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequence data III. Cladogram estimation. Genetics 132:619–633PubMedGoogle Scholar
  84. van Dijk PP, Stuart BL, Rhodin AGJ (2000) Asian turtle trade: proceedings of a workshop on conservation and trade of freshwater turtles and tortoises in Asia. Chelonian Res Monogr 2:1–164Google Scholar
  85. Weir BS, Cockerham CC (1984) Estimating F-statistics for the analysis of population structure. Evolution 38:1358–1370CrossRefGoogle Scholar
  86. Wright S (1943) Isolation by distance. Genetics 28:114–138PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Guillermo Velo-Antón
    • 1
  • Mario García-París
    • 2
  • Adolfo Cordero Rivera
    • 1
  1. 1.Grupo de Ecoloxía Evolutiva, Departamento de Ecoloxía e Bioloxía AnimalUniversidade de VigoPontevedraSpain
  2. 2.Museo Nacional de Ciencias NaturalesMadridSpain

Personalised recommendations