Eur. J. Entomol. 109 (1): 129-134, 2012 | 10.14411/eje.2012.016

Development of novel microsatellite markers for a specialist species of Lepidoptera, Boloria aquilonaris (Nymphalidae), based on 454 sequences

Sofie VANDEWOESTIJNE*,1, Camille TURLURE1,2, Michel BAGUETTE2,3
1 Biodiversity Research Centre, Earth and Life Institute, Université Catholique de Louvain (UCL), Croix du Sud 4, B-1348 Louvain-la-Neuve, Belgium
2 CNRS UMR 7204, Muséum National d'Histoire Naturelle, Dept Ecologie & Gestion de la Biodiversité, 55 rue Buffon, 75005 Paris, France
3 CNRS USR 2936, Station d'Ecologie Expérimentale, 09200 Moulis, France

Microsatellites are the most common markers used in population and conservation genetic studies. However, their isolation is laborious and expensive. In some taxa, such as Lepidoptera, it is particularly difficult to isolate microsatellite markers due to the high similarity of the flanking regions of different loci and the presence of null alleles. Here we isolated microsatellites of the endangered butterfly Boloria aquilonaris using 454 GS-FLX Titanium pyro-sequences of biotin enriched DNA libraries and tested the success of cross-amplification on the sister-species B. eunomia. Fifteen polymorphic microsatellite loci were isolated in B. aquilonaris using initially 101 stringently designed primer pairs. Unlike in many other studies of microsatellite isolation in Lepidoptera, few null alleles were detected and only at very low frequencies. Additionally, the raw data set can still be used for the isolation of other microsatellite loci. None of the selected polymorphic loci for B. aquilonaris gave clear banding patterns for B. eunomia, although about 15 other loci gave promising banding patterns for the latter species. Low intra- and inter-specific transferability of developed markers in this study also lends support to the hypothesis that the evolution of the genome of Lepidoptera is dissimilar from that of other organisms.

Keywords: Lepidoptera, Nymphalidae, microsatellite, pyrosequencing, cross-amplification

Received: March 17, 2011; Accepted: August 22, 2011; Published: January 3, 2012

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References

  1. ABDELKRIM J., ROBERTSON B.C., STANTON J.A.L. & GEMMELL N.J. 2009: Fast, cost-effective development of species-specific microsatellite markers by genomic sequencing. Biotechniques 46: 185-191 Go to original source...
  2. ALTSCHUL S.F., MADDEN T.L., SCHAFFER A.A., ZHANG J., ZHANG Z., MILLER W. & LIPMAN D.J. 1997: Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389-3402 Go to original source...
  3. BAGUETTE M. 2003: Long distance dispersal and landscape occupancy in a metapopulation of the cranberry fritillary butterfly. Ecography 26: 153-160 Go to original source...
  4. BAGUETTE M., GOFFART P. & DE BAST B. 1992: Modification de la distribution et du statut des Lepidopteres Rhopaloceres en Belgique depuis 1900. Mem. Soc. R. Belge Entomol. 35: 591-596
  5. BENJAMINI Y. & HOCHBERG Y. 1995: Controlling the false discovery rate - a practical and powerful approach to multiple testing. J. R. Statist. Soc. (B, Methodol.) 57: 289-300
  6. CAMPBELL D., DUCHESNE P., BERNATCHEZ L. 2003: AFLP utility for population assignment studies: analytical investigation and empirical comparison with microsatellites. Mol. Ecol. 12: 1979-1991 Go to original source...
  7. CHAPUIS M.P. & ESTOUP A. 2007: Microsatellite null alleles and estimation of population differentiation. Mol. Biol. Evol. 24: 621-631 Go to original source...
  8. CHEN M.H. & DORN S. 2010: Cross-amplification of microsatellites from the codling moth Cydia pomonella to three other species of the tribe Grapholitini (Lepidoptera: Tortricidae). Mol. Ecol. Res. 10: 1034-1037 Go to original source...
  9. DAKIN E.E. & AVISE J.C. 2004: Microsatellite null alleles in parentage analysis. Heredity 93: 504-509 Go to original source...
  10. HABEL J.C., FINGER A., MEYER M., SCHMITT T. & ASSMANN T. 2008: Polymorphic microsatellite loci in the endangered butterfly Lycaena helle (Lepidoptera: Lycaenidae). Eur. J. Entomol. 105: 361-362 Go to original source...
  11. HARPER G.L., PIYAPATTANAKORN S., GOULSON D. & MACLEAN N. 2000: Isolation of microsatellite markers from the Adonis blue butterfly (Lysandra bellargus). Mol. Ecol. 9: 1948-1949 Go to original source...
  12. JARNE P. & LAGODA P.J.L. 1996: Microsatellites, from molecules to populations and back. Trends Ecol. Evol. 11: 424-429 Go to original source...
  13. KIJAS J.M.H., FOWLER J.C.S., GARBETT C.A. & THOMAS M.R. 1994: Enrichment of microsatellites from the citrus genome using biotinylated oligonucleotide sequences bound to streptavidin-coated magnetic particles. Biotechniques 16: 656-662
  14. MALAUSA T., GILLES A., MEGLECZ E. ET AL. 2011: Highthroughput microsatellite isolation through 454 GS-FLX Titanium pyrosequencing of enriched DNA libraries. Mol. Ecol. Res. 11: 638-644 Go to original source...
  15. MEGLECZ E., PETENIAN F., DANCHIN E., D'ACIER A.C., RASPLUS J.Y. & FAURE E. 2004: High similarity between flanking regions of different microsatellites detected within each of two species of Lepidoptera: Parnassius apollo and Euphydryas aurinia. Mol. Ecol. 13: 1693-1700 Go to original source...
  16. MEGLECZ E., COSTEDOAT C., DUBUT V., GILLES A., MALAUSA T., PECH1 N. & MARTIN J.-F. 2010: QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26: 403-404 Go to original source...
  17. MEUDT H.M. & CLARKE A.C. 2007: Almost forgotten or latest practice? AFLP applications, analyses and advances. Trends Plant Sci. 12: 106-117 Go to original source...
  18. MIKHEYEV A.S., VO T., WEE B., SINGER M.C. & PARMESAN C. 2010: Rapid microsatellite isolation from a butterfly by de novo transcriptome sequencing: performance and a comparison with AFLP-derived distances. Plos One 5: e11212 Go to original source...
  19. MILLER C.R. & WAITS L.P. 2003: The history of effective population size and genetic diversity in the Yellowstone grizzly (Ursus arctos): Implications for conservation. Proc. Natn. Acad. Sci. U. S. A. 100: 4334-4339 Go to original source...
  20. MOUSSON L., NEVE G. & BAGUETTE M. 1999: Metapopulation structure and conservation of the cranberry fritillary Boloria aquilonaris in (Lepidoptera, Nymphalidae) in Belgium. Biol. Conserv. 87: 285-293 Go to original source...
  21. NEVE G. & MEGLECZ E. 2000: Microsatellite frequencies in different taxa. Trends Ecol. Evol. 15: 376-377 Go to original source...
  22. NEW T.R. 1997: Are Lepidoptera an effective "umbrella group" for biodiversity conservation? J. Insect Conserv. 1: 5-12 Go to original source...
  23. PEMBERTON J.M., SLATE J., BANCROFT D.R. & BARRETTA J.A. 1995: Nonamplifying alleles at microsatellite loci: a caution for parentage and population studies. Mol. Ecol. 4: 249-252 Go to original source...
  24. ROUSSET F. 2008: GENEPOP '007: a complete re-implementation of the GENEPOP software for Windows and Linux. Mol. Ecol. Res. 8: 103-106 Go to original source...
  25. SARHAN A. 2006: Isolation and characterization of five microsatellite loci in the Glanville fritillary butterfly (Melitaea cinxia). Mol. Ecol. Notes 6: 163-164 Go to original source...
  26. SHINDE D., LAI Y.L., SUN F.Z. & ARNHEIM N. 2003: Taq DNA polymerase slippage mutation rates measured by PCR and quasi-likelihood analysis: (CA/GT)(n) and (A/T)(n) microsatellites. Nucl. Acids Res. 31: 974-980 Go to original source...
  27. SINAMA M., DUBUT V., COSTEDOAT C., GILLES A., JUNKER M., MALAUSA T., MARTIN J.F., NEVE G., PECH N., SCHMITT T., ZIMMERMANN M. & MEGLECZ E. 2011: Challenges of microsatellite development in Lepidoptera: Euphydryas aurinia (Nymphalidae) as a case study. Eur. J. Entomol. 108: 261-266 Go to original source...
  28. TURLURE C., CHOUTT J., VAN DYCK H., BAGUETTE M. & SCHTICKZELLE N. 2010a: Functional habitat area as a reliable proxy for population size: case study using two butterfly species of conservation concern. J. Insect Conserv. 14: 379-388. Go to original source...
  29. TURLURE C., CHOUTT J., BAGUETTE M. & VAN DYCK H. 2010b: Microclimatic buffering and resource-based habitat in a glacial relict butterfly: significance for conservation under climate change. Global Change Biol. 16: 1883-1893 Go to original source...
  30. VALLONE P.M. & BUTLER J.M. 2004: AutoDimer: a screening tool for primer-dimer and hairpin structures. Biotechniques 37: 226-231
  31. VAN OOSTERHOUT C., HUTCHINSON W.F., WILLS D.P.M. & SHIPLEY P. 2004: MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol. Ecol. Notes 4: 535-538 Go to original source...
  32. VAN SWAAY C., CUTTELOD A., COLLINS S., MAES D., LOPEZ MUNGUIRA M., SASIC M., SETTELE J., VEROVNIK R., VERSTRAEL T., WARREN M., WIEMERS M. & WYNHOFF I. 2010: European Red List of Butterflies. Publications Office of the European Union, Luxembourg, xii + 44 pp
  33. VANDEWOESTIJNE S. & BAGUETTE M. 2002: The genetic structure of endangered populations in the Cranberry Fritillary, Boloria aquilonaris (Lepidoptera, Nymphalidae): RAPDs vs allozymes. Heredity 89: 439-445 Go to original source...
  34. VANDEWOESTIJNE S. & BAGUETTE M. 2004: Demographic versus genetic dispersal measures. Popul. Ecol. 46: 281-285 Go to original source...
  35. WARREN M.S., HILL J.K., THOMAS J.A., ASHER J., FOX R., HUNTLEY B., ROY D.B., TELFER M.G., JEFFCOATE S., HARDING P., JEFFCOATE G., WILLIS S.G., GREATOREX-DAVIES J.N., MOSS D. & THOMAS C.D. 2001: Rapid responses of British butterflies to opposing forces of climate and habitat change. Nature 414: 65-69 Go to original source...
  36. WATTIER R., ENGEL C.R., SAUMITOU-LAPRADE P. & VALERO M. 1998: Short allele dominance as a source of heterozygote deficiency at microsatellite loci: experimental evidence at the dinucleotide locus Gv1CT in Gracilaria gracilis (Rhodophyta). Mol. Ecol. 7: 1569-1573 Go to original source...
  37. WILLIAMS J.G.K., KUBELIK A.R., LIVAK J., RAFALSKI J.A. & TINGEY S.V. 1990: DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Res. 18: 6531-6535 Go to original source...