Advertisement

Isolation, characterization and cross-species amplification of new microsatellite markers for three opossum species of the Didelphidae family

  • Isabela M. G. Dias
  • George Amato
  • Heitor M. Cunha
  • Rob DeSalle
  • Adriano P. Paglia
  • Jennifer K. Peterson
  • Cleusa G. Fonseca
Technical Note

Abstract

The American opossums from the Didelphidae family (Order: Didelphimorphia) are on the IUCN Red List, and their threatened status varies from extinct to data deficient. Although microsatellites have already been developed for some species, only a small amount of population genetic analysis of the didelphid wild populations has been made using them. In this work, we developed new microsatellites and tested for cross-species amplification for one species from the subfamily Caluromyinae, Caluromys philander, and two from the subfamily Didelphinae, Didelphis aurita and Metachirus nudicaudatus, to determine the degree of gene flow and isolation of populations of those species in some fragments of the Atlantic Forest in Minas Gerais state, Brazil. Among all microsatellite loci amplified, 18 from C. philander, 9 from D. aurita, and 17 from M. nudicaudatus were polymorphic. Cross-species amplification of these polymorphic microsatellite loci were performed in these three species and in one additional related species Micoureus paraguayanus. Our data showed that microsatellites were not highly conserved among Didelphidae species.

Keywords

Didelphidae Microsatellite Caluromys Didelphis Metachirus Micoureus Opossum 

Notes

Acknowledgments

We would like to thank our colleagues from the Sackler Institute of Comparative Genomics, especially Matt Leslie; colleagues from the Mammalogy Lab of the General Biology Department of ICB-UFMG, who helped us with the field work and who furnished samples, especially Raquel Moura. This study is funded by the CNPq Project PELD-PERD, the CAPES Foundation (Ph.D. student fellowship), and the Sackler Institute for Comparative Genomics—AMNH.

References

  1. Brownstein M, Carpenter J, Smith J (1996) Modulation of nontemplated nucleotide addition by Taq polymerase: primer modifications that facilitate genotyping. BioTechniques 20:1004–1010PubMedGoogle Scholar
  2. Dias IMG, Amato G, Carvalho MRS, Cunha HM, Paglia AP, DeSalle R, Fonseca CG (2008) Characterization of eight microsatellite loci in the woolly mouse opossum, Micoureus paraguayanus isolated from Micoureus demerarae. Mol Ecol Resour 8:345–347CrossRefGoogle Scholar
  3. Faircloth BC, Shamblin B, Eo SH et al (2008) Microsatellites from the Virginia opossum (Didelphis virginiana). In: NCBI Entrez Nucleotide. http://www.ncbi.nlm.nih.gov/sites/entrez of subordinate document. Accessed Aug 12, 2009
  4. Gardner LA (2007) Mammals of South America volume 1: Marsupials xenarthrans, shrews, and bats. The University of Chicago Press, Chicago, ILGoogle Scholar
  5. Glenn TC, Schable NA (2005) Isolating microsatellite DNA loci. Methods Enzymol 395:202–222CrossRefPubMedGoogle Scholar
  6. Guillemin ML, Lavergne A, Catzeflis F (2000) Microsatellite markers in the common grey four-eyed opossum (Philander opossum Didelphidae, Marsupialia). Mol Ecol 9(9):1440–1442CrossRefPubMedGoogle Scholar
  7. IUCN (2008) 2008 IUCN Red list of threatened species. http://www.iucnredlist.org. Accessed Aug 12, 2009
  8. Kalinowski ST, Taper ML, Marshall TC (2007) Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Mol Ecol 16:1006–1099CrossRefGoogle Scholar
  9. Lavergne A, Douady C, Catzeflis FM (1999) Isolation and characterization or microsatellite loci in Didelphis marsupialis (Marsupialia: Didelphidae). Mol Ecol 8(3):517–518PubMedGoogle Scholar
  10. Rodrigues FP, Rocha FS, Garcia JE et al (2006) Isolation and characterization of microsatellite loci in the woolly mouse opossum, Micoureus paraguayanus (Marsupialia : Didelphimorphia). Mol Ecol Notes 6:686–688CrossRefGoogle Scholar
  11. Rousset F (2008) Genepop’007: a complete re-implementation of the genepop software for Windows and Linux. Mol Ecol Resour 8(1):103–106CrossRefGoogle Scholar
  12. Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols. Humana Press, Totowa, NJ, pp 365–386Google Scholar
  13. Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. CSHL Press, New YorkGoogle Scholar
  14. Samollow PB, Kammerer CM, Mahaney SM et al (2004) First-generation linkage map of the gray, short-tailed opossum, Monodelphis domestica, reveals genome-wide reduction in female recombination rates. Genetics 166(1):307–329CrossRefPubMedGoogle Scholar
  15. Samollow PB, Gouin N, Miethke P et al (2007) A microsatellite-based, physically anchored linkage map for the gray, short-tailed Opossum (Monodelphis domestica). Chromosome Res 15:269–281PubMedGoogle Scholar
  16. Shuelke M (2000) An economic method for fluorescent labeling of PCR fragments. Nat Biotechnol 18:233–234CrossRefGoogle Scholar
  17. Temnykh S, DeClerck G, Lukashova A (2001) Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): frequency, length variation, transposon associations, and genetic marker potential. Genome Res 11:1441–1452CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Isabela M. G. Dias
    • 1
    • 2
  • George Amato
    • 2
  • Heitor M. Cunha
    • 1
  • Rob DeSalle
    • 2
  • Adriano P. Paglia
    • 3
  • Jennifer K. Peterson
    • 2
  • Cleusa G. Fonseca
    • 1
  1. 1.Departamento de Biologia Geral, ICBUniversidade Federal de Minas GeraisBelo HorizonteBrazil
  2. 2.Sackler Institute for Comparative Genomics, American Museum of Natural HistoryNew YorkUSA
  3. 3.Conservation International BrazilBelo HorizonteBrazil

Personalised recommendations