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.
This is a preview of subscription content, log in via an institution to check access.
References
Brownstein M, Carpenter J, Smith J (1996) Modulation of nontemplated nucleotide addition by Taq polymerase: primer modifications that facilitate genotyping. BioTechniques 20:1004–1010
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 8:345–347
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
Gardner LA (2007) Mammals of South America volume 1: Marsupials xenarthrans, shrews, and bats. The University of Chicago Press, Chicago, IL
Glenn TC, Schable NA (2005) Isolating microsatellite DNA loci. Methods Enzymol 395:202–222
Guillemin ML, Lavergne A, Catzeflis F (2000) Microsatellite markers in the common grey four-eyed opossum (Philander opossum Didelphidae, Marsupialia). Mol Ecol 9(9):1440–1442
IUCN (2008) 2008 IUCN Red list of threatened species. http://www.iucnredlist.org. Accessed Aug 12, 2009
Kalinowski ST, Taper ML, Marshall TC (2007) Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Mol Ecol 16:1006–1099
Lavergne A, Douady C, Catzeflis FM (1999) Isolation and characterization or microsatellite loci in Didelphis marsupialis (Marsupialia: Didelphidae). Mol Ecol 8(3):517–518
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 6:686–688
Rousset F (2008) Genepop’007: a complete re-implementation of the genepop software for Windows and Linux. Mol Ecol Resour 8(1):103–106
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 365–386
Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. CSHL Press, New York
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):307–329
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 15:269–281
Shuelke M (2000) An economic method for fluorescent labeling of PCR fragments. Nat Biotechnol 18:233–234
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 11:1441–1452
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Dias, I.M.G., Amato, G., Cunha, H.M. et al. Isolation, characterization and cross-species amplification of new microsatellite markers for three opossum species of the Didelphidae family. Conservation Genet Resour 1, 405 (2009). https://doi.org/10.1007/s12686-009-9094-8
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s12686-009-9094-8