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Int. J. Mol. Sci. 2012, 13(10), 12259-12267; doi:10.3390/ijms131012259
Article

Development of 22 Polymorphic Microsatellite Loci for the Critically Endangered Morato’s Digger Toad, Proceratophrys moratoi

1,* , 2
, 3
 and 1
1 Department of Structural and Functional Biology, Institute of Biology, University of Campinas-UNICAMP, Campinas, SP, CEP 13083-863, Brazil 2 Department of Zoology, Institute of Biosciences, State University of São Paulo-UNESP, Botucatu, SP, CEP 18618-970, Brazil 3 Center for Molecular Biology and Genetic Engineering, Institute of Biology, University of Campinas-NICAMP, Campinas, SP, CEP 13083-875, Brazil
* Author to whom correspondence should be addressed.
Received: 22 August 2012 / Revised: 29 August 2012 / Accepted: 3 September 2012 / Published: 25 September 2012
(This article belongs to the Section Bioinorganic Chemistry)
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Abstract

The Morato’s digger toad (Proceratophrys moratoi) inhabits Brazilian moist savannas and is critically endangered due to its very limited geographic distribution, reduced number of isolated populations, and evidence of population decline and local extinctions. With the objective of providing tools for the genetic study of the species, 22 polymorphic microsatellite loci were isolated and screened using DNA extracted from samples of oral mucosa cells obtained from 113 individuals representing five remnant P. moratoi populations in the Brazilian state of São Paulo. These markers presented 2–18 alleles per locus, polymorphism information content (PIC) of 0.02–0.87, observed heterozygosity of 0.02–0.96 and expected heterozygosity of 0.02–0.87. Three of the loci deviated significantly from Hardy–Weinberg equilibrium in one of the populations, possibly due to the presence of null alleles. Significant linkage disequilibrium was also detected between three pairs of loci. The molecular markers developed in this study were able to discriminate each of the individuals sampled (identity analysis). This means that they will be extremely useful for future genetic studies applied to the conservation of P. moratoi, providing a baseline for estimating the levels of genetic diversity, pedigrees, inbreeding, and population structure, which will be essential for the development of effective genetic management programs.
Keywords: Proceratophrys moratoi; endangered species; microsatellite; population genetics; Morato’s digger toad; conservation genetics Proceratophrys moratoi; endangered species; microsatellite; population genetics; Morato’s digger toad; conservation genetics
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Arruda, M.P.; Costa, W.P.; Silva, C.C.; Pimentel, S.M.R. Development of 22 Polymorphic Microsatellite Loci for the Critically Endangered Morato’s Digger Toad, Proceratophrys moratoi. Int. J. Mol. Sci. 2012, 13, 12259-12267.

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