A microsatellite-based method for genotyping diploid and triploid water frogs of the Rana esculenta hybrid complex

Authors

  • DITTE G. CHRISTIANSEN

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    1. Departments of Evolutionary and Population Biology, Biological Institute, University of Copenhagen, Universitetsparken 15, 2100 Copenhagen, Denmark
      Ditte G. Christiansen, Fax: 0045 3532 1250; E-mail: dgchristiansen@bi.ku.dk
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Ditte G. Christiansen, Fax: 0045 3532 1250; E-mail: dgchristiansen@bi.ku.dk

Abstract

Rana esculenta is a hybrid between Rana lessonae (LL) and Rana ridibunda (RR), and hybrids may be diploid (LR) or triploid (LLR or LRR). Genotypes can be roughly determined from erythrocyte size and morphometry in adult frogs, but accurate genotyping requires more labourious methods. Here I demonstrate that both the L and R genomes have specific microsatellite alleles, and that genotype and ploidy can be accurately inferred from the quantitative ratio of PCR-amplified (polymerase chain reaction-amplified) genome-specific alleles. This method greatly facilitates genotyping in DNA studies of the R. esculenta complex and allows analysis of badly preserved samples and embryos.

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