Development of primers for amplification and further research on the D-loop control region for the Common Spadefoot toad, Pelobates fuscus (Amphibia)

作者：Vanbrabant Jeroen^*; Goddeeris Boudewijn; Martens Koen; Schon Isa

来源：Conservation Genetics Resources, 2009, 1(1): 495-499.

DOI：10.1007/s12686-009-9115-7

• Pelobates fuscus
• Common Spadefoot toad
• Mitochondrial DNA
• D-loop control region
• Genetic diversity
• Amphibian conservation genetics

Pelobates fuscus is an endangered toad occurring throughout Europe, for which suitable genetic markers are still rare. Here, we developed universal PCR primers for the amplification of the mitochondrial D-loop control region by long range PCR, which will be suitable for P. fuscus and other Pelobatidae. Sequence analyses of the 1,578 bp of the control region from two Flemish populations in Zonhoven and Peer ( Belgium) indicated that this mitochondrial region is sufficiently variable to separate populations at a small geographic scale. Based on the obtained long D-loop sequences, additional, specific primers were developed for standard PCR protocols, to amplify a shorter part of 857 bp containing most of the genetic variability. Our results indicate that the D-loop region is also for Amphibians a suitable locus for conservation genetic purposes, as it allows estimating genetic diversity in recently diverged populations from their mitochondrial genomes.

• 出版日期2009-12