Testing and Utility of Microsatellite loci in Hyla orientalis

Ergül Kalaycı T., Özdemir N.

2nd International Symposium on Biodiversity Research, Rize, Turkey, 18 - 20 November 2020, pp.208

  • Publication Type: Conference Paper / Summary Text
  • City: Rize
  • Country: Turkey
  • Page Numbers: pp.208
  • Karadeniz Technical University Affiliated: No


Microsatellites are one of the most widely used tools in population genetics and conservation biology. These markers have been developed for some amphibian species. Assessment of appropriate genetic markers for population genetic studies in amphibians is critical because amphibians are declining globally. Microsatellite loci are widely used in population and conservation genetic studies of amphibians. We tested amplification success and polymorphism in 19 (WHA1-20, WHA1-25, WHA1-103, WHA1-67, WHA5-22A, WHA1-104, WHA1-61, Ha-A11, HaB12, HaB5R3, Ha-A119, Ha-A119, Ha-A136, Ha-A110, Ha-A139, Ha-A116, Ha-T60, Ha-T66, Ha-T67 and Ha-T68) previously developed microsatellite loci, in Hyla orientalis species from Rize, Turkey. Eight individuals were used in this study. DNA was isolated according to the manufacturer’s instruction. The PCR reactions were carried out in 25 μL reaction mixture, containing 12.5 μL master mix, 2.5 μL primer mix, 1 μL of DNA template and remaining concentrations was filled with RNase-free water. Forward primer fluorescently labeled with FAM, VIC, NED or PET. We succeed to amplify twelve microsatellite loci, but one of (WHA1-104) them were not genotyped in fragment analysis. According to microsatellite analysis, 66.67 % of the amplifying loci were polymorphic. The mean number of alleles, effective alleles, observed heterozygosity (HO) and expected heterozygosity (HE) were found as 2.92, 2.15, 0.39 and 0.37, respectively. Most of those loci were used in H. orientalis by other studies in the literature. Some of them were amplified in previous studies, but some of them were amplificated for the first time for H. orientalis in this study (like Ha-A11, Ha-136 and HaB5R3). Our results provide additional genetic support for the existing literature. It is important to evaluate and reveal useful microsatellite loci in target species for future conservation studies, aiming to address taxonomic uncertainties, or to study genetic variability and differentiation among populations.

Keywords: Amphibia, Microsatellite, Hyla orientalis