Mikroszatellita-markerkészlet fejlesztése parlagi sasok (Aquila heliaca) egyedi azonosításához rokon fajokban leírt markerek segítségével

Abstract

Background: DNA profiling allows the study of population structure, dispersal and survival of protected species and can play an important role in mitigating wildlife crime. The eastern imperial eagle (Aquila heliaca) is a globally vulnerable raptor which has been actively monitored in Hungary for decades, using DNA profiling based on microsatellite markers. However, due to the rapid growth of the population, additional markers are required for reliable individual identification in the future. Objectives: Our aim was to increase the resolution of the microsatellite marker set currently used for the individual identification of the eastern imperial eagle with cross-species markers described in related species. Materials and Methods: We tested 26 cross-species microsatellite markers, out of which six were described in the Spanish imperial eagle (A. adalberti), 13 in the Japanese golden eagle (A. chrysaetos japonica) and seven in the white-tailed eagle (Haliaeetus albicilla). First, we tested the markers in monoplex PCR reactions in 4–15 unrelated individuals, using DNA from shed feathers of breeding individuals collected between 2011 and 2022. Then, we created multiplexes of the polymorphic markers where possible to reduce genotyping costs. Lastly, we tested the markers for Hardy-Weinberg equilibrium and the presence of null alleles, and estimated the probabilities of identity for the new marker set. Results and Discussion: In general, the polymorphism and amplification success of markers were positively correlated with relatedness between the eastern imperial eagle and the source species. We selected 17 markers for the final marker set, seven previously used and ten newly tested markers (two eastern imperial eagle, ten Spanish imperial eagle, three Japanese golden eagle and two white-tailed eagle markers). All markers were in Hardy-Weinberg equilibrium and no null alleles were detected. The probability of identity decreased compared to the old marker set (from 10 -6 to 10 -14), ensuring the reliable individual identification of eastern imperial eagles in the future.