RESEARCH ARTICLE


Development of New Microsatellite (STR) Markers for Montagu’s Harrier (Circus pygargus) via 454 Shot-Gun Pyrosequencing



Susann Janowski1, *, Markus A. Grohme2, Marcus Frohme2, Michael Wink1
1 Institute of Pharmacy and Molecular Biotechnology, Department Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany
2 Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences, Hochschulring 1, 15745 Wildau, Germany


© 2014 Janowski et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Institute of Pharmacy and Molecular Biotechnology, Department Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany; Tel: 0049-6221-54-4847; Fax: 0049-6221-54-4884; E-mail: Janowski@uni-heidelberg.de


Abstract

During the last decades the ground-breeding Montagu’s harrier (Circus pygargus, Linnaeus, 1758) has changed its breeding habitats in Europe to agricultural areas in which many local populations would be close to extinction without a special nest protection regime. Although Montagu’s harrier is a well-studied species in terms of ecology and breeding biology, its genetic structure and population genetics are almost unknown. As there is a lack of good genetic markers we developed a set of 19 microsatellite markers comprising 16 new STR markers which were identified by next-generation sequencing (NGS) using 454 shot-gun pyrosequencing of genomic DNA. The STR markers were arranged into three multiplex PCR sets for high throughput genotyping and characterised. The marker set provides a powerful tool for kinship analysis. The combined non-exclusion probability for parent pairs was 1.13*10-11. Only three loci showed PIC values < 0.50. In total, 121 known family relationships were compared with genetically calculated ones to test the markers suitability for parentage analysis. In 97.5% of all cases full-sibships were accurately determined and 97.6% of all mothers were assigned correctly to their chicks. The present multiplex PCR panels can be used to investigate several hypotheses concerning breeding behaviour, kinship, exchange rates between populations and phylogeography.

Keywords: Circus pygargus, Dipetalogaster maxima, genotyping, microsatellites, Montagu’s harrier, multiplex PCR, next-generation sequencing, parentage analysis.