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Rapid molecular sexing of three-spined sticklebacks, Gasterosteus aculeatus L., based on large Y-chromosomal insertions

  • Animal Genetics • Original Paper
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Abstract

There is a need for rapid and reliable molecular sexing of three-spined sticklebacks, Gasterosteus aculeatus, the supermodel species for evolutionary biology. A DNA region at the 5′ end of the sex-linked microsatellite Gac4202 was sequenced for the X chromosome of six females and the Y chromosome of five males from three populations. The Y chromosome contained two large insertions, which did not recombine with the phenotype of sex in a cross of 322 individuals. Genetic variation (SNPs and indels) within the insertions was smaller than on flanking DNA sequences. Three molecular PCR-based sex tests were developed, in which the first, the second or both insertions were covered. In five European populations (from DE, CH, NL, GB) of three-spined sticklebacks, tests with both insertions combined showed two clearly separated bands on agarose minigels in males and one band in females. The tests with the separate insertions gave similar results. Thus, the new molecular sexing method gave rapid and reliable results for sexing three-spined sticklebacks and is an improvement and/or alternative to existing methods.

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Acknowledgements

We are grateful to Gerald Heckel and Marion Mehlis for discussions. We thank Dagmar Wenzel for practical assistance. Dominique Mazzi is acknowledged for breeding sticklebacks. Thomas Schaper and Jürgen Wittler gave permission to catch sticklebacks from the Euskirchen population. Peter Snelderwaart and George Wintermans are gratefully thanked for providing us with Dutch sticklebacks, and Ricarda Modarressie for the Scottish samples. We thank an anonymous reviewer for improving the quality of the manuscript. TCMB and CRL thank the Swiss National Science Foundation for financial support.

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Author notes

  1. Joachim G. Frommen

    Present address: Department of Behavioural Ecology, Institute of Ecology and Evolution, University of Bern, Wohlenstrasse 50a, 3032, Hinterkappelen, Switzerland

  2. Carlo R. Largiadèr

    Present address: University Institute of Clinical Chemistry, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

Authors and Affiliations

  1. Institute for Evolutionary Biology and Ecology, University of Bonn, An der Immenburg 1, 53121, Bonn, Germany

    Theo C. M. Bakker & Joachim G. Frommen

  2. Computational and Molecular Population Genetics Lab (CMPG), Abteilung Populationsgenetik, Zoologisches Institut, Universität Bern, Baltzerstrasse 6, 3012, Bern, Switzerland

    Thomas Giger & Carlo R. Largiadèr

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  1. Theo C. M. Bakker
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  2. Thomas Giger
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  3. Joachim G. Frommen
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  4. Carlo R. Largiadèr
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Contributions

TCMB and CRL designed the research; TCMB and CRL obtained funding for the study; TG and JGF conducted the experimental work; TCMB, TG and CRL analysed the data; TCMB wrote the manuscript and was supported by CRL and JGF.

Corresponding author

Correspondence to Theo C. M. Bakker.

Ethics declarations

The study conforms to the Association for the Study of Animal Behaviour guidelines for the use of animals in research as well as to the legal requirements of Switzerland and Germany.

Funding

This study was funded by a grant from the Swiss National Science Foundation to TCMB and CRL (SNF grant no. 31-52276.97).

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by: Maciej Szydlowski

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Bakker, T.C.M., Giger, T., Frommen, J.G. et al. Rapid molecular sexing of three-spined sticklebacks, Gasterosteus aculeatus L., based on large Y-chromosomal insertions. J Appl Genetics 58, 401–407 (2017). https://doi.org/10.1007/s13353-017-0399-0

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  • DOI: https://doi.org/10.1007/s13353-017-0399-0

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