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Development of sexing primers in Glyptemys insculpta and Apalone spinifera turtles uncovers an XX/XY sex-determining system in the critically-endangered bog turtle Glyptemys muhlenbergii

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Abstract

In species or developmental stages where the sex of an individual cannot be reliably identified through external morphology, molecular markers can provide a critical tool to study sex-specific traits that are elusive otherwise. Here we generated two sets of sex-diagnostic PCR primers for each of two focal turtle species with contrasting genotypic sex determination (GSD) systems: the wood turtle, Glyptemys insculpta (XX/XY), and the spiny softshell turtle, Apalone spinifera (ZZ/ZW). These markers identified males and females with 100% accuracy as validated with numerous individuals of known sex. Notably, one of the markers developed for G. insculpta permitted the successful diagnosis of individual sex in the critically-endangered bog turtle, Glyptemys muhlenbergii, also with 100% accuracy. This cross-species application provided the first evidence that G. muhlenbergii shares an XX/XY sex-determining mechanism with G. insculpta, a finding with important evolutionary and conservation implications. Similarly, the markers from A. spinifera were successful in identifying the sex of two individuals (one male and one female) of the Chinese softshell turtle, Pelodiscus sinensis (ZZ/ZW). These cross-species observations highlight the potential applicability of these types of markers on closely related taxa that share a sex-determining mechanism, which should be tested in a case-by-case basis.

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Acknowledgements

This work was funded in part by a Bern W. Tryon Bog Fund Grant from the Knoxville Zoological Gardens to NV and RB, and National Science Foundation grant MCB 1244355 to NV. We greatly appreciate the assistance of Mike Ogle at Zoo Knoxville, Amanda Guthrie at the Virginia Zoo, and Steve DeSimone at Cold Spring Harbor Fish Hatchery and Aquarium for their assistance in this Project.

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Authors and Affiliations

  1. Department of Ecology, Evolution, and Organismal Biology, Iowa State University, Ames, USA

    Robert Literman, Srihari Radhakrishnan & Nicole Valenzuela

  2. Department of Biology, University of Northern Iowa, Cedar Falls, USA

    Jeff Tamplin

  3. Department of Biology, Hofstra University, Hempstead, USA

    Russell Burke

  4. Department of Ecology and Evolutionary Biology, University of Tennessee Knoxville, Knoxville, USA

    Cassie Dresser

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  1. Robert Literman
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Correspondence to Robert Literman.

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Literman, R., Radhakrishnan, S., Tamplin, J. et al. Development of sexing primers in Glyptemys insculpta and Apalone spinifera turtles uncovers an XX/XY sex-determining system in the critically-endangered bog turtle Glyptemys muhlenbergii . Conservation Genet Resour 9, 651–658 (2017). https://doi.org/10.1007/s12686-017-0711-7

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