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Australian Journal of Zoology Australian Journal of Zoology Society
Evolutionary, molecular and comparative zoology
RESEARCH ARTICLE

Characterisation of microsatellites for Litoria nannotis (Amphibia : Hylidae), an endangered waterfall frog endemic to the Australian Wet Tropics

Robert Puschendorf A B E , Erica V. Todd C and Michael G. Gardner D
+ Author Affiliations
- Author Affiliations

A School of Biological and Marine Sciences, Plymouth University, Drake Circus, Plymouth, Devon PL4 8AA, UK.

B College of Marine and Environmental Sciences, James Cook University, Townsville, Qld 4811, Australia.

C Department of Anatomy, University of Otago, Dunedin 9016, New Zealand.

D School of Biological Sciences, Flinders University, GPO Box 2100, Adelaide, SA 5001, Australia, and Australian Centre for Evolutionary Biology and Biodiversity, University of Adelaide, Adelaide, SA 5005, Australia, and Evolutionary Biology Unit, South Australian Museum, North Terrace, Adelaide, SA 5000, Australia.

E Corresponding author. Email: robert.puschendorf@plymouth.ac.uk

Australian Journal of Zoology 64(6) 390-394 https://doi.org/10.1071/ZO16072
Submitted: 18 October 2016  Accepted: 2 March 2017   Published: 23 March 2017

Abstract

Litoria nannotis is an endangered waterfall frog from the wet tropics region in north Queensland that has suffered significant population declines due to the emerging fungal disease known as chytridiomycosis. The species has two deeply divergent lineages, and we used 454 shotgun sequencing of DNA extracted from one individual of the northern lineage to identify and design PCR primers for 576 microsatellite loci. Thirty markers were tested for amplification success and variability in a population sample from each lineage. Of these, 17 were found to be polymorphic in the northern lineage and 10 loci were polymorphic in the southern lineage. Numbers of alleles per locus ranged from 2 to 14 (mean = 6.47, s.d. = 4.02) for the northern lineage (17 polymorphic loci), and from 2 to 8 (mean = 5.40, s.d. = 2.55) in the southern lineage (10 polymorphic loci). Levels of heterozygosity were high in both lineages (northern mean HE = 0.63, s.d. = 0.21, range = 0.27–0.89; southern mean HE = 0.57, s.d. = 0.25, range = 0.18–0.81). These loci will be useful in understanding the genetic variation and connectivity amongst populations of this species recovering from mass population declines due to disease.

Additional keywords: 454 GSFLX, population declines, shotgun sequencing.


References

Bell R. C. MacKenzie J. B. Hickerson M. J. Chavarría K. L. Cunningham M Williams S Moritz C 2012 Comparative multi-locus phylogeography confirms multiple vicariance events in co-distributed rainforest frogs. Proceedings of the Royal Society B: Biological Sciences 279 991 999 10.1098/rspb.2011.1229

Benjamini, Y., and Hochberg, Y. (1995). Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society, Series B: Methodological 57, 289–300.

Berger, L., Speare, R., Daszak, P., Green, D. E., Cunningham, A. A., Goggin, C. L., Slocombe, R., Ragan, M. A., Hyatt, A. D., McDonald, K. R., Hines, H. B., Lips, K. R., Marantelli, G., and Parkes, H. (1998). Chytridiomycosis causes amphibian mortality associated with population declines in the rainforests of Australia and central America. Proceedings of the National Academy of Sciences of the United States of America 95, 9031–9036.
Chytridiomycosis causes amphibian mortality associated with population declines in the rainforests of Australia and central America.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXkvFaltbc%3D&md5=14ce13b2554e46c7df865a156184e323CAS |

Cunningham, M. (2002). Identification and evolution of Australian torrent treefrogs (Anura: Hylidae: Litoria nannotis group). Memoirs of the Queensland Museum 48, 93–102.

Gardner, M. G., Fitch, A. J., Bertozzi, T., and Lowe, A. J. (2011). Rise of the machines – recommendations for ecologists when using next generation sequencing for microsatellite development. Molecular Ecology Resources 11, 1093–1101.
Rise of the machines – recommendations for ecologists when using next generation sequencing for microsatellite development.Crossref | GoogleScholarGoogle Scholar |

Holleley, C. E., and Geerts, P. G. (2009). MULTIPLEX MANAGER 1.0: a cross-platform computer program that plans and optimizes multiplex PCR. BioTechniques 46, 511–517.
MULTIPLEX MANAGER 1.0: a cross-platform computer program that plans and optimizes multiplex PCR.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXns1ajsr4%3D&md5=777c4da2a58119693d713ae5f17698edCAS |

Kalinowski, S., Taper, M., and Marshall, T. (2007). Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology 16, 1099–1106.
Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment.Crossref | GoogleScholarGoogle Scholar |

Meglécz, E., Costedoat, C., Dubut, V., Gilles, A., Malausa, T., Pech, N., and Martin, J.-F. (2010). QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26, 403–404.
QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects.Crossref | GoogleScholarGoogle Scholar |

Meglécz, E., Nève, G., Biffin, E., and Gardner, M. G. (2012). Breakdown of phylogenetic signal: a survey of microsatellite densities in 454 shotgun sequences from 154 non model eukaryote species. PLoS One 7, e40861.
Breakdown of phylogenetic signal: a survey of microsatellite densities in 454 shotgun sequences from 154 non model eukaryote species.Crossref | GoogleScholarGoogle Scholar |

Peakall, R., and Smouse, P. E. (2012). GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research – an update. Bioinformatics 28, 2537–2539.
GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research – an update.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhsVehtbjI&md5=f36d0bcb5d48b2df5606e724dad5896fCAS |

Puschendorf, R., Hoskin, C. J., Cashins, S. D., McDonald, K., Skerratt, L. F., Vanderwal, J., and Alford, R. A. (2011). Environmental refuge from disease-driven amphibian extinction. Conservation Biology 25, 956–964.
Environmental refuge from disease-driven amphibian extinction.Crossref | GoogleScholarGoogle Scholar |

Raymond, M., and Rousset, F. (1995). GENEPOP (version 1.2): Population genetics software for exact tests and ecumenicism. The Journal of Heredity 86, 248–249.
GENEPOP (version 1.2): Population genetics software for exact tests and ecumenicism.Crossref | GoogleScholarGoogle Scholar |

Richards, S. J., McDonald, K. R., and Alford, R. A. (1994). Declines in populations of Australia’s endemic tropical rainforest frogs. Pacific Conservation Biology 1, 66–77.
Declines in populations of Australia’s endemic tropical rainforest frogs.Crossref | GoogleScholarGoogle Scholar |

Rousset, F. (2008). Genepop’007: a complete reimplementation of the Genepop software for Windows and Linux. Molecular Ecology Resources 8, 103–106.
Genepop’007: a complete reimplementation of the Genepop software for Windows and Linux.Crossref | GoogleScholarGoogle Scholar |

Rozen, S., and Skaletsky, H. J. (2000). Primer3 on the WWW for general users and for biologist programmers. In ‘Bioinformatics Methods and Protocols: Methods in Molecular Biology’. (Eds S. Krawetz, and S. Misener.) pp. 365–386. (Humana Press: Totowa, NJ.)

Schneider, C. J., Cunningham, M., and Moritz, C. (1998). Comparative phylogeography and the history of endemic vertebrates in the Wet Tropics rainforests of Australia. Molecular Ecology 7, 487–498.
Comparative phylogeography and the history of endemic vertebrates in the Wet Tropics rainforests of Australia.Crossref | GoogleScholarGoogle Scholar |

Van Oosterhout, C., Hutchinson, B., Wills, D., and Shipley, P. (2004). MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Molecular Ecology Notes 4, 535–538.
MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXnvFOktb8%3D&md5=9cd3b93c23d1169be30f5281343cfa4eCAS |