Isolation and characterization of 21 polymorphic microsatellite loci in the iconic Australian lungfish, Neoceratodus for
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TECHNICAL NOTE
Isolation and characterization of 21 polymorphic microsatellite loci in the iconic Australian lungfish, Neoceratodus forsteri, using the Ion Torrent next-generation sequencing platform Joel A. Huey • Kathryn M. Real • Peter B. Mather • Vincent Chand • David T. Roberts • Thomas Espinoza • Andrew McDougall • Peter K. Kind Steven Brooks • Jane M. Hughes
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Received: 15 February 2013 / Accepted: 21 February 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We isolated and characterized 21 microsatellite loci in the vulnerable and iconic Australian lungfish, Neoceratodus forsteri. Loci were screened across eight individuals from the Burnett River and 40 individuals from the Pine River. Genetic diversity was low with between one and six alleles per locus within populations and a maximum expected heterozygosity of 0.774. These loci will now be available to assess effective population sizes and genetic structure in N. forsteri across its natural range in South East Queensland, Australia. Keywords Neoceratodus forsteri Lungfish Ion Torrent Microsatellite
The Australian Lungfish, Neoceratodus forsteri (Krefft, 1870) is one of five extant members of the ancient, air-breathing Dipnoi (lungfishes), and is the only species endemic to J. A. Huey (&) K. M. Real J. M. Hughes Australian Rivers Institute, Griffith University, Brisbane, QLD 4111, Australia e-mail: [email protected] P. B. Mather V. Chand Science and Engineering Faculty, Queensland University of Technology, Brisbane, QLD 4001, Australia D. T. Roberts Seqwater, Brisbane, QLD 4001, Australia T. Espinoza A. McDougall Queensland Department of Natural Resources and Mines, Bundaberg, QLD 4670, Australia P. K. Kind S. Brooks Queensland Department of Agriculture, Fisheries and Forestry, Brisbane, QLD 4001, Australia
Australia. Its range includes the Burnett, Mary, Brisbane, North Pine and Coomera catchments, all found on the east coast of the continent in southeast Queensland. Under the Australian Environment Protection and Biodiversity Conservation Act 1999, N. forsteri is listed as ‘‘vulnerable’’, due to losses or reductions in quality of breeding and nursery habitat and the ongoing water infrastructure development in the area (Arthington 2009; Pusey et al. 2004). Previous research has revealed low genetic variation and inbreeding for N. forsteri, using mitochondrial DNA sequence data and allozymes (Frentiu et al. 2001). However, to better estimate levels of variation and describe the spatial genetic structure of N. forsteri, more variable molecular markers are required. In addition to these questions, recently developed analyses in conservation genetics, such as the indirect estimation of effective population size (Ne) using molecular data (Luikart et al. 2010), requires more loci and more variable loci than have been previously available. As such, the development of microsatellite loci is a priority for the conservation of this species. Here, we describe the development of microsatellite markers, using the Ion Torrent pl
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