The development of 10 novel polymorphic microsatel

Authors: Adam D Miller Anthony Van Rooyen Oisín F Sweeney Nick S Whiterod Andrew R Weeks

Publish Date: 2013/05/04

Volume: 40, Issue: 7, Pages: 4415-4419

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Abstract

The Glenelg spiny crayfish Euastacus bispinosus is an iconic freshwater invertebrate of south eastern Australia and listed as ‘endangered’ under the Environment Protection and Biodiversity Conservation Act 1999 and ‘vulnerable’ under the International Union for Conservation of Nature’s Red List The species has suffered major population declines as a result of overfishing low environmental flows the introduction of invasive fish species and habitat degradation In order to develop an effective conservation strategy patterns of gene flow genetic structure and genetic diversity across the species distribution need to be clearly understood In this study we develop a suite of polymorphic microsatellite markers by next generation sequencing A total of 15 polymorphic loci were identified and 10 characterized using 22 individuals from the lower Glenelg River We observed low to moderate genetic variation across most loci mean number of alleles per locus = 280 mean expected heterozygosity = 036 with no evidence of individual loci deviating significantly from Hardy–Weinberg equilibrium Marker independence was confirmed with tests for linkage disequilibrium and analyses indicated no evidence of null alleles across loci Individuals from two additional sites Crawford River Victoria Ewens Ponds Conservation Park South Australia were genotyped at all 10 loci and a preliminary investigation of genetic diversity and population structure was undertaken Analyses indicate high levels of genetic differentiation among sample locations F ST  = 049 while the Ewens Ponds population is genetically homogeneous indicating a likely small founder group and ongoing inbreeding Management actions will be needed to restore genetic diversity in this and possibly other at risk populations These markers will provide a valuable resource for future population genetic assessments so that an effective framework can be developed for implementing conservation strategies for E bispinosusThe fieldwork for this project was partially funded by the Norman Wettenhall Foundation and grants from the federal governments Caring for Our Country program awarded to the Friends of Mt Gambier Area Parks Thanks to Dave Mossop Lauren Kivisalu and volunteers from the Friends of Mount Gambier Area Parks and the wider community for assistance whilst collecting samples All sample collection was conducted under DENR Wildlife Research permit E259631 PIRSA SA Fisheries Exemption permit 9902414 Victorian Fisheries Act 1995 permit RP1092 Victorian Natural Parks Act 1975 and Flora and Flora Guarantee Act 1988 permit 10006431 and Victorian DPI Animal Ethics permit 2112 Thanks to the Department of Environment Water and Natural Resources South Australia the South East Water Conservation and Drainage Board Parks Victoria and several private landholders in South East South Australia for access to sites

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