Molecular Ecology Notes (2005) 5, 205–207 doi: 10.1111/j.1471-8286.2005.00877.x © 2005 Blackwell Publishing Ltd Blackwell Publishing, Ltd. PRIMER NOTE Characterization of microsatellite loci in the endemic mound spring snail Fonscochlea accepta and cross species amplification in four other hydrobiid snails JESSICA WORTHINGTON WILMER,* JANE M. HUGHES,† JING MA† and CHRIS WILCOX‡ *Biodiversity Program, Queensland Museum, PO Box 3300, Brisbane QLD 4101, Australia, †Australian School of Environmental Studies, Griffith University, Nathan QLD 4111, Australia, ‡ Department of Zoology and Entomology, University of Queensland, Brisbane QLD 4072, Australia Abstract We identified and characterized nine microsatellite primer pairs from Fonscochlea accepta, a species of hydrobiid snail endemic to artesian mound springs associated with the Great Artesian Basin in arid South Australia. The loci were highly polymorphic, with five to 30 alleles per locus. Gene diversity, estimated as expected heterozygosity ranged from 0.364 to 0.851, and was generally matched by levels of observed heterozygosity (0.373 – 0.829). Cross- species amplification trials with four other hydrobiid species associated with these south- ern mound springs showed that these primers will be useful for genetic analyses of these other endemic snails. Keywords: artesian mound springs, Hydrobiidae, metapopulations, microsatellites Received 23 September 2004; revision accepted 10 November 2004 The mound springs ecosystem of the Great Artesian Basin (GAB) in northern South Australia is of national cultural, economic and biological significance. These artesian springs form an integral part of the history of many of the Aboriginal groups in the region and defined the primary route for expansion of European colonists into the Australian interior. Economically, the GAB covers 22% of outback and regional Australia and is the region’s primary water resource. Production from the basin is currently valued at $A 3.2 billion (GABCC 1998). This is set to increase as higher-valued industries take over water use from pastoralism. The springs support rare and delicate flora and fauna, of which a significant number of species are now endangered because of over extraction of artesian water for agricultural, industrial and municipal uses. As a result, the unique flora and fauna of these systems have recently been listed as an ‘endangered community’ under the Australian Environmental Protection & Biodiversity Conservation Act of 1999. The springs are known to contain over 40 species of indigenous aquatic and amphibious snails of the family, Hydrobiidae, with some species restricted to a single spring (Ponder et al . 1989). There is increasing evidence that the aquatic invertebrates, including the snails, operate as classical metapopulations (Ponder et al . 1989; WMC 2002). In order to obtain a greater understanding of connectivity among springs and metapopulation dynamics of these snails, we developed primers for nuclear microsatellite loci and tested these primers on four other hydrobiid snails of the genera Fonscochlea and Trochidrobia which inhabit the springs of South Australia. DNA for library construction was isolated by homogen- ization of entire snails ( c . 3 mm in diameter) using a DNeasy Tissue Kit (QIAGEN). Approximately 10 μ g of genomic DNA was digested for 3 h with restriction enzyme Sau 3A1. After separation on a 1.5% agarose gel, DNA fragments in the size range of 200 – 800 base pairs were excised, purified and ligated to an equal volume of plasmid vector pUC18 (Amersham Pharmacia). The plasmids had previously been digested with Bam HI and dephosphorylated to create overhanging ends to match those resulting from the Sau 3A1 digest. Recombinant plasmids were electroporated into competent Escherichia coli cells (strain JM109, Promega) and incubated for an hour at 37 ° C. Cells were spread on to agar plates containing LB-Ampicillin and incubated over- night at 37 ° C to promote selective growth of transformed colonies. A total of 2200 recombinant colonies were picked from plates and incubated overnight in a grid formation on Correspondence: Jessica Worthington Wilmer, Fax: (61-7) 3846 1226; E-mail: jessicaww@qm.qld.gov.au