Characterization of novel microsatellite loci for Hetaerina americana damselflies, and cross-amplification in other spec
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TECHNICAL NOTE
Characterization of novel microsatellite loci for Hetaerina americana damselflies, and cross-amplification in other species Christopher N. Anderson • Gregory F. Grether
Received: 28 July 2012 / Accepted: 22 August 2012 Ó Springer Science+Business Media B.V. 2012
Abstract Hetaerina damselflies are distributed throughout the neotropics. We developed eleven microsatellite loci for the damselfly Hetaerina americana. Microsatellites were tested for polymorphism on a panel of 24 individuals. The number of alleles ranged from 2 to 6, observed heterozygosity from 0.080 to 0.701, and the fixation index from -0.266 to 1.000. Cross-amplification was tested in 7 different species in the genus Hetaerina from the United States and Mexico. These microsatellite loci will be useful for studies of population structure and gene flow in H. americana. Keywords americana
Microsatellite Damselfly Hetaerina
Hetaerina is a neotropical damselfly genus with 37 species (Garrison 1990). H. americana is among the most broadly distributed representative of the genus, ranging from Canada to northern Central Americana and is among the most highly studied with regards to its behavioral ecology and immunocompetence (e.g. Grether 1996; ContrerasGardun˜o et al. 2006). Although Hetaerina americana is widespread in North America, several populations are vulnerable (NatureServe 2012). Furthermore, we aim to use
C. N. Anderson (&) Departamento de Ecologı´a Evolutiva, Instituto de Ecologı´a, Universidad Nacional Auto´noma de Me´xico, Circuito Exterior s/n, Apdo., Postal 70-275, 04510 Mexico, D.F., Mexico e-mail: [email protected] G. F. Grether Department of Ecology and Evolutionary Biology, University of California, Los Angeles, 621 Charles E. Young Drive South, Los Angeles, CA 90095-1606, USA
these microsatellite loci to characterize the population genetics of several species in the genus, including those that have imperiled national or regional statuses (e.g. H. titia, NatureServe 2012). Here we describe the isolation and characterization of 11 novel microsatellite loci from H. americana as a population genetics characterization resource. Individuals were obtained from riparian sites near Castroville, Texas, United States (29.343°N 98.882°W); Arroyo de Piedra, Veracruz, Mexico (19.456°N 96.482°W); and Armeria, Colima, Mexico (18.961°N 103.965°W). Insects were captured with aerial nets, stored in 95 % ethanol, and then refrigerated at -80 °C until analysis. DNA was extracted from wing muscle using the Qiagen QIAamp DNA Mini Kit using the manufacturer’s protocol. 500 ng of DNA was prepared for whole genome shotgun sequencing on the Roche Genome Sequencer FLX instrument using the GS FLX Titanium Rapid Library Preparation Kit (Roche Applied Sciences, Indianapolis, USA) following the manufacturer’s protocol. The library was quantified for DNA fragment size distribution and concentration (Agilent 2100 Bioanalyzer) and then processed with the GS FLX emulsion polymerase chain reaction (PCR) and sequencing kits. Sequencing was per
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