Development of polymorphic microsatellite markers for the microendemic pupfishes Cyprinodon julimes and C. pachycephalus
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TECHNICAL NOTE
Development of polymorphic microsatellite markers for the microendemic pupfishes Cyprinodon julimes and C. pachycephalus Evan W. Carson • Rochelle R. Beasley • Kenneth L. Jones • Stacey L. Lance • Ma de Lourdes Lozano-Vilano • Lilia Vela-Valladares • Iris Banda-Villanueva Thomas F. Turner • Mauricio De la Maza-Benignos
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Received: 21 March 2013 / Accepted: 26 March 2013 / Published online: 2 April 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We developed microsatellite loci for the Julimes pupfish, Cyprinodon julimes. Twenty-five loci were screened across 19 individuals from Julimes Spring, Chihuahua, Mexico. The number of alleles per locus ranged from 2 to 14, observed heterozygosity ranged from 0.105 to 0.947, and the probability of identity values ranged from 0.022 to 0.588. We then tested for cross-amplification in the bighead pupfish, C. pachycephalus; twenty-three individuals from San Diego de Alcala´, Chihuahua, Mexico, were screened across the 20 loci that amplified cleanly. These new loci will be used for long-term genetic monitoring of these critically endangered species. Keywords Cyprinodon Illumina Microsatellite PAL_FINDER PCR primers SSR E. W. Carson (&) T. F. Turner Department of Biology and Museum of Southwestern Biology, University of New Mexico, Albuquerque, NM 87131, USA e-mail: [email protected] R. R. Beasley S. L. Lance Savannah River Ecology Laboratory, University of Georgia, Aiken, SC 29802, USA K. L. Jones Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045, USA M. de Lourdes Lozano-Vilano Laboratorio de Ictiologı´a, Facultad de Ciencias Biolo´gicas, Universidad Auto´noma de Nuevo Leo´n, 66450 San Nicola´s de los Garza, NL, Mexico L. Vela-Valladares I. Banda-Villanueva M. De la Maza-Benignos Pronatura Noreste, A.C, Loma Larga 235, Col. Loma Larga, 64710 Monterrey, NL, Mexico
The pupfishes Cyprinodon julimes and C. pachycephalus are microendemic pupfishes of the Chihuahuan Desert, Mexico. These species are limited, respectively, to the small desert spring systems at Julimes and San Diego de Alcala´, Chihuahua, where they exhibit the highest tolerances to extended thermal stress known for teleosts (Minckley and Minckley 1986; De la Maza-Benignos et al. 2009). Due to their microgeographic distributions and physiological specialization, these pupfishes are highly vulnerable to extinction, especially from human exploitation of water resources in this arid region. Both species are listed as federally endangered in Mexico (SEMARNAT 2010), with C. pachycephalus also listed as critically endangered in the IUCN Red List of Threatened Species (IUCN 2012). In accordance with conservation agreements between Pronatura Noreste A. C. and landowners at Julimes and San Diego de Alcala´, newly developed microsatellite loci reported here will be used for long-term genetic monitoring under the ‘‘Vital Signs’’ program for these critically imperiled species (De la Maza-Benignos et al. 2012). Total D
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