TECHNICAL NOTE Isolation and characterization of microsatellite loci in the plain-winged woodcreeper Dendrocincla turdina (Aves, Dendrocolaptidae) Ana Cristina Fazza • Pedro M. Galetti Jr. Received: 26 April 2010 / Accepted: 21 May 2010 / Published online: 18 June 2010 Ó Springer Science+Business Media B.V. 2010 Abstract Dendrocincla turdina has a poor ability to survive in fragmented areas and inhabits the depleted Brazilian Atlantic Forest. For the assessment of genetic diversity in this species, nineteen microsatellite loci were isolated and characterized, 11 of which were polymorphic for the population studied. Observed and expected het- erozigosity ranged from 0.07 to 0.80 and 0.08 to 0.91, respectively. Cross-amplification was successfully obtained with two other species from the family Dendrocolaptidae: Xiphorhynchus fuscus and Sittasomus griseicapillus. The newly developed primers reported here constitute a useful tool for genetic population analyses on D. turdina and, potentially, other related species. Keywords Genetic conservation Á Passeriformes Á Atlantic forest Á Population genetics The plain-winged woodcreeper (Dendrocincla turdina) is an insectivorous species (Aves: Dendrocolaptidae) that inhabits the understory of the Brazilian Atlantic Forest (Sick 1997). Woodcreepers are highly vulnerable to envi- ronmental changes (Aleixo and Vielliard 1995; Poletto et al. 2004). Population declines and even extinction in depleted areas are related to habitat specialization (Marantz et al. 2003). Dendrocincla turdina undergoes a decline in abundance in fragmented areas. The species is already critically endangered in the state of Rio Grande do Sul (southern Brazil) (Marques et al. 2002) and locally extinct in the southeastern portion of the state of Minas Gerais (south- eastern Brazil) (Ribon et al. 2003). This sensitivity underscores the importance of genetic conservation studies for the species. The aim of the present study was to isolate and characterize microsatellite loci for D. turdina, as there are none described in the literature. Such information could successfully be used to assess the genetic diversity of the species. A microsatellite-enriched library was obtained using a protocol adapted from Hamilton et al. (1999). Genomic DNA isolated from blood samples was digested with RsaI (GE Healthcare) and the DNA fragments were ligated to double strand SNX linkers (Hamilton et al. 1999). The products were enriched with eight biotinylated tetranucle- otide probes. After hybridization with these probes, DNA fragments containing microsatellites were captured using magnetic beads (Streptavidin Magnesphere Paramagnetic Particles, Promega). Enriched DNA was cloned using the pGem-T Easy Vector (Promega) and 96 clones were sequenced in an ABI3730XL automated sequencer. The sequences were analyzed using the CID program (Freitas et al. 2008) for microsatellite identification and design of the primers. Nineteen microsatellites were identified and their primers were constructed using a 5 0 18 base pair M13 universal sequence strategy (Schuelke 2000) for indirect fluoro-labeling detection. The loci were amplified in 10 ll PCR containing 19 amplification buffer, 0.25 mM of each dNTP, 0.8 pmol of fluorescently-labeled M13 primer (NED or 6-FAM), 0.2 pmol of the primer containing the M13 sequence, 0.8 pmol of the other primer, 1.5 mM of MgCl 2 , 5 ng of template DNA (50 ng/ll) and 0.5 U of Taq DNA Polymerase (Invitrogen). The reaction cycling parameters were 94°C (5 min), 30 cycles of 94°C (30 s), primer-pair specific annealing temperature (Table 1) for A. C. Fazza Á P. M. Galetti Jr. (&) Departamento de Gene ´tica e Evoluc ¸a ˜o, Universidade Federal de Sa ˜o Carlos, Via Washington Luis km 235, Caixa Postal 676, Sa ˜o Carlos, SP 13565-905, Brazil e-mail: galettip@ufscar.br 123 Conservation Genet Resour (2011) 3:5–7 DOI 10.1007/s12686-010-9258-6