title: Novel genomic resources for a climate change sensitive mammal: characterization of the American pika transcriptome.
abstract: BACKGROUND
  When faced with climate change, species must either shift their home range or adapt in situ in order to maintain optimal physiological balance with their environment. The American pika (Ochotona princeps) is a small alpine mammal with limited dispersal capacity and low tolerance for thermal stress. As a result, pikas have become an important system for examining biotic responses to changing climatic conditions. Previous research using amplified fragment length polymorphisms (AFLPs) has revealed evidence for environmental-mediated selection in O. princeps populations distributed along elevation gradients, yet the anonymity of AFLP loci and lack of available genomic resources precluded the identification of associated gene regions. Here, we harnessed next-generation sequencing technology in order to characterize the American pika transcriptome and identify a large suite of single nucleotide polymorphisms (SNPs), which can be used to elucidate elevation- and site-specific patterns of sequence variation.
  
  
  RESULTS
  We constructed pooled cDNA libraries of O. princeps from high (1400 m) and low (300 m) elevation sites along a previously established transect in British Columbia. Transcriptome sequencing using the Roche 454 GS FLX titanium platform generated 780 million base pairs of data, which were assembled into 7,325 high coverage contigs. These contigs were used to identify 24,261 novel SNP loci. Using high resolution melt analysis, we developed 17 of these SNPs into genotyping assays, which were validated with independent DNA samples from British Columbia Canada and Oregon State USA. In addition, we detected haplotypes in the NADH dehydrogenase subunit 5 of the mitochondrial genome that were fixed and different among elevations, suggesting that this may be an informative target gene for studying the role of cellular respiration in local adaptation. We also identified contigs that were unique to each elevation, including a high elevation-specific contig that was a positive match with the hemoglobin alpha chain from the plateau pika, a species restricted to high elevation steppes in Asia. Elevation-specific contigs may represent candidate regions subject to differential levels of gene expression along this elevation gradient.
  
  
  CONCLUSIONS
  To our knowledge, this is the first broad-scale, transcriptome-level study conducted within the Ochotonidae, providing novel genomic resources for studying pika ecology, behaviour and population history.
authors.affiliation: Department of Biology, University of British Columbia, Okanagan Campus 3333 University Way, Kelowna, BC V1V 1V7, Canada. matt.lemay@ubc.ca
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Lemay
authors.forename: Philippe
authors.initials: P
authors.lastname: Henry
authors.forename: Clayton T
authors.initials: CT
authors.lastname: Lamb
authors.forename: Kelsey M
authors.initials: KM
authors.lastname: Robson
authors.forename: Michael A
authors.initials: MA
authors.lastname: Russello
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057231:Climate Change; D020451:Contig Mapping; D004272:DNA, Mitochondrial / Q000235:genetics; D004777:Environment; D020869:Gene Expression Profiling; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D006239:Haplotypes; D059014:High-Throughput Nucleotide Sequencing; D007793:Lagomorpha / Q000235:genetics*; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000737:chemistry / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D004272:DNA, Mitochondrial; D009245:NADH Dehydrogenase