Data from: Parallel molecular evolution in pathways, genes, and sites in high-elevation hummingbirds revealed by comparative transcriptomics

High-elevation organisms experience shared environmental challenges that include low oxygen availability, cold temperatures, and intense UV radiation. Consequently, repeated evolution of the same genetic mechanisms may occur across high-elevation taxa. To test this prediction, we investigated the extent to which the same biochemical pathways, genes, or sites were subject to parallel molecular evolution for 12 Andean hummingbird species (family: Trochilidae) representing several independent transitions to high elevation across the phylogeny. Across high-elevation species, we discovered parallel evolution for several pathways and genes with evidence of positive selection. In particular, positively selected genes were frequently part of cellular respiration, metabolism, or cell death pathways. To further examine the role of elevation in our analyses, we compared results for low- and high-elevation species and tested different thresholds for defining elevation categories. In analyses with different elevation thresholds, positively selected genes reflected similar functions and pathways, even though there were almost no specific genes in common. For example, EPAS1 (HIF2), which has been implicated in high-elevation adaptation in other vertebrates, shows a signature of positive selection when high-elevation is defined broadly (> 1500 m), but not when defined narrowly (> 2500 m). While a few biochemical pathways and genes change predictably as part of hummingbird adaptation to high-elevation conditions, independent lineages have rarely adapted via the same substitution

mlim_transcriptome_scripts

Bioinformatics and data analysis scripts used in the study. The purpose of each script is briefly described in the readme file

MinION-Based DNA Barcoding of Preserved and Non-Invasively Collected Wildlife Samples

The ability to sequence a variety of wildlife samples with portable, field-friendly equipment will have significant impacts on wildlife conservation and health applications. However, the only currently available field-friendly DNA sequencer, the MinION by Oxford Nanopore Technologies, has a high error rate compared to standard laboratory-based sequencing platforms and has not been systematically validated for DNA barcoding accuracy for preserved and non-invasively collected tissue samples. We tested whether various wildlife sample types, field-friendly methods, and our clustering-based bioinformatics pipeline, SAIGA, can be used to generate consistent and accurate consensus sequences for species identification. Here, we systematically evaluate variation in cytochrome b sequences amplified from scat, hair, feather, fresh frozen liver, and formalin-fixed paraffin-embedded (FFPE) liver. Each sample was processed by three DNA extraction protocols. For all sample types tested, the MinION consensus sequences matched the Sanger references with 99.29%–100% sequence similarity, even for samples that were difficult to amplify, such as scat and FFPE tissue extracted with Chelex resin. Sequencing errors occurred primarily in homopolymer regions, as identified in previous MinION studies. We demonstrate that it is possible to generate accurate DNA barcode sequences from preserved and non-invasively collected wildlife samples using portable MinION sequencing, creating more opportunities to apply portable sequencing technology for species identification

Climate change refugia and habitat connectivity promote species persistence

Background: Climate change refugia, areas buffered from climate change relative to their surroundings, are of increasing interest as natural resource managers seek to prioritize climate adaptation actions. However, evidence that refugia buffer the effects of anthropogenic climate change is largely missing. Methods: Focusing on the climate-sensitive Belding\u27s ground squirrel (Urocitellus beldingi), we predicted that highly connected Sierra Nevada meadows that had warmed less or shown less precipitation change over the last century would have greater population persistence, as measured by short-term occupancy, fewer extirpations over the twentieth century, and long-term persistence measured through genetic diversity. Results: Across California, U. beldingi were more likely to persist over the last century in meadows with high connectivity that were defined as refugial based on a suite of temperature and precipitation factors. In Yosemite National Park, highly connected refugial meadows were more likely to be occupied by U. beldingi. More broadly, populations inhabiting Sierra Nevada meadows with colder mean winter temperatures had higher values of allelic richness at microsatellite loci, consistent with higher population persistence in temperature-buffered sites. Furthermore, both allelic richness and gene flow were higher in meadows that had higher landscape connectivity, indicating the importance of metapopulation processes. Conversely, anthropogenic refugia, sites where populations appeared to persist due to food or water supplementation, had lower connectivity, genetic diversity, and gene flow, and thus might act as ecological traps. This study provides evidence that validates the climate change refugia concept in a contemporary context and illustrates how to integrate field observations and genetic analyses to test the effectiveness of climate change refugia and connectivity. Conclusions: Climate change refugia will be important for conserving populations as well as genetic diversity and evolutionary potential. Our study shows that in-depth modeling paired with rigorous fieldwork can identify functioning climate change refugia for conservation