The unique genetic adaptation of the Himalayan wolf to high-altitudes and consequences for conservation

The Himalayan wolf seems uniquely adapted to life at high-altitudes of the Himalayas and the Tibetan Plateau. Through a non-invasive survey we confirm the presence of the Himalayan wolf across the Nepalese Himalayas and its phylogenetic distinctness based on mitochondrial and nuclear DNA. We use the data generated from 287 scat and hair samples combined with a reference dataset including canid samples from around the globe. The Himalayan wolf forms a genetically distinct lineage based on 1) 242bp of D-loop and 508bp of cytochrome b (mtDNA), 2) the ZF gene of both sex chromosomes, 3) a microsatellite panel of 17 nuclear loci, and 4) four non-synonymous SNPs in four hypoxia pathway related (functional) nuclear genes. The SNP analysis indicates a genetic adaptation to cope with the hypoxic stresses in the high altitude habitats which we did not find in the Holarctic grey wolf. Based on analysis of divergence time from full mitochondrial genomes we estimate that the Himalayan wolf diverged from the Holarctic grey wolf complex 691,000–740,000 years before the present day. We provide first insights into the population status of the Himalayan wolf in Nepal with nuclear genotyping revealing counts of 12, 16, and 2 wolf individuals in the three study areas Humla (384 km2), Dolpa (1,088 km2), and Kanchenjunga Conservation Area (368 km2) respectively. The methods presented here offer a complete toolkit for the non-invasive monitoring of this wolf lineage. Nepal holds a significant population of this unique wolf across its Himalayan landscapes and we recommend the country takes a leading role on its protection. Keywords: Canis (lupus) himalayensis, High-altitude adaptation, Himalayan wolf, Nepal, Non-invasive sampling, Phylogen