Kozlov’s pika is a rare and endangered lagomorph species with a limited distribution in the southern Kunlun Mountains in western China. Because of its endangered status, Kozlov’s pika is considered a priority species for research and conservation action. Genetic variation and molecular evolution of the Kozlov’s pika were studied based on a total of 14 individuals from four locations along the eastern boundary between Xinjiang and Tibet province (35.20–36.48°N, 86.08–83.04°E) on extremely high elevation (usually over 4800 m a.s.l.). The density of local populations was about 3–4 per ha, living in a typical alpine desert grassland habitat. The complete mitochondrial cytochrome b (cytb) gene was amplified and sequenced. Based on the cytb gene sequences the genetic variation and molecular evolution were analyzed. Unexpected high haplotype diversity (0.956 ± 0.045) but low nucleotide diversity (0.00537 ± 0.00126) was found, indicating past demographic expansion. Significant partitioning of variance (P <0.01) among populations (46.7%), and within populations (53.3%), indicating low level of genetic differentiations among local populations. Our results gave an optimistic survival status of Kozlov’s pika at the genetic level. Bayes Empirical Bayes analysis with model M2a and M8 detected three positively selected amino acid sites at the significance level of 0.05. The mutant types with either or both of the mutations aspartic acid to asparagine and glutamic acid to lysine had higher isoelectric point values. We suggested these mutant types might have biological significance to help individuals to adapt to the extremely high elevation habitats.
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