, Volume 140, Issue 4–6, pp 159–167 | Cite as

Molecular evidence for the subspecific differentiation of blue sheep (Pseudois nayaur) and polyphyletic origin of dwarf blue sheep (Pseudois schaeferi)

  • Shuai Tan
  • Dandan Zou
  • Lei Tang
  • Gaochao Wang
  • Quekun Peng
  • Bo Zeng
  • Chen Zhang
  • Fangdong ZouEmail author


Blue sheep (Pseudois nayaur), a Central Asian ungulate with restricted geographic distribution, exhibits unclear variation in morphology and phylogeographic structure. The composition of species and subspecies in the genus Pseudois is controversial, particularly with respect to the taxonomic designation of geographically restricted populations. Here, 26 specimens including 5 dwarf blue sheep (Pseudois schaeferi), which were collected from a broad geographic region in China, were analyzed for 2 mitochondrial DNA fragments (cytochrome b and control region sequences). In a pattern consistent with geographically defined subspecies, we found three deeply divergent mitochondrial lineages restricted to different geographic regions. The currently designated two subspecies of blue sheep, Pseudois nayaur nayaur and Pseudois nayaur szechuanensis, were recognized in the phylogenetic trees. In addition, the Helan Mountain population showed distinct genetic characteristics from other geographic populations, and thus should be classified as a new subspecies. In contrast, dwarf blue sheep clustered closely with some blue sheep from Sichuan Province in the phylogenetic trees. Therefore, dwarf blue sheep appear to be a subset of Pseudois nayaur szechuanensis. After considering both population genetic information and molecular clock analysis, we obtained some relevant molecular phylogeographic information concerning the historical biogeography of blue sheep. These results also indicate that western Sichuan was a potential refugium for blue sheep during the Quaternary period.


Blue sheep Cytochrome b Mitochondrial DNA control region Phylogeographic structure 



We are grateful to Xining Zoo and Chongqing Zoo for providing samples. This research was funded by the Program for New Century Excellent Talents in University (NCET-10-06-07), National Natural Science Foundation of China (30970335), Scientific Research Foundation for Returned Scholars, Ministry of Education of China (20091001-9-9) and Supporting Program for the Leaders of Distinguished Young Scholars Subject of Sichuan Province (09ZQ026-044).


  1. Allen GM (1939) Zoological results of the second Dolan expedition to western China and eastern Tibet, 1934–1936. Proc Acad Nat Sci Phila 90:261–293Google Scholar
  2. Amadon D (1949) The seventy-five percent rule for subspecies. Condor 51:250–258CrossRefGoogle Scholar
  3. An Z, John EK, Warren LP, Stephen CP (2001) Evolution of Asian monsoons and phased uplift of the Himalaya-Tibetan plateau since Late Miocene times. Nature 411:62–66CrossRefGoogle Scholar
  4. Avise JC (1998) Phylogeography: the history and formation of species. Harvard University Press, HarvardGoogle Scholar
  5. Cai CP, Hu JC, Peng JT (1990) The dwarf blue sheep of western Sichuan. J East China Norm Univ (Mamm Ecol Suppl):90–95Google Scholar
  6. Cao LR, Wang XM, Fang SG (2003) A molecular phylogeny of bharal and dwarf blue sheep based on mitochondrial cytochrome b gene sequences. Acta Zool Sin 49:198–204Google Scholar
  7. Cao LR, Wang XM, Rao G, Zhang KJ, Wan QH, Fang SG (2004) Population genetic structure of blue sheep based on molecular evidence from mtDNA. Acta Zootax Sin 49:198–204Google Scholar
  8. Drummond AJ, Rambaut A (2007) BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evol Biol 7:214PubMedCrossRefGoogle Scholar
  9. Eythorsdottir E, Tapio M (2001) Origin and genetic diversity of north European sheep breeds. In: Eythorsdottir E et al (ed) Book of abstracts of the 52nd annual meeting of the European Association for Animal Production, Budapest, 26–29 Aug 2001, p 262Google Scholar
  10. Fang XM, Zhao ZJ et al (2005) Magnetostratigraphy of the Late Cenozoic Laojunmiao anticline in the northern Qilian Mountains and its implications for the northern Tibetan Plateau up lift. Sci China (Ser D) 48(7):1040–1051CrossRefGoogle Scholar
  11. Feng J, Lajia C, Taylor DJ, Webster MS (2001) Genetic distinctiveness of endangered dwarf blue sheep (Pseudois nayaur schaeferi): evidence from mitochondrial control region and Y-linked ZFY intron sequences. J Hered 92(1):9–15PubMedCrossRefGoogle Scholar
  12. Fu YX (1997) Statistical test of neutrality of mutations against population growth, hitchhiking and background selection. Genetics 147:915–925PubMedGoogle Scholar
  13. Groves CP (1978) The taxonomic status of the dwarf blue sheep (Artiodactyla: Bovidae). Saugetierk Mitt 26:177–183Google Scholar
  14. Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704PubMedCrossRefGoogle Scholar
  15. Harris RB (2007) Wildlife conservation in China: preserving the habitat of China’s wild west. M E Sharpe Inc Armonk, New YorkGoogle Scholar
  16. Harris RB (2008) Pseudois nayaur. In: IUCN 2011. IUCN Red List of Threatened Species. Version Jan 2011
  17. Hassanin A, Ropiquet A, Couloux A, Cruaud C (2009) Evolution of the mitochondrial genome in mammals living at high altitude: new insights from a study of the tribe Caprini (Bovidae, Antilopinae). J Mol Evol 68:293–310PubMedCrossRefGoogle Scholar
  18. Hewitt GM (1996) Some genetic consequences of ice ages, and their role in divergence and speciation. Biol J Linn Soc 58:247–276Google Scholar
  19. Hewitt G (2000) The genetic legacy of the Quaternary ice ages. Nature 405:907–913PubMedCrossRefGoogle Scholar
  20. Higgins D, Thompson J, Gibson T, Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  21. Huelsenback JP, Ronquist F (2001) MR-BAYES: Bayesian inference of phylogeny. Bioinformatics 17:754–755CrossRefGoogle Scholar
  22. Hull JM, Girman DJ (2005) Effects of Holocene climate change on the historical demography of migrating sharp-shinned hawks (Accipiter striatus velox) in North America. Mol Ecol 14:159–170PubMedCrossRefGoogle Scholar
  23. Ibrahim KM, Nichols RA, Hewitt GM (1996) Spatial patterns of genetic variation generated by different forms of dispersal during range expansion. Heredity 77:282–291CrossRefGoogle Scholar
  24. Irwin DM, Kocher TD, Wilson AC (1991) Evolution of the cytochrome-b gene of mammals. J Mol Evol 32:128–144PubMedCrossRefGoogle Scholar
  25. Jing M, Yu H, Wu S, Wang W, Zheng X (2007) Phylogenetic relationships in genus Niviventer (Rodentia: Muridae) in China inferred from complete mitochondrial cytochrome b gene. Mol Phylogenet Evol 44:521–529PubMedCrossRefGoogle Scholar
  26. Li J, Li B, Wang F (1981) The process of uplift of the Qinghai-Xizang Plateau. Geological and ecological studies of Qinghai-Xizang Plateau. Science Press, BeijingGoogle Scholar
  27. Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452PubMedCrossRefGoogle Scholar
  28. Luikart G, Gielly L, Excoffier L et al (2001) Multiple maternal origins and weak phylogeographic structure in domestic goats. Proc Natl Acad Sci USA 98:5927–5932PubMedCrossRefGoogle Scholar
  29. Mayr E (1963) Animal species and evolution. Belknap Press, CambridgeGoogle Scholar
  30. Posada D (2008) JModelTest: phylogenetic model averaging. Mol Biol Evol 25:1253–1256PubMedCrossRefGoogle Scholar
  31. Posada D, Buckley TR (2004) Model selection and model averaging in phylogenetics: advantages of Akaike Information Criterion and Bayesian approaches over likelihood ratio tests. Syst Biol 53:793–808PubMedCrossRefGoogle Scholar
  32. Rambaut A, Drummond AJ (2007) Tracer v1.4. Accessed 20 June 2008
  33. Sbisà E, Tanzariello F, Reyes A, Pesole G, Saccone C (1997) Mammalian mitochondrial D-loop region structural analysis: identification of new conserved sequences and their functional and evolutionary implications. Gene 205:125–140PubMedCrossRefGoogle Scholar
  34. Schafer E (1937) On the dwarf bharal (Pseudois spec. nov) and the large bharal (P. nahhor Hodgson) of the Tibet. Zool Garten Leipzig 9:263–278Google Scholar
  35. Schaller GB (1998) Wildlife of the Tibetan steppe. University of Chicago Press, ChicagoGoogle Scholar
  36. Shi Y, Zheng B, Li S (1990) Last glaciation and maximum glaciation in Qinghai-Xizang (Tibet) plateau. J Glaciol Geocryol 12:1–15Google Scholar
  37. Shi Y, Li J, Li B (1998) Uplift and environmental changes of Qinghai-Tibetan plateau in the Late Cenozoic. Guangdong Science and Technology Press, GuangzhouGoogle Scholar
  38. Tajima F (1989) Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123:585–595PubMedGoogle Scholar
  39. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  40. Wang X, Hoffmann RS (1987) Pseudois nayaur and Pseudois schaeferi. Mamm Spec 278:1–6CrossRefGoogle Scholar
  41. Wang Y, Wang XM (2000) Population ecology of dwarf blue sheep (Pseudois schaeferi). Biodivers Sci 11(1):59–62Google Scholar
  42. Wang XM, Peng JT, Zhou HM (2000) Preliminary observations on the distribution and status of dwarf blue sheep Pseudois schaeferi. Orxy 34:21–26Google Scholar
  43. Wang LM, Gu HJ, Luo LZ, Deng XX, Yan QG, Qin MH (2006a) Death cause of Pseudois nayaur at Rangtang County in Sichuan Province. J Prev Med Inf 22:392–394Google Scholar
  44. Wang XM, Cao LR, Liu ZS, Fang SG (2006b) Mitochondrial DNA variation and matrilineal structure in blue sheep populations of Helan Mountain, China. Can J Zool 84:1431–1439CrossRefGoogle Scholar
  45. Wu CP (2003) Seasonal patterns in group, population composition, daily activity cycle and habitat selection of blue sheep (Pseudois nayaur) in Helanshan. A thesis for master degree. National Sun Yat-Sen University, TaiwanGoogle Scholar
  46. Wu Y, Yuan CG, Hu JC, Peng JT, Tao PL (1990) A biological study of dwarf blue sheep. Acta Theriol Sin 10:185–188Google Scholar
  47. Yang JK (2001) Phylogenetic relationships and biogeography of Pseudois nayaur szechuanensis (Artiodactyla: Bovidae) based on mtDNA sequences and cranial morphometrics. A thesis for master degree. National Sun Yat-Sen University, TaiwanGoogle Scholar
  48. Yang S, Yin Z, Ma X, Lei F (2006) Phylogeography of ground tit (Pseudopodoces humilis) based on mtDNA: evidence of past fragmentation on the Tibetan Plateau. Mol Phylogenet Evol 41:257–265PubMedCrossRefGoogle Scholar
  49. Zeng B, Xu L, Yue B, Li Z, Zou F (2008) Molecular phylogeography and genetic differentiation of blue sheep Pseudois nayaur szechuanensis and Pseudois schaeferi in China. Mol Phylogenet Evol 48:387–395PubMedCrossRefGoogle Scholar
  50. Zhang D, Feng Q, Jian M (2000) Eco-environmental effects of the Qinghai-Tibet plateau uplift during the Quaternary in China. Environ Geol 39:1352–1358Google Scholar
  51. Zheng H, Powell CM, An ZS, Dong G (2000) Pliocene uplift of the northern Tibet Plateau. Geology 8:715–718CrossRefGoogle Scholar
  52. Zheng B, Xu Q, Shen Y (2002) The relationship between climate change and Quaternary glacial cycles on the Qinghai-Tibetan plateau: review and speculation. Quatern Int 97–98:93–101CrossRefGoogle Scholar
  53. Zhou CQ, Zhou KY, Hu JC (2003) The validity of the dwarf bharal (Pseudois schaeferi) species status inferred from mitochondrial Cyt b gene. Acta Zool Sin 49:578–584Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Shuai Tan
    • 1
  • Dandan Zou
    • 1
  • Lei Tang
    • 1
  • Gaochao Wang
    • 1
  • Quekun Peng
    • 1
  • Bo Zeng
    • 1
  • Chen Zhang
    • 1
  • Fangdong Zou
    • 1
    Email author
  1. 1.Sichuan Key Laboratory of Conservation Biology on Endangered Wildlife, Key Laboratory of Bio-resources and Eco-environment (Ministry of Education), College of Life SciencesSichuan UniversityChengduPeople’s Republic of China

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