Abstract
The taxonomic status of the species Lepus melainus, the Manchurian black hare, is intensely debated. It is considered either as a valid species or a black color morph of L. mandshuricus, the Manchurian hare. Herein, we evaluate the validity of L. melainus using 24 morphological traits and two nuclear DNA loci (TG=466bp; MGF=592bp) from newly collected specimens. Except for winter pelage, we fail to discover significant morphological differences between L. melainus and L. mandshuricus. Analysis of the nuclear DNA sequences reveals lack of reciprocal monophyly between L. mandshuricus and L. melainus, as they form one single clade with high bootstrap support; in addition, morphometric and morphological analyses found no specific differentiation between forms corresponding to L. mandshuricus or L. melainus. Together with the fact that the range of L. melainus is completely within that of L. mandshuricus, our study supports the recognition of L. melainus as a melanistic morph and junior synonym of L. mandshuricus.References
Akaike, H. (1974) A new look at the statistical model identification. IEEE Transactions on Automatic Control, 19, 716–723.
Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D.J. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25, 3389–402.
Alves, P.C., Ferrand, N., Suchentrunk, F. & Harris, D.J. (2003) Ancient introgression of Lepus timidus mtDNA into L. granatensis and L. europaeus in the Iberian Peninsula. Molecular Phylogenetics and Evolution, 27, 70–80.
Alves, P.C., Harris, D.J., Melo-Ferreira, J., Branco, M., Suchentrunk, F., Boursot, P. & Ferrand, N. (2006) Hares on thin ice: introgression of mitochondrial DNA in hares and its implications for recent phylogenetic analyses. Molecular Phylogenetics and Evolution, 40, 640–641.
Alves, P.C., Melo-Ferreira, J., Freitas, H. & Boursot, P. (2008) The ubiquitous mountain hare mitochondria: multiple introgressive hybridization in hares, genus Lepus. Philosophical Transactions of the Royal Society B: Biological Sciences, 363, 2831–2839.
Ballard, J.W.O. & Whitlock, M.C. (2004) The incomplete natural history of mitochondria. Molecular Ecology, 13, 729–744.
Ellerman, J.R. & Morrison-Scott, T.C.S. (1951) Checklist of Palaearctic and Indian Mammals, 1758–1946. Bulletin of the British Museum (Natural History), London, pp. 419–445.
Estonba, A., Solis, A., Iriondo, M., Sanz-Martin, M.J., Perez-Suarez, G., Markov, G. & Palacios, F. (2006) The genetic distinctiveness of the three Iberian hare species: Lepus europaeus, L. granatensis, and L. castroviejoi. Mammalian Biology, 71, 52–59.
Felsenstein, J. (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783–791.
Flux, J.E.C. & Angermann, R. (1990) The hares and jackrabbits. In: Chapman, J.A. & Flux, J.E.C. (Eds.), Rabbits, Hares and Pikas: Status Conservation Action Plan. International Union for Conservation of Nature and Natural Resources. Gland, Switzerland, pp. 61–94.
Freitas, H. (2006) Natural hybridization between the Iberian hare (Lepus granatensis) and the brown hare (L. europaeus) in northern Iberian Peninsula. MSc thesis, Faculdade de Ciências do Porto, Porto, Portugal.
Hoffmann, R.S. (1993) Family Leporidae. In: Wilson, D.E. & Reeder, D.M. (Eds), Mammal Species of the World: A Taxonomic and Geographic Reference, 2nd ed. Smithsonian Institution Press. Washington, DC, pp. 818–819.
Hoffmann, R.S. & Smith, A.T. (2005) Family Leporidae. In: Wilson, D.E. & Reeder, D.M. (Eds), Mammal Species of the World: A Taxonomic and Geographic Reference, 3rd ed. Johns Hopkins University Press. Baltimore, pp. 200–201.
Inc, S. (2004) SPSS 13.0 Base User's Guide. SPSS Inc., Chicago, Illinois.
Li, Z.Y. & Luo, Z.X. (1979) A new species: Lepus melainus. The Transactions of Northeast Forest University, 2, pp. 71–81. [In Chinese.]
Loukashkin, A.S. (1943) On the hares of northern Manchuria. Journal of Mammalogy, 24, 75–81.
Luo, Z.X. (1988) The Chinese Hare. China Forestry Publishing House, Beijing. [In Chinese.]
Matthee, C.A., Burzlaff, J.D., Taylor, J.F. & Davis, S.K. (2001) Mining the mammalian genome for artiodactyl systematics. Systematic Biology, 50, 367–390.
Matthee, C.A., Vuuren, B.J.V., Bell, D. & Robinson, T.J. (2004) A molecular supermatrix of the rabbits and hares (Leporidae) allows for the identification of five intercontinental exchanges during the Miocene. Systematic Biology, 53, 433–447.
Melo-Ferreira, J., Alves, P.C., Freitas, H., Ferrand, N. & Boursot, P. (2009) The genomic legacy from the extinct Lepus timidus to the three hare species of Iberia: contrast between mtDNA, sex chromosomes and autosomes. Molecular Ecology, 18, 2643–2658.
Melo-Ferreira, J., Boursot, P., Randi, E., Kryukov, A., Suchentrunk, F., Ferrand, N. & Alves, P.C. (2007) The rise and fall of the mountain hare (Lepus timidus) during Pleistocene glaciations: expansion and retreat with hybridization in the Iberian Peninsula. Molecular Ecology, 16, 605–618.
Melo-Ferreira, J., Boursot, P., Suchentrunk, F., Ferrand, N. & Alves, P.C. (2005) Invasion from the cold past: extensive introgression of mountain hare (Lepus timidus) mitochondrial DNA into three other hare species in northern Iberia. Molecular Ecology, 14, 2459–2464.
Noack, Th. (1891) Wild und Wald in Ost-Sibirien. Der Weidmann, Bd. 22, no. 48, s. 420.
Ognev, S.I. (1922) Materials on systematics of the Russians hares. Annuaire du [Musée du]'Academie Imperial des Sciences, 22, 489. [In Russian]
Pan, Q.H., Wang, Y.X. & Yan, K. (2007) A Field Guide to the Mammals of China. China Forestry Publishing House, pp. 2–8. [In Chinese.]
Posada, D. & Buckley, T.R. (2004) Model selection and model averaging in phylogenetics: advantages of Akaike Information Criterion and bayesian approaches over likelihood ratio tests. Systematic Biology, 53, 793–808.
Posada, D. & Crandall, K.A. (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics, 14, 817–818.
Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, second ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Sawyer, S. (1989) Statistical tests for detecting gene conversion. Molecular Biology and Evolution, 6, 526–538.
Sawyer, S. (2000) GENECONV: Statistical tests for detecting gene conversion (version 1.81). Computer program distributed by the author. Washington University, St. Louis, MO, USA.
Sowerby, A. de C. (1923) The Naturalist in Manchuria. Tientsin Press Ltd., China, p. 1–191.
Swofford, D.L. (2002) PAUP*: Phylogenetic analysis using parsimony (* and Other Methods). Version 4. Sinauer Associates, Sunderland, Massachusetts.
Thompson, J.D., Gibson, T.J., Plewniak, F., Mougin, F.J. & Higgins, D.G. (1997) The Clustal X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876–4882.
Thulin, C.G., Jaarola, M. & Tegelstrǒm, H. (1997) The occurrence of mountain hare mitochondrial DNA in wild brown hares. Molecular Ecology, 6, 463–467.
Wu, C.H., Li, H.P., Wang, Y.X. & Zhang, Y.P. (2000) Low genetic variation of the Yunnan hare (Lepus comus Allen 1927) as revealed by mitochondrial Cytb gene sequences. Biochemical Genetics, 38, 149–155.
Wu, C.H, Wu, J.P, Bunch, T.D., Li, Q.W, Wang, Y.X. & Zhang, Y.P. (2005) Molecular phylogenetics and biogeography of Lepus in eastern Asia based on mitochondrial DNA sequences. Molecular Phylogenetics and Evolution, 37, 45–61.