To elucidate the evolutionary history of chamois, we had analysed DNA sequences of four mitochondrial regions and 20 loci microsatellites including all subspecies along its entire distribution range

1. MICROSATELLITE AND mt-DNA PHYLOGENIES OF THE CHAMOIS (GENUS RUPICAPRA ) AND TAXONOMIC IMPLICATIONS OVIEDO UNIVERSITY DEPARTMENT OF FUNCTIONAL BIOLOGY Mª Trinidad Pérez Méndez

2. PRESENT DISTRIBUTION OF RUPICAPRA GENUS R. pyrenaica R. rupicapra

3. EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME Lydekker 1913 Rupicapra rupicapra Taxonomy of the genus has been object of continuos revisions. parva pyrenaica ornata cartusiana rupicapra tatrica carpatica balcanica asiatica caucasica

4. EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME Taxonomy of the genus has been object of continuos revisions. Rupicapra rupicapra Rupicapra ornata Rupicapra pyrenaica Lydekker 1913 1914 Camerano parva pyrenaica ornata cartusiana rupicapra tatrica carpatica balcanica asiatica caucasica

5. EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME Rupicapra rupicapra Taxonomy of the genus has been object of continuos revisions. Lydekker 1913 1914 Camerano Couturier 1938 Dolan 1963 parva pyrenaica ornata cartusiana rupicapra tatrica carpatica balcanica asiatica caucasica

6. EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME Taxonomy of the genus has been object of continuos revisions. Rupicapra rupicapra Rupicapra pyrenaica Lydekker 1913 Couturier 1938 Dolan 1963 Nascetti 1985 1987 Lovari 1914 Camerano parva pyrenaica ornata cartusiana rupicapra tatrica carpatica balcanica asiatica caucasica

7. GEOLOGIC TIME LINE AND GLACIAL PERIODS IN EUROPE

8. CHAMOIS PALEONTOLOGICAL DATA Caune de l´Arago - Mindel Riss’ fossils Riss-Würm’ fossils Distribution limit during Würm Chamois fossils (Masini y Lovari, 1988)

9. PHYLOGEOGRAPHY & COLONIZATION ROUTES IN EUROPE Phylogeographic analyses can evaluate the relative role of different historical forces in shaping the present genetic structure of populations. I.e.- Glaciation cycles of the Quaternary, have periodically restricted some species into disjoint refugia. Once a reversal in climate change allows for rapid migration out of refugial areas, these species spread rapidly into newly available habitat. A number of empirical studies find genetic signatures that support this scenario of refugia and postglacial expansion. Taberlet et al., 1998

11. 20 loci microsatellite and 5 mitochondrial regions are analyzed 486 bp 400 bp 484 bp 471 bp 525-584 bp Control Region mtDNA regions analyzed

13. SCORED INDIVIDUALS Total 215 samples Microsatellites 179 samples mtDNA 152 samples Both markers 116 samples Subspecies 29 11 4 Only microsatellites Both markers Only mtDNA Parva 29 11 4 Pyrenaica 15 26 0 Cartusiana 0 8 0 ornata 0 12 0 rupicapra 2 29 20 tatrica 0 10 0 carpatica 7 10 6 caucasica 4 6 0 asiatica 0 1 0 balcanica 6 3 6

14. Haplotypic diversity % Nucleotidic diversity % Microsatellite dataset 184 alleles 9.20 alleles/locus ***P<0.001 Mitochondrial combined dataset 1646 nt 79 haplotypes 239 variable sites 196 parsimony-informative He vs Ho % POPULATION DIVERSITY *** *** - - 100 80 60 40 20 0

15. INDIVIDUAL MICROSATELLITE N-J TREE pyrenaica parva caucasica rupicapra balcanica carpatica tatrica cartusiana ornata asiatica

16. MICROSATELLITE GENETIC DISTANCES (  ) 2 VS GEOGRAPHIC DISTANCES Distancia geográfica en Km.   Geographical distance in Km. (  ) 2 Between subpopulations Between R. pyrenaica populations ( AN-Others) Between R. rupicapra populations Between species

17. INDIVIDUAL mtDNA N-J TREE East Central West

18. MITOCHONDRIAL DNA PHYLOGENY OF CHAMOIS Clade West Clade Central Clade East 1/1/0.71/1 1/1/0.97/1 1/1/0.72/1 0.51/0.50/-/0.55

19. NETWORK OF mtDNA HAPLOTYPES Clade East Clade Central Clade West

20. DISCORDANT PHYLOGENIES: MICROSATELLITE & mtDNA Clade East Clade Central Clade West R. pyrenaica R. rupicapra R. pyr. ornata

21. BEAST & BEAUti (MY: million years ) Bovidae-Caprinae: 25,8 MY± 0,6 Caprinae: 14,1 MY ± 1,1 Capra-Ovis : 11,5 MY ± 0,9 Hernández y Vrba, 2005. Clade West Clade Central Clade East DIVERGENCY TIMES BETWEEN CLADES PLEISTOCENE

22. time Holocene: present PHYLOGEOGRAPHY AND EVOLUTIONARY HISTORY Pleistocene glacial-interglacial periods

23. RELATED BIBLIOGRAPHY Perez Trinidad; Hammer Sabine E.; Albornoz Jesus; et al. (2011) Y-chromosome phylogeny in the evolutionary net of chamois (genus Rupicapra ) .BMC EVOLUTIONARY BIOLOGY 11: 272 Rodriguez Fernando; Perez Trinidad; Hammer Sabine E.; et al. (2010) Integrating phylogeographic patterns of microsatellite and mtDNA divergence to infer the evolutionary history of chamois (genus Rupicapra ). BMC EVOLUTIONARY BIOLOGY 10: 222 Rodriguez Fernando; Hammer Sabine; Perez Trinidad; et al. (2009) Cytochrome b Phylogeography of Chamois ( Rupicapra spp.). Population Contractions, Expansions and Hybridizations Governed the Diversification of the Genus. JOURNAL OF HEREDITY 100:47-55 Rodriguez Fernando; Albornoz Jesus; Dominguez Ana (2007) Cytochrome b pseudogene originated from a highly divergent mitochondrial lineage in genus Rupicapra . JOURNAL OF HEREDITY 98: 243-249 Perez T; Albornoz J; Dominguez A (2002) Phylogeography of chamois ( Rupicapra spp.) inferred from microsatellites MOLECULAR PHYLOGENETICS AND EVOLUTION 25: 524-534 Perez T; Albornoz J; Dominguez A (2000) A panel of bovine and caprine microsatellites suitable as markers in chamois ANIMAL GENETICS 31: 344-345 Perez T; Albornoz J; GarciaVazquez E; et al. (1996) Application of DNA fingerprinting to population study of chamois ( Rupicapra rupicapra ). BIOCHEMICAL GENETICS 34: 313-320

24. RESEARCH TEAM Fernando Rodriguez Jesús Albornoz Ana Domínguez Sabine Hammer ( University of Veterinary Medicine Vienna, Austria) To all the people and institutions that had contributed to the recollection of the samples over more than ten years… Jose Francisco Quiros, Javier Pérez-Barbería, Carlos Nores, L. Rossi, Juan Carlos del Campo, Juan Bejar, Paloma Barracina, H. Papaioannou, M. Brown, W. Gasienica-Byrcyn, T. Skalski, J. Meana, S. Erceg, P. Veinberg, J. Badridge, Alvaro Mazón, Michal Adamec, Richard Kraft , Juan Herrero, Athanassios I. Sfougaris, Friederike Spitzenberger, Rita Lorenzini, Jacques Michallet, Natalia Martinkova, Franz Suchentrunk. Institutions: the Regional Governments of Principado de Asturias (Consejeria de Agricultura) and Aragón (Diputación General de Aragón), the hunting wardens from Asturias and Aragón, Camino Real Hunting.