MICROSATELLITE AND mt-DNA PHYLOGENIES OF THE CHAMOIS (GENUS

RUPICAPRA) AND TAXONOMIC IMPLICATIONS

OVIEDO UNIVERSITYDEPARTMENT OF FUNCTIONAL BIOLOGY

Mª Trinidad Pérez Méndez

PRESENT DISTRIBUTION OF RUPICAPRA GENUS

R. pyrenaica R. rupicapra

EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME

parva

pyrenaica

ornata

cartusiana

rupicapra

tatrica

carpatica

balcanica

asiatica

caucasica

Lydekker1913

Rupicapra rupicapra

Taxonomy of the genus has been object of continuos revisions.

EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME

parva

pyrenaica

ornata

cartusiana

rupicapra

tatrica

carpatica

balcanica

asiatica

caucasica

Taxonomy of the genus has been object of continuos revisions.

Rupicapra rupicapra

Rupicapra ornata

Rupicapra pyrenaica

Lydekker1913

1914Camerano

EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME

parva

pyrenaica

ornata

cartusiana

rupicapra

tatrica

carpatica

balcanica

asiatica

caucasica

Rupicapra rupicapra

Taxonomy of the genus has been object of continuos revisions.

Lydekker1913

1914Camerano

Couturier1938

Dolan1963

EVOLUTION OF Rupicapra GENUS CLASIFICATION OVER TIME

parva

pyrenaica

ornata

cartusiana

rupicapra

tatrica

carpatica

balcanica

asiatica

caucasica

Taxonomy of the genus has been object of continuos revisions.

Rupicapra rupicapra

Rupicapra pyrenaica

Lydekker1913

Couturier1938

Dolan1963

Nascetti1985

1987Lovari

1914Camerano

GEOLOGIC TIME LINE AND GLACIAL PERIODS IN EUROPE

CHAMOIS PALEONTOLOGICAL DATA

Caune de l´Arago - Mindel

Riss’ fossils

Riss-Würm’ fossils

Distribution limit during Würm

Chamois fossils (Masini y Lovari, 1988)

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

MAIN GOALS

Study of the evolutionary history of the genus

Revision of chamois taxonomy

Evolutionary Significant Units (ESU).

Equivalent to species under the Phylogenetic Species Concept (Vogler y

DeSalle, 1994).

Conspecific populations which are reciprocally monophyletic for mtDNA

alleles and differ significantly for the frequency of alleles at nuclear loci

(Moritz, 1994).

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

• Microsatellite markers are inherited in a Mendelian way

meanwhile mtDNA is transmitted predominantly through maternal

lineages in most species.

• Microsatellite markers have a higher evolution rate than mtDNA.

• Microsatellite markers narrate the recent phylogenetic history of

tens or thousands of years while mitochondrial ones shed light on

the deeper phylogenetic history.

MICROSATELLITE DNA vs. mtDNA

SCORED INDIVIDUALS

Total 215 samplesMicrosatellites 179 samplesmtDNA 152 samples Both markers 116 samples

Subspecies

29 11 4

Only microsatellitesBoth 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

Haplotypicdiversity %

Nucleotidicdiversity %

Microsatellite dataset184 alleles9.20 alleles/locus

***P<0.001

Mitochondrial combined dataset1646 nt79 haplotypes239 variable sites196 parsimony-informative

He vs Ho%

POPULATION DIVERSITY

***

***

- -

100

80

60

40

20

0

INDIVIDUAL MICROSATELLITE N-J TREE

pyrenaica

parv

a

caucasica

rupicapra

balcan

ica

carpatica

tatri

ca

cartusiana

ornata

asiatica

MICROSATELLITE GENETIC DISTANCES

()2 VS GEOGRAPHIC DISTANCES

Distancia geográfica en Km.

Between subpopulations

Between R. pyrenaica populations ( AN-Others)

Between R. rupicapra populations

Between species

Geographical distance in Km.

()

2

INDIVIDUAL mtDNA N-J TREE

East

Central

West

MITOCHONDRIAL DNA

PHYLOGENY OF

CHAMOIS

1/1/0.71/1

1/1/0.97/1

1/1/0.72/1

0.51/0.50/-/0.55

CladeWest

Clade Central

Clade East

NETW

OR

K O

F m

tDN

A H

AP

LO

TY

PES

Clade East

Clade Central

Clade West

Clade East

Clade Central

Clade West

R. p

yren

aica

R. rupicapra

R. pyr. ornata

DISCORDANT PHYLOGENIES: MICROSATELLITE & mtDNA

BEAST & BEAUti (MY: million years)Bovidae-Caprinae: 25,8 MY± 0,6Caprinae: 14,1 MY ± 1,1Capra-Ovis: 11,5 MY ± 0,9Hernández y Vrba, 2005.

CladeWest

CladeCentr

al

Clade East

DIVERGENCY TIMES BETWEEN CLADESPLEISTOCENE

time

Holo

cene:

pre

sent

PHYLOGEOGRAPHY AND EVOLUTIONARY

HISTORY

Ple

isto

cen

e g

laci

al-

inte

rgla

cial peri

ods

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

RELATED BIBLIOGRAPHY

RESEARCH TEAM

Fernando RodriguezJesús AlbornozAna 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.