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Genetic Analysis of Spermophilus Undulatus Subspecies Differentiation

Explore genetic variability and relationships among Spermophilus undulatus subspecies to clarify species subdivision and taxonomy. Analyses include RAPD and mitochondrial DNA studies. Results indicate a distinct position for S.u. eversmanni. Supported by RFBR and FEBRAS grants.

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Genetic Analysis of Spermophilus Undulatus Subspecies Differentiation

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  1. DIFFERENTIATION AND SYSTEMATICS IN THE LONG-TAILED GROUND SQUIRREL, SPERMOPHILUSUNDULATUS (SCIURIDAE, RODENTIA) Marina V. Tsvirka, Vladimir P. Korablev, Marina V. Pavlenko Institute of Biology and Soil Science FEB RAS, Vladivostok, 690022, Russia

  2. Map of the ground squirrels distribution in the Palaearcticregion

  3. PROBLEM • Genus Spermophilus • Subgenus Urocitellus • Species Spermophilus undulatus • Subspecies • S. u. undulatus, • S. u. intercedens, • S. u. menzbieri, • S. u. jacutensis, • S. u.stramineus, • S. u. eversmanni two geographical forms “eastern” “western” (Vorontsov et al., 1980; Frisman, Vorontsov, 1989) (Gromov, Erbaeva, 1995)

  4. Map of S. undulatus distributions western form eastern form hybrid zone ? species or semispecies

  5. Morphometric data(Popkov, 1978) • Morphotypical study(Linetskiy, Linetskaya 1989) • Karyological data(Korablev, 1988)

  6. THE AIM IS to better understand species subdivision in S. undulatus , and ultimately speciation, in Spermophilus.

  7. THE MAIN TASKSARE • To study genetic variability and differentiation of S. undulatus based on nuclear (RAPD) and mitochondrial (D-loop) DNA. • To assess genetic divergence and phylogenetic relationships among subspesies of S. undulatus. • To specify the taxonomic rank of eversmanni (independent species S. eversmanni or subspecies of S. undulatus) based on the data obtained

  8. MATERIALS Map of sampling localities western form eastern form S. relictus, S. dauricus and S. parryi, S. taurensis and S. xanthoprymnus were chosen for comparative analyses, Tamias sibiricus was chosen as outgroup taxon.

  9. METHODS • Mitochondrial DNA analysis • 5’ end of the D-loop(518 bp) • Calculation of genetic distances based on Kimura two-parametric (K2P) estimator. • Reconstructions: NJ, MP, ML and UPGMA with bootstrap estimation of branching robustness in MEGA 4.0 (Tamura et al., 2007) RAPD-PCR analysis • 166 binary characters including the molecular markers • Ten arbitrary decanucleotide primers • Calculation of genetic distances (Nei, 1972) • Reconstructions: NJ and UPGMA with bootstrap estimation of branching robustness in TREECONW 1.3 b (Van de Peer, De Wacher, 1994 )

  10. polymorphic profiles monomorphic profiles

  11. RESULTS Electrophoregrams of S. undulatus obtained using primers OPD-05 and OPE-20 “eastern” form common for all of subspecies eversmanni menzbiery Marker loci are indicated by arrows.

  12. RAPD analysis Genetic variability among S. undulatus subspecies N –total number of loci, Np– number of polymorphic loci, A –allele frequencies, Р – percentage of polymorphicloci (as the proportion of loci polymorphic withineach population tested).

  13. Parameters of genetic differentiation on RAPD analysis n-number of samples, DN - genetic distances (Nei, 1972), p(Ex) - probability (exact test for interpopulation differentiation (Raymond and Rousset, 1995).

  14. NJ and UPGMA treesbased on RAPD data “eastern” form eversmanni UPGMA “eastern” form eversmanni

  15. mt DNA data • 519 nucleotides; • 92 variable sites, 56 parsimony informative; • Mean pairwise transition/transversion ratios was 1.284. Estimates of mean divergence between main lineages of Spermophilus given as geneticdistances

  16. NJ and UPGMA trees based on KP2 distances calculated from mtDNA data

  17. Genetic distances for nuclear (RAPD) and mitochondrial (mtDNA) genomes

  18. CONCLUSIONS • Both nuclear and mitochondrial DNA analyses showed that the subspecies of “eastern” form were close to each other and far from subspecies of “western” form. • 2. Our molecular genetic data showedan important level of genetic divergence between the animals from Altai (S. u. eversmanni) and Far East of Russia (S. u. intercedens, S. u. menzbieri, S. u. jacutensis), similar to those observed between different Spermophilus species (S. relictus, S. dauricus, S. parryi, S. xanthoprymnus, S. taurensis). • 3. The results confirm a distinct taxonomic position of eversmanni. It should be considered as a species taxon within Urocitellus group.

  19. ACKNOWLEDGMENTS • The study was supported by the RFBR (grant nos. 06-04-39015) and FEBRAS (grant nos. 09-III-В-138). THANKS FOR YOUR ATTENTION

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