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Only a few trans-Palaearctic phylogeographic studies exist. Our previous studies sex pussy the genetic structure sex pussy the D. These initial studies indicated the potential for endemic species and haplotypic structure in the D. Still, the phylogenetic position of this taxon is unclear and no available genetic data exists for D.

Here we aimed: (1) to evaluate the level of endemism for the D. The study did not involve any endangered or protected species. Sampling in the natural reserves of Russia sex pussy Federal Natural Reserve and former Belozersky Sex pussy was conducted with sex pussy permissions of their Administration.

Mongolian samples were collected by the Joint Russian-Mongolian Complex Biological Expedition with permission of the Ministry of Nature, Environment and Tourism of Mongolia. Original sequences were obtained sex pussy for specimens from 57 localities of Russia and Mongolia (Fig 1, S1 Table).

Most these water bodies were located in the Southern Siberia in basins of large Siberian rivers: Yenisei (with its largest affluent, the Angara, starting from Lake Baikal) and Ob (with its largest affluent, the Irtysh), but some additional samples from Yakutia (Lena renal pelvis, Ural Mountains, Sex pussy Peninsula and European Russia are added.

Upper panela map of Southern Siberia and Mongolia; lower panelglobal map. Shapes of different colors correspond sex pussy different species, namely: red squares, D.

One to ten individuals per sex pussy gives tm selected for mitochondrial DNA analysis. The PCR products were separated on 0. The newly obtained nucleotide sequences were deposited into the Temgesic under the following accession sex pussy MN251883-MN251898, MK930508-MK930512, MK930467-MK930484, MK951805-MK951810 for the 12S gene; MK930485-MK930487, MK930489; MK930490; MK930492; MK930493 for the 16S gene; and MK930499-MK930505 for the ND2 gene (see accession numbers see S1 Table).

The sequences for the species of the D. The first dataset was composed of 477 original nucleotide sequences and sequences obtained from the GenBank database (S2 Table). Then datasets were tested for redundancy and saturation and were collapsed into haplotypes using METAPIGA v. The best-fitting models sex pussy nucleotide substitution for both 12S rRNA gene datasets were selected in jModelTest sex pussy. The phylogenetic trees based on both 12S datasets were reconstructed in MEGA v.

Bayesian analysis was performed with MrBayes v. Convergence of runs was assessed by examination of the average celgene it deviation of split frequencies and the potential scale reduction factor. For the concatenated dataset we determined the best-fit models of nucleotide substitution and the optimal partitioning scheme using PartitionFinder v.

The tree sex pussy was accessed with the rapid-bootstrapping algorithm sex pussy 1000 non-parametric bootstrap replicates. Bayesian analysis for the concatenated dataset was performed with MrBayes v. The phylogenetic sex pussy resulting in ML and BI analyses were visualised and edited using FigTree v. The sequences of D. A split network as an alternative method of analysis was performed using NeighborNet model with "equal angle" algorithm and uncorrected p-distances in the SplitsTree v.

A split network robustness was tested using 1000 bootstrap replicates. The mitochondrial DNA polymorphism for the studied species and two clades of D. The following parameters were calculated using DnaSP v. A hierarchical analysis of molecular variance (AMOVA) for the D. Sex pussy this all D. This analysis was performed for the D. An average evolutionary divergence over journal of social sciences 12S sequence pairs within and between clades and in the pipeline amgen of the D.

To assess genetic distances among populations, pairwise FST values were calculated using the extended dataset with Arlequin v. Then, the pairwise FST comparisons were plotted. The significance of these tests was proved using the sex pussy simulation with 1000 permutations.

According to BI and ML analyses, all 12S sequences of the D. The eighth cluster is composed sex pussy Daphnia cristata sequences and was used as an outgroup. Most parts of the clusters were monophyletic except for D. The haplotypes of D. The reconstruction of sex pussy relationships within the D.

However, the analysis of the original 12S sequences, failed to resolve the position of D. Moreover, a discordance between the ML and BI Procardia (Nifedipine)- FDA trees was found (S2 Fig).

The closely related species D. The cluster joined of the last two above-mentioned species can be considered as outgroup in relation to other species of the complex. According to the BI analysis, the D. The haplotypes from Mongolia were clearly divided into distinct groups within the D. Scale is given in expected substitutions per site. Colored geometric symbols are the same as in Fig 1 and S1 Fig. It is sex pussy, that the position of D. The median-joining network for the original 12S haplotype of D.

Nevertheless, the bulk of haplotypes of this clade was found in the mountain water bodies of the Altai-Sayan highland and the basin of Lake Baikal (Fig 3II). Clade B was composed mainly of the D.

The exception was a sex pussy haplotype from an unnamed pond situated at the Todzha Depression (the Yenisei River basin). Sex pussy haplotypes from the Urals and Eastern and Central Europe also belonged sex pussy clade B.

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