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Supplemental Information

Detailed information for all analysed individuals

Table S1. All analysed individuals with BOLD process ID, sample ID, Barcode Index numbers (BIN) and their COI haplotype membership, primers pairs used for amplification.

DOI: 10.7287/peerj.preprints.2741v1/supp-1

Divergence time for key nodes in the time-calibrated reconstruction of phylogeny

Table S2. Divergence time (Ma) for key nodes and rates of COI evolution in substitutions per site, per My estimated using Bayesian inference for different calibration schemes. Node affiliation given in Fig. S1. Calibration schemes based on secondary calibration points, standard rate and geological points (see material and methods). Lower and upper 95% highest posterior densities (95%HPD) are provided.

DOI: 10.7287/peerj.preprints.2741v1/supp-2

Results of the Bayesian node reconstruction

Table S3. Node ages (Ma) with lower and upper 95% highest posterior densities, geographical coordinates of the ancestral locations expressed as mean and standard deviation (SD), probabilities associated with occurrence in riverine (P(R)) or in lacustrine (P(L)) habitat (see Figure S1 for node identification).

DOI: 10.7287/peerj.preprints.2741v1/supp-3

K2p distances between MOTUs

Table S4. Mean Kimura two parameters (K2p) distances (below the diagonal) and standard error (SE) (above the diagonal) between MOTUs for 74 COI haplotypes. N and k = number of individual sampled and haplotypes per MOTU, respectively.

DOI: 10.7287/peerj.preprints.2741v1/supp-4

Fit of the diversification models to the Bayesian reconstruction of phylogeny

Table S5. Fit of the diversification models to Bayesian reconstruction of phylogeny, based on the Akaike information criterion (AIC).

DOI: 10.7287/peerj.preprints.2741v1/supp-5

Molecular genetic diversity and historical demography within MOTUs

Table S6. Molecular genetic diversity and historical demography based on mtDNA COI gene region (530 bp) within the 13 MOTUs (A-M). Analysis was done either at the scale of the entire MOTU (in bold) or a site or set of sites (in italic) within a MOTU.

DOI: 10.7287/peerj.preprints.2741v1/supp-6

Genetic differentiation between sites

Table S7. Genetic differentiation based on mtDNA COI gene region (530 bp) between sites (S1 vs S2) within MOTUs (A, C, E, G and K) present in more than one site within the morphospecies Gammarus roeselii in the Balkans.

DOI: 10.7287/peerj.preprints.2741v1/supp-7

Figure S1

Bayesian chronogram with node definition, calibration points and results of species delimitation methods.

DOI: 10.7287/peerj.preprints.2741v1/supp-8

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Michał Grabowski conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Tomasz Mamos conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Karolina Bącela-Spychalska conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, reviewed drafts of the paper.

Tomasz Rewicz conceived and designed the experiments, performed the experiments, prepared figures and/or tables, reviewed drafts of the paper.

Remi A Wattier conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

DNA Deposition

The following information was supplied regarding the deposition of DNA sequences:

GenBank

Accession numbers for mt COI: KP789673, KP789674, KP789675, KP789676, KP789677, KP789678, KP789679, KP789680, KP789681, KP789682, KP789683, KP789684, KP789685, KP789686, KP789687, KP789688, KP789689, KP789690, KP789691, KP789692, KP789693, KP789694, KP789695, KP789696, KP789697, KP789698, KP789699, KP789700, KP789701, KP789702, KP789703, KP789704, KP789705, KP789706, KP789707, KP789708, KP789709, KP789710, KP789711, KP789712, KP789713, KP789714, KP789715, KP789716, KP789717, KP789718, KP789719, KP789720, KP789721, KP789722, KP789723, KP789724, KP789725, KP789726, KP789727, KP789728, KP789729, KP789730, KP789731, KP789732, KP789733, KP789734, KP789735, KP789736, KP789737, KP789738, KP789739, KP789740, KP789741, KP789742, KP789743, KP789744, KP789745, KP789746

Accession numbers for 28S rDNA: KP789747, KP789748, KP789749, KP789750, KP789751, KP789752, KP789753, KP789754, KP789755, KP789756, KP789757, KP789758, KP789759, KP789760, KP789761, KP789762, KP789763, KP789764, KP789765, KP789766, KP789767, KP789768, KP789769

Data Deposition

The following information was supplied regarding data availability:

The raw data has been supplied as a supplementary file.

Funding

The sampling was performed during the Amphi-Balkan Expeditions II–V organised by the senior author and financed partially from the internal funds of the University of Lodz. This study was financially supported by the Polish Ministry of Science and Education, grant number N N303 579439. The sampling was performed during the Amphi-Balkan Expeditions II-V organised by the senior author and financed partially from the internal funds of the University of Lodz. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.


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