DNA dataset matrix

DNA dataset matrix in nexus format with four genes (COI, H3, 16S, 18S), 2111 characters and 38 taxa

Phylogeny from Bayesian analysis

Majority-rule consensus tree from bayesian analysis with MrBayes 3.2 in nexus format

Supplementary Tables

Supplementary Table 1. Primers used for PCR amplification and sequencing.Supplementary Table 2. General properties of the DNA matrix. Invar = number of invariant sites. Var = number of variable sites. PI = number of parsimony informative sites. A, T, C, G = proportion of respective base

<i>Agabus bipustulatus</i> classification of secondary elytral reticulation after the total sample was screened for common patterns.

<p>Seven categories (A-G) ranging from tightly packed longitudinally elongated meshes to more or less isomorphic meshes were recognised in <i>Agabus bipustulatus</i>. Type A is found in the <i>bipustulatus</i> and <i>solieri</i> forms, type B in <i>kiesenwetterii</i>, <i>falcozi</i> and <i>dolomitanus</i>, and type C, F and G in <i>pyrenaeus</i>. The categories D, E and F describe intermixed patterns.</p

The evolutionary history of the <i>Colymbetes paykulli</i> complex was inferred by using the Maximum Likelihood method based on the Hasegawa-Kishino-Yano model with a discrete Gamma distribution (5 categories (+<i>G</i>, parameter = 0.3301)) and a invariable rate variation model ([+<i>I</i>], 58.5832% sites).

<p>The tree with the highest log likelihood (-1493.9031) is shown. Bootstrap values above 60% are reported below branches. Above branches are the Maximum likelihood partition support values from the Poisson tree processes (PTP) model given for each species identified by the model. Sequence reference to individual sample from population follow <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0143577#pone.0143577.t002" target="_blank">table 2</a>. The Nearctic <i>C</i>. <i>paykulli</i> is within this study accepted as a valid species and should be known as <i>C</i>. <i>longulus</i> LeConte.</p

Sampled species and collection sites along with their respective locality code.

<p>Collectors of the populations/specimens are: Anders Nilsson (AN), Bertil Andrén (BA), Carmen E. Sainz Cantero (CSC), Ignacio Ribera (IR), Johannes Bergsten (JB), Marcus K. Drotz (MD), Stefan Ericsson (SE), and Shidi O. Hosseinie (SH). Map codes are given for <i>A. bipustulatus</i> collection sites, which are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009034#pone-0009034-g002" target="_blank">figure 2</a>. Collection sites in bold were used both in the morphological and population genetic studies.</p

Distribution map of reported observations of the Holarctic (Δ) <i>Colymbetes dahuricus</i>, the Palearctic (●) <i>C</i>. <i>paykulli</i> and the Nearctic (■) <i>C</i>. <i>paykulli</i>.

<p>The latter is within this study accepted as a valid species. This means that the name of the Palearctic species is <i>C</i>. <i>paykulli</i> Erichson and the Nearctic species should be known as <i>C</i>. <i>longulus</i> LeConte.</p

Summary of genetic variability in <i>Agabus bipustulatus</i>.

<p>Including mean number of successfully scored individuals per population over all loci, mean number of alleles per locus including ±1 standard error (SE), mean observed heterozygosity estimated as direct count including ±1 SE, and inbreeding coefficient per population measured by <i>F</i>_is. Locality codes follow <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009034#pone-0009034-t001" target="_blank">table 1</a>.</p

Maximum likelihood representation of sequence data from <i>Agabus bipustulatus</i> complex.

<p>Likelihood representation including all three genes (cytochrome b, cytochrome c oxidase subunit I and cytochrome oxidase II). The translational model (TIM) was used along with a proportion of invariable sites equal to 0.8199 and a gamma distribution shape parameter of 0.7877. The tree is rooted with the <i>Agabus bipustulatus</i> specimen from BipRus1 in order to visualise the deep split within the complex (group I and II). Jackknife values above 50% are reported above branches.</p

<i>Agabus bipustulatus</i> collecting sites.

<p>Map codes and corresponding locality codes are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009034#pone-0009034-t001" target="_blank">table 1</a>.</p