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Supplemental Information

Distribution of sampling sites in the Iberian Peninsula

Black dots refer to populations of size > 19 individuals. Green background refers to elevation (from pale to dark green: low to high elevation).

DOI: 10.7287/peerj.preprints.2968v1/supp-1

Number of individuals in sampling areas across spatial scal

Mean: black; +/- SD: grey

DOI: 10.7287/peerj.preprints.2968v1/supp-2

Evolution of DeltaK among an increasing number of K (2 -20)

DOI: 10.7287/peerj.preprints.2968v1/supp-3

Sampling details of the 132 demes

(Long. and Lat. refer to geographic coordinates of sampling sites; N. is the number of individuals of demes; A. mean allelic richness; AR. corrected allelic richness, accounting to variation in deme size; Fis. Fis estimate of deme, computed without Mon01 and Mon 27).

DOI: 10.7287/peerj.preprints.2968v1/supp-4

Details of primer sequence and genotyping

Multiplexed PCR were performed in a 10 µL reaction volume using 25 ng of genomic DNA, 0.4 U of DreamTaq DNA Polymerase (Thermo Scientific®), 0.75 µL Dream Taq Green Buffer (including 20 mM MgCl2, Thermo Scientific®), 1 µM Betaine, 0.24 µL dNTP (10 µM) and deionized H2O. PCR amplifications were run on a Veriti® 96 well fast Thermal cycler (Applied Biosystems®) using the following settings: a first denaturation step at 95 °C during 10 min; 40 cycles of denaturation (30 s at 95 °C), hybridization (30 s at 55 °C) and elongation (1 min at 72 °C), and a final elongation step at 72 °C during 10 min. One µL of PCR products were denatured within a mix of 10 µL of formamide and 0.3 µL of 600 Liz marker before being run on an ABI PRISM 3500 sequencer (Life Technologies®). Genotypes were read using the software GENEMAPPER V 4.1 (Applied Biosystems®).

DOI: 10.7287/peerj.preprints.2968v1/supp-5

Sampling locations and microsatellite genotypes

DOI: 10.7287/peerj.preprints.2968v1/supp-6

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

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

Jean-Pierre Rossi performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Juan Pajares conceived and designed the experiments, reviewed drafts of the paper.

Luis Bonifacio conceived and designed the experiments, reviewed drafts of the paper.

Pedro Naves conceived and designed the experiments, reviewed drafts of the paper.

Alain Roques wrote the paper, reviewed drafts of the paper.

Géraldine Roux analyzed the data, wrote the paper, reviewed drafts of the paper.

Funding

This work was supported by the European project REPHRAME KBBE.2010.1.4-09 (FP7 Project, Analysis of the potential of the pine wood nematode (Bursaphelenchus xylophilus) to spread, survive and cause pine wilt in European coniferous forests in support of EU plant health policy). First author was funded by the French Ministry of Research and Education. Field work was supported by COST Action FP1002 (COST-STSM-FP1002-14177). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.


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