Mountain ranges may delimit the distribution of native species as well as constitute potential barriers to the spread of invasive species. The invasive pinewood nematode, Bursaphelenchus xylophilus, is a severe forest pest inducing pine wilt disease. It is vectored in Europe by a native long-horned beetle, Monochamus galloprovincialis. This study explored the potential of the Pyrenean chain to slow or prevent the natural spread of nematode-infested beetles from the Iberian Peninsula, where the nematode is established and is expanding its range, towards France and the rest of Europe. An analysis of the genetic structure and migration patterns of the beetle populations throughout the Pyrenean mountain range was combined with a spread model simulating the potential movements of nematode-infested beetles across it. The central part of the Pyrenees, which corresponds to the highest elevation zone, was shown to prevent gene flow between the French and Spanish populations of M. galloprovincialis on each side of the mountains. Conversely, strong admixture was detected between populations located on both sides of low elevation hills, and especially at the east and west extremities of the mountain range. Simulations of the spread of nematode-infested beetles under various thresholds of beetle survival and pine wilt disease expression gave results consistent with the variation in genetic make-up, suggesting that western and eastern hillsides may represent corridors favoring natural spread of the nematode from the Iberian Peninsula to France. Simulations also showed that temperature rise due to climate change may significantly reduce the extent of the barrier formed by highest elevations. Our results support the hypothesis that the Pyrenean chain represents a partial barrier to the natural spread of nematode-infested beetles. These results, which have to be considered together with potential human-assisted long-distance spread of the nematode, highlight priority zones for future pest monitoring and management programs. More generally, such an integrated approach could be used to assess the role of mountain chains in the potential spread of other invasive pests.