Landscape genetics have been particularly relevant when assessing the influence of landscape characteristics on the genetic variability and the identification of barriers to gene flow. Linking current practices of area-wide pest management information on pest population genetics and geographical barriers would increase the efficiency of these programs. The woolly apple aphid, Eriosoma lanigerum (Hausmann), an important pest of apple orchards worldwide, was collected on apple trees (Malus domestica Borkh) from different locations in a 400 km north-south transect trough central Chile. In order to determine if there was population structure, diversity and flow were assessed. A total of 215 individuals from these locations were analysed using Inter Simple Sequence Repeat (ISSR) markers. Four ISSR primers generated a total of 114 polymorphic loci. The percentage of molecular variation among locations was 18%. As the algorithm used by structure may be poorly suited for inferring the number of genetic clusters in a data set that has an IBD relationship, the number of genetic clusters in the samples was also analyzed using a Bayesian clustering method implemented in software Baps version 4.14. We inferred the presence of four genetic clusters in the study region. Clustering of individuals followed a pattern explained by some geographical barriers. Using partial Mantel tests, we detected barriers to gene flow other than distance, created by a combination of main rivers and mountains. Although landscape genetics are rarely used in pest management, our results suggest that these tools may be suitable for the design of area-wide pest management programs.