To achieve a sustainable development, impacts on biodiversity of urbanisation, new infrastructure projects and other land use changes must be considered on landscape and regional scales. This requires that important decisions are made after a systematic evaluation of environmental impacts. Landscape ecology can provide a conceptual framework for the assessment of consequences of long-term development processes like urbanisation on biodiversity components, and for evaluating and visualising the impacts of alternative planning scenarios. The aim of this paper was to develop methods for integrating biodiversity issues in planning and strategic environmental assessment in an urbanising environment, on landscape and regional levels. In order to test developed methods, a case study was conducted in the region of Stockholm, the capital of Sweden, and the study area embraced the city centre, suburbs and peri-urban areas. Focal species were tested as indicators of habitat quality, quantity and connectivity in the landscape. Predictive modelling of habitat distribution in geographic information systems involved the modelling of focal species occurrences based on empirical data, incorporated in a landscape ecological decision support system. When habitat models were retrieved, they were applied on future planning scenarios in order to predict and assess the impacts on focal species. The scenario involving a diffuse exploitation pattern had the greatest negative impacts on the habitat networks of focal species. The scenarios with concentrated exploitation also had negative impacts, although they were possible to mitigate quite easily. The predictions of the impacts on habitats networks of focal species made it possible to quantify, integrate and visualise the effects of urbanisation scenarios on aspects of biodiversity on a landscape level.