The Neolithic settlement of Dispilió (lake Orestiás, northern Greece) is a unique site due to its continual inhabitation from the Middle Neolithic (5400 BC) to the Chalcolithic (3600 BC) and its exceptional location at the shore of Lake Orestiás (also known as Lake Kastoria). The plant landscape shaping of the area has been examined by correlating pollen and non-pollen palynomorphs (NPPs) from both on-site and off-site records. The examined pollen profiles bear concrete evidence of agricultural activities and land use in the area. The local pollen assemblages identified, each of them corresponding either to a different kind or intensity of exploitation of natural resources, display the diachronic evolution of the cultural landscape during more than 2000 years of human activity. The successive phases of intense human impact on vegetation patterns are in good accordance with the periods of major constructing activities described by the archaeological study of the Dispilió settlement. Finally, the differences on palynomorph accumulation pathways of the examined records made possible the distinction between local and “regional” plant landscapes and related human activities.