One of the major debates in European prehistory concerns the nature of the Neolithic Transition. The Neolithic way of life, where a sedentary farming spreading from the Near East to the Central Europe replaced forager lifestyles, changed human biology, dietary choices and subsistence strategies. The Balkans represents one of the key areas for studying the process of Neolithisation, as it is located at the crossroad between the Near East and Central Europe. Results of previous dietary studies in the Central Balkan area, based on stable isotope data (C, N, S) indicate that many Neolithic humans remained reliant on aquatic resources. The speed of the Neolithisation process and the extent that the rich resources of some of the river environments within Central Balkans delayed uptake of domestic cultigens is unclear. The identification of microbotanical remains in human dental calculus is a powerful tool to access direct evidence of ancient diets. Dietary reconstructions based on plant microfossils, such as starch grains and phytoliths entrapped in calculus, have been useful in increasing our knowledge of plant consumption in ancient populations. The rich skeletal collection record from sites in the Central Balkans with its long Mesolithic-Neolithic sequence (9500-5200 cal. BC), provides the opportunity to reconstruct plant use and local subsistence practices by using dental calculus analysis. The resulting data sheds light on the nature of the Mesolithic-Neolithic transformation and provide better understanding of the dynamics of the Neolithisation process across the continent.