Due to the different geo-morphological, climatic, economic and social factors, Serbia represents a very heterogeneous area with specific historical legacies that are hard to overcome. Therefore, the regional specificities represent a starting point for planning the development of the economy as a whole, and of the agribusiness in particular. It is important to properly identify the regional peculiarities of agriculture in order to contribute to the agricultural development of Serbia as a whole. First of all, the attention should be placed to overcome the problems of underdeveloped areas that would contribute to a more stable and harmonious development of agriculture in Serbia. Balanced regional development policies should encourage better use of natural resources, especially in lagging behind areas. Spatial planning is a tool to create quality changes, especially in rural areas, linking different sectors (agriculture, food processing, tourism, environmental protection, etc.). The achievement of set objectives is highly dependent on the level of development of small and medium enterprises (SMEs) in agribusiness. Agribusiness is particularly interesting field for the development of SMEs as it is a complex area that involves the production and processing of agricultural and food products. Therefore, by using the appropriate mathematical and statistical methods, evaluation of the operational efficiency of SMEs in agribusiness in districts of Serbia was performed, in order to address the deficiencies and improvement opportunities in business in some areas.