Taking Jiangsu Province as an example, a rural regional function evaluation system was built from four perspectives: economic development, agricultural production, social security and ecological conservation. Based on the GIS spatial analysis method, the spatial differentiation characteristics of rural regional functions in Jiangsu Province are revealed, the trade-offs between functions are further explored and the multifunctional spatial distribution in rural areas of Jiangsu Province. The results of the study are as following three aspects: Jiangsu Province's rural regional functional space differentiation characteristics are obvious, and the economic development function presents the distribution characteristics of high in the south and low in the north. The hotspots of agricultural production are mainly located in northern Jiangsu, and distribution characteristics of the ecological conservation function is high in the north and low in the middle. Areas with strong social security functions are Nanjing, Suzhou and surrounding areas. There is a trade-off relationship between the four functions. The trade-off intensity of economic development function is the highest and social security function is the lowest. The multi-functional areas of rural areas in Jiangsu Province are mainly distributed in southern Jiangsu. The single-functional areas in rural areas are mainly distributed in central Jiangsu and some coastal areas in northern Jiangsu. The non-multifunctional area of rural areas is mainly located in the western part of northern Jiangsu and parts of Taizhou City.