Abstract: The spatial heterogeneity of landscape and ecosystem services can be made clear by using urban-rural gradient recognition based on landscape clustering. Here, we consider the main urban area of Nanchang with significant differences in urban and rural geographical environments, and apply an urban-rural gradient classification method based on landscape clustering. We classify 831 village-scale units, correlate the classification results with the ecosystem service values calculated by the equivalent factor method, and analyze the urban-rural gradient characteristics of four ecosystem service values. The results show that: (1) Five gradient types are dominant (urban, suburban village, exurban village, agricultural village and ecological village), and the landscape structure of each type of village is significantly different. (2) Different types of villages in the main urban area of Nanchang have different spatial locations and agglomeration characteristics, which adapt to the characteristics of local urban and rural geographical environments. Urbanization, agricultural activities and natural environment jointly drive the urban-rural gradient differentiation in the main urban area. (3) The habitat quality and carbon sequestration services in the main urban area can be divided into two growth stages along the urban-rural gradient. The food production and recreation areas are characterized by inverted ‘U’ and ‘V’ gradients, respectively. The relationship between food supply and recreation areas and carbon sequestration services in the urban-rural gradient is dominated by trade-offs. The relationship between habitat quality and recreation areas is dominated by synergy, and the habitat quality and food supply services are synergistic, and trade-off dominated in agricultural villages and ecological villages, respectively. The analysis of urban-rural gradients and their ecosystem service responses yields a deeper understanding of the spatial heterogeneity of ecosystem services, which is conducive to the improvement, appreciation, and synergetic management of urban-rural ecosystem services.