In order to analyze the spatial distribution characteristics of the cultural landscape of the Yangtze River (Nanjing section) and show the role of the Yangtze River culture in promoting regional economic development and cultural radiation, Based on the Knowledge graph technology and 3S technology, this paper conducts geographical positioning and spatial analysis of the Yangtze River cultural landscape in Nanjing, and analyzes the surrounding environment by combining remote sensing images and topographic data. Researches show that: The Yangtze River cultural landscape in Nanjing includes 74 landscape points of seven landscape types, and the distribution of landscape points shows a clustering trend, but the spatial distribution and aggregation degree of different types of cultural landscapes are different. Due to the influence of climate environment and water resources distribution, the surrounding cultural landscape has a low altitude and high vegetation coverage, showing obvious distribution characteristics along the river. This study is helpful to improve the coordination degree between cultural landscape protection and planning management, and can provide useful reference for the protection of cultural landscape of the Yangtze River.