Research on the meteorological observation data of conventional meteorological stations in Jiangnan area from 2001 to 2020 and digital elevation DEM, the “golden section” comfort index calculation method was used to refine the spatial estimation of climate comfort in Jiangnan area under undulating terrain on the Gis platform, and its spatiotemporal distribution characteristics in the region were analyzed. The results can provide reference for the development and utilization of climate resources in Jiangnan area. The results show that: (1) Climatic comfort in winter and summer shows a trend from low latitude to high latitude, while in summer it is opposite. (2) Climatic comfort in winter shows a significant trend from low altitude to high altitude, while in summer it is opposite. (3) The ocean has a good regulating effect on apparent temperature in winter and summer, but it is not obvious in the level of climate comfort. (4) A large area of natural lakes can improve the climate comfort in summer, but have no effect in winter. (5) In winter and summer, altitude has the greatest influence on climate comfort level, followed by latitude.