Abstract: As one of the major food production areas in China, the efficiency and mode of land resource utilization in Huaihe River Basin is directly related to the development of regional agriculture and urban economy, so the research on the carrying capacity of land resources in Huaihe River Basin and its influencing factors has very important theoretical value and practical significance. In this paper, a comprehensive evaluation index system was constructed from the three dimensions of carrying strength, carrying efficiency and carrying potential, and based on the entropy weight TOPSIS model and the spatial autocorrelation model, the land resource carrying capacity levels of 35 prefectural-level cities in the Huaihe River Basin were measured in 2006—2021, and the spatial and temporal variations of the land resource carrying capacity were analyzed empirically. The results showed that, during the study period, the carrying capacity of land resources in all prefectural-level cities in the Huaihe River Basin showed an upward trend, with a high spatial distribution pattern in the east and a low spatial distribution pattern in the west, a positive spatial autocorrelation in the whole, and a spatial differentiation pattern of “small agglomeration and large dispersion” in the local area.