[Objective] The agglomeration characteristics and evolution trend of land use conflicts was scientifically identified to provide important reference for promoting the rational layout of territorial space and regional sustainable development. [Methods] The spatial-temporal evolution trend of land use conflict in the Wanjiang urban belt from 1990 to 2020 was quantitatively analyzed by constructing a land use conflict measurement model based on the landscape index. The distance weight matrix was determined by setting the optimal spatial threshold to explore the agglomeration effect of land use conflict more accurately. [Results] ① Significant changes occurred in the land use structure of the Wanjiang urban belt from 1990 to 2020. The area of cultivated and forest land decreased continuously, and the scale of construction land increased significantly. ② The distribution pattern of regional land use conflicts was “higher in the north and lower in the south,” with areas of weak and medium spatial conflicts occupying the main position. Serious conflict areas were concentrated in Hefei City, Wuhu City, Jin’an District of Lu’an City, and the coastal areas on both sides of the Yangtze River system. ③ The land use conflict index changed significantly in urban areas during the study period, especially in the east of Hefei City, the north of Wuhu City, and the southeast of Chuzhou City. Under the influence of urbanization, the conflict gravity center gradually shifted southward, and the serious conflict zone shifted from the northwest to the southeast. ④ 5 km was selected as the optimal distance analysis threshold. The high-high aggregation conflict zone was centered on Hefei City, while the low-low aggregation conflict zone showed no agglomeration phenomenon. [Conclusion] The land use conflict in Wanjiang urban belt has the potential to intensify, and the main conflict areas show a shifting trend. In the future, it is necessary to strengthen the rational distribution of regional land use and focus on the balance and stability between urbanization development and ecological protection.