[Objective] Taking the urban agglomeration of Central Yunnan Province in the plateau karst area as an example, the terrain gradient was introduced to explore the impact of land use/cover change (LUCC) on ecosystem service value (ESV) in the process of rapid urbanization, in order to provide theoretical guidance for the improvement of ecosystem service value and land use planning of other similar areas. [Methods] Based on the land use data from 2000 to 2020, combined with the topographic index, modified ESV equivalent and hotspot analysis method, the spatial and temporal variation characteristics of LUCC and ESV on the terrain gradient the study area from 2000 to 2020 was quantitatively discussed, and the impact of LUCC on ESV was analyzed through the ESV profit and loss flow table. [Results] ① From 2000 to 2020, the cultivated land and forest land in the study area decreased by 1.21×10⁵ hm² and 1.19×10⁵ hm², respectively, while the construction land increased by 2.16×10⁵ hm². Cultivated land, water land and construction land were mainly concentrated on low terrain gradient, and forest land was mainly concentrated on high terrain gradient. ② The value of ESV decreased from 119.64 billion yuan to 113.75 billion yuan from 2000 to 2020, with a total decrease of 5.89 billion yuan. ESV had significant spatial differentiation characteristics on the terrain gradient. ESV high-value areas were mainly distributed in the western and northern regions with high altitudes and steep slopes, while ESV low-value areas were on the contrary. ③ The loss of forest land and water land and the occupation of construction land on cultivated land led to heavy loss of ESV. The loss areas were mainly distributed in urban areas in the central part of Kunming City and the northern part of Honghe Prefecture. From 2010 to 2020, due to the implementation of the policy of returning cultivated land to forest land, grassland, and water land, the area of forest land and water land in the northern part of Honghe Prefecture and the northern part of Qujing City increased, and the ESV value-added areas appeared. [Conclusion] During the study period, LUCC had a great impact on ESV and showed obvious spatial differences on the terrain gradient. Measures such as improving and protecting the existing forests, grasslands and lakeside wetlands, and strict control of ESV loss cold spots can effectively improve the ecosystem service value and promote regional sustainable land use of the study area.