Highlights What are the main findings? The LS and C factors of the RUSLE model were locally adjusted. Soil conservation service in the Guizhou Plateau exhibited an improving trend from 2000 to 2020. What are the implications of the main findings? The relationship of ecological services exhibited significant spatial variation. Ecological engineering improved the synergy of ecological services in rocky desertification areas.Highlights What are the main findings? The LS and C factors of the RUSLE model were locally adjusted. Soil conservation service in the Guizhou Plateau exhibited an improving trend from 2000 to 2020. What are the implications of the main findings? The relationship of ecological services exhibited significant spatial variation. Ecological engineering improved the synergy of ecological services in rocky desertification areas.Abstract The Guizhou Plateau, with the most extensive and representative karst landforms worldwide, is characterized by severe soil erosion and a highly fragile ecological environment. However, large-scale assessments of soil conservation services in this region remain limited. A key challenge lies in identifying appropriate datasets and methodologies for regional-scale soil conservation service evaluations, particularly under conditions of data scarcity or limited data accuracy. In this study, Unmanned Aerial Vehicle imagery, runoff plot observations, ground survey data, and multi-source satellite remote sensing data were integrated to refine LS and C in the Revised Universal Soil Loss Equation (RUSLE), thereby establishing a parameterized and localized soil erosion model. This improvement provided a methodological foundation for soil conservation service research in the region. Subsequently, the spatiotemporal variations in soil conservation services in the Guizhou Plateau over the past two decades were assessed. Furthermore, the relationships between soil conservation services and other key ecosystem services, including water conservation and carbon sequestration, were quantitatively examined, and the driving factors were analyzed. Soil conservation on the Guizhou Plateau exhibited an improving trend from 2000 to 2020. In karst areas, the relationship between soil conservation and water conservation was primarily influenced by temperature, altitude, and vegetation coverage, whereas in non-karst areas, it was regulated by rainfall and slope. Ecological restoration projects have enhanced the synergy between soil conservation and carbon sequestration by promoting vegetation cover. These findings could contribute to the next stage of ecological engineering initiatives and ecological policy implementation in Guizhou.