For watersheds with different land use needs under different development goal orientations, scientific and reasonable regulation of land use changes is an important cornerstone to achieve efficient land resource utilization and coordinated ecological-social-economic development in watersheds. Taking the Rao River Basin as an example, based on the Markov-FLUS coupled model with comprehensive consideration of both natural and anthropogenic aspects, 12 types of driving factors are selected, with 2005 land use data as the initial value, and actual land use type data from 2010 and 2015 are used to validate and correct the model, combined with scenario settings such as inertia development, cultivated land protection, and ecological priority. The simulation analysis of land use in the 2035 Rao River Basin is carried out to meet the optimal spatial allocation of national land under different development goal orientations of the basin. The results show that:(1) The model has strong applicability in the Rao River Basin, with an overall accuracy of 98.01% and the Kappa coefficient of 0.9627, which is greater than 0.80, providing a methodological reference for future land use simulations in the region. (2) The three scenario settings basically meet the different development demands of the Rao River Basin, and the land use structure of the basin changes significantly for different development goals, with the most significant changes in cultivated land and construction land under each scenario. It is mainly concentrated in Changjiang District, Zhushan District, Fuliang County, Leping City, Dexing City, and scattered in Poyang County and Wannian County. Under the inertia development scenario, construction land expands rapidly, with an increase of nearly 40%; under the cultivated land protection scenario, cultivated land shows the only positive growth, but the increase is only 1%; under the ecological priority scenario, forest land, grassland, and water area show increases at the same time, with 0.03%, 0.4%, and 1%, respectively. (3) Under the inertia development scenario, the cultivated land protection scenario and the ecological priority scenario, the trend of construction land expansion is 39%, 16%, and 12%, respectively, indicating that the implementation of the cultivated land protection and ecological priority policies needs to strengthen constraints on construction land expansion and further adjust the land use structure to improve land use efficiency. In summary, multi-scenario simulation of land use in Rao River Basin based on the Markov-FLUS model can provide basic support and multiple perspectives for future territorial spatial planning and socio-economic development decisions, and is of great significance for the efficient use of land resources and ecological environmental protection.