[Objective] Studying the spatial-temporal variation and the influencing factors of ecosystem water source supply service in Upper Hanjiang Basin during 2000 and 2010, in order to provide basis for scientific management and policy formulation of regional water resources.[Methods] The water yield module within InVEST model was used to analyze the supply service.[Results] Forestland, wetland and construction land areas increased in the decade, while farmland area decreased. Grassland area remained stable. Annual mean vegetation coverage was in an overall rise with a gradient of 0.005 1/a. Water yields of upper Hanjiang basin in 2000, 2005 and 2010 were 46.70, 52.21 and 47.29 billion cubic meters, respectively.[Conclusion] Spatiotemporal variation of water source supply amount in the study area was a combination result of climate and land use change. Increased precipitation had a positive role in increasing water yield. Change of underlying surface caused by land use change and ecosystem improvement could be the main driving forces of the declined water supply in upper Hanjiang basin.