Spatio-temporal dynamics and stakeholder engagement are critical components in the assessment of ecosystem services to support well-targeted and localised strategies for sustainable ecosystem management. Here, we investigated the recent changing pattern of ecosystem services’ value (ESV) in the Yangtze River Middle Reaches Megalopolis (YRMRM) - a region of vital economic and ecological importance in central China. Local experts’ opinions on the regional importance of different ecosystem services were acquired from an extensive survey and integrated into the localisation of ESV. We explored the shifting extents of the hot and cold spots of ESV and whether the ESV of a place could be related to its geographical location and distance. Our results showed a downtrend of ESV from 2000 to 2015, resulting from a loss in cultivated land and thereby a reduced ESV of the food supply. Cold spots intensified while hot spots diminished across the YRMRM. The ESVs of a place were distance-decay functions of the distances (d) between the place to the nearest provincial centre, lake and the Yangtze River. The maximum ESV was usually located at 42 km away from nearest provincial centre, 10 km away from the Yangtze River, and near the lake. This study adds important first-hand empirical data to the field of the stakeholder-driven valuation of localised ESV. Our results based on a comprehensive spatiotemporal assessment of ESV is conducive to promoting the coordination of sustainable development and ecological protection in YRMRM.