The core of constructing regional ecological compensation mechanism is to define the subject and object of regional ecological compensation in a reasonable manner and to prioritize ecological compensation. This paper defined the subject and object of ecological compensation, and determined the level and order of ecological compensation areas and compensation payment from the perspective of the main functional areas by measuring the value of ecosystem services in Hubei Province. The results show that:(1) the spatial distribution of the total value of service, market value and non-market value of ecosystem services in Hubei Province is similar, showing a pattern of "high value in the West, low value in the middle and the East". The high-value areas are mostly concentrated in the key ecological function places and major production regions, while the low-value areas are mostly focused on the key development areas. (2) The ecological compensation of 30 key ecological functional areas is generally divided into five levels, showing a spatial distribution pattern of "high in the west and low in the east" with Fangxian topping the list. (3) The priority of ecological compensation in 29 major production regions is categorized into five levels with Suixian regarded as the first level region of ecological compensation. (4) The compensation payment level of 30 key development areas is divided into five levels, showing a spatial distribution pattern of "east first, west second". In carrying out ecological compensation payment, the key development areas at the national level should be prior to the key development areas at the provincial level.