Ecological networks have been considered to be an effective strategy to counter habitat fragmentation, which is occurring in the urbanization context. Researchers and planners have begun to integrate ecological networks in both thematic planning and urban or regional planning for biodiversity conservation and sustainable development. In this paper, we integrated the least-cost path model, network structure analysis, and gravity model to construct a regional ecological network of Poyang Lake Eco-economic Region, China. The least-cost model was improved by integrating three resistance factors, including land cover, slope, and human-induced pressure into it, presenting the landscape characteristics comprehensively in corridor simulation. The network structure analysis was employed to assess the proposed ecological network and the gravity model was used to identify the relative significance of network elements. Results indicated that the proposed hierarchical ecological network has good circuitry, complexity, and connectivity. Spatial analysis of prioritized network elements and breaking points revealed the regional connectivity condition, presenting significant reference to decision making in both land-use management and network implementation. Our study demonstrated that the integrated methodology is effective in regional ecological network planning and shows feasibility to be applied to other regions.