Abstract: A demonstration area for European Union-China Biodiversity Programme (ECBP) in Libo, Guizhou, China, as a representative karst area, was involved in this study. Firstly, the evaluation of ecological sensitivity was applied to regionalization for formulating the land consolidation strategies, including land consolidation zone, conservation zone of natural vegetation, conservation zone of forest edge, reconstruction zone of river ecosystem, and ecological corridor area. Secondly, by the ‘least-cost’ modeling, the ecological corridors for enhancing the connectivity for protective animals were chosed. The hot spots in landscape were defined by the importance in ecosystem to be protected as habitat patches. Following the optimization of landscape pattern, ecological engineering including irrigation and drain engineering, restoration of land productive, vegetation reconstruction, and road layout were designed for biodiversity conservation. The results showed that the interdependence of cultural landscapes, landscape pattern and ecological infrastructure optimization in land consolidation project planning could coordinate the social, economic, and ecological benefits. This study can supply a theoretical reference and practical approach for biodiversity conservation in land consolidation planning.