Abstract: In view of the potential threats to biodiversity in the process of rural revitalization and the needs of conservation and management, this study organically combined the theories and methods of biodiversity with society, economy, population, etc., and built the early warning index system of biodiversity in rural ecological landscape based on the DPSIR (Driving Force-Pressure-State-Impact-Response) model. Based on the principles of scientificity, relevance, practicality and comparability, the index system reflects the impact of human interference, global change, major disasters, measures and inputs taken by human to maintain the biodiversity of rural ecological landscape, and other factors on rural biodiversity, including 12 factors and 25 indicators of the driving force, pressure, state, impact and response of biodiversity. The mean square deviation method was used to determine the weight value of each index, divide the evaluation level. And the comprehensive index method was used to warn the biodiversity of the rural ecological landscape. The rural biodiversity in this study comprehensively considered the “Ecology, Production, Living” functions of the countryside. Therefore, the species of wild animals and plants, crop species, and rural green plants were all included in the calculation of the biodiversity index. This research method has been applied in the Paifang Community of Nanjing to give early warning to the high risk area. This study provides a new idea and method for biodiversity protection and evaluation in the revitalization and planning of green livable countryside, and it is of great significance to build a beautiful countryside with green harmony.