Abstract: We put forward the formulation and implementation of "ecological protection red line, environmental quality baseline, resource utilization upper limit and ecological environmental access list" guideline (also known as the “Three lines and one list” guideline) for comprehensive watershed planning, guided by the goal of environmental protection in the new era. Taking the Minjiang River basin as example, we put forward the principles, main content, indexes, methodology and implementation for the formulation of the guidelines. The “red line” is a spatial layout set up according to the major environmental problems and development needs, which is based on the division scheme of ecological protection “red line” issued by the local authorities. Key indexes of “environmental quality baseline” include target rate achieved for major indexes in water functional area, and water environment control objectives for key control sections. Key control indexes of “resource utilization upper limit” include total water consumption control indexes of the basin, and ecological base flow. Ecological environmental access list is essential for achieving the “Three- line” goal. The results show that the guideline runs through the entire process of comprehensive watershed planning, which can effectively coordinate the relationship between watershed development and the bottom line, and help ensure the ecological security of the basin.