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Bioenergy developed from energy plants will play a more and more important role in future energy supply. Much attention has been paid to energy plants in recent years. As China has fairly limited cultivated land resources, the bioenergy development may mainly rely on the exploitation of marginal land. This study focused on the assessment of marginal land resources and bio-fuel potential in China using newly acquired data and Geographic Information System (GIS) techniques. A multi-factor analysis method was adopted to identify marginal lands for bioenergy development in China, with data of several main types of energy plants on the eco-environmental requirements and natural habits employed. A combined planting zonation strategy was proposed, which was targeted for five species of energy plants including Helianthus tuberous L., Pistacia chinensis, Jatropha curcas L., Cassava and Vernicia fordii. The results indicated that total area of marginal land exploitable for development of energy plants on a large scale was about 43.75 million ha. If 10% of this marginal land was fully utilized for growing the energy plants, the production of bio-fuel would be 13.39 million tons.