Marginal agricultural land is estimated for biofuel production in Africa, China, Europe, India, South America, and the continental United States, which have major agricultural production capacities. These countries/regions can have 320?702 million hectares of land available if only abandoned and degraded cropland and mixed crop and vegetation land, which are usually of low quality, are accounted. If grassland, savanna, and shrubland with marginal productivity are considered for planting low-input high-diversity (LIHD) mixtures of native perennials as energy crops, the total land availability can increase from 1107?1411 million hectares, depending on if the pasture land is discounted. Planting the second generation of biofuel feedstocks on abandoned and degraded cropland and LIHD perennials on grassland with marginal productivity may fulfill 26?55% of the current world liquid fuel consumption, without affecting the use of land with regular productivity for conventional crops and without affecting the current pasture land. Under the various land use scenarios, Africa may have more than one-third, and Africa and Brazil, together, may have more than half of the total land available for biofuel production. These estimations are based on physical conditions such as soil productivity, land slope, and climate.