The agriculture sector is in a growing need to develop greenhouse gas (GHG) mitigation techniques to reduce the enhanced greenhouse effect. The challenge to the sector is not only to reduce net emissions but also increase production to meet growing demands for food, fiber, and biofuel. This study focuses on the changes in the GHG balance of three biofuel feedstock (biofuel sugarcane, energy-cane and sweet sorghum) considering changes caused by the adoption of conservationist practices such as reduced tillage, use of controlled-release fertilizers or when cultivation areas are converted from burned harvest to green harvest. Based on the Intergovernmental Panel on Climate Change (IPCC) (2006) balance and the Tools for the Reduction and Assessment of Chemical and Other Environmental Impacts (TRACI) characterization factors published by the EPA, the annual emission balance includes use energy (diesel and electricity), equipment, and ancillary materials, according to the mean annual consumption of supplies per hectare. The total amounts of GWP were 2740, 1791, and 1910 kg CO2e ha-1 y-1 for biofuel sugarcane, energy-cane and sweet sorghum, respectively, when produced with conventional tillage and sugarcane was burned prior to harvesting. Applying reduced tillage practices, the GHG emissions reduced to 13% for biofuel sugarcane, 23% for energy-cane and 8% for sweet sorghum. A similar decrease occurs when a controlled-release fertilizer practice is adopted, which helps reduce the total emission balance in 5%, 12% and 19% for biofuel sugarcane, energy-cane and sweet sorghum, respectively and a 31% average reduction in eutrophication potential. Moreover, the GHG emissions for biofuel sugarcane, with the adoption of green harvest, would result in a smaller GHG balance of 1924 kg CO2e ha-1 y-1, providing an effect strategy for GHG mitigation while still providing a profitable yield in Florida.