Substitution of soy-based biodiesel fuels for petroleum diesel will alter life cycle emissions for construction vehicles. A life cycle inventory was used to estimate fuel cycle energy consumption and emissions of selected pollutants and greenhouse gases. Real-world measurements using a portable emission measurement system (PEMS) were made forfive backhoes, four front-end loaders, and six motor graders on both fuels from which fuel consumption and tailpipe emission factors of CO, HC, NO(x), and PM were estimated. Life cycle fossil energy reductions are estimated it 9% for B20 and 42% for B100 versus petroleum diesel based on the current national energy mix. Fuel cycle emissions will contribute a larger share of total life cycle emissions as new engines enter the in-use fleet. The average differences in life cycle emissions for B20 versus diesel are: 3.5% higher for NO(x); 11.8% lower for PM, 1.6% higher for HC, and 4.1% lower for CO. Local urban tailpipe emissions are estimated to be 24% lower for HC, 20% lower for CO, 17% lower for PM, and 0.9% lower for NO(x). Thus, there are environmental trade-offs such as for rural vs urban areas. The key sources of uncertainty in the B20 LCI are vehicle emission factors.