Currently the potential of biomass crops, including grasses, is limited because most species have not been bred for this purpose. However traits such as lignification, phenolic cross-linking and carbohydrate accessibility, which are also important for nutritive quality in forage grasses, can affect potential biofuel quality in applications such as combustion, fast-pyrolysis or fermentation. A collection of Lolium and Festuca species known to exhibit a range of lignin, cell wall phenolic and carbohydrate concentrations have been used to test optimum characteristics for biofuel processing. This collection formed a “calibration” set for subsequent high through-put FTIR chemical screening of additional plant lines: (1) A set of Lolium-Festuca substitution lines, in which L. perenne chromosomes or chromosome segments are substituted by homoeologous regions of F. pratensis, that provide the potential to physically map biofuel traits to an individual chromosome or chromosome segment; (2) A maize transposon (Robertson’s Mutator) induced mutant collection, which provides the potential to identify gene sequences underlying important biochemical traits linked to biofuel as determined by FTIR analysis