The utilization of native grass species for potential biomass feedstocks of the emerging ethanol industry requires more information about their cellulose and hemicellulose concentration. Ten native species were grown at seven sites across the prairie region of western Canada for two to four growing seasons. Northern wheatgrass, Elymus lanceolatus, produced high concentrations of cellulose (363 g kg–1) but low concentrations of hemicellulose (266 g kg–1). Green needlegrass, Nasella viridula, produced high concentrations of both constituents (351 and 307 g kg–1). Four warm-season grasses, big bluestem, Andropogon gerardii, little bluestem, Schizachyrium scoparium, prairie sandreed, Calamovilfa longifolia, and switchgrass, Panicum virgatum, had 346, 342, 340 and 338 g kg–1, respectively, concentrations of cellulose and also exhibited a positive response to temperature that resulted in increased hemicellulose concentration. Accumulated thermal time (degree day base 10°C) was correlated to hemicellulose concentrations in the warm-season grasses but not for cool-season grasses. Holocellulose (cellulose + hemicellulose) concentration differences varied among site-years but warmseason grasses were more stable in hollocellulose concentration than cool-season grasses.