Illinois bundleflower (Desmanthus illinoensis (Michx.) Macmillan) has potential as a grain and forage legume for the American Midwest. Inoculant-quality rhizobia for this legume have been identified but not previously characterized. Rhizobia trapped from 20 soils in the natural range of the Illinois bundleflower had characteristics that placed them overwhelmingly within the species Rhizobium giardinii, one of the few occasions this species has been recovered from legumes, raising questions on the biogeography and spread of midwestern prairie rhizobia.