The root lesion nematode, Pratylenchus neglectus, is one of the most damaging nematodes to affect wheat worldwide. The nematode is widely distributed in Montana, primarily affecting winter wheat within the state. Managing the nematode primarily involves rotation to resistant and moderately resistant crops (peas, lentils, and barley). A nematode survey was conducted across the state nearly 10 years after an initial survey, to reassess the nematode threat and assess the impact of changing trends in crop rotations. To assess the broad applicability of rotation crops to control P. neglectus across Montana, greenhouse trials were conducted to challenge rotational crops using eight populations of P. neglectus collected from geographically diverse locations across the state. In the trials, conducted with four Montana crops, a significant interaction was detected between crop and nematode population (analysis of variance P < 0.001). Populations from Hill, Dawson, and Chouteau counties were found to be pathogenic on barley. Male nematodes were detected in seven of the eight pot culture populations, and these were confirmed to be P. neglectus by morphological and molecular methods. These results suggest a re-evaluation of barley and lentils as a management option for P. neglectus in Montana, as pathotypes for each exist within the state.