Loss of native grasslands by anthropogenic disturbances has reduced availability and connectivity of habitat for many grassland species. A primary threat to contiguous grasslands is the encroachment of woody vegetation, which is spurred by disturbances that take on many forms from energy development, fire suppression, and grazing. These disturbances are exacerbated by natural- and human-driven cycles of changes in climate punctuated by drought and desertification conditions. Encroachment of honey mesquite (Prosopis glandulosa) into the prairies of southeastern New Mexico has potentially limited habitat for numerous grassland species, including lesser prairie-chickens (Tympanuchus pallidicinctus). To determine the magnitude of impacts of distribution of mesquite and how lesser prairie-chickens respond to mesquite presence on the landscape in southeastern New Mexico, we evaluated seasonal space use of lesser prairie-chickens in the breeding and nonbreeding seasons. We derived several remotely sensed spatial metrics to characterize the distribution of mesquite. We then used these data to create population-level resource utilization functions and predict intensity of use of lesser prairie-chickens across our study area. Home ranges were smaller in the breeding season compared with the nonbreeding season; however, habitat use was similar across seasons. During both seasons, lesser prairie-chickens used areas closer to leks and largely avoided areas with mesquite. Relative to the breeding season, during the nonbreeding season habitat use suggested a marginal increase in mesquite within areas of low intensity of use, yet aversion to mesquite was strong in areas of medium to high intensity of use. To our knowledge, our study is the first to demonstrate a negative behavioral response by lesser prairie-chickens to woody encroachment in native grasslands. To mitigate one of the possible limiting factors for lesser prairie-chickens, we suggest future conservation strategies be employed by land managersto reduce mesquite abundance in the southern portion of their current range.