Native grasslands that support diverse populations of birds are being converted to cropland at an increasing rate in the Prairie Pothole Region of North America. Although limited funding is currently available to mitigate losses, accurate predictions of probability of conversion would increase the efficiency of conservation measures. We studied conversion of native grassland to cropland in the Missouri Coteau region of North and South Dakota (U.S.A.) during 1989-2003. We estimated the probability of conversion of native grassland to cropland with satellite imagery and logistic regression models that predicted risk of conversion and by comparing the overlap between areas of high biological value and areas most vulnerable to conversion. Annualized probability of conversion was 0.004, and 36,540 ha of native grassland were converted to cropland during the period of our study. Our predictive models fit the data and correctly predicted 70% of observed conversions of grassland. Probability of conversion varied spatially and was correlated with landscape features like amount of surrounding grassland, slope, and soil productivity. Tracts of high biological value were not always at high risk of conversion. We concluded the most biologically valuable areas that are most vulnerable to conversion should be prioritized for conservation. This approach can be applied broadly to other systems and offers great utility for implementing conservation in areas with spatially variable biological value and probability of conversion.