The growing awareness that plants might use a variety of nitrogen (N) forms, both organic and inorganic, has raised questions about the role of resource partitioning in plant communities. It has been proposed that coexisting plant species might be able to partition a limited N pool, thereby avoiding competition for resources, through the uptake of different chemical forms of N. In this study, we used in situ stable isotope labeling techniques to assess whether coexisting plant species of a temperate grassland (England, UK) display preferences for different chemical forms of N, including inorganic N and a range of amino acids of varying complexity. We also tested whether plants and soil microbes differ in their preference for different N forms, thereby relaxing competition for this limiting resource. We examined preferential uptake of a range of 13C15N-labeled amino acids (glycine, serine, and phenylalanine) and 15N-labeled inorganic N by coexisting grass species and soil microbes in the field. Our data show that while coexisting plant species simultaneously take up a variety of N forms, including inorganic N and amino acids, they all showed a preference for inorganic N over organic N and for simple over the more complex amino acids. Soil microbes outcompeted plants for added N after 50 hours, but in the long term (33 days) the proportion of added 15N contained in the plant pool increased for all N forms except for phenylalanine, while the proportion in the microbial biomass declined relative to the first harvest. These findings suggest that in the longer term plants become more effective competitors for added 15N. This might be due to microbial turnover releasing 15N back into the plant–soil system or to the mineralization and subsequent plant uptake of 15N transferred initially to the organic matter pool. We found no evidence that soil microbes preferentially utilize any of the N forms added, despite previous studies showing that microbial preferences for N forms vary over time. Our data suggest that coexisting plants can outcompete microbes for a variety of N forms, but that such plant species show similar preferences for inorganic over organic N.