Symbiotic dinitrogen fixation is seasonal and strongly regulated in water-limited environments

Plants, especially perennials, growing in drylands and seasonally dry ecosystems are uniquely adapted to dry conditions. Legume shrubs and trees, capable of symbiotic dinitrogen (N₂ ) fixation, often dominate in drylands. However, the strategies that allow symbiotic fixation in these ecosystems, and their influence on the nitrogen cycle, are largely unresolved. We evaluated the climatic, biogeochemical and ontogenetic factors influencing nitrogen fixation in an abundant Mediterranean legume shrub, Calicotome villosa. We measured nodulation, fixation rate, nitrogen allocation and soil biogeochemistry in three field sites over a full year. A controlled experiment evaluated differences in plant regulation of fixation as a function of soil nutrient availability and seedling and adult developmental stages. We found a strong seasonal pattern, shifting between high fixation rates during the rainy season at flowering and seed-set times to almost none in the rainless season. Under controlled conditions, plants downregulated fixation in response to soil nitrogen availability, but this response was stronger in seedlings than in adult shrubs. Finally, we did not find elevated soil nitrogen under N₂ -fixing shrubs. We conclude that seasonal nitrogen fixation, regulation of fixation, and nitrogen conservation are key adaptations influencing the dominance of dryland legumes in the community, with broader consequences on the ecosystem nitrogen cycle.