Phosphorus-driven regime shift from heterotrophic to autotrophic diazotrophs in a deep alpine lake

Biological nitrogen fixation plays a critical role in maintaining primary production, particularly in systematic nitrogen deficiency. However, little is known about the dynamics within diazotrophic community facing ongoing nutrient enrichment in freshwater lakes. Here, a consecutive five-year investigation on diazotrophic community was conducted in Lake Fuxian, an oligotrophic deep alpine lake on the trajectory to eutrophic state. Results showed a regime shift from heterotrophic to autotrophic diazotrophs induced by total phosphorus (TP) enrichment. Specifically, heterotrophic diazotrophs dominated the diazotrophic community when TP was lower than 21.8 μg/L, whereas heterotrophic diazotrophs or diazotrophic Cyanobacteria randomly dominated when TP ranged between 21.8 μg/L and 28.8 μg/L. When TP was higher than 28.8 μg/L, diazotrophic Cyanobacteria accounted for 60.4%-97.7% of the total N₂-fixers, indicating diazotrophic biodiversity significantly declined under TP enrichment scenario. Moreover, the dominance of diazotrophic Cyanobacteria further facilitated phytoplankton growth, which strengthened positive feedback between phytoplankton and phosphorus under nitrogen deficiency conditions. This is the first report on the threshold-like state responses of freshwater diazotrophs to environmental drivers. Our study expands the knowledge of the diazotrophic dynamics in freshwater ecosystems and contributes quantitative evidence of ecological thresholds for future environmental policymaking.