Acidification Dominates over Hypoxia in Controlling Estuarine Nitrogen Removal Dynamics under Coupled Stressors

As critical transitional zones between land and sea, estuaries are confronting the dual threats of increasing acidification and hypoxia driven by human activities and climate change. However, the combined effects of these stressors on estuarine nitrogen removal processes remain poorly understood. In this study, using stable-isotope tracing and molecular techniques in the Yangtze estuary, we found that hypoxia promoted N removal, yet concurrent acidification can override this effect, leading to net inhibition and a consequent reduction in estuarine nitrogen removal capacity. However, in seasonally hypoxic zones, these combined stressors generally enhanced nitrogen removal rates (by up to 34.4%), which suggests a degree of resilience under such perturbations. Nevertheless, the concurrent acidification-hypoxia in seasonally hypoxic areas stimulated N₂O emissions (8.5-44.4%), which may intensify climate forcing and thereby further exacerbate these environmental stressors. Metagenomic and quantitative PCR analyses corroborated these response patterns, revealing coordinated changes in the abundance and expression of key nitrogen-removal genes, as well as divergent microbial response strategies and niche differentiation under acidification-hypoxia stress. This study elucidates the previously overlooked interactive effects of acidification and hypoxia on estuarine nitrogen removal, providing a mechanistic basis for refining biogeochemical models to improve the reliability of simulations under multiple stressors.