Nitrogen (N) saturation theory proposes that an ecosystem might switch from N limitation to carbon (C), phosphorus (P), or other nutrient limitations if it receives continuous N input. Yet, after N limitation is removed, which nutrient is the most limited and whether topography modulates such change is rarely tested at a microbial level. Here, we conducted a two-year N addition experiment under two different topography positions (i.e. a slope and a valley) in a N-saturated subtropical forest. Soil enzyme activity was measured, and ecoenzymatic stoichiometry indexes were calculated as indicators of microbial resource limitation.

In the valley, two-year N addition changed the activity of all studied enzymes to various degrees. As a result, microbial C limitation was aggravated in the valley, and consequently microbial decomposition of soil labile organic C increased, but microbial P limitation was alleviated due to the stoichiometry balance. On the slope, however, N addition did not significantly change the activity of the studied enzymes, and did not alter the status of microbial resource limitation.

These results indicate that C is a more limited element for microbial growth than P after removing N limitation, but we also highlight that topography can regulate the effect of N deposition on soil microbial resource limitation in subtropical forests. These findings provide useful supplements to the N saturation theory.