Sharon A. Billings

Nitrogen (N) additions often decrease soil respiration and increase soil organic carbon (C) stock. However, it is unclear how microbial substrates may shift with N additions and increasing temperature. Leveraging 12 years of N fertilization experiments and the associated shift in the dominant vegetation from C 4 to C 3 , we explored the δ 13 C‐CO 2 and temperature sensitivities of respired CO 2 and extracellular enzyme activities in control and fertilized soils. N additions increased cellulose‐decaying extracellular enzyme activity while respiration remained similar between the control and fertilized soils. Temperature sensitivity of cellulose‐decaying extracellular enzyme activity decreased with the N additions. The δ 13 C‐CO 2 data reveal that, as temperature increased, microbes in fertilized soils changed their dominant substrate from bulk soil organic C to plant litterfall. Our results suggest that long‐term N fertilization imposed C limitation on microbes, leading to enhanced microbial efforts to acquire C. This study highlights how long‐term N additions can promote the relative preservation of organic C in mineral soil while litterfall, the precursor to mineral‐associated C, is increasingly decayed as temperatures increase.