Soil extracellular enzyme activities and the abundance of nitrogen-cycling functional genes responded more to N addition than P addition in an Inner Mongolian meadow steppe

Nitrogen (N) and phosphorus (P) availability in soils commonly limit belowground biological processes in terrestrial ecosystems. Soil extracellular enzyme activities (EEAs) and microbial functional groups play critical roles in soil biological processes and nutrient cycling, yet their response to nutrient addition are poorly understood. To address this issue, we applied six fertilization treatments composed of combinations of N (0, 1.55, 13.95 g N m−2 yr−1) and P (0, 5.24 g P m−2 yr−1) for two years in a meadow steppe of Inner Mongolia. Soils were collected from each plot in July and August and analyzed for abundances of N-cycling genes and EEAs, and their relationships with treatments. The addition of N significantly increased C-acquisition enzyme activity and enzyme C:N and C:P ratios. Enzymatic stoichiometry indicated that N addition alleviated microbial demand for N, while it increased microbial C limitation. Microbial C and N limitation were significantly correlated with NH4+–N in July, yet they were correlated with soil water content (SWC) in August. The abundance of amoA significantly increased with N addition and was positively related to mineral-N accumulation. The abundance of denitrifier genes and gaseous N loss potential were accelerated by N addition in July, while a neutral effect was observed in August. Nitrate leaching potential was significantly increased by N addition, yet it declined with P addition in July. P addition also suppressed amoA abundance of ammonia oxidizing bacteria. Partial least squares path modelling indicated that N addition positively affected microbial-C limitation, soil N-loss potential and negatively affected microbial-N limitation. P addition negatively affected soil N-loss potential. Ultimately, this study highlights the importance of soil N availability in regulating microbial metabolism and soil N-loss potential, and enhances our understanding of the mechanisms responsible for variation in microbial nutrient cycling in meadow steppe soils. [Display omitted] •Soil extracellular enzyme activities (EEAs) positively responding to N addition•N addition enhanced microbial-C limitation, yet alleviated microbial N limitation.•The abundances of nitrifiers and denitrifiers significantly increased with N addition.•Nitrate leaching was increased by N addition while declined by P addition.•Altering of soil EEAs and N-cycle gene abundances mainly linked to soil N availability