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Nitrogen deposition alleviates phosphorus-induced imbalances in soil enzyme stoichiometry


Abstract

Soil extracellular enzymes are critical drivers of carbon (C) and nutrient cycling in terrestrial ecosystems. However, the effects of phosphorus (P) additions on soil enzyme activities and stoichiometries, particularly under varying nitrogen (N) addition regimes worldwide, are not well comprehended. Here, we conducted a meta-analysis based on 155 field studies across various ecosystems (forests, croplands and grasslands), which shows that P enrichment conditions enhances C-acquiring enzymes but has no effect on the enzymatic C:N ratio. P addition alone reduces P-acquiring enzymes by 14% without affecting N-acquiring enzymes. In contrast, P combined with N addition boosts N-acquiring enzymes by 21% while leaving P-acquiring enzymes unchanged. Notably, the combined effect of P and N addition on microbial C limitation (assessed via vector length) and enzymatic stoichiometries is less pronounced than that of P addition alone. Key drivers of these responses include mean annual precipitation, soil microbial biomass, and its stoichiometries. These results suggest that N addition mitigates the stoichiometric imbalance and microbial C limitation induced by P addition, potentially promoting soil organic C accumulation. Our findings emphasize the critical need to account for such interactive effects in models predicting soil biogeochemical cycles under future changes in global exogenous N and P inputs.

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The data used can be found in Figshare (https://doi.org/10.6084/m9.figshare.30737228).

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R code used can be found in Figshare (https://doi.org/10.6084/m9.figshare.30370738).

References

  1. Hou, E. et al. Global meta-analysis shows pervasive phosphorus limitation of aboveground plant production in natural terrestrial ecosystems. Nat. Commun. 11, 637 (2020).

    Google Scholar 

  2. Mahowald, N. et al. Global distribution of atmospheric phosphorus sources, concentrations and deposition rates, and anthropogenic impacts. Glob. Biogeochem. Cycle. 22, GB4026 (2008).

  3. Penuelas, J. et al. Human-induced nitrogen-phosphorus imbalances alter natural and managed ecosystems across the globe. Nat. Commun. 4, 2934 (2013).

    Google Scholar 

  4. Margalef, O. et al. The effect of global change on soil phosphatase activity. Glob. Change Biol. 27, 5989–6003 (2021).

    Google Scholar 

  5. Wu, W. et al. Meta-analysis of the impacts of phosphorus addition on soil microbes. Agric. Ecosyst. Environ. 340, 108180 (2022).

    Google Scholar 

  6. Sun, Y. et al. Phosphorus additions imbalance terrestrial ecosystem C:N:P stoichiometry. Glob. Change Biol. 28, 7353–7365 (2022).

    Google Scholar 

  7. Li, J. et al. Soil enzyme activity and stoichiometry in response to precipitation changes in terrestrial ecosystems. Soil Biol. Biochem. 191, 109321 (2024).

    Google Scholar 

  8. Sinsabaugh, R. L. et al. Stoichiometry of soil enzyme activity at global scale. Ecol. Lett. 11, 1252–1264 (2008).

    Google Scholar 

  9. Fatemi, F. R., Fernandez, I. J., Simon, K. S. & Dail, D. B. Nitrogen and phosphorus regulation of soil enzyme activities in acid forest soils. Soil Biol. Biochem. 98, 171–179 (2016).

    Google Scholar 

  10. Sinsabaugh, R. L., Hill, B. H. & Shah, J. J. F. Ecoenzymatic stoichiometry of microbial organic nutrient acquisition in soil and sediment. Nature 462, 795–U117 (2009).

    Google Scholar 

  11. He, P., Zhang, Y., Shen, Q., Ling, N. & Nan, Z. Microbial carbon use efficiency in different ecosystems: a meta-analysis based on a biogeochemical equilibrium model. Glob. Change Biol. 29, 4758–4774 (2023).

    Google Scholar 

  12. Mori, T. Does ecoenzymatic stoichiometry really determine microbial nutrient limitations?. Soil Biol. Biochem. 146, 107816 (2020).

    Google Scholar 

  13. Rosinger, C., Rousk, J. & Sandén, H. Can enzymatic stoichiometry be used to determine growth-limiting nutrients for microorganisms? A critical assessment in two subtropical soils. Soil Biol. Biochem. 128, 115–126 (2019).

    Google Scholar 

  14. Mori, T., Rosinger, C. & Margenot, A. J. Enzymatic C:N:P stoichiometry: questionable assumptions and inconsistencies to infer soil microbial nutrient limitation. Geoderma 429, 116242 (2023).

    Google Scholar 

  15. Mori, T., Aoyagi, R., Kitayama, K. & Mo, J. Does the ratio of β-1,4-glucosidase to β-1,4-N-acetylglucosaminidase indicate the relative resource allocation of soil microbes to C and N acquisition?. Soil Biol. Biochem. 160, 108363 (2021).

    Google Scholar 

  16. Kunito, T. et al. Ecoenzymatic stoichiometry as a temporally integrated indicator of nutrient availability in soils. Soil Sci. Plant Nutr. 70, 246–269 (2024).

    Google Scholar 

  17. Liu, H. et al. Nitrogen addition induces microbial phosphorus limitations in bulk soil but not in rhizospheric soil: a global analysis. Soil Res. 252, 106609 (2025).

    Google Scholar 

  18. Cui, Y. et al. Ecoenzymatic stoichiometry reveals widespread soil phosphorus limitation to microbial metabolism across Chinese forests. Commun. Earth Environ. 3, 184 (2022).

    Google Scholar 

  19. Cui, Y. et al. Global patterns of nutrient limitation in soil microorganisms. Proc. Natl. Acad. Sci. USA 122, e2424552122 (2025).

    Google Scholar 

  20. Moorhead, D., Cui, Y., Sinsabaugh, R. & Schimel, J. Interpreting patterns of ecoenzymatic stoichiometry. Soil Biol. Biochem. 180, 108997 (2023).

    Google Scholar 

  21. Cui, Y., Moorhead, D. L., Peng, S., Sinsabaugh, R. L. & Peñuelas, J. Predicting microbial nutrient limitations from a stoichiometry-based threshold framework. Innov. Geosci. 2, 100048 (2024).

    Google Scholar 

  22. Allison, S. D. & Vitousek, P. M. Responses of extracellular enzymes to simple and complex nutrient inputs. Soil Biol. Biochem. 37, 937–944 (2005).

    Google Scholar 

  23. Zhou, Z. H., Wang, C. K. & Jin, Y. Stoichiometric responses of soil microflora to nutrient additions for two temperate forest soils. Biol. Fertil. Soils 53, 397–406 (2017).

    Google Scholar 

  24. Liu, M. H., Gan, B. P., Li, Q., Xiao, W. F. & Song, X. Z. Effects of nitrogen and phosphorus addition on soil extracellular enzyme activity and stoichiometry in Chinese fir (Cunninghamia lanceolata) forests. Front. Plant Sci. 13, 834184 (2022).

    Google Scholar 

  25. Wang, F. C. et al. Divergent responses of soil microbial community to long-term nitrogen and phosphorus additions in a subtropical Chinese fir plantation. Catena 242, 108132 (2024).

    Google Scholar 

  26. Keane, J. B. et al. Soil C, N and P cycling enzyme responses to nutrient limitation under elevated CO2. Biogeochemistry 151, 221–235 (2020).

    Google Scholar 

  27. Deng, H. Y. et al. Soil ecosystem multifunctionality is strongly linked with crop yield after four decades chemical fertilization in black soil. Agric. Ecosyst. Environ. 368, 109007 (2024).

    Google Scholar 

  28. Mori, T. The ratio of β−1,4-glucosidase activity to phosphomonoesterase activity remains low in phosphorus-fertilized tropical soils: a meta-analysis. Appl. Soil Ecol. 180, 104635 (2022).

    Google Scholar 

  29. Forrester, D. I. & Bauhus, J. A review of processes behind diversity-productivity relationships in forests. Curr. Rep. 2, 45–61 (2016).

    Google Scholar 

  30. Chen, X. & Chen, H. Y. H. Plant mixture balances terrestrial ecosystem C:N:P stoichiometry. Nat. Commun. 12, 4562 (2021).

    Google Scholar 

  31. Castle, S. C. et al. Nutrient limitation of soil microbial activity during the earliest stages of ecosystem development. Oecologia 185, 513–524 (2017).

    Google Scholar 

  32. DeForest, J. L., Dorkoski, R., Freedman, Z. B. & Smemo, K. A. Multi-year soil microbial and extracellular phosphorus enzyme response to lime and phosphate addition in temperate hardwood forests. Plant Soil 464, 391–404 (2021).

    Google Scholar 

  33. Ma, S. H. et al. Inconsistent responses of soil microbial community structure and enzyme activity to nitrogen and phosphorus additions in two tropical forests. Plant Soil 460, 453–468 (2021).

    Google Scholar 

  34. Dove, N. C., Torn, M. S., Hart, S. C. & Tas, N. Metabolic capabilities mute positive response to direct and indirect impacts of warming throughout the soil profile. Nat. Commun. 12, 2089 (2021).

    Google Scholar 

  35. Moorhead, D. L., Sinsabaugh, R. L., Hill, B. H. & Weintraub, M. N. Vector analysis of ecoenzyme activities reveal constraints on coupled C, N and P dynamics. Soil Biol. Biochem. 93, 1–7 (2016).

    Google Scholar 

  36. Zuccarini, P. et al. Effects of nitrogen deposition on soil enzymatic activity and soil microbial community in a Mediterranean holm oak forest. Geoderma 430, 116354 (2023).

    Google Scholar 

  37. Huang, X. et al. Nitrogen deposition mitigates long-term phosphorus input-induced stimulative effects on soil respiration in a tropical forest. Geoderma 453, 117142 (2025).

    Google Scholar 

  38. Yuan, X. et al. Linkages of stoichiometric imbalances to soil microbial respiration with increasing nitrogen addition: Evidence from a long-term grassland experiment. Soil Biol. Biochem. 138, 107580 (2019).

    Google Scholar 

  39. Luo, X., Zhang, L., Lin, Y., Wen, D. & Hou, E. Nitrogen availability mediates soil organic carbon cycling in response to phosphorus supply: a global meta-analysis. Soil Biol. Biochem. 185, 109158 (2023).

    Google Scholar 

  40. Jiang, J. et al. Antagonistic and additive interactions dominate the responses of belowground carbon-cycling processes to nitrogen and phosphorus additions. Soil Biol. Biochem. 156, 108216 (2021).

    Google Scholar 

  41. Zheng, X. et al. Interactions between nitrogen and phosphorus in modulating soil respiration: a meta-analysis. Sci. Total Environ. 905, 167346 (2023).

    Google Scholar 

  42. Yu, Q. et al. Differential responses of soil phosphorus fractions to nitrogen and phosphorus fertilization: a global meta-analysis. Glob. Biogeochem. Cycle 38, e2023GB008064 (2024).

    Google Scholar 

  43. Wang, J., Wu, Y., Li, J., He, Q. & Bing, H. Soil enzyme stoichiometry is tightly linked to microbial community composition in successional ecosystems after glacier retreat. Soil Biol. Biochem. 162, 108429 (2021).

    Google Scholar 

  44. Fang, X. et al. Phosphorus addition alters the response of soil organic carbon decomposition to nitrogen deposition in a subtropical forest. Soil Biol. Biochem. 133, 119–128 (2019).

    Google Scholar 

  45. Zhang, H. et al. Phosphorus addition stimulates overall carbon acquisition enzymes but suppresses overall phosphorus acquisition enzymes: a global meta-analysis. Agric. Ecosyst. Environ. 375, 109219 (2024).

    Google Scholar 

  46. Jing, X. et al. Neutral effect of nitrogen addition and negative effect of phosphorus addition on topsoil extracellular enzymatic activities in an alpine grassland ecosystem. Appl. Soil Ecol. 107, 205–213 (2016).

    Google Scholar 

  47. Xiao, W., Chen, X., Jing, X. & Zhu, B. A meta-analysis of soil extracellular enzyme activities in response to global change. Soil Biol. Biochem. 123, 21–32 (2018).

    Google Scholar 

  48. Turner, B. L. & Wright, S. J. The response of microbial biomass and hydrolytic enzymes to a decade of nitrogen, phosphorus, and potassium addition in a lowland tropical rain forest. Biogeochemistry 117, 115–130 (2014).

    Google Scholar 

  49. Prescott, C. E. et al. Surplus carbon drives allocation and plant-soil interactions. Trends Ecol. Evol. 35, 1110–1118 (2020).

    Google Scholar 

  50. Mouginot, C. et al. Elemental stoichiometry of fungi and bacteria strains from grassland leaf litter. Soil Biol. Biochem. 76, 278–285 (2014).

    Google Scholar 

  51. Gong, J. R. et al. Phosphorus fertilization affects litter quality and enzyme activity in a semiarid grassland. Plant Soil 492, 91–108 (2023).

    Google Scholar 

  52. Nannipieri, P., Trasar-Cepeda, C. & Dick, R. P. Soil enzyme activity: a brief history and biochemistry as a basis for appropriate interpretations and meta-analysis. Biol. Fertil. Soils 54, 11–19 (2018).

    Google Scholar 

  53. Puissant, J. et al. The pH optimum of soil exoenzymes adapt to long term changes in soil pH. Soil Biol. Biochem. 138, 107601 (2019).

    Google Scholar 

  54. Rousk, J., Brookes, P. C. & Bååth, E. The microbial PLFA composition as affected by pH in an arable soil. Soil Biol. Biochem 42, 516–520 (2010).

    Google Scholar 

  55. Wang, C., Sun, Y., Chen, H. Y. H. & Ruan, H. Effects of elevated CO2 on the C:N stoichiometry of plants, soils, and microorganisms in terrestrial ecosystems. Catena 201, 105219 (2021).

    Google Scholar 

  56. Sun Y. et al. Coherent responses of terrestrial C:N stoichiometry to drought across plants, soil, and microorganisms in forests and grasslands. Agric. For. Meteorol. 292–293, 108104 (2020).

  57. Sun, Y., Wang, C., Chen, H. Y. H. & Ruan, H. Responses of C:N stoichiometry in plants, soil, and microorganisms to nitrogen addition. Plant Soil 456, 277–287 (2020).

    Google Scholar 

  58. Xu, X., Thornton, P. E. & Post, W. M. A global analysis of soil microbial biomass carbon, nitrogen and phosphorus in terrestrial ecosystems. Glob. Ecol. Biogeogr. 22, 737–749 (2013).

    Google Scholar 

  59. Zhao, M. Y., Liu, X. Y. & Zhang, X. Y. A review of research advances on carbon sinks in farmland ecosystem. Acta Ecol. Sin. 516, 2710–2710 (2022).

    Google Scholar 

  60. Lange, M. et al. Plant diversity increases soil microbial activity and soil carbon storage. Nat. Commun. 6, 6707 (2015).

    Google Scholar 

  61. Dijkstra, F. A. et al. Climate change alters stoichiometry of phosphorus and nitrogen in a semiarid grassland. N. Phytol. 196, 807–815 (2012).

    Google Scholar 

  62. Li, S. et al. Phosphorus limitation regulates the responses of microbial carbon metabolism to long-term combined additions of nitrogen and phosphorus in a cropland. Soil Biol. Biochem. 200, 109614 (2025).

    Google Scholar 

  63. Gu, X., Zhang, F., Xie, X., Cheng, Y. & Xu, X. Effects of N and P addition on nutrient and stoichiometry of rhizosphere and non-rhizosphere soils of alfalfa in alkaline soil. Sci. Rep. 13, 12119 (2023).

    Google Scholar 

  64. Cui, Y. et al. Extracellular enzyme stoichiometry reveals the carbon and phosphorus limitations of microbial metabolisms in the rhizosphere and bulk soils in alpine ecosystems. Plant Soil 458, 7–20 (2019).

    Google Scholar 

  65. Cleveland, C. C. & Liptzin, D. C. :N:P stoichiometry in soil: is there a “Redfield ratio” for the microbial biomass?. Biogeochemistry 85, 235–252 (2007).

    Google Scholar 

  66. Kuzyakov, Y. & Blagodatskaya, E. Microbial hotspots and hot moments in soil: concept and review. Soil Biol. Biochem. 83, 184–199 (2015).

    Google Scholar 

  67. Ning, Q. et al. Carbon limitation overrides acidification in mediating soil microbial activity to nitrogen enrichment in a temperate grassland. Glob. Change Biol. 27, 5976–5988 (2021).

    Google Scholar 

  68. Puissant, J. Does the C:N:P 1:1:1 ratio hold? Examining log-transformation bias in enzyme stoichiometry. Glob. Change Biol. 31, e70228 (2025).

    Google Scholar 

  69. Mori, T. The problem is not how we calculate enzyme stoichiometry threshold—It is that we calculate it. Glob. Change Biol. 31, e70519 (2025).

    Google Scholar 

  70. Zou, X. Calcium as a confounding variable in phosphorus attribution: a commentary on Hu et al. (2025). Soil Environ. Health 3, 100172 (2025).

    Google Scholar 

  71. Burda, B. U., O’Connor, E. A., Webber, E. M., Redmond, N. & Perdue, L. A. Estimating data from figures with a Web-based program: Considerations for a systematic review. Res. Synth. Methods 8, 258–262 (2017).

    Google Scholar 

  72. Hijmans, R. J., Cameron, S. E., Parra, J. L., Jones, P. G. & Jarvis, A. Very high resolution interpolated climate surfaces for global land areas. Int. J. Climatol. 25, 1965–1978 (2005).

    Google Scholar 

  73. Han, S. et al. Soil aggregate size-dependent relationships between microbial functional diversity and multifunctionality. Soil Biol. Biochem. 154, 10843 (2021).

    Google Scholar 

  74. Liu, M. et al. Unprotected carbon dominates decadal soil carbon increase. Nat. Commun. 16, 2008 (2025).

    Google Scholar 

  75. Bates, D., Mächler, M., Bolker, B. M. & Walker, S. C. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48 (2015).

    Google Scholar 

  76. R Development Core Team. R: a language and environment for statistical computing. R Foundation for Statistical Computing, (Vienna, 2024).

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Acknowledgements

This study was supported by the National Key Research and Development Program of China (No. 2023YFD2200404, 2021YFD2200403); the Key Project of the Open Competition in Jiangsu Forestry (LYKJ【2022】01); the Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX24_1358); the Natural Science Foundation of Jiangsu Province (BK20250706); and the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (24KJB220005).

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H.L., H.R., H.C., T.R., and X.Z. led the writing of the manuscript and designed the research; H.L. and T.R. collected the data; H.L, J.L., and H.C. analyzed the data. All authors contributed significantly to the drafts and gave final approval for publication.

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Tingting Ren or Honghua Ruan.

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Liu, H., Ren, T., Liao, J. et al. Nitrogen deposition alleviates phosphorus-induced imbalances in soil enzyme stoichiometry.
Commun Earth Environ (2025). https://doi.org/10.1038/s43247-025-03115-1

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