Effects and mechanisms of attached biofilms on aquatic nitrogen transformation and N2O emission

ZHANG Guan-jie; ZHOU Ying-ping; XIAO Lin

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China Environmental Science ›› 2026, Vol. 46 ›› Issue (3) : 1617-1624.

Environmental Ecology

Effects and mechanisms of attached biofilms on aquatic nitrogen transformation and N₂O emission

ZHANG Guan-jie, ZHOU Ying-ping, XIAO Lin

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Abstract

The effects and mechanisms of high ammonium (NH₄⁺) stress on nitrogen transformation and nitrous oxide (N₂O) emissions of attached biofilms on different substrate were investigated. Under high ammonium-nitrogen conditions, the nitrification potential of epiphytic biofilms associated with Vallisneria natans and Ceratophyllum demersum increased by 27.1-fold and 10.2-fold, respectively, whereas denitrification was suppressed, resulting in nitrogen accumulation. High NH₄⁺ levels promoted N₂O emissions associated with nitrification; however, N₂O production during nitrification accounted for only up to 1.28% of that from denitrification, and denitrification was the main pathway of N₂O emissions. Under high NH₄⁺ stress, the increased abundances of nitrifying bacteria and genes but decreased denitrifiers contributed to changes in nitrogen transformation activities. Concurrently, the N₂O emission potential (nirS/nosZ and nirK/nosZ) increased by 90% and 59%, which enhanced N₂O emissions. Notably, artificial plant biofilms exhibited the highest nitrification and denitrification activities, whereas N₂O emission from attached biofilms on Vallisneria natans was only 65.92%~82.76% of those from artificial plants and Ceratophyllum demersum L., suggesting dual regulation by functional microbial composition and microenvironment.

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ammonium stress / attached biofilms / N₂O emissions / microbial composition / functional gene

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ZHANG Guan-jie, ZHOU Ying-ping, XIAO Lin. Effects and mechanisms of attached biofilms on aquatic nitrogen transformation and N₂O emission[J]. China Environmental Science. 2026, 46(3): 1617-1624

References

[1] Chen H, Pan H, XIAO S, et al. Nitrous oxide dominates greenhouse gas emissions from hydropower's reservoirs in China from 2020 to 2060 [J]. Water Research, 2025,279:123420.
[2] Xiao Q, Xu X, Zhang M, et al. Coregulation of nitrous oxide emissions by nitrogen and temperature in China's third largest freshwater lake (Lake Taihu) [J]. Limnology and Oceanography, 2019, 64(3):1070-1086.
[3] Audet J, Levi E E, Jeppesen E, et al. Nutrient enrichment—but not warming—increases nitrous oxide emissions from shallow lake mesocosms [J]. Limnology and Oceanography, 2024,9999:1-12.
[4] 王大玲,杨雨虹,贺惠,等.氨氧化微生物对硝化潜势和N₂O生成的贡献 [J]. 中国环境科学, 2024,44(12):6828-6837. Wang D L, Yang Y H, He H, et al. Contribution of ammonia-oxidizing microorganisms to nitrification potential and N₂O production [J]. China Environmental Science, 2024,44(12):6828-6837.
[5] Kuypers M M M, Marchant H K, Kartal B. The microbial nitrogen- cycling network [J]. Nature Reviews Microbiology, 2018,16:263.
[6] Huang Y, Deng M, LI L, et al. Freshwater salinization mitigated N₂O emissions in submerged plant-covered systems: Insights from attached biofilms [J]. Environmental Science & Technology, 2025,59(6):3205-3217.
[7] 梁淑雅,崔陈陈,胡思文,等.苦草附生细菌的分离鉴定及高效功能菌株筛选 [J]. 中国环境科学, 2024,44(5):2630-2641. Liang S Y, Cui C C, Hu S W, et al. Isolation and identification of epiphytic bacteria from Vallisneria natans and screening of functional strains [J]. China Environmental Science, 2024,44(5):2630-2641.
[8] Mu X, Lv X, Liu W, et al. Biofilms attached to Myriophyllum spicatum play a dominant role in nitrogen removal in constructed wetland mesocosms with submersed macrophytes: Evidence from 15N tracking, nitrogen budgets and metagenomics analyses [J]. Environmental Pollution, 2020,266:115203.
[9] Chen C, Yin G, Li Q, et al. Effects of microplastics on denitrification and associated N₂O emission in estuarine and coastal sediments: insights from interactions between sulfate reducers and denitrifiers [J]. Water Research, 2023,245:120590.
[10] Guo W, Liu T, Fan Z, et al. Aquatic plants dominate spatiotemporal dynamics of N₂O fluxes in small urban lake by regulating nutrient distribution and emission path [J]. Environmental Research, 2025, 274:121290.
[11] Yuan G, Fu H, Zhong J, et al. Nitrogen/carbon metabolism in response to NH₄⁺ pulse for two submersed macrophytes [J]. Aquatic Botany, 2015,121:76-82.
[12] Yuan G, Tan X, Guo P, et al. Linking trait network to growth performance of submerged macrophytes in response to ammonium pulse [J]. Water Research, 2023,229:119403.
[13] Zhang W, Li Q, Yang Y, et al. Joint toxicity mechanisms of perfluorooctanoic acid and sulfadiazine on submerged macrophytes and periphytic biofilms [J]. Journal of Hazard Materials, 2023,458:131910.
[14] Davidson T A, Audet J, Svenning J C, et al. Eutrophication effects on greenhouse gas fluxes from shallow-lake mesocosms override those of climate warming [J]. Global Change Biology, 2015,21(12):4449-4463.
[15] Yang L, Zhang S, Lv X, et al. Vallisneria natans decreased CH₄fluxes in wetlands: Interactions among plant physiological status, nutrients and epiphytic bacterial community [J]. Environmental Research, 2023, 224:115547.
[16] Guo S, Zhang S, Wang S, et al. Potamogeton crispus restoration increased the epiphytic microbial diversity and improved water quality in a micro-polluted urban river [J]. Environmental Pollution, 2023, 326:121485.
[17] Penning W E, Mjelde M, Dudley B, et al. Classifying aquatic macrophytes as indicators of eutrophication in European lakes [J]. Aquatic Ecology, 2008,42(2):237-251.
[18] Zhu Z, Yuan H, Wei Y, et al. Effects of ammonia nitrogen and sediment nutrient on growth of the submerged plant Vallisneria natans [J]. Clean Soil, Air, Water, 2015,43(12):1653-1659.
[19] Dou Y, Wang B, Chen L, et al. Alleviating versus stimulating effects of bicarbonate on the growth of vallisneria natans under ammonia stress [J]. International Journal of Environmental Science and Pollution Research, 2013,20(8):5281-5288.
[20] Qiu Z, Zhang S, DING Y, et al. Comparison of Myriophyllum Spicatum and artificial plants on nutrients removal and microbial community in constructed wetlands receiving WWTPs effluents [J]. Bioresource Technology, 2021,321:124469.
[21] Huang Y, Deng M, Zhou S, et al. Microbial mechanisms underlying the reduction of N₂O emissions from submerged plant covered system [J]. Water Research X, 2025,28:100314.
[22] Jeffrey S W, Humphrey G F. New spectrophotometric equations for determining chlorophylls a, b, c1and c2in higher plants, algae and natural phytoplankton [J]. Biochemie und Physiologie der Pflanzen, 1975,167(2):191-194.
[23] 李朝英,郑路,卢立华,等.测定植物全氮的H₂SO₄-H₂O₂消煮法改进 [J]. 中国农学通报, 2014,30(6):159-162. Li Z Y, Zheng L, Lu L H, et al. Improvement in the H₂SO₄-H₂O₂ digestion method for determining plant total nitrogen [J]. Chinese Agricultural Science Bulletin, 2014,30(6):159-162.
[24] 国家环境保护总局.水和废水监测分析方法 [M]. 北京:中国环境科学出版社, 2002. State Environmental Protection Administration. Standard methods for the examination of water and wastewater [M]. Beijing: China Environmental Science Press. 2002.
[25] Taylor A E, Zeglin L H, Wanzek T A, et al. Dynamics of ammonia- oxidizing archaea and bacteria populations and contributions to soil nitrification potentials [J]. The ISME Journal, 2012,6(11):2024-2032.
[26] Yang X, He Q, Guo F, et al. Nanoplastics disturb nitrogen removal in constructed wetlands: Responses of microbes and macrophytes [J]. Environmental Science & Technology, 2020,54(21):14007-14016.
[27] Yu L, Liu S, Jiang L, et al. Insight into the nitrogen accumulation in urban center river from functional genes and bacterial community [J]. PLoS One, 2020,15(9):e0238531.
[28] Cao T E, Ni L, Xie P, et al. Effects of moderate ammonium enrichment on three submersed macrophytes under contrasting light availability [J]. Freshwater Biology, 2011,56(8):1620-1629.
[29] Yan L, Mu X, Han B, et al. Ammonium loading disturbed the microbial food webs in biofilms attached to submersed macrophyte Vallisneria natans [J]. Science of the Total Environment, 2019,659: 691-698.
[30] Fu H, Yuan G X, Cao T, et al. Relationships between relative growth rate and its components across 11submersed macrophytes [J]. Journal of Freshwater Ecology, 2012,27(4):471-480.
[31] Gao J, Ren P, Zhou Q, et al. Comparative studies of the response of sensitive and tolerant submerged macrophytes to high ammonium concentration stress [J]. Aquatic Toxicology, 2019,211:57-65.
[32] Zhao Y, Liu H, Wang R, et al. Interactions between dicyandiamide and periphytic biofilms in paddy soils and subsequent effects on nitrogen cycling [J]. Science of The Total Environment, 2020,718:137417.
[33] Li X, Gao D, Li Y, et al. Increased nitrogen loading facilitates nitrous oxide production through fungal and chemodenitrification in estuarine and coastal sediments [J]. Environmental Science & Technology, 2023, 57(6):2660-2671.
[34] León-palmero E, Morales-baquero R, Reche I. P inputs determine denitrifier abundance explaining dissolved nitrous oxide in reservoirs [J]. Limnology and Oceanography, 2023,68(8):1734-1749.
[35] Sun X, Buchanan P J, Zhang I H, et al. Ecological dynamics explain modular denitrification in the ocean [J]. Proceedings of the National Academy of Sciences, 2024,121(52): e2417421121.
[36] Jiang C, Zhang S, Wang J, et al. Nitrous oxide (N₂O) emissions decrease significantly under stronger light irradiance in riverine water columns with suspended particles [J]. Environmental Science & Technology, 2023,57(48):19749-19759.
[37] Caranto J D, Lancaster K M. Nitric oxide is an obligate bacterial nitrification intermediate produced by hydroxylamine oxidoreductase [J]. Proceedings of the National Academy of Sciences, 2017,114(31): 8217-8222.
[38] Ju X, Sun H, Ruan C, et al. Prophage induction and quorum sensing enhance biofilm stability and resistance under ammonia-oxidizing bacteria-mediated oxidative stress [J]. Water Research, 2025,284: 124010.
[39] Venterea R T. Nitrite-driven nitrous oxide production under aerobic soil conditions: kinetics and biochemical controls [J]. Global Change Biology, 2007,13(8):1798-809.
[40] Liang X, Wang B, Gao D, et al. Nitrification regulates the spatiotemporal variability of N₂O emissions in a eutrophic lake [J]. Environmental Science & Technology, 2022,56(23):17430-17442.
[41] Liu Y, Gong L, Mu X, et al. Characterization and co-occurrence of microbial community in epiphytic biofilms and surface sediments of wetlands with submersed macrophytes [J]. Science of the Total Environment, 2020,715:136950.
[42] Wang Z, Liu Z, Wang J, et al. Characterizing algal-bacterial symbiotic biofilms: Insights into coexistence of algae and anaerobic microorganisms [J]. Bioresource Technology, 2024,406:130966.
[43] Wan H, Wang K, Luo X, et al. Algal-mediated nitrogen removal and sustainability of algal-derived dissolved organic matter supporting denitrification [J]. Bioresource Technology, 2024,407:131083.
[44] Yue X, Liu H, Wei H, et al. Reactive and microbial inhibitory mechanisms depicting the panoramic view of pH stress effect on common biological nitrification [J]. Water Research, 2023,231:119660.
[45] Dalsgaard T, Stewart F J, Thamdrup B, et al. Oxygen at nanomolar levels reversibly suppresses process rates and gene expression in anammox and denitrification in the oxygen minimum zone off northern chile [J]. mBio., 2014,5(6):e01966.