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Article

Six-Year Biochar Experiment Reduces Soil N2O Emissions in Eucalyptus Plantations: Associations with Microbial N-Cycle Genes

by
Yunhuang Luo
1,2,
Yuyi Shen
3,*,
Hao Shi
2,
Qiumei Teng
2,
Guangping Xu
2,4,*,
Liangliang Huang
1,
Junzhi Chu
1,
Jialin Liao
2,
Denan Zhang
2,
Kechao Huang
2,
Yingjie Sun
2,
Zhiwen Tan
5 and
Yu Cao
1
1
College of Environmental Science and Engineering, Guilin University of Technology, Guilin 541000, China
2
Guangxi Key Laboratory of Plant Conservation and Restoration Ecology in Karst Terrain, Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Academy of Sciences, Guilin 541000, China
3
Guangxi Key Laboratory of Functional Phytochemicals and Sustainable Utilization, Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Academy of Sciences, Guilin 541000, China
4
Guangxi Guilin Urban Ecosystem Observation and Research Station, Guilin 541000, China
5
College of Life Sciences, Guangxi Normal University, Guilin 541000, China
*
Authors to whom correspondence should be addressed.
Microorganisms 2026, 14(7), 1519; https://doi.org/10.3390/microorganisms14071519
Submission received: 1 June 2026 / Revised: 1 July 2026 / Accepted: 8 July 2026 / Published: 12 July 2026
(This article belongs to the Special Issue Soil Microbial Carbon/Nitrogen/Phosphorus Cycling: 2nd Edition)
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Abstract

Nitrous oxide (N2O) is a major greenhouse gas, and terrestrial ecosystems are among the primary sources of its emissions. Biochar is recognized as an effective soil amendment for mitigating N2O emissions, but its long-term residual effects and microbial mechanisms in subtropical plantations remain unclear. Therefore, this study evaluated the residual effects of Eucalyptus-derived biochar on soil N2O emissions six years after a single application and explored associations with nitrogen cycle functional genes. A field experiment was conducted in a Eucalyptus plantation in northern Guangxi with biochar applied at six rates (0–6% w/w). Soil N2O fluxes were measured in the fifth and sixth years (2022–2023); soil chemical parameters, soil enzyme activities, and N2O-related microbial functional genes (amoA, nirK, nirS and nosZ) abundance were analyzed. Biochar application significantly reduced ammonium nitrogen content but enhanced nitrate nitrogen content. Urease, protease, and sucrase activities increased, while nitrate reductase, nitrite reductase, and hydroxylamine reductase activities decreased. Furthermore, quantitative analysis revealed substantial variations in functional gene abundances. The abundance of ammonia-oxidizing archaea (AOA-amoA) exhibited a unimodal response, whereas ammonia-oxidizing bacteria (AOB-amoA) showed a robust dose-dependent accumulation. Notably, annual N2O emissions were suppressed by up to 35.2%, driven by a 3.4-fold increase in nosZ gene abundance and a significant reduction in the (nirK + nirS)/nosZ ratio. This mitigation was attributed to enhanced N2O consumption by nosZ-harboring denitrifiers and reduced heterotrophic ammonia oxidation. Overall, these findings highlight the pivotal role of long-term organic amendments in steering nitrogen transformation pathways, providing a theoretical basis for sustainable soil management in subtropical plantations.
Keywords: biochar; nitrous oxide (N2O); functional gene; nitrification; denitrification; Eucalyptus plantation biochar; nitrous oxide (N2O); functional gene; nitrification; denitrification; Eucalyptus plantation
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MDPI and ACS Style

Luo, Y.; Shen, Y.; Shi, H.; Teng, Q.; Xu, G.; Huang, L.; Chu, J.; Liao, J.; Zhang, D.; Huang, K.; et al. Six-Year Biochar Experiment Reduces Soil N2O Emissions in Eucalyptus Plantations: Associations with Microbial N-Cycle Genes. Microorganisms 2026, 14, 1519. https://doi.org/10.3390/microorganisms14071519

AMA Style

Luo Y, Shen Y, Shi H, Teng Q, Xu G, Huang L, Chu J, Liao J, Zhang D, Huang K, et al. Six-Year Biochar Experiment Reduces Soil N2O Emissions in Eucalyptus Plantations: Associations with Microbial N-Cycle Genes. Microorganisms. 2026; 14(7):1519. https://doi.org/10.3390/microorganisms14071519

Chicago/Turabian Style

Luo, Yunhuang, Yuyi Shen, Hao Shi, Qiumei Teng, Guangping Xu, Liangliang Huang, Junzhi Chu, Jialin Liao, Denan Zhang, Kechao Huang, and et al. 2026. "Six-Year Biochar Experiment Reduces Soil N2O Emissions in Eucalyptus Plantations: Associations with Microbial N-Cycle Genes" Microorganisms 14, no. 7: 1519. https://doi.org/10.3390/microorganisms14071519

APA Style

Luo, Y., Shen, Y., Shi, H., Teng, Q., Xu, G., Huang, L., Chu, J., Liao, J., Zhang, D., Huang, K., Sun, Y., Tan, Z., & Cao, Y. (2026). Six-Year Biochar Experiment Reduces Soil N2O Emissions in Eucalyptus Plantations: Associations with Microbial N-Cycle Genes. Microorganisms, 14(7), 1519. https://doi.org/10.3390/microorganisms14071519

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