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Advances in the aquatic sciences
RESEARCH ARTICLE

Relationships between nitrogen removal processes and functional microorganisms in the rhizosphere soil in a horizontal surface flow constructed wetland

Yinuo Zhu A B , Jing Li A B C , Zhangjie Cai A B , Wei Li A B , Yinru Lei A B , Manyin Zhang A B and Lijuan Cui A B C
+ Author Affiliations
- Author Affiliations

A Institute of Wetland Research, Chinese Academy of Forestry, Beijing Key Laboratory of Wetland Ecological Function and Restoration, Xianshan Road, Haidian District, Beijing, 100091, PR China.

B Beijing Hanshiqiao National Wetland Ecosystem Research Station, Muyan Road, Shunyi District, Beijing, 101399, PR China.

C Corresponding authors. Email: jingli_2015@caf.ac.cn; lkyclj@126.com

Marine and Freshwater Research 70(11) 1603-1610 https://doi.org/10.1071/MF19033
Submitted: 1 February 2019  Accepted: 30 April 2019   Published: 1 August 2019

Abstract

Plant species could significantly affect the nitrogen removal processes mediated by microorganisms in constructed wetlands. However, the links between nitrogen removal processes in the rhizosphere and the related functional microorganisms in a horizontal surface flow constructed wetland in winter remain poorly understood. In this study we collected 24 rhizosphere soils from Typha orientalis and Phragmites australis to evaluate potential nitrogen removal activities, namely the potential nitrification rate (PNR) and denitrification enzyme activity (DEA), and their relationship with functional genes (i.e. nitrate reductase, nirS, and ammonia mono-oxygenase, amoA, of ammonia-oxidising archaea, AOA, and ammonia-oxidising bacteria, AOB) in denitrifiers and nitrifiers in winter. DEA and PNR were significantly higher in the rhizosphere soil of T. orientalis than P. australis, which was due to the higher abundance of nitrifiers and denitrifiers in the rhizosphere of T. orientalis. AOB were the major predictor of PNR in rhizosphere soil of T. orientalis, whereas AOA were more important for P. australis. In addition, denitrifiers containing the nirS gene were found to be the main drivers of DEA, and AOA and AOB also contributed to the denitrification process in the rhizosphere soil of both plants. Furthermore, the abundance of nitrifiers was significantly affected by the C : N ratio, soil organic matter and moisture, whereas the abundance of denitrifiers was affected by soil moisture and pH.

Additional keywords: denitrifiers, nitrifiers.


References

Brix, H. (1997). Do macrophytes play a role in constructed treatment wetlands? Water Research 35, 11–17.
Do macrophytes play a role in constructed treatment wetlands?Crossref | GoogleScholarGoogle Scholar |

Brodrick, S. J., Cullen, P., and Maher, W. (1988). Denitrification in a natural wetland receiving secondary treated effluent. Water Research 22, 431–439.
Denitrification in a natural wetland receiving secondary treated effluent.Crossref | GoogleScholarGoogle Scholar |

Calheiros, C. S. C., Duque, A. F., Moura, A., Henriques, I. S., Correia, A., Rangel, A. O. S. S., and Castro, P. M. L. (2009). Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment. Bioresource Technology 100, 3228–3235.
Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment.Crossref | GoogleScholarGoogle Scholar |

Carballeira, T., Ruiz, I., and Soto, M. (2016). Effect of plants and surface loading rate on the treatment efficiency of shallow subsurface constructed wetlands. Ecological Engineering 90, 203–214.
Effect of plants and surface loading rate on the treatment efficiency of shallow subsurface constructed wetlands.Crossref | GoogleScholarGoogle Scholar |

Chen, Z. J., Tian, Y. H., Zhang, Y., Song, B. R., Li, H. C., and Chen, Z. H. (2016). Effects of root organic exudates on rhizosphere microbes and nutrient removal in the constructed wetlands. Ecological Engineering 92, 243–250.
Effects of root organic exudates on rhizosphere microbes and nutrient removal in the constructed wetlands.Crossref | GoogleScholarGoogle Scholar |

Cookson, W. R., Cornforth, I. S., and Rowarth, J. S. (2002). Winter soil temperature (2–15°C) effects on nitrogen transformations in clover green manure amended or unamended soils: a laboratory and field study. Soil Biology & Biochemistry 34, 1401–1415.
Winter soil temperature (2–15°C) effects on nitrogen transformations in clover green manure amended or unamended soils: a laboratory and field study.Crossref | GoogleScholarGoogle Scholar |

Faußer, A. C., Hoppert, M., Walther, P., and Kazda, M. (2012). Roots of the wetland plants Typha latifolia, and Phragmites australis, are inhabited by methanotrophic bacteria in biofilms. Flora 207, 775–782.
Roots of the wetland plants Typha latifolia, and Phragmites australis, are inhabited by methanotrophic bacteria in biofilms.Crossref | GoogleScholarGoogle Scholar |

Francis, C. A., Santoro, A. E., Oakley, B. B., Beman, J. M., and Roberts, K. J. (2005). Ubiquity and diversity of ammonia-oxidizing archaea in water columns and sediments of the ocean. Proceedings of the National Academy of Sciences of the United States of America 102, 14683–14688.
Ubiquity and diversity of ammonia-oxidizing archaea in water columns and sediments of the ocean.Crossref | GoogleScholarGoogle Scholar | 16186488PubMed |

Fraser, L. H., Carty, S. M., and Steer, D. (2004). A test of four plant species to reduce total nitrogen and total phosphorus from soil leachate in subsurface wetland microcosms. Bioresource Technology 94, 185–192.
A test of four plant species to reduce total nitrogen and total phosphorus from soil leachate in subsurface wetland microcosms.Crossref | GoogleScholarGoogle Scholar | 15158511PubMed |

Guo, G. X., Deng, H., Qiao, M., Yao, H. Y., and Zhu, Y. G. (2013). Effect of long-term wastewater irrigation on potential denitrification and denitrifying communities in soils at the watershed scale. Environmental Science & Technology 47, 3105–3113.
Effect of long-term wastewater irrigation on potential denitrification and denitrifying communities in soils at the watershed scale.Crossref | GoogleScholarGoogle Scholar |

He, J. Z., Shen, J. P., Zhang, L. M., Zhu, Y. G., Zheng, Y. M., Xu, M. G., and Di, H. J. (2007). Quantitative analyses of the abundance and composition of ammonia-oxidizing bacteria and ammonia oxidizing archaea of a Chinese upland red soil under long-term fertilization practices. Environmental Microbiology 9, 2364–2374.
Quantitative analyses of the abundance and composition of ammonia-oxidizing bacteria and ammonia oxidizing archaea of a Chinese upland red soil under long-term fertilization practices.Crossref | GoogleScholarGoogle Scholar | 17686032PubMed |

Herrmann, M., Saunders, A. M., and Schramm, A. (2009). Effect of lake trophic status and rooted macrophytes on community composition and abundance of ammonia-oxidizing prokaryotes in freshwater sediments. Applied and Environmental Microbiology 75, 3127–3136.
Effect of lake trophic status and rooted macrophytes on community composition and abundance of ammonia-oxidizing prokaryotes in freshwater sediments.Crossref | GoogleScholarGoogle Scholar | 19304820PubMed |

Hou, L., Zheng, Y., Liu, M., Gong, J., Zhang, X., Yin, G., and You, L. (2013). Anaerobic ammonium oxidation (anammox) bacterial diversity, abundance, and activity in marsh sediments of the Yangtze Estuary. Journal of Geophysical Research. Biogeosciences 118, 1237–1246.
Anaerobic ammonium oxidation (anammox) bacterial diversity, abundance, and activity in marsh sediments of the Yangtze Estuary.Crossref | GoogleScholarGoogle Scholar |

Hu, H. W., Chen, D., and He, J. Z. (2015). Microbial regulation of terrestrial nitrous oxide formation: understanding the biological pathways for prediction of emission rates. FEMS Microbiology Reviews 39, 729–749.
Microbial regulation of terrestrial nitrous oxide formation: understanding the biological pathways for prediction of emission rates.Crossref | GoogleScholarGoogle Scholar | 25934121PubMed |

Jung, J., Yeom, J., Kim, J., Han, J., Lim, H. S., Park, H., Hyun, S., and Park, W. (2011). Change in gene abundance in the nitrogen biogeochemical cycle with temperature and nitrogen addition in Antarctic soils. Research in Microbiology 162, 1018–1026.
Change in gene abundance in the nitrogen biogeochemical cycle with temperature and nitrogen addition in Antarctic soils.Crossref | GoogleScholarGoogle Scholar | 21839168PubMed |

Kowalchuk, G. A., and Stephen, J. R. (2001). Ammonia-oxidizing bacteria: a model for molecular microbial ecology. Annual Review of Microbiology 55, 485–529.
Ammonia-oxidizing bacteria: a model for molecular microbial ecology.Crossref | GoogleScholarGoogle Scholar | 11544365PubMed |

Lai, W. L., Zhang, Y., and Chen, Z. H. (2012). Radial oxygen loss, photosynthesis, and nutrient removal of 35 wetland plants. Ecological Engineering 39, 24–30.
Radial oxygen loss, photosynthesis, and nutrient removal of 35 wetland plants.Crossref | GoogleScholarGoogle Scholar |

Leininger, S., Urich, T., Schloter, M., Schwark, L., Qi, J., Nicol, G. W., Prosser, J. I., Schuste, S. C., and Schleper, C. (2006). Archaea predominate among ammonia-oxidizing prokaryotes in soils. Nature 442, 806–809.
Archaea predominate among ammonia-oxidizing prokaryotes in soils.Crossref | GoogleScholarGoogle Scholar | 16915287PubMed |

Leto, C., Tuttolomondo, T., Bella, S. L., Leone, R., and Licata, M. (2013). Effects of plant species in a horizontal subsurface flow constructed wetland – phytoremediation of treated urban wastewater with Cyperus alternifolius L. and Typha latifolia L. in the west of Sicily (Italy). Ecological Engineering 61, 282–291.
Effects of plant species in a horizontal subsurface flow constructed wetland – phytoremediation of treated urban wastewater with Cyperus alternifolius L. and Typha latifolia L. in the west of Sicily (Italy).Crossref | GoogleScholarGoogle Scholar |

Li, L., Yang, Y., Tam, N. F. Y., Yang, L., Mei, X. Q., and Yang, F. J. (2013). Growth characteristics of six wetland plants and their influences on domestic wastewater treatment efficiency. Ecological Engineering 60, 382–392.
Growth characteristics of six wetland plants and their influences on domestic wastewater treatment efficiency.Crossref | GoogleScholarGoogle Scholar |

Li, J., Zheng, Y. M., Liu, Y. R., Ma, Y. B., Hu, H. W., and He, J. Z. (2014). Initial copper stress strengthens the resistance of soil microorganisms to a subsequent copper stress. Microbial Ecology 67, 931–941.
Initial copper stress strengthens the resistance of soil microorganisms to a subsequent copper stress.Crossref | GoogleScholarGoogle Scholar | 24682341PubMed |

Massaccesi, L., Benucci, G. M. N., Gigliotti, G., Cocco, S., Corti, G., and Agnelli, A. (2015). Rhizosphere effect of three plant species of environment under periglacial conditions (Majella Massif, central Italy). Soil Biology & Biochemistry 89, 184–195.
Rhizosphere effect of three plant species of environment under periglacial conditions (Majella Massif, central Italy).Crossref | GoogleScholarGoogle Scholar |

Meng, P., Hu, W., Pei, H., Hou, Q., and Ji, Y. (2014). Effect of different plant species on nutrient removal and rhizospheric microorganisms distribution in horizontal-flow constructed wetlands. Environmental Technology 35, 808–816.
Effect of different plant species on nutrient removal and rhizospheric microorganisms distribution in horizontal-flow constructed wetlands.Crossref | GoogleScholarGoogle Scholar | 24645463PubMed |

Morales, S. E., Cosart, T., and Holben, W. E. (2010). Bacterial gene abundances as indicators of greenhouse gas emission in soils. The ISME Journal 4, 799–808.
Bacterial gene abundances as indicators of greenhouse gas emission in soils.Crossref | GoogleScholarGoogle Scholar | 20182521PubMed |

Paranychianakis, N. V., Tsiknia, M., and Kalogerakis, N. (2016). Pathways regulating the removal of nitrogen in planted and unplanted subsurface flow constructed wetlands. Water Research 102, 321–329.
Pathways regulating the removal of nitrogen in planted and unplanted subsurface flow constructed wetlands.Crossref | GoogleScholarGoogle Scholar | 27379728PubMed |

Prosser, J. I., and Nicol, G. W. (2008). Relative contributions of archaea and bacteria to aerobic ammonia oxidation in the environment. Environmental Microbiology 10, 2931–2941.
Relative contributions of archaea and bacteria to aerobic ammonia oxidation in the environment.Crossref | GoogleScholarGoogle Scholar | 18973620PubMed |

Purkhold, U., Pommerening-Röser, A., Juretschko, S., Schmid, M. C., Koops, H. P., and Wagner, M. (2000). Phylogeny of all recognized species of ammonia oxidizers based on comparative 16S rRNA and amoA sequence analysis: implications for molecular diversity surveys. Applied and Environmental Microbiology 66, 5368–5382.
Phylogeny of all recognized species of ammonia oxidizers based on comparative 16S rRNA and amoA sequence analysis: implications for molecular diversity surveys.Crossref | GoogleScholarGoogle Scholar | 11097916PubMed |

Rotthauwe, J. H., Witzel, K. P., and Liesack, W. (1997). The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. Applied and Environmental Microbiology 63, 4704–4712.
The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations.Crossref | GoogleScholarGoogle Scholar | 9406389PubMed |

Saggaï, M. M., Ainouche, A., Nelson, M., Cattin, F., and Amrani, A. E. (2017). Long-term investigation of constructed wetland wastewater treatment and reuse: selection of adapted plant species for metaremediation. Journal of Environmental Management 201, 120–128.
Long-term investigation of constructed wetland wastewater treatment and reuse: selection of adapted plant species for metaremediation.Crossref | GoogleScholarGoogle Scholar | 28654800PubMed |

Sartoris, J. J., Thullen, J. S., Barber, L. B., and Salas, D. E. (2000). Investigation of nitrogen transformations in a southern California constructed wastewater treatment wetland. Ecological Engineering 14, 49–65.
Investigation of nitrogen transformations in a southern California constructed wastewater treatment wetland.Crossref | GoogleScholarGoogle Scholar |

Schreiber, F., Wunderlin, P., Kai, M. U., and Wells, G. F. (2012). Nitric oxide and nitrous oxide turnover in natural and engineered microbial communities: biological pathways, chemical reactions, and novel technologies. Frontiers in Microbiology 3, 372.
Nitric oxide and nitrous oxide turnover in natural and engineered microbial communities: biological pathways, chemical reactions, and novel technologies.Crossref | GoogleScholarGoogle Scholar | 23109930PubMed |

Shaw, L. J., Nicol, G. W., Smith, Z., Fear, J., Prosser, J. I., and Baggs, E. M. (2006). Nitrosospira spp. can produce nitrous oxide via a nitrifier denitrification pathway. Environmental Microbiology 8, 214–222.
Nitrosospira spp. can produce nitrous oxide via a nitrifier denitrification pathway.Crossref | GoogleScholarGoogle Scholar | 16423010PubMed |

Sims, A., Gajaraj, S., and Hu, Z. (2012). Seasonal population changes of ammonia-oxidizing organisms and their relationship to water quality in a constructed wetland. Ecological Engineering 40, 100–107.
Seasonal population changes of ammonia-oxidizing organisms and their relationship to water quality in a constructed wetland.Crossref | GoogleScholarGoogle Scholar |

Smith, M. S., and Tiedje, J. M. (1979). Phases of denitrification following oxygen depletion in soil. Soil Biology & Biochemistry 11, 261–267.
Phases of denitrification following oxygen depletion in soil.Crossref | GoogleScholarGoogle Scholar |

Stottmeister, U., Wiessner, A., Kuschk, P., Kappelmeyer, U., Ka¨stner, M., Bederski, O., Mu¨ller, R. A., and Moormann, H. (2003). Effects of plants and microorganisms in constructed wetlands for wastewater treatment. Biotechnology Advances 22, 93–117.
Effects of plants and microorganisms in constructed wetlands for wastewater treatment.Crossref | GoogleScholarGoogle Scholar | 14623046PubMed |

Tan, E., Hsu, T. C., Huang, X., Lin, H. J., and Kao, S. J. (2017). Nitrogen transformations and removal efficiency enhancement of a constructed wetland in subtropical Taiwan. The Science of the Total Environment 601–602, 1378–1388.
Nitrogen transformations and removal efficiency enhancement of a constructed wetland in subtropical Taiwan.Crossref | GoogleScholarGoogle Scholar | 28605856PubMed |

Throbäck, I. N., Enwall, K., Jarvis, A., and Hallin, S. (2004). Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE. FEMS Microbiology Ecology 49, 401–417.
Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE.Crossref | GoogleScholarGoogle Scholar | 19712290PubMed |

Trias, R., Ruiz-Rueda, O., Garcia-Lledo, A., Vilar-Sanz, A., Lopez-Flores, R., Quintana, X. D., Hallin, S., and Baneras, L. (2012). Emergent macrophytes act selectively on ammonia-oxidizing bacteria and archaea. Applied and Environmental Microbiology 78, 6352–6356.
Emergent macrophytes act selectively on ammonia-oxidizing bacteria and archaea.Crossref | GoogleScholarGoogle Scholar | 22706066PubMed |

Werker, A. G., Dougherty, J. M., Mchenry, J. L., Werker, A. G., Dougherty, J. M., Mchenry, J. L., and Loon, W. A. V. (2002). Treatment variability for wetland wastewater treatment design in cold climates. Ecological Engineering 19, 1–11.
Treatment variability for wetland wastewater treatment design in cold climates.Crossref | GoogleScholarGoogle Scholar |

Wu, H., Wang, X., He, X., Zhang, S., Liang, R., and Shen, J. (2017). Effects of root exudates on denitrifier gene abundance, community structure and activity in a micro-polluted constructed wetland. The Science of the Total Environment 598, 697–703.
Effects of root exudates on denitrifier gene abundance, community structure and activity in a micro-polluted constructed wetland.Crossref | GoogleScholarGoogle Scholar | 28456121PubMed |

Xiao, R., Chen, B., Liu, Y., Wang, C., Gu, J. D., Feng, H., Du, G. Z., and Ma, X. J. (2014). Higher abundance of ammonia oxidizing archaea than ammonia oxidizing bacteria and their communities in Tibetan alpine meadow soils under long-term nitrogen fertilization. Geomicrobiology Journal 31, 597–604.
Higher abundance of ammonia oxidizing archaea than ammonia oxidizing bacteria and their communities in Tibetan alpine meadow soils under long-term nitrogen fertilization.Crossref | GoogleScholarGoogle Scholar |

Yoshinari, T., and Knowles, R. (1976). Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria. Biochemical and Biophysical Research Communications 69, 705–710.
Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria.Crossref | GoogleScholarGoogle Scholar | 817722PubMed |

Zheng, Y. C., Wang, X. C., Xiong, J. Q., Liu, Y. J., and Zhao, Y. Q. (2014). Hybrid constructed wetlands for highly polluted river water treatment and comparison of surface- and subsurface-flow cells. Journal of Environmental Sciences (China) 26, 749–756.
Hybrid constructed wetlands for highly polluted river water treatment and comparison of surface- and subsurface-flow cells.Crossref | GoogleScholarGoogle Scholar |