Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Microbial abundance and community in subsurface flow constructed wetland microcosms: role of plant presence


In this research, the role of plants in improving microorganism growth conditions in subsurface flow constructed wetland (CW) microcosms was determined. In particular, microbial abundance and community were investigated during summer and winter in Phragmites australis-planted CW microcosms (PA) and unplanted CW microcosms (control, CT). Results revealed that the removal efficiencies of pollutants and microbial community structure varied in winter with variable microbial abundance. During summer, PA comprised more dominant phyla (e.g., Proteobacteria, Actinobacteria, and Bacteroidetes), whereas CT contained more Cyanobacteria and photosynthetic bacteria. During winter, the abundance of Proteobacteria was >40 % in PA but dramatically decreased in CT. Moreover, Cyanobacteria and photosynthetic bacterial dominance in CT decreased. In both seasons, bacteria were more abundant in root surfaces than in sand. Plant presence positively affected microbial abundance and community. The potential removal ability of CT, in which Cyanobacteria and photosynthetic bacteria were abundant during summer, was more significantly affected by temperature reduction than that of PA with plant presence.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Ahn C, Gillevet P, Sikaroodi M (2007) Molecular characterization of microbial communities in treatment microcosm wetlands as influenced by macrophytes and phosphorus loading. Ecol Indic 7:852–863

  2. Allen WC, Hook PB, Biederman JA, Stein OR (2002) Temperature and wetland plant species effects on wastewater treatment and root zone oxidation. J Environ Qual 31:1010–1016

  3. Ansola G, Arroyo P, Sáenz de Miera LE (2014) Characterisation of the soil bacterial community structure and composition of natural and constructed wetlands. Sci Total Environ 473:63–71

  4. APHA (2001) In: Frances, P.D., Keith, I. (Eds.), Compendium of Methods for the Microbiological Examination of Foods. Washington, DC

  5. Armstrong W (1978) Root aeration in the wetland condition. Plant life Anaerobic Environ 1:197

  6. Armstrong J, Armstrong W (1990) Light-enhanced convective throughflow increases oxygenation in rhizomes and rhizosphere of Phragmites australis (Cav.) Trin. ex Steud. New Phytol 114:121–128

  7. Baptista JC, Davenport RJ, Donnelly T, Curtis TP (2008) The microbial diversity of laboratory-scale wetlands appears to be randomly assembled. Water Res 42:3182–3190

  8. Barbosa MJ, Rocha J, Tramper J, Wijffels RH (2001) Acetate as a carbon source for hydrogen production by photosynthetic bacteria. J Biotechnol 85:25–33

  9. Bilgin M, Şimşek İ, Tulun Ş (2014) Treatment of domestic wastewater using a lab-scale activated sludge/vertical flow subsurface constructed wetlands by using < i > Cyperus alternifolius</i> Ecol Eng 70:362–365

  10. Braeckevelt M, Rokadia H, Imfeld G, Stelzer N, Paschke H, Kuschk P, Kästner M, Richnow H-H, Weber S (2007) Assessment of in situ biodegradation of monochlorobenzene in contaminated groundwater treated in a constructed wetland. Environ Pollut 148:428–437

  11. Brix H (1997) Do macrophytes play a role in constructed treatment wetlands? Water Sci Technol 35:11–17

  12. Brix H, Schierup H-H (1990) Soil oxygenation in constructed reed beds: the role of macrophyte and soil-atmosphere interface oxygen transport. Constructed wetlands in water pollution control, 53–66

  13. Calheiros C, Teixeira A, Pires C, Franco A, Duque A, Crispim L, Moura S, Castro P (2010) Bacterial community dynamics in horizontal flow constructed wetlands with different plants for high salinity industrial wastewater polishing. Water Res 44:5032–5038

  14. Collins B, McArthur JV, Sharitz RR (2004) Plant effects on microbial assemblages and remediation of acidic coal pile runoff in mesocosm treatment wetlands. Ecol Eng 23:107–115

  15. Dan TH, Quang LN, Chiem NH, Brix H (2011) Treatment of high-strength wastewater in tropical constructed wetlands planted with Sesbania sesban: horizontal subsurface flow versus vertical downflow. Ecol Eng 37:711–720

  16. Di HJ, Cameron KC, Shen JP, Winefield CS, O’Callaghan M, Bowatte S, He JZ (2010) Ammonia-oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions. FEMS Microbiol Ecol 72:386–394

  17. Dong X, Reddy GB (2010) Soil bacterial communities in constructed wetlands treated with swine wastewater using PCR-DGGE technique. Bioresour Technol 101:1175–1182

  18. Drew M, Jackson M, Giffard S (1979) Ethylene-promoted adventitious rooting and development of cortical air spaces (aerenchyma) in roots may be adaptive responses to flooding in Zea mays L. Planta 147:83–88

  19. Faulwetter JL, Burr MD, Parker AE, Stein OR, Camper AK (2013) Influence of season and plant species on the abundance and diversity of sulfate reducing bacteria and ammonia oxidizing bacteria in constructed wetland microcosms. Microb Ecol 65:111–127

  20. García J, Aguirre P, Barragán J, Mujeriego R, Matamoros V, Bayona JM (2005) Effect of key design parameters on the efficiency of horizontal subsurface flow constructed wetlands. Ecol Eng 25:405–418

  21. Garrity G, Bell J, Lilburn T (2006) Rhodobacteraceae fam. nov. List of New Names and New Combinations Previously Effectively, but not Validly, Published, 1-6

  22. Ge Y, Zhang C, Jiang Y, Yue C, Jiang Q, Min H, Fan H, Zeng Q, Chang J (2011) Soil microbial abundances and enzyme activities in different rhizospheres in an integrated vertical flow constructed wetland. CLEAN–Soil, Air, Water 39:206–211

  23. Gong H-L, Shi Y, Zhou L, Wu C-P, Cao P-Y, Tao L, Xu C, Hou D-S, Wang Y-Z (2013) The composition of microbiome in larynx and the throat biodiversity between laryngeal squamous cell carcinoma patients and control population. PLoS One 8:e66476

  24. Harper L, Langdale G, Giddens J (1987) Nitrogen cycling in a wheat crop: soil, plant, and aerial nitrogen transport. Agron J 79:965–973

  25. Hooper D, Chapin Iii F, Ewel J, Hector A, Inchausti P, Lavorel S, Lawton J, Lodge D, Loreau M, Naeem S (2005) Effects of biodiversity on ecosystem functioning: a consensus of current knowledge. Ecol Monogr 75:3–35

  26. Huang J, Cai W, Zhong Q, Wang S (2013) Influence of temperature on micro-environment, plant eco-physiology and nitrogen removal effect in subsurface flow constructed wetland. Ecol Eng 60:242–248

  27. Ike A, Toda N, Tsuji N, Hirata K, Miyamoto K (1997) Hydrogen photoproduction from CO2-fixing microalgal biomass: application of halotolerant photosynthetic bacteria. J Ferment Bioeng 84:606–609

  28. Jamieson TS, Stratton GW, Gordon R, Madani A (2003) The use of aeration to enhance ammonia nitrogen removal in constructed wetlands. Can Biosyst Eng 45:1.9–1.14

  29. Jensen SI, Kühl M, Priemé A (2007) Different bacterial communities associated with the roots and bulk sediment of the seagrass Zostera marina. FEMS Microbiol Ecol 62:108–117

  30. Kadlec RH, Knight RL, Vymazal J, Brix H, Cooper P, Haberl R (2000) Constructed wetlands for pollution control: processes, performance, design and operation. IWA scientific and technical report No.8. IWA specialist group on use of macrophytes in water pollution control. IWA Publishing; 155 pp

  31. Kersters K, de Vos P, Gillis M, Swings J, Vandamme P, Stackebrandt E (2006) Introduction to the Proteobacteria. In: Dwarkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E (eds) The prokaryotes, vol 5, 3rd edn. Springer, New York, pp 3–37

  32. Luederitz V, Eckert E, Lange-Weber M, Lange A, Gersberg RM (2001) Nutrient removal efficiency and resource economics of vertical flow and horizontal flow constructed wetlands. Ecol Eng 18:157–171

  33. Münch C, Kuschk P, Rske I (2005) Root stimulated nitrogen removal: only a local effect or important for water treatment? Water Sci Technol 51:185–192

  34. Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700

  35. Pell M, Nyberg F (1989) Infiltration of wastewater in a newly started pilot sand-filter system: I. Reduction of organic matter and phosphorus. J Environ Qual 18:457–462

  36. Perfler R, Laber J, Langergraber G, Haberl R (1999) Constructed wetlands for rehabilitation and reuse of surface waters in tropical and subtropical areas—first results from small-scale plots using vertical flow beds. Water Sci Technol 40:155–162

  37. Reinhardt M, Mller B, Gchter R, Wehrli B (2006) Nitrogen removal in a small constructed wetland: an isotope mass balance approach. Environ Sci Technol 40:3313–3319

  38. Ruiz-Rueda O, Hallin S, Bañeras L (2009) Structure and function of denitrifying and nitrifying bacterial communities in relation to the plant species in a constructed wetland. FEMS Microbiol Ecol 67:308–319

  39. Sasaki K, Watanabe M, Suda Y, Ishizuka A, Noparatnaraporn N (2005) Applications of photosynthetic bacteria for medical fields. J Bio Bioeng 100:481–488

  40. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541

  41. Shelef O, Gross A, Rachmilevitch S (2013) Role of plants in a constructed wetland: current and new perspectives. Water 5:405–419

  42. Stein OR, Hook PB (2005) Temperature, plants, and oxygen: how does season affect constructed wetland performance? J Environ Sci Heal 40:1331–1342

  43. Stottmeister U, Wießner A, Kuschk P, Kappelmeyer U, Kästner M, Bederski O, Müller R, Moormann H (2003) Effects of plants and microorganisms in constructed wetlands for wastewater treatment. Biotechnol Adv 22:93–117

  44. Subashchandrabose SR, Ramakrishnan B, Megharaj M, Venkateswarlu K, Naidu R (2011) Consortia of cyanobacteria/microalgae and bacteria: biotechnological potential. Biotechnol Adv 29:896–907

  45. Sundberg C, Stendahl JSK, Tonderski K, Lindgren P-E (2007) Overland flow systems for treatment of landfill leachates—potential nitrification and structure of the ammonia-oxidising bacterial community during a growing season. Soil Biol Biochem 39:127–138

  46. Tanaka N, Yutani K, Aye T, Jinadasa K (2007) Effect of broken dead culms of Phragmites australis on radial oxygen loss in relation to radiation and temperature. Hydrobiologia 583:165–172

  47. Taylor CR, Hook PB, Stein OR, Zabinski CA (2011) Seasonal effects of 19 plant species on COD removal in subsurface treatment wetland microcosms. Ecol Eng 37:703–710

  48. Truu M, Juhanson J, Truu J (2009) Microbial biomass, activity and community composition in constructed wetlands. Sci Total Environ 407:3958–3971

  49. Vymazal J (2011) Plants used in constructed wetlands with horizontal subsurface flow: a review. Hydrobiol 674:133–156

  50. Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267

  51. Wang Q, Xie H, Zhang J, Liang S, Ngo HH, Guo W, Liu C, Zhao CC, Li H (2014) Effect of plant harvesting on the performance of constructed wetlands during winter: radial oxygen loss and microbial characteristics. Environ Sci Pollut Res. doi:10.1007/s11356-014-3966-5

  52. Watanabe K, Kodama Y, Harayama S (2001) Design and evaluation of PCR primers to amplify bacterial 16S ribosomal DNA fragments used for community fingerprinting. J Microbiol Methods 44:253–262

  53. Weber KP, Legge RL (2011) Dynamics in the bacterial community level physiological profiles and hydrological characteristics of constructed wetland mesocosms during start-up. Ecol Eng 37:666–677

  54. Wen Y, Chen Y, Zheng N, Yang D, Zhou Q (2010) Effects of plant biomass on nitrate removal and transformation of carbon sources in subsurface-flow constructed wetlands. Bioresour Technol 101:7286–7292

  55. Wießner A, Kappelmeyer U, Kuschk P, Kästner M (2005) Influence of the redox condition dynamics on the removal efficiency of a laboratory-scale constructed wetland. Water Res 39:248–256

  56. Wu H, Zhang J, Li P, Zhang J, Xie H, Zhang B (2011a) Nutrient removal in constructed microcosm wetlands for treating polluted river water in northern China. Ecol Eng 37:560–568

  57. Wu S, Zhang D, Austin D, Dong R, Pang C (2011b) Evaluation of a lab-scale tidal flow constructed wetland performance: oxygen transfer capacity, organic matter and ammonium removal. Ecol Eng 37:1789–1795

  58. Wu S-q, Chang J-j, Dai Y, Wu Z-b, Liang W (2013) Treatment performance and microorganism community structure of integrated vertical-flow constructed wetland plots for domestic wastewater. Environ Sci Pollut Res 20:3789–3798

  59. Yang L, Chang H-T, Huang M-NL (2001) Nutrient removal in gravel-and soil-based wetland microcosms with and without vegetation. Ecol Eng 18:91–105

  60. Ye L, Shao M-F, Zhang T, Tong AHY, Lok S (2011) Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing. Water Res 45:4390–4398

  61. Zhang D, Gersberg RM, Keat TS (2009) Constructed wetlands in China. Ecol Eng 35:1367–1378

  62. Zhang T, Shao M-F, Ye L (2011) 454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants. ISME J 6:1137–1147

  63. Zhi W, Ji G (2012) Constructed wetlands, 1991-2011: a review of research development, current trends, and future directions. Sci Total Environ 441:19–27

  64. Zhong F, Wu J, Dai YR, Zhang ZH,Cheng SP, Zhang Q (2014) Bacterial community analysis by PCR-DGGE and 454-pyrosequencing of horizontal subsurface flow constructed wetlands with front aeration. Appl Microbiol Biotechnol, 1–14

Download references


We gratefully acknowledge financial support by the National Science Foundation of China (21007032 and 21307078), Shandong Provincial Natural Science Foundation, China (2009ZRB019Y9), and the Independent Innovation Foundation of Shandong University (2012JC029 and 2014JC023).

Ethics statement

The work described has not been published before. The work is not under consideration for publication anywhere else, and its publication has been approved by all coauthors.

Author information

Correspondence to Jian Zhang.

Additional information

Responsible editor: Robert Duran

Electronic supplementary material

Below is the link to the electronic supplementary material.


(DOCX 19 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wang, Q., Xie, H., Ngo, H.H. et al. Microbial abundance and community in subsurface flow constructed wetland microcosms: role of plant presence. Environ Sci Pollut Res 23, 4036–4045 (2016). https://doi.org/10.1007/s11356-015-4286-0

Download citation


  • Constructed wetland
  • Summer
  • Winter
  • Microbial abundance
  • Microbial community