title: Characterization and quantification of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in a nitrogen-removing reactor using T-RFLP and qPCR.
abstract: Using ammonia monooxygenase alpha-subunit (amoA) gene and 16S rRNA gene, the community structure and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in a nitrogen-removing reactor, which was operated for five phases, were characterized and quantified by cloning, terminal restriction fragment length polymorphism (T-RFLP), and quantitative polymerase chain reaction (qPCR). The results suggested that the dominant AOB in the reactor fell to the genus Nitrosomonas, while the dominant AOA belonged to Crenarchaeotal Group I.1a in phylum Crenarchaeota. Real-time PCR results demonstrated that the levels of AOB amoA varied from 2.9 x 10(3) to 2.3 x 10(5) copies per nanogram DNA, greatly (about 60 times) higher than those of AOA, which ranged from 1.7 x 10(2) to 3.8 x 10(3) copies per nanogram DNA. This indicated the possible leading role of AOB in the nitrification process in this study. T-RFLP results showed that the AOB community structure significantly shifted in different phases while AOA only showed one major peak for all the phases. The analyses also suggested that the AOB community was more sensitive than that of AOA to operational conditions, such as ammonia loading and dissolved oxygen.
authors.affiliation: Environmental Biotechnology Laboratory, Department of Civil Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong, SAR, China. jintao@hkucc.hku.hk
authors.forename: Tao
authors.initials: T
authors.lastname: Jin
authors.forename: Tong
authors.initials: T
authors.lastname: Zhang
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Yan
mesh_terms: D000641:Ammonia / Q000378:metabolism*; D001105:Archaea / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics; D001419:Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D019149:Bioreactors / Q000382:microbiology*; D019641:DNA, Archaeal / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism*; D010088:Oxidoreductases / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D019843:Archaeal Proteins; D001426:Bacterial Proteins; D019641:DNA, Archaeal; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D000641:Ammonia; D010088:Oxidoreductases; C045582:ammonia monooxygenase; D009584:Nitrogen
references.citation: FEMS Microbiol Lett. 1997 May 1;150(1):65-73
references.pmid: 9163908
references.citation: Environ Microbiol. 2008 Nov;10(11):3002-16
references.pmid: 18973621
references.citation: Nucleic Acids Res. 2004 Feb 25;32(4):1363-71
references.pmid: 14985472
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 28;103(48):18296-301
references.pmid: 17114289
references.citation: Extremophiles. 2008 Jul;12(4):605-15
references.pmid: 18465082
references.citation: FEMS Microbiol Rev. 2009 Sep;33(5):855-69
references.pmid: 19453522
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 12;105(6):2134-9
references.pmid: 18250313
references.citation: Appl Environ Microbiol. 1997 Dec;63(12):4704-12
references.pmid: 9406389
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 15;103(33):12317-22
references.pmid: 16894176
references.citation: Pol J Microbiol. 2008;57(3):241-8
references.pmid: 19004245
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14683-8
references.pmid: 16186488
references.citation: J Bacteriol. 1988 Feb;170(2):720-6
references.pmid: 2448289
references.citation: Water Res. 2002 Mar;36(6):1439-48
references.pmid: 11996334
references.citation: Environ Microbiol. 2007 Sep;9(9):2364-74
references.pmid: 17686032
references.citation: Arch Microbiol. 2002 Feb;177(2):139-49
references.pmid: 11807563
references.citation: Nature. 2006 Aug 17;442(7104):806-9
references.pmid: 16915287
references.citation: Environ Microbiol. 2005 Dec;7(12):1985-95
references.pmid: 16309395
references.citation: Environ Microbiol. 2007 Sep;9(9):2220-33
references.pmid: 17686020
references.citation: Appl Environ Microbiol. 2002 Jan;68(1):245-53
references.pmid: 11772633
references.citation: Appl Environ Microbiol. 2003 Jun;69(6):3213-22
references.pmid: 12788718
references.citation: Appl Environ Microbiol. 1996 Feb;62(2):340-6
references.pmid: 8593039
references.citation: Syst Appl Microbiol. 1991;14:346-51
references.pmid: 11540073
references.citation: Environ Microbiol. 2009 Sep;11(9):2310-28
references.pmid: 19515200
references.citation: Appl Environ Microbiol. 2006 Aug;72(8):5643-7
references.pmid: 16885322
references.citation: FEMS Microbiol Ecol. 2007 Jun;60(3):501-12
references.pmid: 17391333
references.citation: Appl Environ Microbiol. 2004 Feb;70(2):1008-16
references.pmid: 14766583
references.citation: Appl Microbiol Biotechnol. 2007 May;75(1):211-21
references.pmid: 17256118
references.citation: J Appl Microbiol. 2005;99(3):629-40
references.pmid: 16108805
references.citation: Appl Environ Microbiol. 1997 Aug;63(8):3233-41
references.pmid: 9251210
references.citation: Water Res. 2004 Aug-Sep;38(14-15):3275-86
references.pmid: 15276744
references.citation: Environ Microbiol. 2008 Apr;10(4):1068-79
references.pmid: 18266758
references.citation: Chemosphere. 2008 Apr;71(9):1683-92
references.pmid: 18313099
references.citation: Nature. 2005 Sep 22;437(7058):543-6
references.pmid: 16177789
references.citation: Water Res. 2007 Mar;41(5):1110-20
references.pmid: 17254627
references.citation: Appl Environ Microbiol. 2006 Dec;72(12):7767-77
references.pmid: 17012598
references.citation: Water Sci Technol. 2003;48(3):17-24
references.pmid: 14518850
references.citation: Environ Microbiol. 2007 Apr;9(4):1001-16
references.pmid: 17359272
references.citation: Environ Microbiol. 2003 Aug;5(8):691-705
references.pmid: 12871236
references.citation: Appl Environ Microbiol. 2000 Dec;66(12):5368-82
references.pmid: 11097916
references.citation: J Appl Microbiol. 2007 May;102(5):1401-17
references.pmid: 17448175
references.citation: Appl Environ Microbiol. 1998 Sep;64(9):3480-5
references.pmid: 9726900
references.citation: J Appl Microbiol. 2009 Sep;107(3):970-7
references.pmid: 19486399