城市污水处理系统中氨氧化细菌种群结构研究于健

29320093ActaScientiaeCircumstantiaeVo.l29,No.3Mar.,2009:(No.20521140076);(No.2006DFA91870)SupportedbytheKeyInternationalCooperativeProjectofNSFC(No.20521140076)andtheInternationalScienceandTechnologyCooperativeProjectofMOST(No.2006DFA91870):(1982),,Emai:[e-mail];*(),Emai:[e-mail]:YUJian(1982),female,Emai:[e-mail];*Correspondingauthor,Emai:[e-mail],,,.2009.[J].,29(3):521-526YuJ,YangM,QiR,etal.2009.Communitystructuresofammoniaoxidizingbacteriaindifferentmunicipalwastewatertreatmentsystems[J].ActaScientiaeCircumstantiae,29(3):521-526于健1,2,杨敏1,2,*,齐嵘2,周军3,张昱2,刘如铟21.北京林业大学环境科学与工程学院,北京1000832.中国科学院生态环境研究中心环境水质学国家重点实验室,北京1000853.北京城市排水集团有限责任公司,北京100044:20080505:20090116:PCRDGGE()710(AOB),AOB,,9AOB.Nitrosomonasoligotropha/aestuariilikecluster.:;;PCRDGGE;:02532468(2009)0352106:X171:ACommunitystructuresofammoniaoxidizingbacteriaindifferentmunicipalwastewatertreatmentsystemsYUJian1,2,YANGMin1,2,*,QIRong2,ZHOUJun3,ZHANGYu2,LIURuyin21.CollegeofEnvironmentalScienceandEngineering,BeijingForestUniversity,Beijing1000832.StateKeyLaboratoryofEnvironmentalAquaticChemistry,ResearchCenterforEcoEnvironmentalSciences,CAS,Beijing1000853.BeijingDrainageGroupCo.Ltd,Beijing100044Received5May2008;accepted16January2009Abstract:PCRDGGEwasappliedtoinvestigatethecommunitystructuresofAmmoniaoxidizingBacteria(AOB)intentreatmentsystemsofsevendifferentMunicipalWastewaterTreatmentPlants(MWTPs)inBeijing.TheresultsindicatedthatintheseMWTPs,exceptforonesystemwhichhadpoornitrificationperformance,allofthesystemshadAOB.ButthediversityofAOBwaslow,andtheAOBphenotypesweremainlygroupedintotheNitrosomonasoligotropha/aestuariilikecluster.Keywords:MWTP;activatedsludge;PCRDGGE;AOB1(Introduction),,,,(AOB).,,.,.,AOB.,AOB(Nitrosomonassp.).,(Nitrosospirasp.)(Nitrosococcussp.)(Limpiyakornetal.,2004;Limpiyakornetal.,2006).(Fluorescenceinsitu29hybridization,FISH)PCRDGGE,(Lydmarketal.,2007;Otawaetal.,2006)(Solidsretentiontime,SRT)(Hallinetal.,2005;Limpiyakornetal.,2005)(Limpiyakornetal.,2005)AOB.,,,,AOB.,PCRDGGE710AOB,.2(Materialsandmethods)2.1样品的采集710,1.20074,.110Table1Descriptionof10wastewatertreatmentsystemsin7MWTPs/(m3d-1)SRT/dCOD/(mgL-1)/(mgL-1)/(mgL-1)/(mgL-1)B53000AO:+4.2547.954.890.0%56.249.911.3%F44000A2O:++15.5588.735.294.0%66.916.275.8%J152000OD:14.9411.136.091.2%52.63.393.8%W15000SBR:32.0-40.4--0.5-X615000A2O:++16.7393.652.686.6%52.24.990.6%Q1228000iA2O:++7.5574.071.287.6%50.815.769.1%Q2229000A2O:++12.8574.050.491.2%50.86.088.3%G1400000iA2O:++33.0295.439.586.6%63.724.062.4%G2140000AO():(+)()-295.4--63.7--G3140000AO():(+)()10295.4--63.7--2.2PCRDGGE测序2.2.1PCRDGGEPCRDGGEDNA.16SrRNA,GCclampCTO189fABCTO189fCCTO654r2(Kowalchuketal.,1997),.:955min,35(9445s,5745s,7245s),7210min;PCR800ngL-1(BovineSerumAlbumin,BSA).GCclampGC341f534r(primer3primer2)(Muyzeretal.,1993),:954min,20(9430s,61.540s03,7240s),15(9430s,5540s,7240s),7210min.PCRBSA.PCRDGGE.DGGEDCodeTMUniversalMutationDetectionSystem(Bio-Rad,),10%,30%~50%(),1TAE,60,25V20min,130V7h.DGGEQuantityOne(,2005).2.2.2切胶测序,DNA,341f(GCclamp,primer1)(Muyzeretal.,1993)534rPCR,().NCBIBLAST,,MEGA3.1(Kumaretal.,2004).5223:3(Results)3.1图谱结果AOBDGGE1(F;G2G3.1.)2.,CTO189fABCTO654rCTO189fCCTO654rDGGE.31DGGE.,10DGGE,10,.1,AB1AB3,AB2,J()AB5.2,C3,C1,BFJWQC2C4C5C6C7.3,G1G3G2,WFJ,XBQ1Q23.31DGGE(#1~#10BFJWXQ1Q2G1G2G3)Fig.3DendrogramgeneratedbyUPGMAclusteranalysiscomparisonofDGGEpatternsfromFig.1(#1~#10showsamplesB,F,J,W,X,Q1,Q2,G1,G2andG3,respectively)3.2序列结果3.2.1最大相似性结果AB1~AB6C1~C7,,2.2,AB192%,95%.,AB3AB5C3C4C65AOB(Nitrosomonas),B,AOB.(Rhodocyclaceae).3.2.2系统发育树结果MEGA3.1,4.AB1~AB6,C1~523292DGGE(2)Table2SequenceanalysisofbandsexercisedfromDGGEgels(Genusesappearingmorethantwiceareindicatedinbold)AB1PerchloratereducingbacteriumPR(AY265866)92%B/F/J/W/X/Q1/Q2/G1/G2/G3AB2Dechloromonassp.RCB(AY032610)Ferribacteriumcda1(Y17060)98%B/F/X/Q1/Q2/G1/G2/G3AB3Nitrosomonassp.Nm59(AY123811)97%F/J/W/X/Q1/Q2/G1/G2/G3AB4Dechloromonassp.EMB269(DQ413167)100%F/W/Q1/Q2/G1/G2/G3AB5NitrosomonasoligotrophaAS1(EF016119)Nitrosomonassp.Nm59(AY123811)Nitrosomonassp.Is32(AJ621027)Nitrosomonassp.Is79A3(AJ621026)NitrosomonasoligotrophaNm45(AJ298736)NitrosomonasaestuariiNm36(AJ298734)95%F/JAB6PerchloratereducingbacteriumPMC(AY265878)Dechloromonassp.HZ(AF479766)Dechloromonassp.PC1(AY126452)Dechloromonassp.JM(AF323489)98%W/Q1/Q2/G1/G2/G3C1Dechloromonassp.RCB(AY032610)Ferribacteriumcda1(Y17060)98%B/F/X/Q1/Q2C2Dechloromonassp.RCB(AY032610)Ferribacteriumcda1(Y17060)97%BC3Nitrosomonassp.Is79A3(AJ621026)95%F/J/W/X/Q1/Q2/G1C4Nitrosomonassp.Nm59(AY123811)96%F/W/Q1/Q2C5Dechloromonassp.EMB269(DQ413167)100%F/J/X/Q1/Q2/G1C6Nitrosomonassp.D27(U56626)95%W/X/Q1/Q2/G1C7PerchloratereducingbacteriumPMC(AY265878)Dechloromonassp.HZ(AF479766)Dechloromonassp.PC1(AY126452)Dechloromonassp.JM(AF323489)98%W/Q1/Q2/G14DGGE16SrRNAFig.4Phylogenictreefor16SrRNAestablishedwithsequencesfromexcisedDGGEbands5243:C7.4,CTODGGE,AB3C4,AB4C5,AB2C1C2,AB6C7.Cluster:Cluster1NitrosomonasAOB,Cluster2DechloromonasRhodocyclaceae.Cluster1AB3C4C3AB5Nitrosom