当前位置:首页 > 商业/管理/HR > 质量控制/管理 > 除磷工艺中含氧条件对聚磷菌种群结构影响研究傅以钢
,,,,(, 200092):DNA,PCR,DGGE().,.(Proteobacteria)(Acidobacterium)16SrDNAV3,NCBI(),.,,,,.:;;16SrDNA;;:X172 :A :0250-3301(2008)02-0474-08:2007-02-06;:2007-05-15:“”(NXCET-05-0387):(1971~),,,,E-mail:yigangcr@gmail.comInfluenceofCommunityStructureofPhosphorusRemovingBacteriaUnderOxygenContaininProcessesForPhosphorusRemovalFUYi-gang,DAIRui,LIUHong-bo,ZHAOJian-fu,XIASi-qing(StateKeyLaboratoryofPollutionControlandResourcesReuse,CollegeofEnvironmentalScienceandEngineering,TongjiUniversity,Shanghai200092,China)Abstract:Itwasstudiedforcommunitystructureofmicroorganismsinthephosphorusremovalprocessesunderthecirculatingsituation,andanalyzedformicroorganismsstructureandbehaviorcharacteristicsbythemolecularbiologytechniquewithdirectobtainingofDNAfromsamplesofactivatedsludge,andbynestedPCRandDGGE.Itwasalsodeterminedcommunitystructureofmicroorganisms.ItwasanalyzedstructuresofProteobacteriaandAcidobacteriumby16SrDNAV3areagenefragmentssequencesinactivatedsludge.BycomparinggenesequencesintheNationalCenterofBiologicalInformation(NCBI),weredeterminedthekindsofpartofmicroorganisms.Analyzingthelowofchangesofpreponderantbacteriainanaerobicaerobicandanaerobicanoxicconditionstakestoknow,thatunderthestablesituationofphosphorusremoving,thesystemofmicroorganismsstructurecankeptmostlyconstant.Minorityracesthathavechangedinamountorkindhassomethingtodowiththevariationofoxygenlevelinthesystem,butstructuretotallycanadapttheenvironmentalconditionsoftheprocesses,whileitplacedindynamicvarieties.Keywords:Proteobacteria;Acidobacterium;16SrDNA;micro-ecosystem;anaerobic ,[1].,、,“”“”.,:;、、,,[2].,,,.,(),,,,,[3].,()()[4].,(Proteobacteria)、(Actinobacteria)、(Acidobacterium).、.,,,,.29220082 ENVIRONMENTALSCIENCEVol.29,No.2Feb.,2008DOI:10.13227/j.hjkx.2008.02.0271 1.1 DNA1.1.1 DNAANAO,SBR,,、(1).SBR,9000rmin1min,-30℃,DNA.1.,DNA.3SDNA.1 Fig.1 Technologyforphosphorusremoval1 Table1 Samplingplacesandnumerationofsamples 12SBR1()SBR2()ⅠⅡⅢⅣ1.1.2 DNA DNA0.5%,..:0.5%(0.5g,0.5×TBE100mL);:150V;:20min;(EB):1μgL;Marker()DNAMarkerλ-HindⅢdigest.1.2 PCR1.2.1 PCRPCRDGGE,PCR(nestedPCR).,DNA1PCR;1PCR,16SrDNAV32PCR.2PCR1PCR,12PCR,.1PCR:40μL,10×PCR5μL,4×dNTP1μL,10mmolL1μL,TaqDNA1,(DNA)1μL.2PCR:80μL,10×PCR10μL,4×dNTP2μL,10mmolL2μL,TaqDNA2,(1PCR)2μL.,.45s,PCR,.PCR2.PCR,95℃10min,72℃12min.2 PCRTable2 PrimersandprogramsofamplificationPCR℃s℃s℃sP63F,R1378r309560536072120[5]F243,R1378r359560636072120[6]1PCR31f,R1378r309560536072120[6,7]α-F203a,R1378r359460576072120[8]β-EUB341F,UNIV907r30946053607260[9]2PCRgc-P338F,P518r35944560457290[8]4752:1.2.2 PCRPCR1%,..:1%(1g,0.5×TBE100mL);:150V;:20min;(EB):1μgL;Marker()DNAMarkerDL2000.1.3 DGGE2PCR80μLDGGE().8%,0.75mm.:50%~65%.150V,1×TAE,60℃,7.5h.(0.5mgL)1×TAE,.DNA,,30μL,PCR,、.1.4 16SrDNA16SrDNA,pUCm-T,pUCm-T(pUCm-TVector)APCR..TaqDNAPCRPCRDNA3′1A.pUCm-T,3′1T.,pUCm-T2PCRAT,,PCRpUCm-T,.1.4.1 PCRPCR(50μL),1μL20mmolLdATP2.5μLTaq,72℃10min,PCRTA.1.4.2 50μL,5μLLigationBuffer,8μLpUCm-T,32μLPCR,5μLT4DNALigase.16℃2h.1.4.3 :-70℃100μL,,;2μL,;30min;42℃60s;2min;400μLLB,37℃200~250rmin1h;4000rmin5min,400μL,;90mm,IPTGX-gal.;37℃1h,.1.4.4 -DNApUCm-T,DNALacZ,b-a,X-galIPTG,.LB,37℃,260rmin5~6h,pUCm-TPCR.1.5 DGGEPCR、16SrDNA,.BLASTNCBI,.2 2.1 .4,1∶1∶1,1.2,(AN),(AO),.,:1,1..,.,(O2),,,.,COD,,CN、P[10,11].COD200mgL,TN40~60mgL,TP5~10mgL,C476 29N、P.,(ANAO)12∶8(20Lh),(HRT)6h(HRT1.5h),(SRT)15d,25%(ANAO;ANAO),COD49mgL,TN14.2mgL,0.7mgL,(PMLSS)5.7%,23℃.、0~0.1、0.2~0.32.5~3.0mgL.,.2.2 DNAPCR4DNA,DNA0.5%.DNAPCR.16SrDNA(Actinobacteria)、(Proteobacteria)、(Acidobacterium)DNA1PCR,12PCR.PCR,(Actinobacteria).,,,..1PCR2~4.12PCR,220bp(),16SrDNA.M:Marker;1:Ⅰ;2:Ⅱ;3:Ⅲ;4:Ⅳ;b,2 α-(α-Proteobacteria)PCRFig.2 ResultofPCRamplificationforα-ProteobacteriaSmartView,α-Proteobacteria、β-ProteobacteriaAcidobacterium16SrDNAPCR:1100、6003 β-(β-Proteobacteria)PCRFig.3 ResultofPCRamplificationforβ-Proteobacteria4 (Acidobacterium)PCRFig.4 ResultofPCRamplificationforAcidobacterium1300bp.2.3 16SrDNA2PCR,DGGE5~8.5,.,β-,α-.1:Ⅰ;2:Ⅱ;3:Ⅲ;4:Ⅳ,5 DGGEFig.5 ResultofDGGEfingerprintforcommonmicroorganisms4772:6 α-(α-Proteobacteria)DGGEFig.6 ResultofDGGEfingerprintforα-Proteobacteria7 β-(β-Proteobacteria)DGGEFig.7 ResultofDGGEfingerprintforβ-Proteobacteria 5~8DGGE,,,.β-,,.α-,,.,,8 (Acidobacterium)DGGEFig.8 ResultofDGGEfingerprintforAcidobacterium.,SBR1()SBR2(),,,.,2.5~8,.,.5,1、3、78;615α-Proteobacteria,;721、22、2526β-Proteobacteria;827、28、30、31Acidobacterium,.,54、5、9、10;612α-Proteobacteria;717β-Proteobacteria,1819β-Proteobacteria,.,4、5、9、10、17.2、6、11、13、14、16、20、23、24、25、29、32,4,,478 29 3 16SrDNADGGETable3 Sequencesof16SrDNAfragmentsforpartofmicroorganismsbytheDGGEfingerprintNCBI11165371170-26247-153388800814.BLASTQ2Bdellovibriosp.ETB99%Bdellovibriosp.L99%Bdellovibriosp.RO99%Bdellovibriosp.MTA99%21165373341-20881-52380411439.BLASTQ2Flavobacteriumsp.Rud11100%31165303011-28556-198673443110.BLASTQ4UnculturedgammaProteobacterium98%EnvironmentalcloneCC-598%Thiothrixsp.CT398%UnculturedThiothrixsp.98%Thiothrixsp.NKBI-C98%71165371923-23354-124615745607.BLASTQ2Bdellovibriosp.Ec13100%Bdellovibriosp.CHI100%Unculturedd
本文标题:除磷工艺中含氧条件对聚磷菌种群结构影响研究傅以钢
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