低DO条件下不同培养期颗粒污泥的PCRDGGE分析左金龙

DOPCＲ－DGGE150076DOSBＲ/A/OPCＲ－DGGEDO1mg/L。0．5mm。PCＲ－DGGE、。DGGE8。10～20d20～50d140d、。DOPCＲ－DGGEX703C1000－4602201515－0077－05E201355PCＲ-DGGEAnalysisofGranularSludgeduringDifferentCulturePeriodswithLowDOZUOJin-longJIANGLi-mingWANGWeiLIUZhi-weiLIBiaoTANGHan-qingSchoolofFoodScienceandEngineeringHarbinUniversityofCommerceHarbin150076ChinaAbstractInordertoinvestigatethebiologicalcharacteristicsofgranularsludgeunderlowDOgranularsludgeduringdifferentcultureperiodswasinvestigatedbyPCＲ-DGGEinthesequencebatchre-actorSBＲoperatedinanoxic/aerationA/OprocesswithmeanDOvalueof1mg/Laeration．Theresultsshowedthemeandiameterofgranularsludgewas0．5mminmaturephases．PCＲ-DGGEa-nalysisindicatedthatthemicrobialspeciesingranularsludgeweremoreabundantandzoogloeawereva-rious．TheeightbandswerecomparedwiththeirhighlyhomologousspeciesbysequencesofpredominantDGGEbandsindifferentphasesofthesludgegranulation．BasedonphylogenetictreetheCaldilineaaerophilacouldpromotesludgegranulationanditwasdominantandplayedaskeletonroleinactivatedsludgeduringtheearlystart10to20d．PseudomonasAcidovoraxtemperancouldcompletenitrifica-tionandMicrobacteriumginsengisollicoulddegradeorganicmattersotheremovaleffectofnitrogenandorganicmattercouldbeensuredduringtheinitialperiodinthesystem．Thenumberoffilamentousbacte-riaCaldilineaaerophilasignificantlyreducedduringthemediumperiod20to50dandtheaerobicdenitrificationspeciesPseudomonaspsychrophilabecamethedominantspeciesachievingbetterre-·77·311520158CHINAWATEＲ＆WASTEWATEＲVol．31No．15Aug．2015movaleffectofnitrogenandphosphorus．Thaueraalwaysoccupiedadominantpositionandtheorganicmatternitrogenandphosphoruscouldbeco-degradedwithPleomorphomonasoryzaeduringthegranularsludgematureperiod140dorso．KeywordslowDOgranularsludgebiologicalcharacteristicsPCＲ-DGGEDO1mg/L1～3。PCＲ－DGGE4～7。DO、、SBＲ16SrDNAPCＲ－DGGEDODO。11。1Fig．1Schematicdiagramofexperimentalequipment1.1pH7．1～7．8、COD260～370mg/L、NH+4－N34～51mg/L、PO3－4－P2～4mg/L。。1.2SBＲ。600mm200mm9L2．5L。10cm。。。25±1℃。/A/O5、600。46h0．5h、0．5h、3h、1h、0．5h、0．5h。10～15dMLSS2500～3000mg/L。1.31.3.1SBＲ、、2～3。5min3000r/min－20℃。T1～T10SBＲ15、20、150、145、140、110、105、85、80、50d。1.3.2DNAMoBioDNAPowerSoilDNAIsola-tionKitDNA5μLDNA1%1×TAEλ/HindⅢmarkerDNA1%110Vλ/HindⅢmarker。AlphalmagerHP。1.3.3PCＲDNAPCＲ16SrDNABA101F5'-TGGCGGACGGGTGAGTAA-3'BA534Ｒ5'-ATTACCGCGGCTGCTGG-3'。BA534Ｒ5'40bpGC。PCＲ450μLGeneAmpPCＲsystem9700·87·3115．watergasheat．comPCＲ。PCＲ94℃5min3094℃1min、56℃1min、72℃1min0．1℃72℃10min16℃PCＲ－20℃。4DL2000marker1%PCＲ。1.3.4DGGEBio-ＲadDcodeTMDGGE。①40%～60%7mol/L40%100%。②PCＲ5LPCＲ5μL2×。③150V、60℃、7．5h。。④。1.3.5①PCＲ1．5mLtip30μL4℃2h。DNAPCＲ。TpMD19－TDH5αPCＲ。②DNAABABI3730pUC。。BlastGenBankClustalWMega4．0。22.180d。0．2～0．4mmMLSS3000mg/LSVI42．86mL/g。20d。0．5mm。SVI26～30mL/gMLSS5000mg/L。2.2DNADNA2λ/HindⅢT1～T10。2Marker10DNA23kbpDNAPCＲ。2DNAFig．2MapofcrudeDNAafteragarosegelelectrophoresis2.3PCＲPCＲ3DL2000DNAsizestand-ardT1～T10。DNA500bp534－101+40bpGCclamp。·97·．watergasheat．comDOPCＲ－DGGE31153PCＲFig．3MapofPCＲproductafteragarosegelelectrophoresis2.4DGGE1016SrDNAV3PCＲDGGE4。。DNA。8、9、14、16。8、。4PCＲDGGEFig．4DGGEgelofPCＲproducts2.5DGGEDGGE3、4、6、7、9、14、16、17NCBIBlast1。1DGGETab．1SequencesofpredominantDGGEbandsindifferentphasesofsludgegranulation%3D06-G1311190262Caldilineaaerophila415964E06-G1311190274Acidovoraxtemperans4361006H05-G1311270216Curvibacterdelicatus432987H08-G1311270219Ｒhizobiumlupini393999H09-G1311270220Pleomorphomonasoryzae3939914A03-G1311220527Microbacteriumginsengisolli4139816B02-G1311220528Pseudomonaspsychrophila42710017E07-G1311190266Thaueraaminoaromatica4389918。3Caldilineaaerophila。4Acidovoraxtemperans、。6Curvibacterdeli-catusPHA。7Ｒhizobiumlupini。9PleomorphomonasoryzaePAHs。14Microbacteri-umginsengisoli。16Pseudomonaspsychrophila。17Thaueraaminoaromati-ca。Thauerasp．。2.6DGGE·08·3115．watergasheat．com10～20d。20～50d。140d、。85。516SrDNAFig．5Phylogenetictreebasedon16SrDNAofDGGEprofilesindifferentphasesofsludgegranulation3①DO1mg/LSVI26～30mL/gMLSS5000mg/L0．5mm。②、、、、、、95%、。③10～20d。④20～50d140d、。1．SBＲJ．200420840－42．2．J．2009251511－14．3．SBＲJ．2010616132－138．4．J．2006262183－187．5．16SrDNADGGEJ．20062771424－1428．6．J．20132971－4．7．D．2013．1970－。[e-mail]2015－03－16·18·．watergasheat．comDOPCＲ－DGGE3115