不同污泥龄膜生物反应器内微生物的群落结构特征田园

28320153ＲesearchofEnvironmentalSciencesVol．28No．3Mar．2015．J．2015283453-459．TIANYuanDUANLiangSONGYonghuietal．CharacteristicsofmicrobialcommunitystructureinmembranereactorsunderdifferentsolidretentiontimesJ．ＲesearchofEnvironmentalSc[e-mail]．*[e-mail]122*21231．1000832．1000123．100083MBＲSＲT10、20、4080d4MBＲPCＲ-DGGEMBＲDNA．SＲTMBＲSＲTMBＲNH4+-NCODCr90%．DGGEMBＲSＲTMBＲSＲTMBＲShannon-WienerSＲTSＲT80dMBＲShannon-Wiener．SＲTMBＲArcobactersp．、betaproteobacteriumThiothrixsp．SＲTMBＲMBＲ．PCＲ-DGGEX7031001-6929201503-0453-07ADOI10.13198j．issn．1001-6929．2015．03．17CharacteristicsofMicrobialCommunityStructureinMembraneＲeactorsunderDifferentSolidＲetentionTimesTIANYuan12DUANLiang2*SONGYonghui2YANGLiu1LIShan231．CollegeofGeoscienceandSurveyingEngineeringChinaUniversityofMiningandTechnologyBeijing100083China2．UrbanWaterEnvironmentalＲesearchBaseChineseＲesearchAcademyofEnvironmentalSciencesBeijing100012China3．CivilandEnvironmentalEngineeringSchoolUniversityofScienceandTechnologyBeijingBeijing100083ChinaAbstractInordertocharacterizethemicrobialcommunitystructureinmembranereactorsMBＲsunderdifferentsolidretentiontimesSＲTfourparallellab-scaleMBＲswereoperatedatSＲT102040and80d．DNAfingerprintingwasanalyzedbyPCＲ-DGGEandexcisedforbandsequencing．TheresultsshowedthattheSＲTdidnotinfluencetheperformanceofMBＲtotreatthewastewater．AllfourMBＲswereabletoremovemorethan90%ofNH4+-NandCODCr．TheresultsofDGGEindicatedthatthemicrobialcommunityinMBＲschangedobviouslyoverthewholeoperation．ThebacterialcommunitystructurechangedinthesamewayinMBＲsatdifferentSＲTs．TheapparentspeedofthesamemicroorganisminMBＲswasaffectedbySＲT．TheShannon-WienerdiversityindexandrichnessvalueallincreasedastheSＲTincreasedwhilethemicrobialdiversityindexatSＲT80dwasthehighestamongthefourMBＲs．SequencingresultsshowedthattheadvantageousspeciesinMBＲoperatedatdifferentSＲTsweredifferent．Arcobactersp．betaproteobacteriumandThiothrixsp．werethedominantspeciesinallreactorsandplayedimportantrolesinthereactor'soperation．KeywordsMBＲPCＲ-DGGEmicrobialcommunitystructureMBＲ．、、、、1-4．MBＲ5-6．28MBＲ、HＲT、ρDO、SＲT、7．SＲT8-9．SＲTMBＲ10-12SＲT13-14．SＲTMBＲMBＲ．PCＲ-DGGE、、、15-18．PCＲ-DGGEDNA、Shannon-Wiener、SＲTMBＲMBＲ、、．11.1MBＲ7L14cm15cm．PVDF0.95mm、2.45mm0.3μm0.02m2．ρDO5mgL．d4SＲT10、20、40、80d．MBＲ1．1MBＲFig.1Schematicdiagramofmembranebioreactor505.4mgL30mgL50mgL、NH4Cl133.75mgL、MgSO4·7H2O71mgL、KH2PO430.8mgL、CaCl2·2H2O19.3mgL、FeSO4·7H2O17.4mgL、ZnSO4·7H2O0.132mgL、MnCl2·4H2O0.126mgL、CuCl2·2H2O0.07mgL、KI0.0332mgL、CoCl2·6H2O0.03mgL、H3BO30.0248mgL、NaMoO4·2H2O0.016mgL．．HＲT17h、．4．7d450mL3000rmin1min－80℃DNA．1.2DＲ2800HACH、ρCODCrρNH4+-N．ρCODCrDＲB200HACH150℃2mL150℃2h3～150mgL．ρNH4+-N0.1mL20min0.4～50.0mgL．1.3MBＲDNAMPBioFastDNASPINKitforSoil．DNA．DNAOD260280260280nm1.71～2.00DNAPCＲ19．DNAPCＲ．DNA27F5'-AGAGTTTGATCCTGGCTCAG-3'1492Ｒ5'-TACGGYTACCTTGTTCGCTT-3'．1400bp25μL12.5μL4543GreenMix2×Promega1μmol27F1492ＲDNA10～100ngddH2OPromega25μL．95℃2min3095℃、30s53℃、30s72℃、2min72℃10min．2%．PCＲ16SrＲNAgeneV3357F-GC5'-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGG-CCTACGGGAGGCAGCAG-3'“GC”518Ｒ5'-ATTACCGCGGCTGCTGG-3'．240bp．50μL0.25μLTaqTaKaＲa5μL10×PCＲbuffer4μLdNTPs1μmol357F-GC518Ｒ200ngddH2O50μL．95℃2min3595℃、30s55℃、30s72℃、1min72℃10min．2%．DGGE8%40%～60%65V1×TAE60℃620min．DGGESYBＲGoldinvitrogenUSA40minGel-DocGelDocXＲ+BIO-ＲADUSA．QuantityOneDGGE．Shannon-WienerH'H'=－∑niNlnniN1niiN．ＲＲ=S－1lnN2S．EE=H'lnS3DGGE20μLddH2O4℃．22μL357F518Ｒ．PCＲ2%．GenBankhttp．ncbi．nlm．nih．govblastBLAST．22.1MBＲMBＲ1．MBＲ．SＲTMBＲNH4+-NCODCr90%MBＲSＲTMBＲ20．1SＲTMBＲTable1TreatmentresultsoftheMBＲsoperatedatdifferentSＲTsSＲTd10204080ρmgL33.0～40.033.0～40.033.0～40.033.0～40.0NH4+-NρmgL0.5～4.30.8～3.60.4～4.80.6～4.2%97.096.397.698.4ρmgL397.0～448.0397.0～448.0397.0～448.0397.0～448.0CODCrρmgL7.0～45.59.3～52.611.2～43.86.4～50.3%94.293.694.395.42.2DGGESＲTDGGE2．21dDGGE．．63d．SＲT1-7、2-5、3-44-5SＲT55428．1-1、1-2、2-1、2-2、2-7、3-1、3-6、4-14-7．SＲTMBＲ1-3、2-3、3-24-2SＲTSＲT80d4-2MBＲ63SＲT40d3-242SＲT20d2-321SＲT10d1-3DUAN14．4-6、4-7SＲT80dMBＲ．3MBＲDGGE1-1～4-7．2SＲTMBＲDGGEFig.2DGGEprofileofbacteriainMBＲsoperatedatdifferentSＲTs3SＲTMBＲFig.3SimilaritycomparisonofbacterialDGGEinMBＲsoperatedatdifferentSＲTs6543．1d．STＲ10、20、40、80d1d105d26.7%、46.8%、25.5%28.1%1dMBＲ．2.32SＲTShannon-WienerShannon-Wiener．15．．Shannon-Wiener．42d．Shannon-WienerSＲTSＲT80dMBＲ．．2SＲTMBＲTable2DiversityindexesofbacteriainMBＲsoperatedatdifferentSＲTsSＲTdd121426384105100.830.660.650.740.760.78Shannon-Wiener200.810.740.530.710.730.82400.790.730.620.800.820.85800.810.730.640.830.840.88101.541.311.301.341.361.42201.521.341.041.531.321.52401.461.331.271.501.551.56801.511.321.211.521.511.57100.280.230.230.260.270.34200.390.260.210.250.260.30400.290.240.240.320.360.37800.270.230.230.380.370.342.4DGGEGenBank．90%3．3STＲMBＲ16SrDNATable316SrDNAsequencingresultsofsomebandsinMBＲsoperatedatdifferentSＲTsGenBank1-2、2-2AY509431Unculturedbetaproteobacterium1-3、2-3、3-2、4-2KF448118UnculturedArcobactersp．cloneB1-461-4、2-7、3-3KF945142UnculturedThiothrixsp．cloneDZ-916S1-5、2-5KF981602UnculturedSphaerotilussp．cloneDO3-11