不同污水除磷工艺中变形杆菌群落结构的分析刘倩

INDUSTRIALWATER＆WASTEWATERVol．42No．6Dec.，2011α-1，1，2（1.，300072；2.，430010）MBR，，，。［1］，［2-3］，：PCR－DGGE4α－。，α－，，。，，MBR，，MBR。MBR，79．6％77．7％，50％～60％。，／，80％。MBR，47．5％～84．1％，。：；PCR－DGGE；16SrDNA；；；：X703.1：A：%1009－2455（2011）06－0020－04Analysisofcommunitystructuresofα-proteobacteriaindifferentphosphorusremovingsystemsLIUQian1，YANGXi-long1，2（1.CollegeofEnvironmentalScienceandEngineering,TianjinUniversity,Tianjin300072,China;2.CentralandSouthernChinaMunicipalEngineeringDesignandResearchInstitute,Wuhan430010,China）Abstract：Thecommunitystructuresofα-proteobacteriainfourdifferentphosphorusremovingsystemsunderthenormalrunningconditionwereinvestigatedthroughPCR-DGGEtechnology.Theresearchindicatedthat,thedemandofα-proteobacteriaaboutwaterqualityandtechnicalconditionwasnothigh,almostallofthebacteriastrainswhichconstitutethemainbodyofthephosphorusremovingbacteriacommunityinthesystemweredominantspecies.However,onthewhole,therewereobviousdifferencesamongvarioustreatmentsystems,MBRsystemcontainedmoreabundantα-proteobacteriacommunitythantraditionalactivatedsludgesystem;andmaybeduetothepresenceofspecificbacteria,thediversityindexoftheformeronewasgreaterthanthatofthelaterone.Thecommunitystrcturesimilaritieswere79.6%and77.7%separatedinaerobicandanoxicstagesofMBRsystem;however,thesimilarityratewasonly50%-60%betweenthetwostages.Thecommunitystructuressimilarityratewasabove80%inaerobicandanoxic／anaerobicseparatedwastewatertreatmentsystems.BetweenthetraditionaltreatmentsystemandtheMBRsystem,thecommunitysimilaritycoefficientwasatabout47.5%-84.1%,andthecommunitysimilarityofaerobicstagewasbetter.Keywords：proteobacteria;PCR-DGGE;16SrDNA;phosphorusremovingbacteria;microbialcommunitystructure;membranebioreactor：（07JCZDJC02100）：2011-09-16；：2011-10-25·20·1Tab.1OperationconditionsandnumerationofsamplesofwastewatertreatmentplantsAA／O－－C1：，C2：，C3：B（），C4：CMBR，（PVDF），0.2μm，，8min，2min，M1：，M2：DMBRM3：，M4：2Tab.2OperationconditionsofsamplingplantsA150～2502015～40520～45152～51～2B200～3506040~501545～6020~303～51～2CMBR140～20010100101001050.9DMBR130～2505025～40530～5081.5～41ρ（CODCr）／（mg·L－1）ρ（）／（mg·L－1）ρ（TN）／（mg·L－1）ρ（TP）／（mg·L－1）3Tab.3Operationconditionsofsamplingstagesρ（DO）／（mg·L－1）ρ（MLSS）／（mg·L－1）SV／％CODCrTNTPC14.330∶158206880~9361~7688~9565C21.3332025C30.8240019C44.030∶135304765~8025~5065~7580~95M14.824∶177309885~9385~9490~9681~86M23.0661097M34.624∶175607265~8078~8585~9550~65M41.8463031／%［4］。PCR－DGGE4（MBR）α－，MBR，，。11．14，MBR2。4、1～3。8，41，3，。，DNA。1．2DNA1．2．1500mL，30min，，50mL50mL，6～10℃、9165×g10min；，30mL，10min；，30mLTE，10min；15mL，：α-·21·INDUSTRIALWATER＆WASTEWATERVol．42No．6Dec.，20114PCRTab.4PCRprimersandamplificationprocedure／／℃／s／℃／s／℃／s1PCRF203α，R1378r3094605760721202PCRR518，F357-GC20109494606065～55556060727260601α-DGGEFig.1DGGEprofileofα-proteobacteriaindifferentsamplingplacesC1C2C3C4M1M2M3M4ABJCDEFGHI5α-DGGETab.5ClassificationofDGGEprofileofα-proteobacteriaα-A、B、C、D、EF、IG、H，5mLDNA［5］。1．2．2DNADNA－／／DNA。BioerTechnologyBioFluxDNA。1．3DNA16SrDNAV3PCRα－PCR［6］，95℃10min，72℃12min。PCR［7］，94℃5min，72℃8min，200．5℃。4。1．4PCRDGGEC．B．S．SCIENTIFICDGGE－2001PCR，8％，30％～60％。，1×TAE，10％1×TAEPCR20μL，150V，60℃6．5h［5-6］。1．5Shannon－WienerQuantityOneDGGE，Shannon－Wiener（H）。H：H=-Si=1ΣPilogPi=-Si=1Σ（ni／N）log（ni／N）（1）：Pi＝ni／N，ni，N。22．1α－DGGEα－DGGE1，5。51，5A、B、C、DE，CDα－，，，。，、E，。MBR，B，C、DE。2，A。Fα－M1M2，，［8］。I，，α－，。GHα－MBR，，，MBR，，，MBR。·22·6α-ShannonTab.6Shannonindexesofα-proteobacteriacommunityC1C2C3C4M1M2M3M4H0.4560.5480.5140.5400.6050.6140.5470.506C1C2C3C4M1M2M3M4C1C2C3C4M1M2M3M410074.086.759.281.167.078.451.210086.872.668.160.384.154.410069.780.364.982.058.010047.560.059.149.810052.779.662.810060.777.710066.21007α-Tab.7Similarityofα-proteobacteriacommunityindifferentsamplingplaces%2．2α－α－Shannon（H）6。6，α－。Aα－。MBRM1、M2M3、M4，，。，MBR，。2．3α－DGGEQuantityone、，7。7，C3C1，C2C1C2。，α－，。α－MBRM1M3、M2M480％，MBR，MBRα－。C1、M1、M3，C4，C4／。MBR，47．5％～84．1％，，，。3（1）A、B、CD4α－，5，，。α－，，MBR。（2）DGGE，α－47．5％～86．8％，，。MBR，，α－。：［1］，，，．［J］．，2008，29（2）：474－481．［2］AtkinsonBW，MudalyDD，BuxF．ContributionofPseudomonasspptophosphorusuptakeintheanoxiczoneofananaerobic－anoxic－aerobiccontinuousactivatedsludgesystem［J］．WaterScienceandTechnology，2001，43（1）：139－146．［3］．16SrDNA［D］．：，2007．［4］，，，．［J］．，2006，37（6）：4－7．［5］，，，．MBR［J］．，2010，30（1）：69－70．［6］GomesNCM，HeuerH，SchonfeldJ，etal．Bacterialdiversityoftherhizosphereofmaize（Zeamays）grownintropicalsoilstudiedbytemperaturegradientgelelectrophoresis［J］．PlantandSoil，2001，232（1－2）：167－180．［7］MuyzerG，WaalEC，UitterlindenAG．Profilingofcomplexmicrobialpopulationsbydenaturinggradientgelelectrophoresisanalysisofpolymerasechainreaction－amplifiedgenescodingfor16SrRNA［J］．AppliedandEnvironmentalMicrobiology，1993，59（3）：695－700．［8］．MBR［D］．：，2010．：（1985-），，，，，（）022－87402126（）[e-mail]。，：α-·23·