不同温度影响下除铀厌氧微生物群落结构特征解析曾涛涛

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2018,38(11)4261~4268ChinaEnvironmentalScience1*,1,1,1,1,1,1,2(1.,421001;2.,421001),152535.48h7.2mg/L,35(99.1%0.3%)1525.(43).,1525(Firmicutes),61.7%63.3%,35(Proteobacteria),68.0%.,1525Trichococcus;35,Klebsiella,52%,Proteiniclasticum(11%)Clostridium(8%)Acinetobacter(6%)Enterobacteriaceae(5%)Citrobacter(4)Sedimentibacter(4%)Desulfovibrio(3%).,.35.KlebsiellaTrichococcus,()().X172A10006923(2018)11426108Characterizationofmicrobialcommunitysuccessionforuraniumremovalunderdifferenttemperature.ZENGTao-tao1*,GAOXiang1,MOGuan-hai1,WANGGuo-hua1,LIUHai-yan1,XIEShui-bo1,2(1.HunanProvinceKeyLaboratoryofPollutionControlandResourcesReuseTechnology,UniversityofSouthChina,Hengyang421001,China;2.KeyDisciplineLaboratoryforNationalDefenceforBiotechnologyinUraniumMiningandHydrometallurgy,UniversityofsouthChina,Hengyang421001,China).ChinaEnvironmentalScience,2018,38(11)4261~4268AbstractInordertoinvestigatetheeffectofthetemperatureonmicrobialcommunityofuraniumremovalconsortium,themicrobialcompositionandabundanceoftheanaerobicbacteriainthesamplesunder15,25and35weredetectedusinghigh-throughputsequencingtechnique.Afterexposedininitial7.2mg/Luraniumfor48h,aremovalefficiencyof99.1%±0.3%wasobtainedforthesampleunder35Ԩ,whichismuchhigherthanthatin15and25Ԩ.Amajorityofcommonbacterialgenerawerefoundinthreesamplesunderdifferenttemperatures(43genera).Atthephylumlevel,Firmicutespredominatedinthe15℃and25℃samples,withtheproportionsof61.7%and63.3%,respectively.WhileProteobacteriawasthedominantphyluminthe35samplewhoseproportionwasashighas68.0%.Atthegenuslevel,thefirstmajorgenusinsamplesofboth15°Cand25°CwasTrichococcus.However,for35°Csample,itcontainedKlebsiellawiththehighestrelativeabundanceas52%,andotherdominantmicroorganismsincludingProteiniclasticum(11%),Clostridium(8%),Acinetobacter(6%),Enterobacteriaceae(5%),Citrobacter(4%),Sedimentibacter(4%),Desulfovibrio(3%).Therefore,thecompositionandabundanceofanaerobicconsortiumforuraniumremovalsignificantlyvariedwithtemperature.Apositivecorrelationbetweentemperatureandthemicrobialcommunityof35°CsamplewasobservedbyRDAanalysis.TheincreaseoftemperaturesignificantlypromotedtheabundanceofKlebsiella(positivecorrelation),butdramaticallyreducedthatofTrichococcus(negativecorrelation).Keywordsuraniummicrobialcommunityhigh-throughputsequencingtemperature,,.,,[1].,[2][3][4].,,.[5]BiologECO,,100mg/kg;200mg/kg,.Sutcliffe[6],1500mg/kg,20180419(51408293,11475080);(2013XQD10)*,,biowater@126.comDOI:10.19674/j.cnki.issn1000-6923.2018.0475426238.OntiverosValencia[7],U().Li[8]pH(3.6~4.0)(200mM)U(VI),U(),GeothrixDesulfovibrioFerribacteriumGeobacter,Acidovorax.,,,.,.,,Alpha,.11.1,.[9]:2g/LMgSO4,5g/LNa3C6H5O7·2H2O(),1g/LCaSO4,1g/LNH4Cl,0.5g/LK2HPO4,3.5g/LC3H5O3Na(),1g/LYeastextract(),2mL/L(NH4)2Fe(SO4)2·6H2O(,5%).1mol/LNaOHpH7.5,(GI54TW,,,)12120min(,0.45m).5g100mL,30200r/min6d,10%.,[10]..1.2GB676886[11],U3O8(),1g/LU().100mL7.2mg/LU(),0.1g,pH7.5.(QYC2112,,)1525353,5min,200r/min48h.8000g5min,0.45μm,5Brpadap[12],U().1.3DNAPCR,,(PBS,pH7.0)3,OMEGAsoilDNAKit(OMEGABioTek,USA)DNA.338F/806R16SrDNAV3+V4[13].PCR20μL,FastpfuDNA(TransGen,),PCR(ABIGeneAmp®9700,ABI,USA).953min;,9530s,5530s,7245s,27;7210min.3,2%PCR,AxyPrepDNA(AxygenBiosciences,Axygen,USA)PCR.1.4,PCRQuantiFluor™ST(Promega,USA).Miseq,IlluminaMiSeqPE(Illumina,USA).,,.1.5(30820),97%[14],(OTU),Usearch(vsesion7.1)OTU.R,Venn,[15].RDPclassifier97%OTU,Silva16SrRNA,(Phylum)(Genus),.Fisher,3515,Rstats.114263(RDA),RveganRDA.NCBISRA,PRJNA490608.22.1,1000,1a,.48h,15,30.6%2.4%;25,42.7%2.2%;35,99.1%0.3%..,.(a)020406080100U(VI)(%)()1525351(a)(b)Fig.1Morphology(a)anduraniumremovalperformance(b)ofanaerobicconsortium2.2OTU1,1525353,441~460bp,421~440bp,442~444bp,PCR.,OTU8093,OTU[16],35.1Table1Highthroughputsequencingresultsoftheanaerobicconsortium()(bp)(bp)OUT421~440441~4601530820444721423603800.999525356184421063424983870.99963535500443978825710930.9993OTU,,;OTU,RankAbundance,2a.,35,.,[17].3(coverage)0.999,,[18].,1(2b).0204060801001E-30.010.11101000600012000180002400030000360000.00.20.40.60.81.0(b)(%)(a)OUT3525152RankAbundance(a)(b)Fig.2RankAbundance(a)andCoveragecurves(b)ofbacteria2.3426438Venn,[15].3Venn,3.152549,253546,351544;1525358912,43.32.3VennFig.3Vennanalysisbasedonbacterial“genus”level2Table2Thecompositionofuniquegenerauponthethreedifferenttemperatureconditions152535StreptophytaAnaerovoraxSolibacteralesComamonadaceaeBacillaceaePropionicimonasKaistobacterGeobacterGZKB119SphingobiumABS6ClostridialesAnaerolinaceaeLactococcusPhycisphaeralesGemmatimonadetesC111DysgonomonasBacteroidetesMogibacteriaceaeenvOPS12PseudomonasClostridiaceaeRhodospirillaceaeDok59DechloromonasABY1S247Coxiellaceae,,;.Venn,3.2.42.4.11%,3.1525,,(Firmicutes),61.7%63.3%;(Proteobacteria)(Bacteroidetes).35,,68%;,28.9%2.2%.3(%)Table3Bacterialcompositionsandabundancesatphylumlevel(%)()1561.727.211.00.12563.328.97.70.13528.968.02.20.9Hu[1]U(),Firmicutes.Suriya[19]Cauvery,Proteobacteria(47.5%)Bacteroidetes(22.4%)Firmicutes(14.6%).,,.,152535,,(3).2.4.21%,,4.3,Trichococcus(),,40%32%0.Clostridium()PorphyromonadaceaeCitrobacter().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