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cultivable microbial community in 2-km-deep, 20-million-year-old subseafloor coalbeds through ~1000 days anaerobic bioreactor cultivation hiroyuki imachi外文参考.pdf

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cultivable microbial community in 2-km-deep, 20-million-year-old subseafloor coalbeds through ~1000 days anaerobic bioreactor cultivation hiroyuki imachi外文参考.pdf

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cultivable microbial community in 2-km-deep, 20-million-year-old subseafloor coalbeds through ~1000 days anaerobic bioreactor cultivation hiroyuki imachi外文参考.pdf

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文档介绍:该【cultivable microbial community in 2-km-deep, 20-million-year-old subseafloor coalbeds through ~1000 days anaerobic bioreactor cultivation hiroyuki imachi外文参考 】是由【宝钗文档】上传分享,文档一共【16】页,该文档可以免费在线阅读,需要了解更多关于【cultivable microbial community in 2-km-deep, 20-million-year-old subseafloor coalbeds through ~1000 days anaerobic bioreactor cultivation hiroyuki imachi外文参考 】的内容,可以使用淘豆网的站内搜索功能,选择自己适合的文档,以下文字是截取该文章内的部分文字,如需要获得完整电子版,请下载此文档到您的设备,方便您编辑和打印。:..entificreportsopeNmunityin2-km-deep,20-million-year-oldsubseafloorcoalbedsthroughReceived:15October2018Accepted:9January2019~1000daysanaerobicbioreactorPublished:xxxxxxxxcultivationHiroyukiImachi1,2,EijiTasumi1,YoshihiroTakaki1,3,TatsuhikoHoshino2,4,FlorenceSchubotz5,ShuchaiGan5,Tzu-Hsuantu1,6,Yumisaito1,YukoYamanaka1,AkiraIjiri2,3,YoheiMatsui2,3,MasayukiMiyazaki1,YukiMorono2,4,Kentakai1,2,Kai-UweHinrichs5&FumioInagaki2,4,7munitiespersistinginsedimentsdownto~,,wepresentacultivationexperimentof2-km-munitiesin20-million-year-oldlignitecoalbedsusingacontinuous-flowbioreactoroperatingat40°-excitationmatricesspectroscopyandstableisotopeanalysestracedthetransformationofcoalbed-,theproductionofacetateand13C-ogetherwiththeincreaseandtransformationofhighmolecularweighthumicspointtoanactivelignite-(.,phylaProteobacteria,Firmicutes,Chloroflexi,Actinobacteria,Bacteroidetes,Spirochaetes,Tenericutes,Ignavibacteriae,andSBR1093).Theseresultsindicatethatactivationandadaptivegrowthof2-km-deepplishedusingacontinuous-flowbioreactor,,scientificoceandrillinghasdemonstratedthatnumerousmicrobesexistintheglobaldeepsubseafloorsediment,prisingapproximately1,-fueledmicrobialenergyrespiratoryactivityisextraordinarylow,×10?×10?14moles/e?/cell/yearbetweentheanoxiceasternequatorialPacificandtheoxicSouthPacificGyresediments,respectively3–-independentmolecularecologicalstudies(.,PCR-mediated16SrRNAandfunctionalgeneanalysis,ormetagenomics)oftheabove-mentionedsubseafloorsettingsshowedthattheyharbordiverse1DepartmentofSubsurfaceGeobiologicalAnalysisandResearch(D-SUGAR),JapanAgencyforMarine-EarthScienceandTechnology(JAMSTEC),Yokosuka,Kanagawa,237-0061,,JAMSTEC,Yokosuka,Kanagawa,237-0061,-generationResearchProtocolforSubmarineResources,JAMSTEC,Yokosuka,Kanagawa,237-0061,,JAMSTEC,Nankoku,Kochi,783-8502,,UniversityofBremen,D-28359,Bremen,,NationalTaiwanUniversity,Taipei,106,,JAMSTEC,Yokohama,Kanagawa,236-0001,.(email:******@)ScientificRepoRts|(2019)9:2305|https:///-019-38754-w1:..entificreports/munities,icallydistinctfromthoselivingintheEarth’ssurfaceenvi-ronments6–9;hence,theirphysiologyandmetabolicfunctionsstillremainlargelyunknown10,,-mentcoresamples,however,munitiestoconventionalbatch-,onlyasmallfractionofindigenousdeepmicrobescouldbeiso-latedthusfarfrom≥10mbelowseafloor(mbsf)sedimentsamples,whosemembersareprimarilyaffiliatedwiththephylaProteobacteria,Firmicutes,Actinobacteria,andBacteroidetesorEuryarchaeotageneraMethanoculleus,us,andMethanosarcina8,12–,binedwithnanometer-scalesecondaryionmassspectrometry(NanoSIMS)analysisconfirmedthatmorethan70%ofthetotalmicrobialcellsareviable,despitehavingveryslowbiomassturnoverrates15,,cultivationofdeepsubseafloormicrobesthroughbatch-typetechniquesmaybeimpededbytheirextraordinarilylowmeta-bolicactivityunderenergy-limitedconditions5and/orthe“substrate-accelerateddeath”phenomenon,,-pressureanaerobicenrichmentsystem(.,DeepIsoBUG)forgashydrate-essfullyobtainedsomeanaerobicbacteria(.,generaAcetobacteriumandClostridium).Imachiandco-workers(2011,2014,2017)19–21appliedacontinuous-flowbioreactorcultiva-ethelimitationofbatch-essfullyenrichedpreviouslyuncul--flowhangingsponge(DHS)reactorsystem,,--flowreactorstherebyanismsinacontrolledmannerandserveasbettersources(incubators),usingaDHSreactor,~2-km-deepsubseafloorcoalbedsamplesobtainedusingtheriser-drillingtechnologyofthedeep-seadrillingvesselChikyuduringtheIntegratedOceanDrillingProgram(IODP),wereporttheextensivemicrobiologicalandbiogeochemicalinvestigationsover1000daysofDHSreactoroperation,includingthedetailedcultivationprocedure,munitystructure,-,anisms,includingamethanogenicarchaeon,(Fig.?1)operationatanearinsitutemperatureof40°Coverthecourseof1029daysyieldedeffluentatthemeanoxidation-reductionpotential(ORP)valueof?430±47mV(n=478),±(SupplementaryFig.?S1).Asanindicatorofmicrobialmetabolicactivity,methaneinthereactorheadspacewasmonitoredcontinu-ouslyduringtheentireperiodofreactoroperationafterthefirstmeasurementat7days(Fig.?1a,SupplementaryTable?S1).Attheearlystage(0–105days),,δ13CandδDvaluesofmeth-anedecreasedsharplyfrom??‰andfrom??‰,respectively(Fig.?2b,c).The13C-depletedmethanesuggeststhatmethaneabsorbedonligniteparticles,originallyestablishedinthesub-seafloorcoalbedlayers,-,continuousmethane-productionwasalsoanicsubstances(.,acetate,propionate,butyrate,andyeastextract)intothemediumafter694days(Fig.?2a).At805days,,lasscolumnowingtotheeffluentpumpfailure(seearrowsinFig.?1),,,,althoughthevaluesfluctuated(Fig.?2b,c).positionδ13CandδDvalueswere??‰,respectively,(DOC)concentrationsweregenerallyhigherintheeffluentthantheinfluent(Fig.?2d,SupplementaryTable?S1),indicatingthereleaseoflignite-,atconcentrationshigherthanwhatwasinitiallyaddedtothebioreactor,??‰(SupplementaryTable?S1),(SupplementaryTable?S1),suggestingtheirutilizationbymicrobialmetabolismwithin70hofhydraulicretentiontime(HRT),butnotbutyrate,|(2019)9:2305|https:///-019-38754-w2:..entificreports/-handsideshowvirginspongecarriers(upper)andlignitecoalparticlesattachedtothespongecarriers(lower).,,suggestingpropionatewasalignite-(SupplementaryTable?S1).anicmatter(DOM)intheeffluentindicatedthattheDOMwasmainlymicrobiallyderived(withafluorescenceindex[FI])andrecentlypro-duced(withabiologicalindex[BIX]of1)(Fig.?3a).Excitation-emissionmatrices(EEM)fluorescencespectros-copyanalysisrevealedthreedistinctphasesofmicrobialactivityassociatedwithlignite-degradationovertheperiodofreactoroperation(Fig.?3).Inthefirstphase(0toapproximately100days),humic--prisedofirreg-pounds,,butotherlignite-derivedstructuressuchasbenzene,(conjugationdegreeofhumic-likesubstances),-poundsadsorbedonligniteand/(100to721days)wascharacterizedbyanoveralllossofhumic-likepeaksandarelativedominationofprotein-pounds(protein-likeoverhumic-like[P/H]>1).-munityand/(821to875days)wasdominatedbyanotableincreaseinhumificationoftheDOM(humificationindex>3),wherehumic-like,conjugated(aro-matic),butareassumedtobe>1000Dainsize29andassumedtocontaineither3to5(Cpeak)or7(Apeak),thehumificationsignalcoincideswiththehighestAC/-pounds,suchasplantligninderivedphenols,(after694days).,-autofluorescentmethanogens,rod-shapedcellsandsphericalspore--riers,whichwerecollectedat694and932days,werealsoobservedbyscanningelectronmicroscopy(SEM).ScientificRepoRts|(2019)9:2305|https:///-019-38754-w3:..entificreports/anicsubstancesNoadditionoftheexternal(acetate,propionate,butyrate,anicsubstancestotheextract)tothemediummedium(a)(days)Methaneconcentration(μM)20100(b)-40-50-60(?vPDB)4-70CHC--8013δ-90-100(c)-200-25