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3921、、、、。、、、、、。、《》@vip.126.com2015-11-232016－03－094130305841573061TH20144021991-841575181@qq.com*zhangtingxi@njnu.edu.cn。EnvironmentalScience&Technology39920169Vol.39No.9Sept.2016.[J].201639996-102.ZhangDingyuZhangTingxiWangGuoxiang.Developmentandapplicationofsecond-generationsequencingtechnology[J].EnvironmentalScience&Technology201639996-102.1123*1231.2100232.2100233.2100234.210023、。、、。454、SolexaSOLiD、33、、、、、。。X172Adoi10.3969/j.issn.1003-6504.2016.09.0171003-6504(2016)09-0096-07DevelopmentandApplicationofSecond-GenerationSequencingTechnologyZHANGDingyu1ZHANGTingxi123*WANGGuoxiang1231.SchoolofGeographyScienceNanjingNormalUniversityNanjing210023China2.JiangsuCenterforCollaborativeInnovationinGeographicalInformationResourceDevelopmentandApplicationNanjing210023China3.JiangsuProvincialKeyLaboratoryofMaterialsCyclingandPollutionControlNanjing210023China4.JiangsuKeyLaboratoryofEnvironmentalChange&EcologicalConstructionNanjing210023ChinaAbstractThesecond-generationsequencingtechnologybasedonthehigh-throughputsequencingiswiththeadvantagesofhigh-throughputandlowcostcomparedwiththefirst-generationsequencingtechnologybasedonthedideoxychainterminationmethod.Thereforethesecond-generationsequencingtechnologyismorewidelyusedinvariousareas.Thispaperintroducestheworkingprinciplesandmethodsofthesecond-generationsequencingtechnologieswhichinclude454sequencingtechnologySolexasequencingtechnologyandSOLiDsequencingtechnology.Furthermoreapplicationsofsecond-generationsequencingtechnologyinsoilsedimentthehumanbodymarineandsewagearereviewed.Finallytheprospectsofthethird-generationsequencingtechnologyarediscussed.Keywordssecond-generationsequencingtechnologysoilsedimenthumanmarinesewage9[1]。。。1。DNA[2]。、、、、。DNADNA。。1。2、、。。2.11977SangerSanger。DNA。。DNA。。2.2DNA1%。454、ABISolidIlluminaSolexa。2.2.1454454Roche44T、A、C、GPTPPicoTiterPlate。4DNA、ATP、。2454。2.2.2SolexaSolexaIlluminaDNA39FlowcellDNAFlowcellClusterCluster。DNA。3Solexa。2.2.3SolidSolidABI8。DNA。5。4Solid。13。PCRRochePCRQ20400bp0.5GIllumina、、、RNA、PCRQ30100bp30GABISNPGC、SNPPCRQ4050bp50G13Table1Comparisonof3sequencingtechnologies3454。454400bp。Illumina454。。3.1[4-8]1g、。。Buee[9]4546。81%OTUs73%ITS-126OTUAgaricomycetes。。Huang[10]4543AOAAOBsubcluster5subcluste3AOB。subclusters2subclusters9。pHAOAAOB。Ma[11]454O157∶H7、。O157∶H7ECQ201%Q300.1%Q400.01%。9TNWSOC。O157∶H7。。3.2[12-15]。。Wang[16]Illumina、3。。。Wu[17]8。45416srRNA。Jiang[18]illuminaalpha。、、、Deferribacterias。3.3[19-22]。Cooper[23]9730175045414212。。Qin[24]Illumina345。Militello[25]454454。Rutvisuttinunt[26]6MiSeqIllumina。。3.4[27-31]。Goldberg[32]454Sanger。Wilkins[33]4542。。Peressutti[34]4543618846002804770027005。。Sogin[35]4541~2。。Xia[36]、、45439。3.5[37-39]。、3。、。Ye[40]454、、。18808。4AlphaproteoBacteria、Thermotogae、DeltaproteoBacteriaGammaproteoBacteria。。Ye[41]IlluminaHiSeq20002、。AOAAOB2AOBAOA2。QPCRAOAAOB。Ye[42]45416srRNA227458269062。Sanapareddy[43]454DNA0.3%。Lu[44]454、、。、。4Handelsman[45]1998。DNA。。、、16rRNA。45416rRNADGGE、T-RFLPSanger[3]。PCRPCR。1PCR23499.9999％。。。、、。[][1].[J].2007473548-553.LiXiangQinLinDaiShikunetal.Marinemicrobialmetagenomicsprogressandprospect[J].ActaMicrobiologicaSinica2007473548-553.inChinese[2].[J].2008282209-218.HeJizhengZhangLimeiShenJupeietal.Advancesandperspectivesofmetagenomics[J].ActaScientiaeCircumstan－tiae2008282209-218.inChinese[3]CarolaSRolfD.Trendsmetagenomicanalysespastandfu－ture[J].EnvironmentalMicrobiology20117741153-91161.[4]HirschJGalidevaraSStrohmeierSetal.Effectsondi－versityofsoilfungalcommunityandfateofanartificiallyappliedbeauveriabassianastrainassessedthrough454py－rosequencing[J].MicrobialEcology2013663608-620.[5]SchmidtPABalintMGreshakeBetal.Illuminametabarcodingofasoilfungalcommunity[J].SoilBiologyandBiochemistry201365128-132.[6]HagedornFHiltbrunnerDStreitKetal.NineyearsofCO2enrichmentatthealpinetreelinestimulatessoilrespira－tionbutdoesnotaltersoilmicrobialcommunities[J].SoilBi－ologyandBiochemistry201357390-400.[7]FrisliTHaverkampTJakobsenKSetal.Estimationofmetagenomesizeandstructureinanexperimentalsoilmicro－biotafromlowcoveragenext-generationsequencedata[J].JournalofAppliedMicrobiology20131141141-151.[8]McGuireKLPayneSGPalmerMIetal.DiggingtheNewYorkcityskylinesoilfungalcommunitiesingreenroofsandcityparks[J].PlosOne201383e58020.[9]BueeMReichMMuratCetal.454pyrosequencinganalysesofforestsoilsrevealanunexpectedlyhighfungaldi－versity[J].NewPhytologist20091842449-456.[10]HuangLQDongHLWangSetal.Diversityandabun－danceofammonia-oxidizingarchaeaandbacteriaindiverseChinesepaddysoils[J].GeomicrobiologyJournal201331112-22.[11]MaJIbekweAMYangCHetal.Influenceofbacterialcommunitiesbasedon454-pyrosequencingonthesurvivalofEscherichiacoliO157∶H7insoils[J].FemsMicrobiologyEcol－ogy2013843542-554.[12]LecroqBLejzerowiczFBacharDetal.Ultra-deepse－quencingofforaminiferalmicrobarcodesunveilshiddenrich－nessofearlymonothalamouslineagesindeep-seasediments[J].ProceedingoftheNationalAcademyofSciencesoftheUnitedStatesofAmerica20111083213177-13182.[13]SunMYDaffornKAJohnstonELetal.Coresedimentbacteriad