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基于T-RFLP方法的连栽杉木根际土壤细菌群落变化研究.docx

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基于T-RFLP方法的连栽杉木根际土壤细菌群落变化研究 Title:AnalysisofShiftsinBacterialCommunityCompositionintheRhizosphereSoilofSequentiallyPlantedChineseFir,UsingT-RFLPMethod Abstract: Chinesefir(Cunninghamialanceolata)isawidelyplantedconiferoustreespeciesinChina.ContinuousmonocultureofChinesefircanleadtothedeclineoftreegrowthandincreasedsusceptibilitytodiseases.Therhizospheresoil,whichharborsadiversecommunityofmicroorganisms,playsacrucialroleintreehealthandgrowth.Inthisstudy,weaimedtoinvestigatetheshiftsinbacterialcommunitycompositionintherhizospheresoilofsequentiallyplantedChinesefirusingtheTerminal-RestrictionFragmentLengthPolymorphism(T-RFLP)method. Introduction: ContinuousmonocultureofChinesefircanleadtosoildegradation,reducedtreegrowth,andincreasedvulnerabilitytopathogens.Thecompositionanddiversityofmicrobialcommunities,particularlybacteria,havebeenshowntosignificantlyinfluencesoilfertilityandplanthealth.Therhizosphere,thesoildirectlyinfluencedbyplantroots,isahot-spotformicrobialactivity,whererootexudatesandplant-microbeinteractionsshapethemicrobialcommunitystructure. Methods: 1.SampleCollection:RhizospheresoilsampleswerecollectedfromfoursitesthathavebeencontinuouslyplantedwithChinesefirfordifferenttimeperiods(0years,5years,10years,and20years). 2.DNAExtraction:TotalsoilDNAwasextractedusingacommercialDNAextractionkit. 3.PCRAmplification:Thebacterial16SrRNAgenewasamplifiedusinguniversalprimerswithafluorescent-labeledforwardprimer. 4.Terminal-RestrictionFragmentLengthPolymorphism(T-RFLP):TheamplifiedDNAfragmentsweredigestedwitharestrictionenzyme,andtheresultingterminalrestrictionfragments(T-RFs)werelabeledwithafluorescentdye.TheseT-RFsweresubsequentlyseparatedbycapillaryelectrophoresisandanalyzedusingappropriatesoftware. 5.DataAnalysis:TheT-RFLPprofileswereusedtodeterminethebacterialcommunitycompositionandtocomparethesimilarities/differencesbetweendifferentsamples.StatisticalmethodssuchasPrincipalComponentAnalysis(PCA)wereusedtoanalyzethedata. Results: TheT-RFLPanalysisrevealeddistincts