Molecular genetics and diversity of primary biogen

BGD4,349–384,2007MoleculargeneticsofbiogenicaerosolparticlesV.Despr´esetal.TitlePageAbstractIntroductionConclusionsReferencesTablesFiguresJIJIBackCloseFullScreen/EscPrinter-friendlyVersionInteractiveDiscussionEGUBiogeosciencesDiscuss.,4,349–384,2007©Author(s)2007.ThisworkislicensedunderaCreativeCommonsLicense.BiogeosciencesDiscussionsBiogeosciencesDiscussionsistheaccessrevieweddiscussionforumofBiogeosciencesMoleculargeneticsanddiversityofprimarybiogenicaerosolparticlesinurban,rural,andhigh-alpineairV.Despr´es1,3,J.Nowoisky1,3,M.Klose2,R.Conrad2,M.O.Andreae1,andU.P¨oschl11BiogeochemistryDepartment,MaxPlanckInstituteforChemistry,P.O.Box3060,55020Mainz,Germany2BiogeochemistryDepartment,MaxPlanckInstituteforTerrestrialMicrobiology,Karl-von-Frisch-Straße,35043Marburg,Germany3DepartmentofGeneralBotany,JohannesGutenbergUniversity,Saarstraße1,55099Mainz,GermanyReceived:24January2007–Accepted:29January2007–Published:5February2007Correspondenceto:V.Despr´es([e-mail])349BGD4,349–384,2007MoleculargeneticsofbiogenicaerosolparticlesV.Despr´esetal.TitlePageAbstractIntroductionConclusionsReferencesTablesFiguresJIJIBackCloseFullScreen/EscPrinter-friendlyVersionInteractiveDiscussionEGUAbstractThisstudyexplorestheapplicabilityofmolecularmethodsforthecharacterizationofprimarybiogenicaerosol(PBA)particlesintheatmosphere.Samplesoffinepartic-ulatematter(PM2.5)andtotalsuspendedparticulates(TSP)havebeencollectedondifferenttypesoffiltermaterialsaturban,rural,andhigh-alpinelocationsalongan5altitudetransectinthesouthofGermany(Munich,Hohenpeissenberg,Mt.Zugspitze).Fromfilteraliquotsloadedwithaboutonemilligramofairparticulatematter,DNAcouldbeextractedandDNAsequencescouldbedeterminedforbacteria,fungi,plantsandanimals.Sequenceanalyseswereusedtodeterminetheidentityofbiologicalor-ganisms,andterminalrestrictionlengthpolymorphismanalyses(T-RFLP)wereapplied10toestimatediversitiesandrelativeabundancesofbacteria.Investigationsofblankandbackgroundsamplesshowedthatfiltermaterialshavetobedecontaminatedpriortouse,andthatthesamplingandhandlingprocedureshavetobecarefullycontrolledtoavoidartifactsintheanalyses.MassfractionsofDNAinPM2.5werefoundtobearound0.05%inurban,rural,15andhighalpineaerosols.TheaverageconcentrationofDNAdeterminedforurbanairwasontheorderof∼7ngm−3,indicatingthathumanadultsmayinhaleaboutonemicrogramofDNAperday(correspondingto∼105haploidhumangenomes).MostofthebacterialsequencesfoundinPM2.5werefromProteobacteria(42)andsomefromActinobacteria(10)andFirmicutes(1).Thefungalsequenceswerechar-20acteristicforAscomycota(3)andBasidiomycetes(1),whichareknowntoactivelydis-chargesporesintotheatmosphere.Theplantsequencescouldbeattributedtogreenplants(2)andmossspores(2),whileanimalDNAwasfoundonlyforoneunicellulareukaryote(protist).Over80%ofthe53bacterialsequencescouldbematchedwithabout40%ofthe1925T-RFpeaks(58to494basepairlength)foundintheinvestigatedPM2.5samples.TheresultsdemonstratethattheT-RFLPanalysiscoveredmoreofthebacterialdiversitythanthesequenceanalysis.350BGD4,349–384,2007MoleculargeneticsofbiogenicaerosolparticlesV.Despr´esetal.TitlePageAbstractIntroductionConclusionsReferencesTablesFiguresJIJIBackCloseFullScreen/EscPrinter-friendlyVersionInteractiveDiscussionEGUShannon-WeaverindicescalculatedfrombothsequenceandT-RFLPdataindicatethatthebacterialdiversityintheruralsampleswashigherthanintheurbanandalpinesamples.Twoofthebacterialsequences(Gammaproteobacteria)andfiveoftheT-RFpeakswerefoundatallsamplinglocations.1Introduction5BiogenicaerosolsareubiquitousintheEarth’satmosphere,wheretheyinfluenceat-mosphericchemistryandphysics,thebiosphere,climate,andpublichealth.Theyplayanimportantroleinthespreadofbiologicalorganismsandreproductivematerials,andtheycancauseorenhancehuman,animal,andplantdiseases.Moreover,theyin-fluencetheEarth’senergybudgetbyscatteringandabsorbingradiation,andtheycan10initiatetheformationofcloudsdropletsandprecipitationascloudcondensationandicenuclei(Dingle,1966;SchnellandVali,1972;Cox,1995;AndreaeandCrutzen,1997;PruppacherandKlett,1997;HamiltonandLenton,1998;Andreaeetal.,2002;Tay-lorandJonsson,2004;Jaenicke,2005;LohmannandFeichter,2005;P¨oschl,2005a;Duseketal.,2006;McFiggansetal.,2006;SunandAriya,2006).15Primarybiogenicaerosol(PBA)particlesandcomponentsareemitteddirectlyfromthebiospheretotheatmosphere.PBAparticlesrangeinsizefrommillimetersdowntotensofnanometers,andmaythusbemuchsmallerthanoriginallythought(Jaenicke,2005).Particlesofbiologicalorigin,likepollen,bacteria,spores,viruses,plantandan-imalfragments(e.g.,dandruffs,skinfragments),areallwithinthissizerange(Simoneit20andMazurek,1982;Matthias-MaserandJaenicke,1992;Artaxo,1995;Baueretal.,2005).Theactualabundanceandoriginofbiogenicaerosolparticlesandcomponentsare,however,stillpoorlyunderstoodandquantified.ComparedtoconventionalmethodsofPBAanalysis(microscopy,proteinstaining,cultivationofmicroorganisms,etc.),molecularmethodscanprovidemuchmorein-25formation.Theyenabletheidentificationandcharacterizationofbothculturedandunculturedmicro