Using Sensitive Plants as Bioindicators of Ground Level ...Using Sensitive Plants as Bioindicators of Ground Level Ozone Pollution Implementation Guide Authors Irene Ladd, Education

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Using Sensitive Plants as Bioindicators of Ground Level Ozone Pollution

Implementation Guide

AuthorsIrene Ladd, Education Co-PI, Surface Ozone Project, NASA Langley Research Center, VA

Susan Sachs, Education Coordinator, Appalachian Highlands Science Learning Center, Great Smoky Mountains National Park, NC

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About the Authors

Dr.IreneLaddhasbeeninvolvedineducationalreformbothlocallyandstatewideforthepast25years.Sheisaretiredteacherof33yearsandservedonNewHampshire’sScienceCurriculumFrameworksCommittee.ShejoinedtheSurfaceOzoneMeasurementforGLOBE(SMOG)Teamtodevelopthesurfaceozoneprotocolandfieldtesttheinstrumentsusedformeasuringsurfaceozone.Shehasdevelopededucationalmaterialstointegratethestudyofsurfaceozonewithcorecurriculum,andthetrainingtoimplementtheprogram.Dr.Laddcontinuestoprovidetrainingnationallyandinternationallyandprovidingguidancetowardtheimplementationofinquiryintheclassroomthroughauthenticscienceandstudentresearch.ThecollaborationtoincorporatetheuseofozonesensitiveplantswiththeSurfaceOzoneProtocolisanaturalextensiontotakingsurfaceozonemeasurementsfortheGLOBE(GlobalLearningandObservationstoBenefittheEnvironment)Program.IrenewaseducatedatKeeneStateCollege,NH,UniversityofNH,andVanderbiltUniversity,TN.

SusanSachsisaNationalParkRangerandtheeducationcoordinatoroftheAppalachianHighlandsScienceLearningCenterlocatedinGreatSmokyMountainsNationalPark.Abigpartofherjobinvolvescreatingedu-cationalopportunitiesfromtheresearchthatoccursintheparksoftheAppalachianHighlandsmonitoringnet-work(besidesGreatSmokyMountainsNationalParkshecoversBigSouthForkNationalRiver&RecreationArea,ObedWildandScenicRiverandtheBlueRidgeParkway).Herpersonaleducationphilosophyisthatpeoplelearnbestwhentheyareinvolvedinlearningactivitiesthatarebothmeaningfulandrelevantsomanyoftheeducationprogramsathercenterinvolvestudents,teachersandothersincollectingdataforactualresearchprojects.Theozonebio-monitoringprojectisoneexampleofseveralcitizenscienceprojectsongoingattheAppalachianHighlandsScienceLearningCenter.SusanwaseducatedattheUniversityofMaryland,CollegeParkandhaslivedandworkedinNationalParksinWashington,DC,Alaska,Arizona,andCalifornia.SusancurrentlyresidesinthebiologicallydiversemountainsofNorthCarolina.

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Acknowledgements

Theinformationinthisguiderepresentscontributionsfromresearchscientists,professionalsmonitoringairqualityandplantresponsesinnationalparksandforests,andotherhighlyregardedsources.Reprintedmaterialisincludedwithpermissionandthesourcesareindicated.ResourcesareidentifiedintheAppendix.

AGLOBE(GlobalLearningandObservationstoBenefittheEnvironment)AdvancedworkshopwasheldattheNorthCarolinaCenterfortheAdvancementofTeachingduringAugust2004.Thebasicatmosphericprotocols,andadvancedsurfaceozoneandaerosolsprotocolswerepartofthetraining.Inaddition,SusanSachs,Educa-tioncoordinatorattheAppalachianHighlandsScienceLearningCenterintheGreatSmokyMountainsNationalParktrainedtheparticipantstoidentifyandestimatethepercentageofozoneinducedfoliarinjurypresentontheCut-leafConeflower.Monitoringsymptomsofozoneinducedfoliarinjurytonativeplantswasanaturalexten-siontotakingsurfaceozonemeasurements.ThroughthecollaborativeeffortsofSusanSachsandIreneLadd,plansweremadetodevelopanimplementationguide,fieldcharts,protocol,andtrainingprogram.Wethankthescientistsfortheircontributionstowardthedevelopmentofthisguide.

Inadditiontodevelopingmaterialstomonitorozoneinjurytonativeplants,wewerealsointerestedinidentify-inganagriculturalcropthatcouldbeeasilyintegratedwithcorecurriculumineducationalandnon-educationalsettings.BillJacksonandAliceCohenfromtheUSForestServiceinNorthCarolinahavecollaboratedwithDr.KentBurkey,USDA-ARSPlantScienceResearchUnitinRaleigh,NC,toconcurrentlyfieldtestozonesensi-tiveandresistantbeans.Theywereusingmodifiedguidelinesandreadilyavailableretailmaterialstomonitorgermination,growth,andozoneinjury.Theresultsoftheparallelstudywillprovidetheinformationneededtodeveloptheprotocolandtrainingprogramformonitoringozonesymptomsonresistantandsensitivebeans.

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Contributing Research ScientistsDr.ArtChappelkaisaProfessorintheSchoolofForestry&WildlifeSciencesatAuburnUniversity,Auburn,AL,USA.HereceivedhisPh.D.inplantpathologyfromVirginiaTechin1986.Since1987,Dr.ChappelkahasbeenonthestaffwithintheSchoolinvestigatingtheresponsesofforesttreesandassociatedplantspeciestoairpollutants.Dr.Chappelkahasauthoredorco-authoredover50peer-reviewedjournalarticlesand12bookchap-ters.Heisareviewerforseveralinternationaljournals,andhasparticipatedonnumerousUSEPApeer-reviewpanels.Hisprimaryinterestsareinairpollutionandglobalclimateeffectstoterrestrialecosystems;nativeplantcommunityresponses(shiftsindiversity)toairpollutantsandglobalclimatechange;plant-stress-airpollution/globalclimatechangeinteractions;airtoxics,andurbanforestry.

Dr.JackFishmanhasbeenstudyingozoneformorethan30years.HecametoNASAin1979wherehehasdevelopedawaytomeasureozonepollutionfromsatellites.Usingsatellites,hediscoveredlargeplumesofozonepollutioncomingfromindustrializedregionsoftheworldsuchastheeasternUnitedStates,Europe,andeasternAsia.Somewhatsurprisingly,healsofoundlargeamountsofozoneovertheSouthAtlanticOcean.Suchplumescouldevenbeidentifiedinmeasurementsthousandsofkilometersfromtheirorigin.In1992,heledagroupofscientiststoinvestigatethecompositionandoriginoftheseelevatedozoneconcentrationswheretheyflewinNASA’sspeciallyinstrumentedDC-8airplaneovera5-weekperiod.ThesourceofthepollutionwasfoundtobewidespreadbiomassburninginsouthernAfricaandBrazilandauniquemeteorologicalsitua-tionthatresultedinbothplumesbeingtransportedoveroceanareasadjacenttoAngolaandNamibia.In1990,Dr.Fishmanco-authoredGlobalAlert:TheOzonePollutionCrisis,abookforgeneralaudiencesthatexpressedhowimportanttheissueofglobalpollutionisandhowwecantakemeasurestoreducethedetrimentaleffectsofwidespreadozonepollution.

Dr.HowieNeufeldreceivedhisB.S.inForestryfromRutgersUniversityin1975,hisM.F.inForestSciencesfromtheYaleSchoolofForestryandEnvironmentalSciencein1977,andhisPh.D.inBotanyfromtheUniver-sityofGeorgiain1984.Hewasapost-doctoralfellowatNewMexicoStateUniversityunderDr.GaryCun-ninghamfrom1984-1985,workingontheecophysiologyofrangegrassesandcreosotebush.In1985hebegananNRCpost-doctoralappointmentunderDrs.DaveTingeyandBillHogsettattheEPALabinCorvallis,OR.Whilethere,heworkedontheeffectsofozoneonrootgrowthoftreeseedlings.Aftertwoyears,hecamebacktotheUniversityofGeorgiaasaresearchcoordinatorintheForestrySchoolbeforeacceptingapositionasAs-sistantProfessorofBiologyatAppalachianStateUniversity.CurrentlyheisProfessorofBiology,Past-Presi-dentofTheAssociationofSoutheasternBiologists(ASB),andPresident-ElectoftheSouthernAppalachianBotanicalSociety.

Dr.Neufeld’sresearchexpertiseisintheareaofplantphysiologicalecology,andhasincludedworkonplantsinswamps,deserts,andforestunderstories.Forthepast20years,hehasbeenactiveinairpollutioneffectsresearch,includingacidicdeposition(rainandfogstudiesonsprucetreesandhardwoodsoftheeasternUnitedStates,andtroposphericozoneonnativewildflowers.From1988-1992hewastheprincipalinvestigatorofaNationalParkService/EnvironmentalProtectionAgency(EPA)sponsoredresearchprojectontheeffectsofozoneonplantsnativetoGreatSmokyMountainsNationalPark.Theseresultshavebeenpublishedinavarietyofjournals,andadditionalpapersareinpreparationandpress.

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Dr.MargaretPippinisaResearchScientistinAtmosphericSciencesatNASALangleyResearchCenter.Mar-garetcametoLangleyin2001aftercompletingpost-doctoralresearchatWesternMichiganUniversity,whereshespecializedinmakingmeasurementsoforganicnitratesattheUniversityofMichiganBiologicalStation.Shecomesfromabackgroundofbothmodelingandfieldmeasurements,withanemphasisondataanalysis.Sheisinterestedintheanalysisofobservationaldatasetstobetterunderstandthechemistryoftheatmospherewithaparticularinterestinthechemistryofbiogenichydrocarbonsandtheirozoneproductionpotential.Mar-garethasbeenactiveinscienceeducationforovertwentyyearsandenjoysworkingwithstudentsofallages.

Dr.PippinhasspentthepastseveralyearsimprovingthequalityoftheGLOBEsurfaceozonemeasurements.Shehasperformedextensivelaboratorytestingtodeterminetherelationshipbetweentheabsorbance(colorchange)oftheEco-Badgetestcardsandtheozoneconcentration,andthedependenceofthiscolorchangeupontemperatureandhumidity.MargaretisalsoinvolvedinGLOBEteachertrainingfortheSurfaceOzoneProtocolandoftenvisitsGLOBEschoolstodiscussatmosphericscienceandmethodsofanalysiswiththestudents.

Dr.JohnM.SkellyisRetired’04,ProfessorEmeritusPlantPathologyfromtheDepartmentofPlantPathology,PennsylvaniaStateUniversity.BSForestry’62;MS’64andPh.D’68,PlantPathologyPennState.HetaughtatVirginiaTech1968-82;andreturnedtoPennState1982-2004.Hisexpertiseinforestpathologywithspecial-izationinairpollutioncausedeffectstoforesttreesandnativeplantswithinNorthTemperateregions.JohnhasbeeninvolvedinwritingtheCriteriaDocumentsfortheUSE.P.A.andasaconsultanttotheU.S.Dept.OfJus-ticeandtheUnitedNationsondiagnosingairpollutioncausedinjurytoplants.Internationalcooperativeproj-ectshavebeenheldwithcolleaguesinCanada,Mexico,Switzerland,Spain,Germany,andItaly.

Contributing Research Scientists

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Organization of this guideThisguideisdesignedasaresourceandfieldmanualtoimplementtheobservationandrecordingofozonein-ducedfoliarinjurytoCut-leafconeflower(Rudbeckialaciniata)andCommonmilkweed(Asclepiassyriaca).

Section Iprovidesbackgroundinformationontheformationofozoneairpollutionandtheimportanceofunderstandingthetransportpatternsoflocal,regionalandgloballevelsofozoneairpollutionanditsimpactonvegetationasitistransportedfromurbanindustrialcenterstomoreruralandforesteddownwindregions.

Section IIprovidesageneralintroductiontothecharacteristicsofozone’sinducedfoliarinjurytotheleavesofbroadleafspeciesandtheprocessesusedtoidentifyozonesensitiveplants.Theguidealsodescribesthescien-tificprocessesusedtoidentifyozone-inducedfoliarsymptomsonplantsintheenvironmentandincontrolledopentopchambers.

Section IIIdescribesmethodstobeusedinsettingupabioindicatorgardenfordetectingozone-inducedinju-ries,andtheprotocolforgatheringandsubmittingdataonCut-leafconeflower(Rudbeckialacinata),andCom-monmilkweed(Asclepiassyriaca)totheHandsontheLandWebsite,hosttotheinteractivedatabase.

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Using Sensitive Plants as Bioindicators of Ozone Air Pollution

Table of Contents

SectionI: FormationofSurfaceOzoneAirPollution IntroductiontoOzone QualityofAir OzonePollutionCycle InfluenceofWeatheronOzoneFormationandEpisodicEvents

SectionII: InvestigatingFoliarInjuryonOzoneSensitivePlants OzoneAirPollutionisHarmfultoHumans StudiesofOzone-sensitivePlants FumigationStudies

SectionIII: OzoneBioindicatorGarden BackgroundInformation OzoneMeasurementsandPlantObservations Planningthe“Ozone”Garden OrganizingthePlantsforStudy PreparingtoObserve TrainingtoAssessFoliarInjury RegisteringYourSite EnteringData RetrievingData LookingAtData AretheDatareasonable? WhatDoPeopleLookforinTheseData AnExampleofaStudentResearchInvestigation IntroductiontoLessonPlans LeafGame O3SkillsWorksheet RatingOzone-InducedFoliarInjuryonOzone-sensitivePlants SiteDefinition SiteDefinitionWorksheet FieldInstruction RatingOzoneInjury InjuryGuideforMilkweed DataSheets OzoneGardenFieldGuide WebSites References

888810

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191919202021222222232424252730313234363738384243485456

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Introduction to Ozone Formation

QualityofAir Theatmosphereismadeupofamixtureofgases.Thegasesincludenitrogen,oxygen,carbondioxide,watervapor,argon,andtracegases.Nitrogenmakesupapproximately78%oftheatmosphereandoxygen21percent.Onlyabout1%oftheatmosphereismadeupofacombinationoftheremaininggases. Thequalityofairisaffectedbythecompositionoftheatmosphere.Theadditionofpollutantscreatedfromvarioushumanactivitiesgraduallycontributestothedeteriorationofairquality.Thestudyofsurfaceozone(ozoneproducednearthegroundwherepeoplebreathetheair)andusingozonesensitiveplantsasbioindicatorsofozoneinducedplantinjurywillhelpdevelopanunderstandingofhowhumanbehavioraf-fectsthequalityofairandanawarenessofitsenviron-mentalimpact. Ozonegasisaformofoxygen.Itisacolorless,highlyreactivegasthatexistsfromtheEarth’ssurfacemilesupintotheatmosphere.Ozonehasthreeatomsofoxygen(O3),whereastheoxygenmoleculeweneedtobreatheonlyhastwoatoms.Itisthethirdatomthatmakesozoneahighlyreactivegas.

“Good”ozoneislocatedintheleveloftheatmo-spherecalledthestratosphere.IttrapsultravioletraysandprotectsthelifeformsonEarth.Thisistheozonelayerwehearsomuchabout.Incon-trast,ozoneproducedinthetroposphereatthelevelwebreatheisconsideredapollutantandharmfultoplantsandanimals.Ozoneatthisleveliscalledsur-faceozone,or“bad”ozone.Itisthemaincomponentofsmog. Thelowconcentrationsofsurfaceozonethatnatu-rallyoccurinthetroposphereareproducedbyin-tensesunlightdrivenreactionsinvolvingseveralkeyhydrocarbonsgivenoffbyplantsandnitrogenoxidesproducedaspartofthenaturalnitrogencyclesofEarth’satmosphere.Undernormalconditions,thesephotochemicallydrivengasesmayproduceozoneconcentrationsbetween10-40ppbdependingupontheseasonoftheyear(Lefohnetat.1990). Othernaturalsourcesofozonemayincludelight-ningandturbulentweatherconditionsthattransportconcentrationsofozonedowntoEarth’ssurfacefromthestratosphere.Thisphenomenonoccurscom-monlyonmountaintopsduringthunderstormsandhasbeenlinkedtosymptomsoneasternwhitepineduringlatespringandearlystormsthroughouttheAppalachianMountains.

OzonePollutionCycle

Humanactivityhasaddednewconcentrationsofpollutantstotheairwebreathe.Themajorcontribu-torsarevehicles(automobiles,trucks,busesandairplanes)andindustrialburningoffossilfuels(asoilandcoalburningutilityplants).Theprimarygasesthatcontributetotheproductionofsurfaceozoneareexhaustsfromanyinternalcombustiongasoline-fu-eledengine.Themajorby-productsofthecombus-tionprocessare:carbonmonoxide(CO),nitrogenoxides(NO),andpartiallyburnedhydrocarbonscalledvolatileorganiccompounds(VOCs)thatareveryreactive. Nitrogenmoleculesintheatmospherearenon-re-activetothesun’senergy(theydon’tsplitintoatomsandattachtootheratomstoformsomethingnew),buttheintenseheatwithinthecombustionchamber

SECTION I: Formation of Ozone Air Pollution

ImagecourtesyoftheNationalParkService

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causesthenitrogenmoleculestosplitintotwonitro-genatoms.Thesesplitnitrogenatomslinkupwithoxygenatomsandformabyproductcallednitrogenoxide(NO). Theairtakenintotheenginecombineswiththegasoline(hydrocarbons)beforeenteringthecombus-tioncylinders.Iftherewerecompletecombustion,asthehydrocarbonorgasmoleculesigniteandsplitandproduceenergyintheengine,thebyproductswouldbecarbondioxide(CO2)andwatervapor.However,completecombustiondoesnotoccurandsomehydro-carbonsremainasVOCs.

Hydrocarbonsarealsoabyproductwheneverthingsareburned.Forexample,aforestfireproduceshydro-carbons,whicharemoleculesofcarbonandhydrogen.Butthesearefar-lessreactiveandoflesserimportancetotheformationozonethanaretheVOC’semittedwithinindustrialandautotransportationexhausts.Thepiegraphbelowidentifiesthemajorsourcesofprima-ryairpollutantscontributingtotheproductionofthesecondarypollutant,surfaceozone.

Thesenewgaseshavecontributedtotheproduc-tionofhigherconcentrationsofsurfaceozone,i.e.theformationofozoneairpollution.Ingeneral,ozoneisamaincomponentofurbansmog,anditiscommonly

CourtesyoftheEnvironmentalProtectionAgency

referredtoasphotochemicalsmog.Theproductionofsurfaceozoneairpollutionusuallypeaksinthelaterafternoonatlowelevations,butmayremainhighalldayathigherelevations,suchasinthemountains.

Theafternoonpeakatlowelevationsoccursbe-causetheformationofozoneisdependentonthesun’energy,anditproceedsfasterathighertemperatures,sotheconcentrationstendtobuildupfrommorningtotheafternoon.Atnightthough,otherchemicalsintheatmospheremaybreakdowntheozone,whichiswhyitstartsoffeachdayatalowerconcentrationthantheprecedingafternoon. Duringthecourseoftheday,thepollutantsreactinthepresenceofthesun’sradiantenergyandtheoxidesofnitrogen(NO)produceconcentrationsofozone,airpollution.Aphotochemicallyformedhazyairmassisformedthathasabrowntingeduetocombustioninfuelpoweredengines.

Whethertheprimarypollutantsareproducedthroughaforestfireorthefireofacombustionengine,thebyproductsproducedaretheprecursorsneededfortheproductionofsurfaceozone.Theseprimary

http://www.york.ac.uk/inst/sei/APS/a-proj.htm

http://www.epa.qld.gov.au/environmental_management/air/air_quality_monitor-ing/air_pollutants/ozone/

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pollutantsreactwiththeenergyofthesuntoproduceasecondarypollutant,surfaceozone. SurfaceconcentrationsofozoneairpollutionbegintoincreaseseasonallyfromAprilthroughSeptemberintheeasternregionsoftheUnitedStates.Thisisthetimeperiodofincreasedamountsofsunlight,highertemperatures,andcommonlyoccurringstagnatinghigh-pressuresystems(BurmudaHighs)overvastregionsoftheMidwestandMid-AtlanticStates.Un-dertheseatmosphericconditionsozoneairpollutionreachesitshighestlevelsduringthehottestandmostsunlitmonthsoftheyear.Dependinguponweatherpatterns,theconcentrationsofozoneairpollutionthoughusuallyhigherinthesummer,canvaryyeartoyear.Forexample,duringthehot,dryyearsofthemid-tolate1990s,ozonereachedrecordlevelsintheGreatSmokyMountainsNationalPark,butin2004,awetandcoolyear,ozonelevelsweregreatlyreduced.Theamountofozoneformedeachyearishighlydependentonweatherconditions,andlesssoonvariationsinpollutantemission,whicharerelativelyconstantfromyeartoyear.

ThetwosidebysidephotosaboveshowthehazeprobleminGreatSmokyMountainsNationalPark.Theweatherconditionsofhot,humid,stagnantairthatcausesulfurparticlestocombinewithwatervaportocreatehaze,arethesameconditionsthatincreasethegroundlevelozonelevels. Initiallysurfaceozonewasthoughttobealocalproblem,butthepollution-ladenairmassesmaytravelbeyondthelocalareaswheretheywereoriginallypro-duced.Theslowmovingairmassespickuppollutants

Influence of Weather and Episodic Events on Ozone Formation

alldaylongastheytraveloverindustrialcenters;largefossilfuelfiredpowerplants,incinerators,andmostimportantlyoverlargeandevensmallurbanareaswithmanyformsoffossilfuelfiredtransportation.Thecollectionofpollutantgasesinslow-movingairmassesquicklyreactsinthewarmsunlitair.Themostimportantairpollutantformedwithintheseairmassesisozoneairpollution. Inthe1950sand1960s,theLosAngelesareawastheregionmostaffectedbytheproductionofphoto-chemicalsmog.AfterthepassageoftheCleanAirActin1970,manymoremonitoringstationsweresetupacrosstheUnitedStatesandnowwecommonlyseeincreasedpollutionwellbeyondthelocalareaswheretheywereinitiallyproduced.Slowmovingairmassesallowpollutantsto“cook”alldaylong.Astheymeander,evenmoreozonecanbeformedastheypassoverevenrelativelysmallurbanareas.Complicatedchemistryandmeteorologycanresultinthetransportofsurfaceozonepollutiontoareasthataregenerallyconsideredruralorevenpristine,suchasNationalParks.

TheabovefigureforJuly13,1995,showsthesketchyhighozonepatternswheredetrimentalcon-centrationsarefoundthroughoutruralareasinIn-diana,OhioandPennsylvaniaaswellasindenselyforestedregionsinTennessee,Georgia,andNorthCarolina.Dependingonthemeteorologicalsituation,highlevelsofozonecanremainoveranareafora

ImagecourtesyofEnvironmentalProtectionAgency’sAirNowwebsite.

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periodoftimeandplantswillbeexposedtoozoneairpollution. Slowmovinghigh-pressureairmassesarecommonoccurrencesintheeasternUnitedStates.Theseairmassestransportsignificantconcentrationsofozoneairpollutiontotheplantcommunitiesofmountainous,forested,andruralareasdownwindoftheindustrialurbanareas.

ThemapaboveisasatelliteimagethatidentifiesthelocationofindustrialemissionsintheU.S..Duetothecontinuingphotochemicallydrivenprocessesastheairmassslowlymovesdownwind,thesemoreremoteareasoftenhavegreaterozoneexposuresforlongerperiodsoftime.Ifthehighairpressuremassstag-natesandremainsoveranareaforaperiodoftime,

LocationofIndustrialEmissionsinU.S.derivedfromasatellite.Itistheclimatologyfortheyear2003.

thepollutioncanbuildupandexposeplantstomoreseverelevelsofozone. Althoughsomecontrolshavebeenimplementedthathavehelpedtodecreasethelevelofprimarypol-lutantgasesenteringtheatmospherethatcontributetotheproductionofozoneairpollution,thelargeincreaseinthenumbersofvehiclesandtheincreaseinthemilesdrivenhaveoffsetmanyofthegainsthathavebeenmadethroughtheimplementationofpol-lutioncontrols.NewsatelliteinstrumentscannowmapozonepollutionandtheglobaldepictionaboveshowsthathighozoneconcentrationsarenotuniquetotheeasternU.S.orCalifornia.Inthesummertime,evenhigherconcentrationsarenowfoundoverChinaandIndia,wherepollutioncontrolsarenotasrigidasintheU.S.Furthermore,scientistsalsobelievethatemissionsfromtheseregionswillcontinuetogrowasthesecountriescontinuetodevelop.

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SECTION II: Investigation of Ozone Injury Induced Foliar Injury to PlantsSurface Ozone is Harmful

OzoneAirPollutionisHarmfultoHumans Highconcentrationsofozoneairpollutionareharmfultobothanimalsandplants.Concernforairqualityisreflectedmostduringhotsummerdaysas“ozonealertdays”orsomeotherphraseisusedtowarnchildrenandtheelderlytorestrictoutsideactiv-ityduetothelikelihoodofhigherexposurestoozone.Why?Becauseexposuretoelevatedconcentrationsofsurfaceozoneoverextendedperiodsoftimecauseshealthproblems.Itdamagestheimmunesystem’sdefensesmakingonesusceptibletolunginfections.Ozonecausesacuterespiratoryirritation,breathingproblems,andaggravatesasthma.Thepollutantde-creaseslungcapacityby15%toover20%.Thisisbe-causeozone“sunburns”yourlungs.Ozonedamagesthecellsthatlinetheairspacesinthelung. Withinafewdays,thedamagedcellsarereplacedandtheoldcellsareshed-muchinthewaythatskinpeelsafterasunburn.Ifthiskindofdamageoccursrepeat-edly,thelungmaychangepermanentlyinawaythatcouldcauselong-termhealtheffectsandalowerqual-ityoflife.Childrenareatgreatestriskfromsurfaceozoneexposure,becausetheyspendmoretimeout-sideinvolvedinvigorousactivitiesandhaveagreaterdemandforintakeofair.Theirrespiratorysystemsaredevelopingandaremostsusceptibletopermanentdamage.Theelderlyarealsomoresensitivetozonebecausetheirimmunesystemisnotasgoodasitusedtobe.Formoreinformation,visittheEPAwebsiteandviewtheirpublicationcalled“WhatYouNeedtoKnowAboutOzoneandYourHealth”athttp://www.airnow.gov/index.cfm?action=health2.smog1#3

Thephotosaboveshowahealthylungairway(left)andaninflamedlungairway(right).Ozonecaninflamethelung’slin-ing,andrepeatedepisodesofinflammationmaycauseperma-nentchangesinthelung.(EPAphotosfrom“WhatYouNeedtoKnowAboutOzoneandYourHealth”)

OzoneAirPollutionisHarmfultoPlants. Plantsarenotunlikehumans.Theyneedtotakeincarbondioxideforphotosynthesis,theprocesstheyusetoproducetheirownfood,andgiveoffoxygenasabyproduct.Thehealthyplants,likeyoungchildren,areactively…”taking”…intheair. Ozoneentersleavesthroughtheirstomata.Stomataaresmallporesintheleafthatallowgasestoenterorleave.Whentheyareopen,carbondioxidecanenter,whichtheplantusestomakeitsfood,suchassugarsandstarches.Atthesametime,waterexitstheleaf,throughtheprocessknownastranspiration.Ifozoneispresentintheair,ittoowillentertheleafthroughtheopenstomata,followingmuchthesamepathasthecarbondioxide.

Ozone,however,isahighlyreactivemolecule,andonceitenterstheleaf,itwillfinditswaythroughtheleafinteriortothecellsresponsibleforphotosynthesis,particularlythepalisadeandspongymesophylltis-sues.

http://res2.agr.gc.ca/publications/ha/3d_e.htm

Cuticle-waxlayer

Epidermis-clearcells

Mesophypll-Palisade

Mesophypll-Spongy

Epidermis-clearcells

Cuticle-waxlayer

Crosssectionviewofaleaf,sourceUSForestService

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Thesurfacelayerofcellsinaleafarecalledtheepi-dermis.Thelongcellsbelowarethepalisademeso-phyllcells,wheremuchofthephotosynthesisoccursinleaves.Theirregularcellsfurtherdownarespongymesophyllcells,andtheyalsoparticipateinphoto-synthesis.Atthebottomisthelowerepidermallayer,andthecurvedcellsrepresentguardcellsforstomatawheretheozoneentersontheundersideoftheleaf.Assoonastheozoneenters,itmostlikelyreactswithmoleculesinthecellwallthatenduptriggeringpro-ductionoftheROSmolecules,whichdamagethecell.Ozoneitselfrarelygetsfarintoacelltocausedam-age.Notetheenhancedairspacesbetweenthespongymesophyllcellscomparedtothatinthepalisadelayer.Thisiswherephotosynthesisoccursintheleaf.

Itisofinterestthateventhoughozoneprimarilycomesthroughthelowerepidermalstomata,itistheupperepidermallayer,andeventuallythepalisadecellsthatsufferinjuryfirst,duetotheirinteractionwithlight,inreactionsnotyetfullyunderstood.Ithasbeennoticedthataleafpartiallyshadedbytheleafabovemayonlyshowinjuryintheareaoftheleaffullyexposed,theshadedpartoftheleafshowsnoinjury. Verylittleozonecangetintoaleafthroughthecu-ticle,awaxylayerthatcoversmostofthesurfacesofleavesandstems.Thiswaxylayerisnearlyimperme-abletowater,andalsotomostgases,includingbothcarbondioxideandozone.Therefore,theuptakeofozonedependsnearlyentirelyonwhetherthestomataareopenorclosed. Stomataopeninresponsetocertainenvironmental

CO2

O2

Cuticle-waxlayer

Epidermis-clearcells

Mesophypll-Palisade

Mesophypll-Spongy

stimuli.Theseincludelight,highhumidity,andhightemperatures.Atnight,stomatatendtoclosebecauseofthelackoflight.Thusozoneuptakeisgreatlyreducedatthistimeoftheday.Ifthehumidityshouldgodown,thiscausesexcessivewaterlossfromthecellssurroundingthestomatalpore,andthestomataclose.Withrespecttotemperature,stomatagenerallyopenmorethewarmeritgets.Finally,highcarbondioxidelevelswillalsoclosestomata.

Astomateiscomprisedoftwoguardcells,andsomeaccessorycellsthatassistinopeningandclos-ing.Whentheguardcellsfillwithwater,theypullawayfromeachother,andthestomatalporeisopened.Whentheylosewater,thetwoguardcellscollapseagainsteachother,closingthepore.Thefigureaboveshowsaclosedporeontheleft,andanopenoneontheright. Allplantcellsaresurroundedbyacellwall,com-posedmainlyofcelluloseandlignins.Thesefairlyrigidmoleculesprovidesupportfortheplant.Inorderforozonetodamageacell,itmustgothroughthecellwallfirst,beforereachingthecell’smembrane.Thecellwallmaycontainanti-oxidants,whicharemole-culesthatcanreactwith,anddetoxify,theozone.Thisisonewayfortheplanttoprotectitselffrompatho-gensandtoavoidproblemsfromozone. Onesuchanti-oxidantisVitaminC(alsoknownasascorbicacid)whichisproducedbyplants.IfthereisalotofVitaminCinthecellwall,itcandestroytheozonebeforeitreachesthecellmembrane.Ifnot,thentheozonecanreachthecellmembrane,whereitcancausedamage,eventuallykillingthecell.symptomsthanwell-wateredplants. Ifaplantissufferingfromdroughtstress,itwillcloseitsstomatatosaveonwater.Whilethismaypreventtheplantfromdryingout,italsolowersphoto-synthesis,sincethecarbondioxidecannotgetintotheleaf.However,onebenefitisthatozonealsocannot

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getintotheleaf.Thus,plantsunderdroughtstressoftenshowfewersymptomsthanwell-wateredplants.Whenozoneistakenintoaleaf,itcaninterferewithaplant’sabilitytoproduceandstorefood.Itweakenstheplantmakingitlessresistanttodiseaseandinsectinfestations.Insomesensitiveagriculturalcropssuchasvarietiesofsoybeansandsnapbean,exposuretoozoneairpollutionalsoaffectstheplant’sabilitytoreproduce,thusdecreasingcropyield.Someplantsaremoresensitivetosurfaceozonethanothers,andshowvisiblesymptomswithindaysorweeksoftheirexpo-sureunderfieldandforestconditions. Ozonecausesveryspecificanduniquesymptomsonbroadleafplantsinthefield.Themostcommonsymptomisstippling(alsocalledpurpling)ontheuppersideoftheleaf’ssurfacewiththelowerleafsurfaceabsentofsymptoms.Stipplingconsistsofverysmallspotsmuchlikesomeoneshookfinepepperontotheleafsurface.Thecolorofthestipplingdependsonthespeciesofplant.Theamountofstipplingcan

varydependingupontheenvironment(nutrientsinthesoil,amountofwater,amountofsunlightthatarelocalweatherconditions),andthephysiologicalconditionsormakeupoftheplant.Thestipplingmayoccuronlywithincertainareasoftheleaf’ssurface,butdoesnotinvolveanyofthelargeorsmallveinsintheleaf.Thatis,withozone-inducedstippling,theveinsarefreeofanysymptomswhereasmanyinsectswhilefeedingandcertainfunguspathogenscausedirectinjuriestotheveins. Ingeneral,thelowerolderleavesontheplantex-hibitmorestipplingthanthenewerleavesontheplant.Plantsgrowfromthetipsoftheirstemsandhencethenewleavesthatwereproducedfirstinthespringareatthebaseofthenewshootandarethereforeexposedtotheozoneairpollutionasitoccursthroughouttheen-tirespringandsummerseason.Newleavesproducedlaterinthesummer“see”farlessozonepollutionbecausetheyhavenotbeenpresentallseasonlong.

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Studies of Ozone-Sensitive Plants

ManystudiesandrecentlypublishedreportsintheUnitedStateshavedocumentedozoneinducedfoliarin-juryonozonesensitiveplantspecies.Theinjuriesareeasilyobservedonsensitiveforesttreespeciessuchasblackcherry(Prunusserotina,Ehrh.),whiteash(Fraxinusamericana,L),andYellow-poplar(Liriodendrontulipifera,L.)(Chappelkaet.al.,1992;Skellyetal.,1996;Neufeldetal.,1992;Hildebrandeetal.,1996).Long-terminvestigationsofselectedplantshavebeenconductedinopen-topchambersinTheGreatSmokyMountainsNationalParkofTennesseeandNorthCarolina,theShenandoahNationalParkinVirginia,andtheAlleghenyMountainsofnorth-centralPennsylvania.Afewsamplesofozonesensitivenativespeciesarelistedbelow. 1.Acerrubrum(Redmaple)2.Aesculusoctandra(Yellowbuckeye)3.Apocynumcannibinum(Indianhemp)4.Asclepiasexaltata(Tallmilkweed)5.Asteracuminata(Whorled-woodaster)6.Asterdivaricatus(White-woodaster)7.Asterpuniceus(Purple-stemmedaster)8.Betulalutea(Yellowbirch)9.Cacaliarugelia(Rugel’sragwort)10.Cerciscanadensis(Easternredbud)11.Cornusflorida(Floweringdogwood)12.Eupatoriumrugosum(Whitesnakeroot)13.Glycerianubigena(SmokyMtn.mannagrass)14.Krigiamontana(Dwarfdandelion)15.Liquidambarstyraciflua(Sweetgum)16.Liriodendrontulipfera(YellowpoplarorTuliptree)17.Lobeliacardinalis(Cardinalflower)18.Magnoliatripetal(Umbrella-leafmagnolia)19.Oxydendronarboretum(Sourwood)20.Pinuspungens(Pitchpine)21.Pinusvirginiana(Virginiapine)22.Platanusoccidentalis(Americansycamore)23.Prunusserotina(Blackcherry)24.Phuscoppalina(Wingedsumac)25.Robiniapseudoacacia(Blacklocust)26.Rudbeckiahirta(Black-eyedSusan)27.Rudbeckialaciniata(Cut-leafconeflower)28.Rubuscanadensis(Thornlessblackberry)29.Rubusidaeus(Redraspberry)30.Sassafrasalbidum(Sassafras)31.Vernonianoveboracensis(NewYorkironweed)

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Using Ozone Sensitive Plants as Bioindicators of Ozone InjuryTheuseofbiologicalindicatorstodetectthepresenceofozoneinjurytoplantsisalongstandingandeffec-tivemethodology.Abioindicatorcanbedefinedasavascularornon-vascularplantexhibitingatypicalandverifiableresponsewhenexposedtoaplantstresssuchasozone.Tobeconsideredagoodindicatorspeciesplantsmust:1)exhibitadistinct,verifiedresponse,2)havefewornoconfoundingdiseaseorpestproblems,and3)exhibitgeneticstability. Thesesensitiveplantscanbeusedtodetectthepresenceofozoneataspecificlocationorregion,andprovideuniqueinformationregardingambientairqualityinaparticulararea.Plantindicatorsofozoneinjurycanbeeitherintroduced(BelW3tobaccoforexample)ornative(CommonmilkweedandCut-leafconeflower).Indicatorspeciesaremosteffectivewhenrelatedtoambientozoneconditions.

Fumigation Experiments

Commonmilkweed(Asclepiassyriaca)CommonmilkweedisanativeperennialfoundthroughouttheeasternhalfoftheUnitedStates,ex-ceptalongsomeareasoftheGulfCoast.Thespeciesspreadsnaturallybyseedandfromlongundergroundrhizomes.DuringtheozoneseasonintheBlueRidgeMountainsofVirginia,Commonmilkweedswereobservedtodeveloppurplestippling(adiscreteandveryfinepurplecoloredspotsoftheupperleafsur-facesoftheolderleaves)andeventualoverallchlo-rosis(theyellowingoftheleafcausedbythelossofchlorophyllneededforphotosynthesis).Thequestionraisedwas:Issurfaceozoneresponsibleforthefoliar(plant)injury?Astudywasdevelopedtodeterminewhetherlong-rangetransportedozoneairpollutionintototheBlueRidgeMountainsandtheShenan-doahNationalParkwasresponsiblefortheobservedsymptoms.Eleven-week-oldmilkweedplantswererandomlyplacedintochambersandexposedtooneofthreefollowingozonelevels:0.00ppb,50ppb,and150ppbforsixhoursadayforsevendays.Withinthefumigationchambersthetemperaturerangedfrom20o-27oCandtherelativehumiditybetween70%-80%.Foliarsymptomsdevelopedontheupperleafsurfacesofmilkweedplantsinthechamberswithin2-5daysaftertheywerefumigatedwith(exposedto)thediffer-entlevelsofozone.

Themilkweedplantsinthechamberswiththehigherozoneexposuresdemonstratedincreasedstipplingontheleaves.Bymid-June,stipplingandchlorosiswereobservedonmilkweedplantsgrowinginnofilteredairinopen-topchambers,aswellasonthemilkweedgrowinginopenplotsinthefield.(DuchelleandSkelly,etal.1981).Milkweedsgrowingincharcoal-filteredairsuppliedchamberswerefreeofsymptomsandconsiderablylargerandgreenerthanthoseintheopenplotsandnon-filteredairsuppliedchambers.ThesetwoinitialstudiesconfirmedthatCommonmilkweedwasverysensitivetoozoneairpollutionwithinthepollutedairmassesbeingcarriedfromlong-distances(theMidwestandOhioValley,USA)intotheBlueRidgeMountainsofVirginia.Thefollowingpicturesshowmilkweedplantsthatarehealthyandinjuredasaresultofexposuretoground-levelozone.(NPSPhotos)

OzonechamberusedinGreatSmokyMountainsNationalPark

Tallmilkweedwithnoozoneinjury.

HeavilyinjuredTallmilkweed

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Thechloroticsymptoms(theyellow)intheleavesareastressresponsetotheplant’slong-termexposuretoozoneairpollution.Asthechloroticsymptomsin-creasetheclosertheleafistosenescenceordeath.

Open-top Chamber Studies in Great Smoky Moun-tains National ParkInadditiontomilkweed,scientistsdidexperimentstofindotherplantssensitivetoexposuretosurfaceozone.In1987,astudywassetupintheopen-topchambersattheTwinCreeksResearchfacilityinGreatSmokyMountainsNationalPark.From1988to1992,nearly40differentspecieswerefumigatedwithozoneofdifferingconcentrations,rangingfromcharcoal-filteredchambers(whichscruboutmostoftheozone)toambient(sameasintheairoutsidethechambers),to50%and100%aboveambientlevels(Neufeldetal.1992). Avarietyofspecies,includingbothtreesandwild-flowers,wereexposedtoozoneoverthecourseofaseason.Someperennials,andtrees,wereexposedtoozonefortwogrowingseasons.Theaimofthesestudieswastoverifythattheozone-likesymptomsseeninthefieldonthesespecieswas,indeed,duetoozone.Ofthe30speciesthatwereabletobegrownandfumigatedinthechambers,27showedozone-likesymptomsafterbeingexposedtoelevatedozone.Thus,theresearchersweremoreconfidentthatthesymptomstheywereidentifyinginthefieldwereduetoozoneandnototherfactors. Formostspeciesthatweresensitivetoozone,growthwasreducedwhenevertheozonewaselevatedaboveambient(the50%and100%treatments).Cut-leafconeflowerswereexposedovertwoconsecutive

growingseasons.Afterthefirstseason,therewerenoeffectsoneitherleaforflowerweights,butozone-inducedfoliarstipplewasgreatlyaccentuatedinthehighertreatments.Althoughtheresearcherscouldnotdetectanygrowtheffects,itwasapparentthattheleaveswerehighlysensitivetoelevatedozone.Forotherspecies,suchasBlackcherry,Yellowpoplar,andWhorled-woodaster,thereweresignificantgrowthreductionsintheelevatedozonetreatments.

Cut-Leaf Coneflower (Rudbeckia laciniata)

Cut-leafconeflower(Rudbeckialaciniata)isanativeperennialfoundthroughoutmostoftheUnitedStates.Itgrowsnaturallybyseedandfromlongspreadingrhizomes.Cut-leafconeflowerisever-presentintheGreatSmokyMountainsNationalPark,foundeleva-tionsrangingfrom2,000to6,600feet.Itgrowswellwithintheenvironmentoftheopen-topchambers.Aspartofalong-terminvestigationoftheecologyintheGreatSmokyMountainsNationalPark,Cut-leafconeflowerhasbeenevaluatedforozone-inducedfoliarinjury.

FieldevaluationsweredoneinplotslocatedintheGreatSmokyMountainsNationalPark,whichisinthesouthernAppalachianMountainsofwesternNorthCarolinaandeasternTennessee.Cut-leafconeflowerwasevaluatedonClingmansDometrail,Tennessee,fromJunetoSeptemberandatPurchaseKnobnearWaynesville,NCduringJuly.Theplantssampledwereabout1meterfromeachother,andcategorizedasnearandoff-trailplants.Neartrailplantswerelo-

Yellowpoplarwithozoneinjury,photocourtesyofUSForestService.

Cut-leafConeflowershowingozonedamage.NPSphoto

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catedlessthan5metersfromthetrail.Offtrailplantsweremorethat5metersfromthetrail.Therewerefoursamplingperiodsforozoneinjuryatapproximate-ly3-5weekintervalsfromJunetoSeptemberduring2000.Cut-leafconeflowerswerealsoexaminedonandoff-trailatPurchaseKnobintheGreatSmokyMountainsnearWaynesville,NCduringJuly.Thelevelofozone-inducedinjurywasassessedusingtwomethods:thenumberofinjuredleavesandthepercentageofleafareainjured.ThefindingsofthestudyatClingmansDometrailidentified50%oftheoverallpopulationoftheplantsstudiedhadozoneairpollutioninjury.Plantsgrowingnearthetrailhadsig-nificantlygreaterinjuryand3.5timesgreaterleafareainjurythanthosegrowingoff-trail.Theleavesonthelowerhalfoftheplanthad95%oftheleafinjury.Thepatternofinjurywassimilarforplantsnearandoff-trail.Theresultswithonandoff-trailatPurchaseKnobshowednodifferencesinthelevelofinjurybe-tweentheplants.However,ozoneinjurywasgreaterfortheClingmansDomeplantsthanthoseatPurchaseKnob.(Chappelka,Neufeld,Davison,Somers,andRenfroetal.2003). Ground-levelozonehasbeenidentifiedasthemajorpollutantcausingfoliarinjurytoplants.Thetwonativeozone-sensitiveplantsthatcaneasilybeincor-poratedintoa“BioindicatorGardenfortheDetectionofOzoneAirPollution”andusedasindicatorsofthepresenceofozoneareCut-leafconeflower(Rudbeckialacinata)andCommonmilkweed(Asclepiassyriaca).Theyareperennialsthatarerelativelyeasytotrans-plantandmaintain.Organizinganozonebioindicatorsiteusingtheseplantswillprovideawaytoobserveozone-inducedfoliarinjuryandtoobserveandmea-sureitsimpactonplantgrowthovertime.Thein-volvementofdifferentgeographicsitesinalong-terminvestigationwillprovidequantitativedatarelativetothesensitivityofthesetwospeciestoambientaircontainingozoneairpollution.Inaddition,theob-servationsbythestudentsinvolvedwithinthisuniqueprojectwilldevelopandincreasetheirawarenessoftheeffectsofozoneairpollutiononsensitiveplantswithinournaturalplantcommunitiesasanimportantpartofourNationalParks.

AstudentchecksCrownbeardplantsinthe“OzoneGarden”atPurchaseKnob,GSMNP,NC.

Dr.HowieNeufeldmeasurestherateofphotosynthesisonozoneinjuredCut-leafconeflowerleavesinGreatSmokyMountainsNationalPark.

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SECTION III: The Ozone Bioindicator GardenBackground Information

OzoneMeasurementsandPlantObservations Participantsineducationalandnon-educationalsettingshavebeentrainedtotakesurfaceozonemeasure-mentswithahand-heldopticalscannerandtogathercloudcoverandtype,humidity,winddirectionandcurrenttemperature.ThesurfaceozoneandmeteorologicaldataaresubmittedtotheGLOBEStudentDataServer(www.globe.gov)atColoradoStateUniversity,FortCollins,CO.Thesurfaceozonedatacanberetrievedinrawformoringraphicvisualizationsenablingthecitizenscientisttoanalyzetheirowndataforpatternsand/orcomparetheirfindingstoothersitescollectingozonedata.OrganizinganOzoneBioindicatorGardencontain-ingozone-sensitiveplantspeciesisanaturalextensiontogatheringsurfaceozonedataandenablestheobservertodevelopanawarenessofhowsurfaceozoneaffectsvegetation. AnOzoneBioindicatorGardenisawayforeducators,students,andcitizenscientistsfromeducationalandnon-educationalsettingstotakepartinanimportantstudytodeterminetheeffectsofozoneairpollutiononplantpopulations.Thestudyofozoneandozone-inducedfoliarinjurywillaidinunderstandingtherelation-shipbetweenfoliarinjury,airquality,andplantgrowth.OzoneBioindicatorGardendataaresubmittedtothe“HandsontheLand”website.http://www.handsontheland.org/environmental-monitoring/ozone-bio-monitoring.html

ItisimportanttoplantCut-leafconeflowerandCommonmilkweedinthenaturalsoilsoftheareawithenoughwalkwayareatopreventdamagingtheplants.Thegardenmustbelocatedawayfromanyareaswherepesticides,herbicides,fertilizers,etcmightbeused.Theobjectiveistomaintainanaturalhabitatforeachoftheozonesensitiveplants.DoNOTuseanyoftheabovechemicalsinyourgardenplotastheymayaffecttheresults.Forexample,somefungicidesarepotentanti-oxidants,andwillkeepplantsfromdevelopinganyozonesymptoms.

ThelayoutofthegardenattheUSForestService’sCradleofForestryinAmericaislinerwithspacebetweenthebedssodatacanbeeasilycollectedwithouttramplingplants.

ThegardenatPurchaseKnobinGreatSmokyMountainsNationalParkisarrangedinrowswith2feetbetweeneachplantand3feetbetweeneachrow.

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No Garden Area? Use Pots Ifyourschoolisinanareaunsuitableforagardenplot,youcanusepots.Itisimportanttouseaslargeapotaspossibletoaccommodatethelargerootsystemoftheplantsandtofinda“breathable”materialliketerracottaorwood.Blackplasticpotswillcausetheplantstodryouttooquicklyinthesunlightsotheyaren’trecommended.Itisnotadvisabletoputmorethanoneplantineachpot.Placethepotsinanareawheretheycangetasmuchsunlightaspossible.Youwillneedtocheckthemdailyforwateringifyouliveinanareathatgetsverywarm.

Planting the Cut-leaf Coneflower Rhizomes Ifyoucan’tputyourplantsintothegroundorpotsimmediately,theywillstoreinarefrigeratorforaboutonemonthifwrappedinmoistpapertowels.Theplantsdobestiftheyareplantedintheearlyfallsotheyhaveachancetoestablishthemselvesbeforeoverwintering.Conefloweroverwinterwitharosetteofbasalleavesthatremainsgreen.Inthespring,theplantwilladdnewbasalleavesandthensendupamainstalk,thisisthepartoftheplantyouwillcollectdataon,notthebasalrosette. Yourrootshavebeencollecteddirectlyoffofplantsfromanexistinggarden.ThisensuresthatalloftheplantshavegeneticmaterialoriginatingwiththegardenatPurchaseKnobinGreatSmokyMountainsNationalPark.Therhizomeshouldstillhaveabitofgreenplantmaterial,whenplanting;thisareashouldbeatthetopandjustbarelycoveredbysoil.Digaholeinthegroundthatisslightlydeeperthenthelengthofanyroothairshangingdownwhenthegreenpartoftheplantisthetop.

Organizing the Plants for Study Oncethedesignofthegardenisestablished,eachplantwillneedtohaveanIDnumberusingCFforCut-leafconeflowerandMWforCommonmilkweed.Alsoconsideradifferentnumberingpatternforeachplantwithinthespecies.Forexample,ifyouhave4Cut-leafconeflowerplants,tagoneasCF01,CF02,CF03andCF04.ThiswillhelppreventconfusingdatafromoneConeflowerplantwithanother,becausethenumbersarenottoosimilar.TheplantIDnumberandleaflabelingpatterndevelopedduringtheinitialsetupofthegardenneedstobethesameeachyearmeasurementsaretaken.

Measuring the Plants AftertheplantIDisestablished,theplantheightneedstoberecordedusingameterstick.• Themeasurementhastoberecordedincentimeters(cm)onthedatasheet.Iftheplantisnotfullygrown,onlymeasureuptothebaseofthetop-mostleaf.Theleavesstanderectwhentheyaren’tfullyopenandifyoumeasuretothetopoftheleaf,theplantmayappeartoshrinkoncetheleafopensfullyanddropsdown.• Countthetotalnumberofleavescomingoffofthemainstemoftheentireplantandrecordobservations(inflower,inbud,nodewithnoleaf,etc).DoNOTcountalloftheleavesontheplant,justthosedirectlycomingoffthemainstem.• Itisimportanttorecognizethatleavesmayfallofftheplantbuttheystillshouldbecountedandincludedindatacollection.Todetermineifaleafismissing,feelalongthestemoftheplantforasmallknob,thisisaleafnodeandindicatesthataleafusedtobethereandwaslost.Workfromthebottomoftheplantto-wardsthetop,runningyourfingersupthestem.• Datacollectionforeachplantwillstartatthebaseoftheplantandwillworkuptowardsthetop.• Identifythenorthsideforeachplant.(Marknorthsomewhereintheplotandwherethebasepointwaswhenmarkingnorthsoeachobserverontheteammayknowforcertainwhichwaytofacewhenrecordingobservations.• Iftheleavesareoppositeeachother,designatetheleavestotheleftofthemiddleoftheplantas“B”andtherightside“A”asyouarefacingNorth.Usingapermanentmarker,putasmalldotonthestemoftheleafclosetowhereitjoinstheplant.Thiswillensure

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thatyoudonotassign“A”toaleafoneweekand“B”tothesameleafthenextweek.Theplantmaytwistasitgrowssothisisanimportantstep.• Iftheleavesalternateonthestem,numbertheleavesinsequencebeginningwithleaf#1-thefirstleafatthebottom,justabovethefloretteofbasalleaves.Ifanyleafhasfallenoff(feelforaleafscarremainonthemainstem),continuetoincludeitinyourcount.

Preparing to Observe Usea10Xmagnifyingglasstocheckleavesandrecordyourobservations.OzoneinjurymaybecomevisibleasearlyaslateMayinareaswithhigherozonelevels,whileotherareasneverexperienceozonesymptoms(andthisisalsoimportantdata).Thefirstsymptomofstipplingontheupperleafsurfacepro-videsthebaselinedateofoccurrenceandinitialdatatobeginassessingfoliarinjury.Datamaybecollectedeverytwoweeks,butitisfinetocollectitweekly. Stopcollectingdataoncetheplantshavebeenthroughtheirfirstfrosteventofthelatesummerorfallseason.Eachleafismeasuredfortheamountof“C”–chlo-rosis(yellowing),“P”–purpling(ozonedamage)and“N”–necrosis(deathofleaftissue).Thepercentageoftheleafthatismissingshouldberecordedinthenotesareaofthedatasheetforeachleaf.

Onthedatasheet,eachleafwillberatedbasedonthefollowingcodes.1-0%2-1-6%3-7-25%4-26-50%5-51-75%6-76-100%Astheexposuretoozoneincreaseandfoliarinjuryincreases,aleafmaybreakoff.Makesuretoassignittheappropriateleaflosscodeusingthefollowingcriteria:7-leafgonewithnopriorsymptoms8-leafgonewithpriorchlorosis(yellowing)only9-leafgonewithpriorpurpling(thebrownishpur-plishdotscalledstippling)only10-leafgonewithpriorpurplingandchlorosis Whenaleaffallsoff,carryitslastdataentryoverintoallofthefollowingdatacollectionentries.Thisistoshowleaflossonagraphorothervisualizationsasopposedtoleafimprovement.Nevermakeassump-tionsaboutwhatyouareseeing,justratewhatyouseeatthatparticularpointintime.Insectswillalsoeatpartofsomeleavesduringtheseason.Ratetheleavesonwhatisvisible,andnoteinthecommentsareaforthatleaftheratingforthepercentageofthetotalleafismissingi.e.2wouldmean1%-6%oftheleafisgone.

Purpling

NecrosisChlorosis

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Training to Assess Foliar Injury An“OzoneBioindicatorGarden”providestheopportunitytomeasuretheamountofozoneinducedinjurytoplantsandtoobservetheimpactofozoneairpollutiononozone-sensitiveplantspeciesovertime.Forthisstudy,weareusingaratingscaledevelopedbytheNationalParkServicetoassessthepercent-ageoffoliarorleafinjurycausedbyozone.AnyoneobservingandrecordingfoliardamageandsubmittingdatatoHandsontheLandwebsitemustparticipateinaformaltrainingtoassessfoliarinjury.Accuracyinassessingfoliarinjuryrequirespracticeandanob-servercanpracticeandrefinehisorherskillsat:http://www.nature.nps.gov/air/edu/O3Training/index.cfm

• Selectthenumberofleafimagesfortraining.Youmaywanttochangethenumberofspecimensto10ratherthan20.• Clickselectifyoumadechanges.• Clickontheimageoftheleafyouwouldliketopracticewith,wesuggestCommonMilkweed.• Selecttheproperpercentrangeandclick“okay”• Anobservermustscoreatleast80%correctwitherrorsbeingnomorethan+or–oneinjuryclassbe-foreproceedingtothefield.• Practiceestimatingfoliainjuryonspeciesforwhichyouwillbecollectingdata.• Youmaywanttopracticeestimatingfoliarinjuryonthewebsiteusingyouridentificationchart.• Haveeachstudentdothisexerciseatleastthreetimesoruntiltheyscore80%.

Registering Your SiteIfyouareaneducatorandarealreadyinvolvedinapartnershipwithapubliclandmanagementagencythatmeetsHOLmembershipcriteria,weencourageyoutoworkwithyouragencycontact(s)tofilloutandsubmittheMembershipApplication.NOTE:Applica-tionsmustbereceivedfromtheagencyprovider.

AgenciesinvolvedintheHandsontheLandnetworkincludetheNationalParkService,NationalFishandWildlifeService,USForestService,BureauofLandManagement,NOAA,EPA,andNRCS.Withineachofthoseagencies,therearemembersites.YoucancontactSusan_Sachs@nps.govforyourapplicationsponsorship.

Entering Data into the Hands on the Land Website

(Note:YouwillneedtobearegisteredmembertoenterdataontheHandsontheLandwebsite)

Gotothedataentrywebsite:http://www.handsontheland.org/environmental-moni-toring/ozone-bio-monitoring.html

1) UnderDatasheet a. Selectyoursite b. ClickonPOST2) Logintoaccessyourdatapage-atthepasswordprompt,enterthefollowing:UserName:youremailaddress Password:selectedbyyouwhenyouinitiallyreg-isteredyoursite.3) Youshouldnowhaveablankdatasheetforyourgarden,checkthegardennameandlocation.Ifyouhavethewrongdatasheet,gobacktothebeginningandmakesureyouhavethepropergardennamese-lectedfromthedropdownmenu4) Writeinthenameofthecollectors,andforgroupsofstudentsteamnamesworkwell.5) Changethedatetothedaydatawerecollected,notthedatetheyareentered.6) Selectthespeciesinitialsfromthedropdownlist BB=Blackberry CB=Crownbeard CF=Cut-leafconeflower CFD=Cut-leafConeflower(var.Digitalis) EB=Elderberry MW=Commonmilkweed WS=Wingstem7) Entertheplantheightincentimeters8) Enterthetotalleafsets(includemissingleaves)9) Enterdataforeachleaf(orleafset).Leaf1isthelowestleafontheplant.Onlyenterdataforthelow-est8leaves.Foralternateleafplants,onlyenterdatacodesinthe“A”column.10)Whenalldatahavebeenchecked,clickthe“In-sert”button.Youwillbeabletoeditandmakecorrec-tionsusingthelinkatthebottomofthepage.

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Ifyouhaveanyproblemswiththissection,contactSusanSachsat(828)[email protected]

Retrieving Data Datacanbeviewedinseveraldifferentways.Youcanviewjustthedata,oryoucandevelopgraphs,animatechangetooneplantovertimeorcomparedatasetsfromoneoruptofourgardens.Onewaytocom-paregardensisbyelevation.Todeterminewhichsitesyouwanttocompare,clickontheGardenMaplinkundertheReports,GraphsandMapssection.Thiswillallowyoutolumpgardensbasedontopography;thusyoucancomparemountaintoptopstovalleysorvalleystoothervalleys.Onceyouhavethesiteloca-tions,youcandothefollowingsearches.

1) Reviewingthedataforjustonegarden:Todothis,clickon“AdvancedSearch”offofthemainozonebio-monitoringpage.Selectthegardenandtheplantspeciesusingthedropdownlist.Fromhere,specifythedaterangeyouwanttoisolate.Areyouwantingtoseeallofthedataorjustoneyearorseason?Hitsearchonceyouaredone. Ifyouwanttoviewdatafromaparticulareleva-tion,youcanenterarangeinthemiddleportionofthesearchpage.Fromhere,enterthedaterangeandhitsearch.

2) ComparingDataforMultipleGardens:Ifyouareinterestedincomparingyourgardenwithuptofourothergardens,click“graph”inthe“compareuptofourdatasetsfromanygarden”intheReports,GraphsandMapssection.a. Firstyoumustselectthegardenstocompareus-ingthedropdownlistthatappearsafteryouhitthe“graph”button.Youmayselectonesiteoruptofour.b. Afteryouselectgardenlocations,hitthe“choosegarden”button.c. Next,chooseaplanttocompare.Inourexample,werandomlychoseleaf#4fromaCut-leafConeflow-erplantfromPurchaseKnob,CradleofForestryandTuscolagardensin2004.Weselectedpurplingasthecharacteristictocompareandthenhit“graph”.d. ThegraphshowedthatonlytheplantatPurchaseKnobshowedsymptomsofpurplingin2004.

3) AnimationofaPlantsFoliarInjuryOverTime:Ifyouareinterestedinseeinganillustrationoftheplantsprogressionoverthegrowingseason,chooseagardenandhit“graph”intheanimationsectionofReports,GraphsandMaps.a. Ifyouhaven’talready,chooseagarden.b. Chooseaplanttoanimatethroughthegrowingseason.c. WerandomlychoseCF0904fromthePurchaseKnobgardenfortheexample.

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Toseethechange,slowlydragthesliderbarunderthegraphicoftheplant.Youshouldseethedateschangeandmightseetheleafcolorschangetoreflectthedatacollected. Theanimationexamplesshowthattheplantshoweddramaticchangesthroughthegrowingsea-son.OnJune16,2004,thefirstdatacollectiondate,onLeaf1theplantshowedaratingof2(1%-6%)forbothchlorosisandpurpling.Leaf4showsaratingof2forchlorosis.BydraggingthesliderbartothethirddatacollectiondateofJune29,wenoticeanincreaseforLeaf1toaratingof4forpurpling(26%-50%)andaratingof2forbothchlorosisandnecrosis.BothLeaf2and3arenowshowingpurplingsymptomsandnecrosisisapparentontheplantLeaf5. BythelastdataentryonJuly26,2004,theplanthaslostthelowest4leavesandthepurplingisvis-ibleontheplantuptoLeaf7witharatingof4(26%-50%).

Looking at Data

Arethedatareasonable? Averageplantinjuryfromozoneairpollutioncanrangefrom0%to100%dependinguponthetimeofyearandthecumulativeozoneexposuresthathaveoccurredthroughoutthespringandsummerseasons.Researchhasshownthatdifferentamountsofozone-inducedfoliarinjuryoccuramongstplantsofthesamespeciesbecauseofthedifferencesinsoil,amountofwater,ozoneexposuresandavailablesunlightontheleafsurfaces.However,therearesomecorrelationsthatusuallyapply:1. Sunlightdrivestheamountofairtakeninforphotosynthesis.Themoreactivetheplantondayswithelevatedozoneairpollution,themorelikelytheozone-sensitiveplantswillshowozone-inducedinjury.

2. Increasedozoneconcentrationsovertimeproducesincreasedplantinjury.Studiesindifferentregionshaveshownthatdifferentelevationsandlevelsofsun-lightinfluencetheseverityofplantinjury.

3. Shadedplantsorevenleaveswillshowlessinjurythanthoseplantsandleaveslocatedinfullsunlightwiththelowerolderleavesshowingmostoftheinjuryduetoseason-longexposures.

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Studentstakingbi-weeklymeasurementsoftheper-centageofplantinjuryshouldobserveageneralincreaseintheamountofdamageovertime.Theyshouldtakeparticularnoteoftheweatherconditions(temperature,dayswithintensesunlight,cloudcoverandtype,winddirection,andhumidity).Thesefac-torsinfluencetheconcentrationsofsurfaceozonepresent.Arethereseveralconsecutivedayswhentheozonelevelishigh?Werethereanydramaticchangesrecordedintheamountsofinjurytotheplants?Onalongertimescale,howdidtheozoneinducedplantinjuryvaryeachmonth?Whatcanbelearnedabouttheeffectsofozoneairpollutionfromtheobservationsmadeoftheincidenceandseverityoffoliarinjurytoplants? Additionalinformationaboutsurfaceozoneandozonedatamaybefoundat:www.epa.gov.Therewillbeadailyairqualitymap.Clickon“Ozone”in

theheadingofthehomepageanditwillprovideback-groundinformationandasitetoclickonforozonelevelswhereyoulive.ThiswilltakeyoutoAIRNOW.Thissiteprovidestoday’sforcastandarchivestore-searchdifferentmonthsandyears.Itdoesnotproviderawdata.YouwillneedtogotoyourstateEnviron-mentalProtectionOfficeorAirQualityofficetogetrawdata.Somestatespostit,butothersdonot.

What do people look for in the data? Onemethodofcollectingdataistoestimatefoliarinjuryonceaweekonthesamedayofeachweek.However,bi-weeklyisalsoacceptable.AsampleofoneseasonofdataforonepairofleavesisrecordedandpresentedinTableI.(Dataispostedonhttp://www.handsontheland.org/environmental-monitoring/ozone-bio-monitoring.html)

Table1:GardenSite:PurchaseKnobPlantID:CB0104Definitions:Chlorosis=yellowing,Purpling=stippling,andnecrosis=deathofleaftissueFoliarAreaInjury:1=0%,2=1%-6%,3=7%-25%,4=26%-50%,5=51%-75%,6=76%-100%

Interpreting Data for Leaf 1A ThisdataisshowingusthatLeaf1Ahadnoimperfectionsofanysortthefirstweek.Duringthesecondweek,therewasaratingof2fornecrosiswhichmeans1%-6%oftheleafhaddeadtissuebuttherewasnootherdamagenoted.Inthethirdweek,7%-25%oftheleafshowedyellowing(chlorosis)and1%-6%oftheleafshowedbothpurplestipplingandnecrosis.Inthenextthreeweeks,thedatastaysthesamewhichindicatestheleaffelloffoftheplantwithpriorpurplingandchlorosisgivingitalosscodeof10.Oncealeaffallsoff,carryoverthepreviousdatasographingdoesnotshowimprovement.

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Table1:GardenSite:PurchaseKnobPlantID:CB0104Definitions:Chlorosis=yellowing,Purpling=stippling,andnecrosis=deathofleaftissueFoliarAreaInjury:1=0%,2=1%-6%,3=7%-25%,4=26%-50%,5=51%-75%,6=76%-100%

Interpreting Data for Leaf 1BThisdatashowsasimilarprogressionas1Aexcepttheleaffelloffwithonlypriorchlorosis,thereforeitgetsaleaflosscodeof8.

Table2:StipplingonLeaves1A,!band2A,2BPlantID:CB0104Definition:Stipplingisthecolorchangethatoccursintheleafduetoozoneinjury.FoliarAreaInjury:1=0%,2=1%-6%,3=7%-25%,4=26%-50%,5=51%-75%,6=76%-100%

Patternsobserved:ThebottomtwopairsofleavesshowednoozoneinjuryduringJune.Beginningdamageof1%-6%begantoshowinJulybutonlyonthetwoleavesontheEastsideoftheplant.TowardstheendJulytheozoneinducedinjuryincreasedtobetween7%-25%.ThesecondleafontheWestsideoftheplantbegantoshowinjurybuttheleaveontheEastsideofthestemoftheplantcontinuetoshowthemostozoneinjury.Theleaflosscodesarenotshowninthistablebuttheyappearasdatacarriedoverinthetableandshowupasastraightlineinthegraph.

Additionalideasfordataanalysis:Dohighozoneepisodesresultinanincreasesinsymptoms?ComparisonsbetweenMWandCFateachsite.Comparisonsbetweenelevationsofthesites.Comparisonsbetweenruralandurbangardensites.TrackingflowerandseedproductionthroughtheyearsMonarchbutterflyfeedingonMWleaves.

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An Example of a Student Research Investigation

Forming a Hypothesis AstudentenrolledwithinaMilanschoolinItalyhasdecidedtofocusontheinterconnectionbetweentheexposuresofozoneairpollutionontheamountofozone-inducedinjurytotheCut-leafconeflower.Shehasdecidedtobeginherresearchprocessbylookingatthevisualizationsofhermeasurementsoftheozoneexposuresandthepatternofozone-inducedplantinjuryduringthemonthsofJulythroughSeptember.Herhypothesisis,theseverityofozone-inducedplantinjuryisdirectlyrelatedtospecificeventsofhighozonelevels.

Collecting and Analyzing Data MonitoringthelevelofozoneinducedplantinjuryatMilanSchoolisanewprotocol,butHandsontheLandpostsseveralmonthsofdatagatheredatvariousgardenlocationsduringthepastyear.ShedecidestorandomlypickaplantfromPurchaseKnob,NCinGreatSmokyMountainsNationalParkandcorrelatetherateofdamagetothe1-hourozoneaverageinppb.Whiledoinganinternetsearch,shediscoversthatthestateofNorthCarolinapostsozonelevelsmeasuredbyprofessionalequipmentthroughoutthestateathttp://daq.state.nc.us/monitor/data/. Shebeginsbyorganizingaspreadsheetofozonemeasurementsfromtheweekpriortoeachdatacol-lectioneventandtheamountsofozone-inducedplantinjuryforPurchaseKnob.ShegeneratesaplotoftheozonemeasurementsusingthecurrentU.S.Environ-mentalProtectionAgencyIndexValuestodeterminehealthyandunhealthyexposurestoozoneairpollu-tion.

Shedeterminesthat65ppbandhigherarerelativelyhighlevelsofozone.Belowisthedatathatthestu-dentexaminedforheranalysis.

WEEK11-hourozoneaveragefortheweekbeforeJune16June09 46ppbJune10 68ppbJune11 60ppbJune12 51ppbJune13 47ppbJune14 51ppbJune15 32ppb

InjuryanimationforJune16

Week1showsozonelevelsbetween32ppband68ppbwith1%-6%damageononlythebottomleaf.

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WEEK21-hourozoneaveragefortheweekbeforeJune22June16 31ppbJune17 34ppbJune18 56ppbJune19 52ppbJune20 59ppbJune21 47ppbJune22 49ppb

InjuryanimationforJune22

Week2showsozonelevelsbetween31ppband59ppbwithLeaf1progressingtoapurplingratingof3(7%-25%)andLeaf2withapurplingratingof2(1%-6%).Theozonelevelswerefairlylowandtheprogressionmovedslowly.

WEEK31-hourozoneaveragefortheweekbeforeJune29June23 41ppbJune24 40ppbJune25 35ppbJune26 52ppbJune27 44ppbJune28 49ppbJune29 52ppb

InjuryanimationforJune29

Week3showsozonelevelsbetween35ppband52ppbwithLeaf1progressingtoapurplingratingof4(26%-50%)andLeaf2withapurplingratingof3andLeaf3developingapurplingratingof2.Theozonelevelswerefairlylowandtheprogressioncontinuestomoveslowly.Thestudentiscuriousaboutthepro-gressionsincethelevelsarewellbelow65ppb.

WEEK41-hourozoneaveragefortheweekbeforeJuly7June30 55ppbJuly1 37ppbJuly2 40ppbJuly3 47ppbJuly4 49ppbJuly5 53ppbJuly6 53ppb

InjuryanimationforJuly7

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Week4showsozonelevelsbetween37ppband55ppbwithLeaf1fallingofftheplant.Leaf2haspro-gressedtoapurplingratingof5(51%-75%)andLeaf3toapurplingratingof3.

WEEK51-hourozoneaveragefortheweekbeforeJuly15July8 57ppbJuly9 57ppbJuly10 60ppbJuly11 57ppbJuly12 51ppbJuly13 58ppbJuly14 59ppb

InjuryanimationforJuly15

Week5showsozonelevelsbetween51ppband60ppb.Comparedtothelastfewweeks,theselevelsaremoreconstantatthehigherendofthehealthyratingsaccordingtoEPA.BothLeaf1andLeaf2havefallenoffandLeaf3hasprogressedfromapurplingratingof3to5.Leaf4isnowshowingpurplingataratingof2,Leaf5and6havedevelopedapurplingratingof3.

WEEK61-hourozoneaveragefortheweekbeforeJuly26July19 61ppbJuly20 73ppbJuly21 67ppbJuly22 71ppbJuly23 66ppbJuly24 60ppbJuly25 52ppb

InjuryanimationforJuly26

Week6showsozonelevelsbetween52ppband73ppb.Whilereviewingherdatafromeachweek,shenoticesthatnoneofthelevelsareparticularlyhigh,only5daysexceed65ppbandnodaysareconsideredunhealthy.Atthesametime,therateofprogressionofsymptomsisfairlydramatic.

Theresultshaveraisedmorequestionsforthestudentthananswers.Sheisnowwonderingaboutthetotalnumberofhourstheplantsreceivedcertainozoneexposures.Thedatashereceivedonlygavethemaximum8-houraveragefortheday.Thestudentdecidesshewantstodomoreresearchontheinternettodetermineifthereisappbstandardforplanthealthsincetheplantsappeartoreacttoozoneinweekswhenozonelevelsareaslowas50ppbatamaximum.Herconclusionisthatsustainedexposureatlowlevelsofozoneexposureseemstobyjustaslikelytopro-ducesymptomsasashort-livedhighexposure.

Further Analysis AnextstepforthestudentfromMilanSchoolmightbetocomparethePurchaseKnobgardenwithoneortwogardensatdifferentelevations.Examinationofresearchshowsthatelevationmayhaveanimpactontherateofsymptoms.Shecouldtrytofindgardenswithsimilarozonelevelsforeachweekandseeifthereisacorrelationbetweenelevation,ozonelevelsandtherateofsymptoms.Anotheranalysiscouldbetolookatthecorrelationbetweentheratesofpurplingandnecrosis.

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ActivitiesBefore Collecting Data

The Leaf Game-Thisactivityallowsstudentstopracticetheestimationskillstheywillneedtocollectdata.

Virtual Ozone Injury Data Collection-SetupontheHandsontheLandwebsitearephotographsfromonefieldseasonatthePurchaseKnobgardeninGreatSmokyMountainsNationalPark.Visithttp://www.handsontheland.org/educator-resources/hol-games/hol-sponsored-games/ozone-inquiry.htmlfortheactivitythatgoesalongwiththeworksheetsonpages32-33ofthisguide.

Collecting Data

Rating Ozone-Induced Foliar Injury on Ozone-sensitive Plants-Thisguidesyouincollectingdata.ChoosetheversionofthedatasheetandOzoneGardenDataCollectionfieldguidethatwillworkbestforyourclass.Also,notethattherearetwodatasheetsforVersionIandII,thesedifferforthevariousplantgrowthforms(alternatevs.oppositeleaves).

Version I-Thisisthesimpliestdatacollectionmethod.Usethisforupperelementaryorremedialmiddle/highschoolgroups.

Version II-Thisistheintermediatedatacollectionmethod.Usethisforadvancedupperelementary,middleschoolorremedialhighschoolgroups.

Version III-Thisisthetrackingdatasheetanditcanbeusedwithadvancedhighschoolorcollegegroups.

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Recordyourresultshere

Trial1=10leaves1.Whichspeciesdidyouuse?

2.Errorsaretobeexpected,butitisimportanttorecordandtrackthecategoriesinwhichyouhavewronganswers.Pleasewritethetotalwronganswersineachcategoryinthespacesbelow.

__0%__1%-4%__5%-12%__13%-25%__26%-50%__51%-75%___76%-100%

Trial2=10leaves1.Whichspeciesdidyouuse?

2.Errorsaretobeexpected,butitisimportanttorecordandtrackthecategoriesinwhichyouhavewronganswers.Pleasewritethetotalwronganswersineachcategoryinthespacesbelow.

__0%__1%-4%__5%-12%__13%-25%__26%-50%___51%-75%___76%-100%

Trial3=10leaves1.Whichspeciesdidyouuse?

2.Errorsaretobeexpected,butitisimportanttorecordandtrackthecategoriesinwhichyouhavewronganswers.Pleasewritethetotalwronganswersineachcategoryinthespacesbelow.

__0%__1%-4%__5%-12%__13%-25%__26%-50%__51%-75%___76%-100%

Trial4=10leaves1.Whichspeciesdidyouuse?

2.Errorsaretobeexpected,butitisimportanttorecordandtrackthecategoriesinwhichyouhavewronganswers.Pleasewritethetotalwronganswersineachcategoryinthespacesbelow.

__0%__1%-4%__5%-12%__13%-25%__26%-50%__51%-75%___76%-100%

Didyougetanybetterwitheachtrial?Inordertocollectdataintheozonegarden,youmustgetatleast80%correctononeofthe4trialsandbenomorethanonecategoryoffonincorrectanswers.

TheLeafGameNavigatetotheFoliarInjuryAssessmentModulewebsite:http://www.nature.nps.gov/air/edu/O3Training/index.cfm• Changethenumberofleafimagesfortrainingto10• Click“select”• StarttrainingonCommonMilkweed

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O3 Skills Center 1Parks as Classrooms - Great Smoky Mountains National Park

Name ______________________________

O3 Skills Center Student WorksheetGo to the O3 Inquiry website: http://www.handsontheland.org/ozone-inquiry

Answer the following questions in a few phrases or in a sentence. 1. What does chlorosis look like?

2. What does chlorosis mean for the plant?

3. What does necrosis look like?

4. What does necrosis mean for the plant?

5. What does purpling/stippling look like?

6. What does purpling/stippling mean for the plant?

7. What is happening inside of the leaf?

8. How do we know that the symptoms are from ozone?

9. Choose one Plant in the drop down menu. Circle the one chosen: Plant 1, Plant 2, Plant 3, Plant 4

10. Choose one of the following Dates in the drop down menu. Circle which one you chose: June 29, July 6, July 13, July 20, July 27, August 4

11. Complete the following table using the following foliar area injury codes: 0 = leaf missing 1 = 0% 2 = 1%-6% 3 = 7%-25% 4 = 26%-50% 5 = 51%-75% 6 = 76%-100%

Chlorosis Purpling NecrosisLeaf A Leaf B Leaf A Leaf B Leaf A Leaf B

Set 8Set 7Set 6Set 5Set 4Set 3Set 2

Set 1

12. Insert the above data into the O3 Skills Center website and click the “Check” button. What is your score? ________

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12. Insert the above data into the O3 Skills Center website and click the “Check” button. What is your score? ________

13. When was the first observed date the selected plant flowered (click at the top of the plant for each week to determine if the plant has flowered)? Circle which date the selected plant flowered: July 27, August 4, August 11, August 31, September 7, September 14

14. Choose another Date (use a date listed below) in the drop down menu using the same selected Plant as before. Circle which one you chose: August 11, August 31, September 7, September 14, September 20, September 28

15. Complete the following table using the following foliar area injury codes: 0 = leaf missing 1 = 0% 2 = 1%-6% 3 = 7%-25% 4 = 26%-50% 5 = 51%-75% 6 = 76%-100%

Chlorosis Purpling NecrosisLeaf A Leaf B Leaf A Leaf B Leaf A Leaf B

Set 8Set 7Set 6Set 5Set 4Set 3Set 2

Set 1

16. Insert the above data into the O3 Skills Center website and click the “Check” button. What is your score? ________

17. Describe three differences you observed between the two dates selected. 1. 2. 3.

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Lesson Plan: Rating Ozone-Induced Foliar Injury on Ozone-sensitive PlantsPurposeTomeasurethepercentageofozone-inducedinjurytoplants.

Overview• Determineanaveragepercentareaoftheozone-in-ducedinjuryoneachplantleaf.• UsetheNationalParkService’sratingscaletoquantifytheareaofleafaffected.Theratingclassesof1-6representapercentagerangeoffoliarinjury(1=0%,2=1-6%,3=7-25%,4=26-50%,5=51-75%and6=76-100%)andwillbeidentifiedusingafieldguidefortheassessment.

Educational OutcomesParticipantswilllearntoidentifythepercentofozone-inducedinjuryandchangesoccurringtotheplantovertimeasozonepollutionexposureschange.

Science, Geography, Mathematics ConceptsInquiry• Askaquestionaboutplantinjuryandeventsintheenvironment• Planandconductaninvestigation• Usedatatoconstructreasonableexplanations• MaintainajournalPersonandSocialPerspectives• Changesintheenvironmentcanbenaturalorinflu-encedbyhumans• MaterialfromhumansocietiesaffectbothphysicalandchemicalcyclesofearthEarthandSpace• Theatmosphereismadeupofdifferentgases• MaterialsfromhumansocietiesaffectthechemicalcyclesoftheEarthLifeScience• Cellscarryonmanyfunctionsneededtosustainlife.• Demiseisabreakdowninthestructureandfunc-tionsofanorganismGeography• Humanactivitycanmodifythephysicalenviron-ment.Mathematics• Understandandapplyratios,proportions,andper-centsinawidevarietyofsituations• Formulateandsolveproblemsthatinvolvecollect-ingandanalyzingdata• Construct,read,andinterpretdisplaysofdata• Communicateresults

Observation SupportOzoneinducedfoliarinjuryiscommonlymeasuredusingtheNationalParkService’sratingscale.Thegraphicchartidentifiesthevaryingpercentageofozoneinjurytoaplantusingtheratingsof:0%,1-6%,7-25%,26-50%,51-75%and76-100%.Thegreatertheamountofozoneairpollutionpresent,themorefo-liarinjurythatwilllikelybeobservedonozone-sensi-tiveplants.Theplantisobserveddailyusinga10Xmagnifyingglassuntilthebeginningofstipplingisseenonaleaf.Theplantdataarethenrecorded.ThefirstdatacollectingeventshouldoccurnolaterthanJune15eachyear,subsequentobservationsshouldbetakenonedayeachweekoreverytwoweeks,tryingtomaintainaregularschedule.Remember,noinjuryisrelevantdata.

Theamountoffoliar(leaf)injuryislikelytoincreaseshortlyafterdayswithhighlevelsofozoneairpollution.Thiswouldbeanopportunetimeforadiurnalstudyofsurfaceozoneandcorrelatingthatdatawiththelevelofinjuryobservedontheleafoftheplant.Iftakinghourlyozonemeasurementsisnotpossible,thehourlymeasurementsmaybegottenfromthelocalEnvironmentalProtectionAgencyorlocalairquality-monitoringdevice.Aninternetsearchwillrevealifyourstatepostsairqualitydata.

Measurement LogisticsTheneedtoassesstheplantdailyforozoneinjuryuntilthefirstsymptoms,mayposealogisticschal-lenge.Oneapproachmightbetousethemagnifyingglasstocheckthelowestleavesoftheplantduringarecessorlunchbreak,orimmediatelyafterschoolisout.Identifyingthedayeachoftheplantsshowsymp-tomsoffoliarinjuryandratingtheamountofinjuryprovidesabaselineofdataformonitoringthepatternoffoliarinjuryovertime.

Afterestablishingabaselineofdata,theplantisassessedonceaweekpreferably,oreverytwoweeksonthesameday.Establishaschedulesoeveryoneinvolvedknowswhatisexpectedandwhentodoit.

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Supporting Measurements Thepercentageofinjurytoaplantdependsupontheleveloramountofozonepresentandtheamountofsunlight.Strongsunlightsupportsphotosynthesis,producingfoodtheplantneeds.Thisrequiresmoreintakeofairbytheplant,andincreasedintakeofaircontaininghigherlevelsofozonedamagetheplant.Hotsunnydays,hightemperatures,winddirection,humidity,andcloudcoverallaffecttheconcentrationofozoneproduced.Levelsofozonecanbemeasuredwithahand-heldopticalscannerusingtheSurfaceOzoneProtocolfoundintheGLOBE(GlobalLearn-ingandObservationtoBenefittheEnvironment)Pro-gram(www.globe.gov).ParticipatinginthetrainingfortheozoneprotocolenablesyoutosubmityourdatatotheGLOBEStudentDataServer.Thedatacanberetrievedinrawformorthroughvisualizationsmakingitanaturalextensiontothestudyofplantinjury.

Participant PreparationAnyoneassessingfoliar(plant)injuryandsubmit-tingdataHandsontheLadwebsitemustbetrained.Itisimportanttotheaccuracyofthemeasurementsthatparticipantsareableto:1.Workincooperativegroupsof2-4tosetupthesiteandgather,analyze,anddiscussresults.2.Organizeallmaterialsneededtoassessplantinjury.3.Followaschedulefortakingmeasurements.4.Identifyandrecord,date,time,gardenlocation,plantIDnumber,numberofleavesontheentireplantandotherobservationsrequiredonthedatasheet.5. EstimatethepercentageoffoliarinjuryusingthegraphicallydesignedchartsummarizingtheNationalParkService’sratingscale.6.Practiceestimatingfoliarinjuryusingthewebsite:http://www.nature.nps.gov/air/edu/O3Training/index.cfm7. Recorddataaccuratelyandcompletelyforsubmit-tingtotheHandsontheLandwebsite.

Helpful Hints» Designateanareatokeeptheclipboard,datasheetandallmaterialsforgatheringobservations.» Checkthewrittenrecordsfromtimetotimeforcompletenessandaccuracy.

Questions for Further InvestigationHowistheamountoffoliarinjuryrelatedtootheratmosphericphenomena?Whichone?How?

Whatvariabilityoffoliarinjurydidyouobserveoveraperiodoftime?Daily?Weekly?Seasonally?

Howcouldyouuseyourdataonplantinjurytochar-acterizeanychangesintheozoneexposures…weekly?,seasonally?,betweenyears?

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Site Definition Choosingthelocationforyour“OzoneBioindica-torGarden”andarrangingtheozonesensitiveplantsareimportanttoyoursuccessfulimplementationofobservingplantsforozoneinjury.Assessmentofplantinjurybeginswithusinga10Xmagnifyingglasstoobserveeachplantatthebeginningoftheozoneseason(MayorJune)fordeterminingtheinitialsymp-tomsofinjury.Observersneedtobeabletogetclosetoeachplantwithoutwalkingonordamaginganotherplant.Theidealgardenmaybecircularorrectangularwith4plantseachofthesamespeciesplanted18inchesapart.EachplantistaggedwithanumberusingCFforconeflowerandMWformilkweed.Placeametalrodwitharedflagwiththeindividualplant’snumberontheflag,atthebaseofeachplant.Thiswillalsobeamarkertoidentifythenewplantforthenextyear.OnceeachplanthasbeengivenanIDtag,identifyNorth.UsingNorthasthefocalpoint,eachleafoneachplantwillneedtobeidentified.Startatthebot-tomoftheplanttonumbertheleaves.Ifaleafhasfallenoffoftheplant,itshouldstillbecountedandlabeled.Thelastdataforthatleafwillcarryoverfromweektoweekthroughoutthedatacollectionperiod.

Plantwithparallel/oppositeleaves• Leavesaredirectlyacrossfromeachother,theleavestotherightofthestem(facingNorth)willbelabeled1A,2A,A3,4A,5A,6A,7A,and8A.• Theleavesonthesameplanttotheleftofthestem(youfacingNorth)willbelabeled1B,2B,3B,4B,5B,6B,7B,8B.

Anopenareaofatleast15-20metersonasidesquareisnecessarytoallowgoodairmovementandnormalprecipitationwithout“rainshadows”,andofgreatestimportance,theplantmustreceivedirectsunlight.Usethenaturalsoilintheareaforplantingeachspeciesintheirowngarden.Thegarden’sloca-tionmustbeawayfromareaswherepesticidesandfertilizersareused.Itisimportantthatnopesticidesorfertilizersareusedinthe“OzoneBioindicatorGar-den”.Inselectingyoursite,somecompromisemaybenecessarybetweentheidealsiteforscientificobserva-tionsandthelogisticalconstraintsoftheareaanditssurroundingsofthesiteavailabletouse.Toensurethevalueofyourdatadocumentthenatureofyourbioin-dicatorsiteanditssurroundings. Thespreadingoftheplantsiscontrolledbydigginguprootsattheendoftheseason,andtrimmingbackanyexcessgrowth.Youmaywanttoletasecondplantgrownexttoeachoftheoriginalplantstodoubleeachspeciesfordatagathering.Ifnot,thentrimbackeachoriginalplant.Besuretoplaceasmallflaginthelocationofthegrowthofeachplantandhavetheplant’sIDnumberontheflag.Also,designateandbeconsistentyeartoyearwithwhichsideoftheplantis“A”or“B”.MakeamapofthelayoutofyoursiteandmarkN,S,E,andWandthelocationofeachlabeledplant.

Plantwithalternatingleaves• Leavesalternateonthestem.Onlythe8bottomleavesarenumbered.• Ifaleafhasfallenoffnearthebot-tomoftheplant,itiscountedasoneofthe8bottomleaves.• Startingfromthebottom,feelalongthestemtomakesureyouarecount-ingleafscarsaswell.Thefirstleafaboveanybasalleavesis1.Continuenumberingleavesupthemainstemuntil8.

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Site Definition SheetOzone Biomonitoring Garden

SchoolName:________________________ ClassorGroupName_____________________________

Name(s)ofstudent(s)fillinginSiteDefinitionSheet:______________________________________________

Date:________________ Checkone:ConeflowerSite MilkweedSite

SiteName:(Giveyoursiteauniquename)_______________________________________________________

Location:Latitude:_______________NorS Longitude___________________EorW

Elevation:__________meters

SourceofLocation(Checkone)GPS Other(pleasespecify)______________

Describesite:(Includetrees,buildings,etc.thatarenearyoursite).

SendapictureofyoursiteandorientationofgardenlayouttoHandsontheLand.

Drawamodelofyourgardenlayout,labelNorth,South,EastandWest,andplacemarkersingardentoidentifyN,S,E.andW.Numbereachplantandplaceamarkeratthebaseofeachplant.Youwillusethesamenumberfortheplantthatgrowsinthesameareathefollowingyear.

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Field InstructionRating Ozone-Induced Foliar Injury

Materials• Meterstick• Tagswithstring,andblackpermanentmarkers• Sitelocationflagforeachplantwiththeplantnum-berontheflag• Metricmeasuringtape• %ozoneleafinjuryidentificationchart• ClipboardandPencil• Smallhandmirrortoplaceundertheleaftoseeifdamagegoesthroughleaf• Small10XhandlensNOTE:ThefirstevaluationsshouldoccuratleastbyJune15.Thefinalevaluationshouldbebeforethetypicalfirsthardfrostforyourarea.

ProceduresforPlotEvaluation1.Begincollectingdataonplantsoncetheytallenoughtohaveatleastthreesetsofleavesgrowingfromthemainstem.Thisdatewillvaryaccordingtolocale.

2.Measuretheindividualheightsoftheplantsfrombasetoapex.usingameterstickandrecordheightincentimeters,athinpieceofwoodshouldbecarriedtothebaseofeachplanttopreventthemeterstickfromdepressingintotheground.

3.Whenmeasuringtheheightoftheplant,measureuptothebottomofthehighest,mostopenleaf.Oncetheplantisinbudorflower,holdtheplantstraight,followthemainstem(thethickestbranch)andmea-suretothetopofthetallestbloomonthemainstem.

4.Whencollectingdataontheindividualleaves,startatthebaseoftheplant.

5.Countthetotalnumberofleavesontheplantthatcomedirectlyoffofthemainstem.Donotcountapicalleavesthatarenotfullyexpanded.(Ingeneral,evaluateonlyleavesthatareatleast65%expanded).Donotcountleavesthatareonsub-branchestothemainstem.Docountleafscarswhereleaveshavefallenoff(runyourhandupthestemtofeelforleafscars).

6.Estimatethepercentageoftotalleafareaofeachleafthathasozone-inducedinjury.Ratingclassesare1=0%,2=1%-6%,3=7%-25%,4=26%-50%,5=51%-75%,6=76-100%.TheplantsareratedonC=clorosis(yellowing),P=purpling(stippling),N=necrosis(death).Necrosisisvisibleonbothsidesofaleafunlikepurplingwhichonlyappearsontheupperleafsurface.

7.Asozoneinjuryprogresses,purplingwillturnintonecrosis.Soitdoesn’tappearthattheplanthasimproved,itisimportanttocarryoverthepreviousweek’sdataforpurpling(e.g.ifaleaf’snecrosisratinggoesuptoa5,thepurplingmaynolongerbeappar-ent.Ithasn’tgottenbetter,ithasjustprogressedtoapointwhereithaschangedfrominjurytodeadtissue.Carryoverthelastdataentryforpurpling).

8.Ideally,datashouldbecollectedonceaweek.Ifthatisnotpossible,tryforatleastoncepermonth.

9.Asthesummerseasoncomestoaclose,recordthenumberofflowersandseedpodsperplant.

10.Forplantswithoppositeleaves,eachfacingpairofleavesisoneset.Foralternateleafplants,suchastheconeflower,countonlytheleavescomingoffofthemainstem.

11.Usecautionwhencheckingthebackoftheleaf,especiallyifyounoticeanysymptoms.Theseleaveswillbecomefragileandwillbreakoffeasily.Getdowncloseandgentlyturntheleafovertomakeyourobservationoruseahandmirrotoreflecttheunder-side.

12.Ifaleafbreaksoff,makesuretoassignittheap-propriateleaflosscode.7-leafgonewithnopriorsymptoms8-leafgonewithpriorchlorosis(yellowing)only9-leafgonewithpriorpurplingonly(ozone)10-leafgonewithpriorpurplingstipplingandchlorosis

13.Whenaleaffallsoff,carryitslastdataentryoverintoallofthefollowingdatacollectionentries.Thisistoshowleaflossonagraphasopposedtoleafim-provement.

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14.Nevermakeassumptionsaboutwhatyouaresee-ing,justratewhatyouseeatthatpointintime.(Whatofinsectfeedingorsomeothersymptomthatisuncer-tain,contactSusanSachsat:[email protected]?Includedigitalphotosifpossible?

15.Duringtheseason,insectswilleatpartofsomeleaves.Ratetheleafsurfaceareathatisvisible,butthenalsonoteinthecommentsareaforthatleafthepercentageofthetotalleafareamissing.

16.Studentsneedtobeawareofmimickingsymptomsandotherpestproblemsthatlooklikeozoneairpollu-tioninjurytosensitiveplants.Somestrategiesare: a.Lookatolderleavesastheyshouldhavethemostinjury.Ifyoungerleaveshavethemostinjury,thenitisnotozonedamagethatyouareobserving. b.Thepurplestipplingshouldbeonthesurfaceoftheleafonlyandnotontheundersideoftheleaf. c.Thepurplestippling(orsmallpurpledots)donotenteranyoftheveins. d.Beawareofmimickingsymptoms,andotherpestproblemsthatlooklikeozoneinjury.

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Injury level 10%

Lower end of Injury level 2 (1-6%)

Lower end of injury level 3 (7-25%)

Upper end of Injury level 2 (1-6%)

Upper end of injury level 3 (7-25%)

Injury

Injury

41

Lower end of injury level 4 (26-50%)

Lower end of injury level 5 (51-75%)

Injury level 676-100%

Upper end of injury level 4 (26-50%)

Lower end of injury level 5 (51-75%)

Must notice the small green Areas that make this leaf a

high Level 5 as opposed to a 6

Complete foliar injury

42

Leaf Field GuideEstimating Foliar Injury-Common Milkweed

Usethefollowingcharttoguidetheestimationoffoliardamagetoeachleaf.Remember,thestippling(alsocalledpurpling)isonlyonthetopsideoftheleafandthediscolorationdoesnotoccuronanyoftheleafveins.NationalParkServiceCategorizationMethod

43

Garden Location: Plant ID #:

Plant species: Plant height (cm):

Total # of leaves on the entire plant:

Observations (inflower,inbud...)

Ozone Bio-monitoring Garden Data Sheet - Alternate Leaves

Group Name: Date:

Ratethe%ofozonesymptomscoveringtheentireleafP=purpling(reddish-purpleinjuryspotscausedbyozone)

1

2

3

4

5

6

7

8

TOP OF PLANT

BOTTOM OF PLANT

�0%1-6%7-25%26-50%51-75%76-100

leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

0%1-6%7-25%26-50%51-75%76-100leafpresent missing

P

Observations

AlternateLeafDataSheet-VersionI

44

Ozone B

io-monitoring G

arden Data Sheet

- Opposite L

eavesSchool:___________________________

Date:___________

Plant species: ____________Plant ID

#: ________________

Plant height (cm): _____________ Total # of leaf sets on the entire plant: __________ O

bservations (inflower,inbud...)___________

Current 1 hour ozone:___________

______________________________________

4A4B

5A5B

6A6B

7A7B

8A8B�

TOPOFPLANT

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

1A2A

3A

1B2B

3B

BOTTOMOFPLANT

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

leafgoneP

10%

21-6%

37-25%

426-50%

551-75%

675-100

notes

Oppositeleafdatasheet-VersionI

45

Garden Location: Plant ID #:

Plant species: Plant height (cm):

Total # of leaves on the entire plant:

Observations (inflower,inbud...)

Ozone Bio-monitoring Garden Data Sheet - Alternate Leaves

Group Name: Date:

Ratethe%ofozonesymptomscoveringtheentireleafP=purpling(injurydots),C=clorosis(yellowing),N=necropsy(death)

1

2

3

4

5

6

7

8

leafpresent missing0%1-6%7-25%26-50%51-75%76-100

CPNObservations

TOP OF PLANT

leafpresent missing0%1-6%7-25%26-50%51-75%76-100

CPNObservations

leafpresent missing

leafpresent missing

leafpresent missing

leafpresent missing

leafpresent missing

leafpresent missing

0%1-6%7-25%26-50%51-75%76-100

0%1-6%7-25%26-50%51-75%76-100

0%1-6%7-25%26-50%51-75%76-100

0%1-6%7-25%26-50%51-75%76-100

0%1-6%7-25%26-50%51-75%76-100

0%1-6%7-25%26-50%51-75%76-100

CPNObservations

CPNObservations

CPNObservations

CPNObservations

CPNObservations

CPNObservations

BOTTOM OF PLANT

�

AlternateLeafDataSheet-VersionII

46

OppositeLeafDataSheet-VersionII Ozone Biomonitoring Data Sheet - Opposite LeavesCollector Names: _____________________________________________________ Date:_________________

Garden Location: ______________________________

Plant Species: _____________________________________ Plant ID#:_______________________

Plant Height (cm): _________________________________ Total # of leaf pairs: ______________

Observations (in flower, in bud...) _____________________________________________________________

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

8A

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

Leaf present Leaf missing

0% 1-6% 7-25% 26-50% 51-75% 76-100%

P

C

N

7A

8B

6A

5A

4A

3A

2A

1A

7B

6B

5B

4B

3B

2B

1B

Bottom of Plant P= purpling (injury dots) C=chlorosis (yellowing) N=necropsy (death)

47

Ozone B

io-monitoring G

arden Data Sheet

DataC

ollectors:_______________________________________________________________

GardenLocation:_________________________________

PlantSpecies:_______________________PlantID

:__________________________

Foliar Area Injury Survey

1=0%,2=1%

-6%,3=7%

-25%,4=26%

-50%,5=51%

-75%, 6=76%

-100%(N

PSscale)L

eaf Loss codes

7=leaffelloffwithnoratinghigherthan1,

8=leafhadpriorCbutnoP,

9=leafhadpriorPbutnoC,

10=leafhadpriorP&C

Leaf

Flwr

Leaf

set(y/n)

pairA

24

1-

14

1-

13

1-

13

2-

12

1-

21

2-

--

--

--

--

7/1/200331cm

12n

B1

41-

13

1-

13

1-

13

1-

12

1-

11

17-

--

--

--

-

Date

Ht.

lf setFlw

rAB

Date

Ht.

lf setFlw

rAB

Date

Ht.

lf setFlw

rAB

Date

Ht.

lf setFlw

rAB

Leaf set=totalnum

berofleavesontheentireplantL

eaf Pairisforthoseplantsthathaveleavesoppositeoneanotheri.e.milkw

eedC=clorosis(yellow

ing)P=purpling(stippling)

N=necropsy(deathofleaftissue)L=leaflosscode

•Ifyouentera6fornecropsyoranyvalueunder“L”,carryoverthepreviousweeksratingforC

&P

•Justratewhatyousee,don’tm

akeanyassumptionsastow

hyyouareseeingit.

NO

TE

S

examplecollection __collection __collection __collection __ Date

Ht.N

OT

ES

NO

TE

S

NO

TE

S

NO

TE

S

Leaf 8

C P N

L

3missing

leafbroken2m

issing

Leaf 6

C P N

LL

eaf 7L

eaf 3C

P N L

C P N

LL

eaf 4C

P N L

Leaf 5

C P N

L L

eaf 1(low

est)C

P N L

Leaf 2

C P N

L

DataSheet-VersionIII

48

49

50

51

52

53

54

TheseWebpagesshouldprovehelpful,andwillpro-videmanyotherlinks.

1.www.epa.gov

USEnvironmentalProtectionAgencyhomepage:Lotsofinformationregardingairquality(standards,emis-sions,effects,etc.),climatechange,ecosystemeffects.

2.www.noaa.gov

NationalOceanic&AtmosphericAdministrationhomepage:Goodinformationregardingairquality(mainlyatmosphericchemistry&meteorology),cli-matechange.

3.www.ncar.ucar.edu

NationalCenterforAtmosphericResearchhomepage:Lotsofgoodinformationrelatedtoglobalclimatechange,carbonbudgets,&atmosphericchemistry.De-scriptionofalarge-scaleglobalclimatechangemodel.

4.www.ncl.ac.uk/gane

Webpagerelatedtoamajorprogramdealingwithnitrogendeposition&effects.Followthelinksfordetailedinformation.

5.www.ncl.ac.uk/airweb

Webpagerelatedtoadescriptionofbiologicaleffectsforozone,fluoride,andsulfurdioxide.Followthelinksfordetailedinformation.

6.www.ipcc.chIntergovernmentalPanelonClimateChangehomep-age.Somegoodinformationrelatedtoglobalclimatechange.

7.www2.nature.nps.gov/air/ NationalParkService’swebsiteonairqualityinparks.Includesissues,dataandwebcameras.

8.nadp.sws.uiuc.edu.

AdescriptionoftheNationalAtmosphericDeposi-tionProgam/NationalTrendsNetwork(NADP/NTN)nationalnetworkformonitoringacidicdeposition.

9.www.asl-associates.com.

Webpagededicatedtoinformationaboutairpollution,primarilyozone,particulatesandsulfurdioxide.

10.www.epa.gov/airnow.

Realtimeimagedemonstrationofozonebuildupovertimeandcategoriesofseverity.

11.www.ace.mmu.ac.uk.

GototheResourcesIndex.Therearemanydifferentfactsheetsandteachingresourcesthatwillbeofhelptoyou.Isuggestyoulookatthissite.

12.www.ozone.wsl.ch

Webpageshowingvisibleozoneinjuryofseveralsen-sitiveplantsandhowtousethisinformationinassess-ment.

13.www.fia.fs.fed.us&www.fiaozone.net/index.html

InterestinginformationconcerningtheUSDAForestService’sForestHealthMonitoringProgram.

14.http://reports.eea.eu.int

EuropeanEnvironmentAgencyWebsite.

15.www.heinzcenter.org/ecosystems/index.htm

AreportonthestateoftheNation’secosystems.Goodinformationregardingindicatorsofecosystemchange.

16.mona.psu.edu

Alinktoafoliarinjuryassessmentmodule.Atrainingtoolusedforindividualsintheassessmentofairpollu-

Some Useful WebpagesProvidedbyDr.ArtChappelka

55

tioninjurytovegetation.

17.www2.nature.nps.gov/air/pubs/index.htm.

AlinktovariouspublicationsrelatedtoairqualitybytheNPS.

18.airlichen.nacse.org/.

Thisisagoodwebsiteforinformationonlichensasbioindicatorsofairpollution.

19.toxmap.nlm.nih.gov/toxmap/main/index.jsp.

Thiswebsiteprovidessomeinformationandmapsregardingenvironmentalhealthandtoxicology.

20.reports.eea.eu.int/EMEPCORINAIR4/en.

Linkstovariousinventorymodels,etc.inEurope.

56

REFERENCES

Horsefall,J.G.,Barreatt,R.W.1945.AnImprovedGradingsystemforMeasuringPlantDisease. Phytopathology35,655.

Duchelle,S.F.,Skelly,J.M.andKress,L.W.1980.Theimpactofphotochemicaloxidantairpollutiononbio massdevelopmentofnativevegetationandsymptomexpressionofAesclepiasspp.Phytopathol.70:689.

Duchelle,S.F.andSkelly,J.M.1981.TheresponseofAsclepiassyriacatooxidantairpollutioninthe ShenandoahNationalParkofVirginia.PlantDisease65:661-663.

Skelly,J.M.,Davis,D.D.,Merrill,W.,Cameron,E.A.,Brown,H.D.,Drummond,D.B.,andDochinger,L. S.(eds.)1987.DiagnosingInjurytoEasternForestTrees.USDA-For.Ser.,Veg.SurveyRes.Coop.and PennStateUniv.,UniversityPark,PA.122p.

Hildebrand,E.S.,Skelly,J.M.andFredericksen,T.S.1996.Foliarresponseofozonesensitivehardwoodtree speciesfrom1991-1993intheShenandoahNationalPark,VA.Can.J.For.Res.26:658-669.

Neufeld,H.S.,Renfro,H.S.,Hacker,W.D.,andSilsbee,D.1992.OzoneinGreatSmokyMountainsNational Park:DynamicsandEffectsonPlants.In“TroposphericOzoneandtheEnvironmentII”,editedbyR.L. Berglund,Air&WasteManagementAssoc.,pp.594-617

Skelly,J.M.1998.AbriefassessmentofforesthealthinnortheasternUnitedStatesandsoutheasternCanada. NortheasternNaturalist5(2)137-142.

Skelly,J.M.2000.Troposphericozoneanditsimportancetoforestsandnaturalplantcommunitiesofthenorth easternUnitedStates.NortheasternNaturalist(3)221-236.

Innes,J.L.,Skelly,J.M.andSchaub,M.2001.Aguidetotheidentificationofozone-inducedfoliarinjuryon broadleavedtree,shrubandherbspecies.PaulHauptPublishing,Bern,Switzerland.ISBNNo.3-258- 06384-2.136pp.

Chappelka,A.H.,Neufeld,H.S.,Davison,A.W.,Somers,G.L.,andRenfro,J.R.2002.Ozoneinjuryoncutleaf coneflower(Rudbeckialaciniata)andcrown-beard(Verbesinaoccidentalis)inGreatSmokeyMountains NationalPark.EnvironmentalPollution125(2003)pp.53-59.

Orendovici.T.,Skelly,J.M.,Ferdinand,J.A.,Savage,J.E.,Sanz,M-J.andSmith,G.C.2003.Responseof nativeplantsofnortheasternUnitedStatesandsouthernSpaintoozoneexposures:Determiningexposure/re sponserelationships.EnvironmentalPollution125:31-40

Yuska,D.E.,Skelly,J.M.,Ferdinand,J.A.,Stevenson,R.E.,Savage,J.E.,Mulik,J.D.andHines,A.2003. Useofbioindicatorsandpassivesamplingdevicestoevaluateambientozoneconcentrationsinnorthcentral Pennsylvania.EnvironmentalPollution125:71-80.

Skelly,J.M.2003.Nativeplantsasbioindicatorsofairpollutants:contributedpaperstoasymposiumheldin conjunctionwiththe34thairpollutionworkshop.EnvironmentalPollution125:1-2.