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Food
AgencySummaryMemorandumRe:ConsultationwithCalgene,Inc.,ConcerningFLAVR
SAVRTomatoes
FoodMasterFiles&FoodAdditivePetitionsMainPage14
Date:May17,1994
From:ConsumerSafetyOfficer,BiotechnologyPolicyBranch,HFS206
Subject:SummaryofconsultationwithCalgene,Inc.,concerningFLAVRSAVRtomatoes
To:ActingDirector,OfficeofPremarketApproval,HFS200
InaletterdatedAugust12,1991,Calgene,Inc.,requestedanadvisoryopinionunder21CFR10.85
concerningwhetherFLAVRSAVRtomatoesarefoodand,therefore,subjecttothesameregulationas
othertomatovarieties.InanoticepublishedintheFederalRegisterofMay29,1992(57FR22984),FDA
issuedapolicystatement(the1992policystatement)clarifyingtheagency'sinterpretationoftheFederal
Food,Drug,andCosmeticAct(theact)withrespecttofoodsderivedfromnewplantvarieties.Inlightof
thepublicationofthe1992policystatement,webelievethatanopinionconcerningthestatusofa
particularproductsuchasFLAVRSAVRtomatoesisbestaddressedthroughaconsultationwiththe
agencyconsistentwiththeprinciplesoutlinedinthatstatement.Therefore,werecommendthattheOffice
ofPremarketApproval(OPA)respondtoCalgene'srequestasaconsultationinaccordancewiththe1992
policystatementratherthanasanadvisoryopinionunder21CFR10.85.Thismemorandumsummarizes
thatconsultation.

INTRODUCTION

BackgroundInformation
ApproachtotheEvaluationofFLAVRSAVRTomatoes
OverviewofthisMemorandum

FOODSAFETYASSESSMENTCONDUCTEDBYCALGENE

SummaryofDataSubmittedbyCalgene
BackgroundInformationonTomatoes
TheIntendedEffectoftheAntisensePGGene
BackgroundInformationontheGeneTransferSystem
SourcesofDNAIntroducedintoFLAVRSAVRTomatoes
GeneticStability
NutrientsinFLAVRSAVRTomatoes
GlycoalkaloidToxicantsinFLAVRSAVRTomatoes

NewProteinsIntroducedintoFLAVRSAVRTomatoes
AnimalGavageStudies
Introduction
Results
GastricErosions
Conclusions

STATUSOFFLAVRSAVRTOMATOES

ENVIRONMENTALCONSIDERATIONS

LABELING

StatutoryandRegulatoryRequirementsforLabeling
LabelingFoodsDerivedfromNewPlantVarieties
RequirementsforLabelingFLAVRSAVRTomatoes

EVALUATIONOFCOMMENTS

Overview
RegulatoryIssuesRaisedintheComments
Premarketnotification
Premarketapproval
FDAregulationsconcerningadvisoryopinions
ThestatusofnewsubstancesintroducedintoFLAVRSAVRtomatoes
ThescopeofFDA'sresponse
Labeling

TheNutritionalValueofFLAVRSAVRTomatoes
TheSafetyofFLAVRSAVRTomatoes

EconomicHarmResultingfromDecreasedConsumptionofTomatoes

CONCLUSIONS

REFERENCES

INTRODUCTION

BackgroundInformation
InthedevelopmentofFLAVRSAVRtomatoes,CalgeneusedrecombinantDNAtechniquestointroducean
antisensepolygalacturonase(PG)gene.ThesensePGgene,normallypresentintomatoes,encodesthe
enzymePG,whichisassociatedwiththebreakdownofpectin(aconstituentofthecellwallintomato
fruit).TheprincipleunderlyingtheFLAVRSAVRtomatoisthattheantisensePGgenesuppressesthe
productionofthePGenzyme.ThissuppressionofthePGenzymeresultsinripefruitthatremainsfirmfor
anextendedperiodandallowsfreshmarkettomatoestoremainonthevinelongerforenhancedflavor.
Thus,FLAVRSAVRtomatoeshavebeenmodifiedtocontainlowerlevelsofanaturallyoccurringenzyme,
whencomparedtoothervarietiesoftomatoes.
InadditiontotheantisensePGgene,FLAVRSAVRtomatoescontainthekanamycinresistancegene(the
kanrgene)thatencodestheenzymeaminoglycoside3'phosphotransferaseII(APH(3')II).Calgeneused
APH(3')IIasaselectablemarkertoidentifyplantcellscarryingtheantisensePGgene.Inanotice
publishedintheFederalRegisterofMay1,1991(56FR20004DocketNo.90A0416),FDAannounced
thatCalgenehadrequestedanadvisoryopinionconcerningwhetherthekanrgenemaybeusedinthe
productionofgeneticallyengineeredtomato,cotton,andrapeseed(i.e.,oilseedrape)plantsintendedfor
humanfoodandanimalfeeduse.Subsequently,CalgenerequestedthatFDAconverttheadvisoryopinion
requesttoafoodadditivepetition.FDAthereafterannounced,intheFederalRegisterofJuly16,1993(58
FR38429)thatCalgenehadsubmitted,andFDAhadfiled,afoodadditivepetition(FAP3A4364)proposing
thatthefoodadditiveregulationsbeamendedtoprovideforthesafeuseofAPH(3')IIasaprocessingaid
inthedevelopmentofnewvarietiesoftomato,oilseedrape,andcotton.FDA'sevaluationofthesafetyof
APH(3')IIisreflectedintheagency'sdecisiononFAP3A4364.
FollowingCFSAN'sevaluationofthedataandinformationsubmittedbyCalgene,andotherrelevant
material,includingpubliccommentsonCalgene'sadvisoryopinionrequest,FDAconvenedapublic
meeting1oftheagency'sFoodAdvisoryCommitteetoundertakeascientificdiscussionofFDA'sapproach
forevaluatingthesafetyofwholefoodsderivedfromanewplantvarietydevelopedusingrecombinant
DNAtechniques.Calgene'sFLAVRSAVRtomatoservedasanexampleoftheagency'sapproach,and
wasthefocusofthecommittee'sdiscussion.
Themembershipofthestandingcommitteewassupplementedwithtemporarymembersandconsultants
tothecommitteerepresentingscientificdisciplinesappropriatetotheevaluationoffoodsderivedfrom
newplantvarietiesdevelopedusingrecombinantDNAtechniques.Atthatmeeting,committeemembers
expressedtheirviewthattherelevantscientificissuesconcerningFLAVRSAVRtomatoeshadbeen
addressedandthattherewasnoreason,fromasafetystandpoint,toprecludeCalgenefrommarketing
FLAVRSAVRtomatoes.Atranscriptofthatmeeting(Ref.1),andacopyofthebackgroundinformation
providedtothecommittee(Ref.2),ispubliclyavailableatDocketsManagementBranch.

ApproachtotheEvaluationofFLAVRSAVRTomatoes

Asdiscussedinthe1992policystatement,thesafetyassessmentoffoodderivedfromanewplantvariety
mayinclude:(1)anevaluationofthepurposeorintendedtechnicaleffectofthegeneticmodification(2)
anevaluationofthesource,identity,function,andstabilityofintroducedgeneticmaterial(3)analytical
studiestodeterminewhetherthegeneticmodificationhadanyeffectsonthecompositionofthefood(such
asthelevelsofimportantnutrientsandnaturallyoccurringtoxicants)and(4)anevaluationofthesafety
ofnewormodifiedsubstances(i.e.,proteins,carbohydrates,andfatsoroils)inthefood.Alsoas
discussedinthatpolicystatement,animalfeedingstudiesorothertoxicologicaltestsarewarrantedonly
whenthecharacteristicsoftheplantorthenatureofthemodificationraisesafetyconcernsthatcannotbe
resolvedbyanalyticalmethods.2
FLAVRSAVRtomatoeshavebeenmodifiedbyareductioninthelevelsofanaturallyoccurringenzyme
thatdegradespectinandtheadditionofanewprotein,APH(3')II,encodedbythekanrgene.Basedon
thesemodifications,webelievethatasafetyevaluationofFLAVRSAVRtomatoesisproperlyaddressed
byananalysisofthefollowinginformation:(1)thesource,identity,function,andstabilityofgenetic
materialintroducedintoFLAVRSAVRtomatoes(2)analyticalstudiesonthecompositionofFLAVR
SAVRtomatoesand(3)thesafetyofAPH(3')II(theproteinproductofthekanrgene).
Overall,weevaluatedthedataandinformationprovidedbyCalgenetodeterminewhetherFLAVRSAVR
tomatoeshavebeensignificantlyaltered,withinthemeaningof21CFR170.30(f)(2),whencomparedto
varietiesoftomatoeswithahistoryofsafeuse.Inotherwords,wedeterminedwhetherFLAVRSAVR
tomatoesareassafeasothercurrentlyconsumedtomatoes.
Inadditiontofoodsafetyissues,weevaluatedenvironmentalconsiderationsassociatedwithFLAVR
SAVRtomatoes.

OverviewofthisMemorandum
Calgeneinitiallyprovideddataandinformationoneighttransgenictomatolines.Calgenesubsequently
submittedadditionalinformationontwoothertransgenictomatolines(designatedCR3613andCR3623)
thatthefirmcurrentlyconsiderstobemorelikelycandidatesfordirectcommercializationorforfurther
developmentintocommercialvarieties(Refs.3and4).Thismemorandumsummarizesthedataand
informationsuppliedbyCalgeneconcerningFLAVRSAVRtomatolinesCR3613andCR3623,the
conclusionsreachedbyCalgene,andthecommentsofagencyscientistsastowhetherFLAVRSAVR
tomatoeshavebeensignificantlyaltered,withinthemeaningof21CFR170.30(f)(2),whencomparedto
varietiesoftomatoeswithahistoryofsafeuse.
Becauseneitherthecharacteristicsoftomatoesnorthenatureofthemodificationraisesafetyquestions
thatcannotberesolvedbyanalyticalmethods,wehavedeterminedthatanimalfeedingstudiesarenot
necessarytoevaluatethesafetyofFLAVRSAVRtomatoes.However,becauseCalgenesubmittedtoFDA
datafromanimalgavagestudies,thismemorandumalsodiscussesthosestudies.
Inaddition,thismemorandumdiscussesenvironmentalconsiderationsassociatedwithFLAVRSAVR
tomatoesandrequirementsforlabelingFLAVRSAVRtomatoes.Italsodiscussesthepubliccommentson
Calgene'sadvisoryopinionrequest,aswellascommentscontainedinacitizenpetition(DocketNo.92P
0222/CP1)filedinaccordancewith21CFR10.30.

FOODSAFETYASSESSMENTCONDUCTEDBYCALGENE

SummaryofDataSubmittedbyCalgene
Calgeneprovidedthefollowingdataandinformationinsupportofthefirm'ssafetyassessmentofFLAVR
SAVRtomatoes:(1)Backgroundinformationontomatoes,includingthelevelofnutrientsandthelevel
ofthenaturallyoccurringglycoalkaloidtoxicant,tomatine(2)dataandinformationdescribingthe
intendedtechnicaleffectoftheantisensePGgene(3)backgroundinformationonthegenetransfer
systemusedtodeliverandincorporatethekanrandantisensePGgenesintotomatoes(4)information
describingthesourcesofgeneticmaterialtransferredtotomatoes(5)dataandinformationconcerning
themolecularstabilityoftheinsertedkanrandantisensePGgenes(6)datacomparingthelevelsof
nutrientsinFLAVRSAVRtomatoestothelevelsintheparentalvariety(7)datacomparingthelevelsof
toxicantsinFLAVRSAVRtomatoestothelevelsinothercommercialvarieties(8)dataandinformation
concerningAPH(3')IIencodedbythekanrgeneand(9)resultsofthree28dayanimalstudiesinwhich
ratsweregavagedwithwater,nontransgenictomatoes,orFLAVRSAVRtomatoes.Thisinformation,

includingtheliteraturereferencessuppliedbyCalgene,isonpublicdisplayatDocketsManagement
Branch.3

BackgroundInformationonTomatoes
Calgenesubmittedseveralliteraturereferencesprovidingbackgroundinformationontomatoes(Ref.5).
MostcommonlycultivatedtomatovarietiesbelongtothespeciesLycopersiconesculentum.Lycopersicon
speciesaremembersoftheSolanaceae(nightshade)family.Tomatoeshavealonghistoryofsafeuseas
food.TomatoesareconsumedraworprocessedandcontributesignificantamountsofvitaminAand
vitaminCtotheU.S.diet.Commercialtomatoesgenerallyarepickedfromthevineatamaturegreen
stage(i.e.,beforetheyarefullyripe)andsubsequentlygassedwithexogenousethylene(asubstancethat
alsooccursnaturallyintomatoesandpromotesripening)toinducearedcolor.
Anaturallyoccurringglycoalkaloidtoxicant,tomatine,occursatlowlevelsindomesticatedtomato
varietiesnondomesticatedspeciesgenerallycontainhigherconcentrationsoftomatinethantomatoes
thatarecultivatedforfooduse.Tomatineisdistributedthroughoutthetomatoplantbutismost
concentratedinleavesandinopeningflowers.Thetomatineconcentrationintomatofruitdependsonthe
degreeofripenessofthefruit,anddeclinesasgreenfruitripenstoyieldredfruit.

TheIntendedEffectoftheAntisensePGgene
FLAVRSAVRtomatoeshavebeenmodifiedtocontainanantisensePGgene.ThesensePGgene,
normallypresentintomatoes,encodestheenzymePG,whichisassociatedwiththebreakdownofpectin
(aconstituentofthecellwallintomatofruit).TheprincipleunderlyingtheFLAVRSAVRtomatoisthat
theantisensePGgenesuppressestheproductionofthePGenzyme,therebypreventingpectin
degradation,whichdelaysthesofteningofripetomatofruitsandallowsfreshmarkettomatoestoremain
onthevinelongerforenhancedflavor.
Thisintendedeffectdoesnotraisesafetyquestions,becausepectiniswidelyconsumedasacomponentof
manyfruitsandvegetablesandalsoisagenerallyrecognizedassafe(GRAS)substancethatisdirectly
addedtomanyfoodproductsasagellingagent(e.g.,inmarmaladeandjelly)orasastabilizer(e.g.,in
beveragesandicecream).Inaddition,asdiscussedbelow,thepreventionofpectindegradationinFLAVR
SAVRtomatoesoccurswithoutadverselyaffectingothercharacteristicsoftomatofruits,suchasnutrient
andtoxicantlevels.

BackgroundInformationontheGeneTransferSystem
CalgenetransformedtheparentalCR3tomatolineusingtheAgrobacteriumtumefaciensgenetransfer
system.CalgenesubmittedseveralliteraturereferencesprovidingbackgroundinformationontheuseofA.
tumefaciensasagenetransfervehicle(Ref.6).Therelevantliteratureissummarizedbelow.
TheabilityofA.tumefacienstoeffectthetransferofDNAintoplantcellsisrelatedtothemechanismby
whichA.tumefacienscausescrowngalltumors(crowngalls)inplants.Thismechanismisassociatedwith
thepresenceinA.tumefaciensofalargeplasmidcalledtheTi(Tumorinducing)plasmid.Crowngallcells,
incontrasttonormalplantcells,containaspecificportionoftheTiplasmid(knownastheTDNA)
integratedintotheplantgenome.TheTDNAcontainsatleasttwoclassesofgenes:(1)oncogenes,which
areresponsiblefortumorformationand(2)geneswhichareresponsibleforsynthesisofopinesinthe
transformedcellsofthecrowngall.4TheTiplasmidalsocontainsvir(virulence)genes,locatedoutside
theTDNAregion,whicharenecessaryforbothexcisionoftheTDNAfromtheTiplasmidandintegration
oftheTDNAintotheplantgenome.Virulencegenesdonotnormallyintegrateintotheplantgenome.
TypicalproceduresforusingAgrobacteriumasaDNAtransfervehicleinvolve"disarming"5the
AgrobacteriumbyremovingtheTDNAregion(andthustheoncogenesandopinesynthesisgenes)from
theTiplasmidandsubsequentlyintroducingaseparateplasmid(calledabinaryplasmid)thatcontainsa
syntheticTDNAregion.Duringconstructionofthebinaryplasmid,theoncogenesandopinesynthesis
genesareremovedfromtheTDNAregionandreplacedwithcloningsitesintowhichthegene(s)of
interestmaybeinserted.AbinaryplasmidisusuallydesignedtobeabletoreplicateinAgrobacteriumas
wellasanintermediatehostsuchasEscherichiacoli.Growingtheplasmidintheintermediatehost
providesplasmidDNAinsufficientquantitytocharacterizetheTDNAregionpriortotransferofthebinary
plasmidtothedisarmedAgrobacteriumstrainthatwilleffectthetransferoftheTDNAtotheplant
genome.

SourcesofDNAIntroducedintoFLAVRSAVRTomatoes

CalgenecreatedadisarmedA.tumefaciensstraincontainingabinaryplasmidwiththekanrandantisense
PGgenes,accompaniedbyappropriateregulatorysequences,insertedintotheTDNAregion.Calgene
transformedCR3tomatocellsusingthisdisarmedA.tumefaciensstrainandregeneratedthetransformed
tomatocellsintomatureplantsthatbecamethefirstgenerationoftomatolinesCR3613andCR3623.
CalgeneprovidedinformationonthesourceandnucleotidesequenceofgeneticmaterialpresentintheT
DNAregionthatwasintroducedintoFLAVRSAVRtomatolinesCR3613andCR3623.Thegenetic
materialpresentintheTDNAregionwasderivedfromA.tumefaciens,E.coli,CauliflowerMosaicVirus,
andtomato.CalgenedescribedthefulllengthkanrandantisensePGgenes,aswellasincompletegenes
presentintheTDNAregion.Calgeneconcludedthatthepotentialfortheexpressionoftheincomplete
genesintroducedintoFLAVRSAVRtomatoesisverylow.
Calgenealsodesignedanexperiment(usingtheSouthernblottechnique)intendedtodemonstratethatthe
genomeofCR3613FLAVRSAVRtomatoesdidnotincorporatesequencesfromthebinaryplasmidother
thanthoselocatedwithintheTDNAregion,andconcludedthatonlysequenceslocatedwithintheTDNAof
thebinaryplasmidhadbeentransferredtotomatolineCR3613.Calgenedidnotprovidesimilar
experimentalevidenceforCR3623FLAVRSAVRtomatoes.Thefirmdidstatethattheyhadimplemented
aproceduretoscreennewlytransformedtomatolines,aspartofanoverallqualityassuranceprogram,
forthepresenceofDNAlocatedoutsidetheTDNAregion.Calgenefurtherstatedthattheywouldnot
commercializeFLAVRSAVRtomatolinescontainingDNAfromoutsidetheTDNAregionunlesstheyfirst
evaluatedthesafetyofthetransferredsequences(Ref.3).
CFSANscientistscommentedontheinformationprovidedbyCalgene,andtheconclusionsreachedby
Calgene,concerningthesourcesofgeneticinformationinFLAVRSAVRtomatoes,asfollows:The
informationsuppliedbyCalgenesatisfactorilycharacterizesthegeneticcompositionoftheTDNAregion
introducedintoFLAVRSAVRtomatoes.TheTDNAregiondoesnotappeartocontainfulllengthgenes
thatwouldencodeanyproteinknowntobeharmful.TheuseofadequatelycontrolledSouthernblotsis
acceptableasascreeningproceduretodetectDNAsequencesderivedfromthesequencesofabinary
plasmidlocatedoutsidetheTDNA.

GeneticStability
CalgeneprovideddataonthegeneticstabilityofthetransferredDNAinsuccessivegenerationsofCR3613
FLAVRSAVRtomatoes.CalgeneanalyzedtheabilityoftheCR3613linetomaintainaspecificDNA
fragmentconsistingofaborderbetweentheinsertedTDNAregionandthetomatoDNA,andconcluded
thattheinsertedDNAcontainingthekanrandantisensePGgenesremainedstablyincorporatedinthe
tomatogenomeoverthecourseoffivegenerations.CalgenedidnotsubmitsimilardataforCR3623
FLAVRSAVRtomatoes.
CFSANscientistscommentedontheinformationprovidedbyCalgene,andtheconclusionsreachedby
Calgene,concerningthegeneticstabilityofFLAVRSAVRtomatolineCR3613,asfollows:The
informationsuppliedbyCalgeneestablishesthattheinsertedTDNAisstablyintegratedintoFLAVR
SAVRtomatolineCR3613andthatthestructureoftheinsertedTDNAinFLAVRSAVRtomatoline
CR3613remainsunchangedoverfivegenerations.Asapracticalmatter,stableintegrationofatraitina
commercialplantvarietyisinherentinthedevelopmentoftheplantandanyinstabilitywouldlikelybe
discoveredinqualitycontroltestsroutinelyconductedbyplantbreederswhenmaintainingproduction
varieties.

NutrientsinFLAVRSAVRTomatoes
CalgenecomparedthenutritionalprofileofFLAVRSAVRtomatolinesCR3613andCR3623tothe
nutritionalprofileoftheparental(CR3)varietyoveraperiodoftimecorrespondingtotheexpectedshelf
lifeofvineripenedfreshmarkettomatoes(i.e.,tomatoespickedpink)andgassedgreentomatoes(i.e.,
tomatoespickedatamaturegreenstageandgassedwithethylene).Maturegreenandpinkfruitsfrom
boththeFLAVRSAVRandparentallineswereharvestedonthesameday,andthegreenfruitswere
gassedwithethylene.RepresentativefruitswereanalyzedperiodicallyforvitaminAandvitaminClevels
duringstorageunderconditionsexpectedforcommercialtomatoes.Thelastanalysiswasperformedat
theendoftheshelflife(i.e.,after25daysfortomatoespickedmaturegreenandafter18daysfor
tomatoespickedpink).Calgenereportedthat,duringthecourseoftheirevaluation,meanvaluesobtained
forlevelsofvitaminAandvitaminCinfruitsobtainedfromboththeFLAVRSAVRandparentallines
wereallwithintherangereportedintheliterature.Calgeneconcludedthatnosignificantdifferencesexist
betweentheFLAVRSAVRtomatolinesandthecontrolparentaltomatolinewithrespecttolevelsof
vitaminAandvitaminC.

CFSANscientistscommentedontheinformationprovidedbyCalgene,andtheconclusionsreachedby
Calgene,concerninglevelsofvitaminAandvitaminCinFLAVRSAVRtomatoes,asfollows:Thelevels
ofbothnutrientsvariedamongFLAVRSAVRfruitsaswellasamongcontrolfruits.Suchvariabilitycould
reflectdifferencesinfieldconditions,weather,postharvesttreatment,aswellasotherfactors.
Consideringthisoverallvariability,agencyscientistsnotednosignificantdifferencesinnutrientlevels
betweenFLAVRSAVRtomatoesandcontroltomatoes,andconsideredthatalllevelsreportedforvitamin
AandvitaminCwerewithintherangeregardedasnormal(asreflectedinliteraturedata).6

GlycoalkaloidToxicantsinFLAVRSAVRTomatoes
CalgenecomparedthetomatinelevelsinmaturegreenandredripeFLAVRSAVRtomatolinesCR3613
andCR3623tothetomatinelevelsinmaturegreenandredripecommercialtomatovarieties.Calgene
reportedthattomatinelevelsingreenFLAVRSAVRtomatoeswerecomparabletotomatinelevelsinone
commercialtomatovariety,butsomewhathigherthantomatinelevelsinasecondcommercialtomato
variety.Calgenedetectedtomatine(atalimitofdetectionof2.5ppm)inonlyoneoutof38redripe
FLAVRSAVRfruitsandfouroutof60redripecommercialfruits.Calgeneconcludedthat,asexpected,
tomatinelevelsdecreasedduringtomatoripeninginbothFLAVRSAVRandcommercialtomatolinesand
thattomatinelevelsinredripefruit(i.e.,tomatoesastheywillbepresentedtoconsumers)ofFLAVR
SAVRtomatolinesarecomparabletotomatinelevelsinredripefruitofcommercialtomatovarieties.
Calgeneusedahighperformanceliquidchromatography(HPLC)methodtomeasuretomatinelevels.
Calgeneanalyzedtheunidentifiedpeakshavingelutiontimeswithintherangeexpectedforglycoalkaloids
bycomparingtheirelutiontimesandultravioletspectratothoseofmajorglycoalkaloidsthatoccurin
potatoes.Basedontheseanalyses,Calgenedeterminedthattheunidentifiedpeaksdonotrepresent
glycoalkaloidsandconcludedthatnosignificantdifferencesinthecontentofglycoalkaloidswereobserved
betweenFLAVRSAVRandcommercialtomatovarieties.
CFSANscientistscommentedontheinformationprovidedbyCalgene,andtheconclusionsreachedby
Calgene,concerninglevelsoftomatineandotherglycoalkaloidsinFLAVRSAVRtomatoes,asfollows:
TheinformationsuppliedbyCalgenesupportsCalgene'sobservationthatthetomatinelevelsinvine
ripenedFLAVRSAVRtomatoesarecomparabletothetomatinelevelsincommercialtomatovarieties.
Calgene'sdataalsoprovideevidencethatneitherFLAVRSAVRtomatoesnorcommercialcontroltomato
varietiescontainotherglycoalkaloidsatlevelsmeasurablewiththeHPLCmethodused.

NewProteinsIntroducedintoFLAVRSAVRTomatoes
Asnotedabove,indevelopingFLAVRSAVRtomatoes,CalgeneusedAPH(3')IIasaselectablemarkerto
identifyplantcellscarryingtheantisensePGgene.Calgeneprovideddataandinformationaddressingthe
safetyofAPH(3')II,includingthedirecteffectsofingestionaswellaseffectsassociatedwiththebiological
activityofAPH(3')II(i.e.,theeffectoftheenzymeonthetherapeuticefficacyoforallyadministered
antibiotics).FDAhasevaluatedthesafetyofAPH(3')IIinthecontextofafoodadditivepetition(FAP
3A4364)andhasconcludedthatAPH(3')IIissafeforuseasaprocessingaidinthedevelopmentof,
amongotherplants,tomatoes.

AnimalGavageStudies

Introduction
WhenCalgeneapproachedFDAin1991,whiletheagencywasdevelopingthe1992policystatement,the
firmindicatedthattheywishedtoprovideadditionalassurancethatallpossibleteststoestablishthe
safetyofthisfirstexampleofafoodderivedfromanewplantvarietydevelopedusingrecombinantDNA
techniqueshadbeenperformed.Onequestionraisedwaswhethertoxicologicalstudiescouldhelpto
determinewhetherunexpectedtoxicantsmightbepresentinFLAVRSAVRtomatoes.Althoughthereare
nowellestablishedtoxicologicalapproachestotestingthesafetyofwholefoods,Calgeneacceptedan
agencysuggestion(Refs.7and8)thatthefirmconductashorttermfeedingstudyinrodents.Thefirm
thendesignedandconductedthreeratgavagestudies.
Results
Calgenesubmitteddatafromthreeshortterm(28day)gavagestudies,conductedattheInternational
ResearchandDevelopmentCorporation(IRDC),inwhichgroupsofmaleandfemaleratsweregiven(by

gavage)deionizedwater,control(nontransgenic)tomatoes,orFLAVRSAVRtomatoes.IRDCidentifiedno
biologicallysignificantchangesinbodyweight,organweight,foodconsumption,hematologicparameters
orclinicalchemistryfindingsinanyofthethreestudies.Inaddition,inthefirststudy,IRDCidentifiedno
adversefindingsthatcouldberelatedtoconsumptionofFLAVRSAVRtomatoesfollowingcompletegross
andmicroscopicexaminationofacomprehensiveselectionoftissues.Inthesecondandthirdstudies,
however,IRDCnotedgastricerosionsinsomeanimals.Thesefindingsonthegastricerosionsare
discussedbelow.
GastricErosions
Inthefirststudy,groupsofmaleandfemaleratsweregivenbygavageeither(1)deionizedwater(2)
homogenizedFLAVRSAVRtomatoesobtainedfromanoncommercialtomatolineor(3)homogenized
nontransgenictomatoes.IRDCreportednogastricerosionsinratsfromanygroup.(SeeTable1.)
Inthesecondstudy,groupsofmaleandfemaleratsweregivenbygavageeither(1)deionizedwater(2)
homogenizednontransgenicCR3tomatoes(3)homogenizedFLAVRSAVRtomatoesobtainedfromthe
CR3613tomatolineor(4)homogenizedFLAVRSAVRtomatoesobtainedfromtheCR3623tomatoline.
Inthissecondstudy,IRDCreportedgastricerosionsinfouroftwentyfemaleratsgivenCR3623FLAVR
SAVRtomatoes,butnotinratsinanyothergroup.(SeeTable1.)
Inthethirdstudy(whichwasdesignedinanattempttoclarifytheresultsofthesecondstudy),groupsof
maleandfemaleratsweregivenbygavageeither(1)deionizedwater(2)homogenizednontransgenic
CR3tomatoes(3)lyophilizednontransgenicCR3tomatoes7(4)homogenizedCR3623FLAVRSAVR
tomatoes(5)lyophilizedCR3623FLAVRSAVRtomatoesgrowninonegeographicallocationor(6)
lyophilizedCR3623FLAVRSAVRtomatoesgrowninasecondgeographicallocation(femalesonly).In
thisthirdstudy,IRDCreportedgastricerosionsineightofelevengroups.(SeeTable1.)
AttherequestofCalgene,PATHCO,Inc.,assembledapanelofpathologists(thePathologyWorkingGroup
orPWG),whoconductedareviewofcodedmicroscopicslidescontainingstomachsectionsfromallthree
studiestoevaluatetheincidenceandsignificanceoftheobservedgastricerosions.(SeeTable1.)Alsoat
therequestofCalgene,ENVIRONCorporationpreparedasummaryofCalgene'soverallsafetyassessment
ofFLAVRSAVRtomatoesandassembledanexpertpaneltoreviewthatsummary.ThePWGconcluded,
andtheexpertpanelconcurred,thatthegastricerosionsobservedwereincidentalandnottestarticle
related.CalgenesubmittedtoCFSANtheoriginaldatafromtheIRDCstudies,thePWGreportonthethree
gavagestudies,andtheconclusionsoftheexpertpanel.
TABLE1
IncidenceofRatswithGastricErosions
TreatmentGroup

IncidenceofRatswithGastricErosions8
Sex
Study1
Study2
Study3
9
10 IRDC PWG IRDC PWG
IRDC PWG

M
F
M
Nontransgenic(VickieMale)controltomatoes(fresh)
F
M
FLAVRSAVRtomatoline501100115(fresh)
F
M
NontransgenicCR3controltomatoes(frozen)(Dixon)11
F
M
NontransgenicCR3controltomatoes(lyophilized)(Indio)
F
M
FLAVRSAVRtomatolineCR3613(frozen)
F
M
FLAVRSAVRtomatolineCR3623(frozen)(Dixon)
F
M12

0/20
0/20
0/20
0/20
0/20
0/20

0/20
0/20
0/20
0/20
1/20
0/20

0/20
0/20

0/19
0/20

0/20
0/20
0/20
4/20

0/20
0/20

0/19
0/20

1/20
1/20
1/20
7/20

3/20
1/20

3/20
2/20
1/20
0/19

0/20
3/20
0/20

4/20
1/20

3/20
2/20
1/20
0/20

0/20
1/20
0/20

F13

1/20

0/20

F14

2/15

2/15

Water

FLAVRSAVRtomatolineCR3623(lyophilized)

Calgenesubsequentlysubmittedadditionaldataandinformationonthegastricerosions,includinga

PathologyConsensusReport(preparedbythepathologistsfromIRDCandPWG),historicalcontroldata,
andselectedphotomicrographsofthegastricerosionsfromCalgene'sgavagestudies.ThePathology
ConsensusReportdescribedthegastricerosionsasbeingminimaltomildinseveritystatedthatthe
morphologyandseverityofgastricerosionsweresimilarinratsthatreceivedtransgenictomatoes,non
transgenictomatoes,ordeionizedwaterandpointedoutthatnodoseresponserelationshipwasobserved
whenthedoseoftomatoeswasdoubled.Thereportalsonotedthatgastricerosionsseeninthesestudies
"...arenotuniquelesionssincetheycanalsobecausedbyphysiologicfactorssuchasstressorfastingas
wellasbyawidevarietyofagentsincludingdrugs,chemicals,andnaturaltoxinssuchastomatine."In
thejudgmentofPWGandIRDCpathologists,thelesionswereofshortdurationandwouldlikelybe
completelyresolvedinashorttime.Calgene'sPathologyConsensusReportstatedthat"[a]llofthe
pathologistsinvolvedinthereviewofthestudiesonthetransgenictomatoesconcludedthattherewereno
effectsobservedintheglandularstomachwhichwererelatedtotheadministrationoftransgenictomatoes
toSpragueDawleyratsbygavage."
Conclusions
GiventhenatureofthemodificationtoFLAVRSAVRtomatoes(i.e.,thesuppressionoftheproductionof
thenaturallyoccurringPGenzymeandtheadditionofAPH(3')IIencodedbythekanrgene),CFSAN
believes,asnotedabove,thatasafetyevaluationofFLAVRSAVRtomatoesisproperlyaddressedbyan
analysisofthefollowinginformation:(1)thesource,identity,function,andstabilityofgeneticmaterial
introducedintoFLAVRSAVRtomatoes(2)analyticalstudiesonthecompositionofFLAVRSAVR
tomatoesand(3)thesafetyofAPH(3')IIencodedbythekanrgene.Asdiscussedabove,Calgenehas
providedsuchinformation,whichhasservedasthebasisofthefirm'sevaluation.Inlightofthis
information,CFSANhasdeterminedthatdatafromanimalgavagestudiesarenotnecessarytoan
evaluationofwhetherFLAVRSAVRtomatoesaresignificantlyaltered,whencomparedtoothertomatoes
withahistoryofsafefooduse.
However,Calgenedidconductanimalgavagestudiesandsubmitthemtotheagencyforevaluation.
CFSANscientistsdidcommentontheinformationprovidedbyCalgene,andtheconclusionsreachedby
Calgene,concerningthesestudies,asfollows:Thethreestudiesconsistentlydemonstratednobiologically
significantchangesinbodyweight,organweight,foodconsumption,hematologicparametersandclinical
chemistryfindings.Therewasdisparityamongthethreestudiesregardingtheincidenceofratswith
gastricerosions.DataandinformationsuppliedbyCalgenefailtoclarifyorexplainthefactorsresponsible
forthisdisparity.BasedontheinformationCalgenehasprovided,nodefinitiveconclusionscanbedrawn
regardingtheetiology(ies)ofthegastricerosions.Regardlessoftheetiology(ies),however,thegastric
erosionsasdescribedbyCalgenearenomoresevereintransgenictomatoesthaninnontransgenic
tomatoes.

STATUSOFFLAVRSAVRTOMATOES
Insum:
1. TheintendedeffectoftheantisensepolygalacturonaseRNAinFLAVRSAVRtomatoesdoesnotraise
safetyquestions.
2. TheTDNAtransferredtoFLAVRSAVRtomatoesdoesnotappeartocontainfulllengthgenesthat
wouldencodeanyproteinknowntobeharmful.
3. TheinformationsuppliedbyCalgeneestablishesthattheinsertedTDNAisstablyintegratedinto
FLAVRSAVRtomatolineCR3613andthatthestructureoftheinsertedTDNAinFLAVRSAVR
tomatolineCR3613remainsunchangedoverfivegenerations.
4. ThelevelsofvitaminAandvitaminCinFLAVRSAVRtomatolinesCR3613andCR3623are
comparabletothelevelsofthesesamenutrientsincontrol(parental)tomatoes.Alllevelsreported
forthesenutrientswerewithintherangeconsiderednormal,asreflectedinliteraturedata.
5. TomatinelevelsinvineripenedFLAVRSAVRtomatolinesCR3613andCR3623arecomparableto
tomatinelevelsincommercialtomatovarieties.Calgene'sdataprovideevidencethatneitherFLAVR
SAVRtomatoesnorcommercialcontroltomatovarietiescontainotherglycoalkaloidsatlevels
measurablewithanHPLCmethod.
6. ThesafetyofanewproteininFLAVRSAVRtomatoes,APH(3')II,isthesubjectofFAP3A4364.In
thatrulemaking,afairevaluationofthedataavailabletoFDAestablishesthatAPH(3')IIissafefor
useasaprocessingaidinthedevelopmentof,amongotherplants,newvarietiesoftomatoes.
7. AdditionalinformationsubmittedbyCalgene(i.e.,thedatafromthe28dayratgavagestudies)are

notnecessarytoourdeterminationofwhetherFLAVRSAVRtomatoesareassafeasother
currentlyconsumedtomatoes.Nevertheless,thethreestudiesconsistentlydemonstratedno
biologicallysignificantchangesinbodyweight,organweight,foodconsumption,hematologic
parametersandclinicalchemistryfindings.Therewasdisparityamongthethreestudiesregarding
theincidenceofratswithgastricerosions.BasedontheinformationCalgenehasprovided,no
definitiveconclusionscanbedrawnregardingtheetiology(ies)ofthegastricerosions.Regardlessof
theetiology(ies),however,thegastricerosionsasdescribedbyCalgenearenomoreseverein
transgenictomatoesthaninnontransgenictomatoes.
BasedontheinformationCalgenesubmittedconcerningFLAVRSAVRtomatoes,wehavedeterminedthat
thisnewvariety,theFLAVRSAVRtomato,hasnotbeensignificantlyalteredwithinthemeaningof21
CFR170.30(f)(2),whencomparedtovarietiesoftomatoeswithahistoryofsafeuse.Inotherwords,the
FLAVRSAVRtomatoisassafeasothercommonlyconsumedtomatoes.

ENVIRONMENTALCONSIDERATIONS
Assetforthinthe1992policystatement(57FR22984at23005),aconsultation,suchastheconsultation
betweenCalgeneandFDAonFLAVRSAVRtomatoes,isnotanagencyactionundertheNational
EnvironmentalPolicyAct(NEPA).Therefore,neitheranenvironmentalassessment(21CFR25.22)nora
claimforacategoricalexclusion(21CFR25.23and21CFR25.24)isrequired.15
CalgenepetitionedtheU.S.DepartmentofAgricultureAnimalandPlantHealthInspectionService(APHIS)
andrequestedadeterminationoftheregulatorystatusofFLAVRSAVRtomatoesunderthePlantPest
Act.APHISdeterminedthatFLAVRSAVRtomatoes:(1)exhibitnoplantpathogenicproperties(2)areno
morelikelytobecomeaweedthannonengineeredparentalvarieties(3)areunlikelytoincreasethe
weedinesspotentialforanyothercultivatedplantornativewildspecieswithwhichtheorganismcan
interbreed(4)donotcausedamagetoprocessedagriculturalcommoditiesand(5)areunlikelytoharm
otherorganismsthatarebeneficialtoagriculture.BecauseFLAVRSAVRtomatoesdonotpresentaplant
pestriskandarenototherwisedeleterioustotheenvironment,APHISdeterminedthatFLAVRSAVR
tomatoes,previouslyfieldtestedunderpermit,wouldnolongerbeconsideredregulatedarticles(57FR
47608October19,1992).
However,CalgenesubmittedtoFDAacopyofthefirm'spetitiontoAPHIS.CFSANscientistsreviewedthe
informationprovidedbyCalgeneandconcurwithAPHIS'conclusionsregardingtheenvironmentalsafetyof
FLAVRSAVRtomatoes.

LABELING

StatutoryandRegulatoryRequirementsforLabeling
Section403oftheactgovernsthelabelingoffood.Undersection403(a)(1),afoodismisbrandedifits
labelingisfalseormisleading.Undersection201(n)oftheact,labelingismisleadingifitfailstoreveal
allfactsthatare"materialinlightof***representations[madeorsuggestedinthelabeling]ormaterial
withrespecttoconsequenceswhichmayresultfromtheuseofthearticletowhichthelabeling***
relatesundertheconditionsofuseprescribedinthelabeling***orundersuchconditionsofuseasare
customaryorusual."
Section403(i)oftheactandregulationspromulgatedthereunder(21CFR101.3)requirethatafood
productbedescribedbyitscommonorusualnameor,intheabsencethereof,anappropriatelydescriptive
term.Section403(i)oftheactalsorequiresthat,inthecaseoffoodsfabricatedfromtwoormore
ingredients,afoodproductbearonthelabelthecommonorusualnameofeachingredient.

LabelingFoodsDerivedfromNewPlantVarieties
The1992policystatementdescribedsituationsinwhichadeveloperofanewplantvarietyshouldconsult
FDAtodeterminewhetherspeciallabelingwouldberequiredforthefood.Examplespotentiallyrelevantto
FLAVRSAVRtomatoesinclude(1)thealterationoflevelsofimportantnutrientstolevelsnotwithinthe
rangeordinarilyseeninthehostspeciesand(2)theexistenceofasafetyorusageconcern,suchasthe
introductionintothefoodofaproteinderivedfromacommonlyallergenicfoodifthereisinsufficient
informationtodemonstratethattheallergenicdeterminanthasnotbeentransferred.
Inthe1992policystatement,FDAalsostatedthattheagencydoesnotbelievethatthemethodof

developmentofanewplantvariety(includingtheuseofnewtechniquessuchasrecombinantDNA)is
normallymaterialinformationwithinthemeaningofsection201(n)andwouldnotusuallyberequiredto
bedisclosedinlabeling.Inlightofcommentsreceivedtothe1992policystatement,FDApublisheda
notice(the1993labelingnotice)requestingdataandinformationonseveralissuesrelatingtothelabeling
ofgeneticallyengineeredfoods,includingwhetherallgeneticallyengineeredfoodsshouldbelabeledto
revealthatfacttoconsumers(58FR25837April28,1993).Weareintheprocessofevaluatingthe
commentsreceivedinresponsetothatnotice.Todate,wehaveencounterednofactualinformationinthe
commentsthatwouldprovidethebasistoalterFDA'scurrentinterpretationoftheact,underwhichspecial
labelingfortheclassofgeneticallyengineeredfoodsisnotrequired.

RequirementsforLabelingFLAVRSAVRTomatoes
WehavereviewedtheinformationsubmittedbyCalgeneconcerningFLAVRSAVRtomatoesinlightof(1)
thestatutoryprovisionsforlabelingandexistingagencyregulationsandpolicyimplementingthose
statutoryprovisionsand(2)theopinionsoftheagencyscientistswhoevaluatedtheinformationsubmitted
byCalgene.WebelievethatthecorrectcommonorusualnamefortheFLAVRSAVRtomatois"tomato",
becausetheFLAVRSAVRtomatohasnotbeensignificantlyalteredwhencomparedtotherangeof
commercialvarietiesreferredtoby"tomato."Wealsofindnosafetyorusageconcerntowhichconsumers
ofFLAVRSAVRtomatoesmustbealertedbyspeciallabeling.(Thelabelingissuesrelatingtothe
presenceofAPH(3')IIinfoods,suchasFLAVRSAVRtomatoes,arediscussedintheagency'sevaluation
ofFAP3A4364.)

EVALUATIONOFCOMMENTS

Overview
FDAreceived20commentstothenoticeannouncingCalgene'srequestforanadvisoryopiniononthe
statusofFLAVRSAVRtomatoes.Commentswerereceivedfrommanufacturers,tradeorganizations,
universities,consumerorganizations,individualconsumersandonestategovernment.Inaddition,a
citizenpetitionrelatingtoFLAVRSAVRtomatoeswasfiledinaccordancewith21CFR10.30.Weaddress
belowthoseissuesraisedinthecitizenpetitionrelevanttoFLAVRSAVRtomatoes,andhaveprepareda
separatereplythatrespondstotheactionsrequestedinthecitizenpetition.
Allofthecommentsfrommanufacturers,tradeorganizations,universities,thestategovernment,andone
commentfromanindividualconsumerexpressedsupportforthemarketingofFLAVRSAVRtomatoes.
SeveralofthesecommentsstatedthatCalgene'sFLAVRSAVRtomatoisidenticaltoothertomatoes,has
beenextensivelyevaluatedandfieldtested,andposesnofoodorenvironmentalsafetyproblems.Several
commentsstatedthatCalgene'sFLAVRSAVRtomatowouldservetointroducethepublictothevalueof
geneticallyengineeredfoodsandinstillpublicconfidenceinFDA'ssciencebasedsafetyassessment.Other
commentsstatedthatFLAVRSAVRtomatoescouldanswerconsumerdemandsfortomatoeswithmore
flavorthantomatoesthatarecurrentlymarketed.Twocommentsspecificallyrespondedtoandsupported
Calgene'sclaimthatFLAVRSAVRtomatoesarefoodsubjecttoacategoricalexclusionfromNEPA.We
seenoneedtofurtherdiscussthesecommentssupportingthemarketingofFLAVRSAVRtomatoes.
CommentsfromtheEnvironmentalDefenseFund,theFoundationonEconomicTrends,andthreeindividual
consumers,aswellascommentscontainedinacitizenpetitionfiledbytheFoundationonEconomic
Trends,opposedthemarketingofFLAVRSAVRtomatoes.Thesecommentsraisedavarietyofissues,
whichwediscussindetailbelow.

RegulatoryIssuesRaisedintheComments

Premarketnotification
ThecitizenpetitionandonecommentrequestedthatFDArequirethatproducersofgeneticallyengineered
foodsanalyzethecompositionofthesefoodsandnotifyFDAoftheresultsoftheseanalysesatleast90
daysbeforethefoodsareintroducedordeliveredforintroductionintointerstatecommerce.Totheextent
thatthesecommentssuggestthatFDArequiremandatorypremarketnotificationforfoodsotherthan
FLAVRSAVRtomatoes,thecommentsdonotrequirearesponseinthecontextofFDA'sconsultation
concerningFLAVRSAVRtomatoes.Further,asdiscussedindetailabove,CalgenehassubmittedtoFDA

analyticaldataonthecompositionofFLAVRSAVRtomatoesandthushaseffectivelygivenFDA
premarketnotificationmorethan90daysinadvanceoftheirintenttocommercializeFLAVRSAVR
tomatoes.Accordingly,webelievethatthecommentsregardingpremarketnotificationrequirenofurther
response.
Premarketapproval
OnecommentassertedthatFDAshouldnotapproveCalgene'stomato,butshouldletconsumeracceptance
serveastheapproval.Calgene'sconsultationwithFDAwillnotresultinanapprovalperseofFLAVR
SAVRtomatoes.Rather,asaresultoftheconsultation,wehavedeterminedthatFLAVRSAVR
tomatoeshavenotbeensignificantlyalteredwithinthemeaningof170.30(f)(2),whencomparedto
varietiesoftomatoeswithahistoryofsafeuse.Thus,consumeracceptancewillstillplayaroleinthe
marketingofFLAVRSAVRtomatoes.
FDAregulationsconcerningadvisoryopinions
OnecommentprovidedthreeargumentsinsupportofitscontentionthatFDAshoulddenyCalgene's
requestforanadvisoryopinionbecausethatrequestviolatedtheletterandspiritofFDAregulations
governingadvisoryopinions(21CFR10.85).TheseregulationsgivetheCommissionerofFoodandDrugs
thediscretiontodenyarequestforanadvisoryopinionif"[t]herequestcoversaparticularproduct***
anddoesnotraiseapolicyissueofbroadapplicability"(21CFR10.85(a)(2)(iv))orif"[t]herequest
containsincompleteinformationonwhichtobaseaninformedadvisoryopinion"(21CFR10.85(a)(2)(i)).
First,thecommentstatedthatCalgene'srequestcoversaparticularproduct.Second,thecomment
assertedthatCalgene'srequestdoesnotraiseanyissuesofbroadapplicabilitythatwerenotraised
independentlyinthe1992policystatement,andthatanypolicyissuesnotadequatelyaddressedinthe
1992policystatementshouldproperlybeaddressedthroughthecommentstothatstatement.Third,the
commentdeclaredthatCalgene'srequestcontainedincompleteinformationbecausethesafetyofthekanr
geneforuseasaselectablemarkeringeneticallyengineeredwholefoodswasstillunderreview.
BecausewehaverecommendedthatFDAnotprovideanadvisoryopinionunder21CFR10.85,webelieve
thatthiscommentrequiresnofurtherresponse.Thatis,weagreethatanadvisoryopinionunder21CFR
10.85concerningthestatusofaspecificfoodproductderivedfromanewplantvarietyisnolonger
appropriateinlightofthepublicationofthe1992policystatement.However,becauseCalgenesubmitted
itsrequestbeforeFDAissuedthe1992policystatement,wedonotagreethatCalgene'srequestis
inconsistentwith21CFR10.85.16
ThestatusofnewsubstancesintroducedintoFLAVRSAVRtomatoes
Commentsassertingthatthekanrgeneanditsexpressionproduct(i.e.,APH(3')II)arefoodadditives
subjecttopremarketapprovalareaddressedbytheagency'sdeterminationinFAP3A4364anddonot
requirearesponseinthismemorandum.(Infact,suchcommentsareessentiallymootinthatCalgene
filed,andtheagencyhasevaluated,afoodadditivepetitionforAPH(3')II.)
OnecommentmaintainedthatCalgenedeliberatelyaddedtheantisensePGRNAtoachievespecific
technicaleffectsinthetomatoandthatthereforetheantisensePGRNAisafoodadditiveunlessitis
GRAS.ThecommentarguedthattheremaynotbeanadequatebasistoconcludethattheantisensePG
RNAisGRASifFDAdoesnotfindthatsufficientscientificstudieshavebeenpublisheddemonstratingthe
safetyoftheantisensePGRNAundertheintendedconditionsofuse.Thecommentarguedthat,although
CalgenesubmittedtoFDAalargenumberofpublishedstudiesconcerningtomatobiologyandgenetic
engineeringtechniques,Calgenehadnotsubmittedapublishedscientificstudyaddressingthetoxicology
andnutritionalvalueofFLAVRSAVRtomatoes.
Webelievethattheissuesraisedinthesecommentswereaddressedinthe1992policystatement.
Specifically,inthatstatementFDAexplainedthattheagencydidnotanticipatethattransferredgenetic
materialwoulditselfbesubjecttofoodadditiveregulation,becausenucleicacidsarepresentinthecells
ofeveryplantusedforfoodbyhumansoranimalsanddonotraiseasafetyconcernasacomponentof
food(57FR22984at22990).Moreover,wenotethatnucleicacidsareefficientlydigested(Ref.9).
FDAalsoexplainedinthe1992policystatementthattheintroductionofageneencodinganantisenseRNA
wouldnotraiseconcernsabouteitherthegeneortheantisenseRNA,andthatanysafetyconsiderations
associatedwithantisenseRNAwouldfocusontheintendedeffectsoftheantisenseRNA(57FR22984at
23004).Asdiscussedabove,thisintendedeffectdoesnotraisesafetyquestions.
Furthermore,wenotethatpublishedstudiesordinarilyarerequiredinthecontextofaGRASaffirmation

petition.17Here,however,CalgenedidnotsubmitaGRASaffirmationpetitionand,thus,FDAhasnotbeen
askedtoaffirmasubstance,suchastheantisensePGRNA,asGRAS.Therefore,webelievethatitisnot
necessarytorequireCalgenetosubmitpublishedstudies.
ThescopeofFDA'sresponse
OnecommentmaintainedthatFDAshoulddenyrulingonCalgene'srequestbecausethescopeofanysuch
rulingwouldbeunclear.ThecommentquestionedwhetherFDA'sresponsewouldencompassFLAVR
SAVRtomatoes,allgeneticallyengineeredtomatoes,orallgeneticallyengineeredfoodswhetherFDA's
responsewouldencompassanyotherwholefoodscontainingthekanrgene,orallwholefoodscontaining
thekanrgeneandwhetherFDA'sresponsewouldencompassgeneticallyengineeredfoodscontaining
multiplecopiesofthekanrgene.
Wenotethatourevaluation,conductedinthecontextofaconsultationwithCalgene,isspecifictoFLAVR
SAVRtomatoesanddoesnotaddressanyothervarietyofgeneticallyengineeredtomatooranyother
geneticallyengineeredwholefood.ThecommentsconcerningthescopeofFDA'sdecisionwithrespectto
thekanrgeneneednotbeaddressedinthismemorandumbecausethescopeofFDA'sdecisionconcerning
thekanrgeneisdiscussedintheagency'sdecisiononFAP3A4364.
Labeling
SeveralcommentsrequestedthatgeneticallyengineeredfoodssuchasFLAVRSAVRtomatoesbelabeled
torevealthatfacttoconsumers.WehavediscussedabovethereasonswhyFLAVRSAVRtomatoesare
notsubjecttoanyspeciallabeling.Moreover,wenotethatCalgenehasannouncedanintentiontoprovide
voluntarilytoconsumerslabelingthatexplainsthatFLAVRSAVRtomatoesweredevelopedusinggenetic
engineeringtechniques(Ref.10).
OthercommentsrequestedthatAPH(3')IIbelabeledasaningredientofFLAVRSAVRtomatoesandof
anyfoodcontainingFLAVRSAVRtomatoes.Asdiscussedintheagency'sdecisionconcerningFAP
3A4364,FDA'sauthorityoverfoodlabelingisbasedonsection403oftheact(21U.S.C.343).Section
403(i)oftheactrequiresthat,inthecaseoffoodsfabricatedfromtwoormoreingredients,afood
productbearonthelabelthecommonorusualnameofeachingredient,unlesscompliancewiththe
requirementforlabelingisimpracticableorresultsindeceptionorunfaircompetition.FDAconsidersan
"ingredient"tobeasubstanceusedtofabricate(i.e.,manufactureorproduce)afood.FDAdoesnot
considerthosesubstancesthatareinherentcomponentsofafoodtobeingredientsthatmustbedisclosed
onthefood'slabel.
Ageneticsubstanceintroducedintoaplantbybreedingbecomesaninherentpartoftheplantaswellasof
allfoodsderivedfromtheplant.ConsistentwithFDA'sgeneralapproachoningredientlabeling,theagency
hasnottreatedasaningredientanewconstituentofaplantintroducedbybreeding,regardlessofthe
methodusedtodevelopthevariety.ThecommentsprovidenobasisforFDAtodeviatefromitscurrent
practiceinthecaseofAPH(3')II.18Accordingly,asdiscussedintheagency'sdecisiononFAP3A4364,FDA
hasdeterminedthatneitherthekanrgenenorAPH(3')IIisaningredientthat,undersection403(i)ofthe
act,mustbeindividuallyidentifiedinlabelsoffoodscontainingthem.

TheNutritionalValueofFLAVRSAVRTomatoes
OnecommentarguedthatCalgene'sanalysisofthenutritionalcompositionofFLAVRSAVRtomatoeswas
inadequatefortworeasons.First,thecommentassertedthatCalgenedidnotstatehowlongafterharvest
thetomatoanalyseswereperformed.ThecommentarguedthatCalgeneshouldcomparethenutritional
valueoffreshmarketFLAVRSAVRtomatoestothenutritionalvalueofnontransformedfreshmarket
tomatoesafteraperiodoftimethatcorrespondstotheirrespectiveshelflives,becauseCalgene's
tomatoesareengineeredtohavealongershelflifeandsomenutrientsmaydegradeovertime.In
addition,thecommentarguedthatCalgeneshouldobtainnutritionaldataappropriatetoprocessing
tomatoes(i.e.,soonafterharvest).Second,thecommentassertedthatCalgenedidnotuseformal
statisticalhypothesistestingwhendesigningthefirm'sexperimentscomparingnutritionaldataforFLAVR
SAVRtomatoesandparentalvarieties,andnotedthatCalgenedidnothaveevenclosetoa50percent
chanceofdetectinga20percentdifferenceinnutritionalvaluebetweenFLAVRSAVRtomatoesandthe
parentalvariety.Thecommentciteda1974presentationbyanFDAofficialatanannualmeetingofa
tradeorganizationinwhichtheofficialindicatedthattheagencywouldconsidera20%differencein
nutritionalcontenttobeasignificantchangeinthenutritionalvalueoffoodderivedfromacropplant
(Ref.11).

Calgeneoriginallyprovidedlimitednutrientcompositiondatainsupportoftheirsafetyassessmentofeight
FLAVRSAVRtomatolinesthatthefirmisnotcurrentlyevaluatingfordirectcommercialization.Although
thenutrientcompositionoftheseFLAVRSAVRtomatolineswassimilartothenutrientcompositionof
theparentalvarietiesandiswithintherangeofvaluesfortomatoesreportedintheliterature,mostofthe
nutrientcompositiondatawasbasedontheanalysisofsinglesamples(duetoalimitedamountof
availabletomatofruits).
However,inasubsequentsubmission,CalgeneincludedadditionaldataonthetwoFLAVRSAVRtomato
lines(CR3613andCR3623)thatthefirmcurrentlyisevaluatingforcommercialization.Asdiscussed
above,CalgenecomparedthenutrientcompositionofthesetwoFLAVRSAVRtomatolinestothenutrient
compositionoftheparentalvariety,andconductedthecomparativestudyoveraperiodoftime
correspondingtotheexpectedshelflifeoffreshmarkettomatoes.Webelievethatthedesignofthese
experimentsaddressesthespecificcommentsconcerningthetiming(relativetoharvest)ofCalgene's
analyticalmeasurements,andthereforearerelevanttoprocessingaswellasfreshmarketFLAVRSAVR
tomatoes.Further,wefindthatthesedatarefutethecomment'sassertionthatFLAVRSAVRtomatoes
mightexhibitadifferentnutritionalprofilethantomatoesderivedfromtheparentalvarietybecause,as
discussedabove,thelevelsofvitaminAandvitaminCinFLAVRSAVRtomatoesarecomparabletothe
levelsofthesesamenutrientsincontrol(parental)tomatoes.
Withrespecttothecomment'sassertionthatanFDAofficialhadpreviouslyindicated(nearly20years
ago)thattheagencywouldconsidera20%differenceinnutritionalvaluetobeasignificantchangeinthe
nutritionalvalueoffoodderivedfromacropplant,wenotethatalthoughthisconceptwasthesubjectof
informaldiscussion,FDAultimatelydidnotincorporateadefinitionof"significantchange"intothe
agency'sregulationsregardingtheGRASstatusofsubstancesofnaturalbiologicalorigin(21CFR170.30(f)
(2)).Wealsonotethattheconcentrationofprincipalnutrientsinfoodsderivedfromexistingplant
varietiesfrequentlyvariesbymorethan20%.Becausetheagency'sregulationsestablishnolimitsonthe
variabilityinthelevelsofnutrients,andinlightoftheknownvariabilityinnutrientlevelsinexistingplant
varieties,webelievethereisnoreasontorequestfromCalgeneastatisticalevaluationtodemonstrate
compliancewitha20%limitonthedifferencebetweenthenutrientcompositionofFLAVRSAVR
tomatoesandtheparentalvariety.
Insummary,wefindthatCalgenehasadequatelyaddressedwhethernutrientsinFLAVRSAVRtomatoes
arecomparabletonutrientsintheparentalvariety.

TheSafetyofFLAVRSAVRTomatoes
OnecommentmaintainedthatthesafetyassessmentofanewplantvarietysuchasFLAVRSAVR
tomatoesshouldnotbelefttothedeveloper.Thecitizenpetitionexpressedconcernthatfoodssuchas
FLAVRSAVRtomatoeswillbeallowedtoentercommercewithoutpremarketsafetytesting.
Asnotedinthe1992policystatement,developersofnewfoodshaveanobligationundertheacttoensure
thatthefoodstheyoffertoconsumersaresafeandincompliancewithallrequirementsoftheact(57FR
22984at22985),andultimately,itisthefoodproducerwhoisresponsibleforfoodsafety(57FR22984at
22991).Therefore,ifpremarkettestingisneededtoestablishthesafetyofafood,thefoodproducerdoes
havetheresponsibilitytoperformtheappropriatetesting.
Moreover,wenotethatCalgenerequestedconsultationwithFDAconcerningtheirsafetyassessment.We
evaluatedthesafetyassessmentconductedbyCalgeneandreachedtheconclusionthatFLAVRSAVR
tomatoeshavenotbeensignificantlyalteredwithinthemeaningof21CFR170.30(f)(2),whencompared
tovarietiesoftomatoeswithahistoryofsafeuse.Moreover,webelievethatthetestingconductedby
Calgeneisconsistentwithguidancesetoutinthe1992policystatementconcerningthesafetytestingof
foodsderivedfromnewplantvarieties.
OnecommentarguedthatCalgene'sdataonlevelsofthenaturallyoccurringplanttoxicanttomatine
providedrelativelylittlestatisticalpower.Becausetomatinelevelsdecreaseastomatoesripen,Calgene
couldnotdetecttomatinein97%ofthesamplesofripeFLAVRSAVRtomatoestested.Moreover,
tomatinelevelsinthe3%offruitsthatcontaineddetectablelevelswerecomparabletotomatinelevelsin
samplesofripecommercialtomatovarieties.Inlightofthesefindings,webelievethatastatistical
analysisofCalgene'sdataontomatinelevelsisnotessentialtothesafetyassessmentofFLAVRSAVR
tomatoes.

EconomicHarmResultingfromDecreasedConsumptionofTomatoes

ThecitizenpetitionmaintainedthatthereisasubstantialriskofconsumerconfusionbecausecurrentFDA
regulationswouldnotdistinguishtomatoesdevelopedusingtraditionalbreedingtechniquesfromnovel
foodssuchasFLAVRSAVRtomatoesandthatconsumersthereforemaychoosetoreducetheirpurchase
oftomatoes.Thecitizenpetitionarguedthatfarmerswhoderiveasignificantpartoftheirincomefrom
theproductionoftomatoesdevelopedusingtraditionalbreedingwillsufferseriouseconomicinjuryiftheir
productsareconfusedwithgeneticallyengineeredproductssuchasFLAVRSAVRtomatoes.
Thecitizenpetitiondidnotprovidedataorotherinformationtosupporttheassertionthat,intheabsence
oflabelingdistinguishinggeneticallyengineeredtomatoesfromtomatoesdevelopedusingtraditional
breeding,consumerconfusionwillresultinreducedpurchaseoftomatoesandcausefarmerstosuffer
seriouseconomicinjury.Inaddition,asdiscussedabove,wehavereviewedtheinformationsubmittedby
CalgeneconcerningFLAVRSAVRtomatoesinlightof(1)thestatutoryprovisionsforlabelingand
existingagencyregulationsandpolicyimplementingthosestatutoryprovisionsand(2)theopinionsofthe
agencyscientistswhoevaluatedtheinformationsubmittedbyCalgene.WehaveconcludedthatFLAVR
SAVRtomatoesdonotrequirespeciallabelingtocomplywiththeact.TheauthorityprovidedtoFDAby
theactdoesnotpermitFDAtopreventacompanyfromlegallymarketingaproductthatisincompliance
withtheactbecauseofpotentialeconomicconsequencesforcompetitors.
Moreover,asdiscussedabove,wenotethatCalgenehasannounceditsintentiontoprovidevoluntarilyto
consumerslabelingthatexplainsthatFLAVRSAVRtomatoesweredevelopedusinggeneticengineering
techniques.Thus,inthisparticularcase,therewouldappeartobenoriskthattheallegedeconomic
consequenceswouldoccur.

CONCLUSIONS
1. WeconsideredtheinformationprovidedbyCalgeneaswellastheinformationprovidedinthe
commentsandotherrelevantmaterial.Webelievethatthisinformationsupportsaconclusionthat
FLAVRSAVRtomatoeshavenotbeensignificantlyalteredwithinthemeaningof21CFR170.30(f)
(2),whencomparedtovarietiesoftomatoeswithahistoryofsafeuse.Inotherwords,theFLAVR
SAVRtomatoisassafeasothercommonlyconsumedtomatoes.
2. WerecommendthatOPAprovideCalgenewiththeopinion,thatFLAVRSAVRtomatoeshavenot
beensignificantlyalteredwithinthemeaningof21CFR170.30(f)(2),whencomparedtovarietiesof
tomatoeswithahistoryofsafeuse,byletterfromOPAratherthanasanadvisoryopinionunder21
CFR10.85.
3. WehaveconcludedthattheconsultationbetweenCalgeneandFDA,andanyletterorother
communicationfromFDAthatmayresult,isnotanagencyactionunderNEPA.
4. WebelievethatthecorrectcommonorusualnamefortheFLAVRSAVRtomatois"tomato",
becausetheFLAVRSAVRtomatoisnotsignificantlydifferentfromtherangeofcommercial
varietiesreferredtobythatname.Wealsohavedeterminedthatthereisnosafetyorusageconcern
towhichconsumersofFLAVRSAVRtomatoesmustbealertedbyspeciallabeling.
5. WebelievethatthesafetyassessmentconductedbyCalgeneisconsistentwiththeapplicable
provisionsoftheact,asreflectedinFDA'sguidancetoindustry,setoutinthe1992policystatement,
onscientificconsiderationsforevaluatingfoodsderivedfromnewplantvarieties.

LindaS.Kahl,Ph.D.

ThemeetingwasheldonApril68,1994,inHerndon,VA.

Theagencydidnotrecommendinthe1992policystatementthattoxicologicaltestingbeconducted
routinelyinpartbecauseitiswellrecognizedthatconventionalfeedingstudiesonwholefoodshave
limitedsensitivityandaresubjecttoconfoundingfactorsunrelatedtotestarticletoxicity,suchas
nutritionalorphysiologiceffects.
3

CalgenesubmittedinformationtotwoDocketNumbers:DocketNo.90A0416(requestforanadvisory

opinionontheuseofthekanrgene)andDocketNo.91A0330(requestforanadvisoryopiniononFLAVR
SAVRtomatoes).WehaveaddedtoDocketNo.91A0330literaturereferencesthatarecitedinthis
documentbutthatoriginallyweresubmittedbyCalgenetoDocketNo.90A0416.
Calgene'soriginalsubmissiontoDocketNo.91A0330hasbeenonpublicdisplayatDocketsManagement
BranchsinceAugust12,1991.Sincethattime,Calgenehassentanadditionalcopyofdatasubmittedfor
FDAevaluationdirectlytoDocketsManagementBranchforpublicdisplay.Aninformationalcopyof
Calgene'sPetitiontotheU.S.DepartmentofAgricultureAnimalandPlantHealthInspectionService(USDA
APHIS)requestingadeterminationoftheregulatorystatusofFLAVRSAVRtomatoesunder7CFR340
hasbeenpubliclyavailableatUSDAAPHISsinceJuly,1992andwasaddedtoDocketNo.91A0330in
December,1993.AcopyofCalgene'sletterdatedSeptember17,1992,requestingthatFDAreview
Calgene'svoluntarylabelingforFLAVRSAVRtomatoes,wasaddedtoDocketNo.91A0330inFebruary,
1993.
4

Opinesaresubstancesthatareexcretedbytumorcellsandserveasanenergysourcefortheinfecting
Agrobacterium.
5

SuchAgrobacteriumstrainsarecalled"disarmed"becausetheydonotcausecrowngalls.

CalgeneprovideddataonnutrientsotherthanvitaminAandvitaminC,andCFSANscientistscommented
onsuchadditionaldata.However,thismemorandumfocusesonvitaminAandvitaminCbecause
tomatoescontributesignificantamountsofthesevitaminstotheU.S.diet.Intheirdiscussions,theFood
AdvisoryCommittee(Ref.1)notedthatsomesubstancesthatoccurnaturallyintomatoesandcurrently
areuncharacterizedmaybenutritionallyimportant.Wenotethat,asknowledgeconcerningtheidentity
andlevelofimportantnutrientsinfoodssuchastomatoeschanges,wewouldexpectdevelopersto
conductappropriatetestingaccordingly.
7

Theamountofhomogenizedtomatoesadministeredtotheratsineachstudywasequivalenttoahuman
consumptionofapproximately10largeor40small(e.g.,plum)tomatoesperday.Theuseoftomatoes
thatwerelyophilizedandreconstitutedto50%oftheoriginalvolumedoubledthedoseoftomatoes(i.e.,
approximately20largeor80smalltomatoes)comparedtothedosethatcouldbeachievedusing
homogenizedtomatoes.
8

Thesymbol""indicatesthatthetreatmentgroupwasnotincludedinthatstudy.

AsreportedbyIRDC.

10

AsreportedbyPWGfollowingreviewofcodedmicroscopicslides.

11

InStudy3,Calgenereportedthegeographicsource(DixonorIndio)oftomatoesforeachgroup.

12

Geographicsource:Dixon.

13

Geographicsource:Dixon.

14

Geographicsource:Indio.

15

Theapprovalofafoodadditivepetition,ortheaffirmationofafoodingredientasgenerallyrecognized
assafe,unlikeaconsultationwithafoodproduceronthestatusofafoodproductorfoodsafetyissues,is
anagencyactionthatrequiresenvironmentalconsiderationunderNEPA.Inthenoticeannouncing
Calgene'srequestforanadvisoryopiniononthestatusofFLAVRSAVRtomatoes,FDAsummarized
Calgene'sargumentsinfavorofthefirm'sclaimthatFLAVRSAVRtomatoesarefoodthatissubjecttoa
categoricalexclusion(57FR22772at22773).Atthattime,FDAstatedthatthedecisionastowhether
CalgenemustfileanenvironmentalassessmentmightdependupontheregulatorystatusofFLAVRSAVR
tomatoesand,thus,theagencydeferredastatementofitspositionuntiltheagencyrespondedto
Calgene'srequest.Because,asaresultoftheconsultationbetweenCalgeneandFDA,theagencyisnot
requestingthatCalgenefileafoodadditivepetitionorGRASaffirmationpetitionforFLAVRSAVR
tomatoes,thereisnofurtherneedtoconsiderwhetherFLAVRSAVRtomatoeswouldwarranta
categoricalexclusion.
16

OurreasonsforbelievingthatCalgene'srequestforanadvisoryopiniononthestatusofFLAVRSAVR
tomatoesisconsistentwith21CFR10.85areasfollows.First,theseregulationsdonotrequirethata
requestbedeniediftherequestcoversaparticularproduct.Webelievethat,atthetimeCalgenefiledits
request,anadvisoryopinionunder21CFR10.85wasanappropriatemechanismtoobtainanagency
opiniononthestatusofawholefoodsuchasFLAVRSAVRtomatoesbecause,intheabsenceofan
announcedFDApolicyonthestatusoffoodsderivedfromnewplantvarieties,FLAVRSAVRtomatoes

raisedpolicyissuesofbroadapplicability.
Second,wedisagreethatCalgene'srequestforanadvisoryopinionviolated21CFR10.85(a)(2)(i)because
thesafetyofthekanrgeneforuseasaselectablemarkeringeneticallyengineeredwholefoodswasstill
underreview.PriortotherequestforanadvisoryopinionconcerningthestatusofFLAVRSAVR
tomatoes,Calgenerequestedanadvisoryopinionconcerningwhetherthekanrgenecanbeusedinthe
productionofgeneticallyengineeredtomato,cotton,andoilseedrapeplantsintendedforhumanfoodand
animalfeeduse.Thesafetyofthekanrgeneforuseasaselectablemarkeringeneticallyengineered
wholefoodswasthereforealreadyunderconsiderationandweseenoreasontobelievethattheagency
wouldhavedisregardedCalgene'srequestforanadvisoryopiniononthekanrgenewhenrespondingto
Calgene'srequestforanadvisoryopinionconcerningFLAVRSAVRtomatoes.
Third,theregulationsin21CFR10.85donotprohibitanyinterestedpersonfrommakingarequestforan
advisoryopinion,nordotheyprohibitFDAfromrespondingtothatrequestifFDAconsidersaresponseto
beinthepublicinterest.Webelievethat,becauseoftheissuesraisedbyCalgene'srequest,itwasinthe
publicinterestforFDAtoconsiderCalgene'srequest.ThenoticeannouncingCalgene'srequestprovided
thepublicanopportunitytocommentonthoseissues.
17

WearenotawareofanypublishedstudiesdemonstratingthesafetyofanyspecificRNAthatispresent
inthecellsofanyfoodcrop,norwouldweexpectsuchpublicationbecause,asdiscussedabove,nucleic
acidsarepresentinthecellsofeveryplantusedforfood,areefficientlydigested,anddonotthemselves
raiseasafetyconcernasacomponentoffood.
18

Furthermore,APH(3')IIsatisfiesthedefinitionof"processingaid"in21CFR101.100(a)(3)(ii)(c),andwill
beregulatedassuchfollowingtheagency'sdecisiononFAP3A4364.Asthecommentacknowledges,FDA's
labelingregulationsexemptprocessingaidslikeAPH(3')IIfromthelabelingrequirementsofsection403(i)
(2)oftheact.Thus,evenifAPH(3')IIwereproperlyconsideredaningredient,itspresenceinafoodwould
notberequiredtobedisclosedinthefood'slabeling.

REFERENCES
NOTE:Becauseoftheirsize,References#1and#2arenotattachedtothismemorandum.References
markedwithasterisks(***)areavailableinthepublishedliterature,aswellasintheadministrativefile
oftheFLAVRSAVRtomato,andalsoarenotattachedtothismemorandum.
1. TranscriptofthemeetingofFDA'sFoodAdvisoryCommittee,Herndon,VA,April68,1994.
2. BackgroundInformation:FDA'sConsultationwithCalgene,Inc.,ConcerningFLAVRSAVRTomatoes.
3. LetterdatedJune3,1993,fromDonaldL.EmlayofCalgene,Inc.,toDocketsManagementBranch.
4. MemorandumofTelephoneConversationsbetweenLindaKahlofFDAandDonEmlayofCalgene,
Inc.,onOctober6and7,1993.
5. Forexample,seethefollowing:
A. ***Senti,F.R.andR.L.Rizek,"NutrientLevelsinHorticulturalCrops,"Hort.Science10:243
246,1975.
B. ***Davies,J.N.andG.E.Hobson,"TheConstituentsofTomatoFruitTheInfluenceof
Environment,Nutrition,andGenotype,"CRCCrit.Rev.inFoodSci.andNutr.15:205280,1981.
6. Forexample,seethefollowing:
A. ***Fillatti,J.J.,etal.,"EfficientTransferofaGlyphosphateToleranceGeneintoTomatousinga
BinaryAgrobacteriumtumefaciensvector,"Bio/Technology5:726730,1987.
B. ***McBride,K.E.andK.R.Summerfelt,"ImprovedBinaryVectorsforAgrobacteriummediated
PlantTransformation.PlantMol.Biol.14:269276,1990.
7. MemorandumofMeetingbetweenrepresentativesofFDAandCalgene,Inc.,onMay15,1991.
8. MemorandumofMeetingbetweenrepresentativesofFDAandCalgene,Inc.,onNovember19,1991.
9. ***Hoskins,L.C.,"HostandmicrobialDNAintheGutLumen,"JournalofInfectiousDiseases,

137:694698,1978.
10. LetterdatedSeptember17,1992fromKeithRedenbaughofCalgene,Inc.,toJamesH.Maryanskiof
FDA.
11. ***Spiher,A.T."FDARegulations:ANewDevelopmentinAgriculture,"in"TheEffectofFDA
Regulations(GRAS)onPlantBreedingandProcessing,"CSSASpecialPub.No.5,Hanson,C.H.etal.,
editors,CropSci.Soc.ofAmerica,Madison,WI,pp.15,1974.

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