TheSucrose/D-Fructose/D-Glucosetestkitissuitablefor themeasurementandanalysisofsucrose,D-glucoseandD-fructoseinplantandfoodproducts.
Grapeandwineanalysis:Oenologiststoexploitadvancedtestkits.
Charnock,S.C.&McCleary,B.V.(2005).RevuedesEnology,117,1-5.
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Itiswithoutdoubtthattestingplaysapivotalrolethroughoutthewholeofthevinificationprocess.Toproducethebestposs
IBLequalitywineandtominimiseprocessproblemssuchas“stuck”fermentationortroublesomeinfections,itisnowrecognisedthatifpossibletestingshouldbeginpriortoharvestingofthegrapesandcontinuethroughtobottling.Tr
ADItionalmethodsofwineanalysisareoftenexpensive,timeconsuming,requireeitherelaborateequipmentorspecialistexpertiseandfrequentlylackaccuracy.However,enzymaticbio-analysisenablestheaccuratemeasurementofthevastmajorityofanalytesofinteresttothewinemaker,usingjustonepieceofapparatus,thespectrophotometer(
seepreviousissueNo.116foradetailedtechnicalreview).Grapejuiceandwineareamenabletoenzymatictestingasbeingliquidstheyarehomogenous,easytomanipulate,andcangenerallybeanalysedwithoutanysamplepreparation.
Megazyme“advanced”winetestkitsgeneralcharacteristicsandvalidation.
Charnock,S.J.,McCleary,B.V.,Daverede,C.&Gallant,P.(2006).ReveuedesOenologues,120,1-5.
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ManyoftheenzymatictestkitsareofficialmethodsofprestigiousorganisationssuchastheAssociationofOfficialAnalyticalChemicals(AOAC)andtheAmericanAssociationofCerealChemists(AACC)inresponsetotheinterestfromoenologists.Megazymedecidedtouseitslonghistoryofenzymaticbio-analysistomakeasignificantcontributiontothewineindustry,bythedevelopmentofarangeofadvancedenzymatictestkits.Thistaskhasnowbeensuccessfullycompletedthroughthestrategicandcomprehensiveprocessofidentifyinglimitationsofexistingenzymaticbio-analysistestkitswheretheyoccurred,andthenusingadvancedtechniques,suchasmolecular
BIOLOGy(
photo1),torapidlyovercomethem.Noveltestkitshavealsobeendevelopedforanalytesofemerginginteresttotheoenologist,suchasyeastavailablenitrogen(
YAN;seepages2-3ofissue117article),orwherepreviouslyenzymesweresimplyeithernotavailable,orweretooexpensivetoemploy,suchasforD-mannitolanalysis.
EvaluationofTypeofProcessonFunctionalPropertiesandonNutritionalandAnti-nutritionalCompositionofYams(Dioscoreacayenensis-rotundata).
Kouassi,N.K.,Nindjin,C.,Tetchi,A.F.&Amani,G.N.(2010).JournalofFoodTechnology,8(4),191-199.
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Thenutritional,anti-nutritionalcompositionandfunctionalpropertiesofyamfloursobtainedfromdifferentprocesseswithfourvarietiesofDioscoreacayenensis-rotundataextensivelyconsumedinCoted’Ivoirewereevaluated.Theresultsshowedthatnutritionalandanti-nutritionalcompositionofyamsareloweranddecreasegreatlyduringboilingthanbaking.Thelowervaluesofleastgelatinizationconcentrationobtainedwithflourofcookedyamthanrawyamflourisduetothelossofamylosewhichisassociatedwithstarchgranulesgelatinizationduringthecooking.Thisgelatinizationischaracterizedbygranulestarchofhighsizeandaheterogeneousdistribution.Thegreatsolubilityandtheweakcapacityofswellingduringheatingofflourofboiledyamareduetothefactthatthegelatinizationofstarchhadbeenhigherintheboiling.Thisgreatgelatinizationinboiledyamisassociatedtothehighlevelofglucoserateduringdigestionandcouldinducemetabolicdisorders.
SubmergedcultureprocessforbiomassandexopolysaccharideproductionbyAntarcticyeast:someengineeringconsiderations.
Vlaev,S.,Rusinova-Videva,S.,Pavlova,K.,Kuncheva,M.,Panchev,I.&Dobreva,S.(2013).AppliedMicrobiologyandBiotechnology,97(12),5303-5313.
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Productionofbiomassandextracellularpolysaccharide(EPS)frompsychrophilicSporobolomycessalmonicolorAL1inastirredbioreactorwasstudied.Theaspectsofproductiontechnical-scaleparameters,namely,bioreactorflowfield,biomassandEPSproductionrates,oxygenmasstransferperinputpower,aswellasimportantproductproperties,suchasrheologyandstabilityofEPSmixtures,wereconsidered.Thebioprocesswasfoundtoproceedinnon-Newtonianflowwithconsistencycoefficientrisingtypicallyto0.03Pa.snandflowindexdecliningto0.7.Flowmodelingwascarriedoutandshowedgoodhomogenizationforsubstratedeliveryatagitationratesexceeding400rpm.Agitationrateslowerthan400rpmwereconsideredcounterproductiveduetoflowfieldnon-uniformity.Thecelldensityreached5g/landEPSproductionyieldreached5.5g/latproductionrate0.057gEPS/lperhour(0.01gEPS/gbiomassperhour).Oxygenuptakerateandoxygentransferratewereintherangeof0.5–1.7mmolO2/lperhourand2–4.7mmolO2/lperhour,respectively.ThemasstransfercoefficientatreactionconditionswasfoundtobeintherangeKLa~0.004−0.01s-1.Thebioprocessbiologicalperformancewashigheratmoderateagitationspeedandrevealedbiomassdiminutionandcellinactivationbyincreasingimpellerrevolutionsandshearrate.TheproductEPSwasfoundtointroduceshear-thinningbehaviorinwatersolutionswithapparentviscosityofupto30mPa.sandtostabilize1–2%oil-in-wateremulsionsimprovingtheirlipophilicproperties.TheemulsiondispersionindexwasfoundtobecomparablewiththeoneofArlacel165,theemulsifierusedincosmetic.Thelong-termperformanceofthecomplexcreammixturesoftheglucomannanpreparedincommercialformatwasfoundpromisingforfurtherapplication.
Near‐infraredreflectancecalibrationsfordeterminingsucrosecontentinsoybeanbreedingusingartificialreferencesamples.
Sato,T.,Zahlner,V.,Berghofer,E.,Lošák,T.&Vollmann,J.(2012).PlantBreeding,131(4),531-534.
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Infood-gradesoybeansforhumannutrition,sucrosecontentisacharacterofinterestapartfromprotein,becausesucroseaffectsflavourandothersoy-foodproperties.Asdeterminationofsucrosecontentthroughseparativeorenzymaticmethodsisnotsuitableforanalysinglargenumbersofsamples,theobjectiveofthisresearchwastodevelopnear-infraredreflectancespectroscopy(NIRS)calibrationsforrapidscreeningofbreedinglinesforsucrose.Conventionalsoybeansampleshaveasucrosecontentof30–60g/kg,whereashigh-sucrosereferencesamplesarelacking.Therefore,bothnaturalandartificial(additionalsucroseadded)referencesampleswithasucrosecontentofupto140g/kgwerecombined,andaNIRScalibrationmodelwasdeveloped(cross-validationR2=0.969)forsucrose.Utilizingwavelengthrangeswithhomogeneousperformanceinbothnaturalandartificialsamples,apredictionmodelwasobtained,whichsuggestsastableperformanceofthecalibrationoverawiderangeofsucroseconcentrations.Thus,identificationofhigh-sucrosesoybeanintrogressionsshouldbepossibleinappropriatesegregatingpopulations.
AlteredseedoilandglucosinolatelevelsintransgenicplantsoverexpressingtheBrassicanapusShootmeristemlessgene.
Elhiti,M.,Yang,C.,Chan,A.,Durnin,D.C.,Belmonte,M.F.,Ayele,B.T.,TahirM.&Stasolla,C.(2012).JournalofExperimentalBotany,63(12),4447-4461.
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SHOOTMERISTEMLESS(STM)isahomeoboxgeneconservedamongplantspecieswhichisrequiredfortheformationandmaintenanceoftheshootmeristembysuppressingdifferentiationandmaintaininganundeterminedcellfatewithintheapicalpole.Toassessfurthertheroleofthisgeneduringseedstorageaccumulation,transgenicBrassicanapus(Bn)plantsoverexpressingordown-regulatingBnSTMunderthecontrolofthe35Spromoterweregenerated.OverexpressionofBnSTMincreasedseedoilcontentwithoutaffectingtheproteinandsucroselevel.Thesechangeswereaccompaniedbytheinductionofgenesencodingseveraltranscriptionfactorspromotingfattyacid(FA)synthesis:LEAFYCOTYLEDON1(BnLEC1),BnLEC2,andWRINKLE1(BnWRI1).Inaddition,expressionofkeyrepresentativeenzymesinvolvedinsucrosemetabolism,glycolysis,andFAbiosynthesiswasup-regulatedindevelopingseedsectopicallyexpressingBnSTM.ThesedistinctiveexpressionpatternssupporttheviewofanincreasedcarbonfluxtotheFAbiosyntheticpathwayindevelopingtransformedseeds.TheoverexpressionofBnSTMalsoresultedinadesirablereductionofseedglucosinolate(GLS)levelsascribedtoatranscriptionalrepressionofkeyenzymesparticipatingintheGLSbiosyntheticpathway,andpossiblytothedifferentialutilizationofcommonprecursorsforGLSandindole-3-aceticacidsynthesis.NochangesinoilandGLSlevelswereobservedinlinesdown-regulatingBnSTM.Takentogether,thesefindingsprovideevidenceforanovelfunctionforBnSTMinpromotingdesirablechangesinseedoilandGLSlevelswhenoverexpressedinB.napusplants,anddemonstratethatthisgenecanbeusedasatargetforgeneticimprovementofoilseedspecies.
Engineeringcyanobacteriatosynthesizeandexporthydrophilicproducts.
Niederholtmeyer,H.,Wolfstädter,B.T.,Savage,D.F.,Silver,P.A.&Way,J.C.(2010).AppliedandEnvironmentalMicrobiology,76(11),3462-3466.
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MetabolicengineeringofcyanobacteriahastheadvantagethatsunlightandCO2arethesolesourceofenergyandcarbonfortheseorganisms.However,asphotoautotrophs,cyanobacteriagenerallylacktransporterstomovehydrophilicprimarymetabolitesacrossmembranes.Toaddresswhethercyanobacteriacouldbeengineeredtoproduceandsecreteorganicprimarymetabolites,SynechococcuselongatusPCC7942wasengineeredtoexpressgenesencodinganinvertaseandaglucosefacilitator,whichmediatedsecretionofglucoseandfructose.Similarly,expressionoflactatedehydrogenase-andlactatetransporter-encodinggenesallowedlactateaccumulationintheextracellularmedium.Expressionoftherelevanttransporterwasessentialforsecretion.Productionofthesemoleculeswasfurtherimprovedbyexpressionofadditionalheterologousenzymes.SugarssecretedbytheengineeredcyanobacteriacouldbeusedtosupportEscherichiacoligrowthintheabsenceofadditionalnutrientsources.Theseresultsindicatethatcyanobacteriacanbeengineeredtoproduceandsecretehigh-valuehydrophilicproducts.
Variationandcorrelationofpropertiesindifferentgradesofmaplesyrup.
Singh,A.S.,Jones,A.M.P.&Saxena,P.K.(2014).PlantFoodsforHumanNutrition,69(1),50-56.
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ThirtyfivecommercialmaplesyrupsfromtwelveproducersinSouthernOntariowereevaluatedforpropertiesincludinglighttransmittance,autofluorescence,density,pH,totalsolublesolids(TSS),glucoseandfructosecontent,totalphenolcontent,antioxidantpotentialandmineralcontent(Mg,Mn,P,Zn,Ca,K,FeandPb).Ahighdegreeofvariabilitywasfoundinmanycharacteristics,oftenexceedinganorderofmagnitude.Syrupswerecategorizedbasedonlighttransmissionat560nmintoamber(12),dark(13)andverydark(10)usingInternationalMapleSyrupInstitute(IMSI)guidelines.Nostatisticaldifferenceswerefoundamonggradesofsyrupfordensity,pH,TSS,glucose,fructose,totalreducingsugars,glucose:fructoseratio,magnesium,manganeseorpotassium.Darkersyrupsshowedsignificantlyhigherautofluorescence,totalphenolcontent,antioxidantpotential,phosphorous,calciumandtotalmineralcontent.Significantnegativecorrelationsofpercenttransmissionwithtotalphenolcontent,antioxidantpotentialandtotalmineralcontentarereported.Significantpositivecorrelationsamongtotalphenolcontent,antioxidantpotentialandtotalmineralcontentarealsodescribed.Theresultsfromthisstudysuggestthatdarkersyrupstendtocontainmorebeneficialtraitsandmaybeappliedindevelopingfunctionalfoodsandvalueaddedproducts.
Spontaneouspostharvestfermentationofaçai(Euterpeoleracea)fruit.
Aguiar,F.,Menezes,V.&Rogez,H.(2013).PostharvestBiologyandTechnology,86,294-299.
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Açai(Euterpeoleracea)fruit(EOF)arewidelycommercializedintheBrazilianAmazon.Thesefruitcontainahighbacterialloadandaretransportedonboardsstowedinsideoroutsidetheholdsofsmallboats.Inthiscontext,postharvestparameterswereassessedunderconditionsthatsimulatedthesetwomethodsofEOFtransport:stowageinclosedpolystyreneboxes,simulatingtheinsideofcargoholds,i.e.,transportinaclosedsystem;andopenbaskets,simulatingtransportinanopenenvironment,i.e.,transportintheproworbowoftheboat.EOFsufferedspontaneousfermentationofalcoholic,acetic,andlactictypesintheclosedsystem,whichisthemostcommontypeoftransportationofthisfruit.Intheclosedsystem,therewasapredominanceoflacticacidbacteriaoveraceticacidbacteria,with82%and95%oftheinitialcontentofD-glucoseandD-fructosebeingconsumed,respectively,after27hofexperiment.Theweightlossreached1.7%andtherewasalogarithmicdecreaseofthemajorphenoliccompoundsofthefruitintheclosedsystem,withlossesof78%ofcyanidin-3-rutinoside,88%ofcyanidin-3-glucoside,78%ofhomorientin,and72%oforientinafter27h,whichwashigherthanintheopensystem(58%,66%,73%and62%,respectively).AnalysesonEOFstowedinaclosedsystemindicatedthattherespiratoryratewascharacteristicofanon-climactericfruit,i.e.,itshowedalogarithmicdecayintheproductionofCO2(R2=0.995;P<0.05).=""thus,=""transport=""in=""a=""closed=""system=""results=""in=""more=""drastic=""nutritional=""and=""functional=""changes=""on=""eof=""than=""when=""transport=""is=""carried=""out=""in=""an=""open=""system,=""suggesting=""that=""transportation=""in=""continuous=""aerobic=""conditions=""and=""a=""short=""period=""of=""time=""between=""picking=""and=""processing=""are=""preferable.="">
PartialStudyofYamTuber(Dioscoreaspp.)PartsduringtheGrowthPeriod.
ClaverDegbeu,K.,N"dri,Y.D.,FabriceTetchi,A.,Nindjin,C.,N"guessanAmani,G.&Bakayoko,A.(2013).AdvanceJournalofFoodScience&Technology,5(9),1120-1131.
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Growinggradientintolongitudinalaxisofyamtuberwasstudythroughstarchpropertiesandnutritionalcompositionoftuberparts(proximal,mediananddistal)duringthetuberization.TwovarietiesofthecomplexD.cayenensis-rotundata(kangbaandkponan)wereused.Clarity,swellingandsolubility,flowbehaviorandsyneresisofstarchandproteincontent,alcohol-solublesugarsoftheflourwerestudies.Theashassessedbymicroanalysis.Starchclarityoftuberpartsincreasedduringthegrowthperiod.Theproximal(56±2.06%)andmedian(54.5±1.09%)partsexhibitedhighclaritythanthedistal(48.2±2.56%)one.Atearlierstageoftuberization,theviscosityratiooftuberparts(var.Kponan)wasweak.Thisindicatesthepossibilitytouseitasthickeningagentinshearingsauce.Waterlossdecreasedduringthegrowthperiodforthethreetuberparts.Itwasthesamebehaviorfortheswellingpower.Concerningthenutritionalcompositionoftheflour,amountofproteindidnotinfluencedbythedegreeoftubermaturity.Alcohol-solublesugarswerehigheratearlierstageoftuberizationthanatmaturityofthetuber.Mineralcontentwasappreciableatmaturitythanatbeginningofthetuberization.Theextentmineralaccordingtotheiramountwas:K,P,MgandS.Starchpropertiessuchasclarity,syneresisandswellingwereimprovedatmaturityofthetuber.Itwasmorefortheproximalandmedianpartsthanthedistalone.Yamtubercontentaappreciableamountofproteinandmineralatmaturity.
GalanthamineproductionbyLeucojumaestivumL.shootcultureinamodifiedbubblecolumnbioreactorwithinternalsections.
Georgiev,V.,Ivanov,I.,Berkov,S.,Ilieva,M.,Georgiev,M.,Gocheva,T.&Pavlov,A.(2012).EngineeringinLifeSciences,12(5),534-543.
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Shootcultureofsummersnowflake(LeucojumaestivumL.)wassuccessfullycultivatedinanadvancedmodifiedglass-columnbioreactorwithinternalsectionsforproductionofAmaryllidaceaealkaloids.Thehighestamountsofdrybiomass(20.8g/L)andgalanthamine(1.7mg/L)wereachievedwhenshootswereculturedat22°Cand18L/(L•h)flowrateofinletair.Attheseconditions,theL.aestivumshootculturepossessedmixotrophic-typenutrition,synthesizingthehighestamountsofchlorophyll(0.24mg/gDW(dryweight)chlorophyllAand0.13mg/gDWchlorophyllB).Thealkaloidsextractofshootbiomassshowedhighacetylcholinesteraseinhibitoryactivity(IC50=4.6mg).Thegaschromatography–massspectrometry(GC/MS)profilingofbiosynthesizedalkaloidsrevealedthatgalanthamineandrelatedcompoundswerepresentedinhigherextracellularproportionswhilelycorineandhemanthamine-typecompoundshadhigherintracellularproportions.Thedevelopedmodifiedbubble-columnbioreactorwithinternalsectionsprovidedconditionsensuringthegrowthandgalanthamineproductionbyL.aestivumshootculture.
Sugaraccumulationinrootsoftwograpevarietieswithcontrastingresponsetowaterstress.
Rogiers,S.Y.,Holzapfel,B.P.&Smith,J.P.(2011).AnnalsofAppliedBiology,159(3),399-413.
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Rootsugaraccumulationwasstudiedintwograpevinevarietiescontrastingintolerancetowaterstress.Duringa10-daywaterwithholdingtreatment,thedrought-tolerantvariety,Grenache,sustainedlessnegativepredawnandmiddayleafwaterpotentialsaswellasrootwaterpotentialcomparedwiththesensitivevariety,Semillon.GrenachevinesalsomaintainedlowerstomatalconductanceandtranspirationthanSemillonvinesthroughoutthedryingperiod.Inbothvarietiestherewasaccumulationofsucroseintherootsandconcentrationswereinverselycorrelatedtoleafandrootwaterstatus.InbothGrenacheandSemillon,elevatedrootosmolalitywasassociatedwithdecreasedsoilmoistureindicatingthatsugaraccumulationmayplayaroleinosmoticprotection.Petiolexylemsapabscisicacid(ABA)concentrationsincreasedwithwaterdeficitinbothvarietiesandwerehighestforvineswiththemostnegativerootandpredawnleafwaterpotentials.Fur
Thermore,rootsucroseconcentrationswerepositivelycorrelatedwithleafxylemsapABAconcentrations,indicativeofintegrationbetweencarbohydratemetabolismandtheABAsignallingsystem.Similarrootsugaraccumulationpatternsbetweenthetwovarieties,however,demonstratethatotherfactorsarelikelyinfluencingtheabilityofthedrought-tolerantvarietytoremainhydrated.
CarbohydratereservestatusofMalbecgrapevinesafterseveralyearsofregulateddeficitirrigationandcroploadregulation.
Dayer,S.,Prieto,J.A.,Galat,E.&PerezPeña,J.(2013).AustralianJournalofGrapeandWineResearch,19(3),422-430.
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BackgroundandAims:Regulateddeficitirrigation(RDI)andbunchthinningaretwoviticulturalpracticesappliedworldwide.Thereislimitedknowledge,however,abouttheircombinedeffectsoncarbohydratedynamicsandaccumulation.Weevaluatedduringyear3and4oftheexperimenttheeffectof4consecutiveyearsofRDIandbunchthinningoncarbohydratestatus,vegetativeandreproductivevariables.MethodsandResults:From2006/07to2009/10,weimposedfourlevelsofwatersupply[100,60,38and25%ofreferenceevapotranspiration(ETo)]andtwocroploads(100and50%ofthebunches).Wecomparedshootlength,flowersperinflorescenceandyieldin2009/10.Wealsomeasuredpruningmassandtheconcentrationofnon-structuralcarbohydratesindormantwoodinthewintersof2009and2010.Starchconcentrationinthetrunkwasreducedbyseverewaterdeficit(25and38%ETo)andimprovedbybunchthinning.Pruningmass,shootlength,flowersperinflorescenceandyieldwereaffectedinvineswithwaterappliedat25and38%ofETo.Conclusions:Severewaterstressandhighcroploadreducedtrunkstarchconcentrationwithnointeractionbetweenbothfactors,whereastheconcentrationoftotalnon-structuralcarbohydratewasnotaffected.Vegetativegrowthandyieldwerereducedafter4yearsofseverewaterstress.SignificanceoftheStudy:Weprovideevidencethatstarchconcentrationandcarbonpartitioningcanbemanipulatedthroughcommonviticulturalpractices,suchasirrigationandcropload.
Stemcarbohydratedynamicsandexpressionofgenesinvolvedinfructanaccumulationandremobilizationduringgraingrowthinwheat(TriticumaestivumL.)genotypeswithcontrastingtolerancetowaterstress.
Yáñez,A.,Tapia,G.,Guerra,F.&delPozo,A.(2017).PloSone,12(5),e0177667.
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Thegeneticandphysiologicalmechanismsunderlyingtherelationshipbetweenwater-solublecarbohydrates(WSC)andwaterstresstolerancearescarcelyknown.ThisstudyaimedtoevaluatethemainWSCinstems,andtheexpressionofgenesinvolvedinfructanmetabolisminwheatgenotypesgrowinginaglasshousewithwaterstress(WS;50%fieldcapacityfromheading)andfullirrigation(FI;100%fieldcapacity).Eightwheatgenotypes(fivetolerantandthreesusceptibletowaterstress)wereevaluatedinitially(experiment1)andthetwomostcontrastinggenotypesintermsofWSCaccumulationwereevaluatedinasubsequentexperiment(experiment2).MaximumaccumulationofWSCoccurred10–20daysafteranthesis.UnderWS,thestress-tolerantgenotypeexhibitedhigherconcentrationsofWSC,glucose,fructoseandfructaninthestems,comparedtoFI.Inaddition,thestress-tolerantgenotypeexhibitedhigherup-regulationofthefructan1-fructosyltransferaseB(1-FFTB)andfructan1-exohydrolasew2(1-FEHw2)genes,whereasthesusceptiblecultivarpresentedanup-regulationofthefructan6-fructosyltransferase(6-SFT)andfructan1-exohydrolasew3(1-FEHw3)genes.OurresultsindicatedcleardifferencesinthepatternofWSCaccumulationandtheexpressionofgenesregulatingfructanmetabolismbetweenthetolerantandsusceptiblegenotypesunderWS.