电子记录稻飞虱取食和产卵行为及其在植物抗虫性研究中的应用_英文_

ElectronicMonitoringofFeedingandOvipositionBehaviorofRicePlanthoppersandItsApplicationinPlantResistanceStudy

HATTORIMakoto

(NationalInstituteofAgrobiologicalSciences,Tsukuba30528634,Japan;E2mail:hatto@affrc.go.jp)

电子记录稻飞虱取食和产卵行为及其在植物抗虫性研究中的应用

服部　诚　(日本国家农业生物资源研究所,日本筑波30528634;

E2mail:hatto@affrc.go.jp)

摘　要:采用AC型电子记录系统(EMS)对褐飞虱在非寄主植物稗草和具有抗性基因bph4的水稻品种Babawee上的取

食行为进行了比较研究。结果表明,褐飞虱口针到达稗草韧皮部以前停止取食,这揭示了在稗草非韧皮部组织中存在拒食剂。而在抗虫品种上,口针到达韧皮部以后取食行为终止。因此,可以清楚地识别两类具有不同抑制取食抗性因子的抗性植物。分析白背飞虱产卵行为的电子记录表明,在大多数情况下,就旺盛生长的水稻植株的下半部分而言,白背飞虱产卵器插入植物组织中但不产卵。这表明旺盛生长的植株能诱导白背飞虱的产卵行为,但由于某些拒避性因素使整个产卵过程不能完成。因此,这一系统可以有效地识别具有产卵抗性的植物上行为序列中受到破坏的步骤。关键词:褐飞虱;白背飞虱;电子记录系统;取食行为;产卵行为;抗虫性;稗草;水稻文献标识码:A文章编号:100127216(2003)增刊20031206中图分类号:Q6233;S433;S435.112+.3

Abstract:FeedingbehaviorsofNilaparvatalugensonanon2hostplant,thebarnyardgrassEchinochloacrus2gallivar.

oryzicola,andresistantriceBabawee(bph4)werecomparedbyusingAC2eletronicmonitoringsystem(EMS).Waveformdataob2

tainedbyEMSshowedthatN.lugensusuallystoppedfeedingactivitiesbeforestyletsreachthephloeminbarnyardgrasswhichre2vealedtocontainanantifeedantinthenon2phloem,whereasfeedingwasinterruptedafterstyletsreachedthephloeminresistantrice.Thus,EMSanalysisclearlydistinguishedtwotypesofresistantplantswithdifferentresistantfactorsagainstplanthopperfeeding.EMSanalysisofSogatellafurciferaovipositionbehavioronthelowerpartoftheleafsheathoffully2grownricedemonstratedthatinmostcasestheovipositorwasinsertedintotheplantbutpulledoutwithoutdepositinganyeggs.Itsuggestedthatovipositionbehavioritselfwasnormallyinducedonthefully2grownrice,buttheentireprocesswasnotcompletedduetosomedeterringfactors.Thus,thissystemisalsoeffectiveinidentifyingadisruptedstepinthebehavioralsequenceonaplantresistanttooviposition.

Keywords:Nilaparvatalugens;Sogatellafurcifera;electronicmonitoringsystem;feedingbehavior;ovipositionbehavior;in2sectresistantce;Echinochloacrus2galli;rice

1　Introduction

ACelectronicmonitoringsystem(AC2EMS)wasoriginallydevelopedtostudyfeedingbehaviorofaphids[14].Subsequently,DCsystemwasalsodevised[18]andimproved[22].TheEMShasbecomethemostaccurateanddirectmethodbywhichtheprobingactivitiesofpiercing2suckinginsectswithintheplanttissuecanbemonitored[1].Ithasbeenusedasatoolinthestudyofplantre2sistancetoaphidsandotherhomopteraninsects,toidentifythemechanismsofresistance,antixenosis,antibiosisandtoler2ance[9,17,24].Currently,thistechniquewasalsoappliedforana2lyzingovipositionbehaviorofthebrownplanthopper(BPH),

downunderseverepopulationpressure.Abreedingapproachbycombiningorpyramidingmajorresistantgenesisimportantforproducinghighlevelsofresistantvarieties[3].Forthispurpose,efficientmethodsforevaluatingvariouskindsofresistancewillberequired.

Inthisstudy,itwasconfirmedthattheEMSisausefultoolinelucidatingmechanismsunderlyingplantresistancetofeedingofpiercing2suckinginsectsandovipositionoflacerating2depositinginsects.Inthispaper,thecorrelationsbetweenvariouswave2formsandbehavioraleventsofBPHfeedingandovipositionwerefirstdescribedandthenwaveformoutputswerecomparedbe2tweenBPHandWBPH.Onthebasisofthewaveforminterpreta2tion,feedingbehaviorofBPHonanon2hostplant,whoseresis2tantfactorwasdeterminedchemically,wasrecordedbyusingthe

收稿日期:2003201221。

第一作者简介:服部　诚(19502),男,博士,室长。

…l,whichlaceratesplanttissuewithitsNilaparvatalugens(St)

saw2likeovipositoranddepositseggs[5].

BPHandthewhitebackedplanthopper(WBPH),Sogatella)arerecognizedasseriouspestsinAsia,in2ath　furcifera(Horv’

cludingJapanandChina.Sofar,ricevarietieswitheachofthreeresistantgenestoBPHhavebeenreleased,buthaveoftenbroken

32ChineseJournalofRiceScience,Vol.17,Suppl,2003

EMS,anditswaveformprofileswerecomparedwiththoseofaresistantvarietyofrice.Furthermore,behaviorofWBPHonaplantunsuitableforovipositionwasalsomeasuredtoidentifythestepinwhichovipositionsequencewasinterrupted.

eydewdropletsandcutstylets.Sequentialspotsofhoneydewdroplets(ca.25/h)collectedonafilterpaperduringsustainedIwaveform(over30min)alwaysshowedhighninhydrinandposi2tiveforsugar.Furthermore,cutstyletsnearlyalwaysexudeddropletsofphloemsapiftheywerecutduringsustainedIwave2form.TheIwaveformalwaysimmediatelyfollowedaseriesoftheAwaveform.ItisreportedthatduringtheAwaveformtheterminallocationsoffeedingtrackendedinphloem[11].Wave2formsobservedinBPHduringfeedingactivitiesstronglyresem2bledthoseinWBPH,althoughtheAwaveformcontinuedforashorterperiodanditsvoltagelevelwasrelativelyunstable(Fig.2).Therefore,theinterpretationofwaveforminBPHwillbeapplicablefortheWBPH’s.

2.2　DifferencebetweenfeedingbehaviorofBPHonanon2hostandaresistanthostplant

ThefeedingactivityofBPHwasrecordedonthebarnyardgrass,Echinochloacrus2gallivar.oryzicola,andonaresistantvarietyofrice,Babawee(bph4),byusingtheEMS[6].Thebarnyardgrassisregardedasamodelplantforanalyzingfeedinginterruptionprocess,becausethisspecieshasbeenproventocon2tain(E)2aconiticacid,anantifeedantforBPH[12]innon2phloemtissueofleafsheath[8].ComparisonofthefeedingpatternofBPHontworesistantplantsallowedcleardifferentiationofprobingprofiles.Onthebarnyardgrass,theplanthoppersperformedmanyprobes,butthemeandurationofaprobeperinsectwasshorterthanthatontheresistantandsusceptiblerice(Fig.3).Furthermore,thepercentageofprobesthatconsistedonlyofSwaveform(salivation)werehigheronthebarnyardgrassthanthatontheresistantrice,accountingformorethan50%oftotalprobes(Fig.4).Ontheotherhand,ontheresistantrice,theIwaveformwasobservedasfrequentlyasitwasonthesusceptiblerice.However,itsdurationontheresistantricewasmuchshort2erthanthatonthesusceptibleone(Fig.5).

2　ACelectronicmonitoringsystem(EMS)

TheAC2EMSrecordschangesinelectricalresistanceoftheinsect2plantinterface,whiletheDC2systemisabletorecordresis2tantchangesandelectromotiveforceoftheinsectandplant[1].However,forcollectionofdataonsalivationandphloeminges2tion,ACandDCsystemsareequallyuseful[16].Inthisstudy,ACvoltage(500Hz,0.5V)wasintroducedintotheplanthop2per2plantinterface.Whenawiredinsectpenetratesitsstyletsorovipositorintotheplant,thecircuitiscompletedandthenwave2formscharacterizedbyamplitude,frequencyandvoltagelevelarerecorded,dependingonspecificactivitiesoftheinsectduringfeedingandoviposition.Styletsandovipositoractivitiesincludingpenetratingandejectingsalivaoreggsevokeelectricalevents,re2sultingincharacteristicwaveformtypes.2.1　Characteristicwaveformscorrelatedwithparticularbe2havioraleventsoffeedingRiceplanthoppers,suchasBPHandWBPHarevascularfeedersandtargetalmostexclusivelyphloem,andrarelyxylemassuckingsites(phloemfeeders)[10,19].ThefeedingprocessofBPHconsistsoftwomajorbehavioralsteps:exploratoryprobingwiththesecretionofcoagulablesheathmaterial(salivation),andsuckingfromphloemsieveelement(phloemingestion)[20].OutofseveralwaveformsobservedduringfeedingactivitiesofBPH,threewaveforms,S,A,andIaredistinctiveandparticularlyim2portant(Fig.1).TheSwaveformwithhighamplitudethatap2pearedimmediatelyafterstyletpenetrationmayrepresentsaliva2tionofthestyletsheathaccompaniedwiththemovementsofthestyletsandsamplingsofplantfluid.TheIwaveformassociatedwiththephloemingestioncanbeconfirmedfromanalysisofhon2

Fig.1.WaveformsrecordedduringfeedingofBPHonsusceptiblerice.

Charttobereadfromlefttoright.

HATTORIMakoto:ElectronicMonitoringofFeedingandOvipositionBehaviorofRicePlanthoppersandItsApplication33

Fig.2.WaveformsrecordedduringfeedingofWBPHonrice.

　　FeedingpatternsweresummarizedwithrespecttoBPH

feedingonthreeplantswithdifferentcharacteristicsinFig.6.Onthebarnyardplant,theinsectfrequentlymadeshortprobeswithonlysalivation.Asaresult,durationofnon2probingin2creaseandsuccessfulaccessingintothephloembecamerare;namely,interruptionofprobingmainlyoccursduringsalivationinnon2phloemtissueinwhichtheantifeedantexists.Asgustatoryreceptorsarelocatedinthesmallpassagewayleadingfromthefoodducttocibarium[4],theinsectseemedtotakeupsomefluidduringsalivation.Inaphidsprobingonnon2hostplants,anab2senceofphloemingestion,butoccurrenceofnon2phloeminges2tionhasbeenfrequentlyobserved[2,15].Inthesecases,however,thepresenceorlocationofchemicalsthatcoulddeteraccessingto

Fig.3.FrequencyanddurationofprobesbyBPHonthreedifferent

plants.

Barswiththesameletterarenotsignificantlydifferent(P<0105).

thephloemoringestionwasnotdetermined.

Ontheresistantrice,theplanthoppermadethephloemac2cesswithoutdifficultyasonthesusceptiblerice,unlikeonthe

barnyardgrass.However,ingestionwasinterruptedin

Fig.4.DurationofSwaveformandpercentageofprobesconsistingofonlySwaveformduringfeedingbyBPHonthreedifferentplants.

Barswiththesameletterarenotsignificantlydifferent(P<0.05).

34ChineseJournalofRiceScience,Vol.17,Suppl,2003

Fig.5.FrequencyanddurationofIwaveformduringfeedingbyBPHonresistantandsusceptiblerice.Barswiththesameletterarenotsignificantlydifferent(P<0.05).

Fig.6.SchematicillustrationoffeedingpatternofBPHonthreedifferentplants.

thephloeminashorttime.SuchafeedinginterruptioninBPHwasalsoobservedonotherresistantricesbyusingACorDCsys2tems[11,13,23].

Thus,EMSanalysisclearlydistinguishedtwotypesofresis2tantplantswithdifferentresistancefactorsagainstplanthopperfeeding.

2.3　Characteristicwaveformscorrelatedwithparticularbe2havioraleventsofoviposition

TheEMS,combinedwithsimultaneousTVcameramoni2toringwasusedtoexplorethecorrelationbetweencharacteristicwaveformsandparticularbehavioraleventsofBPHonrice

[5]

alwaysrecordedafewminutespriortoovipositorpenetration.Thevoltagelevelincreasedrapidlywiththeovipositorpenetrationandreachedupto8timesashighasthatduringstyletpenetrationonly.DuringthishighvoltagelevelthreewaveformsSi,E,andpW,correlatingtospecificbehavioraleventswererecorded.TheSiwaveformsynchronizedwiththeforward2backwardmotionoftheovipositor,cuttingintotheplanttissue.TheEwaveformwascharacterizedbytheshapeoftheletterSwitharapidsuccessionofverylowamplitudepulses,whichcorrespondedtotheshortre2ciprocatingmotionoftheovipositortoreleaseanegg.ThepWwaveformwithasmallpeakonwhichlowamplitudepulsesweresuperimposed,occurredduringpartwithdrawaloftheovipositor.Repetitionsofthesebehavioraleventsresultedinamassofseveraleggsinarow.Afterthereleaseofthefinaleggofaneggmass,theovipositorwascompletelywithdrawn(cWwaveform)fromtheplantandthevoltagedroppedbacktotheinitiallevel.Fromasequenceofwaveformpatterns,itwaspossibletodeterminethenumberofeggsandeggmasseslaidinsidetheplant

.

Gravidfemaleswerereleasedonahalflongitudinallysectionedleafsheathtoobserveeggsinsertedintotheplanttissuefromthebackduringthemovementoftheovipositor.ObservationandrecordingofWBPHbehaviorrevealedthattheyexhibitedovipositioninamannerbasicallysimilartothatofBPH[5]withregardtosequence,andho2mologouswaveformsproducedduringoviposition(Fig.7).

Stwaveformindicatinginitiationofstyletpenetrationwas

HATTORIMakoto:ElectronicMonitoringofFeedingandOvipositionBehaviorofRicePlanthoppersandItsApplication35

Fig.7.WaveformsrecordedduringovipositionbyWBPHonrice.

Fig.8.WaveformsrecordedduringovipositionbyWBPHontheleafsheathoffully2grownrice.

tissue.

2.4　OvipositionbehaviorofWBPHonagrown2upriceplant

ItwasreportedthatWBPHdidnotpreferlayingeggsonthelowerpartofleafsheathinthefully2grownrice[7,21].Therefore,ovipositionalsequencesofgravidfemalesonrice60270daysafterseedingwereobservedbyusingEMS.Fig.8showsthatafemaletriedtolayeggsthreetimesintwodifferentsites.Although,SiandcWwaveformsrepresentingpenetrationandcompletewith2drawaloftheovipositorwererecorded,noEwaveformcorre2spondingtothereleaseoftheeggwasobserved.Thissuggestedthatovipositionbehavioritselfwasinduced,butthefullprocesswasnotcompletedduetosomedeterringfactors.Thus,theEMSwaseffectiveinidentifyingadisruptedstepinthebehavioralse2quenceonlesssuitableplantsorsitesforoviposition.Thesystemwillbeusefulinanalyzingdisruptionofthebehavioralsequenceforovipositiononresistantplants.

3　Conclusion

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