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2019—FloridaEntomologist—Volume102,No.1 59

Helicoverpaarmigera

Victor Hugo Duarte da Costa1, Marcus Alvarenga Soares1,*, Francisco Andrés

Rodriguez Dimate2, Veríssimo Gibran Mendes de Sá2, José Cola Zanuncio2,

and Fernando Hercos Valicente3,*

Helicoverpaarmigera(Hübner)(Lepidoptera:Noctuidae)larvaearepolyphagous,aggressive,andhavebeenfoundinmanyofBrazil’sagriculturalareas.Biologicalcontrolwithbaculovirusesispartofanintegratedpestmanagement(IPM)strategytomanagethisinsect.Threeisolatesofnucleo­

H.armigera (animportednucleopolyhedrovirus),andtoNucleopolyedrovirusspodoptera(SPNPV)thatwaspassedseriallythroughH.armigera

andlef­9genesrevealedthattheBrazilianisolateswerecloselyrelatedtonucleopolyhedrovirusspeciesfromAustralia,SouthAfrica,China,andIndia.TheisolatescausedhighmortalityratesinthirdinstarH.armigeralarvae.Themeanlethaldose(LD50 50

H.armigerainBrazil.

LagartasdeHelicoverpaarmigera

áreasdeproduçãoagrícola.Ocontrolebiológicodebaculovírusfazpartedomanejointegradodepragas(MIP).Trêsisoladosdenucleopoliedrovírus

(NPV)foramcoletadosempopulaçõesbrasileirasdeH.armigera ,combaseNucleopolyedrovirusspodoptera(SPNPV)emH.armigera.Foramrealizadostestes

Austrália,Índia,ÁfricadoSuleChina.TodososisoladostestadospossibilitaramocontroledelagartasdeterceiroinstardeH.armigera.Analisesbiológicasdadoseletalmédia(DL50)edotempoletalmédio(TL50)variaramentreosisoladostestados.OisoladoHearNPV­BR2apresentouos

H.armigeranopaís.

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is apolyphagous insect, and one of the most important agriculturalpestsworldwide(Cunningham&Zalucki2014).Itslarvaepreferen­tiallyfeedonthereproductivestructuresofagriculturalplants(Fitt

H.armigera areestimated tobemorethan$2billionannually,inadditiontosocio­economicandenviron­mentalcosts(Tayetal.2013).HelicoverpaarmigeranowhasspreadthroughmostofBrazil,and ispresent inParaguayandArgentina,andrecentlywasdetected(thoughnotestablished)inFlorida,USA

al.2014;Hayden&Brambila2015).

Bacillus thuringinsis (Bt)proteins,

ofH.armigera

inAustralia,China,andIndia(Tabashniketal.2009;Liuetal.2010;­

­

usedforbiologicalcontrol(Rollieetal.2013).Baculovirus­basedin­

2019—FloridaEntomologist—Volume102,No.1

­

pestssuchas Hübner(Lepidoptera:Erebidae)

andCydiapomonella(Linnaeus)(Lepidoptera:Tortricidae)(Moscardi

­

doetal.2000;Chenetal. 2002;Arrizubietaetal. 2013).This is es­

­

more,Haaseetal.(2015)reportedthatthesearchforlocalisolatesis

­

uctsbasedonforeignisolateshavebeenregisteredforcommercialuse

againstH.armigera

H.armigeralarvae.

LarvaeofH.armigerawereobtainedfromtheBiologicalControl

LaboratoryofAppliedBiologyCenter“EmbrapaMilhoeSorgo”inSete

25±2°C,with70±10%RH,anda12:12h(L:D)photoperiod,andfed

Fivevirusisolateswereobtained;3werefromH.armigeralarvae,

Threelarvae(1collectedinFeb2013and2inApr2014)showedbe­

insectsdiedwithinafewdoftheintegumentrupturing.Theother2

LC,ef­

H.armigera(HzSNPV)orBaculovirusspodop­

teraSpNPV­BR4.Onewasconsideredthereference isolate,whereas

theotherwasobtainedbyserialpassageofaBaculovirusspodoptera

SpNPV­BR4throughH.armigera

SpodopterafrugiperdaSfNPV,which

wasisolatedfromSpodopterafrugiperdaSmith&Abbot(Lepidoptera:

Noctuidae) larvae,wasobtained fromthe“EmbrapaMilhoeSorgo”

baculovirusbank,andsprayedoncornleavescontainingH.armigera

H.armigerapopula­

larvalmortality in the third passage, andwas thereforeused in the

bioassays.

­

H.armigera

9polyhedrapermLper

obtainedfromthisstepwereusedinthebioassays.

­

­

encesofthe“UniversidadedeBrasília”inBrasilia,DistritoFederal,Brazil.

­

­

­

­

totheBasicProgramLocalAlignmentSearchTool(BLAST)(Altschuletal.

­

­

­

ment. The alignments were trimmed to the size of the partial

­

4(Tamuraetal.2011).Thetreeswerebuiltby1,000replicatesof

stepwiseaddition.

HELI

COVERPAARMIGERALARVAE

H. armigera larvaewereheldwithout food

3 4 5 7 polyhedra

­

minedinadvanceusingahemocytometer(Neubauerchamber,Kasvi,

pathogen.Eachtreatmenthad32larvae.Controlleafdiscsweretreat­

.Helicoverpaarmigeramortality

hosts(Rowleyaetal.2011).

Isolated Insectspecies

HearNPV­BR1 Helicoverpaarmigera

HearNPV­BR2 Helicoverpaarmigera

HearNPV­BR3 Helicoverpaarmigera

SfNPV­BR4 Spodopterafrugiperda Cascavel,Paraná,Brazil

Helicoverpazea USA,imported

Costaetal.:BaculovirusHearNPVfromHelicoverpaarmigera

Themedian lethaldose (LD50)wascalculated foreach isolateus­

(LT50

polyhedrapermL.Thiswasthelowestdosedeterminedto

isolates.

is closely related tobaculovirusHearNPVofother regions.BR1 (Ac­

relatedtonucleopolyhedrovirusfromAustralia(HearNPV­Aus,Acces­

(AccessionNos.MF542297andMF542301)wascloselyrelatedtonu­

cleopolyhedrovirusfromSouthAfrica(HearNPV­NNg1,AccessionNo.

AP010907.2),HearNPV­Aus,HearNPV­CI(AccessionNo.AF303045.2),

notrelatedtoanyHearNPV,butwascloselyrelatedtoisolatedcontrol,

nucleopolyhedrovirus from India (HearNPV­Faridkot, Accession No.

KM357512.1)(Fig.1).

HELICOVERPAARMIGERALARVAE

H.

armigera ­

ages.TheLD50

4 4 4 5,and5

theLD50

TheBR3andBR4isolatesshowedhigherlimits,withvaluesabovethe

virulentbasedonassessmentofmortality.

TheLT50valuerangedfrom7to10d.WiththeBR1andBR2isolates,

curvereachedLT50

,BR3,andBR4iso­

­

encesbetweentheisolatesintermsofLT50values,withfasterreplica­

TherecentpresenceofH.armigerainBrazilindicatesthatitmay

2013;Costaetal.2015),commonlycollectedduringpestoutbreaks,

world(Wyckhuysetal.2013;Luoetal.2014).Naturallyoccurringiso­

latesofpathogenicH.armigerabaculovirusinBrazilcouldallowforthe

­

inSpain(Arrizubietaetal.2013),China(Luoetal.2014),andAustralia

H.armigera

­

­

manceforbiologicalcontrol(Coryetal.2005;Baillie&Bouwer2012).

­

virusisolates,asreportedforthoseusedinChinaagainstH.armigera

(Tangetal.2012).

The high degree of relatedness among BrazilianH. armigera vi­

rus isolates and those of Australia (HearNPV­Aus), China (HearNPV­

H.armigerainBrazil.

HearNPVbaculovirushadnotbeenreportedpreviouslyinthiscountry,

andthereforetheymayhavebeenintroducedbyH.armigerapopula­

pest indicates that thesemicroorganisms and their insect hosts can

overcome geographical barriers and colonizenewhabitats (Mazzi&

­

hedrovirusshowsthattheywereintroducedbyvariousmeans,asre­

portedforH.armigera

H.armigera

Thenaturaldispersionofbaculovirusispossibleduetotheirability

al.2010).Thisoccurswhenvirusesareingestedatsublethalconcen­

diseasesigns(Burdenetal.2003;Cory&Myers2003).Thelatentvirus

­

­

­

The tree is drawn to scale,withbranch lengthsmeasured in thenumberof

­

2019—FloridaEntomologist—Volume102,No.1

imposedonH.armigera

­

­

Bom­

byxmori(L.)(Lepidoptera:Bombycidae)larvaewithadultsproducing

progenyinfectedwiththesameisolateastheirancestors(Khuradet

al.2004).Similarly,30.9±2.9%H.armigeraprogenywereinfecteddue

dosesofPlodiainterpunctella(Hübner)(Lepidoptera:Pyralidae)virus

(Burdenetal.2002).

TheBR4isolatemayhaveoriginatedfromalatentbaculovirusac­

H.

armigera

Mam­

estrabrassicae(L.)(Lepidoptera:Noctuidae)whenalatentbaculovirus

Autographacalifor­

nica(Speyer)(Lepidoptera:Noctuidae)nucleopolyhedrovirusisolates

­

rusisolatesofM.brassicae)](Hughesetal.1993).Ontheotherhand,

­

ity of the alphabaculovirus isolated fromdead Lymantria dispar (L.)

tobaculoviruspreviouslyisolatedfromLymantriamonacha(L.)(Lepi­

doptera:Lymantriidae).

VirusisolatescausedthirdinstarH.armigeralarvalmortalityatthe

evaluateddoses.AnalysisofLD50valuesindicatedthattheBR2isolate

wasmorevirulentascomparedwiththeotherisolates.TheLD50values

were similar to those reported in screening tests for biological pest

controlprogramsofH.armigerainSpain(Arrizubietaetal.2013).The

H.armigeraissimilartothatreportedin

­

virusisolates(Ogemboetal.2005;Arrizubietaetal.2013).

H.armigeralar­

­

­

drovirusisolatesareinaccordancewithresultsofbaculovirusstrains

collectedintheIberianPeninsulaonsecondinstarH.armigeralarvae,

whichshowedthattheisolateHearSNPV­SP1causedmorerapidmor­

talitythantheotherIberianstrainstested(Arrizubietaetal.2013).

H.armigerainBrazilim­

­

latesinH.armigera

H.armigera.

dePessoaldeNívelSuperior(CAPES),”“ConselhoNacionaldeDesen­

­

EstudosFlorestais(IPEF)forfundingandsupport.

HelicoverpazeasinglenucleopolyhedrovirusisolatedinHelicoverpaarmig­

era

ResponsedoseandmortalityofthirdinstarHelicoverpaarmigera(Lepidoptera:Noctuidae)larvaewithnucleopolyhedrovirusisolates.

Isolate N Slope±SE X² LD50

Range P df

192 4 4 5 <0.001 4

BR1 192 4 4 5 <0.001 4

BR2 192 4 4 4 <0.001 4

BR3 192 5 5 <0.001 4

BR4 192 5 5 <0.001 4

50

thelowestandthehighestvalueforLD50;P

SurvivalcurvesofHelicoverpaarmigera

feedingoncornleavesinoculatedwith1×10 polyhedrapermLofbaculovirusLC,BR1,BR2,BR3,BR4,andincontrolobtainedwithKaplan­

Costaetal.:BaculovirusHearNPVfromHelicoverpaarmigera

­

hedrovirus isolatefromHelicoverpaarmigeraasthebasis forabiological

Helicoverpa

armigera ­

Spodopterafru­

giperda

­

Plodiainterpunc­

tella

­

­

Spodopteraexiguanuclearpoly­

­

­

Helicoverpa zea and Helicoverpa armigera

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lovirusresemblingMamestrabrassicaenuclearpolyedrosisvirusinM.bras­

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(Lepidoptera: Noctuidae) lines to a nuclear polyhedrosis virus. Environ­

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bollworm,Helicoverpaarmigera

nucleopolyhedrosisvirusthroughanalternatehostsystem.Entomophaga

alphabaculovirusisolatedfromdeadLymantriadisparlarvaeshowshigh

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nacha

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