Two new species of Drosophila (Diptera, Drosophilidae) associated with inflorescences of Goeppertia monophylla (Marantaceae) in the city of São Paulo, state of São Paulo, Brazil

RevistaBrasileiradeEntomologia62(2018)159–168

REVISTA

BRASILEIRA

DE

Entomologia

Systematics,

Morphology

and

Biogeography

Two

new

species

of

Drosophila

(Diptera,

Drosophilidae)

associated

with

inflorescences

of

Goeppertia

monophylla

(Marantaceae)

in

the

city

of

São

Paulo,

state

of

São

Paulo,

Brazil

Suzana

Casaccia

Vaz

_,

_Carlos

_Ribeiro

_Vilela

_,

_Antonio

_Bernardo

_Carvalho

a_{UniversidadeFederaldoRiodeJaneiro,InstitutodeBiologia,DepartamentodeGenética,RiodeJaneiro,RJ,Brazil} b_{UniversidadedeSãoPaulo,InstitutodeBiociências,DepartamentodeGenéticaeBiologiaEvolutiva,SãoPaulo,SP,Brazil}

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received8February2018 Accepted26March2018 Availableonline7April2018 AssociateEditor:AndrzejGrzywacz Keywords: Breedingsite Caeté Drosophilinae Feedingsite Taxonomy

a

b

s

t

r

a

c

t

TwonewBrazilianspeciesofDrosophila(subgenusDrosophila)aredescribedandillustrated:Drosophila asymmetricasp.nov.andDrosophilapeixotoisp.nov.Bothspecieswerecollected,andemerged,from inflorescencesofGoeppertiamonophylla(Marantaceae)intheurbanForestReserveoftheInstitutode BiociênciasdaUniversidadedeSãoPauloandtheirtypeswillbedepositedintheMuseudeZoologiada USP.Theformerspecies,whichcouldnotbeassignedtoanyknowngroup,hasaconspicuously asymmet-ricaedeagusandanarrowoviscaptvalve.Thelatterspeciesbelongstotheguaranigroupandisclosely relatedtoD.guaru,D.ornatifronsandD.subbadia,fromwhichitcanbedistinguishedbythepresenceof justoneconspicuouslargeblackspineatinnerlowertipofcercusinsteadoftwospines.

©2018SociedadeBrasileiradeEntomologia.PublishedbyElsevierEditoraLtda.Thisisanopen accessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

Flower-associatedspecies of Drosophila belongingto several groups within the subgenus Drosophila, suchas the bromeliae, flavopilosa,onychophoraandxanthopallescens,aswellastoother subgenera,suchasPhloridosaandSiphlodora,havebeenreported fromtheNeotropicalRegion(Brncic,1983;SantosandVilela,2005). TheDrosophilidaegeneraPalmomyia,Palmophila,Scaptomyzaand Zygothrica have also been knownto include flower-associated, Neotropical species (Malogolowkin, 1952; Brncic, 1983; Vilela, 1984; Grimaldi, 1987; Grimaldi et al., 2003; Santos and Vilela, 2005).Adaptationstoexploitsuchresourceshaveevolved inde-pendentlyseveraltimesindifferentlineages(MarkowandO’Grady, 2008).

Inarecentlypublishedarticle(Vazetal.,2014)wedescribed Drosophila calatheae, a flower-breeding species associated with inflorescences of Calatheamonophylla and C. cylindrica, vernac-ularlyknownascaeté inBrazil.ThegenusCalatheawasshown (Borchseniusetal.,2012)tobepolyphyleticandthegenus Goep-pertiawasresurrectedandredefinedtoincludeseveralspeciesof aCalatheacladeincludingC.monophyllaandC.cylindrica. There-fore, we should now refer to our previously studied breeding

∗ Correspondingauthor.

E-mail:crvilela@ib.usp.br(C.R.Vilela).

sitesasGoeppertiamonophyllaandG.cylindrica.BesidesDrosophila calatheaewealsorecognizedfouradditionalunknownDrosophila species associated to these inflorescences and less abundant regardingemergence(Vazetal.,2014;seeTables1–4).Drosophila calatheaeandDrosophilaasymmetricasp.nov.(thelattercitedby Vazetal.,2014asDrosophilasp.I4)werethetwomostcommon speciesemerging fromcollectedinflorescenceswhileDrosophila peixotoisp.nov.(citedbyVazetal.,2014asDrosophilasp.G4)was theirmostcommonvisitor.Together,thesethreespeciesaccounted for120outof 137drosophilids flies(_∼88%)thatemerged from 20inflorescencesofG.monophylla.Thefourthmostcommon vis-itor(citedbyVazetal.,2014asDrosophilasp.W3),belongingto theguaranigroup,wasrecentlydescribed(Ratcovetal.,2017)as DrosophilabutantanRatcov,VilelaandGo ˜ni.Itshouldbepointed outthatnospecimenofthelatterspeciesemergedfromthe inflo-rescencesofGoeppertiamonophylla.

Thepurposeofthisarticleistodescribetwonewspeciesbred fromG.monophylla(Figs.1–4)inflorescences:Drosophila asymmet-ricasp.nov.(ungrouped)andDrosophilapeixotoisp.nov.(guarani group).

Materialsandmethods

Methodologyregardingcollection,handlingandidentification offliesandfloralresourcesaredetailedinVazetal.(2014). https://doi.org/10.1016/j.rbe.2018.03.003

0085-5626/©2018SociedadeBrasileiradeEntomologia.PublishedbyElsevierEditoraLtda.ThisisanopenaccessarticleundertheCCBY-NC-NDlicense(http:// creativecommons.org/licenses/by-nc-nd/4.0/).

Figs.1–4.GoeppertiamonophyllaintheForestReserveoftheInstitutodeBiociênciasdaUniversidadedeSãoPaulo,SãoPaulo,SP,Brazil,on26-27.XII.2006:(1)clumpsof caetéonthebanksofthestream;(2)twoadjacentindividualsbearingoldinflorescences;(3)younginflorescencewithbuds;(4)oldinflorescencewithopenedflowersand decayingflowers.

Theaspiratedandemergedfliesaresummarizedandupdated inTable1.Dissectionsofreproductivestructureswereperformed according to Wheeler and Kambysellis (1966), as modified by Kaneshiro(1969)andBächlietal.(2004).

Table1

Drosophilidsaspiratedinnaturebetween12and28.XII.2006fromseveral inflores-cencesofGoeppertiamonophyllaandemergedinthelaboratorybetween30.XII.2006 and12.I.2007from20inflorescencesofthesamespeciescollectedintheForest ReserveofIB-USP,CidadeUniversitária“ArmandodeSallesOliveira”,SãoPaulo, stateofSãoPaulo,Brazil.a

Drosophilidspecies Aspirated

(ina5-day collection)

Emerged

Drosophilacalatheae 177 68

Drosophilapeixotoisp.nov. (citedasDrosophilasp.G4) 226 22 Drosophilaasymmetricasp.nov.(citedasDrosophilasp.I4) 24 30

Drosophilabutantan (citedasDrosophilasp.W3) 10 0

Otherdrosophilids 58 17

Total 495 137

a_{DatasummarizedandupdatedfromVazetal.(2014).}

Photomicrographs of adults were taken under 4× objective withan Olympus camera (PM2)loaded withan analog 35mm Fujichrome Professional 64T film and attached to an Olympus stereomicroscope (SZ11)bearing a ring illuminator.The analog imageswerelaterdigitizedwithanEpsonscanner(Perfection4180 photo).Photomicrographsofterminaliastructuresweretakenwith anOlympus BX60 microscope and anOlympus Q-Color 5 digi-talcamera.A10×objectivewasusedtogenerateasetof20–50 pictures bymanuallyfocusing ona given structure atdifferent depths.Thissetofphotomicrographswasthendigitallystacked tocreate an all-in-focuscomposite using theCombine ZP free software(Hadley, 2010).Outdoorphotographsweretakenwith aNikonFM-10 analogcameraloadedwithFujifilmSuperia100 ASA.

DescriptionofthenewspeciesfollowsVilelaandBächli(1990, 2000)andBächlietal.(2004,2005).Measurementsofpaired struc-turesweretakenfromtheflies’leftside.Therightwingofone doublemountedmaleofeachtypeserieswasremovedwiththe tipofanentomologicalpin,placedinadropofasolutionof70%

ethanolin5%glycerinonawellslideforatleast30min,transferred toadropofpureglycerinonamicroscopeslide,coveredwitha cov-erslip,andphotomicrographedundera4×objectiveasdetailedfor terminalia,exceptforthefactthattheywerenotstained.Labeldata attachedtoeachtypespecimenarecitedinfullwithcomma desig-natingalinechange,slashesindicatingalabelchangeandclarifying notesincludedinbrackets.Typeseriesofbothnewspecieswillbe depositedintheMuseudeZoologiadaUniversidadedeSãoPaulo (MZSP).

D.peixotoisp.nov.wasrearedforseveralgenerationsinthe laboratoryusingaflowerenrichedmediumdescribedinVazetal. (2014).

Results Taxonomy

GenusDrosophilaFállen,1823:4.

SubgenusDrosophilaSturtevant,1942:30.

Drosophila(Drosophila)asymmetricasp.nov.(Figs.5–33)

Drosophilasp.I4,Vazetal.,2014:608(Table1,feedingsite),609 (Fig.6,puparium[upperone]),611(Tables3and4,breedingsite), suppl.keyS1(feedingsite,key),suppl.keyS2(breedingsite,key). Diagnosis.Medium-sizeddullgrayishyellowfly,bodylength (excludingwing) about2.7mm(male) and 3.0mm (female); h index1.1–1.8;midkatepisternalabout13–40%oftheanteriorone; abdomendull, grayishyellow;tergites 2–5withmedially inter-ruptedposteriordarkerbands;tergites1and6devoidofbands; wings uniformlyslightly darkened;C index 2.0–3.0;larvae and pupaewithverylongconspicuousposteriorand anteriorhorns, respectively;aedeagusdistallyasymmetric;conspicuousnarrow oviscaptvalve.

Materialexamined(10males,7females;depositedinMZSP). Holotypemale [wild-caught,coded M18,double-mounted, pho-tomicrographed(Figs.5–7),rightwingremoved(Fig.11),dissected (Figs.12–26)]labeled:“Brasil–SP–SãoPaulo,ReservaFlorestal

S.C.Vazetal./RevistaBrasileiradeEntomologia62(2018)159–168 161

Figs.5–10.Drosophilaasymmetricasp.nov.,typespecimens:(5–7)maleparatype,leftlateral,obliquedorsalanddorsalviews,respectively;(8–10)femaleparatype,left lateral,obliquedorsalanddorsalviews,respectively.Scalebar=1mm.

Fig.11.Drosophilaasymmetricasp.nov.:maleholotype,rightwing,dorsalview.Scalebar=1mm.

do Instituto de Biociências, 28.XII.2006, Vilela coll./Drosophila

asymmetrica [male symbol], Vaz, Vilela &

Car-valho/fotomicrografado[photomicrographed]/termináliailustrada [illustratedterminalia]/HOLOTYPE/[microvialwithterminaliaand rightwinginglycerin]”.Sevenparatypes(5♂♂,2♀♀),samedata as holotype, plus 9 paratypes (4 ♂♂ [one photomicrographed (Figs. 5–7)],5♀♀[onephotomicrographed(Figs.8–10),one dis-sected(Figs. 27–33)]),same data as holotypeexcept collection date(18,22,26.XII.2006).

Type locality. Forest Reserve of the Instituto deBiociências daUniversidadedeSãoPaulo(IB-USP)(23◦33.96S;46◦43.72W), CidadeUniversitária“ArmandodeSallesOliveira”,SãoPaulocity, stateofSãoPaulo,Brazil.

Description.Male (n=10). Headand frons dull grayish yel-low,frontallength0.37mm(0.34–0.39)mm;frontalindex=1.12 (1.00–1.23),toptobottomwidthratio=1.56(1.46–1.69).Frontal trianglelight graypollinose,about100%offrontallength; ocel-lar triangle shiny light brown, 31–43% of frontal length. Or3 to or1=67–83% of that to inner vertical; or1/or3 ratio=0.95 (0.78–1.22); or2/or1 ratio=0.50 (0.36–0.71); poc 50–60%, oc 56–73%offrontallength;vibrissalindex=0.14–0.40,vtindex=1.12 (1.00–1.33),facial carinalightbrown, nose-like,slightly sulcate. Genalightbrown,cheekindexabout5.50–8.00.Eyedarkred.Eye index=1.10–1.40.Pedicelbrown,laterallylighter,firstflagellomere lightbrown, lengthtowidthratio1.20–2.00.Aristawith3 dor-sal,1–2ventraland3–4tinyinnerbranches,plusterminalfork. Proboscisbrown.

Thoraxdullgrayishyellow;length1.11(1.00–1.22)mm.Scutum pollinose,brownish-gray,8rowsofacrostichalsetae.hindex=1.32 (1.11–1.80).Transversedistanceofdorsocentralsetae251%of lon-gitudinaldistance;dcindex=0.66(0.58–0.72).Scutellumpollinose, laterally light brown, medially darker; distancebetween apical scutellarsetaeabout71%ofthatofapicaltobasalone,basalsetae convergent;apicalsetaecruciate,scutindex=0.81(0.68–0.89). Hal-tergray.Pleurabrownishgray,sternoindex=0.78(0.59–0.88),mid katepisternalsetaabout13–40%oftheanteriorone.Legsbrownish gray.

Wing (Fig. 11) uniformly slightly darkened; length 2.06 (1.78–2.67)mm,lengthtowidthratio=2.06(1.93–2.21).Indices: C=2.60 (1.95–2.84), ac=2.46 (2.11–2.71), hb=0.49 (0.40–0.63), 4C=0.90(0.79–1.16),4v=1.69(1.50–1.91),5x=1.75(1.67–1.86), M=0.54(0.46–0.58),prox.x=0.59(0.52–0.68).

Abdomen (Figs. 5–7) dull grayish yellow; tergites 2–5 with medially interrupted posterior darker bands; tergites 1 and 6 devoid of bands. Male Terminalia (Figs. 12–26). Epandrium microtrichoseonposteriormediananddorsalareas;ca.2upper and12lowersetae;ventrallobeweaklysclerotized,slightly cover-ingsurstylus,devoidofmicrotrichiae,posteriormarginnoticeably concave. Cerci mediodorsally microtrichose, widely fused to epandrium. Surstylus bearing a small patch of microtrichiae on mediodorsal area, with ca. 7 cone-shaped prensisetae, ca. 4 long, strong outer setae and ca. 11 long, thin, mostly inner setae.DecasternumasinFigs.13,14,23;anteriorlyturned dor-salwards, anteromedial tip area slightly sclerotized (not seen

Figs.12–26. Drosophilaasymmetricasp.nov.,maleholotype,terminalia:(12–14)epandrium,cerci,surstyli,anddecasternum,leftlateral,obliqueposteriorandposterior views,respectively;(15–17)hypandrium,gonopodsandparaphyses,leftlateral,obliqueposteriorandposteriorviews,respectively;(18–22)aedeagus+aedeagalapodeme, severalviewsfromdorsalthroughventral;(23)epandrium,cerci,surstyli,anddecasternum;ventralposteriorview;(24–26)ejaculatoryapodeme,dorsal,obliquedorsaland leftlateralviews,respectively.

in Figs. 13, 23), sharplycarinate on posterior mediodorsalhalf (Fig.14).Hypandrium(Figs.15–17)shorterthanepandrium, ante-riormarginconvex;dorsalarchandposteriorhypandrialprocess absent;gonopodbare,linkedtoparaphysisbymembranoustissue (Fig.17).Aedeagus(Figs.18–22)mediallyanddistallyasymmetric, anteriorlybearingapairoftiny,acutespinespointedbasalwards, subdistallybearing pair ofdorsal, small, triangle-shaped spines (Fig.18);rightsidemediallyanddistallypleatedleftwardsover mostdorsalcleft,partiallycoveringleftside,hidingleftsubdistal spine.Aedeagus in dorsal (Fig. 18)and ventral(Fig. 22) views depictingonlytherightsubdistalspineontheleftside;apically bluntin lateralview (Fig.20).Aedeagal apodeme,rod-shaped, curved,laterallyflattened,slightlyshorterthanaedeagus.Ventral rod completely fused to aedeagal apodeme, distally expanded laterally,relativelylong(Fig.20).Paraphysisright-angled triangle-shapedin lateral view(Fig.16), bearing a small setula at very distal tip. Ejaculatory apodeme as long as aedeagal apodeme,

dorsodistallyexpandedlaterallyandcrescent-shaped(Figs.24,25), laterally L-shaped and proximally expanded dorsoventrally (Fig.26).

Female(n=7).Headandfronsdullgray,frontallength0.40mm (0.37–0.41) mm; frontal index=1.12 (1.00–1.21), top to bot-tom width ratio=1.65 (1.47–1.79). Frontal triangle light gray pollinose, about 100% of frontal length; ocellar triangle shiny light brown, 35–40% of frontallength. Or3 toor1=67–100% of that to inner vertical; or1/or3 ratio=0.88 (0.82–0.91); or2/or1 ratio=0.53(0.44–0.60);poc53–67%,oc71–81%offrontallength; vibrissalindex=0.21–0.42;vtindex=1.07(1.00–1.18);facialcarina lightbrown, nose-like,slightly sulcate.Genalightbrown, cheek index about 5.75–8.33. Eye dark red. Eye index=1.14–1.42. Pedicel brown, laterally lighter, first flagellomere light brown, length to width ratio 1.40–2.00. Arista with 3 dorsal, 2 ven-tral and 2–5tiny inner branches, plus terminal fork. Proboscis brown.

S.C.Vazetal./RevistaBrasileiradeEntomologia62(2018)159–168 163

Figs.27–32.Drosophilaasymmetricasp.nov.,femaleparatype(notthespecimeninFigs.8–10),terminaliaandspermathecae:(27–30)tergite8,epiproctandhypoproct,left lateral,obliqueposterior,posteriorandventralposteriorviews,respectively;(31)leftoviscaptvalve,outerlateralview;(32)spermathecaeinnercapsules,lateralview.Scale bar=0.1mm.

Fig.33.Drosophilaasymmetricasp.nov.,femaleparatype,terminalia:leftoviscaptvalve,outerlateralview.Scalebar=0.1mm.

Thoraxdullgrayishyellow;length1.27(1.20–1.39)mm.Scutum pollinose,brownish-gray,8rowsofacrostichalsetae.hindex=1.27 (1.11–1.50).Transversedistanceofdorsocentralsetae242%of lon-gitudinaldistance;dcindex=0.66(0.58–0.74).Scutellumpollinose, laterally light brown, medially darker;distance betweenapical scutellarsetaeabout78%ofthatofapicaltobasalone,basalsetae convergent; apical setae cruciate, scutindex=0.85 (0.81–0.90). Haltergrayishyellow. Pleuragrayishyellow,sterno index=0.80 (0.74–0.83),midkatepisternalsetaabout21–33%oftheanterior one.Legsdullgrayishyellow.

Wing uniformly slightly darkened; length 2.34 (2.20–2.54) mm, length to width ratio=2.08 (2.02–2.14). Indices: C=2.70 (2.42–2.95),ac=2.42(2.20–2.67),hb=0.53(0.40–0.64),4C=0.89 (0.79–1.00),4v=1.66(1.52–1.96), 5x=1.51(1.33–1.75),M=0.52 (0.44–0.61),prox.x=0.58(0.52–0.63).

Femaleterminalia (Figs.27–33).Tergite 8narrow, devoidof setae(Fig.27),anterodorsallymicrotrichose,mediodistallyweakly sclerotized(Fig.28);ventralhalfcurvedfrontwards,distally bear-ingarowofca.8thin,longsetulaeadjacenttoposteriormargin (Figs.27–30);epiproctmostlymicrotrichose,setose, anterolater-allyembracedbybasalareaoflargerhypoproct(Fig.29);hypoproct mostlymicrotrichose,setose,conspicuouslybearingtwopairsof strongerandlongersetae(Fig.27).Valveofoviscaptrelatively nar-row,distallyrounded,subdistallyslightlyexpandeddorsoventrally andventrallyslightlysinuateinlateralview(Figs.31,33),bearing ca. 5 discal and about20 marginal, peg-like,mostly roundish-tipped,outerovisensilla(Fig.33);trichoid-likeinnerovisensilla: 3thindistallypositioned,and1long,slightlycurved,subterminal (Fig.33).Spermathecalinnercapsulecylinder-shaped,sclerotized, enlargedatbase,distallyflattened;basalintrovertdeeply invagi-nated(Fig.32).

Puparium.OnepupariumofDrosophilaasymmetrica,sp.nov. col-lectedfromaninflorescenceofG.cylindrica,inRiodeJaneirocity wasphotomicrographed(Fig.6[upperone]ofVazetal.,2014[cited thefigurecaptionasDrosophila sp.I4];hostplant unintention-allyomitted)togetherwithonepupariumofDrosophilacalatheae [lowerone],obtainedfromthefromthesamehostplant.Puparia ofbothspeciesconspicuouslybearlonghornsandawideair-filled spaceontheiranteriorend,probablyrelatedtofloatationinside bractsphytotelmata.

Ecology. Breeds on inflorescences of Goeppertia monophylla (Table1).

Etymology.Namedtoindicateitsconspicuously asymmetric aedeagus.

Note. A remarkablysimilar oviscaptvalve and similar sper-mathecaeweredepictedandphotomicrographedbyVilela&Bächli (1990:215,Fig.58EFig.58;323,Fig.166B)forthenoticeablysmaller femaleparalectotypeofDrosophilabolivianaDuda,1927,collected from“YungasvonCoroico”,Bolivia.Thecitedauthorsconsidered thetwofemaleparalectotypesofthetypeseriesofD.bolivianaas belongingtodistinctspecies.Aswesuspectthesmallerfemale par-alectotypeofDrosophilabolivianaandDrosophilaasymmetricacould beconspecific,theeventualcaptureandidentificationofasimilar malefrom“YungasvonCoroico”wouldbedesirabletoconfirmour suspicion.

Drosophila(Drosophila)peixotoisp.nov.(Figs.34–56).

Drosophilasp.G4,Vazetal.,2014:608(Table1,feedingsite), 610(Table2,feedingsite),611(Table3,breedingsite),suppl.key S1(feedingsite,key),suppl.keyS2(breedingsite,key).

Figs.34–39. Drosophilapeixotoisp.nov.,typespecimens:(34–36)maleholotype,leftlateral,obliquedorsalanddorsalviews,respectively;(37–39)femaleparatype,left lateral,obliquedorsalanddorsalviews,respectively.Scalebar=1mm.

Fig.40. Drosophilapeixotoisp.nov.:maleparatype,rightwing,dorsalview.Scalebar=1mm.

Diagnosis.Small-sized dullbrownfly, witha relativelylarge headinlateralview,bodylength(excludingwing)about2.1mm (male) and 2.2mm (female), front brown, ocellar triangle dark brown;scutumpollinose,lightbrownanteriorlyandbetween dor-socentrals,graduallydarkeningtowardlateralanddistalregions; scutellumdulldark brown;setaeand setulaedarkbrownwith a remarkablegolden sheen; wings light brown hyaline, tips of veinsR2+3,R4+5andMslightlyclouded,crossveinsslightlyclouded

(dM-Cudarker),lappetbearingonethickandonethinsetaattip; abdominaltergitesshiny;maleterminaliaconspicuouslybearinga largeblackspineatinnerlowertipofcercus;oviscaptvalvedistally shortandblunt,submarine-shaped.

Material examined (10 males, 10 females, deposited in

MZSP).Holotypemale(wild-caught,codedM18,double-mounted, right wing removed, dissected [Figs. 41–53]) labeled: “Brasil – SP – São Paulo, Reserva Florestal do [Forest Reserve of] IB-USP, Cidade Universitária, 26.XII.2006, Vilela coll./Drosophila peixotoi[male symbol] Vaz, Vilela & Carvalho/fotomicrografado [photomicrographed]/terminália ilustrada [illustrated termina-lia]/HOLOTYPE/[glass microvial with terminalia and right wing in glycerin]”. Paratypes (9 ♂♂ [one with right wing removed (Fig.40)],10♀♀[onephotomicrographed(Figs.37-39)anddissected (Figs.54–56)]),samedataasholotype.

Type locality. Forest Reserve of theInstituto de Biociências daUniversidadedeSãoPaulo(IB-USP)(23◦33.96S;46◦43.72W), CidadeUniversitária“ArmandodeSallesOliveira”,SãoPaulocity, stateofSãoPaulo,Brazil.

Description.Male(n=10).Headrelativelylargeinlateralview, brown. Frontallength0.26 (0.24–0.29)mm, frontalindex=0.98 (0.83–1.00),toptobottomwidthratio=1.33(1.17–1.50).Frontal triangle laterally shiny, 60–80%of frontal length;ocellar trian-gle shiny dark brown, 40–50% of frontal length. Orbital plates shiny, 75–91% of frontal length. Orbital setae black, or2 out-side or1 and or3; distance of or3 to or1=75% of or3 to vtm, or1/or3 ratio=0.85, or2/or1 ratio=0.38, postocellar setae=52 (50–73)%,ocellarsetae=78(67–91)%of frontallength; vtindex 0.92(0.78–1.00);vibrissalindex0.88(0.83–1.00).Faceandcheek lightbrown.Carinalightbrown,prominent,narrow,notsulcate. Cheekindex6–10.Eyeindex=1.28(1.18–1.58).Scapeandpedicel lightbrown,firstflagellomerebrown,lengthtowidthratio=1.68 (1.50–2.00).Aristawith4–5dorsal,2ventrallongbranches,and 4–6innerbranchesrelativelylong,plusterminalfork.Proboscis andpalpusbrown.

Thoraxsubshining,mostlybrown;length0.87mm;6rowsof acrostichals.Transversedistanceofdorsocentralsetae213%of lon-gitudinaldistance;dcindex0.64.Scutellumpollinosedarkbrown, apicallyblunt;distancebetweenapicalscutellarsetae100–150% ofthatbetweenapicalandbasalone;scutindex=0.94(0.80–1.07); basalsetaedivergent,apicalonescruciate.Pleurasubshiningdark brown,sternoindex=0.48,mediankatepisternalseta33–67%of anteriorone.Halterlightbrown.Legsuniformlylightbrown;apical setaonprotibiaandmesotibia;preapicalsetaonalltibiae.

Wing(Fig.40)light brown, crossveinsR-M and dM-Cu, and apices of veins R2+3, R4+5 and M slightly clouded; length 1.82

S.C.Vazetal./RevistaBrasileiradeEntomologia62(2018)159–168 165

Figs.41–53. Drosophilapeixotoisp.nov.,maleholotype,terminalia:(41–43,52)epandrium,cerci,surstyli,anddecasternum,leftlateral,obliqueposterior,posteriorand ventralposteriorviews,respectively;(44–46,53)hypandriumandgonopods+paraphyses,leftlateral,obliqueposterior,posteriorandanteriorviews,respectively;(47–51) aedeagus-aedeagalapodeme,severalviewsfromdorsalthroughventral.Scalebar=0.1mm.

Figs.54,55.Drosophilapeixotoisp.nov.,femaleparatype,spermathecaeandterminalia:(54)spermathecaeinnercapsules,lateralview;(55)leftoviscaptvalve,outerlateral view.Scalebar=0.1mm.

(1.71–1.95)mm,lengthtowidthratio=2.10(2.03–2.19).Indices: C=2.94 (2.72–3.13), ac=2.13 (1.88–2.43), hb=0.30 (0.25–0.38), 4C=0.82 (0.76–0.89),4v=1.67(1.59–1.83),5x=1.48(1.29–1.67), M=0.49(0.43–0.55),prox.X=0.53(0.45–0.67).

Abdomen (Figs. 34–36) shiny brownish, tergite 1 yellow, tergites2–4anteromediallyyellowwithadistaldarkbrownband,

mediallyinterrupted and laterally broadened,reachinganterior marginoftergite;tergite4bearinganirregularcoffeebrownspot onthemedianregionoftheyellowarea;tergites5–6entirelycoffee brown(tergite5anteriorlylightersubmediallyinsomespecimens). Male Terminalia (Figs. 41–53). Epandrium (Figs. 41–43, 52) almostbare,slightlymicrotrichoseonposteriordorsalarea;upper

setae absent; 4–5 lower setae in a vertical row; ventral lobe notcoveringsurstylus.Cercusmicrotrichoseondorsocentralarea, conspicuously bearing a strong,long (ca.2/3 lengthof cercus), slightlycurvedspineatinnerlowertip,linkedtoepandriumby membranoustissue (Fig.43);thecurved spinebearsa longitu-dinalline along most of its lengththat could bean indication that it has originated fromthe fusion of two spines. Surstylus notmicrotrichose,withabout8–9cone-shapedprensisetae,about 10–12long,strongoutersetaeandabout9long,thin,mostlyinner setae.Decasternumupper-positioned,asinFig.52.Hypandrium (Figs. 44–46, 53) as long as epandrium, anterior margin con-vex;posteriorhypandrialprocessabsent;dorsalarchW-shaped, strongly sclerotized (Figs. 46, 53); gonopod not microtrichose, linkedtoparaphysisbymembranoustissue,bearingonelongseta onanteriorinnermargin.Aedeagus(Figs.47–51)distallybifid(in dorsal and ventralviews, Figs. 47, 51), distal 1/3laterally cov-eredwith many spines, marginally serrated ventrally (Fig. 50), turnedabruptlydorsad(Fig.49)andbearingadorsoventral lap-pet, mediodorsally with a pair of membranous, anterodorsally slightlysclerotized,finger-shaped,backwardsdirected, conspicu-ousprocesses(Figs.48–50),whicharestronglyexpandedlaterally astwotriangle-shapedstructures(indorsalview,Fig.47),shorter thanaedeagus(ca.1/3itslength)andlateroventrallycoveredwith tinyspines;dorsalcleft restrictedtoatinyopeningadjacentto thefusionaedeagus-aedeagalapodeme;paraphysisdorsodistally swollen,not microtrichose, bearing a setula onposterior inner margin just adjacentto swollen area(Figs. 45, 46, 53). Aedea-galapodemerod-shaped,aslongasaedeagus;anteriorlyslightly expandeddorsoventrally(Figs.49,50).Ventralrodtriangle-shaped, completelyfusedtoaedeagalapodeme(Fig.49).

Female(n=10).Colordifferencefrommale:insomespecimens, colorpatternoftergite5issimilartothatofmaletergite4.

Measurements: Frontal length0.26mm; frontal index=0.87, toptobottomwidthratio=1.25.Frontaltriangle60–80%offrontal length.Ocellar triangle 33–50% of frontallength. OrbitalPlates 80–100%of frontallength. Distance of or3 to or1=60–100% of or3tovtm,or1/or3ratio=0.88 (0.75–1.12),or2/or1 ratio=0.31 (0.22–043), postocellar setae=69 (64–75)%, ocellar setae=75 (64–80)% of frontal length; vt index=0.99 (0.88–1.12); vib-rissal index=0.78 (0.71–0.86). Cheek index 9.75 (9.00–10.50). Eye index=1.30 (1.20–1.38). First flagellomere length/width ratio=1.62(1.25–1.75).

Thoraxlength0.93(0.88–1.00)mm.hindex=0.90(0.62–1.00). Transverse distance of dorsocentral setae 171–260% of longi-tudinal distance; dc index=0.68 (0.56–0.79). Distance between apicalscutellarsetae100–125%ofthatbetweenapicalandbasal one;scutindex=0.92(0.87–1.00),sternoindex=0.57(0.46–0.69), mediankatepisternalseta46–69%ofanteriorone.Winglength1.98

(1.82–2.10)mm,lengthtowidthratio=2.16(2.08–2.22).Indices: C=3.19 (2.89–3.44), ac=2.00 (1.78–2.43), hb=0.29 (0.24–0.32), 4C=0.77 (0.71–0.82),4v=1.67(1.50–1.84),5x=1.49(1.22–1.83), M=0.49(0.42–0.55),prox.x=0.52(0.46–0.59).

Femaleterminalia(Figs.54–56).Valveofoviscaptdistally rel-ativelyshort, submarine-shaped in lateralview,double-walled, innerwall(Fig.56,dottedline)ca.2/3narrowerthanouterwall, api-callyroundish,submediallyslightlyexpandeddorsadlookinglike asubmarinesail,ventrallystronglyconvex,withca.14discaland about11marginal,peg-like,mostlyroundish-tipped,outer ovisen-silla;trichoid-likeinnerovisensilla:3thindistallypositioned,and 1long,slightlycurved,subterminal.Spermathecalinnercapsule lightbulb-shaped,sclerotized,notfurrowedatbase;basalintrovert deeplyinvaginated.

Ecology. Breeds on inflorescences of Goeppertia monophylla (Table1).

Etymology. Named after Dr. Alexandre Afrânio Peixoto

(1963–2013)forhisresearchcontributionsonDrosophila genet-ics,evolution,andbehavior(LaboratóriodeBiologiaMolecularde Insetos,Fiocruz,RiodeJaneiro,Brazil).

Note.FiveisofemalelinesofDrosophilapeixotoisp.nov.were establishedbyFlaviaJ.Krsticevicfromfliesaspiratedfrom inflo-rescencesofGoeppertiacylindricaofaprivatebackyardgardenin SantaTeresadistrict,RiodeJaneiro.Thefiveisofemalelines(coded STA-KF7,STA-KF8,STA-KF9,STA-KF11,STA-KF12)wererearedfor severalgenerationsonaG.cylindricaenrichedmediumdescribed inVazetal.(2014),andsamplesarepreservedinethanolinthe lab-oratoryofoneofus(ABC).However,itshouldbepointedoutthat noadultsofDrosophilapeixotoisp.nov.emergedfromG.cylindrica inflorescencescollectedintheJardimBotânicodistrict(22◦57.95S, 43◦14.31W),adjacenttoParqueNacionaldaTijuca(RiodeJaneiro city),althoughtheywerefrequentlyfoundfeedingonthese inflo-rescencesinnature(Vazetal.,2014:610,611[Tables3,4]). Discussion

Asymmetry in male terminalia. Left-right asymmetry in

the male terminalia appeared several times independently in the evolutionary history of insects and has been associated withchangesinmatingpositions(Huber, 2010).Indrosophilids asymmetry is not common but also appeared independently severaltimes,suchasinthecanalinea,flavopilosaandnannoptera speciesgroups (Vilela,1984; VilelaandBächli,1990;Vilela and Pereira, 1992; Vilela et al., 2008; Lang and Orgogozo, 2012). Drosophila pachea, belonging to the nannoptera group, has a conspicuous asymmetry in its epandrium lobes, with the left lobe being about 50% longer and thinner than the right lobe. Copulation requires a characteristic one-sided mating posture

S.C.Vazetal./RevistaBrasileiradeEntomologia62(2018)159–168 167

(Lang and Orgogozo, 2012). The asymmetry we describe in Drosophilaasymmetricasp.nov.islimitedtotheaedeagus,similar totwospeciesoftheflavopilosagroup:Drosophilamariaehelenae Vilela,1984andDrosophila hollisaeVilelaandPereira,1992.We have not made any observationoncopulation postures in this species.

TaxonomichistoryoftheDrosophilaguaranispeciesgroup. TheguaranispeciesgroupwasfirstrecognizedbyDobzhanskyand Pavan(1943)andincludedfourdescribedspecies(D.guarani,D. guaru,D.guaramunuandD.griseolineata).King(1947)addedD. sub-badiaandD.guarajatothegroupandproposeditsdivisionintotwo subgroups:guaramunuandguarani.Henotedthatthemalecerci oftheguaranisubgroup(thenincludingD.guarani,D.guaruand D.subbadia)bear“heavilypigmentedandpeculiarlycurved bris-tleswhichlooklikebentspikes”.Astoguaramunusubgroup(then includingD.guaramunu,D.griseolineataandD.guaraja)noclear morphologicaldiagnosiswasproposed.Kastritsis(1969)later sug-gestedthesetwosubgroupsshouldberaisedtothespeciesgroup statusduetocharacteristicsonpolytenechromosome morphol-ogythatapproximatespeciesoftheguaramunusubgrouptosome speciesof thetripunctataspecies group.In fact,theguaramunu subgroupseemstobeanevolutionarytransitionbetween tripunc-tataandguaraniregardingnotonlychromosomalmorphologybut internalandexternalanatomy(Throckmorton,1962;Frota-Pessoa, 1954)anddataonmolecularphylogeny(Hatadanietal.,2009;Van derLindeetal.,2010).Asmorespeciesrelatedtothegroupwere describedthelimitsbetweenguaraniandguaramunusubdivision becameunclearandspeciesweremainlyassignedtoguaraniasa speciesgroup.AtaxonomicreviewbyValetal.(1981)discussesthe difficultyofplacingspeciesineitherguaraniorguaramunu divi-sionsandtreatsguaraniasaspeciesgroupcomprisingatotalof 10species.Theyalsodiscusssomeofthemaleterminalia hetero-geneityoftheguaramunusubgroupanditsrelationtotripunctata. Ina morerecent reviewofNeotropicaldrosophilids (Vilela and Bächli,1990)D.guaramunuwasconsideredajuniorsynonymof D.maculifronsandD.guaraniajuniorsynonymofD.ornatifrons, butthegroupnamewasmaintainedasguarani.Inthatsamework, D.neoguaramunuwastransferredtothetripunctatagroup.These authorsreinforcedthesimilaritybetweenspeciesfromtheguarani subgroupandevenraisedthepossibilityofsynonymybetweenD. subbadia(describedfromthestateofMichoacán,Mexico)andD. guaru(describedfromthestateofSãoPaulo,Brazil).Theystated “furtheranalysisusingstrainsfromalltherangeofdistributionof bothspecieswillprobablyrevealthattheyindeedbelongtothe samebiological speciesor speciesinstatunascendi” (Vilela and Bächli,1990:79).

Newtaxonomic proposal. The guaranispecies group sensu

latoisclearlynot monophyleticmainlyduetoheterogeneityof theguaramunusubgroupanditsoverlappingwiththetripunctata speciesgroup.However,asdiscussedpreviously,theguarani sub-groupseemshighlyhomogeneous.Thereforeweproposethat:(I) theguaramunuandguaranispeciesgroupssensuKastritsis(1969) nottobeadopted,andtheguaranispeciesgroupbemaintained untilmoredetailsofitsrelationtotripunctataandothergroups isavailable; (II)the guaramunusubgroupbedismissed andthe guaranisubgroupbemaintainedbutrecircumscribedinanarrower senseaccordingtoKing(1947).Theguaranisubgroupwouldthen includethefollowingspecies:D.ornatifrons,D.guaru,D.subbadia andD.peixotoisp.nov,sharingthefollowingpresumed synapo-morphy:presenceofoneortwopairsofconspicuouslargeblack spinesonventralcerci.Accordingly,D.tucumanaandD.urubamba areexcludedfromtheguaranisubgroup.Accordingtothisnew proposaltheguaranispeciesgroupcurrentlywouldcomprise19 species(Ratcovetal.,2017;Bächli,2018;thiswork)ofwhich4 belongstotheguaranisubgroupandtheremaining15have uncer-tainrelationships.

BreedingsitesintheDrosophilaguaranispeciesgroup.Most speciesofthegroupweredescribedbasedonadultfliesattracted tobanana-baitedtrapsanddataregarding naturallarval breed-ing sites are scarce. D. griseolineata, D. maculifrons(cited as D. guaramunu),and D.ornatifrons(citedasD.guarani)seemstobe the most polyphagous species emerging from several types of fruitsandinfructescences(ValenteandAraújo,1991).D.guaraja was reared frominfructescences of Philodendron bipinnatifidum (Araceae;Vilela,2001).Pipkinetal.(1966)citedoneunidentified speciesofthegroup(guaranisp.)emergingfromflowersof Eryth-rinaberteroana(Fabaceae).AlongwithD.peixotoisp.nov.wealso identifiedD.griseolineataemergingfrominflorescencesof Goep-pertiamonophylla,althoughlessabundantly(7vs.22D.peixotoisp. nov.specimensoutof137drosophilids;Vazetal.,2014).Sofar,D. peixotoisp.nov.seemstobetheonlyexclusiveandhost-specific flowerbreedingspeciesofitsgroup.Studiesregardingemergence offliesfromflowersandinflorescencesofdifferentplantspecies are neededtodetermine thebreedingsites ofthose species of Drosophilathatarerarelyaspiratedornetsweptoverfruit-baited traps.

Conflictsofinterest

Theauthorsdeclarenoconflictsofinterest. Acknowledgements

WeareindebtedtoDeniseS.Sheepmakerforallowingtheuse ofherstereophotomicroscopeandtoFlaviaKrsticevicforhelpwith labandfieldwork.Wethankananonymousreviewerforhiscareful readingofourmanuscriptandsuggestionsthatimprovedthe sub-mittedversion.ThisworkwassupportedbyConselhoNacionalde DesenvolvimentoCientíficoeTecnológico–CNPq,Coordenac¸ãode Aperfeic¸oamentodePessoaldeNívelSuperior–CAPES,Fundac¸ão deAmparoàPesquisadoEstadodoRiodeJaneiro–FAPERJand WellcomeTrust(grant207486/Z/17/Z).

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