Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. iers onwbhvoso hrceitc nteaatdseista ol raea nrcdne pest. unprecedented an create could may that changes species adaptive adapted such Eventually, the in 2012). characteristics playing al., thus or et species, behaviors recipient (Pardo-Diaz new a the to radiations is Tonzo, in rise adaptive hybridization changes 2013; give in adaptive introgressive al., prompt role Moreover, et may important (Martin that 2020). an variation species al., genetic sympatric interspe- of et young following as Valencia-Montoya source introgression in to potential 2020; as role referred Ortego, such important and processes, & an reported Other play Papadopoulou, also been species. associations may have co-occurring plant hybridization, 2004) of host of cific Silvain, and fate roles & 2004), the the McMichael, al., shaping context, Prowell, good et this processes (Frati 2020; considered In infections al., be endosymbiotic 2012). et 2007), (Nosil, can (Poveda-Martinez al., selection ranges divergent et distribution (Santos of timing overlapping means by mating with boundaries. speciation insects ecological species herbivorous of maintain diversification candidates of that drive mechanisms species that the related processes and Closely the levels allopatric on evolu- interspecific and information in and sympatric provide interest intra- in can the special patterns species at of structure divergent are genetic recently population species of of related populations study closely the in Furthermore, diversity biology. genetic tionary interspecific intra-and of Patterns across flow in gene Introduction occurred limiting interchange may divergence genetic baumannii during of C. role events on important historical feeding an indicated played population results likely moth our plants the the Overall, host species. that for but showed bucyrus. host moths, and intraspecific C. a cactophagous revealed cactorum to also as Cactoblastis C. survey conspecific hybridization. reported Our and as in of never flow. doddi considered gene as plant footprints C. historical be overlap, a of for both mitochondrial signs baumannii, ranges in searched detected and and structure Cleistocactus their and SNPs species geographic on which delimited moths genome-wide well feeding in three different cactophilic of identified population areas have We these combination moth genus. are species of a a there These patterns evaluated Using However, we phylogeographic plants. 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Cactoblastis are interspecific and species and diverged doddi recently intra of driving Cactoblastis populations processes allopatric and the sympatric understand natural in to variation genetic of patterns of Surveys Abstract 2021 15, March 4 3 2 1 Logarzo Guillermo Poveda-Mart´ınezDaniel sympatric partially species in moth variation cactophagous genomic related host and Spatial OIE-NVRIA EBEO AIRES BUENOS DE CONICET-UNIVERSIDAD Agriculture-ARS of Department Invasivas U.S. Especies de Estudio el Fundaci´on Aires para Buenos Ecolog´ıa, Evoluci´on Gen´etica de de y Instituto 2 n sea Hasson Esteban and , 1 ar Varone Laura , 2 aeaFetsCorona Fuentes Malena , 4 1 2 tpe Hight Stephen , 3 , Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. .ficus-indica O. hsnwfidnscudepn h itbedho h led ecie pce n/rtenme of number the and/or species 2014). al., described et already Varone the 1985; of McFadyen, breadth 1969; diet Mann, the 1999; expand Neder, could & Cactoblastis findings (Arce larvae new and This adults on markings genus species on cactorum feeding reported been baumannii has Cleistocactus also from but different 2021) appeared Varone, genera & of the Soto, use Esteven, of the Otegui, (Barbarich, confirmed NWA is surveys in interest Recent NWA. of McFadyen, to species & Argentina third Erb, The Zimmermann, 2007). 1985). 1985; (McFadyen, Walton, (McFadyen, NWA & to (Raghu western 1979). industry central dye from provinces pear, cochineal biogeographic prickly a native establish the to on attempt failed doddi a Cactoblastis and invasive Africa, fencing of South agent and agricultural control Australia In biological threatening 2009). a pest Carpenter, as important & an (Hight pear also production is prickly fruit it pear for where Moreover, Caribbean pest 2005). the important and where al., States an et range considered (Zapata distribution subsistence is native familiar its moth in The production provinces. fruit biogeographic Monte and Pampean 2014). two Carpenter, & Hight, cactorum Briano, Cactoblastis Logarzo, (Varone, area cultivated the semi in exotic households the this of In patches species However, cactus native 2014). two America, (Morrone, “card´on” landscape: South provinces the cactus southern in of biogeographic predominant ranges most Monte mountain the pre-Andean and are of different Chacoan slopes have eastern Prepuna, the (NWA) species Puna, area, Argentina northwestern These the of areas between 1985). are of limit (McFadyen, there However, the ranges Argentina are the inside use. of Eggsticks which plant burrow host areas in larvae of enemies. the biogeographic patterns and hatch, several natural ranges eggs in geographic of the found avoidance once cactus be the and a can plant in of cactus appearance role a the a of Cactoblastis mimicking ( play spine (eggstick), pears may the prickly hosts other of or that cactus the stems to areola feature of moths the top A these to of on attached adaptation one 1940). of stacked degree (Dodd, are The spine eggs resources. since feeding striking and breeding is groups 2015). as herbivorous Cactaceae Whiteman, to several of & compared use in Lapoint, herbivores Wiens, speciation specialist 2015; genus and al., herbivory in The shifts et between richness Simon host relationship species 2006; between a greater al., link Funk, of the et 2017; the (Funk hypothesis as al., and such the et taxa, abundant, of supporting (Forbes non-herbivore-related is sympatry evidence challenge diversity in Moreover, the insect species involves multiple 2006). and and often of Etges, parasitoid intra plants coexistence & modified the of and host Nosil, environments, enable basis mating new that new ecological of factors sources, ecological and interactions; food acquisition to new genetic lead environments, the can diverse the shifts chemically since Host facing of diversification 2005). study Dicke, and & the Loon, specialization van for Loon, Van models (Schoonhoven, divergence excellent interspecific are insects Phytophagous Cactoblastis Opuntia atbatsdoddi Cactoblastis ti obflta h netwas insect the that doubtful is it , Cactoblastis species, ohseis ic hne nde aebe eotda atrta rvsdvricto in diversification drives that factor a as reported been have diet in changes since species, moth Echinopsis nti aierne n opooia ieecsaepeetrgrigteegtc and eggstick the regarding present are differences morphological and range, native this in Vrn ta. 02 aoee l,2014). al., et Varone 2012; al., et (Varone rcoeesatacamensis Trichocereus pce.Isde nldssvrlntv pce ftegenus the of species native several includes diet Its species. a oelmtdgorpi itiuinadde rat.I a enrpre feeding reported been has It breadth. diet and distribution geographic limited more a has atbatscactorum Cactoblastis Cactoblastis a ie itbedhadamr xeddgorpi itiuinta h other the than distribution geographic extended more a and breadth diet wider a has Opuntia .ddi .cactorum C. doddi, C. ossso ebvru oh aiet otenSuhAeiaseilzdi the in specialized America South southern to native moths herbivorous of consists e.Although Lem. a eotdfeigon feeding reported was and .sulphurea O. Denmoza p. rclma at n tr ofe Vrn ta. 09.A es three least At 2019). al., et (Varone feed to start and cacti columnar or spp.) p.oelp(cayn 95.Teeaescnito rdfrssi the in forests arid of consist areas These 1985). (McFadyen, overlap spp. Opuntia Hirc,13;Mn,16) aveo atpii ohthat moth cactophilic a of Larvae 1969). Mann, 1939; (Heinrich, rdmnn atsseisi h ot,Pn n Prepuna and Puna Monte, the in species cactus predominant a , Cleistocactus pni sulphurea Opuntia var Berg, .cactorum C. pasacana or pni ficus-indica Opuntia p.ta eeipre ntemd10sfruea natural as use for mid-1700s the in imported were that spp. atbatsbucyrus Cactoblastis .bucyrus C. .ficus-indica, O. .bucyrus C. 2 .cactorum C. ee xR ex Weber .ficus-indica O. p a eotdb an(99 sahs of host a as (1969) Mann by reported was sp. .atacamensis T. atbatscactorum Cactoblastis ic l te ot of hosts other all since eercnl olce nteclma cactus columnar the on collected recently were ile xSl-ykadteebeai giant emblematic the and Salm-Dyck ex Gillies pce itiue rmcnrlwestern central from distributed species a , u h eod r eadda uncertain as regarded are records the but sa naieseisi ohteUnited the both in species invasive an is mlr( umpler ¨ ilrcnas efudaon some around found be also can Miller yr and Dyar, Opuntia rp r nesnilcmoetof component essential an are crops sacmo otof host common a as .atacamensis T. Opuntia p.boiest n prickly and biodiversity spp. .cactorum C. atbatsdoddi Cactoblastis a efudi Chacoan, in found be can .cactorum C. p.adteexotic the and spp. rmhereafter). from a introduced was .bucyrus C. r nthe in are Heinrich, C. Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. n usqetyba uie.Lbaiswr rcse ihIlmn Btfrcutrgnrto nthe on generation cluster for indexes cBot TruSeq Illumina introduce with that primers processed oligo were with Libraries Freedom, amplified purified. and Inc., was bead Instruments fraction purified pool, eluted subsequently (Sage was each gel instruments For and DNA The purified. 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To location survey. the per et genetic in our Varone collected in and were included larvae and (1985) instar McFadyen I-IV following each and moth, collected, Eggsticks to cactophilic related were assigned a were they of collected on which individuals individuals collected two on province field, collected plant we the Finally, host in (2014). the species al., these on genus distinguishing based the of of species difficulty species the as any In identified to NWA. for plants Due in reported of sampled numbers been populations equal not the on has to collected that relative individuals plant allopatric 10 largely included were we sites species collecting addition, these three of the locations of The populations of sympatric sulphurea populations O. of allopatric plants included Samples also host we range. the design region, on native this feeding the In species 1). in three quimilo (Figure diet of NWA each in their of obtained of populations were two part least as at reported from individuals 70 collected We extraction DNA and sampling role Taxon important contact. secondary an ongoing after played of even plants hypothesis species host Methods across our that flow however, Our suggesting gene species, species. rejected, limiting between between divergence was and occurring during populations within flow sympatric admixture gene in genetic historical hybridization and diversity revealed genetic data interspecific genomic and and sympatric We intra of of pairs data. each included patterns mitochondrial We of on populations. and populations feeding between distant SNPs flow population of geographically gene genome-wide moth interspecific use the contemporary of host and of combination historical plant status a of the using evaluated patterns moths also and cactophilic distribution related biogeographic closely regarding features uncertain Cactoblastis are there Since 2006). al., et (Funk Lepidoptera ° )adhmdt 7 ) n fe acigte eerae ni hyrahdtefut instar fourth the reached they until reared were they hatching after and %), (70 humidity and C) Schum, nMnoaPoic and Province Mendoza in p. eivsiae hlgorpi atrsadpyoeei eainhp nti roof trio this in relationships phylogenetic and patterns phylogeographic investigated we spp., .doddi C. pni anacantha Opuntia on .sulphurea O. pni megapotamica Opuntia and , pg ob sda noutgroup an as used be to Speg. Cactoblastis .bucyrus C. .doddi C. .cactorum C. NspI 3 p.Ti apigshm loe st investigate to us allowed scheme sampling This spp. and and on .cactorum C. .baumannii C. MboI .atacamensis T. Cactoblastis rca.i aPmaPoic,respectively. Province, Pampa La in Arechav. a olce on collected was ° ni N extraction. DNA until C etito nye eeslce sthe as selected were enzymes restriction . rmcnrlAgniafeigon feeding Argentina central from nSnig e seoProvince. Estero del Santiago in n erhdfrfopit of footprints for searched and , Tbe1) (Table Tucumania .ficus-indica O. .baumannii, C. . norexperimental our In p rmFormosa from sp. < 0m fclean of mg 30 and Opuntia host a Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. atrso ula iest o ahpplto eecaatrzduigosre ( criterion. observed cross-entropy using the characterized using were assessed population was ( each value erozygosities for K 2015). diversity optimal Francois, nuclear The populations, & intra-and of allopatric K. (Frichot and Patterns of of package sympatric value for proportions LEA each 8 to for estimate the 2 repetitions from using to 100 ranging values R with species, K ancestral in for recognized calculated This implemented K were the coefficients was from frequencies. 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Data may be preliminary. upeetr al .W sindt 1adP h ypti ouain fteseistse (A-B: tested species the of D populations P2. sympatric or (D the P1 P3 P3 and and and P3 P1 P1 between between to 0) flow bucyrus assigned = gene We C. (D indicated The introgression 4. 3) 2020). Table (D no Overcast, Supplementary P3 of [?] 25%) & and hypothesis (Z-score (msl= (Eaton P2 null zero dataset Eaton between the from by largest test different recommended the to significantly as used values used available the we was test data used to method statistic We of 2011) sympatry. 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Estero unlinked 1688 del of matrix a in resulting .ccou,C doddi C. cactorum, C. eioismelpomene Heliconius and .doddi C. .doddi C. > C-D: ; and ) epromdAB/AAtsigacrigt h ceedlnae in delineated scheme the to according testing ABBA/BABA performed We 0). .bucyrus C. and .cactorum C. Kihlye l,21)(eiotr:Nmhlde.Atraiemodels Alternative Nymphalidae). (Lepidoptera: 2015) al., et (Keightley .bucyrus C. n he needn nrseicslt eaaigbt popu- both separating splits intraspecific independent three and , and a atal vrapn egahcrne,introgressive ranges, geographic overlapping partially had π .02 a siae sn ain n nain sites invariant and variant using estimated was 0.0012) = .doddi C. 5 E-F: ; https://github.com/isaacovercast/easySFS .bucyrus C. μ .cactorum C. f29 10 x 2.90 of ) and Δ )acrigt h formula: the to according i) .cactorum C. olwdb second a by followed -9 siae o the for estimated ,ada P2 as and ), < )or 0) π/ 4 μ ) Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. o dA aalbea CIa iPoetPJA673.Teaeaenme fpie-n ed per reads paired-end of number average The untangle PRJNA666743). to 1.7x10 BioProject and was as flow, NCBI individual gene at of (available patterns ddRAD diversity, for and in structure relationships genetic phylogenetic the interspecific and intra data investigate To mtDNA and ddRAD assembly, genome Draft the (Excoffier model of Results intervals best the confidence from 95% values initialized 2013). construct using al., to replicates et used bootstrap was 100 approach running parameters bootstrapping estimated parametric a Finally, model. lseigaayi lorvae o u oiebesgaso ditr nsmarcppltoso the of populations sympatric in doddi admixture C. of effect, doddi signals In noticeable C. 2b). but (Figure low species revealed three also analysis Clustering bucyrus. 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I au f0adtehgetACweight AIC highest the and 0 of value AIC .baumannii C. niiul hrdvrat ihsympatric with variants shared individuals aalbea ebn W625M041)(Supplementary MW064275-MW064313) Genbank at (available ) .cactorum C. oh,attlo 0seiesfo 2lctoswr genotyped were locations 12 from specimens 70 of total a moths, ouain .. o neietcretcnatwt closely with contact current evident in not i.e., population, niiul rmNAsoe eei ditr ihboth with admixture genetic showed NWA from individuals niiul rmNAsae ainswt sympatric with variants shared NWA from individuals 6 rmhratr ept ti otaitr ihthe with contradictory is it despite hereafter, from sn oa f2.8x10 of total a using rmNWA from .doddi C. .Tu,w rsne h eut obtained results the presented we Thus, ). ouain eercvrda separate as recovered were populations .ddi .cactorum C. doddi, C. .bucyrus C. .baumannii C. 5 ed aeaera length= read (average reads notosprtdclusters separated two into .cactorum C. to ω .cactorum C. evda h best the as served i .bucyrus C. and .cactorum C. .bucyrus C. appeared cluster, and C. < a Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. dA aa sn naeaeo 287ulne dA Ns a sdt erhfrfopit of footprints modelling for Demographic search of to populations tests sympatric used These between was hybridization and 4). introgressive SNPs, Table of (Supplementary ddRAD hypothesis and test unlinked the ABBA/BABA to 12,897 four-taxon support of gave the average with hybridization an introgressive using data, 3a). (Figure ddRAD Mya) the between 2.24 that split - 1.35 Mya), the = 4.08 that interval - and 2.78 Mya), = 2.84 interval genus credibility the of that doddi suggested (95% data (Mya) on mtDNA ago collected on moths based of times subclade divergence other the on and (NWA) exclusively feeding on collected individuals bucyrus specimens Likewise, encompassed included subclade subclade Pampa. 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Cactoblastis st .bucyrus C. , .bucyrus C. .3 o ohdRDadmtDNA). and ddRAD both for 0.939 = a h ihs altp iest as diversity haplotype highest the had eegntclymr ies hnthe than diverse more genetically were rmSnig e seo siae of Estimates Estero. del Santiago from ouain perda differentiated as appeared populations curd17 y 9%o credibility of (95% Mya 1.79 occurred .doddi C. .doddi C. Opuntia st Fgr a.O h te hand, other the On 3a). (Figure eemr ies hnNWA than diverse more were a bevdbtensympatric between observed was and Fgr c.Itrsigy two Interestingly, 3c). (Figure .ficus-indica O. niiul,csigduton doubt casting individuals, and rgntd34 ilo years million 3.41 originated priori a rmSnig e Estero. del Santiago from feeders .doddi C. .bucyrus C. .sulphurea, O. .cactorum C. .bucyrus C. lne,bcueof because planned, .cactorum C. ntemidpoint the in .atacamensis T. lo the Also, . eddnot did We . .cactorum C. .bucyrus C. appeared n the and and to , C. C. C. C. Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. ieetae is a cainlyhbiiei auei ypty(isne l,22) iial,a Similarly, 2020). al., et genus (Giesen the sympatry of in butterflies nature of in study hybridize isolation genetic that occasionally reproductive suggest pioneering a may of studies to evolution genetic flies, points the Likewise, differentiated study by populations. This followed sympatric 2020). contact cynipsea among al., Sepsis secondary hybridization et after prevents (Tonzo overlap currently flow distributions that gene their been historical where also of area has scenario the hybridization Zhang, in introgressive 2018; introgression of grasshoppers Simon, past Evidence & the et (Ward Tonzo 2016). species in 2014; Kronforst, related al., reported distantly & et and (Clarkson Moreira, allopatric sympatry Mallet, between in frequently Dasmahapatra, occurring less generally and species, 2020), insect al., several in reported been have between analysis events clustering hybridization between and introgressive tests and suggested NWA, ABBA-BABA coalescent-based results of by These results supported the sNMF. divergence with using intraspecific concordant before are overlapping Fastsimcoal2 and with using after species simulations admixture 2019). related historical al., closely of et of footprint Poveda-Martinez pairs The 2014; in even al., barrier flow, et isolating gene (Clarkson an to distributions barrier as biogeographic effective role an These important as an populations. documented played allopatric been differentiated plants presently host clearly between alternative and were to sympatry among species adaptation of related that areas included in suggested closely admixture that results these of moths that evidence economies. of revealed the agricultural-based group despite analyses local a phylogenetic affecting of negatively and including biodiversity, history Clustering approach cactus evolutionary threaten genetic the that population on species light a pest two shed combining that study ( hypothesis Our mtDNA phylogeographic genus and the divergence. of (ddRAD) moths intraspecific nuclear cactophilic after American South and the before in variation genetic interspecific Cactoblastis and intra of survey The interspecific of Discussion episode of recent most populations The sympatric intervals). between confidence bucyrus for occurred 3 Table flow (see gene 4) (Figure which ago in years populations thousand sympatric between three occurred flow the gene involving between flow obtained model was gene likely flow most historical doddi gene the assumed of species rate model within highest divergence This the after flow 5. gene model interspecific was of scenarios considered we When ago. of years populations 41,641 estimated occurred conspecific we 2019), presumptive al., the et Varone within 2014; by al., while shared et Mya, ancestor (Varone 2.22 common species last native the in occurred year that flow per gene ( generations of two populations rate Assuming highest sympatric the between of divergence that flow before pairs revealed gene flow species) gene historical within (assuming only simulations historical populations between considering These sympatric assuming scenarios models 5). and one Table to tested allopatric was Supplementary compared we of 4; model as (Figure When likely species species, most of three 2). the of pairs the divergence, Figure scenarios involving intraspecific (Supplementary alternative flow before divergence gene depicting flow intraspecific gene models after interspecific tested and of simulations before These three flow, the Fastsimcoal2. gene of using history simulations demographic based population and Species and Cactoblastis uigapro ftm ewe . huadt 32tosn er ago. years thousand 13.2 to thousand 1.8 between time of period a during .bucyrus C. .bucyrus C. eeldacmlxeouinr itr novn itrcleioe fgntcinterchange genetic of episodes historical involving history evolutionary complex a revealed .and L. .doddi C. .cactorum C. pce ntefc fitrpcfi eeflw iia neths ln soitoshave associations plant insect/host Similar flow. gene interspecific of face the in species and m .00) olsetsmltosas eeldta h otacetinterspecific ancient most the that revealed also simulations Coalescent 0.00108). (m= essnecynipsea Sepsis and .doddi C. .bucyrus C. .bucyrus C. mcsu minutissimus Omocestus and .cactorum C. COI mgainrt e eeain =004)a oprdwt h other the with compared as 0.0049) m= generation: per rate (migration .bucyrus C. and akr ihsreso otpatuealwdu opooea propose to us allowed use plant host of surveys with markers ) iegdmr eety 0,3 er.Smlraaye hwdthat showed analyses Similar years. 700,339 recently, more diverged eadr&Sue Dpea esde,torltdadgenetically and related two Sepsidae), (Diptera: Spuler & Melander .cactorum C. .doddi C. and rmSnig e seo cnro fhsoia eeflow gene historical of Scenarios Estero. del Santiago from .cactorum C. .bucyrus C. .doddi C. uigasotpro ftm rm1. huadt 16.1 to thousand 12.9 from time of period short a during Heliconius 8 n h clade the and Brull´e and .cactorum C. Cactoblastis or and NAadSnig e seo divergence Estero) del Santiago and (NWA aril nldn ar fsmarcand sympatric of pairs including Latreille .cactorum C. .bucyrus C. mcsu uhagonii Omocestus .doddi C. and p.wsasse sn coalescent- using assessed was spp. .doddi C. + and m .05) olwdby followed 0.00356), (m= .doddi C. .bucyrus C. and , .bucyrus C. oia suggesting Bolivar .cactorum C. and curdabout occurred .bucyrus C. Tbe3). (Table ) - C. C. in Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. aieylwgntcdffrnito ugse atr fetniegn o ntedsrbto ag of range distribution the in flow gene extensive of pattern a suggested differentiation genetic low latively within variation, was intraspecific evidence to available within corresponded The differentiation structure cacti. genomic columnar in of on would pattern occurred breeding the isolation as moths whether postzygotic these define of for to plant status sufficient testing host not taxonomic experiments isolation, the breeding prezygotic confirm evaluate and to to help experiments, aimed performance experiments choice 2020; and mate al., preference Moreover, et genome. nuclear Hood the ex- 2017; of niche species al., a a either et merely or on (Forbes represented host feeding observations differentiation new present in suggesting a our role km), to if a (˜500 determine pansion played to apart Thus, have far that 2020). may were al., Poveda-Mart´ınez hypothesized distance populations et We by these However, 1985). isolation populations. McFadyen, that moth 2021; between al., divergence from et tic shift values (Barbarich plant the genera host than on different the mtDNA collected for of moth greater although times allopatric Cactoideae, 1.1 the and and 1.2 (NWA) and stricto markers nuclear for within greater estimated The times 1.5 2014). and 2.5 al., were et within Varone differentiation either 1985; greater of revealed McFadyen, markers 1969; genetic recognized both Mann, three 1939; the Heinrich to ding 2021; compared al., when cactus preferences et columnar ecological the and on ferences hypotheses. collected the these moths introgression, test of cactophagous to extent The and segments direction introgressed the of determine function to and necessary distribution, are analyses comparative of genome whole capabilities between the large flow of of gene cultivated evolution introduction interspecific the the favoring after zone in particularly contact role tests, a introgression be by may revealed indica and as crops species, pear related prickly these of pest important an putative cactorum of ancient the populations of of maximum. pairs distant glacial product distribution the between last a for flow the as during gene (2020) and suggested occurred interspecific al probably was revealed et that flow analyses Tonzo flow Introgression gene gene by interspecific recent reported of where were not early grasshoppers, populations and results during sympatric of introgression between Similar occurred variants species probably shared populations. gene sympatric recent of that allopatric of of excess flow pattern to an common gene and A compared differentiation ongoing completed. not as reduced or was in isolation recent result seemed reproductive than flow should which in gene flow rather shift of than ancient plant footprints sympatry host of These of of mtDNA. stages product and areas the ddRAD in for be lower either was to populations, species allopatric of between pairs differentiation between genomic overall populations, genetic allopatric in for admixture 2018). testing though Simon, Even approaches & complementary (Ward crosses However, laboratory years. experimental the 125 of in contrast, over isolation In reproductive for 2013). supported sympatric al., admixture et been indicating (Martin have sympatry, species in Hebert, related variation closely shared these increased between moth of flow trend gene genome-wide interspecific pervasive a revealed populations, allopatric Cactoblastis .bucyrus C. ltlaxylostella Plutella .bucyrus C. lnain.Tu,w a s,wehritrpcfi eeflwbtenseismyhv lyda played have may species between flow gene interspecific whether ask, may we Thus, plantations. .cactorum C. nticue nti td)hv enrpre Vrn ta. 04.I hsregion, this In 2014). al., et (Varone reported been have study) this in included (not .ficus-indica O. .doddi C. Snig e seo.Hwvr ti motn ont hti h oteneg fthe of edge southern the in that note to important is it However, Estero). del (Santiago .cactorum C. .atacamensis T. p a sindto assigned was sp. .cactorum C. and or .doddi C. and . nitoue eti utai,adtenative the and Australia, in pest introduced an L., .doddi C. .baumannii C. n h mlmtcntoa monument national emblematic the and Cactoblastis .cactorum C. .bucyrus C. ihbt ula n iohnra akr,dffrnito a oe than lower was differentiation markers, mitochondrial and nuclear both with or ept h ogdsac ˜,5 m eaaigteeppltos hsre- This populations. these separating km) (˜1,350 distance long the despite nfc,F fact, In . or .doddi C. nsaunascendi statu in .baumannii C. .bucyrus C. to p.wr eetdbtenpeetysmarcadas between also and sympatric presently between detected were spp. ouainlvn on living population eetees ept u uvyrvae infiatpopulation significant revealed survey our despite Nevertheless, . .atacamensis T. .cactorum C. st epciey(upeetr al ) Both 3). Table (Supplementary respectively , ausbetween values Plutella o vnohrclma at)adalre representation larger a and cacti) columnar other even (or ybt lseigadpyoeei nlss However, analyses. phylogenetic and clustering both by eursadtoa olcin falpti populations allopatric of collections additional requires 9 .baumannii C. pce nAsrla ept hi blt ohybridize to ability their despite Australia, in species and rvc es,wsa motn rvro gene- of driver important an was versa, vice or , .baumannii C. .bucyrus C. .bucyrus C. .bucyrus C. .baumannii C. .atacamensis T. e nclma at ftesubfamily the of cacti columnar on fed hnbtenalpti populations allopatric between than n the and obcm motn et fthe of pests important become to aeaprn opooia dif- morphological apparent have ltlaaustraliana Plutella Cactoblastis oxsigppltosof populations coexisting , .baumannii C. epciey However, respectively. , .cactorum C. .baumannii C. pce (Barbarich species .bucyrus C. .cactorum C. edn moths feeding ady& Landry .ficus- O. (NWA) sensu fee- C. C. is Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. wre oGL) ...adLM r eiinso h coasisaaddb OIE n ANPCyT, and CONICET by awarded scholarships PhD of Tecnol´ogica Investigaci´on recipients (PICT1447/2016 Cient´ıfica are USDA y la L.M. from para and obtained D.P.M. Fondo C´omputos was G.L.). de Funding and de to resources. awarded Centro 10201, computational the Recursos Bill of and y Farm Bioc´odices use SA, Ambiente Fuego, granting APHIS-PPQ, Polares, del for Ciencias Tierra members (CeCAR) de de to Rendimiento support. Instituto gratitude Nacional Alto the fieldwork our Universidad thank express for the We and of comments. Guala analyses, Naturales insightful Mariel data for with and Tonzo Lab Vanina Evolution assistance, of of technical assistance the for appreciate Restrepo also We Sanchez Andres appreciate We invasive resistance the insecticide between pyrethroid flow for gene Acknowledgements acquired sense, introgression this widespread 2020). In a., and question. et hybridization (Valencia-Montoya open of an evidence still with between is hybridization area armigera interspecific invaded Helicoverpa via its variants in genetic species of invasive distantly acquisition in the reported Whether cactorum been 2009). Carpenter, also have & threatening 2016). introgression 1989, al., of since patterns et America but (Zhang Lepidoptera species, related related of closely trio in the hybridization in (Fagua flow Lepidoptera gene several cactorum interspecific of including of trio taxa, evidence this many the The with of of coincident 2020). divergence origin period Hewitt, the the time 2019; in that This al., resulted Pleistocene. suggested et early refugia markers in the glacial genetic frequently the and of of Pliocene both been occurrence estimations Upper has on mentioned, the it based between incongruence as occurred the species species Despite (ILS), between as 2019). sorting divergence considered al., lineage of typically et incomplete time (Moreyra are or taxa incongruences hybridization insect analyses, mito-nuclear diverged past coalescent-based recently Such either and flow. of ABBA-BABA gene consequence tests, a interspecific introgression experienced both having in as identified also were trees, incongruence species tree, the mtDNA the and in ddRAD, the In congruent. on feeding moths and studied for the one for now. inferences to phylogenetic up based (mtDNA) registered tis of mitochondrial been and presence has (ddRAD) the nuclear what Both of than confirmation common The more NWA. be in could plantings association ornamental as used doddi mainly and that uncommon indicating and observations previous confirmed cactus different results exotic our the However, includes 1979). also demographic al., the et that understand Zimmermann pest. to documented cactus necessary this are reports in species Previous flow the Varone gene of & of distribution Corona dynamics (Fuentes wider isolation and a over postzygotic history plants or host sexual more exotic of including and evidence studies any Further show comm). not area did personal southernmost Our km the 1,000 2011). of than al., representative more population et a (Marsico include diversity to by mitochondrial area inhabited high sampling identified the extended distribution scheme species’ sampling the of edge mitochondrial southern the using survey previous a cactorum ouain eoee h he xetdcae codn otepplto eei uvy(iue2-C); (Figure survey genetic population the to according clades expected three the recovered populations edn on feeding .cactorum C. .ficus-indica O. eotdt nld eea igorpi rvne nAgnia(aoee l,21) Moreover, 2014). al., et (Varone Argentina in provinces biogeographic several include to reported , n lsl eae pce a lydarl nfeigaatto n dpierdaino this of radiation adaptive and adaptation fueling in role a played has species related closely and ohbiiewt te pce nisntv ag.W on vdneo nin introgressive ancient of evidence found We range. native its in species other with hybridize to .cactorum C. .ficus-indica O. .bucyrus C. nte for another , unradtenative the and Hubner .baumannii C. ace takdby attacked patches nAgnia nebedn xeiet novn ohppltossprtdby separated populations moth involving experiments Interbreeding Argentina. in .ficus-indica O. perda h xenlcae hs pce novdi h mito-nuclear the in involved species These clade. external the as appeared Opuntia eie rmteueo eei akr rpssta hsinsect-plant this that proposes markers genetic of use the from derived .doddi C. .doddi C. oee,temtcodiladtencerteswr o entirely not were trees nuclear the and mitochondrial the However, . p.boiest n rcl erarclua rdcin(Hight production agricultural pear prickly and biodiversity spp. COI atbatscactorum Cactoblastis n h hr included third the and .doddi C. eioep zea Helicoverpa srsrce to restricted is nisde MFdn 90 cayn 95.W on two found We 1985). McFadyen, 1980; (McFyden, diet its in eeicuigasto ouain htddntcmrs the comprise not did that populations of set a including gene .cactorum C. .cactorum C. 10 Cactoblastis nubnlctoswhere locations urban in .sulphurea O. ouain soitdwt diinlnative additional with associated populations odei neapeo w oh species moths two of example an is Boddie perda h otetra ld,while clade, external most the as appeared pce eosrtdtecpblt of capability the demonstrated species a enatcigntv at nNorth in cacti native attacking been has .bucyrus C. Mn,16;MFde,1985; McFadyen, 1969; (Mann, olce on collected .ficus-indica O. .atacamensis T. a scarce was Cactoblas- .doddi C. C. C. C. Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. rco,E,Mtiu . rulo,T,Buhr,G,&Fa¸os .(04.Fs n ffiin siainof estimation efficient and Fast studies. (2014). Fran¸cois, association O. & ecological G., coefficients. Bouchard, ancestry and T., individual Trouillon, landscape F., for Mathieu, le- package E., Frichot, High R (2004). Evolution An and R. LEA: Ecology Dallai, (2015). in & Methods Fran¸cois, O. V., & Scali, E., Wilson, V., Frichot, of & Paola, populations A., non-infected De and H. M., Wolbachia-infected Isotomidae). Pellecchia, Widmayer, between (Collembola, P., differentiation K., speciation. P. genetic insect A. Fanciulli, of initiating Ward, vels I., in S., Negri, shifts E. F., host Frati, Tvedte, of C., role A. particular the Hippee, 1126-1137. Revisiting N., Sperling, (2017). S. & J. Devine, . C. . . A., J., A. T. of Simonsen, Forbes, diversification J., evidence. Prins, and molecular De biogeography demographic on K., delimitation, B. based Robust Genus Tortricidae) Byun, (2013). J., (2019). J. 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(8), oeua ilg n vlto,29 Evolution, and Biology Molecular oeua ilg n Evolution and Biology Molecular aueCommunications Nature Pedobiologia 8,1917-1932. (8), het . aga,T,W,C . i,D,...&Dumn,A .(2014). J. A. Drummond, & . . . D., Xie, H., C. Wu, T., Vaughan, D., uhnert, ¨ Genetics , , 6 58 8,925-929. (8), rcoeespasacana Trichocereus LSGntc,9 Genetics, PLoS 34,23-32. (3-4), Anopheles , 48 , https://doi.org/10.1093/molbev/mss086 ytmtcEtmlg,44 Entomology, Systematic 196 56,461-468. (5-6), 4,973-983. (4), , 5 https://doi.org/10.1111/2041 iln pce lmntsamjrgnmciln u not but island genomic major a eliminates species sibling 1,1-10. (1), 11 8,1969-1973. (8), , , 11 28 1) e1003905. (10), 1,122-137. (1), 8,2239-2252. (8), https://doi.org/10.1534/genetics.113.160572 https://doi.org/10.1038/ncomms5248 nl rpn juje˜na (Argentina). Prepuna la en https://doi.org/10.1093/molbev/mss075 LSCmuainlBiology Computational PLoS 1,19-38. (1), hrsoer Lederer Choristoneura atbatsbucyrus Cactoblastis clgclIndicators Ecological Evolution osmacandida Folsomia (Lepidoptera: (Lepidoptera: eit ela de Revista Molecular , , 10 71 Bioin- 121, , (4), (5), Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. fseisfrbooia oto fpikypas( pears prickly of control 159-168. biological for species of characteristics of Larval (1985). E. R. McFadyen, Jiggins, Australia. & in [Cactaceae] . . (1980). . F., R.E. Simpson, McFadyen, R., in (2011). flow J. E. gene Walters, with C., M. speciation Salazar, for Welch, J., evidence , N. Genome-wide & Nadeau, (2013). E., K., D. K. C. J. Dasmahapatra, H., McClure, S. populations. P., Martin, invasive C. for introductions Brooks, multiple of N., and range 13 invasion G. native of the Ervin, from history diversity E., documented genetic L. of patterns Wallace, Geographic D., T. Marsico, mites. and insects complexity. Cactus-feeding genome 158 of (1969). origins The J. (2003). Mann, S. J. Conery, & tree, M., construction. Short network Lynch, haplotype for 6 (2015). software full-feature Evolution, A. Popart: and phylogenies. Stamatakis, Ecology SNP (2015). & in D. inferring Bryant, M., & for W., N. corrections J. Leigh, bias A. acquisition Oca, new De tree: J., 64 wrong Felsenstein, biology, tree, L., right B. tree, Banbury, Jiggins, long & D., A. . . . Leache, J., in Mallet, rate mutation K., spontaneous K. the J. 32 Dasmahapatra, of F., Evolution, Feder, Estimation Simpson, W., & (2015). R. D. Ness, C. package A., errors. R . Pinharanda, . associated An D., . their P. diveRsity: Keightley, and L., (2013). parameters A. W. genetics P. population human- Yee, Prodohl, Evolution of & recent and M., exploration W., Ecology a W. and Glover, following Crozier, estimation F., B., the isolation T. for reproductive S. Cross, P., drive McGinnity, Sim, K., shifts fly, Keenan, M., maggot plant apple M. host (Lepidoptera: the ages. repeatable cactorum Doellman, of and introduction Cactoblastis H., mediated ice Rapid male T. of Powell, (2020). Quaternary phenology L. R., Flight States. the G. United southeastern (2009). Hood, of the E. in J. legacy latitudes different Carpenter, at genetic & Pyralidae) American D., The the S. Hight, of revision a (2000). toward 913. contribution G. a Hewitt, Phycitinae: Phycitidae. family cactus-feeding the The of moths pyralidoid (1939). C. F-statistics. hierarchical Heinrich, test and & compute of to R . species . 5 for . package related Notes, a B., closely Hierfstat, Misof, two (2005). between R., J. Goudet, reinforcement Shimizu-Inatsugi, at and Preprint K., bioRxiv. admixture K. flies. of Shimizu, sepsid signals A., European Genomic M. (2020). Schaefer, M. U., Kapun, W. Blanckenhorn, A., Giesen, associations positive consistently exhibits divergence taxa. Ecological 3209-3213. disparate (2006). across J. isolation W. reproductive Etges, & with P., Nosil, J., D. Funk, 23 4,857-868. (4), https://doi.org/10.5479/si.03629236.256.1 https://doi.org/10.1038/35016000 1) 1817–1828. (11), 1,184-186. (1), https://doi.org/10.1073/pnas.0508653103 6,1032-1047. (6), 1,239-243. (1), https://doi.org/10.1101/gr.159426.113 https://doi.org/10.1111/j.1471 , 4 noohg,25 Entomophaga, Cactoblastis 8,782-788. (8), 9,1110-1116. (9), Lp:Pyiia]frtebooia oto of control biological the for Phycitidae] [Lep.: https://doi.org/10.1101/2020.03.11.985903 rceig fteUie ttsNtoa Museum National States United the of Proceedings 1,37-42. (1), hgltspomonella Rhagoletis Opuntia Cactoblastis rceig fteNtoa cdm fSciences of Academy National the of Proceedings 12 https://doi.org/10.1007/BF02377520 ultno h ntdSae ainlMuseum. National States United the of Bulletin spp.). p.Lpdpea yaia)adteselection the and Pyralidae) spp.(Lepidoptera: eioismloee oeua ilg and Biology Molecular melpomene. Heliconius ultno noooia eerh 75 Research, Entomological of Bulletin atbatscactorum Cactoblastis lrd Entomologist Florida Heliconius . Science Evolution , aue 405 Nature, butterflies. 302 , 74 54) 1401-1404. (5649), 1,156-168. (1), ilgclInvasions Biological roeesmartinii Eriocereus Br)spotthe support (Berg) oeua Ecology Molecular , eoeResearch Genome 92 2,208-216. (2), 68) 907- (6789), . ehd in Methods Systematic , Methods 103 (1), (9), 1– , Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. oeei tlt fmtcodilgn eune n oplto fcnevdplmrs hi reaction chain polymerase phy- conserved and York: of weighting, Evolution, compilation New a (1994). P. and biology. Flook, sequences Insect-plant & gene H., mitochondrial Liu, (2005). Ge- of B., M. Crespi, utility Dicke, A., logenetic Beckenbach, J., (2007). F., Frati, J. C., C. Simon, Loon, the van Kerdelhue, B., of & Press. Loon, University population Van M., Oxford atypical M., Branco, phenologically L. Schoonhoven, R, a of M. differentiation Paiva, allochronic 941. moth. E., processionary time: Magnoux, pine through J., sets. isolation data Rousselet, netic large of H., & analysis E., Santos, polymorphism S. sequence Ramos-Onsins, 34 P., DNA Huelsenbeck, Evolution, Librado, 6: & and S., DnaSP Biology Guirao-Rico, model C., large . (2017). . J. a . A. 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Laprida, Morrone, F., the Almeida, of history J., evolutionary 14 Hurtado, the ONE, about J., us tell Mensch, mitogenomics N., N. Moreyra, https://doi.org/10.1098/rspb.2006.3767 http://dx.doi.org/10.11646/zootaxa.3782.1.1 , ytmtcBooy 61 Biology, Systematic 7 https://doi.org/10.1371/journal.pone.0220366 5,e37135. (5), 1) e0220676. (11), oh:a nertv approach. integrative an moths: https://doi.org/10.1002/ece3.6702 75,1034-1044. 97(5), Hypogeococcus pungens 1) 3299-3302. (12), rceig fteRylSceyB ilgclSine,274 Sciences, Biological B: Society Royal the of Proceedings 3,539-542. (3), . iSine 57 BioScience, oeua ilg n vlto,27 Evolution, and Biology Molecular xodSre nEooyadEouin e ok Oxford York: New Evolution. and Ecology in Series Oxford oeua Ecology, Molecular pce ope Hmpea suoocde o Argentina, for Pseudococcidae) (Hemiptera: complex species 13 8,699-705. (8), rspiabuzzatii Drosophila Cactoblastis Heliconius 61) 3617-3633. 16(17), butterflies. at itrclcaictoso a on clarifications historical past: clg n vlto,10 Evolution, and Ecology naso h Entomological the of Annals lse rpeagroup)?. (repleta cluster 7,1659-1672. (7), Zootaxa LSGntc,8 Genetics, PLoS lSOE 14 ONE, PloS , 11) 935- (1612), 3782( Molecular ) 1- 1), (19), PloS PloS (7), (6), Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. a ed r vial tNB sBorjc PRJNA666743. BioProject as Fastsim- NCBI with at tested available are models reads demographic Raw and files, Figshare: input in study, available current are the coal2 during insects generated cactus-feeding datasets some The of list Annotated Genome-wide (1979). Availability E. (2016). Data R. R. McFadyen, M. & America. E., Kronforst, South H. & of Erb, R., G., G. H. Moreira, Zimmermann, J., Mallet, related distantly K., del among K. introgression sustentable Tecnologia Dasmahapatra, Manejo Ciencia, W., de Zhang, (2005). en Ministerio integral del M. Ndeg2. ( productivo Productiva Cartilla Karlin, tuna Innovacion Productiva, polo la & Innovacion de un de Federales e O., de procesamiento Proyectos R. Desarrollo y organica. Proyecto cultivo Coirini, certificacion tunales, Cordoba: F., los de J. Lucero, norte de Kitano, el Manejo O., & Grandes, U. Salinas Pungitius . Karlin, . ecosistema between . M., contact Y., R. secondary Machida, Zapata, after J., introgression A. and Nagano, divergence insect A., of across Toyoda, sticklebacks. patterns diversification H., Takahashi, Genome-wide increases R., Herbivory (2020). Kakioka, (2015). Y., K. Y. N. structure. Yamasaki, Whiteman, population cryptic & of T., analysis between R. the clades. Lapoint, admixture for J., F-statistics genomic J. 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Valencia-Montoya, distributions. of affair: overlapping evolution old partly an subsequent with of the grasshoppers footprints 2254-2268. diverged and Genomic recently hybridization two (2020). historical in J. (2015). reveal Ortego, R. data Streiff, & polymorphism & A., Papadopoulou, . . V., . Tonzo, P., insects. Nouhaud, herbivorous J., in Jaquiery, host-plants E., to Jacquin-Joly, 413-423. E., adaptation Guy, of E., Genomics d’Alencon, C., J. Simon, primers. https://doi.org/10.1093/gbe/evy224 aueCommunications Nature naso h noooia oit fAmerica of Society entomological the of Annals 1358-1370. , https://doi.org/10.1007/s10530-014-0670-9 eioep armigera Helicoverpa hlspia rnatoso h oa oit ,375 B, Society Royal the of Transactions Philosophical atbatsCactorum Cactoblastis caZooiaLilloana Zoologica Acta tp:/o.r/0j nmn6084/m9.figshare.13118249.v1 https://doi.org/10.jh lrd Entomologist Florida , 6 moths. 1,1-7. (1), Heliconius Opuntia Lpdpea yaia)o ot n ot mrcnOpuntia American North and South on Pyralidae) (Lepidoptera: oeua ilg n Evolution and Biology Molecular , 33 2,101-112. (2), utryspecies. butterfly otseisi Argentina. in species host atbatscactorum Cactoblastis eoeBooyadEvolution and Biology Genome 14 1163-1173. , , 87 pni ficus-indica Opuntia 6,651-701. (6), rensi ucinlGnmc,14 Genomics, Functional in Briefings eoeBiology Genome 10) 20190548. (1806), ntentv range. native the in , ilgclInvasions Biological 37 oeua Ecology Molecular seisaoaia bajo aromaticas especies y ) 9,2568-2583. (9), , 17 atbatscactorum Cactoblastis , 1,1-15. (1), 10 eioep zea Helicoverpa iCnrl 64 BioControl, 1) 2973- (11), , , 16 29 Plutella (11), (12), (6), Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. einHs lnsmDAmDAmDAmDAdRDdRDdRDddRAD ddRAD ddRAD ddRAD doddi Cactoblastis mtDNA doddi Cactoblastis bucyrus Cactoblastis doddi Cactoblastis bucyrus Cactoblastis mtDNA doddi bucyrus Cactoblastis Cactoblastis bucyrus Cactoblastis doddi Cactoblastis bucyrus Cactoblastis mtDNA doddi bucyrus Cactoblastis Cactoblastis cactorum bucyrus Cactoblastis Cactoblastis doddi Cactoblastis cactorum Cactoblastis bucyrus Cactoblastis cactorum Cactoblastis mtDNA doddi Cactoblastis cactorum Cactoblastis cactorum Cactoblastis Estero del bucyrus Santiago Cactoblastis cactorum Cactoblastis NWA cactorum bucyrus Cactoblastis Cactoblastis doddi Cactoblastis cactorum Mendoza Cactoblastis plants Host NWA doddi Cactoblastis Pampa La cactorum Cactoblastis NWA cactorum Cactoblastis Region of population 2. Table collected. were moths which from plants of number the indicate parenthesis 1. Table interest. of conflicts no Tables have they that declare authors the The analy- drafted data E.H. and and D.P.M., File work manuscript. Supplementary the collections. molecular of field version conducted performed final D.P.M. Information the G.L. Additional to and contributions work. S.H., made the coauthors E.H., All M.F., designed manuscript. L.V., E.H. D.P.M., and ses. G.L., L.V., D.P.M., MW064275-MW064313. numbers accession contributions under Author Genbank at available CactoFuEDEI are haplotypes as Mitochondrial available is genome JADGIL010000000. draft reference The aaPoic aiueLniueHs ln species plant Host sp. Tucumania Longitude Cactoblastis Latitude bucyrus C. doddi C. doddi C. doddi C. Province doddi C. doddi C. cactorum C. cactorum C. cactorum C. cactorum C. cactorum C. cactorum C. cactorum C. Taxa ao apigof sampling Taxon esrso eei iest nsmarc(W)adalpti ouain ape o each for sampled populations allopatric and (NWA) sympatric in diversity genetic of Measures Cactoblastis on Cleistocactus .baumannii C. atacamensis Trichocereus sulphurea Opuntia ficus-indica O. sulphurea, O. megapotamica Opuntia quimilo O. O.ficus-indica pbsdo tN dA data. ddRAD mtDNA on sp.based Cactoblastis oms 2.44-58.7924 -25.8424 -68.5389 -65.4058 -64.6124 -34.6639 -23.6014 -65.4970 -28.0242 Formosa -65.4334 Estero -25.0543 del Santiago -65.4058 (NWA) -23.6590 -67.1410 Jujuy -65.0382 -23.6014 -65.5706 Mendoza -36.4074 (NWA) -24.4775 -66.9374 Salta -36.2204 (NWA) Jujuy -36.2329 (NWA) -64.9460 Jujuy (NWA) -64.9460 Jujuy -25.2077 Pampa -65.4970 La -25.2077 Pampa La -65.2713 -25.0543 Pampa La (NWA) -24.3938 Salta (NWA) Salta (NWA) Salta (NWA) Jujuy pce nAgnia W:nrhetr retn.Nmesin Numbers Argentina. northwestern NWA: Argentina. in species 15 100608871 .2 .4 1.030 1.021 1.031 1.048 1.106 0.040 1.113 0.028 0.034 0.037 0.086 0.028 0.095 0.026 0.028 0.044 0.102 0.108 10 9 5 14 16 13 7 4 5 5 6 10 0.818 0.756 1 0.806 0.844 0.914 0.006 0.001 0.006 0.008 0.004 0.007 11 13 6 9 10 15 n 1f tteNB ne ceso number accession under NCBI the at v1.fa π .anacantha O. baumanii C. atacamensis T. sulphurea O. ficus-indica O. sulphurea O. sulphurea O. ficus-indica O. megapotamica O. megapotamica O. megapotamica O. quimilo O. ficus-indica O. ficus-indica O. ficus-indica O. (1) (10) (5) (6) (5) (2) (5) (1) (4) (4) (5) (10) (4) (6) (5) H d H n H e H o Ar Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. d 6502,3 31,894 5,159 36,657 27,047 0.00253 0.00678 0.00007 89,113 25,735 53,295 3,213 79,545 27,504 7,226,922 24,173 10,959,862 149,770 1,719 18,679 458,031 2,607,920 0.00098 0.00097 0.00001 53,807 26,550 23,399 3,712 40,952 32,329 563,085 3,501,657 25,829 133,561 1,340 2,461,656 176,482 9,297 0.00108 0.00356 0.00004 83,283 38,460 1,400,678 4,452,304 49,125 - 140,741 - - 2,418,707 - 311,765 1,441 12,999 0.00554 0.00286 0.00383 89,113 shown 10,959,862 each are 7,226,922 years with of associated millions 53,295 plant in host estimated method main times coalescent of divergence 79,545 using representation nodes; in esquematic reconstructed displayed an tree are and probability species SNPs, Posterior (c) 297,302 1,688 clade. and panels on inference, c based Bayesian and SNAPP by b, in inferred a, ddRAD - 332,320 in (b) codes and Color 461,118 - species. steps, 3. each 27,878 mutational Figure 101,474 of in green: locations, NWA number - Light geographic from the haplogroups same: are haplotypes respective major the haplogroups the three their - are within the denote shows in and squares network between individuals remarked The Values (c) and all species. panel. each of zoomed to a proportions as corresponding NWA admixture region sympatric of MDZ the charts to including Estero); 0.00003 assigned pie del individuals (Santiago shows indicates SDE Asterisk (b) 0.00003 Argentina; Pampa). (northwestern) Codes NWA (La LPP cluster/species. populations: corresponding (Mendoza); 0.00002 of the to origin individual geographic each of denote probability membership ( with 3,501,657 of clusters along individuals (2014) outside three shown Morrone the 53,807 563,085 of the are representation plant to according host 23,399 provinces new 2. Figure biogeographic a the on 40,952 represent feeding regions population Colored of moth regionalisation. the species area. 95,244 and three zoomed populations, the the allopatric where (NWA) the 128,466 Argentina of northwestern 108,731 to corresponds 1. 44,553 intervals. 3,742 Figure - confidence rate the migration outside M: fall legends - admixture; * Figure of by time indicated estimates Tadm: - Point generation; allopatric. per A: divergence - B: of time generation. 0.00496 per Tdiv: size; population 0.00153 Ne: 0.00040 4,452,304 4 adm 1,400,678 T 3 adm 83,283 T 2 adm 38,460 T 1 adm 49,125 T divergence intraspecific D-B 178,012 After M 5. C-B Model 398676* M C-D divergence 442,965 M intraspecific After 5 5. Tdiv 119082* Model 4 27,398 Tdiv divergence intraspecific 3 After Tdiv 5. Model 2 Tdiv divergence 1 intraspecific Tdiv Before 1. CS Model Ne CA divergence Ne intraspecific Before DS 1. Ne Model DA Ne divergence intraspecific BS Before Ne 1. Model parameters Estimated Fastsimcoal2. using simulations 3. Table H heterozygosity; size; Sample n: hlgntchptee n pce reifrne hlgntchptee ae n()mtDNA, (a) on based hypotheses Phylogenetic inference. tree species and hypotheses Phylogenetic on siae n 5 ofiec nevl fdmgahcprmtr nerdfo coalescent from inferred parameters demographic of intervals confidence 95% and estimates Point lseigaaye ae ndRD n egbrjiigntokbsdo tN.()Visual (a) mtDNA. on based network joining neighbor and ddRAD, on based analyses Clustering egahclctoswhere locations Geographic π: o bevdhtrzgst;A:Allcrcns;NA otwsenArgentina. Northwestern NWA: richness; Allelic Ar: heterozygosity; observed : uloiedvriy H diversity; Nucleotide atbatsbucyrus Cactoblastis .bucyrus C. on siaeLwrbudUprbudPitetmt oe on pe bound Upper bound Lower estimate Point bound Upper bound Lower estimate Point =3 k= akblue: Dark ; Cactoblastis D: ; d ersnigeach representing ) altp iest;H ubro altps H haplotypes; of number H: diversity; Haplotype : atbatsdoddi Cactoblastis .doddi C. p.mt ouain eesmld h omdarea zoomed The sampled. were populations moth spp. 16 n purple: and , Cactoblastis C: ; atbatscactorum Cactoblastis .doddi C. .cactorum C. Cactoblastis pce.Vria asrepresent bars Vertical species. edn on feeding . oxs.Locations coexist. .ficus-indica. O. :sympatric; S: ; e Expected : Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. Kr n ilo fyas(y) aesaecntutdb h aeo h pce,adtegeographic the and species, the of Pampa). name (La the LPP by (Mendoza); years lineages MDZ constructed thousand Estero); between are del in flow (Santiago Labels given gene SDE gene are which Argentina; (Myr). northwestern of time during NWA years divergence time amount distribution. of and of relative million Admixture period the and the occurred. denote (Kyr) delimit populations to lines sympatric intended discontinuous between Red is flow incongruence. arrows divergence. mito-nuclear dotted intraspecific in before/after of involved were Size (b) simcoal2. and 2). (a) Figure 4. in in Figure assigned remarked colors Clades to correspond parenthesis. codes in (Color (a) panel in iulrpeetto fdmgahchsoyuigcaecn iuain nerdb Fast- by inferred simulations coalescent using history demographic of representation Visual

Latitude -40 -30 -20 -70 -60

Longitude Moth species 17 Moths on new host plant Moths onnewhost Cactoblastis doddi Cactoblastis cactorum Cactoblastis bucyrus Cactoblastis -66.5 -65.5 -64.5 -50 Biogeographic provinces Yungas Puna Pampean Chacoan Monte -23.5 -24.5 -25.5 Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (a) (b) (a)

C. doddi C. cactorum C. bucyrus Admixture coefficients Admixture * * * * 1 0.8 0.6 0.4 0.2 0 MDZ NWA LPP NWA SDE NWA

Latitude -38 -31 -24 -71 -67 -63 Longitude MDZ NWA 18 LPP SDE -59 -55 -51 (c) 9 Salta 34 7 40 5 Jujuy Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (a) 3.41 1 0.99 2.33 1.79 1 1 0.63 1 0.41 0.28 C. doddi (NWA) C. bucyrus(NWA) Tucumania sp. C. cactorum(NWA) C. bucyrus(SDE) C. doddi (MDZ) C. cactorum (LPP) 19 (b) 1 1 1 1 C. doddi (NWA) C. doddi (MDZ) C. cactorum (LPP) C. bucyrus(NWA) C. cactorum(NWA) C. bucyrus(SDE) Posted on Authorea 15 Mar 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161578257.72978696/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. 2.2 Myr 2.2 ecological speciation Gene flowduring 0.7 Myr 0.7 Interspecific divergence Pleistocene 41.6 Kyr 41.6 Reproductive isolation 20 24.5 Kyr 24.5 19.2 Kyr 19.2 Intraspecific divergence 16.1 Kyr 1.8 Kyr 1.8 Kyr 16.1 secondary contact Gene flowin Holocene Rep. isol. (MDZ) C. doddi (SDE) C. bucyrus (NWA) C. doddi (LPP) C. cactorum (NWA) C. cactorum (NWA) C. bucyrus