A phylogenetic investigation of the Neotropical genus Alphamenes van der Vecht, 1977 (Hymenoptera, Vespidae, Eumeninae)

REVISTA

BRASILEIRA

DE

Entomologia

w w w . r b e n t o m o l o g i a . c o m

Systematics,

Morphology

and

Biogeography

A

phylogenetic

investigation

of

the

Neotropical

genus

Alphamenes

van

der

Vecht,

1977

(Hymenoptera,

Vespidae,

Eumeninae)

Letícia

A.

de

Oliveira

_,

_Tiago

_G.

_Inez

_,

_Wellington

_D.

_Ferreira

_,

_Marcel

_G.

_Hermes

a_{UniversidadeFederaldeLavras,DepartamentodeBiologia,LaboratóriodeSistemáticaeBiologiadeInsetos,Lavras,MG,Brazil} b_{UniversidadeFederaldaBahia,Salvador,LaboratóriodeEnsino,FilosofiaeHistóriadaBiologia,BA,Brazil}

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received24October2018 Accepted27November2018 Availableonline19December2018 AssociateEditor:JamesCarpenter Keywords: Malegenitalia Minixi Systematics Taxonomy

a

b

s

t

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a

c

t

Advancesinpotterwaspsystematicshavebeenachievedrecently,withclassificatorychangesresulting fromanalysesbaseduponlargescalemoleculardatasets.FortheNeotropics,recenthypothesespointto theoccurrenceofanexclusivecladerecognizedwithinthetribeEumenini.Inthisgroup,several contribu-tionsregardingtaxonomyandsystematicshavebeenproposedinthelastfiveyears,includingthegenus Alphamenes.ThistaxoncontainssevendescribedspecieswhosedistributionisexclusivelyNeotropical. Femalesaremorphologicallyhomogeneous,andcharactersrelatedtocopulatoryorgansareusefulinmale diagnosis.ThiscontributionformsthefirstphylogeneticapproachtoincludeallspeciesofAlphamenes, hencethefirsttostronglytestforgroupmonophyly.OurcladisticresultsrecoveredAlphamenesasa monophyleticgroupsupportedbymalegenitalfeatures.Relationshipsamongincludedspeciesalsorely upongenitaliccharacters,highlightingtheimportanceoftheseattributesforeumeninesystematics. RecentphylogeneticinvestigationsappliedtotheNeotropicalfaunaofpotterwaspsrepresentdesirable advancementstowardsanaturalclassificationforthegroup.

©2018PublishedbyElsevierEditoraLtda.onbehalfofSociedadeBrasileiradeEntomologia.Thisisan openaccessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/

).

Introduction

Phylogeneticstudiesbaseduponmoleculardatahaverecently changed our understanding of the evolutionary relationships withintheeumenines,theso-calledpotterwasps(Banketal.,2017;

Hinesetal.,2007;Piekarskietal.,2018).Thesestudiescontradict

ourtraditionalviewofthevespidwasp–andbyextent, relation-shipswithintheeumenines–phylogeneticrelationshipsbasedon morphologicaldata(Carpenter,1982;Hermesetal.,2014)anda totalevidenceapproach(PickettandCarpenter,2010),rendering Eumeninaeparaphyletic.

IntheNeotropicalregion,advancesinthesystematicsofthe groupwereachievedonly recently(e.g.HermesandCarpenter,

2012;HermesandOliveira,2016).Despitethefactthatthetribal

classificationproposedbyHermesetal.(2014)waschallengedby molecularstudies,theirresultsindicatethatasubgroupof Eumeni-naemayindeedbeanexclusiveNeotropicalclade.Thiscladewas recognizedas“Clade4”intheircontribution,whichcontainsthe majorityofthetaxonomicdiversitywithintheirtribeEumenini.

∗ Correspondingauthor.

E-mail:leticiabiologiaufla@gmail.com(L.A.Oliveira).

WithintheNeotropicalEumeninisensuHermesetal.(2014), some contributions encompassing a wide range of taxa were recently published (e.g. Grandinete et al., 2015; Hermes and

Oliveira, 2016). The eumenine classification within this region

hasbeencontinuallyinvestigatedundercladisticapproaches,with genericsynonymyresultinginthecasesofPachymenesde Saus-sure,1852andSantamenesGiordaniSoika,1990(Grandineteetal., 2015)andMinixiGiordaniSoika,1978andPachyminixiGiordani

Soika,1978(HermesandOliveira,2016).Regardingthelatter,a

specialcaseofacloselyrelatedtaxon,namelyAlphamenesvander Vecht,1977,remainstobecarefullyaddressed.Sexualassociation formembersofthisgenusisparticularlydifficult,withfemalesof somespeciesbeingmorphologicallysimilar(GiordaniSoika,1978;

Oliveira et al.,2017).Also, no phylogeneticstudy todate have

includedallspeciesofAlphamenes,thusitsmonophylyisstillopen tofurtherinvestigation.

Thepresent contributionenhancesourknowledgeaboutthe Neotropicalfaunaofeumeninewasps,aimingattestingthe mono-phylyofAlphamenesandrecoveringthephylogeneticrelationships among itsspecies. Thisforms ourfinal contributiontoa series ofpapersdealingwiththesystematicsofAlphamenesandMinixi

(HermesandOliveira,2016;Oliveiraetal.,2017),whosegeneric

statushave beentraditionallyquestionedbyeumenineworkers

https://doi.org/10.1016/j.rbe.2018.11.006

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

Table1

MorphologicalcharacterlistforthecladisticanalysesofAlphamenes.FiguresthatdonotappearinthetextareavailableinSupplementaryMaterial.

Head

(1)Clypealteeth:(0)carinate(Figs.16a,19and21);(1)ecarinate(Fig.17).

(2)Clypealapex,middleportion:(0)stronglyemarginate(Figs.16b,18b);(1)weaklyemarginate(Fig.17b). (3)Clypealdimensions:(0)widerthanlong(Fig.18c);(1)longerthanwide(Fig.16c).

(4)Labrumapex:(0)truncated(Fig.19e);(1)rounded(Fig.20e).

(5)Interantennalregion:(0)raisedandrounded(HermesandOliveira,2016,p.190,Fig.6);(1)raisedandlongitudinallycariniform(HermesandOliveira,2016,p.190,Fig. 7).

(6)Fronsmedially:(0)withfurrow(Figs.16d,17dand4d);(1)withoutfurrow(Fig.21d). (7)Maleantennae:(0)withtyloids(Figs.22a,23aand27a);(1)withouttyloids.

(8)Shapeofmaletyloids(applicableonlytoterminalsthatreceivedstate0forcharacter7):(0)narrow(Figs.22aand23a);(1)wide(Fig.27a). (9)F11ofmaleantennae:(0)long,reachingoralmostreachingtheapexofF9(Fig.22b);(1)short,clearlynotreachingtheapexofF9(Fig.25b). (10)ShapeofmaleF11:(0)pointed(Fig.24c);(1)rounded(Fig.26c).

Mesosoma

(11)Lateralsurfaceofpronotum:(0)depressed(HermesandOliveira,2016,p.191,Fig.13);(1)flattoconvex(HermesandOliveira,2016,p.191,Fig.14). (12)Pronotalcarinaofpronotoum:(0)complete(Fig.30a);(1)incomplete(Fig.31a).

(13)Pronotalcarina,humeralregion:(0)slightlysinuous(Fig.32b);(1)stronglysinuous(Fig.33b). (14)Malepronotum:(0)withoutfovea(Fig.34a);(1)withfovea(Fig.30c).

(15)MesepisternumI:(0)withoutepicnemialcarina;(1)withepicnemialcarina(Fig.32).

(16)MesepisternumII:(0)withtransversesculptureadjacenttomeso-metapleuralcarina(Figs.35and36b);(1)withouttransversesculptureadjacentto meso-metapleuralcarina(Fig.34b).

(17)MesepisternumIII:(0)stronglypunctate(Figs.35cand36c);(1)weaklypunctateorunpunctate(Fig.34c). (18)Longitudinalmesepisternalsulcus:(0)withoutcrenulae(Fig.29d);(1)withcrenulae(Fig.28d).

(19)Tegulae,posteriormargin:(0)slightlyconvex,posteriorlobewelldefined(Fig.37d);(1)stronglyconvex,rounded,posteriorlobeill-defined(Fig.38d). (20)Dorsalsurfaceoftegulae:(0)reticulate(Figs.37eand40);(1)slightlypunctate(Fig.38e);(2)stronglypunctate(Fig.39e).

(21)Sulcusbetweenmesoescutumandmetanotum:(0)highlydeveloped(Figs.42aand33a);(1)littleornotdeveloped(Fig.41a).

(22)Metanotum:(0)withlaterallongitudinalcarinaadjacenttodisc(Figs.42band43b,44b);(1)withoutlaterallongitudinalcarinaadjacenttodisc(Fig.45b). (23)Puncturesonlateralregionofpropodeum:(0)evident;dense(Fig.46a);(1)obsoleteorabsent(Fig.34f).

(24)Propodeum:(0)swollendorsolaterally(Figs.50aand53);(1)notswollendorsolaterally(Figs.51aand52a).

(25)Mediansulcusonposteriorsurfaceofpropodeum,lowerportion:(0)deeplydepressed(Fig.49b);(1)notdeeplydepressed(Fig.47b).

(26)Medianconcavityonposteriorsurfaceofpropodeum:(0)long,reachinghalfofpropodeum(Fig.48c);(1)short,notreachinghalfofpropodeum(Fig.49c). Metasoma

(27)T1indorsalview:(0)graduallyexpandingtowardsapex(Figs.51b,52band54c);(1)abruptlyexpandingmedially(Fig.57c);(2)abruptlyexpandingtowardsapex, campanulate(Figs.50band53c);(3)expandedallalong(Fig.56c).

(28)T1:(0)stronglypunctate(Figs.50c,52c,53a,54aand55a);(1)slightlypunctate(Figs.51cand57c). (29)ApicaldepressiononT1:(0)stronglydeveloped(Fig.57b);(1)slightlydevelopedorabsent(Figs.53band56b). (30)T2shapeindorsalview:(0)rounded,aswideaslong(Fig.58a);(1)elongate(Fig.59a).

(31)PunctationonT2:(0)denseanddeep(Fig.58b);(1)scarceandshallow(Fig.59b). (32)ApexofT2:(0)withlamela(Fig.60c);(1)withoutlamela(Fig.61c).

(33)FemaleS2:(0)denselysetose(Fig.62d);(1)sparselysetose(Fig.63d).

(34)MaleS7:(0)withoutcarina(Fig.4and65);(1)withcarina(Figs.2,3,5,6,7and64)

(35)MaleS7,shapeofcarina(applicableonlytoterminalsthatreceivedstate1forcharacter34):(0)withlongitudinalmediancarina(Figs.2and3);(1)withtransverse mediancarina(Figs.6and7).

Malegenitalia

(36)Ventrallobiofthepenisvalves;shape:(0)single,onethintooth;(HermesandOliveira,2016,p.193,Fig.32);(1)bifid,twoteeth(Figs.10and11);(2)morethantwo teeth,marginserrate(Figs.12and13);(3)single,oneenlargedtooth(Fig.15);(4)single,onetoothsomewhatenlarged(GiordaniSoika,1978,p.406,Fig.485);(5) slightlyevident,veryshort(Hermes;Carpenter,2012,p.15,Fig.37).

(37)Apexoftheedeagus:(0)withsulcus(Fig.67(HermesandOliveira,2016));(1)withoutsulcus(Fig.66(HermesandOliveira,2016)).

(38)VentralmarginofaedeagusI:(0)smoothatmedialenlargement(HermesandOliveira,2016,p.193,Fig.32);(1)serrateatmedialenlargement(Fig.9d) (39)VentralmarginofaedeagusII:(0)withacessorylobe(Fig.9e);(1)withoutacessorylobe(HermesandOliveira,p.193,Fig.32).

(40)Basalplateofaedeagus:(0)shorter(sometimesveryshort)thanbasalapodemes(Figs.9–15);(1)elongate,longerorreachingbaseofbasalapodemes(Hermesand Oliveira,2016,p.193,Fig.32).

(41)Digitus:(0)withoutmedianprojectionadjacenttolamella(HermesandOliveira,2016,p.193,Fig.34);(1)withmedianprojectionadjacenttolamella(Fig.8a). (42)Cuspis:(0)withoutmedianstrongsetae(HermesandOliveira,2016,p.193,Fig.34);(1)withmedianstrongsetae(Fig.8b).

inthelastcoupleofdecades(seeCarpenterandGarcete-Barrett,

2003).

Materialandmethods

Materialexaminationandmorphologicalstudies

We examined 376 specimens belonging to Alphamenes and Minixi(Table3inSupplementaryMaterial).Thespecimenswere borrowed from the following institutions: AMNH – American Museum of Natural History, New York, USA (Dr. James M. Carpenter);MCZ–MuseumofComparativeZoology,Harvard Uni-versity,Cambridge, USA(Dr.JignashaRana);MNHN – Muséum Nationald’HistoireNaturelle,Paris,France(Dr.ClaireVillemant); MNHNPY – Museo Nacional de Historia Natural del Paraguay, San Lorenzo, Paraguay (Dr. Bolívar R. Garcete-Barrett); CMNH

– Carnegie Museum of Natural History, Pittsburgh, USA (Dr. John Rawlins); NHM – Natural History Museum, London, Eng-land (Dr. Gavin Broad); ZMHB – Museum für Naturkunde der Humboldt-Universität,Berlin,Germany(Dr.FrankKoch);MSNVE –Museo di StoriaNaturaledi Venezia,Venice, Italy (Dr.Marco Uliana); HYMSJRP – Colec¸ão de Hymenoptera, Departamento de Zoologia eBotânica, Instituto de Biociências, Letras e Ciên-cias Exatas, Universidade Estadual Paulista ‘Júlio de Mesquita Filho’, São José do Rio Preto, Brazil (Dr. Fernando B. Noll); UFMG–Colec¸õesTaxonômicasdaUniversidadeFederaldeMinas Gerais, Belo Horizonte, Brazil (Dr. Fernando A. Silveira); DZUP – Colec¸ão de Entomologia Pe. Jesus Santiago Moure, Departa-mentodeZoologiadaUniversidadeFederaldoParaná,Curitiba, Brazil (Dr. Gabriel Melo); CEUFLA – Colec¸ão Entomológica da UniversidadeFederaldeLavras,Lavras,Brazil(Dr.MarcelG. Her-mes).

Table2

Charactermatrixusedforphylogeneticanalyzes.Thesymbols‘–’and‘?’correspondtoinapplicableandunobserveddata,respectively.

Characters Terminals 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 3 4 4 4 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 Monobiaangulosa 1 1 1 1 1 0 1 – 0 1 1 0 0 1 1 1 0 1 0 0 0 0 0 1 1 1 3 1 1 0 1 1 1 1 – 5 0 0 1 1 0 0 Minixibrasilianum 0 0 1 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 1 0 1 1 0 1 1 1 0 0 1 0 0 0 0 1 – 0 0 0 1 1 0 0 Miniximexicanum 1 0 1 0 0 0 0 1 0 0 0 0 1 1 1 1 0 1 1 0 1 1 0 1 1 1 0 1 1 0 1 0 0 1 1 ? ? 0 1 1 0 0 Minixisuffusum 1 0 1 0 0 0 0 1 0 0 0 0 1 1 1 1 0 1 1 0 1 1 0 1 1 1 0 0 1 0 0 0 0 1 – 0 0 0 1 1 0 0 Minixitricoloratum 0 0 1 0 1 0 0 1 1 0 0 0 1 1 1 1 0 1 1 2 1 1 0 1 1 1 0 0 1 1 1 0 0 0 – 0 1 0 1 1 0 0 Minixiarechavaletae 0 0 1 0 1 0 0 1 0 0 0 0 0 1 0 0 0 1 1 0 1 1 0 0 0 1 2 0 1 1 1 1 1 0 0 0 1 0 1 1 0 0 Minixisumichrasti 0 0 1 0 1 0 0 0 0 0 0 0 0 1 0 1 0 1 1 0 1 1 0 0 0 1 2 0 1 1 1 1 1 1 – 0 1 0 1 1 0 0 Minixiuruguyense 0 0 0 0 1 0 ? ? ? ? 0 0 0 1 0 0 0 1 1 0 1 1 0 0 0 0 2 0 1 1 1 0 1 ? – ? ? ? ? ? ? ? Minixibifasciatum 1 0 1 0 1 0 0 1 0 1 0 0 0 1 0 0 0 1 1 0 1 1 0 0 0 1 2 0 1 1 1 1 1 1 – 4 1 0 1 1 0 0 Minixijorgenseni 0 0 1 0 1 0 0 1 0 0 0 0 0 1 0 1 0 1 1 1 1 1 0 0 0 1 2 0 1 1 1 1 1 1 – 0 1 0 1 1 0 0 Minixibrethesi 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 1 0 1 1 0 0 1 0 0 0 0 2 0 1 1 1 1 0 1 – ? ? ? ? ? ? ? Minixiatrum 0 0 1 0 ? ? 0 0 0 0 ? 0 ? ? ? ? ? ? 1 0 ? ? 0 0 1 ? 0 0 1 1 1 0 ? ? ? ? ? ? ? ? ? ? Miniximariachii 0 0 1 0 0 1 0 1 0 0 0 0 0 1 0 1 0 1 1 0 1 1 0 0 1 1 0 1 1 1 1 0 1 1 – 0 0 0 1 1 0 0 Alphamenescampanulatus 0 0 1 0 1 0 0 0 0 0 1 0 0 1 1 1 0 0 1 0 1 0 0 1 1 1 0 1 1 0 1 1 0 0 1 0 0 1 0 0 1 1 Alphamenesincertus 0 0 1 0 1 0 0 0 0 0 1 0 0 1 1 1 0 0 1 0 ? 0 0 1 1 1 0 1 1 0 1 1 ? 0 1 2 0 1 0 0 1 1 Alphamenesinsignis 0 0 1 0 0 0 0 0 0 0 1 0 0 1 1 1 0 0 1 0 1 0 0 1 1 1 0 0 1 1 1 1 1 1 – 3 0 1 0 0 1 1 Alphamenesrichardsi 0 0 1 0 0 1 0 0 0 0 1 0 0 1 1 1 0 0 1 1 0 0 0 1 1 1 0 0 1 1 1 1 1 1 – 0 0 1 0 0 1 0 Alphamenesusitatus 0 1 1 0 0 0 0 0 0 0 1 0 0 1 1 1 0 0 1 0 1 0 0 1 1 1 0 1 1 1 1 1 1 0 1 2 0 1 0 0 1 1 Alphamenesconvexus 0 ? ? ? ? ? 0 0 0 0 1 0 ? 1 1 ? 0 ? 1 ? ? ? 0 1 1 ? 0 1 1 0 1 ? ? 0 0 1 0 0 0 0 1 1 Alphamenessemiplanus 0 0 1 ? ? ? 0 0 0 0 1 0 ? 1 1 ? 0 ? 1 0 ? ? 0 1 1 ? 0 1 1 1 1 ? ? 0 0 1 0 0 0 0 1 1 Pachymenesater 0 0 1 1 1 1 0 0 0 0 1 1 – 0 0 1 1 0 1 0 1 1 1 1 1 1 1 1 0 1 1 1 1 0 0 0 1 0 1 1 0 0 Pachymenessericeus 0 0 1 1 1 0 ? ? ? ? 1 1 – 0 0 1 1 0 1 0 1 1 1 1 1 1 1 1 0 1 1 1 0 ? ? 0 1 0 1 1 0 0 Pachymenespicturatus 1 0 1 0 0 1 0 1 0 0 1 1 – 0 0 1 1 0 0 0 1 1 1 1 1 1 0 1 1 1 1 1 0 0 0 0 1 0 1 1 0 1 Laevimeneslaevigatus 0 0 1 0 1 0 0 0 0 0 1 0 0 1 1 1 1 0 1 0 1 1 1 1 1 1 0 1 1 1 1 1 1 1 0 1 1 0 0 1 0 1 Laevimenesmorbilosus 0 0 1 0 1 0 0 0 0 0 1 0 0 1 1 1 1 0 1 0 1 1 1 1 1 1 0 1 1 1 1 1 1 1 0 1 1 0 0 1 0 1

Themorphologicalstudyofthespecimenswascarriedoutwith thestereoscopicmicroscopeLeicaS8APO.Imagesofthestructures ofphylogeneticandtaxonomicinterestwereobtainedwithaLeica 10445929digitalcâmeracoupledtotheLeicaDFC295stereoscope microscope.TerminologyformalegenitaliafollowedBitsch(2012)

andforexternalmorphologyCarpenterandGarcete-Barrett(2003). Themalegenitaliawasdetachedfromthemetasomaandclarified on10%NaOHsolutionfor24–36h,thenneutralizedwithacetic acid,washedinwaterandstoredinglycerol.

Cladisticanalyses

For the reconstruction of thephylogenetic relationships, all recognizedspecies withinAlphamenes andMinixiwereincluded asingroup(sensuHermesandOliveira,2016).In thecase ofM. atrum Selis, 2017, the information was extracted from the lit-erature(Selis,2017).We basedouranalysesupon ourprevious matrix(HermesandOliveira,2016),withtheinclusionofallspecies of Alphamenes and adjusting charactercoding accordingly. This resultedinamatrixwith42morphologicalcharacters,anincrease ofeightcharactersfromHermesandOliveira(2016).Speciesofthe closelyrelatedLaevimenesGiordaniSoika,1978andPachymenes deSaussure,1852wereincludedintheanalyzes. Theinclusion ofthesetaxaintheanalyzesaimedattestingthemonophylyof theingroupgenera(mainlyAlphamenesandMinixi).Despitethe factthatMinixiwasalreadyinvestigatedphylogenetically(Hermes

andOliveira,2016),weoptedforincludingitsspeciestoperforma

strongertest(moreingrouptaxaandmorecharacters)ofthe mono-phylyofthetaxaunderscrutiny.Fortherootingofthetreesthe outgroupmethodwasused(NixonandCarpenter,1993)andthe speciesMonobiaangulosadeSaussure,1852wasselected.

Fitch’sparsimony(Fitch,1971)wastheoptimizationcriterion used in the present study. Character matrix construction was carriedoutwithWincladav.1.00.08(Nixon,1999–2002).Heuristic searchesforthemostparsimoniouscladogramswereperformed

with TNT v. 1.5 (Goloboff and Catalano, 2016), using implied

character weighing (Goloboff, 1993; Goloboff et al., 2008).The valuefor theconstantk wascalculated withthescriptsetk.run (k=3.046875).InTNTthe“NewTechnologySearch”option was used following Hermes et al. (2014) and Hermes and Oliveira

(2016).

The visualization and editing of the cladograms, as well as optimizationofthecharacters,wereperformedinWinclada,with onlyunambiguouschangesshown.Supportforthebrancheswas investigated through Symmetric Re-sampling (Goloboff et al., 2003)usingTNT1.5,with1000replicationsfortraditional(TBR) treesearchand10,000re-samplingreplications.

Resultsanddiscussion

The morphological character list is shown in Table 1. The phylogenetic analysis with implied character weighing of the matrix (Table 2) containing 42 characters returned one most-parsimonioustree(Fig.1),withlength(L)=100,consistencyindex (CI)=49, andretentionindex (RI)=72.However,theresampling analysispresentedsmallchangeswhencomparedtotheimplied weightingtree.ThesechangesarerelatedtotheplacementofM. tricoloratum(Zavattari,1911),M.mexicanum(deSaussure,1857) andM.brethesi(Bertoni,1927)(Fig.1).

Thegenericrelationshipsrecoveredhereinareidenticaltothe hypothesispresentedbyHermesetal.(2014).However,onlyone synapomorphy(vs.fivesynapomorphiesrecoveredbyHermesand

Oliveira(2016))andtwohomoplasies(vs.onerecoveredbyHermes

andOliveira(2016))supportMinixiasanaturalgroup.Ofthese,the

interantennalregionraisedandrounded(char5[0])andthelateral surfaceofpronotumdepressed(char11[0])wererecoveredinboth studies.Itisworthyofmentioningthefactthattherelationships amongspeciesincludedinMinixichangedfromourpreviousresults

(seeHermesandOliveira,2016).Thehypothesisrecoveredherein

allowstherecognitionoftwomajorclades,whichwouldalsoallow theirtreatmentasseparategenera(forexample,returningtothe conceptofMinixiandPachyminixiofGiordaniSoika(1978),withthe

Fig.1. Mostparsimonioustreeobtainedwithimpliedweighingofthecharacters(k=3.046875),withlength(L)=100;consistencyindex(CI)=49;retentionindex(RI)=72. Blackrectanglescorrespondtouniquetransformations(synapomorphies)andwhiterectanglescorrespondtohomoplastictransformations.Supportvalues(symmetric resampling)aregivenbelowbranches(GCvalues)(wherevaluesaremissing,branchescollapsedduringtheresamplinganalysis).Onlyunambiguouschangesareshown.

inclusionofM.mariachiiandM.atruminthelatter).Nevertheless, weadvocatetheconceptofHermesandOliveira(2016)and recog-nizeonlyonegenus,basedonthelowresamplingsupportforthe M.atrum–M.bifasciatum(vonShculthess,1904)clade(Fig.1)and thetransitionalmorphologicalconditionofM.mariachiiHermes

andOliveira,2016(seeHermesandOliveira,2016fordetails).

Alphameneswasrecoveredasmonophyleticwithasomewhat highsupportvalue.Also,thisisthefirststudytoincludeall rec-ognizedspecieswithinthetaxoninaphylogeneticinvestigation. Twosynapomorphiesrelatedtomalegenitaliasupporttheclade:

basalplateofaedeagusshort(char40[0])anddigituswithmedian projectionadjacenttolamella(char41[1])(Figs.8–15).Themale genitaliahasalreadybeenpointedasprovidingstrongevidencefor themonophylyofAlphamenes(HermesandOliveira,2016;Oliveira etal.,2017).Furthermore,featuresassociatedwiththemalelast visiblesternum(Figs.2–7)aretaxonomicallyimportantand

reli-able(GiordaniSoika,1978).Ontheotherhand,identificationto

speciesleveloffemalesofAlphamenesisquitedifficult,andonly recentlyOliveiraetal.(2017)providedfurthermorphological evi-dencetodistinguishsomespecieswithinthegenus.

Figs.2–7.Malesternum7(arrows):2.Alphamenesconvexus(Fox,1899);3.A.semiplanusGiordaniSoika,1978;4.A.insignis(Fox,1899);5.A.campanulatus(Fabricius,1804); 6.A.incertus(deSaussure,1875);7.A.usitatus(Fox,1899).Figs.2,3,4,5and7modifiedfromOliveiraetal.,2017.ScalebarsforFigs.2,3,4,5=1mm;Figs.6,7=0.5mm.

WithintheAlphamenes,twomajorcladesmaybereadily recog-nized.Nevertheless,thesearebothsupportedonlybycopulatory features,suchastheshapeofthelastmetasomalsternum’scarina

(Figs. 2–7)and featuresrelated totheaedeagus(Figs.8–15).In

general,fewsynapomorphieswererecoveredtosupportinternal relationshipswithin Alphamenes, especially because most char-acters were extracted from females. As mentioned previously

(GiordaniSoika,1978;HermesandOliveira,2016;Oliveiraetal.,

2017),femalehomogeneityhamperstaxonomyofthegenus,butis alsoanotherevidenceformonophylyofthegroup.

Conclusion

Thecharactermatrix andthetaxonsamplingin thepresent studywereincreasedfrompreviousinvestigations,whichresulted inamorerobusttestforthemonophylyoftheincludedtaxa.This isthefirstcontributiontoincludeallspecies recognizedwithin Alphamenes,andthemonophylyofthetaxonwasrecoveredwith somewhatstrongsupport.Asmentionedinpreviousstudies, char-acters ofmale genitaliawere importantin recoveringboth the monophylyofAlphamenesaswellasinternalrelationshipsamong itsspecies.

ThetaxonomyofAlphamenesmayberegardedassatisfactorily resolved,despitesomefemalesarestilldifficulttodifferentiate(e.g.

A.semiplanusGiordaniSoika,1978,A.incertus(deSaussure,1875), A. campanulatus (Fabricius,1804) and A. convexus (Fox, 1899)). Someeffortsmayallowtheresolutionofthesematters,suchas fieldworkinordertorecovernestsofAlphamenesformalesand femalesmaybereared(Oliveiraetal.,2017).Fromthere,several morphologicalapproachesmaybeconductedtosexuallyassociate individuals of a given species, suchas internal anatomy of the exoskeleton,femalesting,andmouthparts.Finally,anintegrative approachforAlphamenestaxonomywouldmostcertainlyserveas asolidbasisforrobustspecieshypotheseswithinthegenus.

Aseriesofrecentcontributionswererecentlypublishedbythe authors(e.g.Ferreiraetal.,2015,2017,2018;HermesandOliveira,

2016;Oliveiraetal.,2017)regardingNeotropicaleumenine

tax-onomy andphylogeny. The continuingpracticeof investigating thegenericlimitsofNeotropicaltaxausingphylogenetictoolsisa desirableone,especiallyaftertheexpansionofgenericnames con-ductedbyGiordaniSoikabetweenthe1970sand1990s.Examples of obscure and/or dubious recognition of genera are Ancistro-ceroidesde Saussure,1855,Hypancistrocerus de Saussure,1855, HypodynerusdeSaussure,1855(especiallytheeastern Neotrop-icalfauna),ParancistrocerusBequaert,1925,andStenodynerusde Saussure,1863.Phylogeneticinvestigationsfollowedbymodern taxonomicrevisionsaredesiredforthesetaxa,forthesomewhat simpletaskofdescribingnewspeciesbeconductedwithclear con-ceptsaboutgenericlimitswithintheNeotropicaleumenines.

Figs.8–15.Malegenitalia.Figs.8and9.A.campanulatus(Fabricius,1804):8.Gonocoxiteandvolselainlateralview;9.Aedeagusinlateralview(a.digitus;b.cuspis;c. ventrallobiofthepenisvalves;d.ventralmarginofaedeagus;e.acessorylobe;f.basalapodemeofaedeagus;)(modifiedfromHermesandOliveira,2016).Figs.10–15. Ventrallobiofthepenisvalves(aedeagus):10.A.convexus(Fox,1899);11.A.semiplanusGiordaniSoika,1978;12.A.incertus(deSaussure,1875);13.A.usitatus(Fox,1899); 14.A.richardsiGiordaniSoika,1978;15.A.insignis(Fox,1899).ScalebarsforFigs.10–12and14–15=0.2mm;Fig.13=0.5mm.

Conflictsofinterest

Theauthorsdeclarenoconflictsofinterest. Acknowledgements

WethankthecuratorscitedintheMaterialandMethods sec-tionfortheloaningofthespecimens.ThankstoDr.SalvadorArias forprovidingthesetk.runscriptandtheWilliHennigSocietyfor allowingtheuseofthesoftwareTNT.LAOandWDFaresupported byFAPEMIGandCNPqscholarships,respectively.Financialsupport wasprovidedbyFAPEMIGProcessCRA-APQ-02784-17.

AppendixA. Supplementarydata

Supplementarydataassociatedwiththisarticlecanbefound,in theonlineversion,atdoi:10.1016/j.rbe.2018.11.006.

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