EUEURROOPEAPEANNJJOOUURRNNAALLOOFFEENNTTOOMMOOLLOOGYGY ISSN(online):1802-8829

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EU EU R R O O PEA PEA N N J J O O U U R R N N A A L L O O F F E E N N T T O O M M O O L L O O GY GY

ISSN(online):1802-8829

http://www.eje.cz Eur.J.Entomol.115:387–

399,2018doi:10.14411/ej e.2018.039

ORIGINALARTICLE

DiscoveryofaremarkablenewspeciesofLymanopoda(Lepidoptera:

Nymphalidae:Satyrinae)andconsiderationsofitsphylogeneticpositio n:Anintegrativetaxonomicapproach

TOMASZW.PYRCZ ^1, ⁵,CARLOSP R I E TO ^2, ³,PIERREB O Y E R ⁴and

JADWIGAL O R E N C - B R U D E C K A^5, * 1EntomologyDepartment,InstituteofZoologyandBiomedicalResearch,JagiellonianUniversity,ul.Gronostajowa9,30- 387Kraków,Poland;e-mail:tomasz.pyrcz@uj.edu.pl

2CorporaciónUniversitariaAutónomadelCauca,Calle5#3-87,Popayán,Colombia;e-mail:cprieto50@gmail.com 3SNSB-BavarianStateCollectionofZoology,Münchhausenstr.21,81247Munich,Germany

47,Lotissementl’Horizon,13610LePuySainteRéparade,France;e-mail:pierdom@aliceadsl.fr 5NatureEducationCentre,JagiellonianUniversity,ul.Gronostajowa5,30-387Kraków,Poland;e- mail:j.lorenc-brudecka@uj.edu.pl

Keywords.Lepidoptera,Nymphalidae,Satyrinae,Lymanopoda,systematics,newspecies,phylogeny,DNAbarcodes,biodiver sity,páramo,Colombia,Andes,ValledelCauca

Abstract.AnewspeciesofLymanopodaWestwood,acloudforestNeotropicalgenusofSatyrinae,isdescribedfromthepáramograsslandson anisolated,periphe ral massif intheColombian Central Cordiller a oftheAndes:L. flammigeraPyrcz, Prieto&Boyer,sp.

n.ThegenusLymanopodaisspecies-

rich(approx.65species)anditsalphataxonomyisrelativelywellresearched.Relationshipswithinthegenususingmoleculardataha vealsobeenexplored.Thenewspeciesisoutstandingforitsgoldenyel-

lowcolourinmales,notfoundinanyotherneotropicalSatyrinae.CladogramswereconstructedbasedonCOIsequencesof47specieso fLymanopoda(~

70%oftheknownspecies)including17fromColombia.Thenewspeciessegregatesinthe“tolima”clade,whichcomprisesfouro therhighaltitudeColombianspecies,aswellastwofromEcuado r.However,itisthecompara-

tiveanalysisofmalegenitalia,inparticularthesuperuncusandvalvae,whichidentifieditsclosestrelatives,thusconfirmingthatgenitalc haracterscanhelprefinemolecularphylogenies.InadditiontoidentifyingspeciesusingmitochondrialDNA(mtDNAbar-

codes),nucleotidesiteswithuniquefixedstatesusedtoidentifyninespeciesofLymanopodafromColombiaarealsopresented.

ZooBankArticleLSID:F820B047-2E29-4DEC-9C23-BB9A5B076528

INTRODUCTION

Colourpatternsofbutterflywingsareamongthemost outstandingexpressions ofevolution.Undercertainc on-

ditionstheyareextremelyplasticandevendramaticch angescanbecontrolledbysimplegeneticmechanismsan dquicklyrespondtoselectivepressure,forexampleinH eliconius(Kronforst&Papa,2015).Colourshavemanyadap tiveroles,commonlyinintraspecificsexualcommu- nication,w a r n i n g , m i m e t i c r e l a t i o n s o r,v e r y f r e q u e n t l y,crypsis.I n d i fferentg r o u p s o f b u t t e r fliesd i ff erentr o l e s dominate.I n t h e c o s m o p o l i t a n s u b f a m i l y S a t y r i n a e , t h e prevailinga d a p t a t i o n i s c a m o uflage,a n d i t s o v e r 2 5 0 0 speciesareoverwhelmingly darkwithshadowsofbrown,thustheircommonEnglishname(

browns),withsomeele-

mentsenhancingtheircrypticcolouration,suchasstripesa ndpatchesimitatingthesubstrate, inmostcasesonth e

undersidesoftheirwings.Inafewgenera,suchasElym- niasHübner,1818andElymniopsisFruhstorfer,1907,show ycoloursare,however,dominant,whichisexplainedbytheirinvol vementinBatesianmimicryrings(Mallet&Joron,1999).A mongthefewSatyrinaegenerawithspe-

ciesbearingconspicuouswingpatchesistheneotropi calmontaneLymanopodaWestwood.Thisgenuscanbecon- sideredasoneofthebestknownamongSouthAmericanSat yrinaeandthereareanumberofpaperspublished,es- peciallyi n t h e l a s t t w o d e c a d e s , o n t h e i r t a x o n o m y a n d distribution(Pyrcz,1999,2003,2004,2005,2012;P yrcz

&B o y e r,2 0 11;P y r c z & R o d r í g u e z , 2 0 0 6 ; P y r c z e t a l . , 1999,2009a,b,2010,2016),phylogenetics(Casner&P yrcz,2010;Marínetal.,2016),ecology(Pyrcz&Woj- tusiak,2002;Pyrcz&Garlacz,2012)andbiology(Mon- tero&Ortíz,2012).Manyofthemorethan60speciesof

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*C o r r e s p o n d i n g author;e-mail:j.lorenc-brudecka@uj.edu.pl

Finalformattedarticle©InstituteofEntomology,BiologyCentre,CzechAcademyofSciences,ČeskéBudějovice.

AnOpenAccessarticledistributedundertheCreativeCommons(CC-BY)license(http://creativecommons.org/licenses/by/4.0/).

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Pyrczetal.,Eur.J.Entomol.115:387–399,2018 doi:10.14411/eje.2018.039 Lymanopodah a v e c o n s p i c u o u s w h i t e , b l u e , a n d r e

d d i s h groundcolours,whichmaybemarkedwithwhiteorgreen patches.T h e u n d e r l y i n g e v o l u t i o n a r y r a t i o n a l e f o r t h i s isstillunknownbutthereisgrowingev idencethatsomekindofmimicryisinvolved(Pyrcz,inprep.

).Yet,eveninLymanopodathediscoveryofaspecieswithshin ygoldenyellowmaleswasextremelysurprisingasthiskind ofco-

lourationisnotonlyuniqueforthegenusbutalsoamongall worldwideSatyrinae.Hereweinvestigateitsaffinitieswithi nthegenusLymanopodaandaddresssomequestionsaboutthea daptiveroleofitscolourpattern.

MATERIALANDMETHODS Morphologicalstudies

Mostoft he m a t e ri a l use d i n t hi s st ud y wa s o bt a i ne d duri ng field-

workb y C . P r i e t o a n d P.B o y e r i n C o l o m b i a . S p e c i m e n s us edf o r m o r p h o l o g i c a l s t u d i e s w e r e e x a m i n e d i n t h e N a t u r e EducationCentre(formerlyZoologicalMuseum) oftheJagiel-lonianUniversityinKraków(CEP-

MZUJ).Typesandadditionalspecimenswe re e x a m i n e d i n m a j o r pu bl i c m u s e u m s i n c l ud i n g InstitutodeCienciasNaturalesd elaUniversidadNacional,Bo-

gotá,C o l o m b i a ( I C N ) , t h e N a t u r a l H i s t o r y M u s e u m , L o n d o n , UK(NHMUK),MuseodeAgronomíadelaUniversidadCent ral,Maracay,Venezuela(MIZA),StaatlichesMuseumfürTierkunde,D resden,Germany(MTD)andZoologischeMuseum,HumboldtUnive rsität,Berlin,Germany(ZMHB),aswellasinthecollec- tionsofPierreBoyer(PB)andCarlosPrieto(RCCP).

Theterminalpartsoftheabdomens(includingthe genita lia)wereremovedfromthespecimensandsoakedin10%KOHso- lutionfor5–

10min.Subsequentl y,abdomenswerepreliminar- ilycleanedusingsofttissueinwaterinordertoexposegenital parts.Waterw a s r e m o v e d f r o m d i s s e c t e d g e n i t a l i a u s i n g 9 0 % and9 5 % s o l u t i o n s o f e t h a n o l . ANikond i g i t a l c a m e r a D S -

F i 1 anda nOl ym pus S Z X 9 st e re om i c rosc ope we re us e d f ort a ki ng picturesofthedissections,whichwerethenprocessedin AdobePhotoShop7. 0 C E a n d C o r e l P H O TO-

PAINTX3 p r o g r a m s t o enhancefocusandimprovequality.Thedi ssectedgenitaliawerekeptinglycerolinvialspinnedunderthec orrespondingspeci-

mens.GenitalterminologylargelyfollowsKlots(1956).Adult swerephotographedusingaMinoltaE-

500digitalcamera.Colourplatesw e r e c o m p o s e d u s i n g AdobeP h o t o S h o p v e r s i o n 8 . T h e followingabbreviationsareusedinthete xt:FW–forewing;HW

–h i n d w i n g ; D – d o r s u m ; V–v e n t e r ; H D P – h i n d w i n g d o r s a l medianpatch.

Materialandsamplingarea

PartialnucleotidesequencesofmtDNAcytochromecoxidasesubu nitIgene(COI)ofindividualsfromseveralpopulationsoc-

curringintheAndesinColombiathatwerepreviouslyidentifiedmorph ologically,wereanalyzed.Tissuesampleswereextractedfromi dentifiedpinnedspecimenscollectedinthepast10years,asitislessl ikelythatsequencedatacanbeobtainedfromoldma-

terial.Altogether79specimens,representing47species,yieldedaDN Asequenceofover400basepairs(bp)inlength.Specimenswithshorterseque nceswereexcludedfromtheanalyses.

Moleculardelimitationofspeciesandbarcodes

FortheDNAanalyses,79individualsof47speciesofLymano- podawereincludedaswellas2individualsoftwodifferentgen- eraasanoutgroup,CoradeschelonisHewitsonandLasiophi lazapatoza(Westwood).Oneortwolegswereremovedfromeachdrie dspecimenandstoredinindividualtubes.DNAextraction,ampli ficationandsequencingofthebarcoderegionoftheCOIgenew erecarriedoutattheCanadianCentreforDNABarcod-

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Pyrczetal.,Eur.J.Entomol.115:387–399,2018 doi:10.14411/eje.2018.039 ing(C C D B ), On t a ri o, C a n a d a , us i n gs t a nd a r d h i g h t hr o

ug hp ut protocols(Ivanovaetal.,2006;deWaardetal.,2008).PC Ram-

plificationwithasinglepairofprimersconsistentlyrecovereda65 8bpregionnearthe5’endofCOIthatincludedthestandard648b p b a r c o d e r e g i o n f o r t h e a n i m a l k i n g d o m ( H e b e r t e t a l . , 2004).Completespecimendataincludingimages,voucherdepo- sition,GenBankaccessionnumbers,GPScoordinates,sequencesandtra cefilesareaccessibleintheBarcodeofLifeDataSystem(BOLD) (Table3).

Sequencedivergencesforthebarcoderegionwerequantifie dusingtheKimura2Parametermodel,employingtheanaly ticaltoolsinBOLD(BOLDalignment,pairwisedeletion).T hiswasdonetodeterminewhetherthereisabarcodegap(abreakint hedistributionamonggeneticdistancesofspecimensbelongin gtothesamespeciesandthoseofspecimensfromdifferentspecies),t hatwouldallowtheidentificationofthespecimensexamine d.Geneticdistancesbetweenspeciesarereportedasminimumpair- wisedistances,whileintraspecificvariationisreportedasmean andmaximumpairwisedistances.

Severalquantitativespeciesdelimitationalgorithmsformolec- ulardatahavebeendevelopedoverthepastdecade,includingap-

proachesdedicatedtoDNAbarcodessuchasAutomaticBarcodeGapD i s c o v e r y ( A B G D ) a n d R e finedS i n g l e L i n k a g e ( R E S L ) Analysisalgorithm(Puillandreetal.,2012;Ratnasingham&He- bert,2013).Eachspecimenwithasequencelongerthan500 bpautomaticallygainsaBIN(BarcodeIndexNumber)assignme ntonBOLDthatisbasedontheRESLalgorithm(Ratnasingham&Hebe rt,2013).BINsmaybemergedwhengeneticallyintermedi-

atespecimensareadded,orsplitwhennewrecordsrevealaclearsequenc edivergencestructure.Distance-basedneighbour-

joining(NJ)wasusedtoreconstructDNA barcodegenetrees .Despitecertainlimitations,NJhasrepeatedlybeenshowntoperformwe llforspeciesidentification(Huelsenbeck&Hillis,1993;Kumar&Gad agkar,2000;Mihaescuetal.,2009;Mutanenetal.,2016).

Phylogeneticrelationships

Areconstructionofthephylogeneticrelationshipsof sp eciesofLymanopodawasdoneusingtheMaximunLikelihoo d(ML)method.TwospeciesofSatyrinaewereusedasanoutgroup:C o-

radeschelonisandLasiophilazapatoza.Theanalysiswasdo neusingthePhylogeny.frplatform(Dereeperetal.,2008,2010)andsequ enceswerealignedusingMUSCLE(v3.8.31)andconfiguredforhighestac curacy(MUSCLEwithdefaultsettings).

Thep h y l o g e n e t i c t r e e w a s c o n s t r u c t e d u s i n g t h e M L m e t h -

odi m p l e m e n t e d i n t h e P h y M L progra m( v 3 . 1 / 3 . 0 a L RT).T h e HKY85substitutionmodelwasselectedassuminga nestimatedproportionofinvariantsites(of0.601)andfourga mma-distrib-

utedratecategoriestoaccountforthepercentageheterogenei tyacrosssites.Thegammashapeparameterwasestimateddirectlyf romthedata(gamma=1.147).Thereliabilityofinternalbranch-

esw a s a s s e s s e d u s i n g t h e a L RTt est( S H -

L i k e ) . T he g r a p h i c a l representationa n d e d i t i n g o f t h e p h y l o g e n e t i c t r e e w e r e d o n e usingTreeDyn(v198.3).

RESULT S Taxono my

GenusLymanopodaWestwood,1851

LymanopodaWestwood,1851(May):Pl.LXVII,Figs6,7.Typespeci es:L.samiusWestwood,1851,bymonotypy.

SarromiaWestwood,1851(May):Pl.LXVII,Fig.5.Typespe- cies:S.obsoletaWestwood,1851,bymonotypy.SynonymizedbyWe stwood,1851(July):401–402.

ZabirniaHewitson,1877:92.Typespecies:Z.zigomalaHewit- son,1877,bymonotypy.SynonymizedbyPyrcz,2004:463.

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Fig.1.AdultsofLymanopodaflammigerasp.n.1.1–maleparatype(upperside);1.2–maleparatype(underside);1.3–

maleparatype(upperside);1.4–maleparatype(underside);1.5–femaleparatype(upperside);1.6–femaleparatype(underside).

TrophoninaRöber,1889:222.Typespecies:Lymanopodaacraei- daButler,1868,bymonotypy.SynonymizedbyPyrcz,2004:

463.

SabatogaStaudinger,1897:143.Typespecies:S.mirabilisSta udinger,1 8 9 7 , b y m o n o t y p y.S y n onymizedb y Adam s&

Bernard,1977:270.

LymanopodaflammigeraPyrcz,Prieto&Boyer,sp.n.

(Figs1.1–1.6,2.1–2.4,3.6)

ZooBanktaxonLSID:

4B29BAB2-E924-4C1B-AFCA-AF131920FF02 Diagnosis

Thisspecieshasthesize,wingshape,andwingpatternsi milartoL.huilanaWreymer,1911andL.tolimaWeymer,1890(dep ictedinFig.5),butmalesdifferfromboththesespeciesandfro m anyot hercongen erbyt hegol deny el -

lowcolouroftheirupperandundersides.Thefemalesare

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Fig.2.MalegenitaliaofLymanopodaflammigerasp.n.(paratype).

2.1–lateralview;2.2–viewfromabove;2.3–

aedeagusinlateralview;2.4–

detailsoftheapicesoftherightandleftvalvaeinlateralview.

whitishandthusnearlyinseparablefromthemostcloselyrela tedspecies,whichare,however,notsympatric.

Description Male.(Figs1.1–

1.4)Head:Eyeschestnutcoveredwithlong,b l a c k h a i r s ; l a b i a l p a l p s t w o a n d a h a l f t h e l e n g t h ofh e a d , c o v e r e d w i t h y e l l o w a n d b l a c k h a i r s , d o r s a l l y also brownscales;fronswithatuftofbrownhair;anten- naereachinghalflengthofthecosta,chestnutwithwhitesc alesatthebaseofeachflagellomere,clubcomposedof10s e g m e n t s , s t r o n g l y f latteneda n d d i l a t e d , b r o w n , d o r- sallyslightlylighterwithamediangroove.Thorax:Dor- sallyblack,mostlynaked,withsomelongbutsparsesilverhairs,tegu laecoveredwithlong,goldenbrownhairs;ven-

trallyblackbutcoveredwithlongand denseyellowan dwhitehairs;femoraofsecondandthirdpairoflegsblack,with firstpairandtibiaeandtarsiyellow,denselycoveredwithscales .Wings:FW(length:20–

21mm)triangularwithapointedapex,straightoutermarginan dshallowtornus;HWovalwitharounded apexandstra ightouter marginfromveinM2totornuswherebentnearlyat arightangle,analmarginstraight.FWDyellowofvariab leshade,be-

tweenpaleyellow(inolderindividuals)andgoldenyellowfrombasa ltopostmedianarea,exceptforagreyishbasalsuffusiona n d a n e l o n g a t e d p a t c h i n s u b a p i c a l a r e a ; d i s - tallydarkbrownwithsharpbasalnotchesalongthediscalcella ndveinCu2A,adarkbrownocellusinspaceCu1A-

Cu2A.HWDvaryingbetweenpaleyellowandgoldenyel-

loww i t h a g r e y i s h b a s a l a n d m e d i a l s u ffusiona n d w i t h

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Fig.3.Malegenitaliaof“tolima”clade(left:viewfromabove,right:lateral view).3.1–Lymanopodahuilanadominicae;3.2–L.hui- lanah u i l a n a ;3 . 3 – L . h u i l a n a s a l a z a r i ;3 . 4 – L . t o l i m a;3 . 5 – L . cassneri;3.6–L.flammigerasp.n.

aseriesofminute,submarginalblackdots(andinsome specimensmoreorlessdevelopedmarginaldarkpatche sbetweentornusandapex).FWVcolourpatternsimilart othato n t h e u p p e r s i d e , b u t t h e y e l l o w b a s a l a r e a i n v a r i -

ablylighter,andallthedarkbrownelementsaredullandb arelyvisibleexceptforthedarkerpatchinthepostdiscalarea.

HWV lighto ra ng ea l m os t l ac ki ng ap at t e rn ex c ep t fora l i g h t e r,e l o n g a t e d p a t c h i n d i s c a l c e l l a n d a d a r k e r brownareaimmediatelybehinddiscalcell;sub marginaltinyblackspotsasontheupperside.Abdomen:Blackd or-

sallyandlaterally(coveredwithdense,velvetblackhairsa ndscales),ventrallywithsandyyellowscalesandhairs.

Genitalia( F i g s 2 . 1 –

2 . 4 , 3 . 6 ) : Tegumens t r o n g l y s c l e r o -

tizedw i t h a s l i g h t l y b u l g e d d o r s a l s u r f a c e ; s

u p e r u n c u s prominent,reachinghalf lengthoftheun cus,bifurcated; uncusstoutwithasharptippointingdown wards;gnathosreduced,blunt;subscaphiumsmallandweaklyscler otized;

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Species Mean Intra-Sp

Max

Intra-Sp Nearestspecies Nearestneighbour

Coradeschelonis N/A 0 Lasiophilazapatoza GWOTU985-17

Lasiophilazapatoza N/A 0 Coradeschelonis GWOTU974-17

Lymanopodaacraeida N/A 0 Lymanopodavenosa GBGL8234-12

Lymanopodaaffineola N/A 0 Lymanopodaapulia GBMIN34627-13

Lymanopodaalbocincta 0.35 0.59 Lymanopodapanacea GWOTR709-16

Lymanopodaalbomaculata N/A 0 Lymanopodaapulia GBMIN34627-13

Lymanopodaaltis N/A 0 Lymanopodaconfusa GBMIN34624-13

Lymanopodaapulia N/A 0 Lymanopodaaffineola GBGL8210-12

Lymanopodaaraneola N/A 0 Lymanopodashefteli GBGL8232-12

Lymanopodacaeruleata N/A 0 Lymanopodacaucana GWOTR725-16

Lymanopodacaracara N/A 0 Lymanopodaflammigera GWOTR859-16

Lymanopodacasneri N/A 0 Lymanopodatolima GWOTR858-16

Lymanopodacaucana 0.14 0.21 Lymanopodacaeruleata GWOTU968-17

Lymanopodaconfusa N/A 0 Lymanopodaaltis GWOTR733-16

Lymanopodadietzi N/A 0 Lymanopodaaltis GWOTR733-16

Lymanopodaeubagioides N/A 0 Lymanopodainde GBMIN34617-13

Lymanopodaeuopis N/A 0 Lymanopodavenosa GBGL8234-12

Lymanopodaexcisa N/A 0 Lymanopodapieridina GWOTR758-16

Lymanopodaferruginosa N/A 0 Lymanopodashefteli GBGL8232-12

Lymanopodaflammigera 0 0 Lymanopodatolima GWOTR858-16

Lymanopodaflorenciensis 0.18 0.19 Lymanopodaaffineola GBGL8210-12

Lymanopodahazelana N/A 0 Lymanopodasamius GWOTR751-16

Lymanopodahuilana N/A 0 Lymanopodamelia GBGL8226-12

Lymanopodahyagnis N/A 0 Lymanopodaumbratilis GBGL8233-12

Lymanopodainde N/A 0 Lymanopodaeubagioides GBMIN34621-13

Lymanopodaionius N/A 0 Lymanopodapieridina GWOTR758-16

Lymanopodalabdassp. N/A 0 Lymanopodaaraneola GBGL8213-12

Lymanopodalecromi N/A 0 Lymanopodamaletera GWOTR722-16

Lymanopodamagna N/A 0 Lymanopodaobsoleta GWOTR712-16

Lymanopodamaletera 0 0 Lymanopodalecromi GBGL8224-12

Lymanopodamarianna N/A 0 Lymanopodalecromi GBGL8224-12

Lymanopodamelia N/A 0 Lymanopodahuilana GWOTR866-16

Lymanopodanadia N/A 0 Lymanopodaferruginosa GBGL8220-12

Lymanopodanevada 0 0 Lymanopodaparamera GWOTR874-16

Lymanopodanivea N/A 0 Lymanopodapieridina GWOTR758-16

Lymanopodaobsoleta 0.77 0.77 Lymanopodaconfusa GBMIN34624-13

Lymanopodapanacea N/A 0 Lymanopodaapulia GBMIN34627-13

Lymanopodaparamera 0 0 Lymanopodanevada GWOTR877-16

Lymanopodapieridina N/A 0 Lymanopodaionius GWOTR745-16

Lymanopodaprusia N/A 0 Lymanopodatolima GWOTR858-16

Lymanopodarana N/A 0 Lymanopodaumbratilis GBGL8233-12

Lymanopodasamius 0.1 0.15 Lymanopodahazelana GBMIN34619-13

Lymanopodashefteli N/A 0 Lymanopodahyagnis GBMIN34618-13

Lymanopodatolima 0 0 Lymanopodacasneri GWOTR869-16

Lymanopodaumbratilis N/A 0 Lymanopodahyagnis GBMIN34618-13

Lymanopodavenosa N/A 0 Lymanopodacaucana GWOTR726-16

Lymanopodavivienteni N/A 0 Lymanopodaferruginosa GBGL8220-12

Table1.Summaryofgeneticdistancesamong47speciesofLymanopoda.Foreachspecies,themeanandmaximumintra-specificval- uesarecomparedtothenearestneighbourdistance.Wherethespeciesisrepresentedbyasingleton,N/Aisdisplayedforintra-specificvalues.

Distance toNN 10.81 10.81 5.51 2.76 4.26 3.3 2.59 2.76 3.59 0.77 6.77 3.31 0.77 2.59 3.3 3.86 5.51 4.39 3.3 5.3 4.19 6.99 4.41 0.39 3.86 2.94 4.69 3.06 4.75 3.06 7.5 4.41 5.35 5.78 4.21 2.95 3.31 5.78 2.94 9.24 3.94 6.99 1.95 3.31 0.39 4.91 7.77

appendixangul a r,st out but short wi t h asharpt i p; val vaelongated,wideinbasalhalf,narrowerinthemiddle,endswith awideserratedapexandaprominentprocessuspoint-

ingu p w a r d s ; s a c c u s s h o r t a n d f l attenedd o r s o - v e n t r a l l y ; aedeagussimple,tubular,thevalva+saccusv eryslightlyarched,withasmoothsurface.

Female.(Figs1.5,1.6)Sexualdichroismprominent;yel-

lowisreplacedbywhitepigmentation,howeverthedark brown-

blackishelementsofthecolourpatternarenearlyidenti cal,exceptthattheyareslightlylargerontheFWD,enteringm oredeeplyintothediscalcell.TheHWDsubmarginal bl ac kd ot sa re a l s ol a rger.Ot he rw i s e, t h ew i n g shape ofthefemalediffersslightlyinbeinglesselongated,

especiallyt h e h i n d w i n g s ( F W l e n g t h : 2 1 m m ) . F e m a l e genitalianotexamined.

Molecularcharacterization.Nointraspecifichaplotypediversity w a s f o u n d i n t h e a v a i l a b l e s e q u e n c e s ( n = 5 ) . Th el owest overal l m e a n di s t a nc e t o anot her m e m b e r o f thegenusis5.3%toL.tolimafromNevadodelRuiz.BINnumb er:B O L D : AD D7 26 0. D i a g n o s t i c f ixeds t a t e s a n d theirp o si t i o n i n t h e C O I b a r c o d e s e q u e n c e a r e d e p i c t e d inTable2.

Typem a t e r i a l . H o l o t y p e m a l e : C o l o m b i a , Va lle,Tenerife,PáramoLasDomínguez,3600m,29.i.2017,speci mennumber:i1241,sequencepageinBOLD:GWOTU1004- 17,C.Prie-to.Deposit ed i nIC N. Parat ypes,

(8 ♂a nd 1 ♀):1 ♀:C ol om bi a,

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Fig.4.NeighbourJoining(NJ)identificationtreeoffull-lengthbarcodes(658bp)for47speciesofLymanopodausingtheK2P- parametermodel.BIN(BarcodeIndexNumber)assignmentusingBOLDisalsodepicted.

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Table2.Nucleotidesiteswithuniquefixedstates,whichservetoidentifyninespeciesofLymanopodafromColombia.Onlyspecieswiththreeormore individualswereincludedintheanalysis.Red–diagnosticcharacters;orange–partiallydiagnosticcharacters.

Valle,Tenerife,P á r a m o L a s D o m í n g u e z , 3 3 0 0 m , 2 1 . v i i . 2 0 1 3 , specimennumber:i918,sequencepageinBOLD:GWOTR85 9-

16,C . P r i e t o , R C C P ; 3 ♂:C o l o m b i a , Valle,Tenerife,P á r

a m o LasDomínguez,3600m,29.i.2017,specimennumbers:

i1239,

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i1240,i 1 2 4 2 , s e q u e n c e p a g e s i n B O L D : G W O T U 1 0 0 2 - 1 7 , GWOTU1003-17,GW OT U1005-

17, C . Pri e t o, R C C P; 3 ♂C o-

lombia,ValledelCauca,PáramoLasDomínguez,Tenerife,NEP almira0 3 ° 4 5 ’ N , 7 6 ° 0 5 ’ W,3 5 0 0 –

3 6 0 0 m , 2 9 . i . 2 0 1 7 , P B ; 1♂

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Table3.Listofspecimens,localitiesandBOLDaccessionnumbersforeachindividualusedinthemolecularstudy.

Species ProcessID Country Region Lat Lon Elev

Lasiophilazapatoza GWOTU985-17 Colombia Perija 10.3667 –72.95 2200

Coradeschelonis GWOTU974-17 Colombia Perija 10.35 –72.9167 2800

Lymanopodanevada GWOTR880-16 Colombia SierraNevada 11.1 –74.0333 2800

Lymanopodaparamera GWOTR874-16 Colombia SerraniadelPerija 10.35 –72.9167 2800 Lymanopodaparamera GWOTR873-16 Colombia SerraniadelPerija 10.35 –72.9167 2800 Lymanopodaparamera GWOTR872-16 Colombia SerraniadelPerija 10.35 –72.9167 2800

Lymanopodaflammigera GWOTR859-16 Colombia Tenerife 3.68333 –76.1 3600

Lymanopodatolima GWOTR858-16 Colombia PNNLosNevados 4.98333 –75.3333 3200

Lymanopodaflammigera GWOTU1005-17 Colombia Valle,Tenerife 3.68333 –76.1 3600

Lymanopodasamius GWOTR751-16 Colombia Ocana 8.03333 –73.0167 2700

Lymanopodacaucana GWOTR726-16 Colombia Manaure 10.35 –72.9167 1900

Lymanopodaflorenciensis GWOTR735-16 Colombia BotaCaucana 1.51667 –76.3167 1550 Lymanopodaflorenciensis GWOTR734-16 Colombia BotaCaucana 1.51667 –76.3167 1550

Lymanopodaalbocincta GWOTR730-16 Colombia Argelia 2.36667 –77.1833 2700

Lymanopodaalbocincta GWOTR729-16 Colombia Manaure 10.35 –72.9167 2500

Lymanopodaflammigera GWOTU1004-17 Colombia Valle,Tenerife 3.68333 –76.1 3600 Lymanopodaflammigera GWOTU1003-17 Colombia Valle,Tenerife 3.68333 –76.1 3600

Lymanopodamaletera GWOTR723-16 Colombia Villanueva 10.35 –72.9167 2600

Lymanopodaobsoleta GWOTR713-16 Colombia Herran 7.41667 –72.4333 2400

Lymanopodavenosa GBGL8234-12 Perú Puno 1200

Lymanopodaumbratilis GBGL8233-12 Perú Cuzco 1200

Lymanopodashefteli GBGL8232-12 Perú Cuzco 2500

Lymanopodarana GBLN0770-06 Perú Pasco 2500

Lymanopodaprusia GBMIN34609-13 Perú Pasco 2800

Lymanopodanivea GBGL8227-12 Ecuador Napo 2700

Lymanopodanadia GBMIN34613-13 Ecuador Morona-Santiago 2800

Lymanopodamelia GBGL8226-12 Ecuador Tungurahua 3600

Lymanopodamarianna GBGL8225-12 Venezuela Merida 3100

Lymanopodamagna GBMIN34615-13 Perú Molinopampa 2870

Lymanopodalecromi GBGL8224-12 Venezuela Tachira 2700

Lymanopodalabdassp GBMIN34616-13 Colombia Antioquia 2700

Lymanopodainde GBMIN34617-13 Perú Molinopampa 3200

Lymanopodahyagnis GBMIN34618-13 Perú Cuzco 2900

Lymanopodahazelana GBMIN34619-13 Ecuador Loja 3000

Lymanopodaferruginosa GBGL8220-12 Perú Cuzco 2050

Lymanopodaexcisa GBMIN34620-13 Ecuador Loja 3025

Lymanopodaeuopis GBGL8219-12 CostaRica Irazú 2700

Lymanopodaeubagioides GBMIN34621-13 Perú Cuzco 2600

Lymanopodadietzi GBGL8216-12 Venezuela Tachira 2700

Lymanopodaconfusa GBMIN34624-13 Ecuador Zamora-Chinchipe 2100

Lymanopodaaraneola GBGL8213-12 Perú Molinopampa 2870

Lymanopodaapulia GBMIN34627-13 Perú Pasco 2600

Lymanopodaalbomaculata GBGL8211-12 Bolivia Cochabamba 2750

Lymanopodaaffineola GBGL8210-12 Perú Puno 2700

Lymanopodaacraeida GBMIN34630-13 Perú Cuzco 1400

Lymanopodavivienteni GBMIN34605-13 Colombia Guasca 3200

Lymanopodaparamera GWOTR871-16 Colombia SerraniadelPerija 10.35 –72.9167 2800

Lymanopodacasneri GWOTR869-16 Colombia Belmira 6.65 –75.6667 3100

Lymanopodahuilana GWOTR866-16 Colombia Sibundoy 1.13333 –77.0833 3000

Lymanopodapieridina GWOTR758-16 Colombia Calarca 4.46667 –75.55 3200

Lymanopodaionius GWOTR745-16 Colombia Purace 2.16667 –76.3833 3000

Lymanopodaflorenciensis GWOTR737-16 Colombia BotaCaucana 1.51667 –76.3167 1400

Lymanopodaaltis GWOTR733-16 Colombia PNNPurace 2.16667 –76.3833 2900

Lymanopodaflammigera GWOTU1002-17 Colombia Valle,Tenerife 3.68333 –76.1 3600

Lymanopodaobsoleta GWOTR712-16 Colombia Ocana 8.03333 –73.0167 2700

Lymanopodapanacea GWOTR709-16 Colombia BotaCaucana 1.51667 –76.3167 1800

Lymanopodacaeruleata GWOTU968-17 Colombia SierraNevada 10.8833 –74.0167 1500

Lymanopodacaracara GBGL8214-12 Ecuador Napo 3600

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Fig.5.Phylogenetic treeconstructedusingthemaximumlikelihood methodimplementedin thePhyMLprogram(v3.1/3.0 aL RT)andshowingtherelationshipsamongtaxabelongingtoLymanopoda.Numbersrepresentbranchsupportvalues.

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Colombia,Valled e l C a u c a , P á r a m o L a s D o m í n g u e z , Teneri fe,NEPalmira03°45’N,76°05’W,3500–3600m,29.i.2017,P.

Boyerleg.,CEP-

MZUJ;1♂Colombia,ValledelCauca,PáramoLasDomínguez,Tene rife,NEPalmira03°45’N,76°05’W,3500–

3600m,28.i.2017,P.Boyerleg.,prep.genit.498/14.02.2017J.

Lorenc-Brudecka,CEP-MZUJ.

Typelocality.PáramodeLasDominguez(=PandeAzúcar),Te nerife,N E P a l m i r a , Valled e l C a u c a D e p a r t m e n t , 0 3 ° 4 5 ’ N , 76°05’W,3500–3600ma.s.l.

Etymology.Thespecificepithet“flammigera”isthenomina- tivefemininesingularof“flammiger”fromthelatin“flamma”

(=flame)and-iger(gero)

(=tocarry,tobear),inreferencetotheintenseorange- yellowcolourofthemalesofthisbutterfly.

Bionomics.Malespatrolat1–

2mabovethegroundinthec l o u d f o r e s t –

p á r a m o e c o t o n e . M a l e s p a t r o l a r o u n d middayalongt hesunnyedgesofpaths.Theimmaturestag-

esandlarvalfoodplantsareunknownbutarepresumedtobeChu squeabamboo,asisthecasewithotherspeciesofLymanopo da,commoninthecollectingarea.

Distribution.Thisspeciesisknownonlyfromthetypeloc ality,P á r a m o d e L a s D o m í n g u e z ( P a n d e A z u c a r i n somemaps),anisolatedmassifsituatedwestofthemai nColombianCentralCordillerarangeoftheAndes.Itoccursinthepár amograsslandat3300–3600m,justabovetim-

berline.AdultswerecollectedinJanuaryandJuly.

Speciesdelimitationbasedonbarcodeanalysis

AN J t r e e w a s g e n e r a t e d f o r 4 7 s p e c i e s a n d 7 9 i n d i -

vidualsofLymanopoda.Whendiscrepancies between t heD N A -

b a s e d a n d s t a n d a r d t a x o n o m y w e r e f o u n d , t h e spe cimenw a s e x a m i n e d t o c o nfirmt h a t i t s m o r p h o l o g i - calidentificationwascorrect,andthealignmentandtracefiles werecarefullyre-examined.Itwasfoundthat47mor-

phospecieswe re a ss i g ne dt o4 4 B I N s ( F i g .4 ), t h er e f or e showinga94%ofcongruencebetweenmorphospeciesandBIN s.T h e m o r p h o s p e c i e s L . h y a g n i s ,L . u m b r a t i l i s a n d

L.shefteliwereplacedintheNJtreeunderthesameBINcode duetothelowgeneticdivergenceof0.39%between

L.hyagnisandL.umbratilis,and1.9%betweenL.shefteliandL.

hyagnis.Similarly,thegeneticdivergencebetweenthethem orphologicallyverydivergentL.caeruleataand

L.caucanais0.77%

(Table1).However,inallthecases,theidentificationofanun knownspecimenbymatchingitssequencetothoseinthereference libraryledtocorrectre-

sults.Nucleotidesiteswithunique fixedstatesthatwe reusedtoidentifyninespecies(thoserepresentedbyatleastthr eespecimensinourdataset)ofLymanopodafromCo- lombiaarecomparedinTable2.

ThespeciesofLymanopodaexaminedhaveameanin- traspecificgeneticdistanceof0.05%

(n=78comparisonsofbarcodes>600bp).Maximumintraspec ificdivergencewas0.77%.Themeaninterspec ificgene ticdist ance was 9.60%

(n=2848comparisonsofbarcodes>600bp).Max-

imuminterspecificdivergencewas13.37%andminimuminte rspecificdistancewas0.39%.

Lymanopodaphylogeny

AphylogenetictreewasconstructedusingtheMLmeth- odfor47speciesofLymanopoda,including17fromCo- lombiaand30otherswhoseCOIsequenceswereavailable

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inGenBank(Fig.5),outof~

65known,whichmakesup70%ofallknownspecies.Thetreepr esentsfourmainclad-

es,oneofwhichiscalledhereforconvenience“obsoleta”with 14speciesincludingthetwospecies,L.florenciaensisSalazar,He nao&Vargas,2004andL.maleteraAdams&Bernard,1 97 9 ,n ot s eq ue nc e db e fo r e ,t he “ionius”c l a d e with17spe cies,the“caucana”cladewithfivespeciesand“tolima”cladew itheightspecies.Thelatterissubdividedintotwoclades,oneo fwhichincludestwospeciesnotse-

quencedbefore,L.nevadaKrüger,1924andL.parameraA dams&Bernard, 1979, whereast he ot her cont ai nss ix speciesincludingL.flammigerasp.n.andtwootherspe -

ciesnotincludedinthegenericphylogenyproducedpre -

viously(Casner&Pyrcz,2010),L.tolimaandL.casneri Pyrcz&Clavijo,2016,thelatter,however,sequencedby Marínetal.

(2017).Theresolutionofthiscladeislowandpresentsapolyto my,thereforethepositionofthenewspe-

ciesrelativetootherfivespeciesisnotestablished.

DISCUSSIO N

Colourpattern s

Thenewspeciesisremarkablefirstofallbecauseofitsun usualgol den-

yel l ow co l o ur of m a l e s, uni que no t o nl y amongothe rcongenersbutalsowithintheentirespecies-

richs u b t r i b e P r o n o p h i l i n a ( o v e r 6 5 0 s p e c i e s ) , a n d a rgu-

ablyamongallneotropicalandevenworldwideSatyrinae.Thee volutionarybasisof thisoutstandingcolourationi sunknownbutthehypot hesi sthat thiscolouringis s ome-

howrel at ed t om i m i c r y,se em su nl i k el y.Thisi sb ec au se theSulphurColiasdimeraDoubleday,1847,whichi sthepotentialmodel,althoughgenerally very comm onintheColombianpáramosandprobablyobnoxious,hasnotb eendetectedintheregionwhereL.flammigerasp.n.occur s.OtherrelatedColombianspecies,suchasL.huilana,L.to- lima,L.zebraPyrcz&Rodríguez,2007,L.casneriandL.meli aWeymer,1911,arepredominantlywhiteorblackandwhite,whic hiscertainlyassociatedwiththermoregulationandt h e l i m i t e d s o l a r r a d i a t i o n a t h i g h a l t i t u d e s , a n d t h e highera bsorptionofUV.Itcouldeventuallyalsoprovetobethecasef orL.flammigeraalthoughtheopticalqualitiesofitswingpigmen tsandscaleshavenotbeeninvestigatedsofar.Itisworthpointingo utthatthereareseveralsimilar-

lypigmentedspeciesofskippers(Hesperiidae)inthehightropi calandtemperate Andeswithinthegenera Zalomes Bell,WahydraSteinhauser,HylephilaBillberg,andoneyetundesc ribedspeciesofRactaEvans.

Itishoweverpuzzlingwhysuchunusualcolourpatternsevolv edinjustoneisolatedareawhereasthroughoutthe no rtherna n d c e n t r a l Andes m o s t p á r a m o s p e c i e s o f Ly-

manopodaarepredominantlywhite.Ontheotherhand,itis

t ruet hatt hegenus Lymanopodai s part i cul arl y pl as t i c phenotypicallyandanumberofspeciesoccurringincloudforest sortheforest-

páramoecotonehavecolourpatternsthatareunusualfort hesubfamilySatyrinae,forexample,theblueL.hazelana Brown,1943,L.samiusWestwood,1851a n d L.cinnaWe stwood,1889,greenp a t c h e d L . mariannaStaudinger,18 97orredL.inauditaPyrcz,2010.Someofthesecolourpatterns arealmostcertainlydueto

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mimeticrelationships,anissuecurrentlybeinginvestigat- ed(Pyrcz,inprep.).

Barcoding

Thisstudyprovidesaninitialassessmentoftheuseful- nessofDNAbarcodinginLymanopoda.TheNJtreeanaly- sisyieldedhighpercentageofcorrectidentificationsinthegenus Lymanopoda.Inthetree,94%ofthemorphospeciesusedinthisst udyformeddistinctcladesandwereassignedaB a r c o d e I n d e x N u m b e r ( B I N ) m a t c h i n g p e r f e c t l y t h e morphol ogybasedidentifications.In6%ofthecases,morethanonemorpho speciessharesaBINnumberwithotherspecies.Theseca sesincludefivespeciesinthisstudy:L.hyagnis,L.umbratili s,L.shefteliclustertogetherandhavethesameBINnumber;and L.caeruleataandL.caucanaalsohavethesameBINnumbe r.Theformerthreespeciesbelongtoacomplexgroupofmorpholo gicallysimilartaxaoccurringallopatricallyinparallelvalleysinth eMadredeDiosupperbasi ni nsouthernP eruandnorth ernB ol ivi a, whoserelationshipsarestillnotfullyunderstood,an dtheirseparatespecificstatusisyettobeconfirmedbymorethor- oughtaxonomicstudiesinvolvingtheirspatial,geographicandaltit udinaldistributionpatterns.L.caeruleataand

L.c a u c a n a a r e a l l o p a t r i c s p e c i e s , m o r p h o l o g i c a l l y e a s -

ilyseparablebytheirpredominantlyblue (L.caeruleat a)andbrown(L.caucana)wingcolourpatternsandgenit alcharacters,sotheirseparatespecificstatusisstronglysup- ported.OurresultsconfirmthatDNAbarcodingisahighlyefficient methodforidentifyingspecies inthesubfamilySaty rinae,aspointedoutinanotherrecentstudyonhighAnde anbutterflies(Marínetal.,2017).

Phylogeny

ThecladogrambasedontheCOImarkerproducedfo r47specieshastobeconsideredascomplementaryrelativetopre viousstudiesasittakesintoconsiderationonlyonemarker, comparedto40speciesand5molecularmarkers(Casner

&Pyrcz,2010).We,however,chosetouseonlytheCOIm arkerbecauseoneofthekeyissuesofthisstudywastoinvestigat etherobustnessofbarcodingrelativetomorphologicaltr aitsinevaluatingrelationshipswithinthegenusLymanopoda, inparticular,betweenhypotheticallycloselyrelatedtaxa.

Itisinteresting,fromthisperspective,topo i nt ou t t h a t , r eg ar di ng t h e su bdi vi si o n o f t h eg e n u s intomainmonophyleti cgroupsand,inparticular,thebasalpositiono f t h e “caucana”c l a d e c o m p r i s i n g 5 s p e c i e s , theresultsarehighl ycongruent withpreviousmolecular(Casner&Pyrcz, 2010)andmorphologicalphylogenetic hypotheses(Py rcz,2001).ThepositionofL.prusia,Heim-

lich,1973,asasistertotheremainingspeciesofLymanop- odais,however,notconfirmed.

The“tolimaclade”,with6speciesinCasner&Pyrcz’spap er,ishererestrictedto4species,twoofwhichwerenotpreviouslyexa mined,L.huilanaandL.flammigerasp.n.Thisw e l l s u p p o r t e d c l a d e i n c l u d e s a l l t h e h i g h a l t i t u d e páramo species,examinedsofar,distributedfromnorth- centralColombia(Belmira)toEcuador.Also,alloftheses peciesshareanumberofmorphologicalsynapomorphies,whichsu pportitsmonophyly.Inthisrespect,thePeruvian

species,L . i n d e P y r c z , 2 0 0 4 a n d L . e u b a g i o i d e s B u t l e r,1873,excludedfromthisclade,standapart,andthei rpo-

sitionwithinthiscladesuggestedoriginallybyCasner&

Pyrcz(op.cit.)shouldbereconsidered. Importantly,t wowhitepáramospecies,L.nevadaandL.paramera,fou ndinisolatedrangesinnorthernColombia,wereincludedint hemolecularanalysisforthefirsttime.Althoughtheysu-

perficiallyresemblethe speci es inthe“tolima”cl ade by beingpredominantlywhite,theywereplacedinaseparateclade, eveniftheystilloccurinthelargerunitcomprisingthe“tolima”

cladeandnotintheothertwolargeclades, “excisa”and“

obsoleta”.

Byc o m b i n i n g m o l e c u l a r a n d m o r p h o l o g i c a l d a t a i t i s possibletodeterminetheclosestrelativesof L.fl ammig-

erasp.n.withinthe“tolima”clade.COIbasedanalysisisinconc lusiveinthisrespectinshowingapolytomy.Com-

parisonsofmalegenitaliashow,however,thatL.flammig- era,L.casneriandL.tolimashareauniquesynapomorphy,abifurca te,dorso-

ventrallyflattened,prominentrounded superuncus.In L.huilanaandL.hazelanathesuperuncusisconsiderablys mallerandnotbifurcatedeveniftwolat-

erallobesarenoticeable.Othercharactersarelessevident,althoug hthevalvaeofL.casneriandL.tolimaaremoresimilar,b eingshortwithasingleprominentapicaltooth,wherea sthevalvaeofL.flammigerasp.n.arenarrowerinthemiddleand muchlonger,lookinginthisrespectmorelikethoseofL.huil ana.InL.melia,thesisterspeciesofL.tolimaaccordingtoCasner

&Pyrcz’s(op.cit.)phylogeny,thesuperuncusisshortandsingle.

Thesedatahaveimpor-

tantphylogeographicalimplications.L.tolimadivergedinthefi rstplacefromL.huilanaeventhoughthereisacon-

tinuityofpáramohabitatsbetween theareasin Quindí o,TolimaandValledelCaucaintheCentralCordillerawithth oseinCaucaandNariñofurthersouthwhereL.huilanaoccur s.Ontheotherhand,therearecurrentlynoappropri-

atepáramohabitatsover200kmbetweenQuindioandthePáramo deBelmirainAntioquiawhereL.casneriisfound.Apparentlysome morecomplexunderlyingpaleoecologi-

calprocesseshaveresultedintheshapingofpresentda ydistributionso fp ár am o Lymanopodasp ec i e s i nt h i sp a rt ofColombia.

Finalconsiderations

Thisstudyhighlightstwoimportantfacts.Itisconfirmedthatgeni talia,inparticularthoseofmales,areextremelyvalua blen o t o n l y i n a l p h a -

t a x o n o m y b u t a l s o p h y l o g e -

netically.Here ac om pa r at i v ea n al ys i s r efinessom ed at a obtainedusingmoleculartools.Ofcourse,notinallta xaaremalegenitaliaas informative,whichdepends mostlyonthenum berof m od ificationsl eadi ngt o thee vol ut i on ofn o t i c e a b l e p h e n o t y p i c a l t r a i t s e v e n i n c l o s e l y r e l a t e d groupsoftaxa.Int he genus Lyman opodasuch t rait sare appreciable.Secondly,ourstudyc onfirmstheusefulness oftheCOImarkerinspeciesdefinit ionaswellasinphy-

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logeneticconsiderations,arolethathasbeenquestione d.Here,COIsupportdataon17speciesofColombianLyma- nopodahelpedrefinethephylogenyofthegenus,andi scongruenti n m o s t a s p e c t s w i t h t h e p r e v i o u s l y p r o p o s e d arrangementbasedonfivemarkers.Inotherw ords,COI

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doesw o r k a t l e a s t i n t h e g e n u s Lymanopoda,e v e n i f i n someothertaxa ofLepidoptera t his maynotnecessar ily bet h e c a s e . Thi s st u d y e x p a n d e d o u r k n o w l e d g e o n t h e evolutionofthegenusLymanopodabyaddingsevenmor especiestoitsphylogeny.Dataforseveralkeyspeciesare howeverstillmissing,inColombiainparticularforL.mi- rabilis(Staudinger),ahighpáramospecieswithunusu alextremelyelongatedwingsandatypicalgenitaliafromtheso uthernpartoftheCordilleraOriental,andL.melendezaAdam sfr om t h e S i e r r a d el C o c u y t h a t ha s s om e r e s e m - blanceinbothgenitaliaandcolourpatternstotheVenezue- lanL.mariannaStaudinger,knownsofaronlyfromt heholotype.

ACKNOWLEDGEMENTS.Forhelpandsupportinmanywaysweackno wledgeA.Hausmann(BavarianStateCollectionofZool- ogy,Munich,Germany)andC.UribeOrtegaandL.C.Gutierrez(Uni versidadd e l A t l á n t i c o , B a r r a n q u i l l a , C o l o m b i a ) . T h i s r e -

searchwassupportedbyfundsfromtheGeorgForsterResearchFell owshipProgramoftheAlexander-

vonHumboldtFoundation(Bonn),theFederalMinistryforEduca tionandResearch(Ger-many),theVice-

RectorateforResearch oftheUniversidad delAtlántico,Colo mbiaunderdecisionnumber3247of 12thJune2015,andbyan internalgrantoftheInstituteofZoologyandBio-

medicalResearchoftheJagiellonianUniversity,KZDS006320.

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ReceivedJanuary11,2018;revisedandacceptedJune8,2018 PublishedonlineAugust16,2018