phylogenetic relationships and historical biogeography of tribes … · (lepidoptera: nymphalidae)...

Biological Journal of the Linnean Society

, 2005,

86

, 227–251. With 5 figures

© 2005 The Linnean Society of London,

Biological Journal of the Linnean Society,

2005,

86

, 227–251

227

Blackwell Science, LtdOxford, UKBIJBiological Journal of the Linnean Society0024-4066The Linnean Society of London, 2005? 2005862227251Original Article

PHYLOGENY OF NYMPHALINAEN. WAHLBERG

ET AL

.

*Corresponding author. E-mail: [email protected]

Phylogenetic relationships and historical biogeography of tribes and genera in the subfamily Nymphalinae (Lepidoptera: Nymphalidae)

NIKLAS WAHLBERG

1

*, ANDREW V. Z. BROWER

2

and SÖREN NYLIN

1

1

Department of Zoology, Stockholm University, S-106 91 Stockholm, Sweden

2

Department of Zoology, Oregon State University, Corvallis, Oregon 97331–2907, USA

Received 10 January 2004; accepted for publication 12 November 2004

We infer for the first time the phylogenetic relationships of genera and tribes in the ecologically and evolutionarilywell-studied subfamily Nymphalinae using DNA sequence data from three genes: 1450 bp of cytochrome oxidasesubunit I (COI) (in the mitochondrial genome), 1077 bp of elongation factor 1-alpha (EF1-

a

) and 400–403 bp of

wing-less

(both in the nuclear genome). We explore the influence of each gene region on the support given to each node ofthe most parsimonious tree derived from a combined analysis of all three genes using Partitioned Bremer Support.We also explore the influence of assuming equal weights for all characters in the combined analysis by investigatingthe stability of clades to different transition/transversion weighting schemes. We find many strongly supported andstable clades in the Nymphalinae. We are also able to identify ‘rogue’ taxa whose positions are weakly supported (thedifferent gene regions are in conflict with each other) and unstable. Our main conclusions are: (1) the tribe Coeinias currently constituted is untenable, and

Smyrna, Colobura

and

Tigridia

are part of Nymphalini; (2) ‘Kallimini’ isparaphyletic with regard to Melitaeini and should be split into three tribes: Kallimini

s.s.

, Junoniini and Victorinini;(3) Junoniini, Victorinini, Melitaeini and the newly circumscribed Nymphalini are strongly supported monophyleticgroups, and (4)

Precis

and

Junonia

are not synonymous or even sister groups. The species

Junonia coenia

, a modelsystem in developmental biology, clearly belongs in the genus

Junonia

. A dispersal-vicariance analysis suggests thatdispersal has had a major effect on the distributions of extant species, and three biotic regions are identified as beingcentres of diversification of three major clades: the Palaearctic for the

Nymphalis

-group, the Afrotropics for Junoniiniand the Nearctic for Melitaeini. © 2005 The Linnean Society of London,

Biological Journal of the Linnean Society

,2005,

86

, 227–251.

ADDITIONAL KEYWORDS:

DIVA – molecular phylogeny – Partitioned Bremer Support – sensitivity analysis.

INTRODUCTION

Butterflies belonging to the currently recognized sub-family Nymphalinae (Wahlberg, Weingartner & Nylin,2003b) have contributed extensively to our knowledgeof ecological and evolutionary processes, from hybridzones and ring-species (Forbes, 1928; Silberglied,1984; Dasmahaptra

et al

., 2002; Austin

et al

., 2003),metapopulation dynamics (Hanski, 1999) and evolu-tionary developmental biology (Carroll

et al

., 1994;Brakefield

et al

., 1996), to insect–plant interactions(Singer, 1971; Nylin, 1988; Janz, Nylin & Nyblom,

2001; Wahlberg, 2001). Despite the long-standinginterest in these butterflies, the phylogenetic relation-ships among the various tribes and genera haveremained remarkably obscure. Improving our under-standing of the phylogenetic resolution of such scien-tifically popular taxa should be a high priority, so thatthis abundance of knowledge can be placed in an evo-lutionary framework.

Since Nymphalinae is the type subfamily of thediverse family Nymphalidae, its delineation hasenjoyed a dynamic history, as various authors haveconsidered diverse subsets of tribes and genera to rep-resent ‘typical nymphalids’. This confusion has led toseveral competing classification schemes based on dif-ferent data sets (Ackery, 1988; Harvey, 1991; Kuznet-

mailto:[email protected]

228

N. WAHLBERG

ET AL

.



2005,

86

, 227–251

zov & Stekolnikov, 2001; Wahlberg

et al

., 2003b). Thehigher systematics of Nymphalidae is still in a state offlux and the delineation of Nymphalinae has not yetreached stability, though there is a growing consensusthat the classification of Harvey (1991) (based on theclassification of Müller, 1886), with the addition of thetribe Coeini (= Coloburini of authors), currently pro-vides the most natural definition of the subfamily(Brower, 2000; Wahlberg

et al

., 2003b; Freitas &Brown, 2004). This concept of Nymphalinae comprisesa monophyletic group that includes the supposedlywell-defined tribes Nymphalini, Coeini, Melitaeiniand Kallimini. It is this hypothesis of nymphalinerelationships that we take as our point of departure inour analysis and discussion.

Ehrlich’s (1958) influential paper on the classifica-tion of butterflies included a much broader concept ofNymphalinae, which was based mainly upon symple-siomorphic and homoplastic characters and appearedto comprise those taxa that do not fall into any of hismore restricted and homogeneous nymphalid subfam-ilies (Danainae, Ithomiinae, Satyrinae, Morphinae,Calinaginae, Charaxinae, Acraeinae). The taxa in Ehr-lich’s Nymphalinae are today considered to representseveral different subfamilies, including Heliconiinae,Limenitidinae, Biblidinae and Apaturinae. Severalsubsequent authors have used much the same groupof tribes and genera to represent the ‘core nymphalids’(e.g. Ackery, 1984; Scott, 1985; Scott & Wright, 1990),though often splitting off some components as distinctsubfamilies. At the other extreme, some authors havedelineated Nymphalinae in a very narrow sense, toinclude only the tribes Nymphalini and Kallimini(Ackery, 1988) or Nymphalini and Melitaeini (Clark,1948).

All of the above classifications have suffered from alack of clearly described morphological synapomor-phies that diagnose the various circumscriptions. Har-vey (1991) proposed a classification of the familyNymphalidae based on a set of larval characters, thatwas accepted by many authors (e.g. Ackery

et al.

,1999). He placed the tribes Nymphalini, Kallimini andMelitaeini together, based on the arrangement ofspines on the larvae, but was unable to resolve therelationships among them due to character conflictwithin the subfamily. Harvey (1991) placed the genus

Amnosia

in the tribe Kallimini, but Wahlberg

et al

.(2003b) showed that

Amnosia

does not belong inNymphalinae, but in Cyrestinae along with othermembers of Pseudergolini (with which

Amnosia

wasaffiliated prior to being moved to Kallimini (Ackery,1988).

Most historical classifications of the Nymphalidaehave been intuitive rather than the product of rigor-ous phylogenetic analysis of character state distribu-tions formalized in a data matrix. More recently,

however, several cladistic analyses have been pub-lished, based on either morphology (DeVries

et al.

,1985; de Jong

et al.

, 1996; Penz & Peggie, 2003; Freitas& Brown, 2004) or DNA sequences (Weller

et al.

, 1996;Brower, 2000; Wahlberg

et al

., 2003b). Three of thesestudies (Brower, 2000; Wahlberg

et al

., 2003b; Freitas& Brown, 2004) sampled enough representatives ofNymphalidae to provide evidence bearing upon themonophyly and circumscription of the Nymphalinae.In all three, sampled taxa belonging to Nymphalini,Coeini, Kallimini and Melitaeini form a monophyleticgroup, with Coeini as the sister group to Nymphalini.Freitas & Brown (2004) and Wahlberg

et al

. (2003b)suggest an association of Kallimini with Melitaeini,while Brower’s (2000) study has a basal, paraphyleticKallimini, with regard to Melitaeini and Nymphalini

+

Coeini. The sister group to the Nymphalinae remainsin doubt, with Freitas & Brown (2004) proposing Hel-iconiinae, Brower (2000) suggesting Biblidinae

+

Apa-turinae and Wahlberg

et al

.’s (2003b) data implyingApaturinae.

Within Nymphalinae, several taxa have receivedattention from systematists in recent years. The rela-tionships among genera and species groups have beeninvestigated in Melitaeini (Kons, 2000; Wahlberg &Zimmermann, 2000) and Nymphalini (Nylin

et al

.,2001; Wahlberg & Nylin, 2003) using both molecularand morphological data. In Melitaeini, it has becomeclear that Harvey’s (1991) proposed division into threesubtribes (Euphydryina, Melitaeina, Phyciodina) isnot satisfactory. Wahlberg & Zimmermann (2000),based on mtDNA, found that the

Euphydryas

-,

Meli-taea-, Chlosyne-

and

Phyciodes-

groups of species andgenera were of equal status. Kons (2000), using mor-phology, also found these four major groups andadditionally described a fifth group containing

Gna-thotriche

species. However, the relationships of thefour major groups are in conflict between these twostudies. Wahlberg and Zimmermann inferred the

Mel-itaea

-group to be sister to the

Chlosyne-

group,whereas Kons found Phyciodina and

Gnathotriche

tobe sister to the

Chlosyne

-group.The two studies on Nymphalini (Nylin

et al

., 2001;Wahlberg & Nylin, 2003) have established the mono-phyly of the the

Nymphalis

-group of genera (i.e.

Aglais, Nymphalis

and

Polygonia

) and the close rela-tionship between

Araschnia, Mynes

and

Symbrenthia

,but otherwise the relationships of genera withinNymphalini remain unclear. A further recent morpho-logical study of

Symbrenthia, Mynes

and

Araschnia

(Fric, Konvicka & Zrzav

y

, 2004) suggests that

Mynes

is nested within

Symbrenthia

.In addition to these tribal-level studies, species-

level phylogenetic studies have been done for the gen-era

Euphydryas

(Zimmermann, Wahlberg & Desci-mon, 2000),

Chlosyne

(Kons, 2000),

Hypanartia

PHYLOGENY OF NYMPHALINAE

229



2005,

86

, 227–251

(Willmott, Hall & Lamas, 2001),

Anartia

(Blum, Ber-mingham & Dasmahapatra, 2003),

Phyciodes

(Wahl-berg, Oliveira & Scott, 2003a) and

Symbrenthia

,

Mynes

and

Araschnia

(Fric

et al

., 2004). Of these, onlyKons (2000), Wahlberg

et al

. (2003a) and Fric

et al

.(2004) included sufficiently extensive outgroup sam-pling to test the monophyly of the genus being studied.

From this brief review of the current state ofnymphaline systematics, it is clear that many ques-tions remain unanswered. The relationships amonggenera in Kallimini and Coeini have never been inves-tigated, and relationships within Melitaeini andNymphalini are still contentious. In this study we testthe monophyly of the most recent definition of Nymph-alinae, comprising Nymphalini, Coeini, Kallimini andMelitaeini (Wahlberg

et al

., 2003b), and endeavour toresolve the relationships among the tribes and generawithin the subfamily. In the current circumscription,the subfamily Nymphalinae comprises about 496 spe-cies in 56 genera. We have studied the relationships ofrepresentative species belonging to the subfamily withDNA sequences from three gene regions. These arecytochrome oxidase subunit I (COI) from the mito-chondrial genome, and elongation factor-1

a

(EF1-

a

)and

wingless

from the nuclear genome. Based on ourresults, we investigate the biogeography of the groupusing dispersal-vicariance analysis (Ronquist, 1997).The investigation is intended to identify broad biogeo-graphical patterns for closer inspection at a later date.

MATERIAL AND METHODS

We sampled as many species as possible from almostall of the genera belonging to Nymphalinae, a total of161 species in 49 genera. Of the seven missing genera,six are in Melitaeini and one is in Coeini. In addition,we sampled 28 outgroup species, representing all sub-families of the nymphaline clade (

sensu

Wahlberg

et al

., 2003b), i.e. Cyrestinae, Biblidinae and Apaturi-nae, each of which may be the sister group to Nymph-alinae (see Wahlberg

et al

., 2003b). We also included aspecimen of Heliconiinae and Limenitidinae, whichbelong to the putative sister clade to the nymphalineclade. The species sampled and their collection locali-ties are listed in Appendix 1.

We extracted DNA mainly from one or two legs offreshly frozen or dried butterflies using QIAgen’sDNEasy extraction kit, although some specimenswere extracted following the protocol of Brower (1994).The spread voucher specimens can be viewed at http://www.zoologi.su.se/research/wahlberg/. For each spec-imen we sequenced 1450 bp of COI, 1077 bp of EF1-

a

and 400–403 bp of the

wingless

gene. Primers for COIwere taken from Wahlberg & Zimmermann (2000), forEF1-

a

from Monteiro & Pierce (2001) and for

wingless

from Brower & DeSalle (1998). We performed all PCRs

in a 20

m

L reaction volume. The cycling profile for COIand

wingless

was 95

∞

C for 5 min, 35 cycles of 94

∞

C for30 s, 47

∞

C for 30 s, 72

∞

C for 1 min 30 s and a finalextension period of 72

∞

C for 10 min. The cycling pro-file for EF1-

a

was 95

∞

C for 7 min, 35 cycles of 95

∞

Cfor 1 min, 55

∞

C for 1 min, 72

∞

C for 2 min and a finalextension period of 72

∞

C for 10 min. For all threegenes, the PCR primers were also used for sequencing.

In addition, we developed two internal primers forsequencing: (EFmid 5

¢-

CAA TAC CRC CRA TTTTGT

-

3

¢

) for EF1-

a

and (Patty 5

¢-

ACW GTW GGWGGA TTA ACW GG

-

3

¢

) for COI. Sequencing was donewith a Beckman-Coulter CEQ2000 or CEQ8000 capil-lary sequencer (Bromma, Sweden). We checked theresulting chromatograms using BioEdit (Hall, 1999)and aligned the sequences by eye. Clear heterozygouspositions in the nuclear genes (chromatogram peaksalmost or exactly equal) were coded according to theIUPAC ambiguity codes, but were treated as missingcharacters in further analyses. The sequences havebeen submitted to GenBank (Accession numbers inAppendix 1).

We searched for the most parsimonious cladogramsfrom the equally weighted and unordered data matrixconsisting of 189 taxa using a heuristic search algo-rithm in NONA 2.0 (Goloboff, 1998) via WINCLADA1.00.08 (Nixon, 2002). The heuristic searches wereconducted with 1000 random addition replicates usingTBR branch swapping with ten trees held during eachstep and a final swapping to completion. We did thisfor each gene separately and for all three genes com-bined. Trees were rooted with

Heliconius

for display.For the separate analyses, we evaluated clade supportusing bootstrap with 100 pseudoreplicates. It is nowwidely recognized that assessing incongruence amongdata partitions is much more complex than measuringwith a simple all-or-nothing significance test (Farris

et al

., 1994; DeSalle & Brower, 1997; Miller et al.,1997; Darlu & Lecointre, 2002). We have thus chosento analyse the three gene regions as a single data set,and have assessed the impact of each gene region onthe support values of each node using PartitionedBremer Support (PBS) analyses (Baker & DeSalle,1997; Baker et al., 1998).

We evaluated the character support for the clades inthe resulting cladograms using Bremer support (BS)(Bremer, 1988, 1994). We calculated BS and PBSvalues using anticonstraints in PAUP* 4.0b10 forWindows (Swofford, 2001). As in previous studies(Wahlberg & Nylin, 2003; Wahlberg et al., 2003b), werefer to the support values as giving weak, moderate,good or strong support when discussing our results.We define ‘weak support’ as BS values of 1–2 (gener-ally corresponding to bootstrap values <50%–63%),‘moderate support’ as values between 3 and 5 (boot-strap values 64%-75%), ‘good support’ as values

http://www.zoologi.su.se/research/wahlberg

230 N. WAHLBERG ET AL.

© 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 86, 227–251

between 6 and 10 (bootstrap values 76%-88%) and‘strong support’ as values >10 (bootstrap values 89%-100%). We strongly endorse BS values over bootstrapvalues because they are a parameter of the datarather than an estimate based on manipulated sub-samples of the data, and have no upper bound as sup-port for a given clade increases.

We evaluated the stability (sensu Wheeler, 1995;Judd, 1998; Giribet, 2003) of clades inferred for theequally weighted data set to different character-statetransformation weighting assumptions under parsi-mony searches using PAUP*. We weighted transver-sions 2, 3, 5, 7 and 10 times transitions for thecombined data set. Such sensitivity analyses may helpidentify potential instances of long branch attraction(Giribet, 2003), and can provide a valuable heuristictool to guide subsequent sampling strategies forrefinement of the current hypothesis. We refer toclades that are recovered under all the tested weight-ing schemes as stable.

We investigated the historical biogeography of thesubfamily using dispersal-vicariance analysis (Ron-quist, 1997) as implemented in DIVA (Ronquist,1996). Species distributions were recorded at the levelof zoological biomes, i.e. Nearctic, Neotropical,Afrotropical, Palaearctic, Oriental and Australasian.Clades for which component species had identical dis-tributions were collapsed into a single terminal. Themaximum number of ancestral areas was either notconstrained or restricted to two.

RESULTS

CHARACTERISTICS OF THE DATA SETS

The total combined data set consisted of 2935 nucle-otides, of which 12 positions were coded as gaps insome taxa. All inferred gaps were in the wingless dataset and represent indel events of whole codons. Theseinclude an inferred codon insertion in all Melitaeinisequences (as noted in Brower, 2000), an inferredcodon insertion in Tigridia acesta, an inferred codoninsertion in Rhinopalpa polynice and an inferredcodon deletion in the three species of Aglais (as notedin Nylin et al., 2001). All inferred indel events have

occurred between positions 91 and 137 in the Coloburadirce sequence (GenBank accession numberAY090162) and were easily detected when aligning byeye (flanking regions were relatively conserved). Gapswere treated as a ‘fifth base pair’ in all phylogeneticanalyses. The basic statistics for the three generegions are given in Table 1. All three gene regionsshowed an ample amount of variation, though EF1-awas less variable than the other two genes. The twonuclear genes show about equal base frequencies,while COI has a high AT bias, in accordance with allprevious publications on this gene in insects.

GENERAL PHYLOGENETIC PATTERNS

When analysed as separate data partitions, none ofthe three gene regions recovered the subfamilyNymphalinae as a monophyletic entity (data notshown). There are, however, several phylogenetic com-ponents common to the separate hypotheses: Meli-taeini and Nymphalini (as delineated below) aremonophyletic, the genera Junonia, Precis, Hypolim-nas, Yoma, Salamis and Protogoniomorpha togetherform a monophyletic group, and the Nymphalis-groupof species (see Wahlberg & Nylin, 2003) is monophyl-etic.

Combining the three data sets yields eight trees oflength 17547 steps (CI = 0.13, RI = 0.58), the strictconsensus of which gives a much clearer picture ofthe relationships among the various groups inNymphalinae (Figs 1–4). The subfamily as currentlydelimited is inferred to be polyphyletic, with thegenera Historis and Baeotus branching off close tothe base of the larger nymphaline clade (sensu Wahl-berg et al., 2003b). The monophyly of Nymphalinaewithout Historis and Baeotus receives moderate sup-port. Nymphalini and Melitaeini form monophyleticgroups, while Coeini and Kallimini are poly- andparaphyletic, respectively. Three genera tradition-ally placed in Coeini (Colobura, Tigridia andSmyrna) are associated with Nymphalini with goodsupport, while the two other sampled genera, His-toris and Baeotus, are outside Nymphalinae withweak support. Kallimini + Melitaeini has moderatesupport, but the kallimine taxa form a paraphyletic

Table 1. The basic statistics for the three molecular data sets in 174 species of Nymphalidae

Gene No. of sites No. variable No. informative

Empirical base frequencies (%)

T C A G

COI 1450 775 614 39.9 14.7 31.9 13.6EF1-a 1077 469 364 22.6 28.1 25.9 23.4wingless 412 238 192 21.0 26.2 24.9 27.9

PHYLOGENY OF NYMPHALINAE 231


Figure 1. Summary of strict consensus of eight trees found for the combined data set when all changes weighted equally(length = 17547, CI = 0.13, RI = 0.58), pruned to show only genera. For unpruned trees, see Figs 2–4. Numbers above thebranches are Bremer support values and numbers below are Partitioned Bremer Support values for the COI, EF1-a andwingless data partitions, respectively. Thickened branches are stable to changes in weighting schemes (transversionsweighted 1, 2, 3, 5, 7 and 10 times transitions).

HeliconiusAdelphaMarpesia chironMarpesia orsilochusChersonesiaCyrestisHistoris odiusHistoris acherontaBaeotus japetusBaeotus deucalionBaeotus aeilusBaeotus beotusDichorragiaStibochionaAmnosiaPseudergolisApaturaMimathymaTimelaeaEulaceuraAsterocampaChitoriaSeveniaDynamineHamadryasCatonepheleMysceliaPanaceaCallicoreNicaAriadneEurytelaBybliaMesoxanthaColoburaTigridiaSmyrnaAntanartiaSymbrenthiaAraschniaMynesHypanartiaVanessaAglaisNymphalisPolygoniaKallimoidesSiproetaNapeoclesMetamorphaAnartiaRhinopalpaVanessulaHypolimnasPrecisJunoniaSalamisYomaProtogoniomorphaDoleschalliaKallimaCatacropteraMallikaEuphydryasPoladryasMicrotiaChlosyneMelitaeaGnathotricheHigginsiusMaziaTegosaPhyciodesAnthanassaTelenassaCastiliaEresiaJanatella

Cyrestini

Coeini

Pseudergolini

Apaturinae

Biblidinae

Nymphalini

Victorinini

Junoniini

Kallimini

Melitaeini

3213.5,17.1,1.4

323.9,-6.0,-14.9

1

18.5,-2.6,-4.9

177.2,9.1,0.7

3127.7,11.4,-8.1

129.6,1.4,1 4

5.5,1.9,-3.4 2

176

20.5,1.3,-15.8

1028.2,0.8,-19.0

118.2,-5.7,-11.5

1314,3.4,-4.4

20.2,5.1,-3.3

2118.4,9.4,-6.8 2

-1.0,6.9,-3.9 20.4,6.0,-4.4

2-0.6,6.2,-3.6 12

1.5,4.3,6.2

1919.8,6.5,-7.4

27.9,-0.2,-5.7 1

22.1,-6.6,-14.5

4

14.1,-7.4,-2.7

9

518.6,-6.6,-7.0

122.2,-6.7,-14.5

121.9,-6.5,-14.4

3925.6,8.5,4.9

119.9,-5.4,-13.5

1721.3,2.8,-7.0 19

28.0,-0.1,-8.9 14

15.4,-0.2,-11.2

18.4,-2.6,-4.8

820.7,-9.9,-2.8 10

30

8.8,-2.8,-5

9.4,-2.2,-5.2

32.1,1.2,-16.3

22.5,-8.6,-4.9

8.7,-3.3,-4.4

16.8,-4.1,-2.7

12,10.9,7.1



grade with respect to Melitaeini. The sister group ofNymphalinae (without Historis and Baeotus) is Bib-lidinae in the most parsimonious trees from the com-bined data set, although this clade has weak support(Fig. 1).

Results of the PBS analysis show that all three generegions are congruent at 52 of the 183 resolved nodes(Figs 1–4). These nodes generally have high BS values(range 3–63, mean 19) and tend to be concentrated inthe clades describing Nymphalini and Melitaeini. At

Figure 2. Relationships of sampled species from the tribe Nymphalini as delimited in Fig. 1. Tree statistics and branchthickness as in Fig. 1.

Aglais ioAglais milbertiAglais urticae

Antanartia abyssinica

Antanartia deliusAntanartia schaenia

Araschnia levana

Colobura dirce

Hypanartia bella

Hypanartia charonHypanartia kefersteini

Hypanartia letheHypanartia lindigii

Polygonia canace

Mynes geoffroyi

Nymphalis antiopaNymphalis californica

Nymphalis l albumNymphalis polychloros

Nymphalis xanthomelas

Polygonia c-album

Polygonia c-aureum

Polygonia comma

Polygonia egea

Polygonia faunus

Polygonia haroldi

Polygonia interrogationis

Polygonia oreas

Polygonia progne

Polygonia satyrus

Polygonia gracilis

Smyrna blomfildia

Symbrenthia lileaSymbrenthia hypatia

Symbrenthia hypselis

Tigridia acesta

Vanessa annabella

Vanessa atalanta

Vanessa braziliensis

Vanessa cardui

Vanessa gonerilla

Vanessa indica

Vanessa itea

Vanessa kershawi

Vanessa myrinna

Vanessa virginiensis

922.5,-8.6,-4.9

611.5,4.4,-9.9

1112.7,7.2,-8.9

421.5,-1.5,-16.0

419,-1.4,-13.5

821.2,-0.3,-12.9

165,0.9,10.1

269,11.4,

5.6

8

2,3.9,2.1

11,-1,1 6

4.5,-0.6,2.1 53.2,1.4,0.4 5

-4.5,5.4,4.1

23,-2.1,1.1

60.5,4.5,1.1

10.4,0.6,

011

5.6,4.2,1.2

79.3,-2.3,0 4

9.9,-5.2,-0.7 78,-1.1,0.1 1

7.3,-4.3,-2.0 17.2,-4.3,-1.9 4

5.4,-1.2,-0.2

3

2,1.9,-0.9

82.9,4.3,0.8

1-2.8,3.7,0.1

1-2.8,3.7,

0.17

2.9,4.6,-0.5

249.2,10.2,4.6 5

6, 2.1,1.1- 90.2,5.4,3.4

77.1,-1.1,

1.1

2511,6.9,

7.1

227,9.9,5.1

22,0.9,-0.9

1024.7,-4.4,

-10.3

6237,13.4,11.6

1723.9, 0.6,

-6.3- 2121.8,5.3,-6.2

93.6,2.7,

2.7

112.5,3.5,5.1 15

2.4,6.6,6.1

1933.6,-5.7,

-8.9



Figure 3. Relationships of sampled species from the tribes Junoniini and Victorinini, as delimited in Fig. 1, as well asspecies in the genera Kallimoides, Vanessula and Rhinopalpa. Tree statistics and branch thickness as in Fig. 1.

Anartia amatheaAnartia fatima

Anartia jatrophae

Hypolimnas alimena

Hypolimnas anthedon

Hypolimnas bolina

Hypolimnas misippus

Hypolimnas pandarus

Hypolimnas usambara

Junonia artaxia

Junonia coenia

Junonia erigoneJunonia iphita

Junonia natalica

Junonia oenoneJunonia sophia

Junonia terea

Junonia touhilimasa

Kallimoides rumia

Kamilla cymodoce

Metamorpha elissa

Napeocles jucunda

Precis andremiajaPrecis antilope

Precis archesiaPrecis ceryne

Precis cuama

Precis octaviaPrecis sinuata

Precis tugela

Protogoniomorpha anacardii

Protogoniomorpha cytora

Protogoniomorpha parhassus

Rhinopalpa polynice

Salamis antevaSalamis cacta

Siproeta epaphus

Siproeta stelenes

Vanessula milca

Yoma algina

1

22.2,-6.7,-14.5

319.9,-6.6,

-10.48

31.4,-6.9,-16.5

116.7,-5.6,-10.1 40

43.8,-0.7,-3.2 3340.6,0.1,-7.7

1627.5,-1.2,

-10.31

23.3,-8.0,-14.3

121.8,-6.4,-14.4

1927.2,1.1,-9.3

77.5,-0.3,

-0.2

179.7,-0.6,

7.9 115.5,4.3,1.2 2

2,-0.1,0.1 11,-0.1,0.1

726.8,-4.9,

-14.9

2531.3,1.7,-7.9

123.3,-7.4,-14.9

21.1,-0.2,1.1

23.2,-2.4,1.2 3

4,-1.1,0.1

23.0,-2.1,

1.1

423.6,-4.7,

-14.9

521.3,-5.6,-10.7

1314.5,-1.9,0.4

121.9,-6.5,-14.4

36.0,-2.1,

-0.916

29.2,0.3,-13.5

122,-6.5,-14.5

122.1,-6.6,-14.5

2335.2,-0.5,-11.7 4

20.6,-5.6,-11

2833.4,-1.5,

-3.9

2620,3.9,

2.1

3

6.9,0.5,-4.4 5

4.2,1.6,-0.8 3434,9.9,-9.9

6246.6,9.9,

5.5

418.4,-5.3,

-9.1



Figure 4. Relationships of sampled species from the tribe Melitaeini and Kallimini as delimited in Fig. 1. Tree statisticsand branch thickness as in Fig. 1.

Anthanassa ardysAnthanassa drusilla

Anthanassa otanes

Anthanassa texanaAnthanassa tulcis

Castilia eranitesCastilia ofella

Catacroptera cloanthe

Chlosyne acastus

Chlosyne cyneas

Chlosyne gaudealis

Chlosyne gorgone

Chlosyne harrisii

Chlosyne janais

Chlosyne lacinia

Chlosyne narva

Chlosyne nycteis

Chlosyne palla

Chlosyne theona

Doleschallia bisaltide

Eresia clio

Eresia coela

Eresia eunice

Eresia letitia

Eresia peloniaEresia quintilla

Eresia sestia

Euphydryas aurinia

Euphydryas chalcedona

Euphydryas desfontainii

Euphydryas editha

Euphydryas gillettiiEuphydryas phaeton

Gnathotriche exclamationisHigginsius fasciata

Janatella leucodesma

Kallima paralektaKallima inachus

Mallika jacksoni

Mazia amazonica

Melitaea arduinna

Melitaea britomartis

Melitaea cinxia

Melitaea deione

Melitaea didymoidesMelitaea latonigena

Melitaea persea

Melitaea scotosiaMelitaea punicaMelitaea trivia

Melitaea varia

Dymasia dymasTexola elada

Microtia elva

Phyciodes batesii

Phyciodes cocytaPhyciodes graphica

Phyciodes mylittaPhyciodes orseis

Phyciodes pallescens

Phyciodes pallida

Phyciodes phaon

Phyciodes picta

Phyciodes pulchella

Phyciodes tharos

Poladryas arachne

Tegosa anietaTegosa tissoides

Telenassa trimaculata

121.9,-6.5,

-14.4

3925.6,8.5,4.9

2221.1,8.8,-7.9

69.3,2.1,-5.4

5-1,5.9,0.1

3312.9,15.5,4.6

815.2,-0.9,-6.3

28.3,-0.4,-6.0

67.7,1.4,-3.1

30.9,1.1,

1 93,3.9,2.1 3

1.2,-0.4,2.2

41.7,1.2,

1.1

34.3,-0.3,-1

32,-0.1,

1.115

4.9,8.6,1.5

55.8,0.6,-1.4

52.5,2.5,0 1

3.1,-1.3,-0.8 1012,-1.7,-0.3

3023.2,3.7,

3.2

156.9,5,3.1

42.7,2.2,

-0.9

155.4,11.7,

-2.128

14.4,11.2,2.4

36,-2.1,-0.9 6

3,2.9,0.1

2-2.9,7,-2.1

2-1.3,7.9,-4.6 2

1,4.5,-3.5 117.4,5.4,-1.8 4

4,0.9,-0.9

187.9,10.7,

-0.5

5531.5,17.7,

5.8

163.4,10,2.6

123.7,9.1,

-0.8 2-1.3,2.5,0.8

11.9,-1,0.1

2-1.3,2.3,1 25

6,13.9,5.1 1310,0.9,2.1

136,5.9,1.1

2211,8.9,2.1 9

7,1.9,0.1

187.6,7.4,

3

3-0.3,4.2,

-0.9 1-2.1,3.9,-0.8

179.1,8.2,-0.3

610.3,0.1,-4.3 1

-1.5,3.5,-1 66.2,-0.3,0.1 1

-1,2.9,-0.9 86.6,1.8,-0.4 4

0,4.9,-0.9

88.7,0.5,

-1.2

2910.1,8.6,

10.422

7.1,14.3,0.6

163,7,6 4

6.9,1.9,-4.8

3624.7,19.7,

-8.4 97.3,5.1,-3.3 30

20.2,6.7,3.1

2830.6,3.8,

-6.52

3,-1.1,0.1

119.9,-5.4,

-13.5 1615.7,2.2,-1.9 17

29,-0.6,-11.4

3737.8,4.3,

-5.5



131 nodes, one or two gene regions provide signal thatis incongruent with the weight of the combined evi-dence. The different patterns of incongruence are sum-marized in Table 2. Nodes which show incongruencetend to have lower BS values (range 1–19, mean 7.7).Most of the incongruent nodes lie in the basalbranches of the cladogram (Fig. 1) and in the kal-limine grade (Figs 3, 4).

The PBS results show that the data partitions arecontributing unequally to the phylogenetic patternsinferred in this study. The COI data set conflicts at 17nodes, of which one node has a PBS score of <-4. Thetotal PBS for the COI data partition is 2176.1 (mean11.9). The EF1-a data partition is incongruent at 70 ofthe 183 resolved nodes, 23 of which have a PBS scoreof <-4, and has a total PBS of 370.3 (mean 2.0). Thewingless data alone have a negative PBS score at 106of the 183 resolved nodes, 64 of which have a PBSscore of <-4, and the gene region has a total PBS of -533.5 (mean -2.9).

Many of the clades are very stable (sensu Giribet,2003) and are present in a strict consensus of all treesfound across the different weighting schemes from theequally weighted analysis to the analysis with 10 : 1TV/TI weighting (Figs 1–4). These include the largernymphaline clade (including Biblidinae, Cyrestinae,Apaturinae and Nymphalinae; see Wahlberg et al.,2003b), Apaturinae and Biblidinae, Pseudergolini,Cyrestini, Nymphalini and Melitaeini (Fig. 1), andmany of the well-defined genera (Figs 2–4). Weightingtransversions 2 times transitions is the only analysiswhich gives a monophyletic Nymphalinae, with His-toris + Baeotus coming out as sister to the rest ofNymphalinae.

PHYLOGENETIC PATTERNS IN COEINI

The tribe Coeini does not form a monophyletic groupin any of the analyses (Figs 1, 2). In the combinedanalysis, Smyrna is sister to Nymphalini with good

support, Colobura and Tigridia are sister genera withstrong support and they are sister to Nymphalini +Smyrna. The monophyly of Smyrna, Tigridia,Colobura and Nymphalini has good support and is sta-ble, though there is conflict from the nuclear genes(Fig. 1). Historis and Baeotus form a monophyleticgroup with strong support and both genera are mono-phyletic (Fig. 1). Historis odius and H. acheronta (thelatter sometimes placed in the genus Coea, from whichthe tribal name is derived) are sister species with onlyweak support and moderate conflict from the twonuclear genes. Baeotus, on the other hand, is astrongly supported monophyletic group with no con-flict from the different data partitions. Constrainingthe five coeine genera to form a monophyletic groupresults in trees that are 19 steps longer than the mostparsimonious trees found for the combined data sets.Constraining Colobura, Tigridia and Smyrna to forma monophyletic group results in trees seven stepslonger than the most parsimonious trees.

The association of Colobura, Tigridia and Smyrnawith Nymphalini is stable under all the differentweighting schemes, as is the clade containing Historisand Baeotus species. However, the position of Historis+ Baeotus changes as one increases the weight oftransversions to transitions. When transversions areweighted 2 times transitions, this clade is sister toNymphalinae. At higher tested weighting schemes(transversions weighted 3–10 times transitions), His-toris + Baeotus appears as the sister clade to Apaturi-nae.

PHYLOGENETIC PATTERNS IN NYMPHALINI

Within the monophyletic Nymphalini, relationshipswithin the Nymphalis-group of genera (i.e. Nymphalis,Polygonia and Aglais) are almost identical to thosehypothesized in a previous study (Wahlberg & Nylin,2003). The exception is Polygonia canace, which is sis-ter to Nymphalis in the present study, whereas it wassister to the rest of Polygonia in the 2003 study. Wahl-berg & Nylin (2003) was based on an additional genesequence and a morphological data set and thus weprefer the hypothesis supported in that study. Polygo-nia oreas and P. haroldi were not included in previousphylogenetic studies of the Nymphalis-group, and ourresults suggest that the former is related to P. gracilisand the latter is sister to a clade containing P. oreas,P. gracilis and P. satyrus.

Other genera in Nymphalini are here sampled morebroadly than in previous studies and we are now ableto identify three additional clades within Nymphalini.Two species of Antanartia (A. delius and A. schaenia)form the sister group to the rest of Nymphalini. Thenext most basal clade is formed by the genera Ara-schnia, Mynes and Symbrenthia, which group together

Table 2. Number of nodes in the ingroup at which differ-ent patterns of incongruence were observed. +, PBS scorepositive or 0; -, PBS score negative (magnitude not takeninto account)

Number of nodes COI EF1-a wingless

47 + + +46 + + –20 + – +

6 – + +56 + – –

0 – – +8 – + –



with good support. The third major new clade com-prises the genera Vanessa and Hypanartia, thoughthis clade has only moderate support and is not stable.Vanessa + Hypanartia is sister to the Nymphalis-group clade. A surprising result is the position ofAntanartia abyssinica within Vanessa; it is not at allclosely related to the other Antanartia species. WithinVanessa, species often placed in the genus Cynthia donot form a monophyletic group, as one species,V. annabella (and presumably its putative sister spe-cies V. carye; Shapiro & Geiger, 1989), appears to besister to the rest of Vanessa.

Sensitivity analyses suggest that the relationshipswithin the Nymphalis-group are very stable; the onlynode which is unstable is the sister relationshipbetween Polygonia egea and P. c-album + P. faunus.Hypanartia is stable, as are the relationships of spe-cies within the genus. Within Vanessa, the sister rela-tionship of A. abyssinica to V. atalanta + V. indica isstable, as is the sister species relationship ofV. gonerilla + V. itea, and the V. cardui clade (V. carduito V. myrinna in Fig. 2). Other stable clades areAntanartia delius + A. schaenia and Symbrenthia.

PHYLOGENETIC PATTERNS IN KALLIMINI

The tribe Kallimini as currently circumscribed doesnot form a monophyletic group, but rather a paraphyl-etic grade with regard to Melitaeini. Constraining it tobe monophyletic results in trees that are four stepslonger than the most parsimonious trees found for thecombined data set. The strict consensus of the mostparsimonious trees shows two clades. The basal cladecontains the following subclades: (1) a largely Neotro-pical subclade including Anartia, Siproeta, Napeoclesand Metamorpha (termed the Anartia-clade), (2) theAfrican Kallimoides, a subclade with Vanessula andRhinopalpa (both monotypic genera that have not pre-viously been associated with each other or indeed withany other kallimine genera) and (3) a subclade thatcontains the largely African and Asian generaJunonia, Kamilla, Precis, Hypolimnas, Salamis, Pro-togoniomorpha and Yoma (termed the Junonia-clade).The second clade in the Kallimini grade, which is sis-ter to the tribe Melitaeini, includes the African Catac-roptera and Mallika, the Asian Kallima and theAustralasian Doleschallia (termed the Kallima-clade).

Weighting transversions more than transitionscauses Kallimoides and Vanessula to become sistertaxa. These two form the sister clade to Rhinopalpaand the Anartia- and Junonia-clades when transver-sions are weighted 2 or 3 times transitions. However,weighting schemes of 5 and above cause Kalliomoides,Vanessula, Rhinopalpa and the Anartia-clade tobecome the most basal clades in the entire tree, afterthe outgroups Heliconius and Adelpha.

Within the Anartia-clade, Napeocles and Siproetaform a strongly supported, stable subclade, as do thethree species of Anartia. The sister group of Anartiamay be either Siproeta/Napeocles or Siproeta/Napeo-cles + Metamorpha. It is clear that Metamorpha is anentity distinct from Siproeta (as suggested by Fox &Forbes, 1971), though historically Siproeta steleneshas occasionally been placed in Metamorpha. Theclade containing these neotropical genera is stable insensitivity analyses. The position of Kallimoides asthe most basal taxon of this clade has only weak sup-port and the node is not stable, suggesting that itsplacement requires further investigation.

The Junonia-clade exhibits some of the greatest sur-prises of this study. The first is that Junonia and Pre-cis are separate genera that are not even sistergroups. The genus Kamilla is clearly within Junonia,as was concluded by Shirôzu & Nakanashi (1984)based on morphology (but see Larsen, 1991). In addi-tion, the species Protogoniomorpha cytora is foundwithin Junonia, rather than with other speices of Pro-togoniomorpha or Salamis, with which it has alwaysbeen associated. The sister group relation betweenPrecis and Hypolimnas has good support and is stable,except when transversions are weighted 3 times tran-sitions, when it is sister to the Salamis + Yoma + Pro-togoniomorpha + Junonia clade. Another surprise isthat the African Protogoniomorpha, which has beenusually considered to be a synonym of Salamis, is thesister group to the Australasian Yoma, and that Sala-mis is the sister group to Protogoniomorpha + Yoma.Vári (1979) has argued that species of Protogoniomor-pha should not be considered to be congeneric withspecies of Salamis based on genitalic differences, aposition corroborated by our data. The sister grouprelationship of Yoma and Protogoniomorpha has goodsupport and is stable. To complete the surprise, Sala-mis, Yoma and Protogoniomorpha, usually consideredto be related to Hypolimnas, are placed in our resultsas sister group to Junonia, though this clade disap-pears when transversions are weighted 5 or moretimes transitions.

The implied position of the Kallima-clade as sisterto the Melitaeini is somewhat surprising given tradi-tional classifications, but this grouping has weak sup-port and may be due to long branch attraction. Indeedthe sister group relationship is not stable and disap-pears when transversions are weighted 2 or moretimes transitions. The relationships between Catac-roptera, Mallika and Kallima are strongly supportedand stable. The strongly supported, stable sister rela-tionship of Catacroptera and Mallika (both monotypic)has been suggested earlier by Shirôzu & Nakanishi(1984). Catacroptera and Mallika are usually associ-ated with Junonia rather than Kallima (Larsen,1991). The recently suggested (Parsons, 1999) associ-



ation of Doleschallia with Kallima is corroborated byour data, although support is not strong and there isstrong conflict between the nuclear genes and themitochondrial gene regions.

PHYLOGENETIC PATTERNS IN MELITAEINI

The monophyly of Melitaeini is very strongly sup-ported and stable, as are Euphydryas, Melitaea,Phyciodina and the sister relationship between Gna-thotriche and Higginsius (termed the Gnathotriche-group). The Chlosyne-group has moderate support butis unstable, with both the COI and wingless generegions conflicting with the EF1-a data. The genusChlosyne itself is a strongly supported, stable clade.The position of Euphydryas as the sister group to therest of the melitaeines is strongly supported and sta-ble, in agreement with previous studies (Kons, 2000;Wahlberg & Zimmermann, 2000). Relationshipsbetween the Chlosyne-, Melitaea-, Gnathotriche-groups and Phyciodina are not well-supported and areunstable. The most parsimonious trees from theequally weighted analysis suggests that the Gnathot-riche-group is sister to Phyciodina with moderate sup-port, and that the Melitaea-group is sister to these twowith good support (Fig. 4). This arrangement is stablewhen transversions are weighted 2 and 3 times tran-sitions, but breaks down at higher weights. The sisterrelationship of the Gnathotriche-group and Phyciod-ina is not stable and disappears after weights of 3times. This study presents a third novel arrangementof these four clades, with Kons (2000) having the Chlo-syne-group sister to Phyciodina + Gnathotriche-groupand Wahlberg & Zimmermann (2000) having theChlosyne-group sister to the Melitaea-group.

The positions of two genera (Poladryas and Higgin-sius) are not in agreement with previous studies. Incontrast to Wahlberg & Zimmermann (2000) but inagreement with Kons (2000), Poladryas is here relatedto the Chlosyne-group. Morphological evidence thatPoladryas is associated with Higginsius (Kons, 2000)is quite decisively contradicted by molecular evidence,which places Higginsius as sister to Gnathotriche withstrong support and stability. Within the Chlosyne-group, the genera Microtia, Texola and Dymasia forma strongly supported, stable monophyletic group, asfound by Kons (2000).

Relationships of the Neotropical Phyciodina (repre-sented by the genera Mazia, Tegosa, Eresia, Castilia,Telenassa, Janatella and Anthanassa) are generallynot well-supported or stable at the deeper nodes. Ere-sia, Castilia, Telenassa, Janatella and Anthanassa doform a stable monophyletic group to the exclusion ofTegosa and Mazia, even though the wingless data par-titions is in conflict with the COI and EF1-a data atthe node.

DISCUSSION

This is only the second comprehensive phylogeneticanalysis of the relationships within a nymphalidsubfamily, and the first to use molecular data. Thesole previous attempt to infer relationships within anymphalid subfamily is the recently published studyby Penz & Peggie (2003) on Heliconiinae. This is notaltogether surprising, given that the circumscrip-tions of the subfamilies have only recently stabi-lized (Harvey, 1991; Wahlberg et al., 2003b), andbecause the high degree of variation in morphologi-cal characters among species in Nymphalidae hasconfounded previous attempts to delineate naturalgroups (de Jong et al., 1996). In our study, we havebeen able to identify clades that are well-supportedby the three gene regions and stable to varied char-acter state transformation weights, as well as cladesthat are less robust and therefore likely to changewith the addition of more data. Based on ourresults, we are proposing a new classification of thesubfamily Nymphalinae, which is shown in Appen-dix 2.

THE CIRCUMSCRIPTION OF NYMPHALINAE

We have found that Nymphalinae as currentlycircumscribed is not monophyletic, although Nym-phalini, Kallimini and Melitaeini do form a mono-phyletic group with the inclusion of three generatraditionally placed in Coeini (i.e. Colobura,Tigridia and Smyrna). Two genera also tradition-ally placed in Coeini (Historis and Baeotus) areclearly not related to these three genera and indeeddo not appear to be closely related to Nymphali-nae. Our results are not unprecedented, as Muys-hondt & Muyshondt (1979) argued that based onlarval morphology Smyrna should be placed inNymphalini and that Colobura is closer to Nympha-lini than Historis and Baeotus. However, the sug-gestion by Muyshondt & Muyshondt (1979) thatCoeini is an unnatural group has not been followedby subsequent authors. On the other hand, Freitas& Brown (2004) found that, based on morphologi-cal data, Historis, Colobura and Smyrna formed amonophyletic group sister to Hypanartia and Van-essa. We were unable to sample the remainingostensibly coeine genus, Pycina, which has morpho-logical features in larvae and pupae that appear tobe intermediate between those in Historis/Baeotusand Colobura (Muyshondt & Muyshondt, 1979). Theunstable behaviour of the Historis + Baeotus cladein our study and its quite different position in thestudy by Freitas & Brown (2004) suggests thatmore data are needed to clarify its relationshipwithin Nymphalidae.



THE CIRCUMSCRIPTION OF NYMPHALINI

The well-supported association of Colobura, Tigridiaand Smyrna with Nymphalini suggests that theyshould be incorporated into the tribe, though the tra-ditional delineation of the tribe is also well-supportedand, in addition, is stable.

The placement of Antanartia abyssinica within Van-essa is clear. Vanessa (including A. abyssinica) is awell-supported clade and the sister relationship ofA. abyssinica to V. atalanta + V. indica is stable.Indeed, larval morphology of A. abyssinica corrobo-rates our evidence that it is unrelated to otherAntanartia species and related to Vanessa (Nakanishi,1989). Antanartia has been divided into two species-groups, the delius-group, comprising A. delius,A. schaenia and the unsampled A. borbonica, and thehippomene-group, to which A. abyssinica, and theunsampled A. hippomene and A. dimorphica belong(Howarth, 1966). Whether A. hippomene andA. dimorphica should also be placed in Vanessa is notclear; Nakanishi (1989) noted that the larvae ofA. abyssinica differed greatly from those ofA. schaenia and A. hippomene. Clearly, the missingspecies need to be sampled.

The divergent position of Antanartia with respect toHypanartia is also surprising, as all species wereincluded in Hypanartia prior to the description ofAntanartia by Rothschild & Jordan (1903). Antanartiahas always been assumed to be the sister group ofHypanartia (e.g. Willmott et al., 2001). However, ourresults suggest that the sister group to Hypanartia isVanessa and that Antanartia is sister to all the rest ofthe traditionally recognized Nymphalini. These posi-tions are stable when transversions are weighted 2and 3 times transitions, but they break down at higherweights. Weighting 5 and 7 times suggests thatAntanartia is sister to the Symbrenthia/Mynes/Ara-schnia clade and that Hypanartia is sister to these. Inno analysis does Antanartia appear as the sister toHypanartia.

Vanessa is usually thought to be the sister group tothe Nymphalis-group of genera (Wahlberg & Nylin,2003). Our study suggests that Vanessa is sister toHypanartia and that these two clades make up the sis-ter group to the Nymphalis-group. This arrangementis stable when transversions are weighted 2 and 3times transitions, while at higher weights only Van-essa is sister to the Nymphalis-group. Thus, it is likelythat either Vanessa or Vanessa + Hypanartia is morerelated to the Nymphalis-group than to the other gen-era in Nymphalini, and further new characters(molecular and morphological) will help in refining therelationships further.

The relationships of Symbrenthia, Mynes and Ara-schnia are rather surprising, especially since the sis-

ter relationship between Mynes and Araschnia isstable up to a weighting of transversions 7 times tran-sitions. A recent morphological study of this groupfound that Mynes is within Symbrenthia (Fric et al.,2004). Normally, Symbrenthia has been associatedwith Mynes (Parsons, 1999; Nylin et al., 2001; Wahl-berg & Nylin, 2003). For our results to be congruentwith those of Fric et al. (2004), we would have to findthat Mynes geoffroyi is sister to Symbrenthia hypselis,which is clearly not the case. Obviously, more speciesof all three genera need to be sampled to resolve theconflicting results of the two studies.

This study contains a much more comprehensivesampling of species in Nymphalini than our two pre-vious studies (Nylin et al., 2001; Wahlberg & Nylin,2003). The Nymphalis-group continues to be a well-supported and stable clade, though the stable positionof Polygonia canace as sister to species in the genusNymphalis is in contrast to earlier results. Polygoniacanace is usually placed in its own genus Kaniska, andthe uncertainty of its position may warrant the use ofthat genus name. However, our study confirms the sta-ble relationship of Aglais io (usually placed in its owngenus Inachis) with other species of Aglais and thestable relationship of Nymphalis l-album with otherspecies of Nymphalis. Also stable and well-supportedis the sister relationship of Polygonia and Nymphalis,with Aglais as sister to these two. This study includestwo species of Polygonia that have not been includedin previous phylogenetic studies, P. oreas andP. haroldi. The position of P. oreas as sister toP. gracilis is surprising as it is often considered to be asubspecies of P. progne (Scott, 1984). Polygoniaharoldi is a little known species and morphologically itis somewhat intermediate between P. progne andP. satyrus. The relationships of species in Polygoniaare being investigated in more detail currently (E.Weingartner, N. Wahlberg & S. Nylin, unpubl. data).

THE FATE OF KALLIMINI

The monophyly of Kallimini appears to be doubtful.The tribe, as previously circumscribed, never formed amonophyletic group in our analyses. Species belongingto Kallimini have occasionally been placed in highertaxa of their own, and our results show that some ofthese groups are strongly supported and stable. Thesewell supported groups of genera should be recognizedat the tribal level, unless one cares to resort to the (inour opinion, unacceptable) synonymization of Kal-limini with Melitaeini. Names are available for twotribes: Victorinini Scudder, 1893 for the Anartia-cladeand Junoniini Reuter, 1896 for the Junonia-clade. TheKallima-clade constitutes the newly circumscribedtribe Kallimini. The three remaining genera (Kalli-moides, Vanessula and Rhinopalpa) have to remain



incertae sedis until their positions are stabilized byfurther investigation.

There are 11 species in four genera in the newly cir-cumscribed tribe Victorinini. The close relationship ofNapeocles and Siproeta has never been considered,though Metamorpha has been considered to be aderived Siproeta (Fox & Forbes, 1971). Perhaps thesuperficial similarity of Metamorpha to Siproeta andthe highly distinctive wing patterns of Napeocles hasled previous investigators to ignore the latter. How-ever, weighting transversions higher than transitionscauses Napeocles to become sister to S. stelenes, whichis also recovered in some of the most parsimonioustrees of the equally weighted analysis. All weightingschemes other than equal weighting also recover a sis-ter relationship between Metamorpha and Anartia,suggesting that Metamorpha is not as closely relatedto Siproeta as previously thought. The relationships ofthe three species of Anartia included in this study arein concordance with a previous study on the genus(Blum et al., 2003). It is clear from our study that Vic-torinini is a well-defined entity that deserves the rankof tribe in the Nymphalinae.

The Junoniini, as circumscribed here, is also a well-defined and stable group. The relationships of the gen-era in Junoniini found in the equally weighted analy-sis are stable under fairly severe weighting schemes(transversions weighted up to 5 times transitions). Athigher weights, the Yoma + Protogoniomorpha cladebecomes sister to the Precis + Hypolimnas clade, andSalamis is sister to the rest of Junoniini. The closerelationships of Protogoniomorpha and Yoma and Pre-cis and Hypolimnas are stable under all weightingschemes. The clean separation of Precis and Junoniain our study is both surprising and gratifying. Thesetwo genera have long been conflated in the literature,despite their biogeographical disjunction and the factthat de Lesse (1952) showed clear genitalic differencesbetween them. de Lesse’s (1952) delimitations of thetwo genera are corroborated here. For North Americanresearchers it is important to emphasize that all NewWorld species (including the well-studied J. coenia)belong to Junonia and that Precis is restricted toAfrica.

The once common concept of Kallima (i.e. containingKallimoides, Kamilla and Mallika) is clearly untena-ble, as firmly concluded by Shirôzu & Nakanishi(1984). The Kallimini, as delimited here, contains onlyfour genera: Kallima, Catacroptera, Mallika and Dole-schallia.

THE DELINEATION OF MELITAEINI

The monophyly of Melitaeini is beyond doubt. Allthree gene regions support the clade and it is stable ina variety of weighting schemes. The sister group to

Melitaeini appears to be the newly circumscribed Kal-limini and/or Junoniini. Within Melitaeini, the sisterrelationship of Euphydryas to the rest of Melitaeini isin agreement with all previous studies and has a clear,well-supported, stable position in this study. The restof Melitaeini appears to be divided up into four dis-tinct groups: Phyciodina, Melitaeina (including onlyMelitaea), the Gnathotriche-group and the Chlosyne-group. Of these, only the Chlosyne-group is not stable,though Poladryas remains associated with Chlosynewith transversions weighted up to 7 times transitions.The Microtia + Texola + Dymasia clade becomes thesister to the rest of Melitaeini (excluding Euphydryas)when transversions are weighted 5 and 7 times tran-sitions, though it returns to being sister to Chlosyne atthe highest weighting scheme. The close relationshipof Microtia, Texola and Dymasia was also found byKons (2000), who suggested that Texola and Dymasiashould be synonymized with Microtia.

The close relationship of Higginsius to Gnathotrichewas suggested by Higgins (1981), but Kons (2000)found Higginsius to be associated with Poladryas. Ourresults strongly corroborate Higgins’ (1981) hypothe-sis. The two genera are very curious members of theAndean fauna and appear to be always rare (Higgins,1981). They comprise a total of six species and, withthe unsampled Caribbean Atlantea and Antillea (alsovery species-poor and always rare), create somethingof a biogeographical mystery. We find that the sisterrelationship of Gnathotriche and Higginsius to Phycio-dina is stable with transversions weighted up to 7times transitions. When transversions are weighted10 times transitions, the Gnathotriche-group becomessister to Melitaea. Since Phyciodina is largely a Neo-tropical subtribe, its close relationship to the Gnathot-riche-group is perhaps not surprising, but the possibleorigin of both groups in South America requires fur-ther investigation.

Of the six missing genera, four (Tisona, Dagon, Orti-lia and Phystis) putatively belong to the subtribe Phy-ciodina (Higgins, 1981). The remaining two genera,Antillea and Atlantea, are restricted to the GreaterAntilles in the Caribbean and have until recently notbeen associated with other melitaeine genera. Kons(2000) placed both in the Chlosyne-group, withAtlantea being sister to Higginsius. It is clear thatthese two genera need to be sampled for moleculardata.

A NOTE ON PHYLOGENETIC PATTERNS IN THE OUTGROUPS

Although our study has concentrated on Nymphali-nae, we have extensively sampled several potentialsister groups to the subfamily. In particular, we havesampled all genera in the tribes Cyrestini and Pseud-

240

N. WAHLBERG

ET AL

.



2005,

86

, 227–251

ergolini, which constitute the Cyrestinae in Wahlberg

et al

. (2003b). However, in the present study the twotribes do not form a monophyletic group; rather,Cyrestini appears as the most basal group in thenymphaline clade and Pseudergolini is sister to Apa-turinae (Fig. 1). There are four clades in the outgroupappearing with good to strong support that correspondto the Apaturinae, Biblidinae, Cyrestini and Pseuder-golini. However, the relationships among these fourtaxa and Nymphalinae are very poorly supported, areunstable and show much conflict among partitions.Clearly, the nymphaline clade (

sensu

Wahlberg

et al

.,2003b) needs to be more extensively sampled andmore data need to be added before any robustconclusions emerge about relationships of the majorclades.

B

IOGEOGRAPHICAL

HISTORY

OF

N

YMPHALINAE

Despite the limitations of our current phylogenetichypothesis, some strong patterns emerge from our dis-persal-vicariance analysis (Fig. 5). First of all, manydispersal events are required to explain the distribu-tion patterns seen today: 44 when the maximum num-ber of ancestral areas is not constrained and 46 whenthey are constrained to two. This should not be sur-prising, since Nymphalinae contains some of the mostmobile butterflies known, such as

Vanessa cardui

,which is famous for being able to disperse over thou-sands of kilometers in one generation. Indeed, thegenus

Vanessa

, which is present in all the major zoo-logical biomes of the world, has a highly ambiguousreconstruction of its ancestral distribution. Perhapsthe ancestor of

Vanessa

was a widespread species,much like

V. cardui

today, that subsequently speciatedduring intervals of isolation due to climatic or otherfactors.

The distribution of the most recent common ances-tor of Nymphalinae (excluding

Historis

and

Baeotus

)appears to be widespread (Fig. 5), though this may bean artefact of the program used and might be fixed byincluding outgroups with known ancestral distribu-tions (Ronquist, 1996). We refrained from doing so, asour sampling of the outgroups is not sufficient toresolve their ancestral distributions; also, we are notconfident about the current hypothesis of relation-ships among major groups in the nymphaline clade.

The tribe Nymphalini may have originated in SouthAmerica, one of the seven possible ancestral areas inthe unconstrained analysis (Fig. 5) and one of two inthe constrained searches. Africa may have been colo-nized from South America by the common ancestor of

Smyrna

and the rest of Nymphalini. Once the lineageleading to

Smyrna

split off, it appears that the Palae-arctic was colonized from Africa by the common ances-tor of

Antanartia

and the rest of Nymphalini

(excluding

Colobura, Tigridia

and

Smyrna

). An alter-native scenario would have a widespread commonancestor in South America, Africa and the Palaearcticthat then speciated through a series of vicarianceevents (Fig. 5). The Palaearctic has, however, been avery important area for the diversification of the gen-era

Aglais, Nymphalis, Polygonia

and possibly

Van-essa

. There have clearly been several independentcolonizations of the Nearctic from the Palaearctic byspecies in the the former three genera.

The clade that includes the tribes Junoniini and Vic-torinini, as well as

Kallimoides, Rhinopalpa

and

Van-essula

, appears to have originated in Africa (Fig. 5).There is a colonization of South America from Africaby the ancestor of

Kallimoides

and Victorinini. Also,the Oriental region appears to have been colonizedindependently several times. The patterns in thisclade are obscured somewhat by the weak support forthe current hypothesis of relationships; in addition,many missing species in the genera

Hypolimnas

and

Junonia

are likely to have had a decisive effect on theresults due to their presence in the African, Orientaland Australasian areas. However, it is clear that theAfrican region has been instrumental in the diversifi-cation of taxa in the clade.

The tribe Kallimini appears to have originated inthe Oriental region, with a colonization of Africa bythe common ancestor of

Kallima

and

Catacroptera/Mallika

(Fig. 5). This hypothesis needs to be tested byincluding the several species of

Doleschallia

foundexclusively in the Australasian region.

The Nearctic region has been an important area forthe diversification of the tribe Melitaeini (Fig. 5). Oneinterpretation of the patterns recovered by DIVA isthat the group originated in the Nearctic and subse-quently colonized the Palaearctic (the ancestor of

Melitaea

) and the Neotropics (ancestor of the

Gnathot-riche

-group and Phyciodina). Interestingly, the cur-rent phylogenetic hypothesis suggests a disjunctdistribution for the ancestor of

Melitaea

+

Gnathot-riche/Higginsius

+

Phyciodina being found in thePalaearctic and the Neotropics. Sampling the twomissing Caribbean genera will be important to under-standing the historical biogeography of Melitaeini.

In summary, it is clear that several regions havebeen important areas of diversification of clades inNymphalinae, viz. the Palaearctic for the

Nymphalis

-group of genera, Africa for Junoniini and related gen-era and the Nearctic for Melitaeini. Our DIVA analy-sis indicates that dispersal has had a major effect onthe distributions of extant species in Nymphalinae.Whether dispersal or vicariance has been the moreimportant process in generating the deeper diver-gences in Nymphalinae can be tested by dating diver-gences using molecular clocks calibrated with fossils.Such a study is in preparation and the implications of



Figure 5. Reconstructed ancestral distributions according to a dispersal-vicariance analysis. Letters after the names oftaxa give the current distribution of those taxa.

Colobura/Tigridia BSmyrna B

Antanartia C

Araschnia DMynes F

Symbrenthia E

Hypanartia BVanessa annabella A

Vanessa abyssinica CVanessa atalanta ADVanessa indica DE

Vanessa gonerilla/itea F

Vanessa cardui ACDEVanessa kershawi F

Vanessa virginiensis ABVanessa braziliensis/myrinna B

Aglais io DAglais milberti AAglais urticae D

Polygonia canace DENymphalis l-album AD

Nymphalis polychloros DNymphalis xanthomelas D

Nymphalis californica ANymphalis antiopa AD

Polygonia c-aureum D

Polygonia egea DPolygonia faunus APolygonia c-album D

Polygonia North American clade A

Kallimoides rumia CVictorinini B

Rhinopalpa EVanessula C

Hypolimnas anthedon/usambara CHypolimnas misippus CEF

Hypolimnas alimena FHypolimnas bolina CEFHypolimnas pandarus F

Precis C

Salamis CYoma EFProtogoniomorpha CJunonia erigone FJunonia iphita EJunonia artaxia/touhilimasa/cytora C

Junonia cymodoce CJunonia terea/natalica C

Junonia coenia AJunonia sophia/oenone C

Doleschallia EFCatacroptera/Mallika CKallima E

Euphydryas gilllettii AEuphydryas aurinia/desfontainii D

Euphydryas North American clade A

Poladryas AMicrotia elva ABTexola/Dymasia AChlosyne Central American clade AB

Chlosyne cyneas AChlosyne theona A

Chlosyne nycteis A

Chlosyne gorgone AChlosyne lacinia AB

Chlosyne acastus/palla A

Melitaea DGnathotriche/Higginsius B

Mazia BTegosa B

Phyciodes A

Anthanassa texana AAnthanassa tulcis B

Anthanassa rest BEresia/Castilia/Telenassa B

DFDEEF

DEF

DCD

B

DDF

CDF

ADACDADF

ACDF

ADD

ADD

DD

D

ADD

ADD

D

D

D

DDF

DEF

CDCDF

CDEF

BCBD

BCDBDF

BCDFBDEF

BCDEF

BBD

BCDBDF

BCDFBDEF

BCDEF

BC

CE

FF

FCF

CEF

CCF

C

CEC

EF

ACC

CC

CECF

CEF

C

C

C

C

CEE

ADA

A

AA

AA

AA

A

A

ABB

BAB

BB

BBD

ABAD

ABD

AAD

ABD

AEADE

ABDE

ACACE

ACDEABCDE

ABCEACDE

ABCDEACDEF

ABCDEF

A = NearcticB = NeotropicsC = AfrotropicsD = PalaearcticE = OrientalF = Australasia

AF CF ACF DFADF CDF ACDF EF

AEF CEF ACEF DEFADEF CDEF ACDEF

BD ABD BCD ABCDBDE ABDE BCDE

ABCDE BF ABF BCFABCF BDF ABDF BCDFABCDF BEF ABEF BCEF

ABCEF BDEF ABDEFBCDEF ABCDEF

F DF ADF BDF ABDF CDF ACDFBCDF ABCDF DEF ADEF BDEF ABDEF

CDEF ACDEF BCDEF ABCDEF

BDABDBCD

ABCDBDF

ABDFBCDF

ABCDF



it will be discussed elsewhere (N. Wahlberg, unpubl.data).

PBS AND SENSITIVITY ANALYSIS

The ease with which molecular data can be generatedhas led to an ever-increasing number of publishedphylogenetic studies. Common to almost all thesestudies is the lack of sensitivity analyses and, in thecase of multiple independent data sets, evaluation ofcongruence at given nodes (but see Reed & Sperling,1999). An exception to the former are studies based ondirect optimization of ribosomal DNA sequences(Wheeler, 1995; Giribet, 2003). Congruence amongdata partitions is usually evaluated at the level ofentire data sets through tests such as the ILD (Farriset al., 1994), rather than through evaluation at everyresolved node. Again, a few exceptions are notable (e.g.Gatesy et al., 1999; Cognato & Vogler, 2001; Lambkinet al., 2002; Damgaard & Cognato, 2003; Wahlberg &Nylin, 2003). Both the PBS and sensitivity analysisallow detailed evaluation of which nodes are likely tobe robust and stable to the addition of new data, aswell as which nodes require further investigation. Weemphasize that these are heuristic tools to assess thequality of the most parsimonious hypothesis and guidefuture sampling of characters and taxa, rather thanalternate analytical approaches that challenge thephilosophical basis of our preferred optimalitycriterion.

CONCLUSION

The main conclusions of this study are: (1) Smyrna,Colobura and Tigridia are associated with Nympha-lini, (2) ‘Kallimini’ is more closely related to Melitaeinithan to Nymphalini, (3) ‘Kallimini’ is not monophyl-etic, but is paraphyletic with regard to Melitaeini, (4)Melitaeini and Nymphalini plus the three coeine gen-era are strongly supported monophyletic groups, and(5) Precis and Junonia are not synonymous or evensister groups.

Major unanswered questions are: (1) the positions ofHistoris + Baeotus in the nymphaline clade, and (2)the sister group of Nymphalinae. Increased samplingof species in the subfamilies Biblidinae and Apaturi-nae, as well as increased character data (molecularand morphological), will help to resolve thesequestions.

ACKNOWLEDGEMENTS

As always, this study would not have been possiblewithout the truly wonderful, altruistic help of bothamateur and professional lepidopterists. We are eter-nally grateful to Jun-ichi Asahi, Alexei Belik, Anton

Chichvarkhin, Tim Davenport, Phil DeVries, AndréFreitas, Daniel Janzen, Chris Jiggins, Tjeerd Jonge-ling, Darrell Kemp, Norbert Kondla, Runar Krogen,Gerardo Lamas, Torben B. Larsen, Yi-Hsin Lee, JorgeLeón-Cortés, Mike Leski, Debra Murray, NaomiPierce, James Scott, Mike Soukop, Constant’ Stefa-nescu, Martin Steinbauer, Man-Wah Tan, BruceWalsh, Andy Warren, and Keith Willmott for helpingus to obtain specimens. We thank Tiina Berg and Sim-ina Vintila for expert help in the lab. We also thankAndy Warren, Keith Willmott, Carla Penz and TorbenB. Larsen for useful comments on a draft version of themanuscript.

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Willmott KR, Hall JPW, Lamas G. 2001. Systematics ofHypanartia (Lepidoptera: Nymphalidae: Nymphalinae),with a test for geographical speciation mechanisms in theAndes. Systematic Entomology 26: 369–399.

Zimmermann M, Wahlberg N, Descimon H. 2000. Phy-logeny of Euphydryas checkerspot butterflies (Lepi-doptera: Nymphalidae) based on mitochondrial DNAsequence data. Annals of the Entomological Society ofAmerica 93: 347–355.



AP

PE

ND

IX 1

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HE

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AE

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hec

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70-6

CO

ST

A R

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tter

fly

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AY

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LIM

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INA

EA

del

pha

bred

owi

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107-

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AY

7885

91A

Y78

8693

AY

7884

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TIN

AE

Cyr

esti

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100-

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AN

GL

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et D

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2182

40A

Y21

8260

AY

2182

78C

her

son

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ria

NW

111-

3IN

DO

NE

SIA

: Su

law

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Sek

oA

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8601

AY

7887

03A

Y78

8465

Mar

pesi

a or

silo

chu

sR

B25

0B

RA

ZIL

: Ron

don

ia, A

riqu

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AY

7886

04A

Y78

8706

AF

2465

32M

arpe

sia

chir

onR

B22

7B

RA

ZIL

: Ron

don

ia, A

riqu

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AY

7886

03A

Y78

8705

AY

7884

67A

mn

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dec

ora

NW

101-

1IN

DO

NE

SIA

: W. S

um

atra

, Bra

stag

iA

Y21

8235

AY

2182

54A

Y21

8273

Dic

hor

ragi

a n

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ach

us

NW

111-

10P

HIL

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INE

S: S

. Ley

te, H

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nan

gan

AY

7886

02A

Y78

8704

AY

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66P

seu

der

goli

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edah

NW

118-

1V

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NA

M: L

ao C

aiA

Y78

8605

AY

7887

07A

Y78

8468

Sti

boch

ion

a n

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NW

100-

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AN

GL

AD

ES

H: S

ylh

et D

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ion

AY

2182

49A

Y21

8269

AY

2182

87B

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IDIN

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Ari

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W82

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UG

AN

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For

est

AY

2182

37A

Y21

8256

AY

2182

74B

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raN

W88

-14

ZIM

BA

BW

E: M

aron

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AY

7885

95A

Y78

8697

AY

7884

60C

alli

core

pac

ifica

NW

119-

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OS

TA

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Y78

8596

AY

7886

98A

Y78

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Cat

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e n

um

ilia

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CO

ST

A R

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AY

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BR

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ondo

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Y78

8597

AY

7886

99A

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6581

Eu

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W82

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AN

ZA

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: Am

ani

AY

2182

42A

Y21

8262

AY

2182

80H

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NW

62-3

CO

ST

A R

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: Bu

tter

fly

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su

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Y09

0216

AY

0901

82A

Y09

0149

Mes

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tha

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hea

NW

83-5

UG

AN

DA

: Kib

ale

For

est

AY

7885

98A

Y78

8700

AY

7884

62M

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lia

cape

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W10

9-4

EC

UA

DO

RA

Y78

8599

AY

7887

01A

Y78

8463

Nic

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laN

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PE

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: Roa

d to

Yu

rim

agu

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Y21

8245

AY

2182

65A

Y21

8283

Pan

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NW

109-

8E

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AD

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AY

7886

00A

Y78

8702

AY

7884

64S

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BA

BW

E: H

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8247

AY

2182

67A

Y21

8285

AP

AT

UR

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EA

patu

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ris

NW

69-6

Bu

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Y09

0199

AY

0901

65A

Y09

0132

Ast

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ampa

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lia

NW

82-1

5U

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zon

aA

F18

7734

AY

2182

57A

Y21

8275

Eu

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MA

LA

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IA (

MC

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AY

7885

93A

Y78

8695

AF

2465

88M

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SIA

: Pri

mor

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Y78

8594

AY

7886

96A

Y78

8459

Tim

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a m

acu

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NW

97-8

TA

IWA

N: T

aitu

ng

Cou

nty

AY

2182

51A

Y21

8271

AY

2182

89C

hit

oria

ch

ryso

lora

NW

97-1

1T

AIW

AN

: Tai

tun

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oun

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Y78

8592

AY

7886

94A

Y78

8458

NY

MP

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LIN

AE

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EC

UA

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R: P

asta

za, L

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iA

Y78

8613

AY

7887

18A

Y78

8479

Bae

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s d

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lion

NW

130-

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CU

AD

OR

: Nap

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Y78

8616

AY

7887

21A

Y78

8482

Bae

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W13

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EC

UA

DO

R: S

ucu

mbi

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AY

7886

14A

Y78

8719

AY

7884

80B

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us

beot

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NW

130-

15E

CU

AD

OR

: Im

babu

ra, R

io C

ach

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AY

7886

15A

Y78

8720

AY

7884

81H

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W81

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OS

TA

RIC

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upp

lier

AY

7886

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Y78

8751

AY

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AY

7886

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Y78

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AY

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dir

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CO

ST

A R

ICA

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fly

farm

su

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AY

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96A

Y09

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7886

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Y78

8822

AY

7885

82S

myr

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RA

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Y78

8678

AY

7888

16A

Y78

8576

Nym

phal

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An

tan

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ZA

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Y78

8609

AY

7887

11A

Y78

8472

An

tan

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GA

ND

A: K

ibal

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ores

tA

Y78

8610

AY

7887

12A

Y78

8473

An

tan

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nia

NW

65-5

CA

ME

RO

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AY

2182

36A

Y21

8255

AF

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AY

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2762

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AY

2488

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F41

2784

Sym

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thia

hyp

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AN

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oun

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Y78

8680

AY

7888

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Y78

8578

Sym

bren

thia

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AN

: Kau

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un

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oun

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Y78

8679

AY

7888

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Y78

8577

Hyp

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bell

aP

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PE

RU

: Cu

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Y78

8638

AY

7887

57A

F24

6590

Hyp

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char

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W89

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EC

UA

DO

R: S

ucu

mbi

osA

Y78

8639

AY

7887

58A

Y78

8518

Hyp

anar

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kefe

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NW

89-6

EC

UA

DO

R: S

ucu

mbi

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Y78

8640

AY

7887

59A

Y78

8519

Hyp

anar

tia

leth

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W36

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RA

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: Min

as G

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sA

F18

7774

AY

7887

60A

Y78

8520

Hyp

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tia

lin

dig

iiP

E-0

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PE

RU

: Cu

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Qu

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da S

an L

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AY

2487

81A

Y24

8806

AF

4127

59V

anes

sa a

nn

abel

laN

W74

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SA

: Wyo

min

gA

Y78

8685

AY

7888

23A

Y78

8583

Van

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lan

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SW

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AY

0902

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Y09

0187

AF

4127

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anes

sa b

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NW

89-4

EC

UA

DO

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smer

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sA

Y78

8686

AY

7888

24A

Y78

8584

Van

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car

du

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W63

-3U

SA

: Mis

sou

riA

Y24

8782

AY

2488

07A

F41

2770

Van

essa

gon

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laN

W63

-4N

EW

ZE

AL

AN

DA

Y24

8784

AY

2488

09A

F41

2782

Van

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in

dic

aN

W63

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PA

NA

Y78

8687

AY

7888

25A

Y78

8585

Van

essa

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aN

W63

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NE

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EA

LA

ND

AY

7886

88A

Y78

8826

AY

7885

86V

anes

sa k

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awi

NW

77-3

AU

ST

RA

LIA

: Sou

th A

ust

rali

aA

Y78

8689

AY

7888

27A

Y78

8587

Van

essa

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inn

aN

W36

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RA

ZIL

: Min

as G

erai

sA

Y78

8690

AY

7888

28A

Y78

8588

Van

essa

vir

gin

ien

sis

NW

77-1

6U

SA

: Ten

nes

see

AY

2487

83A

Y24

8808

AY

2488

27A

glai

s io

NW

63-1

6S

WE

DE

N: S

tock

hol

mA

Y24

8785

AY

2488

10A

F41

2766

Agl

ais

mil

bert

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W77

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US

A: W

ash

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onA

Y24

8787

AY

2488

12A

Y24

8828

Agl

ais

urt

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NW

63-3

SW

ED

EN

: Sto

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AY

2487

86A

Y24

8811

AF

4127

77N

ymph

alis

an

tiop

aN

W70

-2S

WE

DE

N: S

tock

hol

mA

Y21

8246

AY

2182

66A

Y21

8284

Nym

phal

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alif

orn

ica

NW

74-1

4U

SA

: Ore

gon

AY

2487

89A

Y24

8814

AY

2488

30N

ymph

alis

l-a

lbu

mN

W78

-1C

AN

AD

A: B

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sh C

olu

mbi

aA

Y24

8791

AY

2488

16A

Y24

8832

Nym

phal

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hlo

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NW

62-2

SW

ED

EN

: Öla

nd

AY

2487

88A

Y24

8813

AY

2488

29N

ymph

alis

xan

thom

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NW

84-1

RU

SS

IA: Y

aku

tia

AY

2487

90A

Y24

8815

AY

2488

31P

olyg

onia

can

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EW

19-1

1JA

PA

NA

Y24

8792

AY

2488

17A

Y24

8833

Pol

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ia c

-alb

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NW

70-3

SW

ED

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: Sto

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AY

0902

22A

Y09

0188

AY

0901

54P

olyg

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c-a

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NW

65-8

JAP

AN

AY

2487

99A

Y24

8824

AF

4127

86P

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com

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65-6

US

A: T

enn

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eA

Y24

8794

AY

2488

19A

F41

2781

Pol

ygon

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NW

77-1

5G

RE

EC

EA

Y24

8800

AY

2488

25A

Y24

8838

Pol

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NW

74-1

2U

SA

: Ore

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AY

2487

98A

Y24

8823

AY

2488

37

Hig

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peci

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Loc

alit

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OI

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1-a

win

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s

AP

PE

ND

IX 1

Con

tin

ued



Pol

ygon

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di

NW

112-

3M

EX

ICO

: Son

ora,

Yec

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AY

7886

62A

Y78

8800

AY

7885

60P

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in

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US

A: T

enn

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eA

Y24

8793

AY

2488

18A

Y24

8834

Pol

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74-1

0U

SA

: Ore

gon

AY

7886

63A

Y78

8801

AY

7885

61P

olyg

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sat

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W74

-9U

SA

: Ore

gon

AY

2487

96A

Y24

8821

AY

2488

35P

olyg

onia

gra

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sN

W74

-6U

SA

: Ore

gon

AY

2487

97A

Y24

8822

AY

2488

36‘K

alli

min

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nar

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NW

68-5

CO

ST

A R

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: Bu

tter

fly

farm

su

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Y78

8606

AY

7887

08A

Y78

8469

An

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W66

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OS

TA

RIC

A: B

utt

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AY

7886

07A

Y78

8709

AY

7884

70A

nar

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W36

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RA

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: Min

as G

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Y78

8608

AY

7887

10A

Y78

8471

Sip

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64-7

CO

ST

A R

ICA

: Bu

tter

fly

farm

su

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Y78

8677

AY

7888

15A

Y78

8575

Sip

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W69

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TA

RIC

A: B

utt

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lier

AY

2182

48A

Y21

8268

AY

2182

86N

apeo

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ju

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NW

85-1

BR

AZ

IL: M

ato

Gro

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AY

7886

61A

Y78

8788

AY

7885

48M

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orph

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PE

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Ch

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Y78

8658

AY

7887

84A

Y78

8544

Hyp

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aN

W81

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US

TR

AL

IA: Q

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and

AY

7886

33A

Y78

8752

AY

7885

13H

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imn

as a

nth

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NW

68-8

KE

NY

A: B

utt

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y fa

rm s

upp

lier

AY

7886

34A

Y78

8753

AY

7885

14H

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as b

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aN

W62

-6A

US

TR

AL

IA: Q

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nsl

and

AY

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24A

Y09

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AY

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56H

ypol

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as m

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NW

68-3

KE

NY

A: B

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lier

AY

7886

35A

Y78

8754

AY

7885

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W80

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IND

ON

ES

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Isl

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AY

7886

36A

Y78

8755

AY

7885

16H

ypol

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NW

66-4

KE

NY

A: B

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AY

7886

37A

Y78

8756

AY

7885

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MA

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GA

SC

AR

: Man

drak

aA

Y78

8664

AY

7888

02A

Y78

8562

Pre

cis

anti

lope

NW

88-4

ZIM

BA

BW

E: M

aron

dera

AY

7886

65A

Y78

8803

AY

7885

63P

reci

s ar

ches

iaN

W88

-6Z

IMB

AB

WE

: Mar

onde

raA

Y78

8666

AY

7888

04A

Y78

8564

Pre

cis

cery

ne

NW

88-3

ZIM

BA

BW

E: M

aron

dera

AY

7886

67A

Y78

8805

AY

7885

65P

reci

s cu

ama

NW

83-1

3Z

IMB

AB

WE

: Har

are

AY

7886

68A

Y78

8806

AY

7885

66P

reci

s oc

tavi

aN

W68

-9K

EN

YA

: Bu

tter

fly

farm

su

ppli

erA

Y78

8669

AY

7888

07A

Y78

8567

Pre

cis

sin

uat

aN

W83

-1U

GA

ND

A: K

ibal

e F

ores

tA

Y78

8670

AY

7888

08A

Y78

8568

Pre

cis

tuge

laN

W11

4-15

ZA

MB

IA: N

of

Mw

inil

un

ga, L

esom

bo R

AY

7886

71A

Y78

8809

AY

7885

69R

hin

opal

pa p

olyn

ice

NW

81-5

MA

LA

YS

IAA

Y78

8674

AY

7888

12A

Y78

8572

Sal

amis

an

teva

NW

111-

9M

AD

AG

AS

CA

R: M

andr

aka

AY

7886

75A

Y78

8813

AY

7885

73S

alam

is c

acta

NW

82-3

UG

AN

DA

: Kib

ale

For

est

AY

7886

76A

Y78

8814

AY

7885

74Yo

ma

algi

na

NW

80-1

3P

NG

: Mor

obe

Pro

v., W

au V

alle

yA

Y78

8692

AY

7888

30A

Y78

8590

Pro

togo

nio

mor

pha

anac

ard

iiN

W73

-15

KE

NY

A: B

utt

erfl

y fa

rm s

upp

lier

AY

0902

23A

Y09

0189

AY

0901

55P

r. pa

rhas

sus

NW

82-7

UG

AN

DA

: Kib

ale

For

est

AY

7886

73A

Y78

8811

AY

7885

71P

r. cy

tora

NW

123-

23G

HA

NA

AY

7886

72A

Y78

8810

AY

7885

70Ju

non

ia a

rtax

iaN

W11

4-11

ZA

MB

IA: S

of

Mw

inil

un

gaA

Y78

8642

AY

7887

62A

Y78

8522

Jun

onia

coe

nia

NW

85-1

3U

SA

: Ten

nes

see

AY

7886

43A

Y24

8801

AY

2488

26Ju

non

ia i

phit

aN

W68

-17

Bu

tter

fly

farm

su

ppli

erA

Y09

0225

AY

0901

91A

Y09

0157

Jun

onia

eri

gon

eN

W81

-2P

NG

: Mor

obe

Pro

vin

ce, B

ulo

loA

Y78

8644

AY

7887

63A

Y78

8523

Jun

onia

nat

alic

aN

W68

-13

KE

NY

A: B

utt

erfl

y fa

rm s

upp

lier

AY

7886

45A

Y78

8764

AY

7885

24Ju

non

ia o

enon

eN

W68

-1K

EN

YA

: Bu

tter

fly

farm

su

ppli

erA

Y78

8646

AY

7887

65A

Y78

8525

Jun

onia

sop

hia

NW

83-1

0U

GA

ND

A: K

ibal

e F

ores

tA

Y78

8647

AY

7887

66A

Y78

8526

Jun

onia

ter

eaN

W68

-15

KE

NY

A: B

utt

erfl

y fa

rm s

upp

lier

AY

7886

48A

Y78

8767

AY

7885

27Ju

non

ia t

ouh

ilim

asa

NW

95-1

5Z

AM

BIA

: Kal

ene

Hil

lA

Y78

8649

AY

7887

68A

Y78

8528

Hig

her

tax

onS

peci

esV

ouch

er c

ode

Loc

alit

yC

OI

EF

1-a

win

gles

s



Kam

illa

cym

odoc

eN

W11

4-8

ZA

MB

IA: N

of

Mw

inil

un

ga, L

esom

bo R

AY

7886

52A

Y78

8771

AY

7885

31K

alli

ma

inac

hu

sN

W85

-15

Bu

tter

fly

farm

su

ppli

erA

Y78

8650

AY

7887

69A

Y78

8529

Kal

lim

a pa

rale

kta

NW

62-8

Bu

tter

fly

farm

su

ppli

erA

Y09

0229

AY

0901

97A

Y09

0163

Kal

lim

oid

es r

um

iaN

W96

-8G

HA

NA

: Ash

anti

Reg

ion

AY

7886

51A

Y78

8770

AY

7885

30C

atac

ropt

era

cloa

nth

eN

W88

-1Z

IMB

AB

WE

: Mar

onde

raA

Y78

8619

AY

7887

24A

Y78

8485

Mal

lika

jac

kson

iN

W12

2-6

TA

NZ

AN

IAA

Y78

8653

AY

7887

72A

Y78

8532

Dol

esch

alli

a bi

salt

ide

NW

64-5

Bu

tter

fly

farm

su

ppli

erA

Y78

8621

AY

7887

35A

Y78

8496

Van

essu

la m

ilca

NW

96-5

GH

AN

A: A

shan

ti R

egio

nA

Y78

8691

AY

7888

29A

Y78

8589

Mel

itae

ini

Eu

phyd

ryas

au

rin

iaN

W6-

4F

RA

NC

E: C

ervi

ères

AF

1877

46A

Y78

8743

AY

7885

04E

uph

ydry

as c

hal

ced

ona

NW

14-4

US

A: C

alif

orn

iaA

F18

7752

AY

7887

44A

Y78

8505

Eu

phyd

ryas

des

fon

tain

iiN

W70

-4S

PA

IN: E

l G

uix

AY

0902

26A

Y09

0193

AY

0901

59E

uph

ydry

as e

dit

ha

NW

5-8

US

A: C

alif

orn

iaA

F18

7765

AY

7887

45A

Y78

8506

Eu

phyd

ryas

gil

lett

iiN

W24

-6U

SA

: Mon

tan

aA

F18

7771

AY

7887

46A

Y78

8507

Eu

phyd

ryas

ph

aeto

nN

W13

-3U

SA

: Mar

ylan

dA

F18

7797

AY

7887

47A

Y78

8508

Ch

losy

ne

acas

tus

NW

35-1

5U

SA

: Col

orad

oA

F18

7735

AY

7887

25A

Y78

8486

Ch

losy

ne

cyn

eas

NW

38-1

7U

SA

: Ari

zon

aA

F18

7757

AY

7887

26A

Y78

8487

Ch

losy

ne

gau

dea

lis

NW

37-2

CO

ST

A R

ICA

: La

Sel

vaA

F18

7770

AY

7887

27A

Y78

8488

Ch

losy

ne

gorg

one

NW

34-4

US

A: C

olor

ado

AF

1877

72A

Y78

8728

AY

7884

89C

hlo

syn

e h

arri

sii

NW

35-1

0U

SA

: New

Yor

kA

F18

7773

AY

7887

29A

Y78

8490

Ch

losy

ne

jan

ais

NW

62-1

CO

ST

A R

ICA

: Bu

tter

fly

farm

su

ppli

erA

Y78

8620

AY

7887

30A

Y78

8491

Ch

losy

ne

laci

nia

NW

62-4

CO

ST

A R

ICA

: Bu

tter

fly

farm

su

ppli

erA

Y09

0227

AY

0901

95A

Y09

0161

Ch

losy

ne

nar

vaN

W37

-3C

OS

TA

RIC

A: L

a S

elva

AF

1877

86A

Y78

8731

AY

7884

92C

hlo

syn

e n

ycte

isN

W34

-5U

SA

: Col

orad

oA

F18

7788

AY

7887

32A

Y78

8493

Ch

losy

ne

pall

aN

W20

-4U

SA

: Cal

ifor

nia

AF

1877

91A

Y78

8733

AY

7884

94C

hlo

syn

e th

eon

aN

W27

-6U

SA

: Ari

zon

aA

F18

7808

AY

7887

34A

Y78

8495

Dym

asia

dym

asN

W27

-7U

SA

: Ari

zon

aA

F18

7764

AY

7887

85A

Y78

8545

Mic

roti

a el

vaN

W61

-1M

EX

ICO

: Ch

iapa

sA

Y78

8660

AY

7887

87A

Y78

8547

Tex

ola

elad

aN

W7-

1U

SA

: Tex

asA

Y78

8659

AY

7887

86A

Y78

8546

Pol

adry

as a

rach

ne

NW

27-4

US

A: C

alif

orn

iaA

F18

7740

AY

7887

99A

Y78

8559

Mel

itae

a ar

du

inn

aN

W23

-5G

RE

EC

E: P

isso

deri

AF

1877

42A

Y78

8774

AY

7885

34M

elit

aea

brit

omar

tis

NW

69-8

SW

ED

EN

AY

7886

55A

Y78

8775

AY

7885

35M

elit

aea

cin

xia

NW

73-1

4S

WE

DE

N: S

tock

hol

mA

Y78

8656

AY

7887

76A

Y78

8536

Mel

itae

a d

eion

eN

W95

-5F

RA

NC

E: A

ude

AY

7886

57A

Y78

8777

AY

7885

37M

elit

aea

did

ymoi

des

NW

26-1

RU

SS

IA: B

ury

atia

AF

1877

62A

Y09

0194

AY

0901

60M

elit

aea

lato

nig

ena

NW

25-3

RU

SS

IA: B

ury

atia

AF

1877

80A

Y78

8778

AY

7885

38

Hig

her

tax

onS

peci

esV

ouch

er c

ode

Loc

alit

yC

OI

EF

1-a

win

gles

s

AP

PE

ND

IX 1

Con

tin

ued



Mel

itae

a pe

rsea

NW

34-1

0L

EB

AN

ON

: Moh

afaz

at B

ehar

réA

F18

7796

AY

7887

79A

Y78

8539

Mel

itae

a pu

nic

aN

W34

-11

LE

BA

NO

N: M

ohaf

azat

Kes

ron

anA

F18

7803

AY

7887

81A

Y78

8541

Mel

itae

a sc

otos

iaN

W27

-11

CH

INA

: Heb

ei P

rovi

nce

AF

1878

04A

Y78

8780

AY

7885

40M

elit

aea

triv

iaN

W23

-6G

RE

EC

E: P

isso

deri

AF

1878

10A

Y78

8782

AY

7885

42M

elit

aea

vari

aN

W24

-13

FR

AN

CE

: Lau

s de

Cer

vièr

esA

F18

7812

AY

7887

83A

Y78

8543

Gn

ath

otri

che

excl

amat

ion

isN

W89

-9E

CU

AD

OR

: Su

cum

bios

AY

7886

29A

Y78

8748

AY

7885

09H

iggi

nsi

us

fasc

iatu

sP

E-1

0–20

PE

RU

: Cu

zco,

Qu

ebra

da C

hau

pim

ayo

AY

7886

30A

Y78

8749

AY

7885

10A

nth

anas

sa d

rusi

lla

NW

76-6

EC

UA

DO

R: E

smer

alda

sA

Y78

8611

AY

7887

14A

Y78

8475

An

than

assa

tex

ana

NW

12-6

US

A: T

exas

AF

1878

06A

Y78

8716

AY

7884

77A

nth

anas

sa a

rdys

NW

22-4

CO

ST

A R

ICA

: Mon

teve

rde

AF

1877

43A

Y78

8713

AY

7884

74A

nth

anas

sa o

tan

esN

W24

-4C

OS

TA

RIC

A: M

onte

verd

eA

F18

7790

AY

7887

15A

Y78

8476

An

than

assa

tu

lcis

NW

104-

12P

AN

AM

A: G

ambo

aA

Y78

8612

AY

7887

17A

Y78

8478

Cas

tili

a er

anit

esN

W76

-2E

CU

AD

OR

: Pic

hin

cha

AY

7886

17A

Y78

8722

AY

7884

83C

asti

lia

ofel

laN

W10

5-3

PA

NA

MA

: Ach

iote

Roa

dA

Y78

8618

AY

7887

23A

Y78

8484

Ere

sia

clio

NW

76-5

EC

UA

DO

R: E

smer

alda

sA

Y78

8622

AY

7887

36A

Y78

8497

Ere

sia

coel

aN

W10

4-3

PA

NA

MA

: Pat

h t

o G

lori

a A

lta

AY

7886

23A

Y78

8737

AY

7884

98E

resi

a eu

nic

eN

W92

-5B

RA

ZIL

: São

Pau

loA

Y78

8624

AY

7887

38A

Y78

8499

Ere

sia

leti

tia

NW

91-9

EC

UA

DO

R: S

ucu

mbi

osA

Y78

8625

AY

7887

39A

Y78

8500

Ere

sia

quin

till

aN

W76

-3E

CU

AD

OR

: Esm

eral

das

AY

7886

27A

Y78

8741

AY

7885

02E

resi

a pe

lon

iaN

W10

8-11

PE

RU

AY

7886

26A

Y78

8740

AY

7885

01E

resi

a se

stia

NW

76-8

EC

UA

DO

R: E

smer

alda

sA

Y78

8628

AY

7887

42A

Y78

8503

Jan

atel

la l

euco

des

ma

NW

85-1

6P

AN

AM

A: G

ambo

aA

Y78

8641

AY

7887

61A

Y78

8521

Maz

ia a

maz

onic

aN

W76

-6E

CU

AD

OR

AY

7886

54A

Y78

8773

AY

7885

33P

hyc

iod

es b

ates

iiN

W72

-4C

AN

AD

A: O

nta

rio

AF

1877

47A

Y78

8789

AY

7885

49P

hyc

iod

es c

ocyt

aN

W11

-4C

AN

AD

A: B

riti

sh C

olu

mbi

aA

F18

7755

AY

0901

92A

Y09

0158

Ph

ycio

des

myl

itta

NW

11-1

0C

AN

AD

A: B

riti

sh C

olu

mbi

aA

F18

7785

AY

7887

91A

Y78

8551

Ph

ycio

des

ors

eis

NW

67-3

US

A: C

alif

orn

iaA

Y15

6631

AY

7887

92A

Y78

8552

Ph

ycio

des

pal

lesc

ens

NW

64-2

ME

XIC

O: M

ich

oacá

nA

Y15

6640

AY

7887

93A

Y78

8553

Ph

ycio

des

pal

lid

aN

W34

-6U

SA

: Col

orad

oA

F18

7792

AY

7887

94A

Y78

8554

Ph

ycio

des

ph

aon

NW

35-1

1M

EX

ICO

: Maz

atla

nA

F18

7798

AY

7887

95A

Y78

8555

Ph

ycio

des

pic

taN

W34

-7U

SA

: Col

orad

oA

F18

7800

AY

7887

96A

Y78

8556

Ph

ycio

des

pu

lch

ella

NW

67-1

4U

SA

: Ore

gon

AY

1566

62A

Y78

8797

AY

7885

57P

hyc

iod

es t

har

osN

W34

-2U

SA

: Min

nes

ota

AF

1878

07A

Y78

8798

AY

7885

58P

hyc

iod

es g

raph

ica

NW

67-9

ME

XIC

O: M

exic

o S

tate

AY

1566

84A

Y78

8790

AY

7885

50T

elen

assa

tri

mac

ula

taN

W91

-6E

CU

AD

OR

: Su

cum

bios

AY

7886

83A

Y78

8821

AY

7885

81T

egos

a an

ieta

NW

91-1

1E

CU

AD

OR

: Su

cum

bios

AY

7886

81A

Y78

8819

AY

7885

79T

egos

a ti

ssoi

des

NW

76-4

EC

UA

DO

R: E

smer

alda

sA

Y78

8682

AY

7888

20A

Y78

8580

Hig

her

tax

onS

peci

esV

ouch

er c

ode

Loc

alit

yC

OI

EF

1-a

win

gles

s



APPENDIX 2

Proposed higher classification of the subfamilyNymphalinae based on results presented in this paper.Genera marked with asterisks were not sampled forthis study. For a full synonymic list of the speciesplease see http://www.zoologi.su.se/research/wahlberg

Subfamily NYMPHALINAE Rafinesque, 1815incertae sedis

Pycina* Doubleday, 1849Kallimoides Shirôzu & Nakanishi, 1984Vanessula Dewitz, 1887Rhinopalpa Felder & Felder, 1860

Tribe Coeini Scudder, 1893Historis Hübner, 1819

= Coea Hübner, 1819= Aganisthos Boisduval & Le Conte, 1835= Megistanis Doubleday, 1845= Megistanis Boisduval, 1870

Baeotus Hemming, 1939Tribe Nymphalini Rafinesque, 1815

Colobura Billberg, 1820= Gynoecia Doubleday, 1845

Tigridia Hübner, 1819= Callizona Doubleday, 1848

Smyrna Hübner, 1823Antanartia Rothschild & Jordan, 1903Araschnia Hübner, 1819Symbrenthia Hübner, 1819

= Laogona Boisduval, 1836= Brensymthia Huang 2001

Mynes Boisduval, 1832Hypanartia Hübner, 1821

= Eurema Doubleday, 1845Vanessa Fabricius, 1807

= Nymphalis Latreille, 1804= Cynthia Fabricius, 1807= Pyrameis Hübner, 1819= Bassaris Hübner, 1821= Ammiralis Rennie, 1832= Neopyrameis Scudder, 1889= Fieldia Niculescu, 1979

Aglais Dalman, 1816= Ichnusa Reuss, 1939= Inachis Hübner, 1819= Hamadryas Hübner, 1806

Nymphalis Kluk, 1780= Scudderia Grote, 1873= Euvanessa Scudder, 1889= Roddia Korshunov, 1996

Polygonia Hübner, 1819= Kaniska Moore, 1899= Eugonia Hübner, 1819= Comma Rennie, 1832= Grapta Kirby, 1837

Tribe Victorinini Scudder, 1893Anartia Hübner, 1819

= Celaena Doubleday, 1849= Celoena Boisduval, 1870= Anartiella Fruhstorfer, 1907

Siproeta Hübner, 1823= Victorina Blanchard, 1840= Amphirene Doubleday, 1845= Amphirene Boisduval, 1870= Aphnaea Capronnier, 1881

Napeocles Bates, 1864Metamorpha Hübner, 1819

Tribe Junoniini Reuter, 1896Junonia Hübner, 1819

= Alcyoneis Hübner, 1819= Aresta Billberg, 1820= Kamilla Collins & Larsen, 1991

Salamis Boisduval, 1833Yoma Doherty, 1886

= Yoma de Nicéville, 1886Protogoniomorpha Wallengren,

1857Precis Hübner, 1819

= Coryphaeola Butler, 1878= Kallimula Holland, 1920

Hypolimnas Hübner, 1819= Esoptria Hübner, 1819= Diadema Boisduval, 1832= Euralia Westwood, 1850= Eucalia Felder, 1861

Tribe Kallimini Doherty, 1886Kallima Doubleday, 1849Doleschallia Felder & Felder, 1860

= Apatura Hübner, 1819Catacroptera Karsch, 1894Mallika Collins & Larsen, 1991

Tribe Melitaeini Newman, 1870incertae sedis

Antillea* Higgins, 1959Atlantea* Higgins, 1959Gnathotriche Felder & Felder, 1862

= Gnathotrusia Higgins, 1981Higginsius Hemming, 1964

= Fulvia Higgins, 1959Chlosyne Butler, 1870

= Morpheis Geyer, 1833= Synchloe Doubleday, 1845= Anemeca Kirby, 1871= Coatlantona Kirby, 1871= Limnaecia Scudder, 1872= Charidryas Scudder, 1872= Thessalia Scudder, 1875

Microtia Bates, 1864Texola Higgins, 1959Dymasia Higgins, 1960Poladryas Bauer, 1961




Subtribe Euphydryina Higgins, 1978Euphydryas Scudder, 1872

= Lemonias Hübner, 1806= Occidryas Higgins, 1978= Eurodryas Higgins, 1978= Hypodryas Higgins, 1978

Subtribe Melitaeina Newman, 1870Melitaea Fabricius, 1807

= Lucina Rafinesque, 1815= Schoenis Hübner, 1819= Cinclidia Hübner, 1819

= Mellicta Billberg, 1820= Didymaeformia Verity, 1950

= Athaliaeformia Verity, 1950

Subtribe Phyciodina Higgins, 1981Phyciodes Hübner, 1819Phystis* Higgins, 1981Anthanassa Scudder, 1875

= Tritanassa Forbes, 1945Dagon* Higgins, 1981Telenassa Higgins, 1981Ortilia* Higgins, 1981Tisona* Higgins, 1981Tegosa Higgins, 1981Eresia Boisduval, 1836Castilia Higgins, 1981Janatella Higgins, 1981Mazia Higgins, 1981

phylogenetic relationships and historical biogeography of tribes … · (lepidoptera: nymphalidae)...

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