arthropod structure & developmentigarzon.weebly.com/.../simonsen_etal_2012_review.pdf · 310 t.j....

at SciVerse ScienceDirect

Arthropod Structure & Development 41 (2012) 307e322

Contents lists available

Arthropod Structure & Development

journal homepage: www.elsevier .com/locate/asd

Butterfly morphology in a molecular age e Does it still matter in butterflysystematics?

Thomas J. Simonsen a,*, Rienk de Jong b, Maria Heikkilä c, Lauri Kaila c

aDepartment of Life Sciences, Natural History Museum, Cromwell Road, London SW7 5BD, United KingdombNetherlands Centre for Biodiversity Naturalis, P.O. Box 9517, 2300 RA Leiden, The Netherlandsc Finnish Museum of Natural History, Zoology Unit, P.O. Box 17, FI-00014 University of Helsinki, Finland

a r t i c l e i n f o

Article history:Received 22 December 2011Received in revised form30 March 2012Accepted 23 April 2012

Keywords:ButterfliesPhylogenyMorphologyMolecularCharacter evolution

* Corresponding author. Tel.: þ44 (0)20 7942 6548E-mail addresses: [email protected] (T.J.

ncbnaturalis.nl (R. de Jong), [email protected] (L. Kaila).

1467-8039/$ e see front matter Crown Copyright � 2doi:10.1016/j.asd.2012.04.006

a b s t r a c t

We review morphological characters considered important for understanding butterfly phylogeny andevolution in the light of recent large-scale molecular phylogenies of the group. A number of the mostimportant morphological works from the past half century are reviewed and morphological characterevolution is reassessed based on the most recent phylogenetic results. In particular, higher level butterflymorphology is evaluated based on a very recent study combining an elaborate morphological datasetwith a similar molecular one. Special attention is also given to the families Papilionidae, Nymphalidaeand Hesperiidae which have all seen morphological and molecular efforts come together in large,combined works in recent years. In all of the examined cases the synergistic effect of combining elab-orate morphological datasets with ditto molecular clearly outweigh the merits of either data type ana-lysed on its own (even for ‘genome size’ molecular datasets). It is evident that morphology, far frombeing obsolete or arcane, still has an immensely important role to play in butterfly (and insect) phylo-genetics. Not least because understanding morphology is essential for understanding and evaluating theevolutionary scenarios phylogenetic trees are supposed to illustrate.

Crown Copyright � 2012 Published by Elsevier Ltd. All rights reserved.

1. Introduction

Evolution has left traces of its history in every character, be itmorphological, molecular, physiological, ethological or whatever.Thus, studying any set of characters unmasks a bit (tiny as it maybe) about evolutionary history. Ideally, all different sets of charac-ters should tell the same story for the group under study, but we donot live in an ideal world and different sets of characters quite yieldconflicting results. A single set of characters may also seem to telldifferent stories about evolution. There are several ways, non-statistical like Maximum Parsimony or statistical like Bayesiananalyses, to make a choice between conflicting phylogenetic trees,but there is no method to decide between conflicting results basedon different sets of characters. Each time a new set of characters isdiscovered and studied, hopes raise high that this will give the finalanswer. This was the case with ontogeny, the large scale applicationof the study of genital appendages, ultrastructures unveiled by theelectron microscope, and the rapid development of techniques for

; fax: þ44 (0)20 7942 5229.Simonsen), rienk.dejong@

.fi (M. Heikkilä), lauri.kaila@

012 Published by Elsevier Ltd. All

DNA sequencing. So far, such hopes have been overoptimistic, andafter some time each new set of characters explored been proven tohave its own flaws and drawbacks. Combining different datasets(such as morphological and molecular) will often yield more stableend results and allow for close scrutiny of conflicts between sub-datasets (e.g. Simonsen et al., 2006; Warren et al., 2009; Heikkiläet al., 2012) and thus to some extent assist decisions betweendata conflicts. Perhaps the best way to proceed in case of conflictsbetween different sets of characters is to analyse the conflictingevidence in terms of evolutionary impact.

Butterflies are perhaps the best studied higher group of inver-tebrate organisms and their systematics has been the subject of animmense bodyof studies forwell over a century. Despite comprisingonly a fewwell-defined families (e.g. Ackery et al., 1999) and a rela-tive lownumber of species for amega-diverse group (18,771 speciesin 1815 genera, vanNieukerken et al., 2011, Fig.1), the group’s higherphylogeny (family and subfamily level) has proved to be remarkablyhard to resolve. In the present paper a far fromexhaustive account isgiven of the most influential and important contributions since themiddle of the last century, not only to show the progress in the studyof butterfly systematics, but also to find where morphological andmolecular evidence agree, andwhere they seem to be in conflict andwhat could be done about it.

rights reserved.

Fig. 1. Examples of diversity of the butterfly families. AeC: Papilionidae. DeF: Hesperiidae. G: Hedylidae. HeJ: Pieridae. KeM: Nymphalidae. NeP: Lycaenidae. QeS: Riodinidae.

T.J. Simonsen et al. / Arthropod Structure & Development 41 (2012) 307e322 309

2. Higher butterfly phylogeny

2.1. Ehrlich et al. (1958e1967)

Paul R. Ehrlich in a series of publications (somewith co-authors)gave the first truly thorough account of the comparativemorphology and anatomy of Rhopalocera (¼Papilionoidea sensuvan Nieukerken et al., 2011) and its implications for higher-levelclassification of butterflies in the strict sense (¼Papilionoideasensu Ehrlich). While a number of these publications are morpho-logical accounts without an explicit systematic component (i.e.Ehrlich, 1960, 1961; Ehrlich and Davidson, 1961; Ehrlich andEhrlich, 1962, 1963 e although character matrices are given in thetwo latter), Ehrlich (1958) and Ehrlich and Ehrlich (1967) stand outas some of the most important and influential publications onhigher-level butterfly systematics in the 20th century. Ehrlich(1958) dissected adults from approximately 300 species of butter-flies in 240 genera representing all then-recognised subfamilies. Hetreated each body region in detail and identified 64 systematiccharacters and, by comparing them to representatives of 24 familiesof moths and skippers, made a subjective assessment of theprimitive and specialized state of each. Although some characterswere vaguely defined (i.e. “size moderate” vs. “size extreme”, or“dull colored” vs. “brightly colored”) many characters (in particularthoracic and head characters) were examined and illustrated forthe first time. The resultant phylogeny was necessarily not cladisticin the strict sense (Hennig’s methodology was after all not availablein English until 1966), but since it was based on an assessment ofprimitive and specialized character states, it is certainly fair to call it“proto-cladistic” (see also Vane-Wright, 2003). In the phylogeny,Lycaenidae (including Riodinidae) was based in a basal polytomywith (Nymphalidaeþ Libytheidae) and (Papilionidaeþ Pieridae).Ehrlich and Ehrlich (1967) published classification of butterflies(including skippers) based on numerical phenetic analyses of thecharacter sets established in the previous 10 years of publications(listed above). Although they found some support for Ehrlich’s(1958) relationships, they found that the result was dependant onthe analytical method and concluded that archiving a “general”classification was an impossibility. Despite being analyticallyoutdated today, Ehrlich’s contributions are still of singularlyimportance today as reference work for Lepidoptera morphology.

2.2. Kristensen (1976)

Kristensen (1976) stands as the first truly cladistic treatment ofhigher-level butterfly systematics and as such as one of the mostimportant contributions to the understanding of butterflyphylogeny (Fig. 2A). The study was to a high degree based oncladistic re-analyses of Ehrlich’s results (from the publications listedabove), but characters fromother sources (i.e. Ehnbom,1948;Hessel,1966, 1969; Brock, 1971) were also considered. He identified fourprobable synapomorphies for Hesperioidea plus Papilionoidea:

1. Mesal fusion of dorsal laminae of secondary metafurcal arms.2. Butterfly type brain with very large optical lobes and small

deutocerebrum.3. Mesothoracic aorta with horizontal chamber.4. Oblique lateral dorsal muscle in mesothorax twisted.

He expressed some reservation with respect to the last twocharacters since 3 is absent in Papilionidae and 4 is absent inLycaenidae. In both cases he considered the absence of the char-acter to be secondary.

He further identified three potential autapomorphies forHesperioidea:

1. Forewing with no peripheral veins stalked.2. Forewing with CuP absent.3. Antennae with subapical thickenings.

And two for Papilionoidea:

1. Retinaculo-frenate wing coupling lost in both sexes.2. Antennae with apical clubs.

Some of these characters are problematic since CuP is lost in allother Papilionoidea than the family Papilionidae where it isretained as a short spur; and the retinaculo-frenate wing couplingis lost in all hesperioids except for male Euschemon (see below).Despite this relatively weak support, both superfamilies wereretained as working hypotheses.

Within Papilionoidea, Kristensen suggested that Papilionidaewere the sister group of a clade comprising the remaining threefamilieswith Pieridae being the sister of Nymphalidaeþ Lycaenidae(including Riodinidae). Four probable synapomorphies for Pieridae,Nymphalidae, and Lycaenidae were identified:

1. Loss of foreleg epiphysis.2. Maxillary palpus unisegmented or absent.3. Loss of prospinasterno-procoxal muscle.4. Presence of a secondary sclerite behind metascutellum.

Character 2 is found in Papilionidae as well except the basalgenus Baronia, but apart from that the character support for theclade appeared high.

The sister group relationship between Nymphalidae and Lycaen-idae was found to be supported by three constant characters:

1. Forelegs reduced in males and antennal cleaning performed bymiddle legs in both sexes.

2. Mesothoracic precoxal sulcus (¼suture) present.3. Prothoracic “presternum” present.

2.3. Scoble (1986)

Scoble (1986) suggested the first major change to the concept ofbutterflies for the better part of half a century when he suggestedthat the South American ‘moth’ family Hedylidae (with only thegenus Macrosoma e Fig. 1G) in reality belongs to Rhopalocera.Hedylidaewas described by Guenée for three small genera (all to besynonymized with Macrosoma by Scoble and earlier workers) of‘geometrid-like moths’ from South America, but was at the timeconsidered a tribe in the geometrid subfamily Oenochrominae (seeScoble, 1986 for details). As Scoble (1986) demonstrated, not onlydo Hedylidae not share any apomorphies with Oenochrominae orany other group of Geometridae, the family shares several potentialsynapomorphies with Rhopalocera. Scoble discussed 13 charactersthat supported not only the inclusion of Hedylidae in Rhopalocera,but also potentially a sister group relationship between Hedylidaeand Papilionoidea.

Potential synapomorphies for Rhopalocera incl. Hedylidae:

1. Apophyses of metathoracic furca sagittate.2. Pupa girdled.3. Second median plate of forewing lies partly under the base of

vein 1A.4. Larva with anal comb.5. Postspiracular bar on the first abdominal segment (Fig. 3AeB).6. Anterior apophyses in female genitalia reduced.

Fig. 2. Major butterfly phylogeny hypotheses since the advent of cladistics methodology. A: Kristensen (1976) (“the classical hypothesis”). B: The first major exemplar based studies;de Jong et al. (1996) (morphology), and Wahlberg et al. (2005) (molecular) included Hedylidae, but otherwise agreed with Kristensen. C: Butterfly relationships as found in thelarge-scale molecular Lepidoptera study by Regier et al. (2009). D: The identical (at the family level) results of the large-scale molecular Lepidoptera study by Mutanen et al. (2010)and the combined butterfly study by Heikkilä et al. (2012).

T.J. Simonsen et al. / Arthropod Structure & Development 41 (2012) 307e322310

Potential synapomorphies for Hedylidae and Papilionoidea:

1. Abdomen curved, particularly so in males.2. Abdomen T1 strongly pouched.3. Precoxal sulcus joins marginopleural sulcus.4. Pupal cocoon lost.5. Loss of temporal cleavage line in pupa.

6. Crochets of ventral prolegs of larva not forming a complete circle.7. Loss of pretarsus in the foreleg of male.

Scoble (1986) acknowledged that several of these characterswere either weak or known only from few specimens (e.g. thecocoon is present in several subordinate Hesperiidae and Papil-ionidae). However, the placement of Hedylidae in Rhopalocera has

Fig. 3. The post spiracular bar of abdomen 1 (arrows) considered an autapomorphy ofbutterflies including Hedylidae. A: Hedylidae (Macrosoma lucivittata). B: Nymphalidae(Tanaecia orphne). Although the anterior (tergum 1) component is more stronglymelanised, the composite (tergum 1þ2) nature of the bar is clearly recognisable.


since gained wide acceptance despite some initial critique(Weintraub and Miller, 1987).

2.4. Scott and Wright (1990)

Scott and Wright (1990) gave an overview of butterflyphylogeny and morphology (based on Scott’s 1985 study) inKudrna’s Butterflies of Europe (Kudrna, 1990). The overall rela-tionships found were similar to those of Ehrlich, i.e. Hesperioideaand Papilionoidea were found to be sister taxa, within Papil-ionoidea Papilionidae was found to be the sister of Pieridae. Scottand Wright (1990) rejected Scoble’s redefined concept of butter-flies and considered Rhopalocera to be comprised by Hesperioideaand Papilionoidea only, but accepted that Rhopalocera and Hedyl-idae comprised a monophyletic group.

The exclusion of Hedylidae from Rhopalocera was based partlyon the fact that Hedylidae lack two classical butterfly recognitioncharacters (clubbed antennae, and the absence of frenulum/reti-nacular wing coupling); and partly on a series of characters thatwere either homoplastic, erroneously polarised (based on theassumption that butterflies were Macrolepidoptera), or poorlyknown for Hedylidae.

Some of the latter (e.g. egg shape, a chambered aorta) have sincebeen found in Hedylidae as well as in Hesperioidea and Papil-ionoidea. Despite being somewhat subjective and not includinga rigorous analysis of the data, Scott andWright (1990) must still beconsidered an important contribution since it offers a good over-view of potentially important characters for butterfly systematics.

2.5. de Jong et al. (1996)

de Jong et al. (1996) presented the first exemplar basedcomputer cladistic study of butterfly phylogeny (summed up withsome additional observations by the same authors in Ackery et al.1999). Based on a dataset comprising 59 species of butterflies and15 moths, and 103 morphological characters they explored thephylogenetic relationships of Rhopalocera (sensu Scoble, 1986) atthe family and subfamily levels. They listed 10 potential synapo-morphies for Hedylidae, Hesperioidea and Papilionoidea, none ofwhich was universal or unique across the three groups, and onlythree were included in their final character matrix. Compared toScoble (1986) the most important results here were: confirmationthat Hedylidae have a horizontal chamber in the mesothoracicaorta (fromMinet,1988; Scoble and Aiello,1990); confirmation thatthe dorso-lateral postspiracular bar between A1 and A2 is struc-turally different from the condition found in some Axioidea(¼Cimelioidea) and Geometroidea; mesoscutum with secondaryline of weakness near median notal wing process (after Minet,1991); and apophyses of metathoracic furca sagittate. Despitehigh levels of homoplasy in the dataset, they concluded that Rho-palocera were most likely monophyletic, and recovered the “clas-sical” family-level relationships as the most likely workinghypothesis (Fig. 2B). A result here that will be shown to be ofpotentially great importance is the confirmation of the horizontalaorta chamberwhich is absent in Papilionidae.Within Rhopalocera,they concluded that Papilionoidea and Hesperioidea were sistertaxa based on six potential synapomorphies, of which only twowere considered entirely reliable (Ackery et al., 1999): reducedmesothoracic anepisternum (Fig. 4), and 2nd median plate offorewing base beneath vein 1Aþ 2A (Fig. 5BeC). They listed fiveuniversal thoracic characters in support for a monophyletic Papil-ionoidea, but were unable to find clear character support for eithera sister group relationship between Papilionidae and Pieridae orPieridae and a clade comprising Nymphalidae and Lycaenidae(including Riodinidae), but accepted the latter arrangement asa working hypothesis.

2.6. Wahlberg et al. (2005)

Wahlberg et al. (2005) presented the first combined higher-levelphylogenetic analyses of butterflies based on both morphologicalcharacters (from de Jong et al., 1996) andmultiple genes (in all 3159bp from COI, Elongation Factor 1 Alpha, andWingless). When all datawere analysed together the results supported the superfamily andfamily level results of de Jong et al. (1996) (Fig. 2B).

2.7. Higher level, large scale molecular studies

In 2009e2010 two studies published in short succession focusedon family-level Lepidoptera phylogenetics based on severalmolecular (primarily nuclear) markers, and both presenting novelresults with respect to the family level relationships of butterflies.Regier et al. (2009), in analyses of approximately 6800 bp from 5protein coding nuclear genes found that Rhopalocera were poly-phyletic (albeit with weak support) with Papilionidae being thesister group of the small macro moth family Callidulidae, anda clade comprising Hesperiidae and Hedylidae being the sistergroup of the micro moth family Thyrididae. The sister group of thelatter clade was a clade comprising the non-papilionid Papil-ionoidea (in a traditional arrangement), and that entire clade inturnwas the sister of Callidulidaeþ Papilionidae (Fig. 2C). Mutanenet al. (2010) in analyses of 4451 bp (3rd base removed from thedataset for most genes) from 7 protein coding nuclear and onemitochondrial gene also found that Thyrididae and Callidulidae

Fig. 4. Transformation of the anepisternum (shaded in grey, marked by arrows). A: Fully developed anepisternum in a Cossidae moth (Cossus). B: The somewhat reduced ane-pisternum in lower Hesperiidae and Hedylidae (Hesperiidae: Euschemon). C: The more reduced anepisternum found in higher Hesperiidae (Hesperia). D: The highly reducedanepisternum found in all other butterflies (Pieridae: Eurema). B and C redrawn after de Jong et al. (1996).

Fig. 5. Wing base sclerites showing the position of the 2nd medial plate (dark grey) and the shape of the 3rd axilary sclerite (light grey). A: The unmodified 3rd sclerite and fullyvisible 2nd plate in a Geometridae moth (Biston betularia). B: The unmodified 3rd sclerite and 2nd plate partly under the base of vein 1A in Pieridae (Pseudopontia paradoxa). C: Thetriangular unmodified 3rd sclerite and 2nd plate partly under the base of vein 1A in Hesperiidae (Allora doleschallii). D: The triangular unmodified 3rd sclerite and 2nd plate fullyobscured under the base of vein 1A in Hedylidae (Macrosoma semiermis).


Fig. 6. Schematic illustration of the mesophragmatic ridges (arrows). A: The flat ridgesfound in Hesperiidae and Hedylidae. B: The high ridges found in other butterflies.


were closely related to Rhopalocera, but the last was found to bemonophyletic with good support (Fig. 2D). However, like Regieret al. (2009), Mutanen et al. (2010) did find that Hesperiidae andHedylidae were sister taxa, and that Papilionoidea were non-monophyletic with the non-papilionid Papilionoidea being thesister group of HesperiidaeþHedylidae. Since both studies werebased solely on molecular data, neither discussed the morpholog-ical implications of these new results.

2.8. Heikkilä et al. (2012)

In a very recent study, Heikkilä et al. (2012) focused on thephylogeny of butterflies and analysed a dataset comprised by 6165bp from the genes used by Mutanen et al. (2010) together with 191mainly skeletal morphological characters from all life stages. In all,24 outgroup taxa and 54 ingroup (Rhopalocera) species wereincluded in their dataset. The results supported those of Mutanenet al. (2010) with Rhopalocera being monophyletic, but Papil-ionoidea being paraphyletic with HesperiidaeþHedylidae as thesister group of the non-papilionid Papilionoidea (Fig. 2D). Allhigher-level relationships received good support in at least some ofthe analyses. Since Heikkilä et al. (2012) included a largemorphological dataset in their study, their results provided anexcellent opportunity to examine morphological implications ofthe rearrangement of butterflies. This will be done in the nextsection.

2.9. Morphological consequences

The three recent studies listed above all agreed on two crucialand controversial issues: 1) Hesperiidae and Hedylidae are sistertaxa; and 2) Papilionoidea are non-monophyletic. In the followingwe will base our discussion on the Mutanen et al. (2010) andHeikkilä et al. (2012) result where Rhopalocera is monophyletic,but Papilionidae are the sister group of (Hesperi-idaeþHedylidae)þ non-papilionid Papilionoidea. The morpho-logical dataset Heikkilä et al. (2012) was traced on the phylogenyusing Mesquite (Maddison and Maddison, 2011).

2.9.1. Monophyly of RhopaloceraHeikkilä et al. (2012) demonstrated that several morphological

characters from both immature and adult life stages support themonophyly of Rhopalocera with Callidulidae and Thyrididaeexcluded (their outgroup dataset comprised several species fromeach of these two families). Three adult characters in particularwere found to be in support of a monophyletic Rhopalocera:

1. Metathoracic furca with ventro-distal extension. Althoughdifferently phrased, this character has been considereda butterfly autapomorphy since the study of Brock (1971).

2. Median plate 2 of forewing partly covered by base of vein 1Aþ 2A(Fig. 5BeD). This too is a classical butterfly autapomorphy,althoughde Jonget al. (1996) andAckeryet al. (1999) considered itto be a synapomorphy for Hesperiidaeþ Papilionoidea.

3. Abdominal tergum 1 domed. Again a classical autapomorphy,that varies somewhat between taxa.

Several immature characters similarly supported the mono-phyly of Rhopalocera, of which the two following appear to be ofparticular importance:

1. Pupal mesothorax laterally swollen. This appears unique to Rho-palocera including Hedylidae, and is universal in Rhopalocera.

2. Silken girdle supporting pupa: developed a few times in Lepi-doptera in general (Elachistinae in Gelechioidea, Cyclophorini

in Geometridae). A ‘groundplan’ character for Rhopalocera,retained even in some Hesperiidae that pupate internally ina cocoon. Reversed and thus absent in Nymphalidae and someLycaenidae (e.g. Scott and Wright, 1990).

The post spiracular bar (Fig. 3) was found to be difficult tohomologize across Rhopalocera by Heikkilä et al. (2012), and thusomitted from their analyses. The structure can be obvious in sometaxa as illustrated in Fig. 3, but very hard to identify with confi-dence in others. It is clear that this structure needs closer exami-nation in the future.

2.9.2. Sister group relationship between Hesperiidae and HedylidaeThe sister group relationship between Hesperiidae and Hedyl-

idae was recovered in all three studies with moderate support inRegier et al. (2009), and strong to very strong support in Mutanenet al. (2010) and Heikkilä et al. (2012). In previous studies no syn-apomorphies for the two families have been suggested, butHeikkilä et al. (2012) did find two unique synapomorphies thatunite Hesperiidae and Hedylidae:

1. FW axillary 3 irregularly triangular (Fig. 5CeD).2. Mesophragma with flat ridges (Fig. 6A).

Both character states were previously thought to be an aut-apomorphy of Hesperiidae (e.g. de Jong et al., 1996), but Heikkiläet al. (2012) demonstrated their presence in Hedylidae as well.The ridges of mesophragma in Hesperiidae and Hedylidae clearlydiffer from the high, often pointed ridges found in other butterflies(Fig. 6B). Despite these new characters, the positions of Hesperiidaeand Hedylidae are not unproblematic. At least three charactersrelated to the antenna could be seen as synapomorphies for a sistergroup relationship between Hesperiidae and Papilionoidea:

1. Antenna flagellum clubbed.2. Scape globular.


3. Base of scape sunken into head.

In Hedylidae the flagellum is variably thread-like or evenpectinate, but never clubbed, and the scape is barrel shaped and notsunken into the head. However, there are differences betweenHesperioidea and Papilionoidea as well (as already noted byKristensen, 1976), since the club is subapical in the former andapical in the latter. Furthermore, the clubbed antenna is found inseveral other day active Lepidoptera, and seems to be stronglylinked to that particular life style and probably not of particularsystematic value (Heikkilä, Kaila, and Simonsen, in preparation).

Only one character was found by Heikkilä et al. (2012) to bea potential synapomorphy of Hedylidae and Papilionoidea:

1. Pronotum with lateral dorsal plate processes.

2.9.3. Non-monophyly of PapilionoideaThe non-monophyly of Papilionoidea is beyond doubt the most

controversial result in all three studies. And even in the dataset ofHeikkilä et al. (2012) three morphological characters are univer-sally found in Papilionoidea, but not in Hedylidae or Hesperioidea,and would thus support a monophyletic Papilionoidea:

1. Mesothoracic anepisternum reduced to tiny sclerite or absent(Fig. 4D).

2. Membrane attaching tegulae to mesonotum at ventral edge.3. Precoxal sulcus not reaching anterior margin of basisternum.

The first character is somewhat problematic since the anepi-sternum is reduced to some extent in Hesperiidae (Fig. 4BeC) aswell; in fact there is even a continous reduction within that familyas evident by the larger anepisternum found in Euschemon (Fig. 4B)compared to Hesperiinae (Fig. 4C). However, the anepisternum inHesperiidae is never reduced to the extreme degree found in Pap-ilionoidea (Fig. 4D). Ackery et al. (1999) listed three additionalcharacters supporting a monophyletic Papilionoidea: secondarysclerite present behind metascutellum; parepisternal suturerunning in a straight or smoothly curved line; and mesophragmawith dorsal processes. The first character could not be verified withconfidence by Heikkilä et al. (2012) and was thus omitted from thatstudy, whereas the last two were interpreted or scored different toAckery et al. (1999). Heikkilä et al. (2012) found the parepisternalsuture in Hedylidae to differ from that in Hesperiidae and Lycae-nidae, but to be similar to the condition found in some Nympha-lidae and Papilionidae. The processes of the mesophragma werediscussed by Heikkilä et al. (2012), but polarised differently becauseof the signal in the molecular dataset. The mesophragmal processes(Fig. 6B) were indeed found in all classic Papilionoidea families, butis better interpreted as a plesiomorphic condition which is modi-fied to the flat ridges found in Hesperiidae and Hedylidae. Themesophragmal ridges can thus been seen as a good example of howstrong molecular results can lead to re-evaluation of the evolutionof morphological characters.

The morphology is clearly ambiguous with respect to the posi-tion of Papilionidae, and when morphology was analysed alone,Heikkilä et al. (2012) were unable to recover a well-supportedphylogeny for the butterfly families. Based on Heikkilä et al.’s(2012) dataset only three non-universal wing characters tenta-tively support a sister group relationship between Hesperi-idaeþHedylidae and the non-papilionid Papilionoidea:

1. Forewing spur (CuP) absent.2. Forewing: Outer margin of axillary 3: with blunt tooth on

which base of vein 1Aþ 2A hinges.

3. Hind wing with median arm reduced.

The first character is scored as universal in all non-papilionidRhopalocera by Heikkilä et al. (2012). This is, however, contra-dicted by Scoble (1986) where a vestigial CuP is illustrated infig. 32a. However, this vein is shown as present in its entire length,but very weakly developed, and thus very different from the short,but stout spur which represents CuP in Papilionidae (e.g. Miller,1987, Fig. 7D). Re-examination of Hedylidae wing preparationsin the collections of NHM (BM(NH) Geom. 11861, 11884, 11963,12044) and the Finnish Museum of Natural History furtherrevealed that while a structure corresponding to Scoble’s fig. 32ais absent in some species (Fig. 7A) but present in others(Fig. 7BeC), it may be a fold and not the vestiges of a vein. Indeed,it appears to be similar in structure to a longitudinal fold foundinside the discal cell in the same specimens. It furthermore differsfrom the spur found in Papilionidae since it originate between thebase of the Cu-stem and 1A (Fig. 7C) and not on the Cubital-stemas in Papilionidae (Fig. 7D). The second character is reversed inLycaenidae and Riodinidae, and the third is reduced in someHesperiidae. However, one character not included by Heikkiläet al. (2012) potentially yields strong support for the sistergroup relationship between HesperiidaeþHedylidae and thenon-papilionid Papilionoidea:

- Mesothoracic aorta with horizontal chamber. This characterhas long been known to be absent from Papilionidae, butpresent in Pieridae, Nymphalidae, Lycanidae and Hesperiidae(e.g. Kristensen, 1976), and has been confirmed for Hedylidae(Minet, 1988; Scoble and Aiello, 1990). Heikkilä et al. (2012) didnot include soft tissue characters for practical reasons, and thecharacter has not been confirmed across a broad sample ofspecies and genera for the same reason. Nevertheless, it seemsreasonable that it at least belongs to the ground plan ofa putative clade comprising Pieridae, Nymphalidae, Lycanidae,Hesperiidae and Hedylidae; but excluding Papilionidae.Combined with the strong molecular results and the three lessstrong characters listed above, this provides considerablesupport for a redefinition of Rhopalocera where Papilionidae isthe sister of the remaining families.

3. Papilionidae

This family, the true swallowtails, has probably been thesubject of more phylogenetic scrutiny than any comparable groupof insects (Vane-Wright, 2003). It has been the focus of numerousmolecular studies from the single-gene humble beginnings ofmolecular systematics in the 1990s to recent total evidence studiesbased on multiple genes and numerous morphological characters(see Simonsen et al., 2011 for a review). As such the recent historyof phylogenetic studies of Papilionidae is almost a mini pictureof the development of molecular and total evidence studies ofinsects.

Whereas the monophyly of Papilionidae has not been seriouslyquestioned (see Vane-Wright, 2003 for a review), the interrela-tionships between the three principal subfamilies Baroniinae, Par-nassiinae and Papilioninae have recently been the subject of somecontroversies (Fig. 8). Based purely on morphological evidence,there has never been any real doubt that Baroniinae (comprisingonly the Mexican relict species Baronia brevicornis) is the sistergroup of a clade comprising Parnassiinae and Papilioninae(Fig. 8AeB). Three convincing synapomorphies uniting the last twosubfamilies have earlier been reported:

1. Third anal vein of hind wing absent (Miller, 1987).

Fig. 7. Potential presence of forewing CuP in butterflies. AeC: Hedylidae. D. Papilionidae. A: Macrosoma napiaria with no trace of CuP. B: The structure in Macrosoma hyacinthina(black arrows), which is more likely a fold and not a vestigial vein. C: Close up of C, illustrating how the fold (arrows) does not originate on the Cu stem. D: The short, stout spuroriginating on the Cu-stem in Papilio machaon.The white arrows in B and C points to the similarly structured fold in the discal cell.


2. Cervical membrane with ventral sclerite (Miller, 1987).3. Spinasternum produced laterally at spina (de Jong et al., 1996).

The first multiple gene study of Papilionidae by Caterino et al.(2001) based on three genes (COI, COII, and Elongation Factor 1Alpha) conformed this overall relationship between the threefamilies (Fig. 8C). The first total evidence study including allsubfamilies of swallowtails (Wahlberg et al., 2005) did also to someextent support this, although the parsimony analyses tended togroup Baroniinae and Parnassiinae. Two recent, large-scale studiesfocusing on Parnassiinae, but including a number of other papilionidgenera (Nazari et al., 2007, 7 genes and 236 morphological charac-ters; Michel et al., 2008, 4 genes) both found that Baroniinae andParnassinae grouped together (Fig. 8DeE). Michel et al. (2008) didnot include morphological characters, and Nazari et al. (2007)reasonably omitted potential characters (including the last two lis-ted above) they could not double check. Simonsen et al. (2011)focused on the subfamily Papilioninae but included Baronia andfour Parnassinae genera (of 8 total) in a study that included 7 genes(same as in Nazari et al., 2007) and 94 morphological characters(based on Miller’s, 1987 dataset, but with a full re examination ofadults of all species included). In their re-examination of characters,Simonsen et al. (2011) found further evidence for swallowtailmonophyly as well as the sister group relationship between Par-nassinae and Papilioninae to the exclusion of Baronia (Fig. 8F). Themonophyly of swallowtails was found to be supported by theuniversal presence of paired, ventral sense pits on the antennae ofadults. These pits were hitherto believed to be autapomorphic forthe subordinate tribe Troidini (pipevine swallowtails) since theseminal works of Karl Jordan (see Miller, 1987 for details), but byapplying SEM it was shown that the pits are in fact present in allexamined Papilionidae. The new support for a clade comprising

Parnassinae andPapilioninae, but excluding Baronia similarly comesfrom detailed re examinations of a previously known character.Wing scales with a reticulate pattern between the longitudinalridges were thought to be a synapomorphy for a subordinate Pap-ilioninae clade comprising Teinopalpini, Papilionini and Troidini(Ghiradella, 1985; Miller, 1987). However, Simonsen et al. (2011)found these structures to be present across Parnassiinae and Papil-ioninae (albeit reduced in some subordinate groups), but absent inBaronia. Very recently the elaborate combined morphology andmolecular studyof butterflies byHeikkilä et al. (2012) found supportfor this classical arrangement of swallowtail subfamilies.

Simonsen et al. (2011) further demonstrated that despite thecentury long focus on butterfly morphology, new skeletal charac-ters are still to be discovered. Firstly, a new autapomorphy wasfound for the Troidini (despite the tribe being likely one of the beststudied groups of invertebrates). It was demonstrated that one ofthe primary types of sensilla on the antenna in the tribe (outsidethe sense pits mentioned above) are sensilla coeloconica, whichwere found in considerable numbers laterally on the segments ofthe distal half of the flagellum as well as the ‘club’. This is incontrast to the remainder of the family where the primary type ofsensilla is sensilla basiconica, with very few sensilla coeloconica oneach segment. Whereas the previous character supported analready well-supported group and thus perhaps could be said to beof limited systematic value, the next character is quite different.Within the Papilioninae tribe Leptocircini, Simonsen et al. (2011)found support from both molecular and morphological data fora controversial group comprising the old world genus Graphiumand the new world Leptocircini (Eurytides s.l.). This relationshipbasically turned classical Leptocircini phylogeny on its head, but thegroup was found to be supported by a highly specialised type ofscent scales in the male hind wing. Specialized scent scales occur

Fig. 8. Recent tribal level phylogenetic hypotheses of Papilionidae. AeB: Morphology based analyses. C, E: Molecular based analyses. D, F: Combined molecular and morphologybased analyses. CeE did not include Teinopalpini. Note the different position of Baroniinae (asterisks) in DeF. Note also that the first depicted morphological study and the latestcombined analysis are identical at this level.


widespread throughout Papilioninae, and are generally very diffi-cult to homologise across genera (see Simonsen et al., 2011), but inthis group the scales are incredibly long (>2 mm) and terminate ina very specialized “ball-like” tip unlike anything observed else-where in the family.

4. Nymphalidae

The family Nymphalidae is the most diverse within the butter-flies, both objectively in terms of species numbers (>6000 worldwide) and more subjectively in terms of morphological and bio-logical diversity (Ackery et al., 1999). Although there has not been

quite the same focus on Nymphalidae compared to Papilionidae, atleast not when measured per species, studies into phylogeny,evolutionary ecology, diversification, and zoogeography anddivergent times of Nymphalidae have to some extent been moreinstrumental in developing model systems in these fields forinsects in general (see Wahlberg andWheat, 2008; Wahlberg et al.,2009). Interestingly for a group that has received such an interestover the years, the first two major cladistic works focused onNymphalidae were molecular and not morphological. Although theclassification and evolutionary relationships of Nymphalidae havebeen frequently discussed for more than a century (see Ackery,1988; Harvey, 1991; de Jong et al., 1996 for reviews), actual


higher-level (tribal and above) phylogenetic analyses of the familybased on character sets are a relatively recent phenomenon,starting with Brower’s (2000) phylogenetic analysis of 103nymphalid genera based on 378 base pairs from the nuclear genewingless. de Jong et al. (1996) included representatives from allrecognized subfamilies but, apart from consistently placing Liby-theinae as the sister of the remaining Nymphalidae, were unable toresolve interrelationships between them. Ackery et al. (1999) notedin that respect that (p. 287) “most relationships within andbetween the various groups remain obscure”.

After Brower’s initial phylogeny, two major contributions toNymphalidae phylogenetics followed in short succession.Wahlberg et al. (2003) (Fig. 9A) published the first multiple genestudy based on 2929 base pairs from the genes Elongation Factor 1Alpha, Wingless and COI from 54 species from all 10 subfamiliesidentified by Ackery et al. (1999). The results largely supportedprevious assumptions of relationships, although the Limenitinae s.str. (white admirals) were grouped with Heliconiinae (longwingsand fritillaries), and not with the Nymphalinae (admirals and allies)e the heliconiine/limentine clade was, however, the sister of thenymphaline subfamilies. A year later Freitas and Brown (2004)published the first comprehensive morphology-based phylogenyof Nymphalidae founded on 95 species and 234 characters from alllife stages (Fig. 9B). As in the other major morphology-based workbefore them (de Jong et al., 1996), they found that a singlehypothesis was difficult to arrive at. Nevertheless, their preferredresult (a majority rule tree using all data) did have Libytheinae asthe sister group of the remaining subfamilies, followed by theDanainae s. lat. (monarchs and allies) as the sister to a large cladecomprised by the two subclades, the Nymphaloid clade and theSatyroid clade. The former comprised the Heliconiinae and Nym-phalinae (which were thus brought back together), the latter allremaining nymphalids including Limenitinae around the largesubfamily Satyrinae. Several tribes from Wahlberg et al.’s (2003)nymphaline clade were placed in the large Satyroid clade.

Fig. 9. Major subfamily level phylogenetic hypotheses of Nymphalidae. A: Molecular based aanalyses. Ithomiinae and Tellervinae in B were included in Danainae in A, C and D. BrassoSatyrinae in C and D.

Wahlberg et al.’s (2005) study of butterfly relationships was alsothe first study that combined morphological data for a broadsample (12 subfamilies) of Nymphalidae along with the three genesalso used by Wahlberg et al. (2003). Interestingly, the analyses ofmolecular data only (parsimony and Bayesian alike) failed torecover Nymphalidae as a monophyletic entity, with particularlyLibytheinae and Danainae being hard to place. Neither of themolecular-only analyses had the remaining nymphalid taxa asa clade either. The analyses of combined data did recover Nym-phalidae as monophyletic, but did not provide conclusive either tothe positions of Libytheinae and Danainae, or to the interrelation-ships of the remaining taxa.

Wahlberg and Wheat (2008) (Fig. 9C) published what may wellbe seen as the start of the phylogenomic era for not only butterflies,but for the whole Lepidoptera, analysing the phylogenetic rela-tionships between 54 nymphalid genera based on a datasetcomprised by 8114 base pairs from 11 different genetic markers (10nuclear and 1 mitochondrial). Of these, six had never been used inLepidopteraphylogenetics, twohadbeenused rarely, and three (COI,EF1-alpha and wingless) were at that point household genes inbutterfly phylogenetics. The results were generally well supported,and corroborated the five major groupings found byWahlberg et al.(2003): Libytheinae, the Danaine clade, the Satyrine clade, theHeliconiine clade (including Limenitidinae), and the Nymphalineclade. But, in contrast to both the “conventional view” andWahlberget al. (2003), Libytheinae and the Danaine clade together comprisedthe sister lineage of the rest of the family. Wahlberg et al. (2009)(Fig. 9D) combined the approaches of Freitas and Brown (2004)and Wahlberg and Wheat (2008) with a massive increase in taxonsampling in a study of 400 nymphalid genera based on 235morphological characters and up to 7733 base pairs of 10 genes (9nuclear and COI): all were sequenced for 3127 base pairs of COI, EF1-alpha and wingless; between 86 and 199 taxa had the 10 genessequenced; 238 taxa were scored for the morphological characters.The major higher-level differences between this study and the

nalyses. BeC: Molecular based analyses. D: Combined molecular and morphology basedlinae was included in Morphinae in A. Brassolinae and Morphinae were included in

Fig. 10. A: The enigmatic genus Bia (B. actorion). B: Close up of the hind wing scentorgans (thick, white arrows) and abdominal scent organs (thin black arrow) whichclearly show the genus’ affinity with Brassolini, and not Satyrinae in the old sense,although Brassolini (including Bia) are now considered part of a redefined Satyrinae(Wahlberg et al., 2009).


previousmolecular (Brower, 2000;Wahlberg et al., 2003;Wahlbergand Wheat, 2008), and combined (Wahlberg et al., 2005) studies,were that Libytheinae were well supported as the sister of theremaining Nymphalidae; and that Danainae were equally wellsupported as the sister of the rest of the family. The placement ofLibytheinaewas suggested by Ehrlich (1958) and Kristensen (1976)and appears well supported by morphology (de Jong et al., 1996;Ackery et al., 1999; Freitas and Brown, 2004), but as pointed out byWahlberg et al. (2009), it had hitherto never been conclusivelybacked bymolecular data. It is, however,worth noting that Heikkiläet al. (2012), with a more limited taxon sampling for Nymphalidae,did not recover Libytheinae as the sister group of the remainder ofthe family, but instead had LibytheinaeþDanainae in that positione taxon sampling would appear to be as crucial as charactersampling for a well supported phylogeny of Nymphalidae.

4.1. Phylogenetic position of the genus Bia

The recent history of nymphalid systematics provides excellentexamples of synergistic collaboration between molecular andmorphological approaches when combined data yield strongersupported and better resolved results at various taxonomic levels(other examples hereof include Wahlberg and Nylin, 2003;Simonsen et al., 2006; Simonsen et al., 2010; Brower et al., 2010;Penz et al., 2011). Perhaps no single problem illustrates this betterthan the systematic history of the genus Bia.

Bia (Fig. 10) is an enigmatic genus of Neotropical nymphalidbutterflies with few recognized species. While the genus indis-putably belongs to crown group Nymphalidae, its tribal andsubfamily affinities have long been contentious (reviewed by Vane-Wright and Boppré, 2005). For well over a hundred years, it hasbeen associated with crown group Satyrinae (e.g. Kirby, 1871;Weymer,1911;Miller,1968; d’Abrera,1988; Harvey,1991), althoughno convincing morphological synapomorphies support this place-ment (Vane-Wright and Boppré, 2005). In contrast to this, Freitasand Brown (2004) found that while Bia indeed was associatedwith Satyrinae s. str., it was as the sister group of a larger cladecomprising Satyrinae, Calinaginae, Morphinae and Brassolinae.Consequently they raised Bia to subfamily level (Biinae). In contrastto this, a few earlier studies (reviewed in Vane-Wright and Boppré,2005) had suggested that Bia may belong to the morphine tribeBrassolini (considered a subfamily by Freitas and Brown), but againwithout any synapomorphies being suggested. The first phyloge-netic evidence that Bia may belong to Brassolini came from Brower(2000) whose molecular phylogeny placed Bia with the other twobrassoline genera (Caligo and Opsiphanes) in both unweighted andweighted analyses. After Freitas and Brown’s (2004) somewhatambiguous placement of Bia, Vane-Wright and Boppré (2005)reviewed existing literature on Bia and carried out a detailed re-examination of the genus’s morphology in an attempt tounequivocally resolve its phylogenetic affiliations. They firstdemonstrated that Miller’s (1968) main reasons for placing Bia inSatyrinae were compromised by erroneous interpretations ofmorphology of the labial palpus and forewing venation. They thendemonstrated how the highly complex system of androconialorgans strongly supported associations with the Brassolini.Although the alar and abdominal androconial organs of Bia(Fig. 10B) bear clear overall similarities with those of other Bras-solini, the strongest evidence comes from the posterior alarandroconial organ of the hind wing, which is comprised by a seriesof long, hair-like scales arranged in palisades with conjoined scalesockets Vane-Wright and Boppré (2005, figs. 15e21). A similararrangement with hair-like scales with conjoined sockets arrangedin palisades is found in some other Brassolini genera, but areotherwise unknown from nymphalid butterflies (Vane-Wright and

Boppré, 2005; Figs. 27e31). Vane-Wright and Boppré (2005)further described hitherto overlooked serial pairs of lateralabdominal androconial organs in Bia that resemble those found inother Brassolini. Vane-Wright and Boppré (2005) further discusseda number of similarities in juvenile stages and lifestyle (unknownto Miller, 1968 and earlier authors) that also support relationshipbetween Bia and other Brassolini. As noted by Vane-Wright andBoppré (2005, p. 237): “The peculiarities of the androconialsystems reported here demonstrates that, even without theevidence available from knowledge of DNA sequences and earlystagemorphology, the clear relationship of Bia to the owl butterflies(CaligoHübner, 1819) and other Brassolini has literally been ‘staringus in the face’ for over 200 years.” The position of Bia as one of thebasal lineages in Brassolini has since been corroborated by detailedmolecular studies (Peña et al., 2006) and by the combined study ofWahlberg et al. (2009) e even if the circumscription of Satyrinae toinclude, amongst others, Morphinae presented therein means thatBia strictly speaking is back in Satyrinae.


5. Hesperiidae

Taxonomy of the Hesperioidea has long been dominated by thevoluminous work of Evans (1937e1955) who dealt with all speciesof skippers known at the time, and from all parts of the world. Hisapproach was, however, regional and although he recognizedsubfamilies and sometimes lower categories across the continents,he did not present a world-wide view of the Hesperioidea. The firstattempt to do so was made by Voss (1952), but his study wastoo limited (54 species) to have had much impact. Scott andWright (1990) did present a world-wide view, but their evidenceremained uncertain. Although Hesperiidae were included instudies on the phylogeny of the butterflies (de Jong et al., 1996;Ackery et al., 1999; Wahlberg et al., 2005), the representation ofthe family in the data bases was much too low to allow for far-reaching conclusions. Only in 2008, Warren et al. presenteda comprehensive study, based on sequences of three genes in 209species from all over the world, and one year later they had added49 morphological characters to the molecular data set and ana-lysed the combined set.

5.1. Coeliadinae vs. remaining Hesperiidae

In Evans’ (1937e1955) work there was no hierarchy among thesubfamilies of the Hesperiidae. Scott and Wright (1990) did notanalyse the Hesperiidae phylogenetically, but proposed a hierar-chical arrangement as follows: ((Pyrginae þ (Pyrrhopyginae þ

Fig. 11. Important Hesperiidae characters. AeB: Autapomorphies of the non-Coeliadinae Heextending over the eye from the base of the antenna. B: Dorsal view of the head of Prusiaautapomorphy of Hesperiinae, but also found in a few Trapezitinae. DeF: Basking posturHesperiinae (and other butterflies). EeF: Thymelicus sylvestris (E) and Udaspes folus (F) show

(Euschemoninae þ Coeliadinae))) þ (Trapezitinae þ (Hetero-pterinae þ (Hesperiinae þ Megathyminae)))). In other words,according to these authors the Coeliadinae had a rather subordinateposition. In all phylogenetic analyses, starting with de Jong et al.(1996), whether based on morphology, DNA or on both, the Coe-liadinae emerge as sister to the remaining Hesperiidae. Thisarrangement is supported by two autapomorphies for the non-coeliadine Hesperiidae: head with two pairs of chaetosemata,a character not found elsewhere in the Lepidoptera (Fig. 11A), andantennal base with an eye lash (lateral scale tuft extending over theeye) (Fig. 11B), equally not found elsewhere in the Lepidoptera, butsometimes reduced in non-coeliadine Hesperiidae. Since thesecharacters are so universal inside non-coeliadine Hesperiidae andcompletely absent outside, it would have been surprising if themolecules had taught us another lesson. Yet it is good to see thatthe molecules on their own (Warren et al., 2008) support theconclusion based on morphology.

5.2. Non-coeliadine Hesperiidae

Although the bulk of the Hesperiidae may form a well-supported monophyletic group, its subdivision is more problem-atic. Traditionally (following Evans, 1937e1955), it has beendivided into two groups, Pyrginae (inc. Pyrrhopyginae) and Tra-pezitinaeþHesperiinae (incl. Megathyminae), which, with fewexceptions, differ in food plant choice, the Pyrginae feeding ondicotyledonous plants, the other group on monocotyledons. There

speriidae. A: Dorsal view of the head of Hylephila fasciolata showing the eye lash (EL)na prusias showing the paired chaetosemata (CS). C: The position of M2 (arrow), anes. D: Pyrgus oileus displaying the flat wing position commonly found in most non-ing the peculiar wing position found in Hesperiinae and some Trapezitinae.


are, however, few morphological characters supporting this divi-sion. de Jong et al. (1996) could not find convincing evidence formonophyly of Pyrginae, let alone a sister group relationshipbetween Pyrginae and Hesperiinae. Differences are found in themale secondary sexual characters, such as a costal fold on the forewing and a tibial hair tuft on the hind leg in Pyrginae and brands orstigmata on the forewing in the other grouping, but thesecharacters are far from universal. In this case, molecules havebeen helpful. Even though only six exemplar species wereincluded, Wahlberg et al. (2005), supported the basal positionof the Coeliadinae, and found the following relationship: ((Coe-liadinaeþ ((Pyrginaeþ Pyrrhopyginae))þ (Urbanus[“Pyrginae”]þ(TrapezitinaeþHesperiinae))) (MP analysis), or (Coeliadinaeþ(Urbanus[“Pyrginae”]þ ((TrapezitinaeþHesperiinae)þ (PyrginaeþPyrrhopyginae)))) (Bayesian analysis), thus rendering the Pyrginaesensu Evans paraphyletic. The conclusions were supported andrefined in the molecular study of Warren et al. (2008) including 209species of Hesperiidae, where the monocotyledon feeders came outas a monophyletic group (to be dealt with below) and the Pyrginaeas a paraphyletic group, one group of which (viz. Eudamini, thegroup to which the genus Urbanus in Wahlberg et al., 2005 belongs)was sister to the monocot feeders (with a very poorly supportedplacement of Euschemon in between, see below).

Adding morphological characters to the molecular data base(Warren et al., 2009) did not change the overall picture in that themonocot feeders appear as sister group of a part of the traditionalPyrginae, but the subdivision of the Pyrginae changed, the“subfamily” falling into two, with the newly defined Pyrginae sisterto the monocot feeders and the Eudaminae (raised to subfamilyrank) sister to the other two combined. It is remarkable thatmorphological characters of groupings for which such evidence isscant (actually there is no morphological evidence for the mono-phyly of either Eudaminae or Pyrginae in the new sense), can sodrastically change a scheme based on molecules only. Apparently,morphological and molecular characters are each and in combi-nation too weak to lead to a robust scheme of relationships, andnew characters may easily lead to changes in the tree presented byWarren et al. (2009). This said, the latter tree is the best we haveso far.

5.3. Euschemoninae

The ever changing taxonomic position of Euschemon (Fig. 1D),a monotypic Australian genus, is illustrative of the way taxonomyhas been performed over the years, from feelings (“this species is sodifferent, it must be a separate family”) to reasoning in the earlyHennigian days (where polarity determined beforehand played animportant role), and analysing all data simultaneously in computerprograms using both parsimony and statistically based models.

Euschemon is so different from other Hesperioidea that it hasusually been considered as something different from the rest ofthe Hesperioidea. It displays a fully functional wing couplingmechanism (frenulumplus retinaculum), found in themale sexonly.Such a mechanism is widespread among other Ditrysia, but absentfrom what used to be known as Papilionoidea before new studies(Regier et al., 2009; Mutanen et al., 2010; Heikkilä et al., 2012). IfEuschemon were placed outside the Hesperioideaþ Papilionoidea(in the traditional sense), it could be thought that the wing couplingmechanism was lost only once in the evolution of the Hesper-ioideaþ Papilionoidea. Once it is seen as a member of Hesperiidae,and Hedyliidae the sister of Hesperiidae, it implies either severallosses (more so the more subordinate its position), or two reversals:one in Euschemon and another unitingHedyliidae. Onmorphologicalgrounds, Evans (1949) considered Euschemon a subordinate taxonwithin his subfamily Pyrginae. The subordinate position in

Hesperioidea was also found in the purely molecular study byWarren et al. (2008), as sister to a grouping called Eudamini byWarren et al. (2008) in the Pyrginae sensu Evans. So up to this pointmolecules and morphology came to the same conclusion.

However, when Warren et al. (2009) added 46 morphologicalcharacters to their molecular data matrix, Euschemonwas placed assister to all Hesperioidea other than Coeliadinae. For that reason thetaxon was raised to subfamily level.

The question remains whether the new position of Euschemonmakes sense in a morphological or other way. Apart from the wingcoupling mechanism, which should be considered a reversal inEuschemon, its new position agrees well with the progressivereduction of the second medial vein (M2) in the hindwing, which iswell developed in most Coeliadinae and Euschemon (as in all out-groups), but becomes weaker in the remaining Hesperioidea,particularly in the crown group Hesperiinae, where it may bevirtually absent. Therefore, apart from the wing coupling system,the position appears similarly supported by both morphologicaland molecular evidence. The new position of Euschemon alsosupports the hypothesis that the early diversification of the Hes-perioidea took place in the eastern part of the Old World tropics inthe Cretaceous (de Jong, 2007).

5.4. The monocotyledon feeders

de Jong et al. (1996) and Ackery et al. (1999) could not findmorphological evidence for keeping Megathyminae apart fromHesperiinae. The separation was only based on autapomorphiccharacters of Megathyminae directly related to the boring habit ofthe larva: larval and adult head relatively small and larva withouta “neck” (a narrow part of the body directly behind the headfacilitating movement when living between leaves spun together).The combined morphological and molecular study by Wahlberget al. (2005) did not include Megathymus, but two species of thesupposed subfamily were included in the molecular study byWarren et al. (2008). They both ended up internal to the Hesperi-inae and the same was the case when morphological characterswere added (Warren et al., 2009), so in this case morphologicalcharacters and molecular characters supported each other indowngrading the Megathymus and allies.

Apart from their food plant choice there is little to connect theHeteropterinae, Trapezitinae and Hesperiinae but molecular char-acters; only the presence of brands or stigmata of androconial scaleson the forewing of the male could be considered morphologicalsupport, but these are absent in many species, and completely inHeteropterinae, thus of dubious value. Scott and Wright (1990)mentioned a larval character (seta D1 on dorsum of first instarabdominal segment 10 long), but they did not indicate inwhich, andin how many, species they had found it, and larval characters inHesperiidae in general are incompletely known. Nevertheless,Warren et al. (2008) found strong support (BS¼ 9) for this grouping,a conclusion that was not changed when morphological data wereadded to the molecular data matrix (Warren et al., 2009).

While Evans (1949) included the Heteropterinae in the Hes-periinae, he kept the Trapezitinae separate because of a peculiarautapomorphy in the venation of the hind wing (discocellular veinsform a line directed to the apex of the hind wing; in other Hes-periidaethe line is vertical or directed somewhat towards the wingbase). As a consequence Heteropterinae and Hesperiinae wereunited on the basis of a symplesiomorphy (“H cell normal”, Evans,1949: 2). Not surprisingly, Evans’ subdivision was not supported bymolecular characters. Warren et al. (2008) found some, but ratherweak support (Bremer Support¼ 2) for a sister group relationshipof Trapezitinae and Hesperiinae. This relationship was not changedby adding morphological characters (Warren et al., 2009). For this


grouping and the exclusion of Heteropterinae, morphological orbiological evidence is not very strong in the sense of universality:androconial patches in the form of brands or stigmata on the forewing of the male occur exclusively in Trapezitinae and Hesperiinae,but they are absent in many species. The peculiar basking posture,with hind wings held flat and fore wings only partly open, isexclusively found in the two subfamilies (Fig. 11EeF). However,while it is almost universal in Hesperiinae, it occurs in only four ofthe eleven genera in Trapezitinae.

The monophyly of both Heteropterinae and Trapezitinae is wellsupported by molecular characters, with BS¼ 14 and BS¼ 19,respectively (Warren et al., 2008). In view of the limited morpho-logical evidence (only one character for Trapezitinae, see above;non-unique characters like long, porrect, hairy palps and a longhind wing cell for Heteropterinae), such high support is surprising.The Hesperiinae, containing the bulk of the species in the family,are morphologically only characterized by a shift of the base of veinM2 in the forewing towards M3 (Fig. 11C), a character not alwaysunambiguous, and sometimes found in Trapezitinae as well. Itsmonophyly finds support in DNA (BS¼ 5).

Since there is no purely morphological study on the phylogenyof the Hesperioidea comparable to Warren et al. (2008) available, itis not easy to judge how far molecular characters have yieldedphylogenetic information that could not be found bymorphologicalcharacters. As matters stand, for the higher categories of the Hes-periidae molecules have either supported previous ideas of rela-tionships and monophyly, or have refined relationships wheremorphological characters were insufficient.

6. Concluding remarks

Butterfly systematics has come a long way since Linnaeus (1758)presented the first subdivision. It is remarkable that the subdivisioninto what we now call families has changed little since Latreille(1805), and the scheme given by Bates (1864), almost 150 yearsold now, looks remarkably modern (see de Jong et al., 1996). Sinceup toWahlberg et al. (2005) most research on family level butterflysystematics was morphological and since the inclusion of molec-ular characters did not suggest radical changes for the time being,morphological characters were apparently a good guide to thephylogeny, at least at the family level. Themore remarkablewas thefinding of Regier et al. (2009), Mutanen et al. (2010) and Heikkiläet al. (2012) that there was molecular evidence for a radicalchange in our idea of butterfly relationships, Hesperiidae andHedylidae being sister groups, and Papilionoidea being non-monophyletic. Such a scheme was foreshadowed in the moleculartree based on Bayesian analysis in Wahlberg et al. (2005), but, asMutanen et al. (2010) put it, “this result was obscured bymorphological evidence”. Such a hypothesis is controversial as wellas intriguing. However, we should not blame morphology for thelate discovery, but take a new look at the morphology as well as atthe molecules, since also the molecules can tell an incorrect story.The conflicting results from the large-scale, molecular-only studiesof Regier et al. (2009) and Mutanen et al. (2010) clearly demon-strate that even “genomic size” datasets are not without conflicts.Furthermore, the evidence from immature life stages in favor ofa monophyletic Rhopalocera (including Hedylidae) found byHeikkilä et al. (2012) clearly demonstrates how careful studies oflittle used character sources (in this case the under-utilizedimmature life stages) can provide new evidence in support of oneof several conflicting hypotheses.

This also seems tobe the case (albeit based on adultmorphology)in the subfamily taxonomy of the Papilionidae, a very simple one,withBaroniinae sister to theParnasiinaeþ Papilioninae, as generallyfound in morphological and early molecular studies, or Baroniinae

sister to Parnasiinae as in two recent studies (Nazari et al., 2007;Michel et al., 2008), based solely (the latter) or partly (the former) onmolecular characters. Promptedby this result, Simonsen et al. (2011)went back to the morphological characters and found furtherevidence for the traditional scheme.

The synergistic effect of combining morphological and molec-ular data is well illustrated by the taxonomy of two large families,Nymphalidae and Hesperiidae. The latter case is particularly illu-minating as the authors (Warren et al., 2008, 2009) analysed(almost) the same set of taxa with molecules only and withmorphological characters added. This addition of morphologicalcharacters had a profound effect on the position of some taxa,showing that molecules on their own not necessarily lead toa robust phylogeny. The addition of new characters, be theymorphological, molecular or otherwise may be indispensable fora robust result. Of course, the synergistic effect works both ways. Itis worth remembering that when morphology was analysed alone,Heikkilä et al. (2012) did not recover the family level relationshipsdiscussed here. Only the addition of molecules enabled them toreach a well-supported result.

Finally, it should not be forgotten that drawing up a phyloge-netic tree should not be a goal in itself, but ameans towards a betterunderstanding of evolution. Molecules may help us to get a betterinsight into the evolution of particular characters (such as thedisappearance of vein M2 in the hind wing in Hesperioidea asdescribed above) or show us that a supposed synapomorphyactually is a parallelism, as the dwindling anepisternum in Papil-ionidae is, if its new position is further supported. A robustphylogeny is also helpful in detecting underlying synapomorphies,as seem to abound in Hesperioidea (e.g. male secondary sexualcharacters) or reversals, which may be called an extreme form ofunderlying synapomorphies. In a group of taxa with an underlyingsynapomorphy, the taxon that does not show it is like a hard disk onwhich information is present, but not accessible because of anelectronic block. In due course the molecules may tell us what themolecular block is for an underlying synapomorphy, but there isstill a long way to go. For the time being, we can only drawhypotheses about underlying synapomorphies from a robustphylogeny, not the other way around.

Acknowledgments

Wewish to thank Niels P. Kristensen, Natural HistoryMuseum ofDenmark, and Rolf G. Beutel, Institut für spezielle Zoologie undEvolutionsbiologie, FSU Jena for the invitation to write this reviewarticle. Marie Djernæs and Ian J. Kitching, Natural History Museum,London are thanked for discussions on methodology, morphologyand character evolution. MH & LK were funded by Academy ofFinland through grant awarded to LK (project 1110906).

References

Ackery, P.R., 1988. Hostplants and classification: a review of nymphalid butterflies.Biological Journal of the Linnean Society 33, 95e203.

Ackery, P.R., de Jong, R., Vane-Wright, R.I., 1999. The butterflies: Hedyloidea, Hes-perioidea and Papilionoidea. In: Kristensen, N.P. (Ed.), Lepidoptera, Moths andButterflies. Evolution, Systematics, and Biogeography. Handbook of Zoology,vol. 1. Walter de Gruyter, Berlin, pp. 399e436. 4(35).

Bates, H.W., 1864. Contributions to an insect fauna of the Amazon valley. Lepi-doptera e Nymphalinae. Journal of Entomology 2, 175e213.

Brock, J.P., 1971. A contribution towards an understanding of the morphology andphylogeny of the ditrysian Lepidoptera. Journal of Natural History 5, 29e102.

Brower, A.V.Z., 2000. Phylogenetic relationships among the Nymphalidae (Lepi-doptera), inferred from partial sequences of the wingless gene. Proceedings ofthe Royal Society of London Series B, Biological Sciences 267, 1201e1211.

Brower, A.V.Z., Wahlberg, N., Ogawa, J.R., Boppré, M., Vane-Wright, R.I., 2010.Phylogenetic relationships among genera of danaine butterflies (Lepidoptera:Nymphalidae) as implied by morphology and DNA sequences. Systematics andBiodiversity 8, 75e89.


Caterino, M.S., Reed, R.D., Kuo, M.M., Sperling, F.A.H., 2001. A partitioned likelihoodanalysis of swallowtail butterfly phylogeny (Lepidoptera: Papilionidae).Systematic Biology 50, 106e127.

d’Abrera, B., 1988. Butterflies of the Neotropical Region, Part 5: Nymphalidae(conc.), Satyridae. Hill House Publishers, Melbourne, 190 pp.

de Jong, R., 2007. Estimating time and space in the evolution of the Lepidoptera.Tijdschrift voor Entomologie 150, 319e346.

de Jong, R., Vane-Wright, R.I., Ackery, P.R., 1996. The higher classification of butterflies(Lepidoptera): problems and prospects. Entomologica scandinavica 27, 65e101.

Ehnbom, K., 1948. Studies on the central and sympathetic nervous system and somesense organs in the head of neuopteroid insects. Opuscula entomologica(Suppl. 8), 1e162.

Ehrlich, P.R., 1958. The comparative morphology, phylogeny, and higher classifica-tion of butterflies (Lepidoptera: Papilionidae). University of Kansas ScienceBulletin 34, 305e370.

Ehrlich, P.R., 1960. The integumental anatomy of the silver-spotted skipper, Epar-gyreus clarus Cramer (Lepidoptera: Hesperiidae). Microentomology 24, 1e23.

Ehrlich, P.R., 1961. Comparative morphology of the male reproductive system of thebutterflies (Lepidoptera: Papilionoidea). 1. Some nearctic species. Micro-entomology 24, 135e166.

Ehrlich, P.R., Davidson, S.E.,1961. The internal anatomyof themonarch butterfly, Danausplexippus L. (Lepidoptera: Nymphalidea). Microentomology 24, 85e133.

Ehrlich, P.R., Ehrlich, A.H., 1962. The head musculature of the butterflies (Lepi-doptera: Papilionoidea). Microentomology 25, 1e89.

Ehrlich, P.R., Ehrlich, A.H., 1963. The thoracic and basal abdominal musculature ofthe butterflies (Lepidoptera: Papilionoidea). Microentomology 25, 91e126.

Ehrlich, P.R., Ehrlich, A.H., 1967. The phenetic relationships of the butterflies. I. Adulttaxonomy and the nonspecificity hypothesis. Systematic Zoology 16, 301e317.

Evans, W.H., 1937. A Catalogue of the African Hesperiidae Indicating the Classifi-cation and Nomenclature Adopted in the British Museum (Natural History).British Museum (Natural History), London, xii þ 212, pp. 30.

Evans, W.H., 1949. A Catalogue of the Hesperiidae from Europe, Asia and Australia inthe British Museum (Natural History). British Museum (Natural History), Lon-don, xix þ 512, pp. 52.

Evans, W.H., 1951e55. A Catalogue of the American Hesperiidae Indicating theClassification and Nomenclature Adopted in the British Museum (NaturalHistory). Part I. Introduction and Group A. Pyrrhpyginae x þ 92 pp; Part II.(Groups B, C, D) Pyrginae. Section 1 v þ 178 pp; Part III. (Groups E, F. G) Pyr-ginae. Section 2 c þ 246 pp; Part IV (Groups H-P) Hesperiinae and Mega-thyminae. British Museum (Natural History), London, v þ499 pp.

Freitas, A.V.L., Brown, K.S., 2004. Phylogeny of the Nymphalidae (Lepidoptera).Systematic Biology 53, 363e383.

Ghiradella, H., 1985. Structure and development of iridescent lepidopteran scales:the Papilionidae as a showcase family. Annals of the Entomological Society ofAmerica 78, 252e264.

Harvey, D., 1991. Appendix B: higher classification of the Nymphalidae. In:Nijhout, H.F. (Ed.), The Development and Evolution of Butterfly Wing Patterns.Smithsonian Institution Press, Washington, DC, pp. 255e273.

Heikkilä, M., Kaila, L., Mutanen, M., Peña, C., Wahlberg, N., 2012. Cretaceous originand repeated tertiary diversification of the redefined butterflies. Proceedings ofthe Royal Society B, 279 (1731), 1093e1099.

Hessel, J.H.,1966. Apreliminary comparative anatomical studyof themesothoracic aortaof the Lepidoptera. Annals of the Entomological Society of America 59, 1217e1227.

Hessel, J.H., 1969. The comparative morphology of the dorsal vessel and accessorystructures of the Lepidoptera and its phylogenetic implications. Annals of theEntomological Society of America 62, 353e370.

Kirby, W.F., 1871. A Synoptic Catalogue of the Diurnal Lepidotera, Viii þ 690 pp,London.

Kristensen, N.P., 1976. Remarks on the family-level phylogeny of butterflies,(Insecta, Lepidoptera, Rhopalocera). Zeitschrift für Zoologische Systematik undEvolutionsforschung 14, 25e33.

Kudrna, O. (Ed.), 1990. Butterflies of Europe 2. AULA-Verlag, Wiesbaden, p. 557.Latreille, P.A., 1805. Histoire naturelle des Crustacés et Insectes. 14. 432 pp., Paris.Linnaeus, C., 1758. Systemae Naturae (Edn. 10). 823 pp., Stockholm.Maddison, W.P., Maddison, D.R., 2011. Mesquite: a modular system for evolutionary

analysis. Version 2.75. http://mesquiteproject.org.Michel, F., Rebourg, C., Cosson, E., Decimon, H., 2008. Molecular phylogeny of Parnas-

siinae butterflies (Lepidoptera: Papilionidae) based on the sequences of four mito-chondrial DNA segments. Annales de la Société Entomologique de France 44, 1e36.

Miller, L.D., 1968. The higher classification, phylogeny and zoogeography of the Satyr-idae (Lepidoptera). Memoirs of the American Entomological Society 24, 1e174.

Miller, J.S., 1987. Phylogenetic studies in the Papilioninae (Lepidoptera: Papil-ionidae). Bulletin of the American Museum of Natural History 186, 365e512.

Minet, J., 1988. The major ditrysian lineages and their relationships (Lepidoptera).Proceedings of the International Congress of Entomology 18, 79 (Abstracts andauthor index).

Minet, J., 1991. Tentative reconstruction of the ditrysian phylogeny (Lepidoptera:Glossata). Entomologica scandinavica 22, 69e95.

Mutanen, M., Wahlberg, N., Kaila, L., 2010. Comprehensive gene and taxon coverageelucidates radiation patterns in moths and butterflies. Proceedings of the RoyalSociety Series B Biological Sciences 277, 2839e2848.

Nazari, V., Zakharov, E.V., Sperling, F.A.H., 2007. Phylogeny, historical biogeog-raphy, and taxonomic ranking of Parnassiinae (Lepidoptera, Papilionidae)based on morphology and seven genes. Molecular Phylogenetics Evolution 42,131e156.

van Nieukerken, E.J., Kaila, L., Kitching, I.J., Kristensen, N.P., Lees, D.C., Minet, J., Mitter, C.,Mutanen, M., Regier, J.C., Simonsen, T.J., Wahlberg, N., Yen, S.-H., Zahiri, R.,Adamski, D., Baixeras, J., Bartsch, D., Bengtsson, B.Å., Brown, J.W., Bucheli, S.R.,Davis, D.R., De Prins, J., De Prins, W., Epstein, M.E., Gentili-Poole, P., Gielis, C.,Hättenschwiler, P., Hausmann, A., Holloway, J.D., Kallies, A., Karsholt, O.,Kawahara, A.Y., Koster (Sjaak), J.C., Kozlov,M.V., Lafontaine, J.D., Lamas, G., Landry, J.-F., Lee, S., Nuss, M., Park, K.-T., Penz, C., Rota, J., Schmidt, B.C., Schintlmeister, A.,Sohn, J.-C., Solis, M.A., Tarmann, G.M., Warren, A.D., Weller, S., Yakovlev, R.V.,Zolotuhin, V.V., Zwick, A., 2011. Order Lepidoptera Linnaeus, 1758. In: Zhang, Z.-Q.(Ed.), Animal Biodiversity: An Outline of Higher Classification and Survey of Taxo-nomic Richness. Zootaxa, vol. 3148, pp. 212e221.

Peña, C., Wahlberg, N., Weingartner, E., Kondandaramaiah, U., Nylin, S.,Freitas, A.V.L., Brower, A.V.Z., 2006. Higher level phylogeny of Satyrinaebutterflies (Lepidoptera: Nymphalidae) based on DNA sequence data. MolecularPhylogenetics and Evolution 40, 29e49.

Penz, C.M., Mohammadi, N., Wahlberg, N., 2011. The Neotropical butterfly genusBlepolenis: wing pattern elements, phylogeny, and Pleistocene diversification(Lepidoptera, Nymphalidae). Zootaxa 2897, 1e17.

Regier, J.C., Zwick, A., Cummings, M.P., Kawahara, A.Y., Cho, S., Weller, S., Roe, A.,Baixeras, J., Brown, J.W., Parr, C.,Davis, D.R., Epstein,M.,Hallwachs,W.,Hausmann,A.,Janzen,D.H., Kitching, I.J., Solis,M.A., Yen, S.-H., Bazinet,A.L.,Mitter, C., 2009. Towardsreconstructing the evolution of advanced moths and butterflies (Lepidoptera:Ditrysia): an initial molecular study. BMC Evolutionary Biology 9, 280.

Scoble, M.J., 1986. The structure and affinities of the Hedyloidea: a new concept ofbutterflies. Bulletin of the British Museum of Natural History (Ent) 53, 251e286.

Scoble, M.J., Aiello, A., 1990. Moth-like butterflies (Hedylidae: Lepidoptera):a summary, with comments on the egg. Journal of Natural History 24, 159e164.

Scott, J.A., 1985. The phylogeny of butterflies (Papilionoidea and Hesperioidea).Journal of Research on the Lepidoptera 23, 241e281.

Scott, J.A., Wright, D.M., 1990. Butterfly phylogeny and fossils. In: Kudrna (Ed.),Butterflies of Europe. 2. AULA Verlag, Wiesbaden, pp. 152e208.

Simonsen, T.J., Wahlberg, N., Brower, A.W.Z., de Jong, R., 2006. Molecules, morphologyand Fritillaries: a combined approach towards the phylogeny of Argynnini (Lepi-doptera: Nymphalidae). Insect Systematics and Evolution 37, 405e418.

Simonsen, T.J., Wahlberg, N., Warren, A.D., Sperling, F.A.H., 2010. The evolutionaryhistory of Boloria (Lepidoptera: Nymphalidae): phylogeny, zoogeography andlarval foodplant relationships. Systematics and Biodiversity 8, 513e529.

Simonsen, T.J., Zakharov, E.V., Djernaes, M., Cotton, A., Vane-Wright, R.I.,Sperling, F.A.H., 2011. Phylogeny, host plant associations and divergence time ofPapilioninae (Lepidoptera: Papilionidae) inferred from morphology and sevengenes with special focus on the enigmatic genera Teinopalpus and Meandrusa.Cladistics 27, 113e137.

Vane-Wright, R.I., 2003. Evidence and identity in butterfly systematics. In:Boggs, C.L., Watt, W.B., Ehrlich, P.R. (Eds.), Butterflies: Ecology and EvolutionTaking Flight. University of Chicago Press, Chicago, pp. 477e513.

Vane-Wright,R.I., Boppré,M.,2005.AdultmorphologyandthehigherclassificationofBiaHübner (Lepidoptera: Nymphalidae). Bonner Zoologische Beiträge 53, 235e254.

Voss, E.G., 1952. On the classification of the Hesperiidae. Annals of the Entomo-logical Society of America 45, 246e258.

Wahlberg, N., Nylin, S., 2003. Morphology versus molecules: resolution of the posi-tions of Nymphalis, Polygonia and related genera (Lepidoptera: Nymphalidae).Cladistics 19, 213e223.

Wahlberg, N., Weingartner, E., Nylin, S., 2003. Towards a better understanding ofthe higher systematics of Nymphalidae (Lepidoptera: Papilionoidea). MolecularPhylogenetics and Evolution 28, 473e484.

Wahlberg, N., Braby, M.F., Brower, A.V.Z., de Jong, R., Lee, M.-M., Nylin, S., Pierce, N.,Sperling, F.A.H., Vila, R., Warren, A.D., Zakharov, E.V., 2005. Synergistic effects ofcombining morphological and molecular data in resolving the phylogeny ofbutterflies and skippers. Proceedings of the Royal Society of London Series BBiological Sciences 272, 1577e1586.

Wahlberg, N., Wheat, C.W., 2008. Genomic outposts serve the phylogenomicpioneers: designing novel nuclear markers for genomic DNA extractions ofLepidoptera. Systematic Biology 57, 231e242.

Wahlberg, N., Leneveu, J., Kodandaramaiah, U., Peña, C., Nylin, S., Freitas, A.V.L.,Brower, A.V.Z., 2009. Nymphalid butterflies diversify following near demise atthe Cretaceous/Tertiary boundary. Proceedings of the Royal Society Series BBiological Sciences 276, 4295e4302.

Warren, A.D., Ogawa, J.R., Brower, A.V.Z., 2008. Phylogenetic relationships ofsubfamilies and circumscriptions of tribes in the family Hesperiidae (Lepidop-tera: Hesperioidea). Cladistics 24, 642e676.

Warren, A.D., Ogawa, J.R., Brower, A.V.Z., 2009. Revised classification of the familyHesperiidae (Lepidoptera: Hesperioidea) based on combined molecular andmorphological data. Systematic Entomology 34, 467e523.

Weintraub, J.D., Miller, J.S., 1987. The structure and affinities of the Hedyloidea:a new concept of butterflies. Cladistics 3, 299e304.

Weymer, G., 1911. 4. Family Satyridae. In: Seitz, A. (Ed.), Die Grosshemetterlinge derErde 5. A. Kernen, Stuttgart, pp. 173e283. 19 pls.

arthropod structure & developmentigarzon.weebly.com/.../simonsen_etal_2012_review.pdf · 310 t.j....

Documents