Systematic Entomology (2003) 28, 279–322 Cladistic analysis of the Neotropical butterfly genus Adelpha (Lepidoptera: Nymphalidae), with comments on the subtribal classification of Limenitidini KEITH R. WILLMOTT Department of Entomology, The Natural History Museum, London, U.K. Abstract. A two-stage cladistic analysis of 114 characters from adult and imma- ture stage morphology provided phylogenetic hypotheses for the diverse Neotrop- ical nymphalid butterfly genus Adelpha Hu¨ bner. Higher-level cladistic relationships were inferred for thirty Adelpha species and twenty other species of Limenitidini, confirming the monophyly of Adelpha as currently conceived and indicating several montane Asian species as potential sister taxa for the genus. Cladistic relationships between all eighty-five Adelpha species were then inferred using three outgroup combinations. Basal and terminal nodes were reasonably resolved and supported, but a low proportion of non-wing pattern characters resulted in weak resolution and support in the middle of the tree. The most basal members of Adelpha feed on the temperate or montane plant family Capri- foliaceae, suggesting that a switch from this family early in the evolutionary history was important in subsequent diversification into tropical lowland habitats. The cladograms confirm suspicions of earlier authors that dorsal mimetic wing patterns have convergently evolved a number of times in Adelpha. The subtribal classification of Limenitidini is discussed and both Lebadea (from Parthenina) and Neptina are transferred to Limenitidina, whereas Cymothoe, Bhagadatta and Pseudoneptis (all formerly Limenitidina) are regarded as incertae sedis. Introduction Butterflies provide a rich source of material for many topics western U.S.A. to Uruguay (Willmott, 2003). All species in evolutionary biology, and the development of computer- occur in forest habitats, from sea level to over 3000 m. based cladistic analysis in recent decades has provided a Diversity peaks at the base of the eastern Andes, where much firmer foundation for such studies. Phylogenetic local faunas may include up to thirty-nine species work on butterflies has illuminated subjects as diverse as (Willmott, 2003), certainly the highest community species ecological modes of speciation (e.g. Turner, 1976), evolu- richness for any Neotropical nymphalid genus. The dorsal tionary shifts in host-plant and chemical ecology (e.g. wing surface is typically arrayed with very simple but bright Brown et al., 1991; Wahlberg, 2001), chemical communica- colours, and although a significant number of species have tion (e.g. Schulz et al., 1993), and Amazonian (e.g. Brower, been described only recently (DeVries & Chaco´ n, 1982; 1996; Hall & Harvey, 2002), Gondwanan (e.g. Parsons, Willmott & Hall, 1995, 1999; Austin & Jasinski, 1999; 1996) and Andean (e.g. Willmott et al., 2001) biogeography. Willmott, 2003), many other species are common and therefore Adelpha is the only Neotropical member of tribe Lime- highly conspicuous forest butterflies. The immature stages nitidini, and is one of the largest Neotropical nymphalid are diverse in morphology, behaviour and food-plant genera, with eighty-five species distributed from far north- specialization (Aiello, 1984; Willmott, 2003), and the genus thus offers great promise for evolutionary study. However, such study has been hindered by a chaotic nomenclature Correspondence: Keith R. Willmott, Department of Entomo- and poor understanding of species limits and identification, logy, The Natural History Museum, Cromwell Road, London SW7 now addressed in a revision of the genus (Willmott, 2003), 5BD, U.K. E-mail: [email protected] and the absence of any phylogenetic hypothesis to date. # 2003 The Royal Entomological Society 279 280 Keith R. Willmott Perhaps the first attempt to define relationships between lysis including fewer outgroup taxa, selected following the Adelpha species was that of Godman & Salvin (1884), who results of the first analysis, all Adelpha species, and add- arranged the known Central American species in a dichot- itional characters. This permitted the inclusion of additional omous key. Although apparently the first to have studied characters in the species-level analysis that otherwise vary the male genitalia, they still relied mainly on characters of too much to allow coding, and reduced the likelihood of the eyes and dorsal wing pattern. Their arrangement was mistaken homology assessment of characters in distantly presumably designed to aid identification rather than to related taxa. indicate evolutionary relationships, but it had a great influ- ence on all subsequent authors and the curation of most museum collections. Study taxa and outgroup choice Fruhstorfer (1915), in the first major review of the entire genus, largely followed Godman & Salvin’s (1884) order of Higher-level analysis. The selection of which species (out- species, and also made some use of male genitalia and side Adelpha) to include in the higher-level analysis (and forewing venation. Nevertheless, greatest weight was still therefore as potential outgroup species for the lower level) placed on characters of the dorsal wing pattern. was the most complex aspect of this study. Adelpha is a Moss (1933) first drew attention to the possibility that the member of Limenitidini, a tribe currently placed in the dorsal wing pattern might be a poor indicator of phylogeny, para- or polyphyletic subfamily Biblidinae (Brower, 2000). with his study of the immature stages of eleven Brazilian Limenitidini is therefore equally likely to be the sister group species. Aiello (1984) expanded on this theme and collated to any of several other nymphalid subfamilies (Harvey, all published information on Adelpha life histories to 1991; Brower, 2000). Limenitidini, as treated here, has attempt to define more ‘natural’ species groups. She studied been recognized by all modern authors (Chermock, 1950; eighteen species and recognized seven species groups, pro- Eliot, 1978; Harvey, 1991), but the principal evidence for its viding a key to their identification, but did not intend this to monophyly still remains the structure of the eggs: in all represent cladistic relationships. The remaining sixty-seven known species these are composed of polygonal, sunken species in the genus were omitted from the classification. facets, and in almost all known species there are short, Otero & Aiello (1996) subsequently suggested that a new, hairlike projections at the vertices (Harvey, 1991; Igarashi eighth, species group might be required for A. alala, and five & Fukuda, 1997, 2000). These projections may be chorionic additional species of which the life histories were unknown sculpturing or aeropylar tubes, as suggested by Amiet were also listed as possible members. (2000a). Exceptions include several species of Pseudacraea The continually increasing body of knowledge on imma- Westwood (Amiet, 2000b), Pseudoneptis bugandensis ture stage morphology suggests deep flaws in the higher- Stoneham and at least one species of Neptis Fabricius and level taxonomic arrangements of earlier authors, however Catuna Kirby (Amiet, 2002), which lack these fine projec- ill-defined, and Aiello (1984) and Otero & Aiello (1996) tions. Chermock (1950) and Harvey (1991) defined the tribe suggested that the genus might even prove to be paraphy- on the basis of a single character, the preservation of the letic with respect to certain Asian limenitidines. In addition, first anal vein (1A) as a short spur at the base of the species-level taxonomic study reveals substantial racial vari- forewing cubital vein. However, this vein is not present in ation in the dorsal wing pattern (Willmott, 2003), suggest- Neptis and its relatives (Chermock, 1950), and also occurs ing that a classification based almost exclusively on such a in certain members of Heliconiinae, as noted by Michener character set is unlikely to reflect accurately phylogenetic (1942) and Brown & Heineman (1972), including the genera patterns. Indeed, a number of conspecific taxa were impli- Philaethria Billberg, Dryas Hu¨ bner and Dryadula Michener citly included in different ‘species groups’ by Fruhstorfer (C. Penz, personal communication). Indeed, it is possible (1915) and other authors (see Willmott, 2003). This study that Limenitidini are closely related to Heliconiinae therefore had several goals: to test the value of different (Brower, 2000). One behavioural trait characteristic of the character sources in providing phylogenetic information, tribe is the habit of resting with the wings open, and the both within Adelpha and within Limenitidini; to test the distinctive, gliding flight with the wings pointed downwards monophyly of Adelpha; and to provide the first comprehen- (Eliot, 1978; personal observation). Finally, recent trees sive phylogenetic hypothesis for the genus. generated by cladistic analysis of equally weighted molecu- lar characters also supported monophyly of the tribe, although the most parsimonious tree (MPT) after successive Methods approximations character weighting (SACW) did not (Brower, 2000). Nevertheless, I conclude that monophyly Analytical approach of Limenitidini is sufficiently well supported to assume that the closest relatives of Adelpha lie within the tribe. I there- The two main goals of this study, testing the monophyly fore examined the wing patterns of the majority of species in of Adelpha and producing a hypothesis of species relation- all Limenitidini genera in museum collections (Allyn ships, were