Phylogeny of the Neotropical moth tribe Josiini (Notodontidae: Dioptinae): a hidden case of Müllerian mimicry

Zoological Journal of the Linnean Society (1996), 118: 1–45. With 23 figures

Phylogeny of the Neotropical moth tribe Josiini (Notodontidae: Dioptinae): a hidden case of Mullerian ¨ mimicry JAMES S. MILLER American Museum of Natural History, Department of Entomology, Central Park West at 79th Street, New York, NY, 10024, U.S.A.

Received December 1994, accepted for publication July 1995

The Neotropical moth tribe Josiini (Notodontidae: Dioptinae) contains over 100 described species in 11 genera. All are diurnal, with brightly-coloured, presumably aposematic wing patterns. Larval hostplants are exclusively in the genus Passiflora (Passifloraceae) except for two new records, reported here, from Turnera (Turneraceae). A comparative morphological study of 26 representative josiine species yielded 86 characters from adults, larvae and pupae, all of which are figured and discussed. Phylogenetic analysis of these data produced a single most-parsimonious cladogram. According to the phylogenetic results: (1) monophyly of the Josiini is strongly supported; (2) the currently accepted generic classification is in disarray; (3) morphological character variation is extensive, and adult traits reflect phylogeny more effectively than do those of immature stages; (4) wing pattern types have undergone convergent evolution. A rare phenotype, longitudinal wing stripes, appears in two widely divergent clades, suggesting the evolution of M¨ullerian mimicry within the Josiini. ©1996 The Linnean Society of London

ADDITIONAL KEY WORDS: — Lepidoptera – morphology – wing pattern evolution – cladistics. CONTENTS Introduction . . . . . . . . . . Methods . . . . . . . . . . . Species sample . . . . . . . . Hostplants . . . . . . . . . Morphology . . . . . . . . Cladistic analyses . . . . . . . Results . . . . . . . . . . . . Cladograms . . . . . . . . . Morphology . . . . . . . . Discussion . . . . . . . . . . . Comparison of characters from adults Wing pattern evolution . . . . . Classification of the Josiini . . . . Acknowledgements . . . . . . . . References . . . . . . . . . . . Appendix I . . . . . . . . . .

0024–4082/96/090001 + 45 $18.00/0

. . . . . . . . . . and . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . immatures . . . . . . . . . . . . . . . . . . . .

1

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . .

2 3 3 3 8 8 9 9 9 35 35 37 40 41 41 45

©1996 The Linnean Society of London

2

J. S. MILLER INTRODUCTION

Patterns of diversity among tropical Lepidoptera engaged the curiosity of nineteenth century biologists, and they continue to provide a wealth of natural history problems today. Bates’ (1862) observations on wing pattern resemblance and taxonomic relationships among Amazonian butterflies not only led him to question Creation, but also kindled what has become one of the most intensively explored fields in evolutionary biology — the study of mimicry. Recent methodological advances have opened two new channels of inquiry into patterns and mechanisms of lepidopteran evolution. These are having profound, sometimes provocative, implications for mimicry theory. First, developmental researchers are now identifying the mechanisms underlying wing pattern expression in butterflies (Nijhout, 1990, 1991; Nijhout & Wray, 1986, 1988). The studies reveal that phenotype, even in species with highly intricate patterns, is controlled by a limited set of organizing centres on the wing. The evolution of mimetic patterns may occur through relatively small changes in these modular elements (Nijhout, Wray & Gilbert, 1990; Nijhout, 1994a). Concurrently, advances in Drosophila developmental genetics are providing new insights into the genetics of wing pattern formation in butterflies (Nijhout, 1994b). Carroll et al. (1994) discovered that elements of the wing pattern in the buckeye butterfly, Precise coenia (Nymphalidae), are governed by the same genes that control wing expression in Drosophila, suggesting highly conserved developmental mechanisms. However, in the buckeye most of these gene homologues show slightly different patterns of expression, and some genes exhibit functions unique to butterflies, such as in the formation of eyespots. The second research field providing new perspectives on mimicry results from a growing interest in the application of phylogenetic methods to studies in historical ecology (e.g. Miles & Dunham, 1993; Miller & Wenzel, 1995). Within this context, certain famous cases of mimicry are being reexamined. Although lepidopteran wing patterns have served as the primary character system for the group’s taxonomy, these recent studies are for the first time offering precise hypotheses with respect to the evolutionary transformation of wing pattern traits. The results conflict with longstanding theories. For example, the remarkable mimicry between geographical race pairs of Heliconius erato and H. melpomene in South America has traditionally been explained as a case of coevolution between the two butterfly species (Sheppard et al., 1985). Brower (1994a, 1996), however, found little evidence to support a strict coevolutionary relationship between H. erato and H. melpomene mimetic races. Instead, cladograms from DNA sequences suggest a highly complex evolutionary history with considerable convergence in wing pattern phenotypes, even within species (Brower, 1994b). Vane-Wright & Smith (1991) used phylogenetic analysis of species in the African Papilio phorcas group to show that the sequence of wing pattern evolution in mimetic female morphs of Papilio dardanus should be completely reversed from the previously accepted transformation hypothesis. Both of these studies highlight the critical need for studying mimicry from the standpoint of phylogeny. In this paper, cladograms based on morphological characters set the historical framework for studying wing pattern evolution in a tribe of Neotropical moths, the Josiini (Notodontidae: Dioptinae). Although the biology and taxonomy of the Josiini are poorly known compared to most butterfly groups, these moths provide a model

PHYLOGENY OF THE JOSIINI

3

system for studying mimicry. The Josiini is a fairly large clade; as currently defined the tribe includes 103 species in 11 genera (Miller & Otero, 1994), distributed from Mexico south to Argentina (Bryk, 1930). The tribe is placed in the subfamily Dioptinae (400 species), containing mostly diurnal forms, which represents a highly derived lineage within the Notodontidae (Miller, 1991, 1992a). Monophyly of the Josiini has been established based on a unique configuration of the metathoracic hearing organ in adults (Sick, 1940; Miller & Otero, 1994; see also Fig. 8 below). All josiines are diurnal, and the species are involved in mimicry complexes with taxa from numerous unrelated moth and butterfly families (Hering 1925; Seitz, 1925; K¨ohler, 1930). Furthermore, members of the tribe exhibit a wide variety of colourful, apparently aposematic wing patterns (Fig. 1), providing an opportunity to examine the evolution of divergent pattern types. Here, I present the first comparative analysis of josiine morphology and the first cladogram for the group. The study documents character variation throughout the clade, emphasizing the importance of using characters from all life stages. When wing pattern characters are mapped onto the josiine phylogeny, a case of M¨ullerian mimicry that had gone unrecognized by taxonomists for over 150 years is revealed.

METHODS

Species sample The species of Josiini included in this study were chosen on the basis of a single criterion — availability of freshly preserved specimens of immature stages. It had previously been found that the larval and pupal stages provide crucial character information for elucidating relationships (Miller, 1991, 1992a). The list of 24 josiine study species (Table 1) represents approximately 23% of the total for the tribe, and five of the 11 genera (Table 2). With the exception of those for three species, all immatures were collected by the author and his colleagues on field trips, made during the last four years, to Costa Rica, Panama, French Guiana, Venezuela and Ecuador. Immature stages have previously been described for only five of these species (Miller & Otero, 1994). Of the six genera of Josiini not treated here, most are small — containing either one or two species (Table 2) — and extremely rare. The only relatively large genus not represented is Scea Walker, with 12 species known from the Andes of western and southern South America (Bryk, 1930). Scea is thought to be closely related to Cyanotricha Prout and Thirmida Walker (Prout, 1918; Hering, 1925), two genera that are included in the study sample. Hostplants Like the adults, josiine caterpillars are brightly coloured (Fig. 2). Despite being conspicuous, very little has been reported on the larvae, and published hostplant records are rare (Miller & Otero, 1994). All were thought to be restricted to Passiflora (Passifloraceae), a plant genus containing over 500 species, approximately 450 of which are Neotropical (Heywood, 1979). However, recent field studies by the author

4

J. S. MILLER

PHYLOGENY OF THE JOSIINI

5

and his colleagues have uncovered the first josiines associated with plants outside the Passifloraceae. These findings are reported here for the first time. In French Guiana (April, 1994), caterpillars of Josia megaera (Figs 2, 19A–C) were discovered feeding on Turnera odorata LC Richard (Turneraceae) (Fig. 2), an aromatic shrub occurring in disturbed habitats throughout Central and South America. Josia megaera, including its various named forms and subspecies, occurs from Mexico south to Bolivia and east to the Guianas and the state of Bahia, Brazil (Bryk, 1930). Since TABLE 1. Species included in the analysis. Species distributions based on unpublished label data from museum specimens Ingroup Taxon

Collecting locality

Collector(s)

Cyanotricha necyria Felder Getta baetifica Druce Josia annulata Dognin Josia auriflua Walker Josia aurifusa Walker Josia cruciata Butler Josia draconis Druce Josia ena Boisduval Josia flavissima Walker Josia fluonia Druce Josia fornax Druce Josia frigida Druce Josia gigantea Druce Josia gopala Dognin Josia insincera Prout Josia interrupta Warren Josia ligata Walker Josia megaera Fabricius Josia radians Warren Josia striata Druce Josia turgida Warren Polyptychia fasciculosa Felder Thirmida discinota Warren Thirmida superba Druce

Sigchos, Ecuador 1 Tinalandia, Ecuador 2 El Amparo, Venezuela 3 Tinalandia, Ecuador 2 Choroní, Venezuela 4 Cerro Campana, Panamá 5 Canal Zone, Panamá 6 Matoury, French Guiana 7 Rancho Grande, Venezuela 3 El Angel, Ecuador 8 Rio Pastaza, Ecuador 2 Chepo, Panamá 6 Chompipe, Costa Rica 6 Mérida, Venezuela 3 Pte. Victoria, Venezuela 3 Tinalandia, Ecuador 9 El Angel, Ecuador 8 Coralie, French Guiana 7 Mérida, Venezuela 3 Las Pampas, Ecuador 2 Barinas, Venezuela 3 Coralie, French Guiana 7 Mérida, Venezuela 3 Cosanga, Ecuador 100

Colombia S to Peru Honduras S to Ecuador Venezuela, Colombia S to Peru Colombia S to Peru Venezuela Costa Rica, Panamá Panamá S to Brazil, Bolivia Panamá, Trinidad S to Brazil Venezuela, Ecuador Ecuador Ecuador Mexico S to Panamá Costa Rica, Panamá Venezuela Venezuela Colombia, Ecuador Colombia S to Peru Mexico S to Bolivia, Brazil Venezuela, W Colombia Ecuador Venezuela Colombia E to Brazil Venezuela Ecuador

Outgroup taxon: Zunacetha annulata Guérin Tithraustes haemon Druce

Mérida, Venezuela Nusagandi, Panamá

Costa Rica S to Venezuela Costa Rica, Panamá

3 6

Distribution

COLLECTOR(S): 1 – E. Tapia (Quito, Ecuador) 2 – J.S. Miller, L.D. Otero, E. Tapia 3 – L.D. Otero (Mérida, Venezuela) 4 – J.S. Miller & L.D. Otero 5 – J.S. Miller 6 – C. Snyder (AMNH) 7 – J.S. Miller, C. Snyder, L.D. Otero, B. Hermier 8 – R. Friesen (USDA, Hilo, Hawaii) 9 – J.E. Rawlins (CMNH) 10 – J.S. Miller & E. Tapia

Figure 1. Study species (Figs A–X, Josiini; Figs Y, Z, outgroup taxa; Figs A'–E', mimics). A, Josia megaera; B, J. interrupta; C, J. insincera; D, J. frigida; E, J. ligata; F, J. radians; G, J. gigantea; H,J. turgida; I, J. aurifusa; J,J. auriflua; K, Polyptychia fasciculosa; L, Getta baetifica: M, Cyanotricha necyria; N, Thirmida superba; O, T. discinota; P, Josia draconis; Q, J. ena; R, J. fornax; S, J. flavissima; T, J. gopala; U, J. fluonia; V, J. annulata; W, J. cruciata; X, J. striata; Y,Tithraustes haemon; Z; Zunacetha annulata; A', Chrysophila sp. (Pyralidae; Costa Rica); B', Desmotricha ursula Cramer (Arctiidae; French Guiana); C', Mesenopsis melanochlora Godman & Salvin (Riodinidae; Peru); D', Josiomorpha penetrata Walker (Arctiidae; Mexico).

6

J. S. MILLER

TABLE 2. Genera included in the Josiini (following Bryk, 1930; Miller & Otero, 1994), and the number of species from each genus used in this study Genus Anticoreura Prout Cyanotricha Prout Getta Walker Josia Hübner Leptactea Prout Mitradaemon Butler Phavaraea Walker Polyptychia Felder Scea Walker Scedros Walker Thirmida Walker Total

No. of spp. included

No. of spp. examined

1 2 3 68 1 5 2 2 12 1 6 103

0 1 1 19 0 0 0 1 0 0 2 24

the initial discovery, Josia megaera has been recorded on Turnera in Venezuela (L.D. Otero; July, 1994), and it is likely that this genus serves as hostplant throughout the moth’s range. These clues led to the discovery of another josiine on Turneraceae. Josia draconis (Fig.1) is a fairly common species in the Canal Zone of Panam´a (Forbes, 1939), and we had collected adults with considerable regularity on previous trips. Its hostplant, however, remained a mystery. In June, 1994, larvae of J. draconis (Fig 19D–F) were found (by C. Snyder, AMNH) on Turnera panamensis Urban at a site near Barro Colorado Island. Turnera panamensis, a shrub of 1–4 meters, is known only from lowland moist forests in the states of Panam´a and Colon (Croat, 1978). All the other josiines in the study sample were collected on Passiflora. Killip (1938) divided the American Passiflora species into 22 subgenera, and Josiini have so far been recorded from six of those (J.S. Miller, unpubl.). The majority of hostplants are in the subgenera Passiflora or Plectostemma. However, two of the moths included in this study feed on plants in the subgenus Astrophea: Caterpillars of Getta baetifica (Figs 1, 2) were discovered in Ecuador (May, 1993) on Passiflora macrophylla and P. arborea, and larvae of Polyptychia fasciculosa (Fig. 2) were collected on P. candida in French Guiana (April, 1994). Members of Astrophea are unique in that they grow as woody shrubs and trees (Fig. 2) rather than as vines (Killip, 1938; Holm-Nielsen, Jorgensen & Lawesson, 1988), one of the features making this purportedly the most primitive element within the genus Passiflora (Benson, Brown & Gilbert, 1976). The families Passifloraceae, Turneraceae and Violaceae are closely related, being placed together in the order Violales (Cronquist, 1988; Gentry, 1993). Among the few dioptine host associations known outside the Josiini, larvae from several genera have been recorded on Rinorea and Hybanthus in the Violaceae (Wolda & Foster, 1978; Miller, 1992b, unpubl. data). Hostplant relationships among Josiini and their relatives show a remarkable parallel with those of the butterfly tribe Heliconiini (Nymphalidae: Heliconiinae). As currently defined (Harvey, 1991), the Heliconiini contains approximately 150 species. The majority of these feed as larvae on Passiflora

PHYLOGENY OF THE JOSIINI

Figure 2. Final instar larvae and hostplants of the study species. Top left, outgroup species Zunacetha annulata (Venezuela) on its host, Hybanthus (Violaceae); Top right, Josia megaera on its host, Turnera odorata (Turneraceae); Second row left, Josia flavissima (Venezuela); Second row right, Josia fornax (Ecuador); Third row left, Polyptychia fasciculosa (French Guiana); Third row right, Getta baetifica (Ecuador); Bottom row left, young plant of Passiflora candida (Subgenus Astrophea), host of P. fasciculosa; Bottom row right, flower of Turnera odorata (French Guiana).

7

8

J. S. MILLER

or Violaceae (Ackery, 1988), and species in two genera — Euptoieta and Eueides — use Turneraceae (Janzen, 1983; DeVries, 1987). This convergence in host use between Josiini and Heliconiini probably reflects similar adaptations to plant secondary chemistry. Passiflora species contain cyanogenic glycosides and harmane alkaloids (Spencer, 1988), chemicals that have been isolated from the tissues of heliconiines (Nahrstedt & Davis, 1981, 1983; Cavin & Bradley, 1988). As yet no secondary chemicals are known to be shared by the Passifloraceae, Violaceae and Turneraceae, but Ehrlich & Raven (1964), noting host association patterns among Heliconiini, predicted that they will eventually be found. Morphology The phylogenetic analyses in this paper are based on characters from the external morphology of adults, larvae and pupae. Adult specimens were prepared by removing the wings and soaking the body in hot 10% KOH for approximately 10 min. Scales and tissues were then removed with a fine brush, and each preparation was placed in a vial of 70% ethanol. Male and female genitalia were removed from the alcohol-preserved specimens, stained with Chlorazol Black, and mounted on slides in Canada Balsam. Wings were stained in Eosin Y and mounted in balsam for study of venation patterns. Larval and pupal specimens were from reared material preserved in 70% alcohol. To prepare larval parts for scanning electron microscopy, they were dissected from the body, critical-point dried, and mounted on stubs using carbon tape. Morphological terms follow recent general treatments of the Lepidoptera (Common, 1990; Nielsen & Common, 1991; Scoble, 1992). Particularly useful sources for adults are Ehrlich (1958) and Oseto & Helms (1976). Terminology for larval structures follows Peterson (1962) and Stehr (1987), while that for pupae follows Mosher (1916). Background information concerning morphology of the Notodontidae is presented in Miller (1991, 1992a), while Miller & Otero (1994) provide a detailed study of larvae and pupae for the Josiini. Morphological drawings were made using a camera lucida attached to a Zeiss SV8 stereomicroscope. Electron micrographs were taken with a Zeiss DSM 950 Digital Scanning Microscope. Cladistic analyses The 24 species of Josiini were surveyed for morphological character variation along with two dioptines, Zunacetha annulata and Tithraustes haemon, designated as outgroup taxa. Both outgroup species belong in a clade, defined by presence of a stridulatory organ in the male forewing (Forbes, 1922, 1939; Miller, 1989), that is the likely sister group of the Josiini (J.S. Miller, unpubl. data). Zunacetha annulata, occurring from Mexico south to Venezuela, feeds on Hybanthus in the Violaceae (Wolda & Foster, 1978), while Tithraustes haemon, from Costa Rica and Panama, feeds on understory palms (C. Snyder & J.S. Miller, in prep.). Data were analysed using the Hennig86 (Farris, 1988) and PAUP programs (Swofford, 1991). In Hennig86, the search for maximum parsimony trees used the

PHYLOGENY OF THE JOSIINI

9

‘m*’ and ‘bb*’ options. In PAUP, a heuristic search was performed with 10 randomaddition sequences. The search terminated with the same tree all 10 times. In both analyses, multistate characters were treated as either ordered or unordered, depending on which transformation hypothesis was most plausible. Trees were rooted using the two outgroup taxa together.

RESULTS

Cladograms A total of 86 characters, 49 binary and 37 multistate, were identified (Table 3), including 59 from adults, 23 from larvae, and four from pupae. Altogether there were 215 character states. The majority of adult traits were from male and female genitalia. Analyses by both Hennig86 and PAUP produced a single mostparsimonious tree (Fig. 3) with a length of 209, a retention index of 0.84, and a consistency index of 0.63. The cladogram forms the basis for all subsequent discussion. Clade numbers referred to in the text follow those in Figure 3. The data matrix appears in Appendix 1. Several important features of this cladistic result should be mentioned. First, there is strong support for monophyly of the Josiini (Clade 1). Whereas only a single synapomorphy was known before (the modified tympanum, Character 16), the new list includes nine derived traits of immatures and adults (Table 4). Second, Josia megaera, one of the Turnera-feeders, is apparently the most primitive member of the Josiini. However, the other Turnera-feeder, Josia draconis (Clade 15), is more derived and is not closely related to J. megaera. Third, this cladogram demonstrates that the classification of Josia, as previously conceived (e.g. Bryk, 1930; Forbes, 1931), is untenable. In addition to J. megaera being alone at the bottom of the phylogeny, members of what are now called Josia appear in Clades 3, 15 and 17 (Fig. 3).

Morphology In an attempt to present the morphological data as concisely as possible, I limit the discussion to broad patterns of character evolution within the Josiini. Details of the analysis can be retrieved from examination of the figures and the list of traits. Each character state is indicated in the figures using the corresponding number from Table 3. For example, in Figure 4A, number ‘1.1’ refers to character 1, character state 1: ‘labial palpus segment 2 shorter than segment 1’ (Table 3). I have attempted to illustrate all apomorphies with the exception of those already adequately documented in recent publications. In the case of the latter, references are cited. Adults Informative head characters were found in the shape of the labial palpus, in eye size, and in the antenna. Eye size, supposedly correlated with nocturnal as opposed to diurnal behaviour in other Lepidoptera (Powell, 1973; Ferguson, 1985; Miller, 1991), varies greatly within Josiini (Fig. 4). However, all species are apparently diurnal so the reason for this variation is unknown. A previously undescribed josiine

10

J. S. MILLER

synapomorphy is an unusual configuration at the junction of each pectination with the shaft of the antenna in males (Character 4, Fig. 5). Differences in wing venation form the basis for the generic classification of the Dioptinae (Prout, 1918; Hering, 1925). The vast majority of species exhibit venation typified by that in Figure 6A (Miller, 1987). Notably, veins M3 and CuA1 are stalked in the fore and hindwings, and forewing vein M1 arises from the base of the radial sector. Within the Josiini there is variation in the exact position of forewing vein M1, and in the size of the forewing discal cell (Figs. 6, 7). A highly reduced male forewing discal cell, associated with a stridulatory organ (Miller, 1989), occurs among outgroup species (Characters 8, 9), but this structure is not found in Josiini. The only josiines with a reduced male forewing discal cell are

Figure 3. Cladogram produced by analysis of 86 morphological characters; length = 209, CI = 0.63, RI = 0.84.

PHYLOGENY OF THE JOSIINI

11

TABLE 3. Morphological characters used in the analysis. Those treated as ordered are designated by a ‘[+]’ following the character description; those treated as unordered have a ‘[–]’ ADULTS: 1. Segment 2 of labial palpus as long as, or longer than, segment 1 [0]; segment 2 shorter than segment 1 (Fig. 4A) [1]; segment 2 curved upward near middle (Fig. 4B) [2]. [–] 2. Segment 3 of labial palpus moderately long relative to segment 2 [0]; LP segment 3 elongate (Fig. 4E) [1]; LP segment 3 short, bullet-shaped (Fig. 4D) [2]. [–] 3. Eyes relatively large, postgena (in lateral view) fairly broad (Fig. 4D) [0]; eyes extremely large and round, extending below hypostoma, postgena a narrow strip (Fig. 4C) [1]; eyes small, postgena broad, hypostomal area exposed (Fig. 4A) [2]. [–] 4. Pectinations broadly joined to each antennal segment (Figs. 5A, 5B) [0]; antennal pectinations thinly joined to each segment, each pectination appearing hinged (Fig. 5C–F) [1]. 5. Male antennae pectinate almost to apex, less than 12 terminal segments simple [0]; male antennae wth more than 15 distal segments simple (Fig. 1) [1]. 6. Male antennal pectinations relatively long, antennal shaft evenly tapered (Fig. 5C) [0]; antennal pectinations short, appressed to antennal shaft, shaft wider in middle portion (Fig. 5F) [1]. 7. Forewing vein M1 arising from discal cell (Fig. 6C) [0]; FW M1 arising from upper angle of DC near base of Rs (Fig. 6B) [1]; M1 arising from Rs near base of R2 (Fig. 7E) [2]; M1 arising on Rs beyond base of R2 (Fig. 7D) [3]. [+] 8. Male forewing discal cell not reduced (Fig. 6D) [0]; male FW DC reduced, less than 1/2 wing length (outgroup; Miller, 1989) (Fig. 6C) [1]. 9. Veins M1 and M2 in male forewing not swollen [0]; veins M1 and M2 in male FW swollen beyond discal cell (outgroup; Miller, 1989) [1]. 10. Hindwing venation not reduced (Fig. 6D) [0]; HW venation reduced, veins Rs and M1 completely anastomosed, DC small (Fig. 6C) [1]. 11. Forewing costa dark [0]; FW costa orange in proximal half (Figs. 1, 7D) [1]. 12. Forewing pattern various, without a transverse band [0]; FW with a transverse band beyond DC (Figs 6A, 6C, 7A) [1]; FW with a transverse band encompassing end of DC (Fig. 6D) [2]. [+] 13. Forewing without a longitudinal stripe [0]; FW with a longitudinal stripe from base, not extending beyond end of DC (Fig. 6A) [1]; longitudinal FW stripe extending beyond DC almost to wing margin (Figs 6B, 7C, 7D) [2]. [+] 14. Hindwing margin completely orange or completely black [0]; HW margin orange with a black dash from outer edge (Fig. 1) [1]. 15. Lateral portion of hind tibia dark [0]; lateral portion of hind tibia buff [1]; lateral portion of hind tibia with a thin line of buff-colored scales [2]. [+] 16. Metathoracic tympanal cavity not completely enclosing tympanal membrane (Miller, 1987, 1989) [0]; tympanal cavity forming a deep, kidney-shaped pocket that encloses tympanal membrane (Fig. 8A)[1]; pocket large and kettle drum-shaped, with a small lateral opening (Fig. 8B) [2]. [+] 17. Metepimeron almost completely covered with scales [0]; metepimeron scaleless and heavily sclerotized in region posterior to opening of tympanal cavity [1]. 18. Base of abdomen dark above (Fig. 1) [0]; base of abdomen with a broad orange transverse band dorsally (Fig. 1) [1]. 19. Abdomen white below, no distinct lateral stripes [0]; abdomen white below, with orange or yellow lateral stripes (Forbes, 1931) [1]; abdomen uniformly dark [2]; abdomen uniformly dark except for slightly lighter lateral stripes (Miller & Otero, 1994) [3]. [+] 20. Lateral stripes on abdomen either absent or concolorous for entire length [0]; lateral stripes yellow, changing to buff on segment 8 (Miller & Otero, 1994) [1]. MALE TERMINALIA: 21. Anterior apodeme on sternum 8 broad, rounded (Fig. 9F) [0]; AA on S8 acute at apex (Fig. 9H) [1]; AA very long, narrow (Fig. 9G) [2]; AA broad and slightly bifid, two rounded humps (Miller, 1988) [3]; AA blunt at apex (Fig. 9C) [4]. [–] 22. Posterior margin of sternum 8 with a relatively shallow notch (Fig. 9J) [0]; posterior notch deep, lateral processes narrow (Fig. 9H) [1]; posterior notch with a medial process (Fig. 9C) [2]. [–]

12

J. S. MILLER

TABLE 3. – continued 23. Lateral processes of posterior notch on sternum 8 smooth [0]; lateral processes of S8 posterior notch spiculate (Fig. 9B) [1]. 24. Uncus not robust or compressed, without a dorsal flange [0]; uncus robust, dorso-laterally compressed (Fig. 13E) [1]; uncus with a large, dorso-medial flange (Fig. 12E) [2]. [–] 25. Socii broad [0]; socii narrow, finger-like (Fig. 11A) [1]; socii attenuated, sclerotized (Fig. 13D) [2]. [+] 26. Socii without a dorsal process [0]; each socius with a dorsal process (Fig. 12D) [1]. 27. Sides of tegumen parallel, tegumen robust [0]; tegumen wide, expanded in dorsal portion (Fig. 12D) [1]; tegumen narrow (Fig. 13A) [2]. [+] 28. Transtilla relatively small, not deeply incurved [0]; transtilla large, deeply concave (Fig. 13E) [1]. 29. Transtillar arms essentially horizonal (Figs 10A, 11D) [0]; transtillar arms downcurved (Figs 11E, 12E) [1]; transtillar arms greatly downcurved, sinnuate (Fig. 12D) [2]. [+] 30. Manica completely membranous above transtilla (Fig. 12A) [0]; a patch of short spicules in manica above transtilla (Fig. 12E) [1]; spicules long, patch heart-shaped (Fig. 13E) [2]. [+] 31. Valve not narrow (Fig. 11E) [0]; valve narrow (Fig. 11A) [1]; valve narrow, costa curved and sclerotized near apex (Fig. 11D) [2]. [+] 32. Costa of valve with medial portion simple [0]; costa with a spine-like medial process (Fig. 12A) [1]. 33. Base of costa simple [0]; base of costa with a blunt process (Fig. 12E) [1]; base of costa serrate (Fig. 13E) [2]. [+] 34. Costa of valve roughly straight, parallel-sided [0]; costa expanded near apex (Fig. 11E) [1]; costa reflexed upward (Fig. 13E) [2]. [–] 35. Costa of valve relatively long [0]; distance between base of transtilla and apex of costa short (Fig. 12E) [1]. 36. Apex of costa with a blund projection [0]; apex of costa with a sharp, thorn-like process (Fig. 11a) [1]; apex of costa with a robust, truncate process (Fig. 13D) [2]. [–] 37. Barth’s Organ relatively large, occupying approximately 1/2 of the valve (Fig. 10B) [0]; BO small, pleats sclerotized at upper angles (Fig. 12A) [1]; BO extremely large, rounded, occupying over 3/4 of the value (Fig. 13A) [2]. [+] 38. Lateral strut of vinculum narrow, connected at base of costa near junction of transtilla [0]; lateral strut of vinculum wide, connected near midpoint of costa beyond junction of transtilla (Fig. 12D) [1]. 39. Sacculus with a small internal apodeme at base or apodeme absent [0]; sacculus with a long, sclerotized internal apodeme at base (Fig. 12E) [1]. 40. Inner surface of valve without a fold at base of sacculus [0] a well-defined fold on inner surface of valve at base of sacculus, enclosing hairlike scales (Fig. 10B) [1]; a loose fold at base of sacculus (Fig. 12A) [2]. [+] 41. Hair-like androconia present on inner surface of valvae [0]; short, tubular androconia on ventral inner surface of valve in addition to hair-like ones (Fig. 13A) [1]. 42. Androconia on lateral surface of valve without raised bases [0]; each androconium on lateral surface of valve with a dark, raised base (Fig. 12D) [1]. 43. Ventromedial junction of valvae smoothly convex, sometimes concave (Fig. 12A) [0]; ventromedial junction of valve quadrate with elbow-like lateral angles (Fig. 13E) [1]; ventromedial junction of valve sinnuate, projecting slightly at midline (Fig. 12E) [2]; [+] 44. Aedeagus essentially cylindrical (outgroup) [0]; aedeagus rounded, bulbous (Fig. 12B) [1]; aedeagus slightly compressed dorsoventrally, apex sclerotized (Fig. 11C) [2]. [+] 45. Base of aedeagus smoothly rounded [0]; base of aedeagus with a pair of wing-like processes near opening of simplex (Fig. 13C) [1]; base of aedeagus horn-shaped (Fig. 12C) [2]. [–] 46. Apex of aedeagus narrowing to a straight point (Fig. 13B) [0]; point at apex of aedeagus curved laterally (Fig. 11B) [1]. 47. Vesica moderately long [0]; vesica at least twice as long as aedeagus (Fig. 11F) [1]; vesica relatively short and bulbous (Fig. 12B) [2]; vesica relatively long, curving anterodorsally (Fig. 13F) [3]; vesica large, opening on dorsum near base (Fig. 12C) [4]. [–]

PHYLOGENY OF THE JOSIINI

13

TABLE 3. – continued 48. Vesica with distal cornuti approximately equal in size to others [0]; vesica with one distal cornutus greatly enlarged (Fig. 11C) [1]; distal cornutus club-shaped (Fig. 13C) [2]; two distal cornuti greatly enlarged (Fig. 10A) [3]. [–] 49. Cornuti scattered on vesica or restricted to distal portion [0]; cornuti concentrated in a line on posterior surface of vesica (Fig. 13F) [1]. FEMALE GENITALIA: 50. Female T8 sclerotized, joined across dorsum (Fig. 14D) [0]; female T8 reduced to two thin straps, completely membranous dorsomedially (Fig. 15C) [1]. 51. Margins of papillae anales rounded (Fig. 15D) [0]; papillae anales slightly emarginate (Fig. 15C) [1]; PA’s strongly emarginate, with a distinct dorsal lobe (Fig. 14C) [2]. [+] 52. Postvaginal plate not large or modified [0]; PVP broad, sclerotized, wrinkled, with a membranous medial portion and posterior excavation (Fig. 14B) [1]. 53. Ductus bursae not elongate [0]; ductus bursae long and narrow, mostly membranous (Fig. 15A) [1]; ductus bursae vase-shaped, sclerotized (Fig. 15C) [2]. [–] 54. Ductus bursae without internal spines [0]; ductus bursae with internal spines (Fig. 15B) [1]; ductus bursae with a small patch of short spicules only (Fig. 14B) [2]. [+] 55. Corpus bursae with a relatively small sclerotized region near base (Fig. 14E) [0]; entire basal half of corpus bursae broadly sclerotized (Fig. 15C) [1]; entire corpus bursae membranous (Fig. 14B) [2]. [–] 56. Base of corpus bursae with a spiculate, horn-shaped process (Fig. 14E) [1]; no horn-shaped process [0]. 57. Corpus bursae with spines or membranous at base, not wrinkled [0]; corpus bursae with a wrinkled, sclerotized region near base (Fig. 15A) [1]. 58. Junction of ductus seminalis and corpus bursae expanded slightly, membranous (Fig. 14A) [0]; DS attached to corpus bursae by a curled projection with a sclerotized dorsal band (Fig. 15D) [1]. 59. Signum oblong, spiculate, with a medial seam (Fig. 14A) [0]; signum small, round, with long internal spines (Fig. 15A) [1]; signum ‘bird-shaped’, with long lateral internal spines, a spiculate, trianglular central portion, and an external ‘horn’ (Fig. 15C) [2]. [–] LARVAE: 60. Head orange-brown, without pattern except dark pigment surrounding stemmata (Figs 2, 19A) [0]; head white with various dark stripes or patches (Figs 2, 18A, 18B, 19D, 19E) [1]; head entirely black except ecdysial suture, ecdysial lines and clypeus (Miller & Otero, 1994) [2]. [+] 61. Head surface finely spiculate (Fig. 16A, B) [0]; head surface smooth (Fig. 16C) [1]. 62. Frons rounded at apex, a compressed triangle (Fig. 19B) [0]; frons acute at apex, taller (Fig. 19E) [1]. 63. Spinneret narrow, tapered distally (Fig. 16E) [0]; spinneret wide, truncate distally (Fig. 16D, F) [1]. 64. Antennal segment 2 approximately twice the length of segment 1 (Fig. 17A) [0]; antennal segment 2 less than twice the length of segment 1 (Fig. 17B) [1]. 65. Antennal segment 2 parallel-sided (Fig. 17A) [0]; antennal segment 2 expanded distally (Miller & Otero, 1994) [1]. 66. Surface of antenna smooth (Fig. 17B) [0]; surface of antenna rugose (Fig. 17C) [1]. 67. Segment 3 of maxillary palpus not elongate (Fig. 17D) [0]; segment 3 of maxillary palpus elongate (Fig. 17E) [1]. 68. Prothoracic shield completely pigmented medially (Fig. 18F) [0]; PS with a wide, unpigmented medial seam (Fig. 18D) [1]; PS with a narrow, or partial, medial seam (Fig. 18E) [2]. [–] 69. Prothoracic shield pigmented, brown to brownish black (Fig. 18D–F) [0]; PS unpigmented, dorsum of prothorax completely white (Fig. 2) [1]. 70. Setal bases on prothoracic shield not darkly pigmented (Fig. 18E) [0]; setal bases on PS darkly pigmented (Fig. 18F) [1]. 71. Anterolateral angles of prothoracic shield rounded, not produced forward (Fig. 18D) [0]; anterolateral angles of PS produced slightly forward (Fig. 18F) [1]; anterolateral angles of PS curled inward (Fig. 18E) [2]. [+]

14

J. S. MILLER

TABLE 3. – continued 72. Tarsal seta 3 leaf-shaped, apex acute (Fig. 17F) [0]; TS3 leaf-shaped, apex emarginate (Miller & Otero, 1994) [1]. 73. Segments T2, A1, A3, A5 and A8 more darkly pigmented than alternating ones (Fig. 2) [0]; all segments equally pigmented (Fig. 2) [1]. 74. Pattern various [0]; light-coloured lateral areas on body roughly rectangular, simple (Miller & Otero, 1994) [1]; light-coloured areas reticulate (Miller & Otero, 1994) [2]; lateral pattern a complex series of broken, longitudinal stripes (Fig. 18A) [3]; light-coloured areas expanded, dark dorso-lateral patches reduced to irregular cross-stripes (Fig. 19D) [4]. [–] 75. Venter concolorous [0]; venter with reddish-maroon transverse lines on A1 and A2 (Miller & Otero, 1994) [1]. 76. Segment A9 white and reddish maroon, not conspicuous (Miller & Otero, 1994) [0]; A9 conspicuous, almost completely white dorsally (Fig. 19F) [1]. 77. Only three L setae on segments A2–A6 (Fig. 18A) [0]; A2–A6 with a fourth L seta posterior to spiracle (Miller & Otero, 1994) [1]. 78. Seta SV2 on A2 located below a line drawn between L3 and SV3 (Fig. 18A) [0]; SV2 on A2 on a line horizontal with L3 and SV3 (Miller & Otero, 1994) [1]. 79. Anal plate with 4 setae on each side (Figs 19F, 20B) [0]; AP with more than 4 setae on each side (Figs 18C, 20C) [1]. 80. Lateral plate of A10 proleg with 4 or 5 setae (Fig. 20D) [0]; LP of A10 proleg with 9 to 14 setae (Fig. 20E) [1]; LP of A10 proleg with 15 or more setae (Fig. 20F) [2]. [+] 81. Primary setae not thickened [0]; primary setae thick and fleshy (Miller & Otero, 1994) [1]. 82. Segment A9 with six setae on each side (Fig. 19C) [0]; A9 with an extra primary seta in the L2 region (Fig. 19F) [1]. PUPAE: 83. Cremaster with 8 to 10 hook-shaped setae (Fig. 21D) [0]; cremaster with more than 12 hook-shaped setae (Fig. 21E, F) [1]. 84. Cremaster setae robust (Fig. 21A) [0]; cremaster setae delicate (Fig. 21D) [1]. 85. Striae at apex of cremaster (dorsal view) either irregular or in longitudinal lines (Fig. 21E) [0]; striae at apex of cremaster forming two or three concentric rings (Fig. 21A) [1]. 86. Dorsum of abdominal segments smooth, lacking setae (Fig. 21A) [0]; dorsum of abdomen with two rows of setae (Fig. 21E; Miller, 1987, 1992a) [1].

TABLE 4. Synapomorphies for the Josiini 1. Male antennae with pectinations thinly joined to each segment, pectinations appearing hinged (Fig. 5C–E). 2. Metathoracic tympanal cavity forming a deep pocket that encloses tympanal membrane (Fig. 8). 3. Socii of male genitalia narrow, finger-like (Figs 10–13). 4. Aedeagus rounded at base, with a simple distal point (Figs 10–13). 5. Female tergum 8 reduced to two thin straps, completely membranous dorsomedially (Fig. 15C). 6. Larval head surface smooth (Fig. 16C). 7. Larval spinneret wide, truncate distally (Fig. 16F). 8. Four larval instars rather than five (Miller & Otero, 1994). 9. Pupa with dorsum of abdominal segments smooth, lacking specialized setae (Fig. 21A).

PHYLOGENY OF THE JOSIINI

15

Getta and Polyptychia (Clade 16). These have a reduced hindwing cell as well. Here, reduction appears to be correlated with the presence of specialized androconial patches, which occur on the upper surface of the hindwing in P. fasciculosa (Fig. 6C), and on the underside of both the fore- and hindwings in G. baetifica. The taxa bearing androconia on the wings, Getta and Polyptchia, are the same ones whose larvae feed on the primitive Passiflora subgenus Astrophea (see ‘Hostplants’ above). Interestingly, males of P. fasciculosa are unique among Dioptinae in having androconia on the hind legs as well.

Figure 4. Heads of representative Josiini (males) in lateral view, anterior at left (Characters 1–3). Only the base of the haustellum is shown. A, Cyanotricha necyria; B, Getta baetifica; C, Josia insincera; D, Josia fornax; E, Josia megaera. An = antennal socket; E = eye; H = haustellum; Hs = hypostoma; Lp = labial palpus segment; Pt = postgena. Scale bars = 0.5 mm.

16

J. S. MILLER

Figure 5. Scanning electron micrographs of male antennae, ventral surface (Characters 4, 6). A, Tithraustes haemon (90X); B, T. haemon (368X); C, Cyanotricha necyria (98X); D, C. necyria (394X); E, C. necyria (761X); F, Josia flavissima (134X)

PHYLOGENY OF THE JOSIINI

17

In butterflies, the evolution of diurnal behaviour has led to a dramatic change in intraspecific signalling systems. Nocturnal moths rely on female-dispersed chemicals for mate location, while butterflies use visual signals (Scoble, 1992). It has been argued that the high level of wing pattern convergence in butterflies, resulting from selection for mimicry, has lowered the effectiveness of visual signals (Vane-Wright & Boppr´e, 1993). As a consequence, many butterflies use close-range male chemicals,

Figure 6. Wings of Josiini (Characters 7–14); FW length in parentheses. A, Josia megaera male (14 mm); B, J. insincera female (15 mm); C, Polyptychia fasciculosa male (21mm); D, Josia ena male (12 mm). A = anal vein; CuA = cubital vein; DC = discal cell; M = medial vein; R = radial vein; Sc = subcostal vein. Melanic areas are indicated by shading.

18

J. S. MILLER

dispersed by androconia, during courtship. Getta and Polyptychia species have apparently undergone analogous modifications. Josiine wing coloration and wing pattern vary greatly (Fig. 1). The forewing can have a longitudinal stripe (Figs 6B, 7C, 7D) or a transverse band (Figs 6C, 6D, 7A). In Josia megaera (Fig. 6A) both patterns are present, while the forewing stripe of J. fluonia (Fig. 7B) seems to be intermediate between a longitudinal and a transverse one. Much of this variation was pointed out by Forbes (1931), who used the colour of the forewing costa (Character 11) and the configuration of the forewing markings in his

Figure 7. Wings of Josiini (Characters 7–14); FW length in parentheses. A, Josia draconis male (17 mm); B, J. fluonia male (14 mm); C, J. cruciata male (16 mm); D, J. frigida female (17 mm); E, J.ligata female forewing radial and medial veins (19 mm). M = medial vein.

PHYLOGENY OF THE JOSIINI

19

key to Josia species. Detailed treatment of wing pattern traits is reserved for the Discussion section (below). Colour patterns on the legs, thorax and abdomen provide important characters for separating species and for defining species groups. Forbes (1931), noting differences between species in presence or absence of abdominal stripes (Characters 18–20), regarded this as a character of considerable importance within the group. A unique kettle-drum shaped metathoracic tympanum (Richards, 1932; Sick, 1940; Kiriakoff, 1950; Miller & Otero, 1994) occurs throughout the Josiini (Character 16). This consists of a large, rounded internal pocket of the epimeron, with a horizontally-oriented tympanal membrane (Fig. 8B). Josia megaera is unique in having a smaller, kidney-shaped pocket (Fig. 8A). I have scored the latter as intermediate between the shallow pocket of the outgroup (figured in Miller, 1987, 1989), and the large one of other Josiini. A character seemingly associated with size of the tympanal pocket is the presence or absence of scales on the metepimeron posterior to the tympanal opening. In species with a large kettle-drum these scales are absent, while in J. megaera and the outgroup scales are present. The function of such a highly developed tympanum in josiines is unclear. The usual explanation for its purpose in other Noctuoidea is detection of bat predators (Fullard, 1984), but this is certainly not the case for diurnal taxa such as the Josiini. A more likely possibility is that the tympanum is used for intraspecific communication, perhaps during mating. Male genitalia, recognized as crucial for clarifying Lepidoptera species relationships, have never been studied in Josiini. They provide a wealth of characters. In Notodontidae, male segment 8, along with the valvae, aids in clasping the female (Miller, 1988). Accordingly, sternum 8 is greatly elaborated and can be used to separate closely related species (e.g. Franclemont, 1946; Todd, 1973; Weller, 1990, 1991). Male Josiini exhibit modifications on both the anterior and posterior sternal margins (Fig. 9). Perhaps the most conspicuous feature of the male genitalia is the heavily pleated sacculus (Fig. 10–13) bearing a prominent brush of long, deciduous

Figure 8. Metathoracic segment of adult Josiini (males) in lateral view, anterior at left (Character 16). A. Josia megaera; B. J. ena. A = subalare; B = metascutal bulla; C = coxa; Em = epimeron; Es = episternum; M = metameron; S = metascutellum. Scale bars = 0.5 mm.

20

J. S. MILLER

Figure 9. Eighth sternum of male Josiini in ventral view, posterior margin at top (Characters 21–23). A, Josia megaera; B, J. insincera; C, J. radians; D, J. aurifusa; E, J. ena; F, Thirmida discinota; G, Getta baetifica; H, Josia fornax; I, J. gopala; J, J. annulata. Scale bars = 0.5 mm.

PHYLOGENY OF THE JOSIINI

21

androconia (not shown in all figures). This structure, termed Barth’s Organ (Weller, 1989) after the researcher who described it (Barth, 1955), occurs throughout the Notodontidae (Miller, 1991) and is well developed in the outgroup species used here (male genitalia of Zunacetha annulata shown in Miller, 1991: fig. 329). Although the organ is presumed to function during courtship, evidence is lacking. Within the Josiini the sacculus shows varying degrees of development, and a multiplicity of scale types is associated with it; different species show lateral, mesal and ventral androconia with different scale sizes and shapes (e.g. Figs 12D, 13A). Androconial structure was not analysed in detail. Some josiines with a large Barth’s Organ also have an internal apodeme (Character 39, Fig. 12E) that presumably attaches to a muscle which could serve to fold and unfold the sacculus. A great deal of character information was found in the shape of the valve and aedeagus, especially the vesica

Figure 10. Genitalia of Josia megaera. A, Aedeagus of male in lateral view, anterior at left; B, Male genitalia in posterior view, aedeagus removed; C, Female genitalia in lateral view, anterior at left. Ae = aedeagus; BO = Barth’s Organ; C = costa; Cb = corpus bursae; Cn = cornutus; Db = ductus bursa; O = ostium; Pa = papillae anales; Pv = postvaginal plate; Sc = socius; Sg = signum; Tg = tegumen; Ts = transtilla; U = uncus; V = valve; Vn = vinculum; Vs = vesica; Scale bars = 1.0 mm.

22

J. S. MILLER

Figure 11. Male Josiini, with genitalia in posterior view and aedeagus in lateral view (Characters 24–49). A, Josia radians genitalia; B, J. radians aedeagus; C, J. aurifusa aedeagus; D, J. aurifusa genitalia; E, Thirmida discinota genitalia; F, T. discinota aedeagus. Scale bars = 1.0 mm.

PHYLOGENY OF THE JOSIINI

Figure 12. Male Josiini, with genitalia in posterior view and aedeagus in lateral view (Characters 24–49). A, Josia insincera genitalia; B, J. insincera aedeagus; C, Getta baetifica aedeagus; D, G. baetifica genitalia; E, Josia ena genitalia; F, J. ena aedeagus. Scale bars = 1.0 mm.

23

24

J. S. MILLER

Figure 13. Male Josiini, with genitalia in posterior view and aedeagus in lateral view (Characters 24–49). A, Josia fornax genitalia; B, J. fornax aedeagus; C, J. gopala aedeagus; D, J. gopala genitalia; E, J. annulata genitalia; F, J. annulata aedeagus. Scale bars = 1.0 mm.

PHYLOGENY OF THE JOSIINI

25

and cornuti of the latter. Potentially, these provide the most useful traits for separating species and for defining groups of species. The female genitalia of Josiini are also extremely variable (Figs 10C, 14, 15). A unique feature is the reduction of tergum 8. In all Josiini examined except Getta (Fig. 14D) and Polyptychia, the tergum is completely membranous. In outgroup taxa it is sclerotized (female genitalia of Zunacetha annulata shown in Miller, 1991: fig. 262). Based on the cladistic result (Fig. 3), the sclerotized T8 of Polyptychia and Getta represents a reversal to the plesiomorphic condition. Females show variation in the shape of the ovipositor lobes and in the length and degree of sclerotization of the ductus bursae. Ductus bursae length appears to be correlated with the length of the

Figure 14. Female genitalia of Josiini (Characters 50–59). A, Josia radians, ventral view; B, J. aurifusa, ventral view; C, J. ena, lateral view; D, Getta baetifica, lateral view; E, Josia insincera, ventral view. Ds = ductus seminalis. Scale bars = 1.0 mm.

26

J. S. MILLER

male vesica (compare Figs 11F and 15A). The ductus in species of Josiini almost always bears internal spines, but this trait occurs throughout the Dioptinae (J.S. Miller, unpubl.). The corpus bursae is frequently sclerotized in the basal half (Fig. 15B–D), and in members of Clade 12 there is a region of complex, sclerotized folds (Fig. 15A; Miller, 1988). Two features of the female genitalia provide evidence in support of a basic dichotomy within what has previously been recognized as the genus Josia. In J. megaera and in the nine Josia species of Clade 3, the spiculate signum is roughly

Figure 15. Female genitalia of Josiini in lateral view (Characters 50–59). A, Thirmida discinota; B, Josia annulata; C, J. fornax; D, J. gopala. Scale bars = 1.0 mm.

PHYLOGENY OF THE JOSIINI

27

oblong or round with a medial seam (Character 59; Figs 10C, 14A, B, E). In these same taxa there is a small membranous expansion at the junction of the ductus seminalis (Fig. 14D) and the ductus bursae (Character 58; Fig 14A,B). Both character states are typical of the outgroup. Species in Clade 17, representing a large portion of Josia, show very different morphology for these two characters. Here, the signum is highly modified; I term it ‘bird-shaped’ because of the paired internal clusters of spines, resembling wings (Figs 14C, 15B–D). The signum is further elaborated in having a horn-like external portion. The members of Clade 17 also differ from other Josia species in that the ductus seminalis is attached to the corpus by means of a large curled structure (Figs. 15B–D), unique among Notodontidae. These apomorphies are not the only ones that support a division within Josia, but they are perhaps the most conspicuous. Larvae Larvae of the Josiini are superficially similar in appearance (Fig. 2). It is interesting that careful comparison of body colour patterns reveals highly conservative pattern elements, analogous to those in adult wings (see above). I am unaware of previous research on colour pattern homology in lepidopteran larvae. Larval colour pattern variation was employed in a relatively simple way in the present study (Characters 73–76), but this character complex potentially provides useful traits for separating species and for defining genera and species groups. In outgroup taxa and in the majority of Josiini, segments T2, A1, A3, A5 and A8 are more darkly pigmented than the alternating ones (Fig. 2). A derived trait within Josiini is the condition where all segments are equally pigmented (Fig. 2; see also figs 67 and 68 in Miller & Otero, 1994). Characters of the larvae provide two distinctive synapomorphies for the Josiini (Table 4). (1) In all species the head surface is smooth (Fig. 16C); in live caterpillars the head has a glassy appearance. The head surface of other Dioptinae is finely spiculate (Fig 16A, B). While variation in head microsculpture occurs throughout the Notodontidae, the vast majority of taxa have a spiculate or pebbled surface (Miller, 1991). (2) The spinneret of Josiini is broad (Fig. 16F) whereas that of outgroup species is more elongate (Fig. 16E). Spinneret shape varies greatly among Notodontidae (Miller, 1991), but that of the Josiini seems to be quite uniform. A third tribal synapomorphy, first described by Miller & Otero (1994) but confirmed by a more comprehensive survey in the present work, is the occurrence of only four larval instars rather than the five or six typical of other Lepidoptera. Additional useful head characters include the relative lengths of the antennal segments, the microsculpture on the surface of the antenna (Fig. 17A–C), and the length of maxillary palpus segment 3 (Fig. 17D, E). Along with adult tympanal structure (see above), two features of the caterpillar head support placement of Josia megaera at the base of the josiine phylogeny. In outgroup species, the head is uniformly orange-brown except for a dark area surrounding the stemmata (Fig. 2; see also fig. 97 in Miller, 1991). Josia megaera is unique among Josiini in exhibiting this plesiomorphic condition (Figs 2, 19A, B). In caterpillars of other josiines, the head is either conspicuously patterned with white and dark stripes (Figs 2, 18A, 18B, 19D, 19E) or is entirely black (Miller & Otero, 1994). Shape of the larval frons gives further evidence for the unique phylogenetic position of J. megaera. The frons can be rounded at the dorsal apex, as in J. megaera (Fig. 19B) and the outgroup, or acute as in all other Josiini (Figs 18B, 19E).

28

J. S. MILLER

Figure 16. Final instar larval heads. A, Zunachetha annulata (outgroup) in frontal view, area of right stemmata (119X); B, Z. annulata, close-up of area surrounding stemmata (500 3 ) showing spiculate surface (Character 61.0); C, Josia cruciata (122 ×) in frontal view, area of right stemmata showing smooth surface (Character 61.1); D, Mouthparts of Josia megaera in frontal view (240 3 ); E, Labial complex of Zunacetha annulata (282 3 ) showing relatively narrow spinneret (Character 63.0); F, Labial complex of Josia megaera (500 3 ) showing wide spinneret (Character 63.1). Sp = spinneret.

PHYLOGENY OF THE JOSIINI

Figure 17. Final instar larvae of Josiini. A, Left antenna of Josia draconis, lateral view (235 3 ); B, Left antenna of Cyanotricha necyria, lateral view (280 3 ) showing short segment 2 (Character 64.1); C, Right antenna of Getta baetifica, frontal view (217 3 ) showing rugose surface (Character 66.1); D, Left maxillary palpus of Josia draconis, frontal view (474 3 ). E, Right maxillary palpus of J. cruciata, frontal view (400 3 ) showing elongate segment 3 (Character 67.1); F, Right protarsus of J. megaera, mesal view (270 3 ) showing tarsal setae (Character 72.0). An = antennal segment; Cl = tarsal claw; Mp = maxillary palpus segment; 2, 3, 4 = tarsal setae 2, 3 and 4.

29

30

J. S. MILLER

On the larval thorax, informative traits were found in the shape and pigmentation pattern of the prothoracic shield (Characters 68–71; Fig. 18D–F). In caterpillars of many Lepidoptera the tarsae on all three thoracic legs bear remarkable, flattened setae. The function of these is unknown. Tarsal setae show interesting shape variation among lineages of Notodontidae (Miller, 1991). In Dioptinae each seta has a shape distinctive for the subfamily (Fig. 17F), but these are essentially uniform throughout the group. Only a single, subtle difference has been discovered among josiines; the apex of tarsal seta 3 is emarginate in Josia radians (Miller & Otero, 1994) and J. ligata.

Figure 18. Final instar larvae of Josiini (Characters 68–82). A, Head, thorax and A1–A3 of Thirmida superba in lateral view; B, Head of T. superba in frontal view; C, Segments A6–A10 of T. superba in lateral view. D, Prothoracic shield of T. superba, dorsal view; E, Prothoracic shield of Josia insincera, dorsal view; F, Prothoracic shield of J. striata, dorsal view. Scale bars: A, C = 2.0 mm; B, D–F = 1.0 mm. Ab = abdominal segment; An = antenna; D = dorsal seta; Fr = frons; L = lateral seta; Mp = maxillary palpus; P = proleg; Pg = prothoracic gland; Ps = prothoracic shield; SD = subdorsal seta; Sp = spiracle; St = stemmata; SV = subventral seta; T = thoracic segment; V = ventral seta.

PHYLOGENY OF THE JOSIINI

Figure 19. Final instar larvae of Josiini (Characters 60, 62, 74–82). A, Head, thorax and A1–A3 of Josia megaera in lateral view; B, Head of J. megaera in frontal view; C, Segments A6–A10 of J. megaera in lateral view; D, Head, thorax and A1–A3 of J. draconis in lateral view; E, Head of J. draconis in frontal view; F, Segments A6–A10 of J. draconis in lateral view. Ap = anal plate. Scale bars: A, C, D, F = 2.0 mm; B, E = 1.0 mm.

31

32

J. S. MILLER

The study of larval chaetotaxy, pioneered by Fracker (1915) and Hinton (1946), has played a large part in lepidopteran higher-level classification. Primary setae, those that occur in first instars, are conservative in number and position, and a widely accepted nomenclature has been developed (e.g. Kitching, 1984). Even slight variation can provide significant phylogenetic information (Stehr, 1987; Scoble, 1992). I compared chaetotaxy among first instars of all the Josiini represented in this study and found only one variable trait. In the plesiomorphic condition, exhibited by the outgroup and the majority of Josiini, seta L2 on segment A9 is single (figured in Miller & Otero, 1994). Five species show an apomorphic state, having two setae in the L1 position. This trait (Character 82), which is also expressed in subsequent instars, occurs in the members of Clade 12 (e.g. Thirmida superba, Fig. 18C) as well as in Josia flavissima and J. draconis (Fig. 19F). Such a character distribution implies considerable homoplasy (see Fig. 3). Subprimary setae, those unique to the second through ultimate instars, are also highly conservative in position (Stehr, 1987; Nielsen & Common, 1991). The pattern of subprimary setae on the head, thorax and abdomen is relatively uniform throughout the Josiini. However, some clades are characterized by the occurrence of novel setae in particular locations. For example, both Josia gopala (fig. 67 in Miller & Otero, 1994) and J. striata have an extra L seta on abdominal segments 2–6 (Character 77). A more widespread case of variation in subprimary setal number involves the prolegs on abdominal segment 10 (Character 80). In the outgroup and in all first instars, there are only 4 or 5 setae on the lateral surface of the A10 proleg (Fig. 20D). Most later instar Josiini have between 9 and 14 setae (Fig. 20E). In a few josiine species there are more than 15 setae (Fig. 20F). The primitive state (presence of 4 of 5 setae) appears to have been secondarily derived within the tribe, occurring in all species of Clade 17 (Fig. 3). When setae are abundantly distributed over the body they are termed secondary. Secondary setae occur commonly among other subfamilies of the Notodontidae (Forbes, 1948; Miller, 1991), but Cyanotricha necyria is so far the only dioptine where this trait is known (Fig. 20A). Pupae Miller (1992a) showed that pupal characters are remarkably effective at retrieving phylogenetic relationships among subfamilies of the Notodontidae. Even though relatively few characters were found in that study (n = 24), these produced a phylogeny almost identical to one based on 174 characters of larvae and adults (Miller, 1991). Within Josiini, however, the number of useful pupal characters is few (Characters 83–85). Exceptions are cremaster shape, and the size and number of cremaster setae. Cremaster shape varies greatly, even among closely related species (Miller & Otero, 1994). Two distinct sculpturing types occur: one in which the apical striae of the cremaster are irregularly arranged (Fig. 21C), and one where these are arranged in roughly concentric rings (Fig. 21A). The hook-shaped cremaster setae, typical of Lepidoptera, usually number 8 or 10 (Fig. 21A–D). In Thirmida and Cyanotricha, however, they are numerous (Fig. 21E, F). The cremaster setae of some species in Clade 19, such as Josia striata and J. flavissima, are unusual in being relatively thin (Fig. 21B–D). Although few dioptine pupae have been studied, most exhibit a feature that has not been found in other Lepidoptera. On the dorsum of abdominal segments 7–9 there are two rows of setae, similar in appearance to those on the cremaster. These

PHYLOGENY OF THE JOSIINI

Figure 20. Final instar larvae of Josiini in lateral view, anterior at left. A, Metathorax and A1 of Cyanotricha necyria (50 3 ); B. Anal plate of Josia draconis (97 3 ) showing four setae (Character 79.0); C, Anal plate of Thirmida superba (82 3 ) showing seven setae (Character 79.1); D, Lateral plate of A10 proleg (62 3 ) in Zunachetha annulata (outgroup) with only four setae (Character 80.0); E, A10 proleg of Josia megaera (101 3 ) with nine setae (Character 80.1); F, A10 proleg of Thirmida superba (61 3 ) with more than 15 setae (Character 80.2).

33

34

J. S. MILLER

Figure 21. Pupal cremaster of Josiini (Characters 83–86). A, Getta baetifica (53 3 ), dorsal view; B, Josia striata, ventral view (70 3 ); C, Josia flavissima, dorsal view (57 3 ); D, Josia flavissima (120 3 ) showing cremaster setae; E, Cyanotricha necyria, dorsal view (88 3 ); F, Cyanotricha necyria (200 3 ) showing cremaster setae.

PHYLOGENY OF THE JOSIINI

35

have been described in pupae of Phryganidia (Mosher, 1916; Beutelspacher, 1986; Miller, 1987) and Zunacetha (Miller, 1992a), and they also occur in Tithraustes. Absence of abdominal setae in the pupa (e.g. Fig. 21A) is apparently a synapomorphy for the Josiini (Table 4), but they evolved secondarily in a single josiine species, Cyanotricha necyria (Fig. 21E).

DISCUSSION

Comparative anatomy of the Josiini reveals a remarkable diversity of character evolution within a well-defined clade that has probably undergone relatively recent species radiation. The research described here lays the groundwork for a specieslevel revision of the tribe, and the findings imply that character variation is fully sufficient to facilitate such a study. At least two issues, central to the evolution of josiine moths, deserve further discussion. The first involves the utility of immature stages in elucidating phylogenetic relationships. The second comes from an unexpected finding. Wing pattern characters, which I initially thought could be scored unambiguously, turned out instead to be complex. The intricacies of wing pattern evolution among Josiini, apparent only after cladistic analyses had been completed, provide a classic example of ‘reciprocal illumination’ (Hennig, 1966); once a cladogram has been constructed, reexamination of traits can reveal incorrect hypotheses of homology, and can provide new perspectives on character evolution. The relevant example here is the distinctive longitudinal forewing stripe (LFWS), found in many members of the Josiini. The cladogram built from morphological data (Fig. 3) shows that the LFWS has arisen at least twice within the tribe. A transformation hypothesis of pattern evolution can be proposed which offers intriguing insights into the evolutionary mechanisms that give rise to mimetic associations in Lepidoptera. Comparison of characters from adults and immatures Packard (1895), whose early work on classification of the Notodontidae stands even today as the premiere family treatment, stressed that immature stages are crucial in resolving relationships. His example spurred subsequent notodontid researchers to study larvae in considerable detail (e.g. Forbes, 1948; Godfrey & Appleby, 1987). In accordance with Packard, Miller (1991) found that larval characters were more effective than those from adults in reflecting relationships among subfamilies of the Notodontidae; a phylogeny based on larval traits was better resolved, and the characters themselves showed higher consistency. The explanation for such findings hinges on the theory that larval traits evolve more slowly than those of adults (Alexander, 1990). The preponderance of genitalic characters in adult data sets, and the difficulties involved in homologizing genitalic structures across higher level taxa, may introduce ambiguities not present in larval data (Miller, 1991). To further explore this problem, I partitioned the josiine matrix (Appendix 1) into two data sets: one comprising adult characters only (n = 59) and one for characters of the immature stages (n = 27). These were analyzed separately using character codings identical to the ones employed in the combined analysis. The results are in marked contrast with the subfamily-level studies. For the Josiini, the matrix of adult

36

J. S. MILLER

characters produced only four equally parsimonious trees, and the consensus of those in highly resolved (Fig. 22A). Furthermore, this tree closely matches the cladogram produced by the combined data (Fig. 3). Larval traits were generally less informative (Fig. 22B), and areas of cladogram resolution show poorer topological fit with the tree produced using all data.

Figure 22. Cladograms produced by using either adult or immature characters only. A, Consensus of 4 equally parsimonious trees for adult characters (n = 59), length = 150, CI = 0.65, RI = 0.86; B, Consensus of 231 equally parsimonious trees for immature characters (n = 27), length = 55, CI = 0.61, RI = 0.81.

PHYLOGENY OF THE JOSIINI

37

Two factors probably combine to produce this result. The first seems intuitively obvious; I discovered fewer characters from immatures–approximately half as many. This factor alone may not be a sufficient explanation, however. In the subfamily-level studies (Miller, 1991), larval traits gave a superior result even though adult characters were more numerous. A relatively small data set derived from pupal traits has been similarly effective (Miller, 1992a). Alternatively, if immature characters do in fact evolve more slowly than those of adults, then we may hypothesize that they will be less informative when applied to a recent studies radiation such as the Josiini. Within this tribe, rapidly evolving features such as genitalia seem to be information-rich, and determination of homology is less problematical. Adding credence to this idea is the observation that basal (i.e. older) nodes of the tribal cladogram are accurately resolved by larval data. The observation that adult traits may be sufficient for elucidating relationships in the Josiini can only be cause for celebration. Life history information for Neotropical moth groups is extremely scarce and difficult to acquire. This experiment indicates that relationships among the other 79 described species of Josiini can potentially be resolved using museum-preserved adult material, the only resource currently available.

Wing pattern evolution The incredible diversity of wing colours and patterns among Lepidoptera is perhaps the most intriguing feature of their evolution. Bright coloration can serve as a signal during courtship, as a means of advertising toxicity to potential predators or, in palatable species, to provide protection through mimetic resemblance to a toxic model (Silberglied, 1984; Scoble, 1992; Vane-Wright & Boppr´e, 1993). Although bright coloration has clearly arisen many times within the order (Scoble, 1992), only recently have specific cases been examined using phylogenetic methods. With reference to mimetic patterns, we are now beginning to understand how these evolve, and we can, for the first time, propose transformational hypotheses for future testing. The function of wing colours in Josiini remains open to speculation. Little is known about mating behaviour in the group, and their palatability to potential predators has not been tested. However, since the caterpillars feed on Passiflora as do those of Heliconiini, and since it has been shown that Heliconius species are generally toxic to vertebrates (Jones, 1934; Brower, Brower & Collins, 1963; Brower & Brower, 1964; Brower, 1984; Chai, 1986), one can infer that josiines are poisonous as well. Their brilliant colours would therefore be aposematic. The forewings of species in the Josiini exhibit one of two basic pattern types. In many, the wing is dark (almost black) with a white, yellow or orange transverse band ( = TFWB) (Figs 1, 6A, 6C, 6D, 7A). The other pattern type comprises a dark ground colour with a yellow or orange longitudinal stripe ( = LFWS) of variable length. The hindwing may be either entirely dark, or with a central area of colour of various shapes and sizes (Fig. 1). In species with a LFWS, the hindwing frequently has an elongate longitudinal stripe (LHWS) matching the one in the forewing (Fig. 7C, D). A few taxa, such as Thirmida (Fig. 1), have markings that do not easily fit into these categories. Josia megaera (Figs. 1, 6A) is unique; it is the only josiine in which the

38

J. S. MILLER

forewing exhibits both transverse and longitudinal pattern elements, and there is an unusual arrangement of orange regions in the hindwing as well. In nymphalid butterflies, pattern elements are almost always black or dark brown while light-coloured regions constitute the background (Higgins, 1868; Nijhout et al., 1990). In nymphalids with wings that are largely or completely black, the pattern is formed by expansion and fusion of elements (Nijhout & Wray, 1988). It is hard to imagine such a system at work in the Josiini. Pattern expression in these moths may differ fundamentally from that of butterflies. In the absence of genetic or developmental data, it seems most parsimonious to assume that the light regions on the wing in Josiini are pattern elements while melanic areas are background. A casual survey of pattern types across diurnal Lepidoptera suggests that the TFWB is common. It occurs in some form or other in members of almost all butterfly subfamilies, and is frequent in diurnal moths such as the Arctiidae (Fig. 1). In contrast, the LFWS is rare (Seitz, 1916). In species where it does occur, the resemblance to Josiini is remarkable. Neotropical Lepidoptera with orange longitudinal wing stripes against a dark background (Fig. 1) include species of Chrysophila H¨ubner (Pyralidae), Mesenopsis Godman & Salvin (Riodinidae), and Josiomorpha Felder (Arctiidae). Whether these taxa, together with Josiini, form Batesian or M¨ullerian mimicry complexes is open to speculation. Palatability tests are sorely needed. According to the josiine phylogeny, which must be considered robust given the range of morphological characters upon which it is based, the LFWS evolved twice within the tribe: once in Clade 3, and again in Clade 21 (Fig. 23). The LHWS shows a similar distribution. When wing pattern characters are completely removed from the analysis, the same cladogram results. Careful inspection of the trait I initially defined as ‘forewing with a longitudinal stripe from base’ (Character 13) reveals that the location of the stripe differs in these two clades. In species of Clade 13, exemplified by Josia insincera (Fig. 6B) and J. frigida (Fig. 7D), the stripe straddles the cubital vein as it passes through the discal cell, and it crosses the fork of M3 + CuA1 at its distal end. The anterior margin of the stripe in these species never extends forward into the discal cell as far as the radius. The forewing stripe in members of Clade 21 is distinctly different. Here, it is located more anteriorly. In Josia cruciata (Fig. 7C), for example, the anterior margin of the stripe touches the subcosta near its base, and the stripe passes almost through the middle of the discal cell, touching the radius. In members of Clade 21, the LFWS never straddles the cubital vein or the fork of M3 + CuA1. The hindwing stripe shows analogous differences between clades. There are also two types of transverse bands. The TFWB in most species is located beyond the discal cell (Figs 6A, 6C, 7A). Alternatively, in the taxa of Clade 17 where a TFWB occurs, such as J. ena (Fig. 6D), the band is located across the end of the discal cell. The first of these character states is assumed to be primitive, since it is exhibited by J. megaera (Fig. 6A). In summary, wing pattern evolution within the Josiini is more complex than it first appears. A classic criterion of homology is similarity of placement (Remane, 1952; Hennig, 1966). Josiine wings exhibit clear differences in the position of pattern elements. In conjunction with the other character evidence, these findings demonstrate that the longitudinal forewing and hindwing stripes of the two josiine clades are not homologous. M¨ullerian mimicry, defined as convergent evolution of

PHYLOGENY OF THE JOSIINI

39

the same signalling system in multiple aposematic species (M¨uller, 1879; Brower, 1995), is thus revealed by phylogenetic analysis. There has been considerable discussion concerning the lability of mimetic patterns in Lepidoptera. M¨ullerian mimics arise within clades that are already aposematic. Since, in such cases, the fitness of fine-tuned resemblance is positively density dependent, it has been argued on theoretical grounds that M¨ullerian mimicry can evolve rapidly and repeatedly (Turner, Kearney & Exton, 1984; Mallet & Singer,

Figure 23. Cladogram of relationships among Josiini (from Fig. 3) with wing patterns of representative terminal taxa overlaid, showing convergent evolution of the longitudinal FW stripe. Species shown (from top to bottom) are: Josia megaera, J. frigida, J. aurifusa, J. insincera, Thirmida superba, Getta baetifica, Josia fornax, J. fluonia, J. annulata and J. striata

40

J. S. MILLER

1987; Brower, 1994b). A mechanism for such rapid change has been proposed. In co-mimicking species of Heliconius, pattern expression apparently involves the same pattern elements and largely the same genes (Sheppard et al., 1985; Nijhout, 1991). The genetic equipment needed to produce similarity thus exists in all species within the genus. The emerging view of pattern evolution in butterflies suggests that “dramatic phenotype changes are brought about primarily by small changes in the sizes of pattern elements or small shifts in their positions” (Nijhout, 1994a: 153). These arguments can be applied to wing pattern evolution among Josiini. Assuming that all species arose from an aposematic common ancestor (associated with either Passiflora or Turnera), the theoretical arguments of Turner et al. (1984) and Mallet & Singer (1987) are upheld; a rare phenotype (the LFWS) has evolved twice within an aposematic clade. By overlaying wing pattern characters onto the josiine cladogram (Fig. 23), separate transformation hypotheses, leading to the evolution of LFWS’s in Clades 3 and 21, can be inferred. First, elongation of the short basal stripe of Josia megaera, perhaps resulting from fusion with the transverse band (Fig. 6A), could produce a LFWS similar in position to the ones found in Clade 3 (Figs 6B, 7D). A second evolutionary scenario would entail transformation of the transverse band in J. flavissima and J. ena (Fig. 6D) into the LFWS of species in Clade 21 (Fig. 7C), with the pattern of J. fluonia (Fig. 7B) being an intermediate character state. Having pointed to the rarity of longitudinal wing stripes in Lepidoptera, it is interesting to note the unique occurrence of them in a dioptine genus unrelated to the Josiini. Erbessa mimica Hering is known from a single specimen, captured among a series of Josia oribia Druce with which it is an almost perfect mimic, along the Rio Songo of Bolivia (Hering, 1925). The 66 species of Erbessa exhibit an amazing diversity of aposematic wing patterns (Hering, 1925), but none of these is even roughly similar to the pattern of E. mimica. What may represent another case of rapid mimetic convergence must await testing through phylogenetic analyses of Erbessa. Classification of the Josiini The cladogram shows that Josia, as previously conceived, is a polyphyletic genus. The monophyly of other josiine genera is suspect. For example, Thirmida may be paraphyletic with respect to Cyanotricha (Fig. 3, Clade 12). Such results are not surprising since none of these taxa have been revised, and our present day concepts are largely based on the ideas of taxonomists working before the turn of the century. Most early workers relied exclusively on wing patterns for arranging species into genera, and the rampant mimicry in Dioptinae confounded their attempts to build a stable classification (Seitz, 1925). Before this paper, the only other traits used in classifying Josiini were the branching patterns of the wing veins (Prout, 1918), a character complex notorious for its high levels of homoplasy (e.g. Janse, 1920; Weller, 1989). However, these are not meant to be criticisms of the contributions of early taxonomists. As systematics proceeds, we develop more and more precise estimates of phylogenetic relationships. These in turn will ultimately lead to stable classifications (Wheeler, 1990). Although my cladograms point out a dire need for revised generic concepts in the Josiini, I make no formal nomenclatural changes here. Generic names exist for many Josia subclades. For example, the name Lyces Walker could be applied to the species

PHYLOGENY OF THE JOSIINI

41

of Clade 17. Genitalic dissections reveal that the type species of Lyces, angulosa Walker, is closely related to fornax and flavissima (J.S. Miller, unpubl. data). However, I refrain from such proposals until a revision of the tribe has been undertaken. Comprehensive study of josiine relationships, a goal of future research, will likely result in a very different generic classification from the one in use today.

ACKNOWLEDGEMENTS

Field work forms the core of this paper. I am especially grateful to Cal Snyder (AMNH) and L.D. Otero (M´erida, Venezuela), both of whom have lent their vast expertise to helping discover josiine life histories. This paper would not have been possible without their tireless efforts. It is also a pleasure to acknowledge the expert field assistance of A. Aiello (Balboa, Panam´a) and E. Tapia (Quito, Ecuador). Many people have offered unlimited hospitality, logistical support and friendship during field trips. I am grateful to G. Onore (Quito, Ecuador), B. Hermier, F. Beneluz and J.J. de Granville (French Guiana), V. Savini, L. Joly and J. Clavijo (Venezuela), J. Corales and I. Chac´on (Costa Rica), and to Don Windsor (Panam´a). For the loan of larval and adult museum material, I thank J.E. Rawlins (Carnegie Museum of Natural History), M. Honey and I. Kitching (The Natural History Museum, London), W. Mey (Zoologisches Museum, Humboldt-Universit¨at, Berlin), B. Poole (United States National Museum) and R. Friesen (USDA, Hilo, Hawaii). The genitalia drawings in Figures 10, 11, 13, 14A, 14B and 15 were done by Amy Trabka. For assistance with scanning electron microscopy I thank P. Phong-Melville and W. Barnett of the AMNH Interdepartmental Laboratories. I also wish to extend my sincere gratitude to the people at the various agencies who have issued collecting permits, and who have graciously allowed me and my colleagues to collect in their countries. In Costa Rica I would like to thank Maria Luisa Alfaro of the Ministerio de Recursos Naturales Energia y Minas, Juan Miguel Sanchez of the servicio de Parques Nacionales, and Juan Diego Alfaro Fernandez of La Amistad. In Panam´a, I am grateful to the Smithsonian Tropical Research Institute (STRI), the Instituto de Recursos Hidr´aulicos y Electrificaci´on (IRHE), the Instituto de Recursos Naturales Renovables (INRENARE), and the Proyecto de Estudio para el Manejo de Areas Silvestres de Kuna Yala (PEMASKY). For permission to study in Venezuela, my thanks go to the Instituto Nacional de Parques (INPARQUES) and to Alberto Fern´andez Badillo, Director of the Estaci´on Biol´ogica Dr Alberto Fern´andez Y´epez at Rancho Grande. In Ecuador, I thank Dr Sergio Figueroa S., Director de Areas Naturales, Ministerio de Agricultura y Ganaderia. For comments on an earlier draft of this paper and for discussions of mimicry, I thank A.V.Z. Brower and C. Snyder. I also thank two anonymous reviewers. This research was funded by National Science Foundation Grant no. BSR-9106517.

REFERENCES Ackery PR. 1988. Hostplants and classification: a review of nymphalid butterflies. Biological Journal of the Linnean Society 33: 95–203. Alexander B. 1990. A cladistic analysis of the nomadine bees (Hymenoptera: Apoidea). Systematic Entomology 15: 121–152.

42

J. S. MILLER

Barth R. 1955. Maennliche Duftorgane brasilianischer Lepidopteran. 10. Mitteilung: Hemiceras proximata Dogn. (Notodontidae). Anais da Academia Brasileira de Ciˆencias 27: 539–544. Bates HW. 1862. Contributions to an insect fauna of the Amazon Valley. Lepidoptera: Heliconidae. Transactions of the Linnean Society of London 23: 495–566. Benson WW, Brown KS, Gilbert LE. 1976. Coevolution of plants and herbivores: passion flower butterflies. Evolution 29: 659–680. Beutelspacher CR. 1986. Una especie nueva Mexicana de la familia Dioptidae de importancia forestal (Lepidoptera: Dioptidae). Anales del Instituto de Biologia de la Universidad National Aut´onoma de Mexico 56, Serie Zoolog´ıa 2: 477–482. Brower AVZ. 1994a. Phylogeny of Heliconius butterflies inferred from mitochondrial DNA sequences. Molecular Phylogeny and Evolution 3: 159–174. Brower AVZ. 1994b. Rapid morphological radiation and convergence among races of the butterfly Heliconius erato inferred from patterns of mitochondrial DNA evolution. Proceedings of the National Academy of Sciences, USA 91: 6491–6495. Brower AVZ. 1995. Locomotor mimicry in butterflies? A critical review of the evidence. Philosophical Transactions of the Royal Society of London, Series B. 347: 413–425. Brower AVZ. 1996. Parallel race formation and the evolution of mimicry in Heliconius butterflies: a phylogenetic hypothesis from mitochondrial DNA sequences. Evolution 50: 195–221. Brower LP. 1984. Chemical defense in butterflies. In: Vane-Wright RI, Ackery PR, eds. The biology of butterflies. Royal Entomological Society of London 11: 109–134. London: Academic Press. Brower LP, Brower JVZ. 1964. Birds, butterflies, and plant poisons: a study in ecological chemistry. Zoologica (New York) 49: 137–159. Brower LP, Brower JVZ, Collins CT. 1963. Experimental studies of mimicry. 7. Relative palatability and M¨ullerian mimicry among Neotropical butterflies of the subfamily Heliconiinae. Zoologica (New York) 48: 65–83. Bryk F. 1930. Dioptidae. In: Strand E, ed. Lepidopterorum Catalogus (42). Berlin: W. Junk. Carroll SB, Gates J, Keys DN, Paddock SW, Panganiban GEF, Selegue JE, Williams JA. 1994. Pattern formation and eyespot determination in butterfly wings. Science 265: 109–114. Cavin JC, Bradley TJ. 1988. Adaptation to ingestion of beta-carboline alkaloids by Heliconiini butterflies. Journal of Insect Physiology 34: 1071–1075. Chai P. 1986. Field observations and feeding experiments on the responses of rufous-tailed jacamars (Galbula ruficauda) to free-flying butterflies in a tropical rainforest. Biological Journal of the Linnean Society 29: 161–189. Common IFB. 1990. Moths of Australia. Carlton, Victoria: Melbourne University Press. Croat TB. 1978. Flora of Barro Colorado Island. Stanford, California: Stanford University Press. Cronquist A. 1988. The Evolution and Classification of Flowering Plants, Second Edition. Bronx, NY: New York Botanical Garden. DeVries PJ. 1987. The Butterflies of Costa Rica and their Natural History. Papilionidae, Pieridae, Nymphalidae. Princeton: Princeton University Press. Ehrlich PR. 1958. The integumental anatomy of the monarch butterfly Danaus plexippus L. (Lepidoptera: Danaidae). University of Kansas Science Bulletin 38: 1315–1349. Ehrlich PR, Raven PH. 1964. Butterflies and plants: a study in coevolution. Evolution 18: 586–608. Farris JS. 1988. Hennig86 reference. Version 1.5. Published by the author. Ferguson DC. 1985. Contributions toward reclassification of the world genera of the tribe Arctiini, Part 1: Introduction and a revision of the Neoarctia-Grammia group (Lepidoptera: Arctiidae; Arctiinae). Entomography 3: 181–275. Forbes WTM. 1922. Miscellaneous notes. Journal of the New York Entomological Society 30: 71–72. Forbes WTM. 1931. Notes on the Dioptidae (Lepidoptera). Journal of the New York Entomological Society 39: 69–76. Forbes WTM. 1939. Family Dioptidae, The Lepidoptera of Barro Colorado Island, Panama. Bulletin of the Museum of Comparative Zoology 85: 99–322. Forbes WTM. 1948. Lepidoptera of New York and neighbouring states. Part 2. Notodontidae. Cornell University Agricultural Experiment Station Memoirs 274: 203–237. Fracker SB. 1915. The classification of lepidopterous larvae. Illinois Biological Monographs 2: 1–161. Franclemont JG. 1946. A revision of the species of Symmerista H¨ubner known to occur north of the Mexican border (Lepidoptera, Notodontidae). Canadian Entomologist 78: 96–103. Fullard JH. 1984. External auditory structures in two species of Neotropical notodontid moths. Journal of Comparative Physiology 155: 625–632. Gentry AH. 1993. A Field Guide to the Families and Genera of Woody Plants of Northwest South America (Colombia, Ecuador, Peru) With Supplementary Notes on Herbaceous Taxa. Washington, DC: Conservation International, 895 pp. Godfrey GL, Appleby JE. 1987. Notodontidae (Noctuoidea): the Notodontids and Prominents. In: Stehr FW, ed. Immature Insects. Dubuque, Iowa: Kendall/Hunt, 523–533. Harvey DJ. 1991. Higher classification of the Nymphalidae. Appendix B In: Nijhout HF, ed. The Development and Evolution of Butterfly Wing Patterns. Washington, DC: Smithsonian Press, 255–268. Hennig W. 1966. Phylogenetic Systematics. Urbana: University of Illinois Press.

PHYLOGENY OF THE JOSIINI

43

Hering EM. 1925. Dioptidae. In: Seitz A, ed. Macrolepidoptera of the World, Vol. 6. Stuttgart: Alfred Kernen, 501–534. Heywood VH. 1979. Flowering Plants of the World. London: Oxford University Press. Higgins HH. 1868. On the colour-patterns of butterflies. Quarterly Journal of Science 5: 323–329. Hinton HE. 1946. On the homology and nomenclature of the setae of lepidopterous larvae, with some notes on the phylogeny of the Lepidoptera. Transactions of the Royal Entomological Society of London 97: 1–37. Holm-Nielsen LB, Jorgensen PM, Lawesson JE. 1988. 126. Passifloraceae. In: Harling G, Andersson L, eds. Flora of Ecuador, No. 31. Janse AJT. 1920. On the South African Notodontidae, with descriptions of apparently new genera and species. Annals of the Transvaal Museum 7: 149–237. Janzen D. 1983. Erblichia adorata Seem. (Turneraceae) is a larval host plant of Eueides procula vulgiformis Butler and Druce (Nymphalidae-Heliconiini) in Santa Rosa National Park. Journal of the Lepidopterists’ Society 37: 70–78. Jones FM. 1934. Further experiments on colouration and relative acceptability of insects to birds. Transactions of the Royal Entomological Society of London 82: 443–453. Killip EP. 1938. The American species of Passifloraceae. Publications of the Field Museum of Natural History (Botanical Series) 19: 1–613. Kiriakoff SG. 1950. Recherches sur les organes tympanique des L´epidopt`eres en rapport avec la classification. III. Dioptidae. Bulletin et Annales de la Societ´e Entomologique de Belgique 86: 67–86. Kitching IJ. 1984. The use of larval chaetotaxy in butterfly systematics, with special reference to the Danaini (Lepidoptera: Nymphalidae). Systematic Entomology 9: 49–61. Kohler ¨ P. 1930. Los Dioptidae argentinos. Revista de la Sociedad Entomologia de Argentina 14: 153–162. Mallet J, Singer MC. 1987. Individual selection, kin selection, and the shifting balance in the evolution of warning colours: the evidence from butterflies. Biological Journal of the Linnean Society 32: 337–350. Miles DB, Dunham AE. 1993. Historical perspectives in ecology and evolutionary biology: the use of phylogenetic comparative analysis. Annual Review of Ecology and Systematics 24: 587–619. Miller JS. 1987. A revision of the genus Phryganidia Packard, with description of a new species (Lepidoptera: Dioptidae). Proceedings of the Entomological Society of Washington 89: 303–321. Miller JS. 1988. External genitalic morphology and copulatory mechanism of Cyanotricha necyria (Felder) (Noctuoidea: Dioptidae). Journal of the Lepidopterists’ Society 42: 103–115. Miller JS. 1989. Euchontha Walker and Pareuchontha new genus (Lepidoptera: Dioptidae): a revision, including description of three new species, and discussion of a male forewing modification. American Museum Novitates 2938: 1–41. Miller JS. 1991. Cladistics and classification of the Notodontidae (Lepidoptera: Noctuoidea) based on larval and adult morphology. Bulletin of the American Museum of Natural History 204: 230 pp. Miller JS. 1992a. Pupal morphology and the subfamily classification of the Notodontidae (Lepidoptera: Noctuoidea). Journal of the New York Entomological Society 100: 228–256. Miller JS. 1992b. Host-plant associations among prominent moths. BioScience 42: 50–57. Miller JS, Otero LD. 1994. Immature stages of Venezuelan Dioptinae (Notodontidae) in Josia and Thirmida. Journal of the Lepidopterists’ Society 48: 338–372. Miller JS, Wenzel JW. 1995. Ecological characters and phylogeny. Annual Review of Entomology 40: 389–415. Mosher E. 1916. A classification of the Lepidoptera based on characters of the pupa. Bulletin of the Illinois State Laboratory of Natural History 12: 14–159. Muller ¨ F. 1879. Ituna and Thyridia: a remarkable case of mimicry in butterflies. Proceedings of the Royal Entomological Society of London 1879: xx-xxix. Nahrstedt A, Davis RH. 1981. The occurrence of cyanoglucosides, linamarin and lotaustralin, in Acraea and Heliconius butterflies. Comparative Biochemistry and Physiology 68(B): 575–577. Nahrstedt A, Davis RH. 1983. Occurrence, variation and biosynthesis of the cyanogenic glucosides linamarin and lotaustralin in species of the Heliconiini. Comparative Biochemistry and Physiology 75(B): 65–73. Nielsen ES, Common IFB. 1991. Lepidoptera (Moths and butterflies), pp. 817–915. In: The Insects of Australia, Second Edition, Vol. II. Victoria, Australia: Melbourne University Press (CSIRO). Nijhout HF. 1990. A comprehensive model for colour pattern formation in butterflies. Proceedings of the Royal Society of London 239: 81–113. Nijhout HF. 1991. The Development and Evolution of Butterfly Wing Patterns. Washington, DC: Smithsonian Institution Press, 297 pp. Nijhout HF. 1994a. Developmental perspectives on evolution of butterfly mimicry. BioScience 44: 148–157. Nijhout HF. 1994b. Genes on the wing. Science 265: 44–45. Nijhout HF, Wray GA. 1986. Homologies in the colour patterns of the genus Charaxes (Lepidoptera: Nymphalidae). Biological Journal of the Linnean Society 28: 387–410. Nijhout HF, Wray GA. 1988. Homologies in the colour patterns of the genus Heliconius (Lepidoptera: Nymphalidae). Biological Journal of the Linnean Society 83: 345–365. Nijhout HF, Wray GA, Gilbert LE. 1990. An analysis of the phenotypic effects of certain colour pattern genes in Heliconius (Lepidoptera: Nymphalidae). Biological Journal of the Linnean Society 40: 357–372. Oseto CY, Helms TJ. 1976. Anatomy of the adults of Loxigrotis albicosta. University of Nebraska Studies: New series 52: 1–127. Lincoln: University of Nebraska.

44

J. S. MILLER

Packard AS. 1895. Monograph of the bombycine moths of America north of Mexico including their transformations and origin of the larval markings and armature, part I: family 1, Notodontidae. Memoirs of the National Academy of Sciences 7: 1–390. Peterson A. 1962. Larvae of Insects, Part I: Lepidoptera and Plant Infesting Hymenoptera. Columbus, Ohio: Edwards Brothers, 315 pp. Powell JA. 1973. A systematic monograph of the New World ethmiid moths (Lepidoptera: Gelechioidea). Smithsonian Contributions to Zoology 120: 1–302. Prout LB. 1918. A provisional arrangement of the Dioptidae. Novitates Zoologicae 25: 395–429. Richards AG. 1932. Comparative skeletal morphology of the noctuid tympanum. Entomologica Americana 13: 1–43. Remane A. 1952. Die Grundlagen des Nat¨urlichen Systems der Vergleichenden Anatomie und der Phylogenetik. Leipzig: Geest und Portig KG. Scoble MJ. 1992. The Lepidoptera: Form, Function and Diversity. New York: Oxford University Press, 404 pp. Seitz A. 1916. Family: Erycinidae. In: Seitz A, ed. Macrolepidoptera of the World, Vol. 5. Stuttgart: Alfred Kernen, 617–738. Seitz A. 1925. Dioptidae, general topics. In: Seitz A, ed. Macrolepidoptera of the World, Vol. 6. Stuttgart: Alfred Kernen, 499–500. Sheppard PM, Turner JRG, Brown KS, Benson WW, Singer MC. 1985. Genetics and the evolution of Muellerian mimicry in Heliconius butterflies. Philosophical Transactions of the Royal Society of London, B 308: 433–613. Sick H. 1940. Beitrag zur Kenntnis der Dioptidae, Notodontidae und Thaumetopoeidae und deren Verwandtschaftsbeziehungen zueinander. Zoologische Jahrbucher. Abteilung fur Anatomie und Ontogenie de Tiere 66: 263–290. Silberglied RE. 1984. Visual communication and sexual selection among butterflies. In: Vane-Wright RI, Ackery PR, eds. The biology of butterflies. Royal Entomological Society of London 11: 207–223. London: Academic Press. Spencer KC. 1988. Chemical mediation of coevolution in the Passiflora-Heliconius interaction. In: Spencer KC, ed. Chemical mediation of coevolution. New York: Academic Press, 167–240. Stehr FW. 1987. Order Lepidoptera. In: Stehr FW, ed. Immature insects. Dubuque, Iowa: Kendall/Hunt, 288–305. Swofford D. 1991. Phylogenetic analysis using parsimony. Version 3.0S. Published by the author. Todd EL. 1973. Taxonomic and distributional notes on some species of Nystalea Guen´ee, with special emphasis on the species of the continental United States (Lepidoptera: Notodontidae). Proceedings of the Washington Entomological Society 75: 265–275. Turner JRG, Kearney EP, Exton LS. 1984. Mimicry and the Monte Carlo predator: the palatability spectrum and the origins of mimicry. Biological Journal of the Linnean Society 23: 247–268. Vane-Wright RI, Boppr´e M. 1993. Visual and chemical signalling in butterflies: functional and phylogenetic perspectives. Philosophical Transactions of the Royal Society of London (Series B) 340: 197–205. Vane-Wright RI, Smith CR. 1991. Phylogenetic relationships of three African swallowtail butterflies, Papilio dardanus, P. phorcas and P. constantinus: a cladistic analysis (Lepidoptera: Papilionidae). Systematic Entomology 16: 275–291. Weller SJ. 1989. Phylogeny of the Nystaleini (Lepidoptera: Noctuoidea: Notodontidae). Unpublished Ph.D. thesis. University of Texas, Austin. Weller SJ. 1990. Revision of the Nystalea aequipars Walker species complex with notes on nystaleine genitalia (Lepidoptera: Notodontidae). Journal of the New York Entomological Society 98: 35–49. Weller SJ. 1991. Revision of the Pentobesa xylinoides (Walker) species group (Lepidoptera: Notodontidae). Proceedings of the Entomological Society of Washington 93: 795–807. Wheeler QD. 1990. Insect diversity and cladistic constraints. Annals of the Entomological Society of America 83: 1031–1047. Wolda H, Foster R. 1978. Zunacetha annulata (Lepidoptera: Dioptidae), an outbreak insect in a Neotropical forest. Geo-Eco-Trop 2: 443–454.

Zunacetha Tithraustes C. necyria G. baetifica J. annulata J. auriflua J. aurifusa J. cruciata J. draconis J. ena J. interrupta J. flavissima J. fluonia J. fornax J. frigida J. gigantea J. gopala J. insincera J. ligata J. megaera J. radians J. striata J. turgida P. fasciculosa T. discinota T. superba

20 00100001100000000000 01000001100000000000 10210010000010021020 22111011010100021020 02010010000020121110 00010010000020121010 00010010000021121010 02010010000020121110 02011010000110021010 02010010000200021020 10110010000020021010 02010110000200021020 02010010000210021020 02010010000200021020 00010030001020121010 00010020001020121011 02010110000020021030 10110010000020021010 00010020001020221011 01110010000111010010 00010020001020221011 02010010000020021030 00010010000021121010 22111011010100021020 01210010000010021020 01210010000010021020

40 00020000000000000000 00000000000000000000 30001020110001002000 20021120200000012100 10011021220022022110 12001010002000001001 12001010002000001001 10011021220022022110 10001020210000002100 40021011210010102110 10101010001100001002 10001021210010102110 11011021210002022110 11001021210000022110 42001010001000011001 42001010001000011001 10012021220022022110 40101010001100001002 42001010001000011001 10001010000000001001 42001010001000011001 11012021220012022110 42001010002000001001 300211202000000?2100 30001020110001002000 30001020110001002000

60 00000000000000000000 00000000000000000000 00010010011010001012 01012040001000000001 00111030111001100121 00020001010112200001 00020001010112200001 00111030111001100121 010120000110110000?1 10210020012001000121 00010020010102010001 10210023012001100121 00111030111021100121 00110031111021100121 00020000011110200001 00020000011111200001 00111032010021100121 00010020010102010001 00020101011111200001 00010003010111000000 00020101011111200001 00111032010021100121 00020001010112200001 01012040001000000001 00010010011010001012 00010011011010001011

80 00000000000000000000 00000000000000000000 11110002000013000112 11100100100004010001 11110000010011010000 11110001002001110001 11110001002001110001 11110010010011010000 11100100010004010001 11100010000014010000 11110001001001110001 11100001000011010000 11100000010011010000 11100011010011010000 11100100001001110001 11100001001002110001 11110000010011011000 11110001001001110001 11100001001102110001 10100000010001010001 11100001001102110001 11110000010011011000 11110001002001110001 11100110100004010002 11101002000013000112 11101002000013010012

000001 0?0001 111000 000010 000010 000010 000010 000010 010010 000010 000010 010100 000110 000110 000010 000010 000100 000010 000010 000010 000010 000100 000010 000010 111010 011010

Appendix 1. Matrix of characters for the Josiini and two outgroup taxa (Zunacetha and Tithraustes). Number of characters=86, number of taxa=26. Analysis yielded the cladogram in Fig. 3

PHYLOGENY OF THE JOSIINI 45