Peru, Arequipa, Pacaychacra [15°54'S, 72°33'W], 1500 m.

Holotype ♂, Peru, Arequipa, Pacaychacra, 15°54'S, 72°33'W, 1500 m, reared from eggs collected on Malesherbia tenuifolia (Passifloraceae), 24.VII.2019, J. Farfán leg. deposited in MUSM. Paratypes (25♂, 17♀): Peru. Lima: 1♂, San Bartolomé, 1600 m, [11°55'S, 76°31'W], 21.iii.[19]81, P. Hocking (MUSM); 1♀, Cocachacra, 1450 m, [11°55'S, 76°32'W], 6.x.[19]83, P. Hocking [MUSM-ENT 008630] (MUSM); 1♀, Río Rímac, Chaute, 2350 m, 11°56'S, 76°30'W, 12.v.2012, P. Hocking (MUSM). Arequipa: 2♀, 7 km E Cháparra, 1450m, [15°41'S, 73°49'W], 14.iv.[19]88, G. Lamas [MUSM-ENT 008631, 008632] (MUSM); 1♂, entre Majes y Chuquibamba, 15°55'S, 72°33'W, 1500 m, 24.iv.2017, G. Lamas (MUSM); 1♂ same data as holotype [LMCI 357–51] (LMCI); 1♂ same data as holotype (MUSA); 1♂, 2♀, same data as holotype but with date 15.XII.2020 (MUSA); 1♂ same data as holotype but with date 24.vii.2021 (MUSA); 4♂ Aplao, Valle Majes, 15°53'46"S, 72°28'03"W, 800 m, 02.vi.2013, Leg. J. Cerdeña / M. Delgado (MUSA); 1♂, Condesuyos, Pacaychacra,15°54'59.2"S, 72°33'01.5"W, 1500 m, 02.IX.2020, Leg. Jose Cerdeña (MUSA); 1♂, same data, but 13.XII.2020 (MUSA); 1♀, same data, but 24.VII.2021, Jackie Farfán (MUSA); 1♀, same data, but 12.XI.2019, [LMCI 357–52] (LMCI); 3♂, Yura, 2 Km SW Yura viejo, 16°13'19"S, 71°42'23"W, 2600 m 21.IV.2022, Leg. Jackie Farfán (MUSA). Moquegua: 6♂, 1♀, Torata, 170630/705036 [17°06'30"S, 70°50'36"W], 2090m, 26.VII.2021, Leg. Jackie Farfán (MUSA); 1♂, 1♀, same data [LMCI 357–53, LMCI 357–54] (LMCI); 4♀, La Capilla, 13 Km S Puquina, 164450/711054 [16°44'50"S, 71°10'54"W], 1800 m, 28.XII.2013, Leg. J. Farfán / J. Cerdeña (MUSA); 1♀, Omate, 3 Km SW Omate, 16°41'15.27"S, 70°59'13.73"W, 2000 m, 27.X.2017, Leg. Robert Cornejo (MUSA). Tacna: 1♂, Chululuni, 17°22'02"S, 70°28'24"W, 1800 m, 18.XII.2020, Leg. Jackie Farfán (MUSA). CHILE. Arica: 1♂, Las Peñas, 18°33'08"S, 69°46'03"W, 1580 m, 02.XII.2020, H. A. Vargas leg. [IDEA-LEPI–2022–008] (IDEA); 1♀, same data, but 10.XI.2020; [IDEA-LEPI–2022–007] (IDEA). Tarapacá: 1♀, Iquique, Parca, 3000 m, [20°01'S, 69°01'W], ix/x.[19]51, L. E. Peña (MUSM); 1♂, Q[uebrada] de Guatacondo, “Cauquenisca” [= Cautenicsa], 2300 m, [20°56'S, 69°01'W], 26-X-1968, P. Millas (MHNS).

Immature stages preserved in 70% ethanol, with the same data as the holotype, collected on Malesherbia tenuifolia (Passifloraceae) with dates VI.2018, X.2019 and IV.2021, were deposited in LMCI, under accession numbers 357–31 (14 eggs), 357–33, 357–40, 357–44 (20 larvae), 357–46, 357–47, 357–48, 357–49, 357–50 (5 pupae).

Dione (Agraulis) dodona sp. nov. can be easily distinguished from its congeners by the wing pattern, presenting a black postdiscal spot between M₃-A₁ veins on the dorsal hindwing that is absent in all other species, and, also, by presenting a divided or partially divided silver spot in the discal cell on the ventral hindwing, always undivided in other species of Dione (Agraulis). In the male genitalia, the valvae have a rounded termen, without denticles, and the distal portion of the crista is narrow and straight. In other species, the termen is sub-triangular and shows denticles on the margin, and the distal apex of the crista is transversally enlarged. Also, the juxta has its upper edge slightly split in dodona, which is widely open in other species. The aedeagus is straight in lateral view without cornuti in dodona, up-curved near distal end with cornuti in other species. The female genitalia possess evenly wide signa in the proximal portion, composed of robust spines, unlike other species that have smaller spines; the proximal apex of signa is narrower and progressively enlarges distally.

Furthermore, the immature stages of Dione (Agraulis) dodona sp. nov. show differences with the available data compared to other Agraulis species (Beebe et al. 1960; Brown 1981; da Silva et al. 2006). In the egg, the number of horizontal carinae almost doubles the number described for Dione (Agraulis) maculosa [cited as ‘A. vanillae´] (11–13 vs. 17–19), the egg being taller in Dione (Agraulis) dodona sp. nov. In relation to the larval stage, in the first instar, the main difference is the size of the D2 setae in the abdominal segments, being very small in maculosa and insularis (1/4 the length of D1) (Beebe et al. 1960; da Silva et al. 2006), but reaching more than half the length of D1 in Dione (Agraulis) dodona sp. nov. (Fig. 9B). In the fifth instar, the head scoli in maculosa are well developed, whereas Dione (Agraulis) dodona sp. nov. bears short stout scoli (Figs 5D, 6E). Furthermore, the prothoracic dorsal plate in Dione (Agraulis) dodona sp. nov. has spine-like setae on top of enlarged conical projection (Fig. 9C), unlike maculosa, which has a simple seta bearing on a small projection (da Silva et al. 2006). In the pupa, the main differences are related to the head protuberances, which are small in Dione (Agraulis) dodona sp. nov. (Fig. 9D), more conspicuous, and as long as half the length of head in maculosa; also, the meso-dorsal crest is less pronounced in Dione (Agraulis) dodona sp. nov. (Fig. 9E), in maculosa the dorsal margins are more enlarged; protuberances on the abdominal segments do not occupy the entire length of the segment in Dione (Agraulis) dodona sp. nov. (Fig. 9F), in contrast to maculosa where protuberances fill all the length of the third abdominal segment.

Adult. (Figs 1–4). Male: Wingspan 44–52 mm (holotype 50 mm). Head: antennae approximately two thirds of the forewing in length, black with the tip of club orange (Fig. 2A), with 36 antennomers, 11 of which define the club. Palpus elongated, approximately twice the size of the head, with a dark brown dorsal color with light orange hairs, in ventral view white covered by white and orange hairs (Fig. 2A). Thorax: Generally brown. Body dorsally black with brown and orange hairs, ventrally covered by white and light orange scales, legs dorsally light orange with white and orange hairs at the base, ventrally white. Forewing length 25–28 mm (holotype: 27 mm), hindwing length 18–20 mm. Wing venation as described in Michener (1942) (Fig. 2B). Wing color pattern typical for the Agraulis clade except a black postdiscal spot between veins M₃-A₁ on dorsal hindwing and a silver spot located in the discal cell on ventral hindwing divided or partially divided (Fig. 1E, AF). Abdomen: dorsally brown with orange hairs, ventrally covered by white scales.

Figure 1. 

Adults of Dione (Agraulis) dodona sp. nov. A, B male, holotype C, D female paratype E, F details of hindwing pattern color (indicated by blue squares in A and D respectively), open arrow points black spot on Cu₁-Cu₂ cell, and close arrow on silver discal spot. Left: dorsal view; right: ventral view. Scale bars: 2 cm.

Figure 2. 

Adult morphology of Dione (Agraulis) dodona sp. nov. A head, male, lateral view B wing venation, male C detail of forewing venation, female (red setae points differences with male) D, E distal portion of prothoracic tarsi under scanning electron microscopy, male in ventral view D female in lateral view E distal tarsomere indicated by open arrow in E. Scale bars: 1 mm (A); 5 mm (B); 50 µm (D); 250 µm (E).

Male genitalia : Rounded and subtriangular valvae occupying most of the genital capsule, being wide anteriorly and narrowest in the apex, with rounded pointed apex with hairs on ventral margin (Fig. 3D), costa with eversible pouch on its inner surface, bearing a median-ventral crista (Fig. 3C); crista narrow, turbinate-shaped, with apex protruded to dorsal margin of valve, and with little spines on surface (Fig. 3B), with wider prolongation on the proximal ventral surface connecting with saccus (Fig. 3A). Saccus short, with anterior process curved upwards and thinner apex. Tegumen long and wide, in dorsal view the basal portion of the uncus is wider, narrowing towards the tip, ending in a narrow apical process. Gnathos present, well developed, short tongue-shaped slightly up curved in lateral view (Fig. 3A), vinculum slim and proximally incurved with dorsal projection. Juxta wide in ventral view with a pointed anterior portion and a widened posterior portion as the shape of two slightly divided lobules (Fig. 3F). Aedeagus straight in lateral view, ~ 1/2 the length of the genital capsule, evenly wide, sclerotized, without cornuti (Fig. 3E).

Figure 3. 

Male genitalia of Dione (Agraulis) dodona sp. nov. A general, mesal view B distal portion of valve crista under scanning electron microscopy (pointed with open arrow in C) C right valve D termen of valve under scanning electron microscopy (pointed with closed arrow in C) E aedeagus, lateral F juxta, posterior. Scale bars: 1mm (A); 250 µm (B); 500 µm (C, E, F); 200 µm (D).

Figure 4. 

Female genitalia of Dione (Agraulis) dodona sp. nov. A external view, lateral B signum C stink club under scanning electron microscopy, specialized scales indicated by closed arrows. Asterisk indicated dorsal glands. Scale bars: 1 mm (A); 500 µm (B); 200 µm (C).

Female: Wingspan 50–52 mm, forewing length 26–27 mm. Very similar to male, but paler, with the most prominent dorsal marks and spots mainly in the forewing apex, where it presents a faint stain between the veins R₁ to M₁, absent in males and with a paler background than the rest of the wing (Fig. 1C, D). Abdomen with stink-clubs attached to a lateral fold, dorsally on posterior margin of the eighth sternum, densely covered with elongated, either single or bifid, specialized scales (Fig. 3C).

Female genitalia : Eighth segment narrow. Posterior apophysis ~ 1/2 the length of the papilla anales (Fig. 3A). Two signa slightly arched with the proximal tip near the ductus bursae, formed by four or five rows of wide spines (Fig. 3B).

Egg (Figs 5A, B, 9A, 2S)

Sub spherical, flat base slightly narrowed near apex. Yellow when recently laid (Fig. 5A), reddish brown with a whitish band subsequently, and showing larva by transparency close to hatching (Fig. 5B). Size (mean ± standard error): diameter - 0.92 ± 0.03 mm; height -1.13 ± 0.02 mm (n = 10). The chorion is adorned with 19–20 vertical and 16 or 17 horizontal carinae of smooth surfaces, which when intersect delimit cells (Fig. 9A). In the lower part of the egg, the vertical carinae are generally twice as wide as the horizontal ones and some are interrupted at one or two cells before the micropylar region. In the upper third, they have similar widths. Aeropiles scattered in the intersection of carinae, and similar in shape to those of “Agraulis vanillae”, as interpreted by da Silva et al. (2006). Micropylar region is surrounded by a rosette-like sculpture of the chorion.

Figure 5. 

Immature stages of Dione (Agraulis) dodona sp. nov. A freshly-laid egg, lateral view B egg just prior to hatching, lateral C first instar, latero-dorsal D, E fifth instar, head in detail (anterior view) and general aspect (lateral), respectively F, G, H pupa in ventral, dorsal and lateral views, respectively. Scale bars: 400 µm (A); 1 mm (C); 5 mm (E); 4 mm (H).

(Figs 5C, 6A–D, 9B, S2). Length (mean ± standard error; n = 6) = 3.56 ± 0.34 mm. Head, prothoracic dorsal shield, anal shield, pinnacles, legs and lateral plates of prolegs blackish; thorax and abdomen mostly creamy white, slightly translucent, reddish brown dorsally on A1–5 (Fig. 5C). Prothoracic dorsal shield trapezoidal with rounded angles, posterior margin with cleft at middle. Chaetotaxy as shown in Fig. 6A–D; SV group unisetose on T2–3; D2 ~ 1/2 the length of D1; and spiracles laterally on prothorax and A1–8, circular, with peritrema elevated (Fig. 9B).

Figure 6. 

Larval chaetotaxy of Dione (Agraulis) dodona sp. nov. A, B head capsule, frontal and lateral view respectively C prothoracic dorsal shield, dorsal D first instar, lateral E fifth instar, lateral. A, anterior seta; AF, adfrontal seta; C, clypeal seta; D, dorsal seta; F, frontal seta; L, lateral seta; P, postero-dorsal seta; S, stemmatal seta; SS, sub-stemmatal seta; PL, seta of proleg cylindrical section of tenth abdominal segment; PP, paraproctal seta; Sc, cephalic scolus; SD, subdorsal seta; Sl, subspiracularscolus; Sn, anal scolus; So, dorsal scolus; Sp, supraspiracularscolus; SV, subventral seta; XD, prothoracic seta. Scale bars: 200 µm (A); 100 µm (B, C); 500 µm (D); 3 mm (E).

From the second instar on, the head is black with thorax and abdomen yellow with two bands running along the subdorsal and subspiracular area. Three chromatic patterns were observed, mainly in the fifth instar, one of these patterns (brownish) is characterized by the head, legs and black scoli; thorax and abdomen yellow with brown band in the subdorsal and subspiracular area, with a thin dorsal brown line, the head has a pattern of white spots located on the frontoclypeous, and with the brown labrum, lateral plates of prolegs black (Fig. 7A); the second colored pattern (greyish) with head and black setae, thorax and abdomen with bands in the subdorsal and subspiracular area gray, the base of prolegs gray too, head with pattern of cream coloration with a larger area than the previous one and the gray labrum, with a triangular spot into the frontoclypeous, lateral plates of prolegs cream (Fig. 7B); the third pattern (reddish) is similar to second pattern but the color of the bands in abdomen, base of scoli and the head is more reddish (Fig. 7C). However, the predominant pattern observed in the field was the brownish one (Fig. 5D, E).

Figure 7. 

Variation in coloration patterns among fifth-instar of Dione (Agraulis) dodona sp. nov., shown schematically for the head capsule and fourth abdominal segment, respectively, in frontal and lateral views A brownish B grayish C reddish.

(Figs 5C, D, 6E, S2) Length (mean ± standard error; n = 5) = 25.45 ± 1.78 mm. Head blackish, covered by a large number of hair-like setae and short chalaza-like setae, which vary in length, and bears two short stout scoli dorsally (Fig. 5D). The thorax has the integument covered by conical, striated microtrichia, prominent on coxa of legs and latero-ventral face of prothorax; abdomen with cuticular sculpturing composed of irregular ribs, except the last segment ventrally on posterior face of anal proleg with conspicuous microtrichia; prothoracic dorsal shield bears a number of short chalaza-like setae and two pairs of stout spine-like setae on dorsal conical projections; elliptical spiracles with pronounced peritrema laterally on prothorax and abdominal segments A1–8, those of prothorax and A8 slightly larger than the remaining ones; prolegs with lateral plates covered by several hair-like and chalaza-like setae, crochets in uniserial and multiordinal arrangement. Thirty pairs of thoracic and abdominal scoli, which are elongated conical, integumentary outgrowths, provided with some short chalaza-like setae on the surface, one of which, typically the longest one, is placed at the apex; 11 dorsal pairs (T2–3 and A1–9); ten supra-spiracular pairs (T2–3 and A1–8), with those of the meso- and metathorax anteriorly displaced; eight lateral pairs (A1–8), and one anal pair (A10). Twelve pairs of thoracic and abdominal verrucae; three pairs on prothorax, one between dorsal shield and spiracle, which is provided with a spine-like seta, another greatly reduced pair anterior to spiracle, and another pair between spiracle and coxa; two pairs on meso and metathorax, one posterior to supraspiracular scolus and another dorsal to coxa; one pair on A1–2 and A7–8, which is ventral to lateral scolus; and one pair on A9, on the latero-ventral face of the segment.

The successive instars can be accurately distinguished by the width of the head capsule, because they do not overlap (Table 2). The corresponding exponential growth equation was adjusted for the five instars: y = 0.328eˆ0.403x; n = 62; r² = 0.980; p < 0.05. Thus, the growth pattern of the head capsule follows the Brooks-Dyar´s rule (Daly 1985). The mean growth ratio among instars was 1.495, similar to ratios previously reported for other Neotropical heliconians (Antunes et al. 2002; Kaminski et al. 2002, 2008; Tavares et al. 2002; Paim et al. 2004; da Silva et al. 2006, 2008; Barão and Moreira 2010; Vargas et al. 2014; Barão et al. 2015).

Pupa (Figs 5F–H, 8, 9D–F) General shape elongated, ground color non-uniform, consisting of a mixture of shades of gray, light brown, and ocher (Fig. 5F–H), with variation between individuals in their intensity. Length (mean ± standard error; n = 5) = 19.98 ± 0.28 mm. Head with pair of short, angled cephalic projections (Figs 8, 9D); epicranial suture absent; eyes with sculptured region near antenna, bearing few short hair-like setae, and another smooth region near front; labrum as a slight, short longitudinal stripe between the mandibles; maxilla with well-developed galeae, along midline of ventral surface, anteriorly delimited by labrum and mandibles, slightly surpassing posterior margin of A4; antennae arising laterally on head, projected ventrally to apex of maxilla, with many smooth, round tubercles on surface. Thorax with the three segments exposed. Prothorax as a small hexagonal plate in dorsal view, with anterior and posterior margins broadly excavated, with two pair of lateral tubercles. Mesothorax broadly expanded laterally along anterior half with a meso-dorsal crest that is well developed, broadly rounded, bearing a pair of round lateral tubercles; two pairs of marginal and submarginal tubercles close to base of wings; one pair of submarginal tubercles each near apex of wing; mesothoracic spiracle opening laterally at anterior margin of segment. Metathorax as a narrow plate with anterior margin broadly excavated, with pair of lateral tubercles, hindwings as straight stripes between forewings and abdominal segments. Abdomen with segments A1–A4 partially hidden by wings; with pair of lateral tubercles on A1–A7 which are little developed on A1–A2, most developed on A3, and decreasing in size posteriorly; one meso-dorsal tubercle on A5–A7; one supraspiracular tubercle on A2–A4; one pair of subspiracular tubercle on A4, pair of ventral tubercles on A5–A6; spiracles of A1 and A2 hidden and partially hidden, respectively, by forewings, and spiracles of A3–A7 elliptical; anal segment with two prominent tubercles, ventrally. Cremaster quadrate, with truncate apex, and a large number of short, curved hooks.

Figure 8. 

Pupa of Dione (Agraulis) dodona sp. nov. under A ventral B dorsal, and C lateral view, respectively. A, abdominal segment; T, thoracic segment. Scale bar: 4 mm.

Figure 9. 

Immature stages of Dione (Agraulis) dodona sp. nov. under scanning electron microscopy A egg, lateral view B first instar, upper portion of fifth abdominal segment, lateral C fifth instar, prothoracic plate, postero-dorsal (protuberance pointed by close arrow) D, E, F pupal head (dorsal view), mesothoracic meso-dorsal crest (lateral), and latero-dorsal tubercle of third abdominal segment (lateral), respectively. Scale bars: 500 µm (A, F); 100 µm (B); 250 µm (C); 1.5 mm (E, D).

The specific epithet is based on the locality of Dodona (Greece); it was a city-sanctuary in ancient Greece, where there was an oracle in which Dione was venerated as the (temporary) wife of Zeus, until she was replaced by Hera. Thus, the new species is named “dodona” to continue the classical Greek tradition.

Adults of Dione (Agraulis) dodona sp. nov. are known from distinct populations, located in central and southern Peru, and northern Chile, on the western slopes of the Andes. In Peru, it has been found in the Departments of Lima (1,400–2,400 m elevation), Arequipa (between 800 and 2,600 m elevation), Moquegua (1,800–2,100 m elevation), and Tacna (1,800 m elevation). In Chile, two specimens were collected in 1951 and 1968 from two localities in the Tarapacá Region between 2,300 and 3,000 m elevation, and recently other two specimens were collected from Arica Region (1,580 m elevation).

Malesherbia tenuifolia D. Don (Passifloraceae) is the only host plant known for the immature stages of Dione (Agraulis) dodona sp. nov. This species was described originally from northern Chile. It is distributed between 19–21 °S in Chile (Bull-Hereñu 2020), and was reported recently from southern Peru (Weigend et al. 2015; Beltrán et al. 2018), restricted to the western slopes of the Andes above 1,500 m. M. tenuifolia is a shrub with reddish or yellowish tubular flowers that grows to ca. 1 m in height (Fig. 10), associated with the dry beds and immediate surroundings of seasonal rivers in Chile (Bull-Hereñu 2020), and steep scree slopes along the road cuts in the type locality of Dione (Agraulis) dodona sp. nov. (Fig. 10A).

Figure 10. 

Life history of Dione (Agraulis) dodona sp. nov. A general view of type locality, Pacaychacra valley, Arequipa, Peru (blue close arrow points to larval hostplant) B host plant, Malesherbia tenuifolia Don C female laying eggs (close arrow point) on flowers of host plant D eggs (pointed by setae) on flower E detail of flower showing damage (entrance) by first instar larva (open arrow). Scale bar: 5 mm.

Adults of Dione (Agraulis) dodona sp. nov. were only observed on sunny days in the type locality, beginning to fly around 08:00 a.m., quickly disappearing when weather conditions became cloudy. They usually fly close to the ground, up to 1–2 m high. Females were seen alighting on the host plant. Territorial behavior, courtship and mating behavior were not observed. Adults were seen feeding on flowers of other plants growing around Malesherbia hosts. The species is multivoltine in the population of the type locality, flying all year round. From three to seven individuals were usually observed in a typical sunny day (4–5 h of observation), most of which were males. This species is sympatric with Dione (Agraulis) forbesi, but they do not fly in the same habitat; forbesi was observed in areas with higher humidity and vegetation, compared to dodona, which was found only in xerophytic areas associated with the host plant (Fig. 10A, B). Females were observed laying eggs particularly on flowers (Fig. 10D). Oviposition occurred with the female sitting on top of the flower and curling the abdomen around the flower edge so that eggs were deposited underneath (Fig. 10C). During visits made at different months of the year in the type locality, freshly laid eggs were always collected, both on flowers and leaves. Between one and eight eggs were found in the same plant, in different flowers or leaves, but sometimes several eggs were obtained from the same flower (Fig. 10D). The eggs are laid isolated from each other. Newly hatched larvae first consumed the chorion, and afterwards began to feed inside of flower, leaving a hole through which they enter the flower (Fig. 10E). Subsequent instars feed externally on leaves. Larvae were consistently solitary in all instars, regarding all activities, such as feeding or resting. Pupae were found predominantly off the host plant, clinging to rocks near the host plant, and sometimes on branches of the host plant close to the ground.

Dione (Agraulis) dodona sp. nov. was recovered as an independent lineage within the Agraulis clade of the COI-tree (Fig. 11), diverging in ca. 5% to the group formed by its other species, and 9.5% to the Dione clade (Table 3).

Figure 11. 

Phylogenetic status of Dione (Agraulis) dodona sp. nov. based on 650 bp-sequences of the Cytochrome oxidase subunit I gene. The consensus tree was inferred with the Maximum Likelihood method and General Time Reversible model, with 500 bootstrap replicates. The branch support (bootstrap) is shown next to the nodes.