Phylogenetic analysis of the sharpshooter genus Subrasaca Young, 1977 (Hemiptera, Cicadellidae, Cicadellini).

The South American sharpshooter genus Subrasaca comprises 14 species. Some species of this genus are quite common in the Brazilian Atlantic Rainforest. In this paper, a phylogenetic analysis of Subrasaca, based on a matrix of 20 terminal taxa and 72 morphological characters of the head, thorax, and male and female genitalia, is presented. The analysis yielded six equally most parsimonious trees (197 steps, CI = 0.6091, RI = 0.5722, and RC = 0.3486). The results suggest that Subrasaca is a monophyletic taxon, although the genus branch is not robust. The clade showing the highest bootstrap and Bremer scores is formed by species with longitudinal dark brown to black stripes on the forewings (Subrasacabimaculata, Subrasacaconstricta, Subrasacacurvovittata, and Subrasacaflavolineata), followed by Subrasacaatronasa + Subrasacaaustera.

Introduction

The infraorder comprises three superfamilies, (cicadas), (spittlebugs or froghoppers), and (leafhoppers and treehoppers). According to Hamilton (1999), the monophyly of the is well-supported by morphological synapomorphies, including the presence of a complex filter chamber. Cryan (2005), based on molecular data (18S rDNA, 28S rDNA, and histone 3), also supports the monophyly of the and suggests the following relationships for the superfamilies: ( (, )). Based both on morphological and molecular data, the monophyly of the is also well-supported (Evans 1963, Dietrich and Deitz 1993, Hamilton 1999, Cryan 2005). Synapomorphies of the include the enlarged, transverse metathoracic coxae and a pair of rod-shaped lateral apodemes associated with the scutellar suture (Dietrich and Deitz 1993).

The family (leafhoppers), with over 21,000 described species placed in more than 120 family-group taxa (Oman et al. 1990, Hamilton 1999), includes many species of economic importance because they are vectors of pathogens of cultivated plants (Nielson 1985). According to the morphological phylogeny of Hamilton (1983) and the molecular phylogeny (28S rDNA) of Dietrich et al. (2001), is a paraphyletic group because treehoppers ( and ) are derived from leafhoppers. Taxonomically, cicadellids can be distinguished from other membracoids by the mesanepisternum without a hooklike process, separated from the katepisternum by a suture, and hind tibia with setae of longitudinal rows usually large and conspicuous (Dietrich 2005). With over 2,000 known species and a cosmopolitan distribution, (sharpshooters) is the third largest subfamily of the (Mejdalani 1998, Takiya 2007, McKamey 2007). According to Young (1968, 1977, 1986), this subfamily is divided into two tribes, a cosmopolitan and a New World . Sharpshooters feed on the low-nutrient xylem sap of vascular plants. Some species of this group are important vectors of xylem-borne phytopathogenic bacteria (Redak et al. 2004).

The genus Young, 1977 belongs to the . has records from Brazil and Argentina, as well as dubious records of from Colombia (Young 1977, McKamey 2007, Silva et al. 2013b). Species records are mostly from the Atlantic Rainforest. comprises currently 14 species (Silva et al. 2013a,b): Young, 1977, Young, 1977, Silva, Cavichioli & Mejdalani, 2013a, Silva, Cavichioli & Mejdalani, 2013a, (Stål, 1862), Silva, Cavichioli & Mejdalani, 2013b, (Signoret, 1855), (Stål, 1862), (Signoret, 1854) (type species), (Melichar, 1951), (Signoret, 1855), Silva, Cavichioli & Mejdalani, 2013b, (Signoret, 1854), and Silva, Cavichioli & Mejdalani, 2013b.

Taxonomically, PageBreak differs from other genera of the by the following combination of male genital characteristics (Silva et al. 2013b): (1) aedeagus usually short and dorsally expanded; (2) styles (parameres) with distinct preapical lobe; (3) paraphyses with two or four rami (except in , with only one ramus); and (4) subgenital plates connected to each other at base by a triangular membranous area, not extending posteriorly as far as pygofer apex. species are generally quite colorful and range in length from 4.8 to 7.7 mm. Young (1977: 445), based on overall similarity, included in his group of genera, which also includes Melichar, 1926, Melichar, 1926, Young, 1977, Young, 1977, Young, 1977, Young, 1977, Melichar, 1926, Young, 1977, and Young, 1977.

Here we use morphological data of the head, thorax, male and female genitalia to investigate the phylogenetic relationships among the species of . Among our outgroups, we included four genera of the group (, , , and ).

Material and methods

Specimens for the study

Specimens of 12 of the 14 described species of were studied ( and were not obtained and thus coded based on Young 1977 and Wilson et al. 2009). The matrix includes 20 terminal taxa (14 species and six outgroups). The outgroups are four representatives of the generic group [ Mejdalani & Nessimian, 1991, sp., (Taschenberg, 1884), (Stål, 1862)], (Walker, 1851), and a member of the , Melichar, 1926, which was employed for rooting the trees.

The studied specimens belong to the following institutions: Departamento de Entomologia, Museu Nacional, Universidade Federal do Rio de Janeiro (MNRJ, Rio de Janeiro); Coleção Entomológica Prof. José Alfredo P. Dutra, Departamento de Zoologia, Instituto de Biologia, Universidade Federal do Rio de Janeiro (DZRJ, Rio de Janeiro); and Coleção de Entomologia Pe. Jesus S. Moure, Departamento de Zoologia, Setor de Ciências Biológicas, Universidade Federal do Paraná (DZUP, Curitiba). The number of specimens examined of each terminal taxon, their geographical distribution, and collections are listed in Table 1.

Table 1.

Taxa included in the phylogenetic analysis of (in bold) and outgroups. The number of females and males examined, their distribution (Brazilian states), and collections are provided for each taxon.

Taxon	Females	Males	Distribution	Collection
Cyclogonia caeliguttata Mejdalani & Nessimian, 1991	2	2	RJ	MNRJ
Juliaca sp.	2	2	RJ	MNRJ
Geitogonalia quatuordecimmaculata (Taschenberg, 1884)	2	2	RJ	MNRJ
Scopogonalia subolivacea (Stål, 1862)	2	2	RJ, MG	MNRJ
Versigonalia ruficauda (Walker, 1851)	2	2	RJ	MNRJ
Tretogonia cribrata Melichar, 1926*	2	2	RJ	MNRJ
Subrasaca atronasa Young, 1977**	–	–	–	–
Subrasaca austera Young, 1977	2	1	SC	DZUP
Subrasaca bimaculata Silva et al., 2013	19	26	MG, SP, PR	DZRJ, DZUP, MNRJ
Subrasaca constricta Silva et al., 2013	2	3	BA	DZUP, MNRJ
Subrasaca curvovittata (Stål, 1862)	11	6	RJ	DZRJ, DZUP, MNRJ
Subrasaca diminuta Silva et al., 2013	8	6	SP, PR	DZUP, MNRJ
Subrasaca flavolineata (Signoret, 1855)	9	15	RJ	DZRJ, DZUP, MNRJ
Subrasaca flavoornata (Stål, 1862)	6	2	RJ	MNRJ
Subrasaca ignicolor (Signoret, 1854)	22	14	RJ, SP	MNRJ
Subrasaca monacha (Melichar, 1951)**	–	–	–	–
Subrasaca nigriventris (Signoret, 1855)	10	10	RJ	MNRJ
Subrasaca rachelae Silva et al., 2013	19	12	ES	DZRJ, DZUP, MNRJ
Subrasaca rhienetta (Signoret, 1854)	3	3	RJ, SP	MNRJ
Subrasaca rubra Silva et al., 2013	7	10	MG, RJ, SP	DZRJ, DZUP, MNRJ

Brazilian states: BA – Bahia; ES – Espírito Santo; MG – Minas Gerais; PR – Paraná; RJ – Rio de Janeiro; SC – Santa Catarina; SP – São Paulo. DZRJ – Departamento de Zoologia, Universidade Federal do Rio de Janeiro; DZUP – Departamento de Zoologia, Universidade Federal do Paraná; MNRJ – Museu Nacional, Universidade Federal do Rio de Janeiro; * root of the phylogenetic analysis; ** coded based on Young (1977) and Wilson et al. (2009).

Techniques for preparation of specimens and terminology

The techniques for preparation of male and female genital structures follow Oman (1949) and Mejdalani (1998), respectively. The dissected parts are stored in small vials with glycerin, as suggested by Young and Beirne (1958). The first and second pair of valvulae of the ovipositor were mounted on temporary slides with glycerin. The descriptive terminology adopted herein follows mainly Young (1977), except for the facial areas of the head (Hamilton 1981, Mejdalani 1993, 1998) and the female genitalia (Nielson 1965, Hill 1970).

Cladistic analysis

Morphological characters of the head, thorax, male and female genitalia were included in the unpolarized matrix (Nixon and Carpenter 1993), which was assembled using the PageBreakNexus Data Editor (Page 2001). Hypotheses of primary homology were proposed based on the topological identity of the structures (Pinna 1991). All characters were initially scored equal weights. Character states were scored as underscores (_) when inapplicable or as question marks (?) when unavailable. The heuristic search algorithm, as implemented in PAUP* 4.0 (Swofford 2002), was employed for searching the most parsimonious trees. The successive weighting procedure (Carpenter 1988, 1994) was based on the maximum rescaled consistency index (rc) of the characters (Farris 1969, 1989). The strict consensus method was employed for all original most parsimonious trees. Clade support was estimated by computing 10.000 bootstrap replicates (Felsenstein 1985) with heuristic search in PAUP* 4.0 and by decay indices (Bremer 1988, 1994) in TreeRot 3.0 (Sorenson and Franzosa 2007). Autapomorphic characters were included in the matrix, as suggested by Yeates (1992), but we provide consistency index (CI) values considering all characters as well as only the informative ones.

Results and discussion

The data matrix (Table 2) consists of 72 morphological characters, 35 of the external morphology, 25 of the male genitalia, and 12 of the female genitalia. Among these characters, 51 are binary and 21 are multistate, being 52 informative for the parsimony analysis. The characters, their states, and ci greater than 0.5 are listed below. Although many of the characters are based on color patterns, these are consistent intraspecifically in . Figures 1 (external morphology and male genitalia) and 2 (female genitalia) provide some examples of characters employed in the phylogenetic analysis.

Table 2.

Data matrix for the phylogenetic analysis of (in bold) and outgroup taxa. (_) codes for inapplicable states, (?) for unavailable data, (A) for state 10, (B) for 11, and (C) for 12. Outgroup genera are , , , , , and (root).

Taxa	Characters
	1 2 3 4 5 6 7
	1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2
Cyclogonia caeliguttata	0 2 1 0 _ 0 0 1 2 0 1 0 0 0 _ 0 _ 0 1 2 0 5 0 3 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 0 0 1 0 1 1 2 1 0 _ 1 0 _ _ 0 0 1 1 _ 0 0 _ 0 1 0 0 _ _ _ 0 4 1 1 0 0
Juliaca sp.	1 1 1 0 _ 0 0 0 _ _ _ 1 1 0 _ 0 _ 1 0 _ _ 6 0 1 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 0 1 0 _ 1 1 1 1 0 _ 1 0 _ _ 0 0 1 1 _ 0 0 _ 0 3 0 0 _ _ _ 0 2 1 1 0 1
Geitogonalia quatuordecimmaculata	0 1 4 1 2 0 0 1 0 0 0 0 0 0 _ 0 _ 0 1 0 1 0 0 2 0 _ _ 0 _ _ _ 0 _ 1 1 0 0 0 0 1 1 1 0 2 1 0 _ 1 1 1 0 0 0 1 1 _ 0 0 _ 0 2 0 0 _ _ _ 0 3 0 1 0 0
Scopogonalia subolivacea	1 2 C 0 _ 0 0 0 _ _ _ 0 0 0 _ 0 _ 0 0 _ _ 9 0 1 0 _ _ 0 _ _ _ 0 _ 1 0 0 1 1 0 0 _ 0 1 _ _ 0 _ 1 1 1 2 0 0 1 1 _ 0 0 _ 0 0 0 1 0 1 2 0 4 0 1 0 0
Versigonalia ruficauda	0 2 B 0 _ 1 0 0 _ _ _ 0 1 0 _ 0 _ 0 0 _ _ 8 0 4 0 _ _ 1 0 2 0 0 _ 1 2 0 1 0 0 0 _ 1 1 1 1 0 _ 1 0 _ _ 0 1 0 _ _ _ _ _ _ 0 0 0 _ _ _ 0 4 0 1 1 2
Tretogonia cribrata (root)	0 2 2 0 _ 0 0 0 _ _ _ 0 0 0 _ 0 _ 0 0 _ _ 5 0 3 0 _ _ 0 _ _ _ 0 _ 0 1 0 1 0 1 1 0 3 1 1 1 0 _ 1 0 _ _ 0 1 0 _ _ _ _ _ _ 0 0 1 0 0 5 1 0 0 0 0 0
Subrasaca ignicolor	0 0 6 0 _ 0 0 1 0 1 0 0 0 0 _ 0 _ 0 1 0 0 0 0 2 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 1 2 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 0 0 _ 0 0 0 1 1 1 2 0 4 0 0 0 0
Subrasaca rhienetta	1 2 3 1 0 0 1 0 _ _ _ 1 1 0 _ 0 _ 1 0 _ _ 0 0 0 0 _ _ 0 _ _ _ 1 0 1 0 0 0 1 0 1 0 2 1 1 1 1 1 0 0 _ _ 0 0 1 1 _ 0 0 _ 0 0 1 1 1 0 4 0 4 0 1 0 1
Subrasaca nigriventris	0 0 1 0 _ 1 0 1 0 1 1 0 0 0 _ 0 _ 0 1 0 2 0 0 0 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 0 0 1 1 1 0 _ 1 1 1 2 0 0 1 1 _ 0 0 _ 0 0 1 1 1 0 0 0 4 0 0 0 0
Subrasaca flavolineata	1 1 8 1 1 0 1 0 _ _ _ 1 1 0 _ 1 1 0 0 _ _ 2 0 1 1 0 0 1 0 0 0 0 _ 1 0 0 0 1 0 1 1 1 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 0 1 1 0 0 1 1 1 1 1 0 4 1 0 0 0
	1
Subrasaca monacha	0 0 2 0 _ 0 0 1 1 1 0 0 0 0 _ 0 _ 0 0 _ _ 5 0 3 0 _ _ 0 _ _ _ 0 _ 1 ? 0 0 ? 0 1 0 2 1 1 1 0 _ 1 1 0 3 0 0 1 0 _ 0 0 _ 0 ? ? ? ? ? ? ? ? ? ? ? ?
Subrasaca flavoornata	0 0 9 0 _ 0 0 1 0 1 0 0 0 0 _ 0 _ 0 1 0 3 0 0 0 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 0 1 1 0 1 0 _ 1 1 1 1 0 0 1 1 _ 0 0 1 1 1 0 1 1 1 4 1 4 0 1 0 0
	3 1
Subrasaca curvovittata	1 1 7 1 0 0 1 0 _ _ _ 1 1 0 _ 1 1 0 0 _ _ 1 0 0 1 0 0 1 1 0 0 0 _ 1 0 0 0 1 0 1 0 1 1 1 1 1 0 0 0 _ _ 0 0 1 2 0 0 0 _ 0 0 1 1 1 1 4 1 6 1 0 0 0
	1
Subrasaca atronasa	1 0 ? 0 _ 0 0 1 2 1 0 0 0 1 0 0 _ 0 0 _ _ 5 1 1 0 _ _ 0 _ _ _ 1 1 1 0 0 0 1 0 1 0 2 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 0 0 _ 0 ? ? ? ? ? ? ? ? ? ? ? ?
Subrasaca austere	1 0 A 1 3 0 0 1 2 1 0 0 0 1 1 0 _ 0 0 _ _ 5 1 3 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 0 2 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 0 0 _ 0 0 0 1 1 0 0 0 4 0 1 0 1
	1
Subrasaca constricta	1 0 1 0 _ 0 1 0 _ _ _ 1 1 0 _ 1 0 0 0 _ _ 7 0 1 1 2 1 1 0 0 1 0 _ 1 0 1 0 1 0 1 0 0 1 1 0 1 1 0 0 _ _ 0 0 1 1 _ 0 1 0 0 0 0 1 1 1 0 0 5 0 0 0 0
Subrasaca bimaculata	1 1 6 1 0 0 1 0 _ _ _ 1 1 0 _ 1 1 0 0 _ _ 4 0 1 1 1 0 1 0 0 0 0 _ 1 0 0 0 1 0 1 0 0 1 1 1 1 0 0 0 _ _ 0 0 1 2 1 0 0 _ 0 1 0 0 _ _ _ 0 1 1 0 0 0
	1 1 2
Subrasaca rachelae	0 0 1 0 _ 1 0 1 0 1 1 0 0 0 _ 0 _ 0 1 0 2 3 0 3 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 0 0 1 1 0 1 0 0 0 _ _ 1 0 1 1 _ 0 0 _ 1 1 1 1 1 0 2 0 4 0 1 0 0
Subrasaca diminuta	0 0 5 0 _ 0 0 1 0 1 0 0 0 0 _ 0 _ 0 1 0 0 0 0 0 0 _ _ 0 _ _ _ 0 _ 1 0 0 0 1 0 1 0 2 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 1 0 _ 0 0 0 1 1 0 2 0 4 0 0 0 0
Subrasaca rubra	0 0 0 0 _ 0 0 0 _ _ _ 0 0 0 _ 0 _ 0 1 0 3 0 0 2 0 _ _ 0 _ _ _ 0 _ 1 1 0 0 1 0 1 0 0 1 1 1 1 0 0 0 _ _ 0 0 1 1 _ 0 0 _ 0 0 0 1 1 1 3 1 4 0 1 1 1

Figure 1.

Examples of characters for the phylogenetic analysis of (external morphology and male genitalia). a body of (length 5.3 mm): rounded anterior margin of crown (character 1, state 0), maculae on lateroapical portions of crown (c6, s1), pair of moderately oblique maculae on pronotum (c21, s2) b (length 5.4 mm): mesonotum with T-shaped macula (c22, s2), longitudinal stripes on forewings (c25, s1) c (length 5.7 mm): pronounced anterior margin of crown (c1, s1) d pygofer lobe of , dorsal view: dorsoapical process (c36, s1; arrowed) e subgenital plates of : membranous basal area (c38, s1; arrowed) f : styles with preapical lobe (c40, s1) and apex transversely truncate (c42, s0), stalk of connective clearly differentiated, not extending beyond apex of styles (c44, s1) g aedeagus of : dorsal lobe (c46, s1) with constriction (c47, s1; arrowed) h aedeagus of : shaft longer than high (c48, s1), pair of spiniform apical processes (c51, s2) i aedeagus of : pair of preapical processes (c52, s1; arrowed) j : paraphyses with two rami (c55, s1) k : paraphyses with four rami (c55, s2), inner rami small and narrow (c56, s0) l : inner rami of paraphyses broader and larger than outer rami (c56, s1).

Figure 2.

Examples of characters for the phylogenetic analysis of (female genitalia). a sternite VII of : convex posterior margin at median portion (character 61, state 1) b sternite VII of : concave posterior margin at median portion (c61, s0) c : smooth sclerites of “inner” sternite VIII (c65, s0) with linear aspect (c66, s2; arrowed) d : sclerites of “inner” sternite VIII with oblique aspect (c66, s4; arrowed) e : punctuated sclerites of “inner” sternite VIII (c65, s1; arrowed) f sp.: valvifer I subrectangular (c68, s2) g : valvifer I ellipsoid (c68, s4) h : valvifer I gutiform (c68, s6) i : valvulae I with expanded base (c69, s0) j : valvula II with obtuse apex (c70, s0) and convex dorsal margin (c72, s0) k : valvula II with acute apex (c70, s1), linear and indistinct teeth (c71, s1), and rectilinear dorsal margin (c72, s1).

Morphological characters of the phylogenetic analysis

External morphology and coloration

Main aspects and discussion of the phylogenetic analysis

The analysis with equal weights resulted in six most parsimonious trees with length = 197, consistency index (CI) = 0.6091 (excluding uninformative characters = 0.5389), retention index (RI) = 0.5722, and rescaled consistency index (RC) = 0.3486. The trees differ from one another (1) in the position of (outgroup), (2) positions of and , which appear as sister groups or not, and (3) positions of , , , and . These four species formed a clade with + in two trees. A strict consensus of the six trees is given in Fig. 3a.

Figure 3.

a Strict consensus of the six equally most parsimonious trees of the phylogenetic analysis of and outgroup taxa b Most parsimonious tree obtained with the successive weighting procedure; length = 80, consistency index = 0.8249 (excluding uninformative characters = 0.7199), retention index = 0.7641, rescaled consistency index = 0.6303. Outgroup genera are , , , , , and (root).

The successive weighting analysis yielded one tree, which is also one of the six original trees, with length = 80, CI = 0.8249 (excluding uninformative characters = 0.7199), RI = 0.7641, and RC = 0.6303 (Figs 3b, 4). Thirty-two characters had maximum weight (= 1.0) and 40 had lower weights. Twenty characters were parsimony-uninformative. Figure 4 gives bootstrap estimates (when > 50%) and Bremer support indices for the clades recovered under equal weights. Apomorphies of this tree are given in Table 3.

Figure 4.

One of the most parsimonious trees of the phylogenetic analysis of and outgroup taxa; this is also the single tree obtained with the successive weighting procedure. Length = 197, consistency index = 0.6091 (excluding uninformative characters = 0.5389), retention index = 0.5722, rescaled consistency index = 0.3486. Species of in bold. Apomorphies are given in Table 3. Most sharpshooter images from Wilson et al. (2009).

Table 3.

Apomorphy list for clades of Fig. 4 of the phylogenetic analysis of and outgroup taxa. Non-homoplastic characters are in bold.

Node or terminal taxon	Apomorphies
37	37(0), 53(0), 54(1)
21	1(1), 22(6), 24(1), 65(1)
36	8(1), 40(1), 64(1)
22	5(2), 19(1), 44(2), 51(0), 61(1)
35 (Subrasaca)	2(0), 10(1), 38(1), 42(2), 63(1)
34	46(1), 48(0)
33	1(1), 3(A), 5(3), 9(2), 14(1), 23(1), 33(1), 66(0), 72(1)
32	19(1), 22(0), 24(0)
23	3(5), 70(0)
31	21(2), 42(0)
30	6(1), 11(1), 62(1)
29	21(3), 51(1), 65(1), 66(4), 67(1)
28	8(0), 72(1)
27	1(1), 4(1), 7(1), 12(1), 13(1), 19(0), 47(1), 67(0)
26	16(1), 22(1), 24(1), 25(1), 28(1), 70(0), 72(0)
25	2(1), 3(8), 17(1), 47(0), 55(2), 59(1), 69(1)
24	42(1), 62(1)
Versigonalia ruficauda	3(B), 6(1), 13(1), 22(8), 24(4), 28(1), 30(2), 35(2), 71(1), 72(2)
Juliaca sp.	2(1), 12(1), 13(1), 18(1), 39(1), 61(3), 68(2), 69(1), 72(1)
Scopogonalia subolivacea	3(C), 22(9), 37(1), 38(1), 42(0), 49(1), 63(1)
Geitogonalia quatuordecimmaculata	2(1), 3(4), 4(1), 21(1), 22(0), 24(2), 35(1), 41(1), 43(0), 49(1), 61(2), 68(3)
Cyclogonia caeliguttata	9(2), 11(1), 20(2), 69(1)
Subrasaca monacha	3(2), 9(1), 49(1), 50(0), 51(3), 55(0)
Subrasaca atronasa	24(1), 32(1)
Subrasaca austera	4(1), 15(1)
Subrasaca ignicolor	3(6), 24(2), 41(1), 65(1)
Subrasaca diminuta	57(1)
Subrasaca nigriventris	46(0), 48(1), 49(1), 66(0), 70(0)
Subrasaca rachelae	22(3), 24(3), 45(0), 52(1), 60(1), 61(1)
Subrasaca flavoornata	3(9), 42(1), 44(0), 46(0), 48(1), 49(1), 60(1), 61(1)
Subrasaca rubra	3(0), 24(2), 35(1), 66(3), 71(1)
Subrasaca rhienetta	2(2), 3(3), 18(1), 32(1), 42(2), 62(1), 65(0)
Subrasaca constricta	4(0), 22(7), 26(2), 27(1), 31(1), 36(1), 45(0), 58(1), 66(0), 68(5)
Subrasaca bimaculata	22(4), 26(1), 56(1,2), 61(1), 63(0), 68(1)
Subrasaca flavolineata	5(1), 22(2), 41(1), 55(1), 58(1), 66(1)
Subrasaca curvovittata	3(7), 29(1), 67(1), 68(6)

The monophyly of was recovered in all most parsimonious trees (Fig. 3a). This clade, however, is not robust (bootstrap < 50%, Bremer = 0) (Fig. 4). Phylogenetically, can be tentatively defined by the following synapomorphic traits of its groundplan: (1) ocelli located slightly anterad of the imaginary line between the anterior angles of eyes (character 2, state 0; Fig. 1c), (2) complete transverse band on middle portion of crown (character 10, state 1), (3) triangular membranous area uniting subgenital plates basally (character 38, state 1; Fig. 1e), (4) obtuse shape of apex of styles in dorsal view (character 42, state 2), and (5) sclerites of female “inner” sternite VIII present (character 63, state 1; Fig. 2c–e).

Other phylogenetic (e.g., Felix and Mejdalani 2011) or purely taxonomic (e.g., Mejdalani et al. 2014) studies on the highlighted the need for more precise definitions of various genera of this tribe. In the introduction of his impressive monograph on the New World , Young (1977: 10) expressed his perception of this problem as follows: “The are an intricate group. Their morphology suggests rapid radiation and often shows small discontinuities compared with those found in many of the .” Small discontinuities are precisely what we have observed between and the genera here employed as outgroups. In any case, the cladistic analysis allowed us to propose a more objective definition of the genus.

Two clades appeared in all six most parsimonious trees and were fairly robust in the analysis (Fig. 4). The clade formed by the species with longitudinal dark brown PageBreakto black stripes on the forewings (PageBreakPageBreak, , , and ) had the highest percentage of bootstrap (= 83%) and was supported by seven apomorphic conditions (Table 3, node 26), including the conspicuous set of dark brown to black longitudinal stripes on the forewings (character 25, state 1; Fig. 1b, c). The Bremer support of this clade was 3. It shows the following internal relationships in all trees (Fig. 3a): ( ( (, ))). These four species were described in detail by Silva et al. (2013a). The group is distributed in the Atlantic Forest from northeastern (state of Bahia) to southern Brazil (state of Paraná). The second clade, formed by + , is supported by nine apomorphic conditions (Table 3, node 33), including a whitish-yellow transverse band on middle portion of pronotum (character 14, state 1; Fig. 4, node 33). This clade also had relatively high bootstrap (= 82%) and Bremer (= 2) scores. These two species, which are known only from the state of Santa Catarina (Zanol and de Menezes 1982) in southern Brazil (Atlantic Forest), were described by Young (1977), who considered them “very close” to each other (Young 1977: 479).

Although with low support scores, the clades formed by + and + were recovered in all most parsimonious trees (Fig. 4, nodes 23 and 30, respectively). Unlike the species with longitudinal dark brown to black stripes on the forewings (node 26), those with contrasting orange marks ( + , + , and ) did not form a monophyletic group in any of the six most parsimonious trees.