Combining target enrichment and Sanger sequencing data to clarify the systematics of the diverse Neotropical butterfly subtribe Euptychiina (Nymphalidae, Satyrinae) Author Espeland, Marianne Author Nakahara, Shinichi Author Zacca, Thamara Author Barbosa, Eduardo P. Author Huertas, Blanca Author Marín, Mario A. Author Lamas, Gerardo Author Benmesbah, Mohamed Author Brévignon, Christian Author Casagrande, Mirna M. Author Fåhraeus, Christer Author Grishin, Nick Author Kawahara, Akito Y. Author Mielke, Olaf H. H. Author Miller, Jacqueline Y. Author Nakamura, Ichiro Author Navas, Vanessa Author Patrusky, Brooke Author Pyrcz, Tomasz W. Author Richards, Lindsay Author Tan, Denise Author Tyler, Stephanie Author Viloria, Angel Author Warren, Andrew D. Author Xiao, Lei Author Freitas, André V. L. Author Willmott, Keith R. text Zoological Research 2023 2023-02-15 2023 1 73 journal article 58039 10.1111/syen.12590 bfb878f3-8a74-46d3-a104-36485c32aaba 7909395 Euptychia Euptychia is strongly supported as a sister to the remainder of the subtribe ( Figures 2, S 2 and S 3 ) as also found in Espeland et al. (2019a) . The systematic placement of Euptychia has been unstable in previous phylogenetic studies ( Murray & Prowell, 2005 ; Peña et al., 2010 , 2011 ). Currently, 37 valid species are recognized ( Fratello et al., 2015 ; Freitas, Wahlberg, et al., 2012; Nakahara et al., 2014 , 2017 ; Nakahara, Hall, et al., 2015; Nakahara, Janzen, et al., 2015; Nakahara, Llorente-Bousquets, et al., 2015; Nakahara, Vega, & Willmott, 2016; Neild et al., 2014 , 2015 ). Only a subset of species was included here, and several taxonomic changes, in addition to the description of over a dozen species, will be made in a forthcoming revision of the genus, and thus the genus is estimated to include more than 50 species (Nakahara et al., in preparation). Members of Euptychia are relatively small for the subtribe, with the forewing length often less than 20 mm . Except for a few species, they have sexually monomorphic wing patterns often marked prominently with ventral bands and submarginal eyespots and many species exhibit translucence. Possible synapomorphies and/or distinctive characters for Euptychia include the following: (a) Third segment of labial palpi shorter than one-fourth of second segment in length; (b) absence of tibial spurs in midleg and hindleg; (c) absence of lateral spines on tibia; (d) presence of the forewing recurrent vein in the discal cell; (e) absence of basal swelling of the forewing cubital vein; (f) humeral vein relatively reduced in comparison with other Euptychiina ; (g) developed male eighth tergite and sternite; (h) presence of the sclerotized region of the eighth abdominal segment in the female, located at the very basal side of the eighth abdominal segment; (i) absence of the lateral sclerotization of the 8th abdominal segment of the female; (j) origin of the ductus seminalis at the posterior end of the ductus bursae In fact, many of these characters are unique to Euptychia (Nakahara, unpublished data), highlighting the distinctiveness of this lineage within the subtribe. Members of Euptychia range from north-western Mexico to south-eastern Brazil, and somewhat unusually for the subtribe, the genus appears to be more diverse in lower montane regions and adjacent lowlands than in lowland forest far from mountains. Adult males of some species exhibit perching behaviour, often with several individuals in close proximity, mostly in light gaps or sunflecks and often on hilltops. Although most known host plants for Euptychiina are either bamboo or other grasses in the family Poaceae , larvae of Euptychia notably feed instead on some species of Selaginella (Lycopsida) , as well as a single record utilizing Neckeropsis ( Neckeraceae ) ( Brévignon, 2008 ; Freitas, Mota, Barbosa, & Carreira, 2019; Hamm & Fordyce, 2016 ; Singer et al., 1971 ; Singer & Mallet, 1986 ). FIGURE 3 Phylogeny of the Cyllopsis clade based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 4 Phylogeny of the Megisto clade based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 5 Phylogeny of the Hermeuptychia clade and nearby genera based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 6 Phylogeny of the Amphidecta clade based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 7 Phylogeny of the ’ Taygetis clade ’ based on the FULL dataset. Interpretation of the support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 8 Phylogeny of the ’ Archeuptychia clade ’ + Chloreuptychia based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfy images equal 1 cm. FIGURE 9 Phylogeny of the ’ Pareuptychia clade ’ based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 10 Phylogeny of the ’ Splendeuptychia clade ’ based on the FULL dataset. Interpretation of support values can be found in the legend. The scale bars under the butterfly images equal 1 cm. FIGURE 11 Lazulina hewitsonii comb.n. (a) male wing venation; (b) male forewing androconial scales posterior of cubital vein; (c and d) male dorsal (c) and ventral (d); (e and f) female dorsal (e) and ventral (f); (g) male abdomen lateral view. Scale bars 1 cm unless otherwise noted. FIGURE 12 Lazulina hewitsonii comb.n. A-D, male genitalia (dissection KW-15-21), lateral (a), dorsal (b), aedeagus lateral (c) and aedeagus dorsal (d); (e and f) female genitalia (dissection SN-17-207), ventral view exterior tip abdomen (e), dorsal view interior abdomen (f). Scale bars 1 mm. FIGURE 13 Lazulina gen.n. Species diversity mapped on a 2 degree grid. Colours ranging from dark green to red represent increasing diversity. FIGURE 14 Saurona triangula comb.n. (a) male wing venation — stippled area on FW shows dorsal androconial scales around vein M 2 and ventral androconial scales around vein 2A, androconial hair-like scales at anterior edge discal cell are also indicated; (b and c) male dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 15 Saurona triangula comb.n. A-F, male genitalia (dissection KW-21-31), lateral tip abdomen (a), lateral (b) with posterior view juxta, dorsal (c), ventral (d), aedeagus lateral (e) and dorsal (f); (g – j), female genitalia (dissection USNM loan 2090579), lateral view exterior tip abdomen (g), ventral view exterior tip abdomen (h), dorsal view interior abdomen (i), corpus bursae perpendicular to signa (j).Scale bars 1 mm. FIGURE 16 Saurona gen.n. Species diversity mapped on a 2 degree grid. Colours ranging from dark green to red represent increasing diversity. FIGURE 17 Argentaria itonis comb.n. (a) male wing venation; (b and c) male dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 18 Argentaria gen.n. representative species. (a) A . clementia comb.n. ; (b) A . clorimena comb.n. ; (c) A . kendalli comb.n. ; (d) A . pagyris comb.n. ; (e) A . quadrina comb.n. ; (f) A . salvini comb.n . Scale bars 1 cm. FIGURE 19 Argentaria itonis comb.n. A-E, male genitalia (dissection KW-21-41), lateral tip abdomen (a), lateral (b) with posterior view juxta, dorsal (c), aedeagus lateral (d) and aedeagus dorsal (e); (f – i), female genitalia (dissection KW-21-60), lateral view exterior tip abdomen (f), ventral view exterior tip abdomen (g), dorsal view interior abdomen (h), corpus bursae perpendicular to signa (i). Scale bars 1 mm. FIGURE 20 Argentaria gen.n. Species diversity mapped on a 2 degree grid. Colours ranging from dark green to red represent increasing diversity. FIGURE 21 Taguaiba drogoni comb.n. (a) male wing venation; (b and c) male dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 22 Taguaiba gen.n. Species diversity mapped on a 2 degree grid. FIGURE 23 Xenovena murryae comb.n. (a) male wing venation; (b and c) male dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 24 Xenovena murrayae comb.n. (a – e) male genitalia, lateral (a), dorsal (b), anterior view (c), aedeagus lateral (d) and aedeagus dorsal (e); (f – h) female genitalia, lateral view exterior tip abdomen (f), ventral view exterior tip abdomen (g), dorsal view interior abdomen (h). Scale bars 1 mm. FIGURE 25 Xenovena murrayae comb.n. Locality records. FIGURE 26 Trico tricolor tricolor comb.n. (a) male wing venation; (b and c) male dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 27 Trico tricolor fulgora comb.n. (a and b) male dorsal (a) and ventral (b); (c and d) female dorsal (c) and ventral (d). Scale bars 1 cm. FIGURE 28 Trico tricolor comb.n. (a and b) male genitalia, lateral (a) with posterior view juxta, aedeagus lateral (b); (c and d) female genitalia, ventral view exterior tip abdomen (c), dorsal view interior abdomen (d). Scale bars 1 mm. FIGURE 29 Trico tricolor comb.n. Locality records. FIGURE 30 Occulta ocnus comb.n. (a) male wing venation; (b and c) male (LEP-10408) dorsal (b) and ventral (c); (d and e) female (ST ocnus ) dorsal (d) and ventral (e), with specimen labels. Scale bars 1 cm. FIGURE 31 Occulta ocnus comb.n. (a and b) male genitalia (dissection SN-19-157), lateral (a) with posterior view juxta, aedeagus lateral (b); (c – e) female genitalia (dissection NHMUK010402850), lateral view exterior tip abdomen (c), ventral view exterior tip abdomen (d), dorsal view interior abdomen (e), corpus bursae perpendicular to signa (f). Scale bars 1 mm. FIGURE 32 Occulta ocnus comb.n. Locality records. FIGURE 33 Deltaya ocypete comb.n. (a) male wing venation; (b and c) male (FLMNH-MGCL-209681) dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 34 Deltaya gen.n. representative species. (a) D . opima comb.n ; (b) D . louisammour comb.n (FLMNH-MGCL-111522); (c) D . andrei comb.n ; (d) D . probata comb.n . Scale bars 1 cm. TABLE 1 Comparison of characters for distinguishing Deltaya gen.n., Modica gen.n. and related genera.

Modica gen.n.	Emeryus	Paryphthimoides	Colombeia	Scriptor	Deltaya gen.n.	Malaveria
Eyes: hair-like setae	Present	Absent	Present	Present	Present	Present	Present
DHW pale pupil dots in ocellus in cell Cu2-Cu1	Visible	Not visible	Variably visible	Variably visible	Not visible	Not visible	Not visible
VFW: prominent dark brown band (umbra) underlying postdiscal ocelli	Yes	No	Yes, somewhat (except almost absent in P . poltys , P . vestigiata )	Yes	Yes	Yes	No or weak umbra
VHW: dark marginal line in tornus	Thin, not broadening	Thin, not broadening	Thin, not broadening (except P . sheba , P . pseudoconfusa )	Marginal line slightly broader throughout wing	Broadening	Broadening	Thin, not broadening
VHW: postdiscal ocelli in cells Cu1- M3 and M3-M2	Double pupils (silver dots or elongate dashes) distinctly ringed with yellow	Where present, double pupils (silver dots) distinctly ringed with yellow	Variable across species, either double pupils (silver dots) distinctly ringed with yellow, or single pupil (an elongate silver smudge) indistinctly ringed with orange	Where present, double pupils (silver dots) distinctly ringed with yellow	Single pupil (an elongate silver smudge) indistinctly ringed with orange	Double pupils (silver dots or elongate dashes) distinctly ringed with yellow	Double pupils (silver dots or elongate dashes) distinctly ringed with yellow
Male genitalia aedeagus: cornuti	Variably present	Present	Present	Absent	Absent	Present (except in D . andrei and D . probata )	Absent
Female genitalia: lamella antevaginalis	No sclerotized lamella antevaginalis	Wrinkled, sclerotized lamella antevaginalis	Sclerotized lamella antevaginalis in some species	Sclerotized lamella antevaginalis	No sclerotized lamella antevaginalis	Sclerotized lamella antevaginalis (except in D . andrei and D . probata )	Sclerotized ‘spike’-like lamella antevaginalis

FIGURE 35 Deltaya ocypete comb.n. A-E, male genitalia (dissection KW-21-66), lateral (a) with posterior view juxta, dorsal (b), aedeagus lateral (c) and aedeagus dorsal (d), vesica everted showing cornuti (e); (f – i) female genitalia (dissection KW-21-67), lateral view exterior tip abdomen with 8th segment retracted (f), lateral view exterior tip abdomen with 8th segment extended (g), ventral view exterior tip abdomen (h), dorsal view interior abdomen (i). Scale bars 1 mm. FIGURE 36 Deltaya gen.n. Species diversity mapped on a 2 degree grid. Colours ranging from dark green to red represent increasing diversity. FIGURE 37 Modica confusa comb.n. (a) male wing venation; (b and c) male (FLMNH-MGCL-269550) dorsal (b) and ventral (c); (d and e) female dorsal (d) and ventral (e). Scale bars 1 cm. FIGURE 38 Modica gen.n. representative species. (a) M . myncea comb.n (neotype); (b) M . fugitiva comb.n (neotype); (c) M . maripa comb.n (MB-1708-13); (d) M . kamel comb.n (KW-081111-67). Scale bars 1 cm. FIGURE 39 Modica confusa comb.n. (a – d) male genitalia (dissection KW-21-64), lateral (a) with posterior view juxta, dorsal (b), aedeagus lateral (c) and aedeagus dorsal (d); (e – h) female genitalia (dissection KW-21-65), lateral view exterior tip abdomen (e), ventral view exterior tip abdomen (f), dorsal view interior abdomen (g), corpus bursae perpendicular to signa (h). Scale bars 1 mm. FIGURE 40 Modica gen.n. Species diversity mapped on a 2 degree grid. Colours ranging from dark green to red represent increasing diversity. Euptychia Hübner, 1818 = Caenoptychia Le Cerf, 1919 = Ristia Gagarin, 1936 alacristata Neild, Nakahara & Fratello, 2014 [ Neild et al. (2014 , Tropical Lepidoptera Research, 24(1): 4-9)] aquila Fratello, Nakahara & Brévignon, 2015 [ Fratello et al. (2015 , Journal of the Lepidopterists' Society, 69(4): 293-306)] atlantica Nakahara & Freitas, 2017 [ Nakahara et al. (2017 , Neotropical Entomology, 46: 302-309)] attenboroughi Neild, Nakahara, Fratello & Le Crom, 2015 [ Neild et al. (2015 , ZooKeys, 541: 87-108)] audacia Brévignon, Fratello & Nakahara, 2015 [ Fratello et al. (2015 , Journal of the Lepidopterists' Society, 69(4): 293-306)] boulleti (Le Cerf, 1919) [ Freitas et al. (2012 , Neotropical Entomology, 41(6): 461-467)] = virgata Joicey & Talbot, 1924 [ Freitas et al. (2012 , Neotropical Entomology, 41(6): 461-467)] = tigrina (Gagarin, 1936) [ Freitas et al. (2012 , Neotropical Entomology, 41(6): 461-467)] cesarense Pulido, Andrade, Peña & Lamas, 2011 [Pulido et al. (2011, Zootaxa, 2906: 43-51)] - obtusa Nakahara, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] - viloriai Andrade, Pulido, Peña & Lamas, 2011 [Pulido et al. (2011, Zootaxa, 2906: 43-51)] efraini Ríos, 2019 [Ríos (2019, Anartia, 29: 49-53)] = similis Henao, 2019 , preocc. (not Butler, 1867 ) [Ríos (2019, Anartia, 29: 49-53)] enyita Nakahara, Lamas & Willmott, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] enyo Butler, 1867 favonius Nakahara, Vega & Willmott, 2016 [ Nakahara et al. (2016 , Zootaxa, 4184: 358-366)] fernandae Nakahara & Willmott, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] fetna Butler, 1870 granatina Nakahara, Le Crom & Hall, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] hannemanni Forster, 1964 insolata Butler & Druce, 1872 = macrophthalma Staudinger, 1876 jesia Butler, 1869 juanjoi Le Crom, Nakahara & Lamas, 2015 [ Nakahara et al. (2015 , Journal of Research on the Lepidoptera, 48: 59-63)] lacandona Warren & Nakahara, 2015 [ Nakahara et al. (2015 , Journal of Research on the Lepidoptera, 48: 51-57)] marceli Brévignon, 2005 [ Brévignon (2005 , Lambillionea, 105(3)(1): 393-404)] - divisa Benmesbah, Costa, Attal & Viloria, 2021 [Costa et al. (2021, Antenor, 8(1): 2-28)] meta Weymer, 1911 mollina (Hübner, [1818]) = mollina (Hübner, 1808) , nom. nud. - suzannae Brévignon, 2005 [ Brévignon (2005 , Lambillionea, 105(3)(1): 393-404)] mollis Staudinger, 1876 neblina Warren & Nakahara, 2015 [ Nakahara et al. (2015 , Journal of Research on the Lepidoptera, 48: 51-57)] neildi Brévignon, 2005 [ Brévignon (2005 , Lambillionea, 105(3)(1): 393-404)] padroni Nakahara, Lamas & Willmott, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] pegasus Nakahara & J. Hall, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] picea Butler, 1867 pillaca Nakahara & Willmott, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] roraima Nakahara, Fratello & Harvey, 2014 [ Nakahara et al. (2014 , Zootaxa, 3881(3): 291-300)] rubrofasciata L.D. Miller & J.Y. Miller, 1988 rufocincta Weymer, 1911 sarah Benmesbah & Viloria, 2019 [Costa et al. (2019, Anartia, 29: 20-48)] sophiae Zacca, Nakahara, Dolibaina & Dias, 2015 [ Neild et al. (2015 , ZooKeys, 541: 87-108)] truncata Nakahara & J. Hall, 2015 [ Nakahara et al. (2015 , Tropical Lepidoptera Research, 25(2): 63-79)] westwoodi Butler, 1867 - muli Brévignon, 2005 [ Brévignon (2005 , Lambillionea, 105(3)(1): 393-404)] woroina Viloria & Benmesbah, 2020 [Costa et al. (2020, Antenor, 7(1): 19-41)]