A new species of Carvalhomiris from Colombia with an assessment of its phylogenetic position (Heteroptera, Miridae, Orthotylinae)

Abstract Plant bugs, species of Miridae (Heteroptera), are not well known in the Neotropics, and Colombia is not an exception. Based on data from the available systematic catalog (Schuh 2002–2013) fewer than 150 species are recorded from the country, clearly an underestimation. Recent fieldwork has resulted in several new interesting taxa from Colombia. Carvalhomiris Maldonado & Ferreira, 1971, contains three described species from Colombia and Ecuador. From specimens collected in Jardín, Antioquia, Carvalhomirishenryisp. n. is described. Images of the dorsal habitus and the male and female genitalia are provided. Based on morphological examination of the new species and published information, morphological characters were coded to construct a phylogenetic matrix for a cladistic analysis in which the phylogenetic position of the new species is assessed. Carvalhomirishenryisp. n. is the northernmost species of the genus and noteworthy because it is the first record of any species of the genus in the Western Cordillera: all other species are known from the eastern flank of the Andes (Ecuador) or the Eastern Cordillera (Colombia). Natural history observations of the new species, including associations with composites (Asteraceae) are provided. It is speculated that the mirid might be predacious.


Introduction
Miridae are the most diversified family of Heteroptera with more than 11,000 described species (Ferreira et al. 2015, Wheeler and Krimmel 2015, Henry 2017). In the Neotropics, taxonomic efforts to reveal the family's diversity were carried out most notably by José Candido de Melo Carvalho and colleagues (Carvalho and Froeschner 1987, 1990, 1994, Henry and Wheeler 1995. Taxonomic knowledge for the family in the Neotropics is still inadequate (Wheeler 2001), partly because the fauna of certain areas is poorly documented. Colombia, for instance, despite being considered a megadiverse country (Mittermeier 1988, Mittermeier et al. 2011, Arbeláez-Cortés 2013, has only about 150 mirid species listed (Schuh 2002(Schuh -2013, which clearly underestimates the family's actual diversity. Among mirid subfamilies, the Orthotylinae comprise six tribes (Schuh 2002(Schuh -2013, one of which is the paraphyletic Orthotylini (Cassis and Schuh 2012). Within this tribe, Schuh (1974) recognized several informal generic groupings, one of which is the Zanchius group, which includes Carvalhomiris Maldonado & Ferreira, 1971 (Forero 2009). The generic composition of the Zanchius group has varied through time (Schuh 1974, Henry 1995, Forero 2009), but a phylogenetic analysis for the group is not yet available.
Carvalhomiris is characterized by an overall pale yellow coloration with greenish areas, usually strong brachypterism in both sexes, and eyes removed from the anterior margin of the pronotum (Maldonado andFerreira 1971, Forero 2009). The genus has three described species: C. brachypterus Maldonado & Ferreira, 1971 and C. truncatus Forero, 2009, from three localities in Colombia on the Eastern Cordillera, and C. bifurcatus Forero, 2009, from a locality in Ecuador on the eastern flank of the Andes.
Recent fieldwork in Jardín, Antioquia, Colombia, led to the discovery of an additional undescribed species of Carvalhomiris. This new species represents the northernmost distribution of the genus and highlights the wide distributional gap among the known species. Additionally, the discovery of this new species allows the putative synapomorphies for Carvalhomiris proposed by Forero (2009) to be tested. Therefore, we describe this new species of Carvalhomiris, propose the first phylogenetic hypothesis for the genus, and evaluate the phylogenetic position of the new species with respect to previously described species.

Study area and specimens
Specimens were collected near Jardín, Antioquia, Colombia, by beating vegetation along the road. Specimens were mounted, labelled, and deposited in the Entomological collection of the Museo Javeriano de Historia Natural Lorenzo Uribe S.J., of the Pontificia Universidad Javeriana, Bogotá, Colombia (MPUJ_ENT) and in the Entomology Collection of the National Museum of Natural History, Smithsonian Institution, USA (USNM). Quotation marks were used for specimen data cited verbatim. Different labels were separated by a backslash, with comments placed between square brackets.

Dissection and terminology
Genitalia were dissected and examined using a Zeiss Discovery V20 stereoscope or a Nikon SMZ1270 stereoscope following Forero (2008). Digital photographs were taken with a Nikon D5300 attached either to a Nikon SMZ1270 stereoscope or a microscope Nikon Eclipse E100. When taking digital photographs using the Nikon SMZ1270 stereoscope, a modified light dome illumination system (Kawada and Buffington 2016) was used. We used a ring LED illumination system from an Olympus stereoscope.
In proposing a rationale and terminology for the endosomal sclerotizations of male Austromirini, Cassis (2008) designated them as dorsal and ventral endosomal spicules (DES and VES, respectively). Schwartz (2011) expanded the discussion and application of these terms to the Orthotylini, discussing the various configurations of the DES. Schwartz (2011) also noted that several genera of the Orthotylini have a single spiculum that he homologized with the DES. Until further evidence among additional members of the Zanchius group is available, we treated the single spiculum of Carvalhomiris as the DES, despite not completely agreeing with Cassis (2008) that the base of the DES has an expanded keel.

Phylogenetic analyses
Based on the taxonomic revision of Carvalhomiris (Forero 2009) and the documentation of the new species described herein, morphological characters were proposed and coded to build a matrix with 45 characters (Appendix 1, Suppl. material 1). All characters were treated as non-additive. The characters coded document variation among the species relative to body size, antennal structure and coloration, pronotum structure, hemelytron development and structure, and male and female genitalia. The matrix includes seven terminals. The ingroup comprises the three previously described species of Carvalhomiris (Forero 2009) and our species. As outgroups, we included unidentified species of Parachius Distant, 1884 and Itacoris Carvalho, 1947, both members of the Zanchius group (Schuh 1974, Henry 1995, Forero 2009). We also included an additional member of the Orthotylini not closely related to other species of the Zanchius group, an unidentified species of Orthotylus, which also was used to root the trees.
For the phylogenetic analyses, we used parsimony as the optimality criterion. With the number of terminals very small, an exact solution for the matrix can be provided using a branch-and-bound algorithm (Hendy and Penny 1982, Goloboff et al. 2008). Analyses were carried out using TNT version 1.5 Beta (Goloboff and Catalano 2016).  Macropterous male (Figure 1). Similar to brachypterous male in coloration and structure, but with wider pronotum (Table 1), mesoscutum greatly exposed, and forewings fully developed. Brachypterous female (Figure 1). Similar to brachypterous male in coloration and structure, but slightly larger. Thorax: Hemelytron shorter than abdomen, reaching only abdominal segment 5. Genitalia ( Figure 3): Dorsal labiate plate on lateral infoldings with small sclerotized area cephalad of sclerotized infolding ( Figure 3A). Anterior wall with medial margin of gonapophysis 8 asymmetrical ( Figure 3B, arrow), left margin more protuberant than right one, with sclerotized rounded area cephalad of protuberance. Posterior wall as in generic description (Forero 2009), with large, apically expanded interramal dorsal lobes (Fig 3C).
Etymology. We are pleased to dedicate this new species to our friend and colleague Thomas J. Henry. The new name is treated as a Latin noun in the genitive case. Tom is an indefatigable entomologist, always willing to share his knowledge and help others. Tom's enthusiasm for Heteroptera is contagious, not only in the lab but also in the field. Tom collected a long series of specimens of Carvalhomiris in Ecuador in 1996 that was described as C. bifurcatus (Forero 2009); therefore, we take great pleasure in dedicating this new Colombian species of Carvalhomiris to Tom.
Distribution. Known from the type locality on a road near Jardín, Antioquia, in the western Cordillera in Colombia. It was found from 2640 to 2913 meters in eleva-    tion, in a high Andean cloud forest ( Figure 4A). This locality is the northernmost for any species of the genus ( Figure 5).
Plant associations. Specimens of the new species were mostly associated with small roadside herbs of Acmella sp. (Asteraceae) ( Figure 4B). Despite multiple observations, no direct plant feeding was observed.

Discussion
Species of Carvalhomiris are rather homogeneous in external morphology, but their male genitalia are distinct (Forero 2009). Carvalhomiris henryi sp. n. is more closely related to C. brachypterus and C. truncatus, than to C. bifurcatus based on characters of the male and female genitalia (see also phylogenetic analysis below). Carvalhomiris henryi sp. n. is similar to C. truncatus in having a medial process on the posterior margin of the paramere, but this process in C. henryi sp. n. is basally constricted, whereas in C. truncatus it is not. Carvalhomiris henryi sp. n. is similar to C. brachypterus in structure of the left paramere with an acute, medially directed, dorsal process caudad of the sensory area; in C. henryi sp. n. this process is large, not small as in C. brachypterus, and differs from that in C. truncatus which is large and broad. The structure of the dorsal endosomal spicule differentiates C. henryi sp. n. from its congeners. These characters help to identify this new species, but its mixed character associations preclude an unambiguous assessment of phylogenetic relationships.
Heteropteran distributions have been little documented in Colombia, especially for geographically restricted taxa such as species of Carvalhomiris in the high Andes. An increased knowledge of biogeographic patterns might reveal diversification processes that produced these distributional ranges. Unknown is what is driving speciation processes in the Andes, a question that relates to the natural history of Carvalhomiris. It is not certain that species of this genus are strictly phytophagous. They might be omnivores, as is the case in certain orthotylines (Wheeler 2001). To begin answering these questions, further fieldwork is requisite. Because C. henryi sp. n. is known from a different cordillera than its Colombian congeners, the discovery of new species between the known localities can be anticipated. In addition, little-known habitats in Colombia above 2400 m should be thoroughly explored to attain a more complete panorama of the evolution of the group.
The collection of C. henryi sp. n. from a composite, Acmella sp., represents the second plant association for a species of Carvalhomiris. Feeding habits of the new species are unknown, but it might not be strictly phytophagous. The family Tropaeolaceae, on which C. truncatus has been found (Forero 2009), is not closely related to the Asteraceae. Although C. henryi sp. n. was most numerous on Acmella, it also was taken on other plants. This orthotyline might remain on Acmella before dispersing to other plant species when the quality of its typical host deteriorates or when prey numbers decline. The documented associated plants in Carvalhomiris are not closely related. For example, C. truncatus is associated with Tropaeolum sp. (Tropaeolaceae) (Forero 2009), a family far removed from the Asteraceae. Specimens of C. henryi sp. n. were collected on vegetation along the road, and despite being more abundant on Acmella, they were found on other plants in lesser numbers. We suggest that species in Carvalhomiris might be predators, or at least facultatively predaceous. In searching for prey, phytophagous insects can be found readily and in abundance on their host plants. The prey of predatory species, and their associated plants, can vary, which also might be the case in other members of the Zanchius group.

Phylogenetic analysis
The implicit enumeration search strategy for the phylogenetic analysis resulted in two equally parsimonious trees ( Figure 6A; Ci: 81, Ri: 67). The strict consensus showed that either Itacoris or Parachius could be considered the sister group of Carvalhomiris depending on the topology chosen. The strict consensus also showed that Carvalhomiris is monophyletic, but that their internal relationships are not completely resolved. Carvalhomiris henryi sp. n. is placed in a polytomy with C. brachypterus and C. truncatus, with C. bifurcatus as the sister group of this clade. These associations not only show the similarities among the three species, but also point to the particular apomorphies exhibited by C. bifurcatus. Carvalhomiris bifurcatus is also the southernmost species of the genus (Ecuador); it is not known if there is a particular character transformation scheme in relation to geography. The resulting phylogenetic hypotheses do not allow a clear-cut interpretation of a geographic pattern within the C. brachypterus clade. Even though geographically close species also might be expected to be phylogenetically close, the recovered pattern was ambiguous in this respect. The further description of new species of Carvalhomiris might resolve this ambiguity. The monophyly of Carvalhomiris was suggested without the testing of a cladistic hypothesis, but a few putative synapomorphies were noted (Forero 2009). An evaluation of these putative characters in our phylogenetic analysis enables most of the previously suggested synapomorphies to be corroborated: strong brachyptery in both sexes; large genital capsule in males with the anterolateral margin strongly curved; dorsal endosomal spicule C-shaped inserted near the base of the aedeagus; dorsal labiate plate with a median dorsal depression; and lateral infoldings of the sclerotized rings with a caudad acute process. The following were also proposed as putative synapomorphies (Forero 2009), but were not recovered as such in this analysis: right paramere apically expanded, apical margin of paramere serrated, and endosoma with a single spicule.
Further collecting in the wide geographic gap between the known localities of Carvalhomiris species might reveal additional species and allow further testing of phylogenetic relationships in the genus. and from Decanatura de la Facultad de Ciencias de la Pontificia Universidad Javeriana. Al Wheeler kindly provided criticism and suggestions in an earlier version of the manuscript. This paper is a contribution of the project "Actividades docentes y de investigación como apoyo al conocimiento de la biodiversidad colombiana" ID PPTA 00006416 of the Pontificia Universidad Javeriana. Wheeler Table A1.