Selective sweeps on novel and introgressed variation shape mimicry loci in a butterfly adaptive radiation.

Natural selection leaves distinct signatures in the genome that can reveal the targets and history of adaptive evolution. By analysing high-coverage genome sequence data from 4 major colour pattern loci sampled from nearly 600 individuals in 53 populations, we show pervasive selection on wing patterns in the Heliconius adaptive radiation. The strongest signatures correspond to loci with the greatest phenotypic effects, consistent with visual selection by predators, and are found in colour patterns with geographically restricted distributions. These recent sweeps are similar between co-mimics and indicate colour pattern turn-over events despite strong stabilising selection. Using simulations, we compare sweep signatures expected under classic hard sweeps with those resulting from adaptive introgression, an important aspect of mimicry evolution in Heliconius butterflies. Simulated recipient populations show a distinct 'volcano' pattern with peaks of increased genetic diversity around the selected target, characteristic of sweeps of introgressed variation and consistent with diversity patterns found in some populations. Our genomic data reveal a surprisingly dynamic history of colour pattern selection and co-evolution in this adaptive radiation.

Complex meristematic activity induced by Eucecidoses minutanus on Schinus engleri turns shoots into galls.

PREMISE: Gall-inducing organisms change the development of their host plant organs, resulting in ontogenetic patterns not observed in the non-galled plants. Distinct taxa induce galls on Schinus spp., manipulating meristematic patterns in the host plant in distinct ways. Here we report ontogenetic novelties induced in the lateral buds of S. engleri by Eucecidoses minutanus, a Cecidosidae, whose galls have been poorly understood. METHODS: The anatomy, histochemistry, and histometry of galls in distinct phases of development, non-galled buds, and stems of Schinus engleri were analyzed in parallel with the instars of E. minutanus to detail the morphogenetic changes in the host with each larval stage. RESULTS: Ontogenetic phases of the galls were intricately associated with larval development. First and second-instar larvae induced pericycle and pith cells to dedifferentiate into the gall inner meristem, where hyperplasia and cell hypertrophy characterized the growth and development phase of the gall. The innermost layers were lipid-rich nutritive cells that lined the larval chamber. Additional vascular bundle rows were produced in young galls. Third and fourth instar-larvae were associated with the gall maturation phase: centripetal lignification of the outer parenchyma cell layers, epidermal stratification, and activation of a cambium-like meristem (CLM). The CLM activity resulted in new layers of nutritive cells that differentiated inward as the first layers of nutritive cells were consumed by E. minutanus larvae, and, also, in more parenchyma cell layers that formed outward. All tissues between the innermost layer of nutritive tissue that surround the gall chamber and the outermost layer of the dermal system that externally covers the gall form the gall wall, and increased in thickness until the end of gall maturation. CONCLUSIONS: E. minutanus induces a structurally complex globoid stem gall, modifying all host plant tissues and stimulating a novel meristematic pattern in S. engleri. The gall developmental stages are each related to specific gall-inducing instars, as gall development progresses according to the development of E. minutanus.

Developmental pathway from leaves to galls induced by a sap-feeding insect on Schinus polygamus (Cav.) Cabrera (Anacardiaceae).

Galling sap-feeding insects are presumed to cause only minor changes in host plant tissues, because they usually do not require development of nutritive tissues for their own use. This premise was examined through comparison of the histometry, cytometry and anatomical development of non-galled leaves and galls of Calophya duvauae (Scott) (Hemiptera: Calophyidae) on Schinus polygamus (Cav.) Cabrera (Anacardiaceae). Cell fates changed from non-galled leaves to galls during the course of tissue differentiation. C. duvauae caused changes in dermal, ground, and vascular systems of the leaves of S. polygamus. Its feeding activity induced the homogenization of the parenchyma, and the neoformation of vascular bundles and trichomes. The histometric and cytometric data revealed compensatory effects of hyperplasia and cell hypertrophy in the epidermis, with hyperplasia predominating in the adaxial epidermis. There was a balance between these processes in the other tissues. Thus, we found major differences between the developmental pathways of non-galled leaves and galls. These changes were associated with phenotypic alterations related to shelter and appropriate microenvironmental conditions for the gall inducer. The nondifferentiation of a typical nutritive tissue in this case was compared to other non-phylogenetically related arthropod gall systems, and is suggested to result from convergence associated with the piercing feeding apparatus of the corresponding gall-inducer.

New species of Oecophoridae (Lepidoptera: Gelechioidea) from Chile.

Nine new species of Oecophoridae are described from central and southern Chile: Corita morai Urra, Encolia flava Urra, Endrosis apablazai Urra, Gildita versicolora Urra, Glorita nahuelbutensis Urra, Nagehana maulina Urra, Nagehana parvula Urra, Pirquelia vidali Urra, Zulemita chimbarongensis Urra. Diagnostic characteristics, geographic distribution data, photographs of adults and illustrations of the genital structures of males and females are provided.

Atacamaptilia ambrosiavora gen. et sp. nov. (Lepidoptera: Gracillariidae), a leaf miner of Ambrosia cumanensis (Asteraceae) in the Atacama Desert.

Adult and immature stages of Atacamaptilia ambrosiavora Vargas Espinoza-Donoso gen. et sp. nov. (Lepidoptera: Gracillariidae: Acrocercopinae) from the Atacama Desert of northern Chile are described and illustrated. The Neotropical Atacamaptilia Vargas Espinoza-Donoso gen. nov. resembles the Old World genera Melanocercops Kumata Kuroko, 1988 and Phodoryctis Kumata Kuroko, 1988 in wing venation. However, differences in genitalia morphology enable the recognition of Atacamaptilia Vargas Espinoza-Donoso gen. nov. as a different genus. COI sequences support this delimitation, since it is not closely phylogenetically related to these genera. A divergence of 18% (Kimura 2-parameter model) of A. ambrosiavora Vargas Espinoza-Donoso gen. et sp. nov. to its nearest neighbor Leucospilapteryx omissella (Stainton, 1848) was found. The larvae of A. ambrosiavora Vargas Espinoza-Donoso gen. et sp. nov. mine leaves of Ambrosia cumanensis Kunth (Asteraceae).

Description of Porphyrosela arachisella sp. nov. (Lepidoptera: Gracillariidae), the first report of Lithocolletinae for Brazil.

During a recent survey of leaf-mining microlepidoptera in the Cerrado biome, mines of an undescribed Porphyrosela Braun (Lepidoptera: Gracillariidae: Lithocolletinae) were found associated with the forage peanut, Arachis pintoi Krapov. W.C. Greg. (Fabaceae). Consequently, adults, immature stages and the leaf mine of Porphyrosela arachisella sp. nov. are herein described based on light and scanning electron microscopy. A preliminary analysis of DNA barcode sequences including putative members of other lithocolletine species and all BINs (Barcode Index Numbers) available for Porphyrosela supports P. arachisella as an independent cluster, with 8 to 11% divergence. Its nearest neighbour was the cluster formed by three BINs (BOLD: ADT2137, BOLD: AAG1161 and BOLD: ADU9985) that includes specimens from Australia, Vietnam and Bangladesh. This is the first report of a lithocolletine gracillariid for Brazil, and the third species recognized for the genus in the Neotropical region.

A peculiar new species of Dione (Agraulis) Boisduval & Le Conte (Lepidoptera, Nymphalidae, Heliconiinae) associated with Malesherbia Ruiz & Pavón (Passifloraceae) in xeric western slopes of the Andes.

Butterflies associated with xerophytic environments of the Andes have been little studied, and they exhibit high levels of endemism. Herein Dione (Agraulis) dodona Lamas & Farfán, sp. nov. (Nymphalidae; Heliconiinae) is described, distributed on the western slopes of the Andes of Peru and northern Chile, between 800 and 3,000 m elevation. Adults of both sexes, and the immature stages, are described and illustrated based on light and scanning electron microscopy. The immature stages are associated with MalesherbiatenuifoliaD. Don (Passifloraceae) found in xeric environments, representing a new record of this genus as a host plant for the subfamily Heliconiinae. Conspicuous morphological differences are presented for all stages at the generic level. Based on a phylogenetic analysis of the COI barcode mitochondrial gene fragment, D. (A.) dodona Lamas & Farfán, sp. nov. is distinguished as an independent lineage within the Agraulis clade of Dione, with ca. 5% difference to congeneric species.

A contribution to the knowledge of leaf-mining Phyllocnistis Zeller, 1848 associated with Baccharis (Asteraceae), with description of two new species from Peru (Lepidoptera: Gracillariidae).

We provide new information on Neotropical Phyllocnistis Zeller (Gracillariidae) associated with Baccharis (Asteraceae) in the Andes region. Two new species are described, P. canta Cerdea Farfn, n. sp. and P. elongata Cerdea Farfn, n. sp., from the western slopes of the Andes of Peru between 2,000 and 3,000 m elevation, both associated with Baccharis latifolia (R.P.) Pers. The type locality of P. baccharidis Hering is confirmed; immature stages were found at 1,300 m feeding on Baccharis tucumanensis Hook. Arn. in Quebrada las Higueras (Tucumn, Argentina). Four new locality records for P. furcata Vargas Cerdea are provided, extending its distribution by 450 km to the north of the type locality.

Host Plant Variation and Lack of Genetic Differentiation in Populations of Dione (Agraulis) dodona Lamas & Farfán (Lepidoptera: Nymphalidae).

Dione (Agraulis) dodona (Nymphalidae: Heliconiinae) is a butterfly restricted to the western slopes of the Andes of Peru and Chile and is associated with Malesherbia tenuifolia in xerophytic environments. In this study, we found six additional species of host plants for D. (A.) dodona belonging to the genus Malesherbia (Passifloraceae). We used mitochondrial DNA sequences (COI) and microsatellites to screen genetic variation and investigate population structure in six geographic disjointed populations of D. (A.) dodona associated with distinct Malesherbia species. The PCoA analysis based on allele frequencies evidenced a lack of differentiation among populations and a low FST. The Bayesian cluster analyses revealed the existence of three genetically distinct groups, but almost all individuals present an admixture ancestry. An absence of isolation by distance pattern was observed. Possible scenarios are discussed: a bottleneck or recent colonization from interconnected populations from the south, and ongoing gene flow among local populations by high dispersal through a landscape formed of isolated populations of Malesherbia.

Phyllocnistis furcata sp. nov.: a new species of leaf-miner associated with Baccharis (Asteraceae) from Southern Peru (Lepidoptera, Gracillariidae).

The southwestern Andes of Peru harbors a hidden taxonomic diversity of Lepidoptera. Here a new leaf-mining species of Gracillariidae (Lepidoptera) is described, Phyllocnistis furcata Vargas & Cerdeña, sp. nov., from a dry Andean valley of southern Peru, at 2400 m above sea level. The morphological aspects of adults (male and female) and the immature stages associated with Baccharis alnifolia Meyen & Walp. (Asteraceae) are given, under optical microscopy and scanning electron microscopy. DNA barcodes show that its nearest neighbor is the Atlantic Forest species Phyllocnistis ourea Brito & Moreira, 2017 that feeds on Baccharis anomala DC. The importance of morphological characters from immature stages for diagnosis among congeneric species is also discussed. Phyllocnistis furcata represents the fourth species of Phyllocnistis Zeller for Peru, and first record from the south of Peru for the genus.

A peculiar new species of gall-inducing, clearwing moth (Lepidoptera, Sesiidae) associated with Cayaponia in the Atlantic Forest.

Larvae of most clearwing moths (Lepidoptera, Sesiidae) are endophagous borers of many angiosperms, including their fruits, stems, and roots. Their localized feeding may lead to swellings on those plant parts, but whether the structures produced should be considered true galls is still controversial. In this study we describe a peculiar sesiid moth, Neospheciacecidogena sp. nov. whose larvae induce unusual, external galls on Cayponiapilosa (Vell.) Cogn. (Cucurbitaceae) in the Atlantic Forest of southernmost Brazil. The adults, egg, larva, pupa and the gall are described and illustrated based on light and scanning electron microscopy. Galls are cylindrical and unilocular; they are induced individually on axillary buds of the C.pilosa stem. Unlike larvae of other sesiids, those of N.cecidogena sp. nov. lack abdominal pseudopodia, and show reduced stemata and chaetotaxy. Pupation occurs inside the gall, after having overwintered in the last larval instar. A maximum likelihood tree constructed based on mitochondrial DNA (COI) sequences showed that N.cecidogena sp. nov. is monophyletic and has an average distance of 13% to species of Melittia. The genera Neosphecia Le Cerf, 1916 stat. rev., Premelittia Le Cerf, 1916 stat rev., and Melittina Le Cerf, 1917 stat. rev. are restored from synonyms of Melittia Hübner, 1819 ["1816"].

Spodoptera cosmioides (Lepidoptera: Noctuidae) in Brazil: spatial distribution and relationship in the S. latifascia species group

ABSTRACT Spodoptera cosmioides (Walker, 1858) is an economically relevant polyphagous moth, widely distributed in the Neotropics and part of the Spodoptera latifascia (Walker, 1856) species group. In this study, we used extensive sampling from different regions to describe the spatial distribution of S. cosmioides in Brazil and evaluate its variability both from morphological and molecular perspectives. Variable coloration and several morphological similarities were found among S. cosmioides and congeners of the S. latifascia complex, diverging from each 0.5 to 3.5% of mitochondrial DNA. The genetic divergence at the species level of S. cosmioides was 0.5% throughout Brazil, and a geographic structure was absent, including shared haplotypes with S. descoinsi Lalanne-Cassou & Silvain, 1994. Spodoptera cosmioides was found in all six biomes of Brazil, with the highest abundance recorded in the Cerrado, followed by the Amazon, Atlantic Rainforest, Caatinga, Pantanal, and Pampa.

Vallissiana universitaria (Lepidoptera: Gracillariidae): a new genus and species of leaf-mining moth associated with Erythroxylum (Erythroxylaceae) in the Atlantic Forest of Brazil.

Vallissiana universitaria Pereira Arévalo, a new genus and species of leaf-miner moth (Gracillariidae: Gracillariinae) is described and illustrated with the aid of optical and scanning electron microscopy, including adults, larva, pupa and the mine. Its monophyletic status is confirmed within the subfamily based on a DNA barcode CoI tree. The immature stages are associated with Erythroxylum argentinum O. E. Schulz (Erythroxylaceae) and four larval instars are found, all forming a round blotch mine from the beginning of ontogeny. The first two instars are sap-feeders, using only the epidermal cells, whereas the last two are tissue-feeders, mining the parenchyma cells. Pupation occurs inside the leaf mine within a flimsy, silk-made cocoon. This is the third endemic genus of gracillariid moths described from the Atlantic Forest of Brazil and the first associated with Erythroxylum P. Browne. Characteristics found on the forewing and in the last abdominal segments of the adult were determinant for the proposition of the new genus. The CoI tree indicated that it is closely related to Aspilapteryx, while this genus was recovered as polyphyletic in the analyses. Morphological evidence supports this polyphyly. Consequently, Sabulopteryx Triberti, 1985, stat. nov. is considered a valid genus.

Genomic architecture and introgression shape a butterfly radiation.

We used 20 de novo genome assemblies to probe the speciation history and architecture of gene flow in rapidly radiating Heliconius butterflies. Our tests to distinguish incomplete lineage sorting from introgression indicate that gene flow has obscured several ancient phylogenetic relationships in this group over large swathes of the genome. Introgressed loci are underrepresented in low-recombination and gene-rich regions, consistent with the purging of foreign alleles more tightly linked to incompatibility loci. Here, we identify a hitherto unknown inversion that traps a color pattern switch locus. We infer that this inversion was transferred between lineages by introgression and is convergent with a similar rearrangement in another part of the genus. These multiple de novo genome sequences enable improved understanding of the importance of introgression and selective processes in adaptive radiation.

Feeding preference of Holymenia clavigera and Anisoscelis foliacea marginella (Hemiptera: Coreidae: Anisoscelini) in relation to intra- and interspecific features of host plants (Passifloraceae).

The interactions of Holymenia clavigera (Herbst) and Anisoscelis foliacea marginella (Dallas) with their hosts (Passifloraceae) in southern Brazil are poorly understood. Previous studies have indicated that both species use several host species and plant parts and have superior performance on wild hosts. This study evaluated feeding preferences and host plant use under both laboratory and field conditions. Choice tests were performed to determine preference for Passiflora suberosa parts, preference for P. suberosa and Passiflora misera immature and ripe fruits, and preference for P. suberosa, P. misera, and Passiflora edulis shoots. A field survey was carried out to assess H. clavigera and A. foliacea marginella use of P. suberosa parts. In addition, immature and ripe fruits of P. suberosa and P. misera were characterized in relation to pH, phenols, and anthocyanins. First-instar nymphs preferred the terminal buds, shifting to immature fruits in the second instar. Both coreids more frequently fed on immature fruits than on ripe ones. Neither species showed a significant preference among host species. The immature fruits presented higher phenol content as compared with the ripe fruits; the opposite was observed for anthocyanins. In the field, immature fruits and mature leaves were selected for feeding and resting. Thus, this study confirms the food mixing condition of these coreids, as well as the importance of fruits for their nutrition. Phenols, and presumably water, in the fruits may play a role in their feeding choices.

Overlooked gall-inducing moths revisited, with the description of Andescecidiumparrai gen. et sp. n. and Olierasaizi sp. n. from Chile (Lepidoptera, Cecidosidae).

There are still many gall systems associated with larvae of Lepidoptera in which the true gall-inducers have not been identified to species. Reports on misidentification of gall inducers have been recurrent for these galls, particularly in complex gall-systems that may include inquilines, kleptoparasites, and cecidophages, among other feeding guilds such as predators and parasitoid wasps. Here we describe and illustrate the adults, larvae, pupae and galls, based on light and scanning microscopy, of Andescecidiumparrai gen. et sp. n. and Olierasaizi sp. n., two sympatric cecidosid moths that are associated with Schinuspolygamus (Cav.) Cabrera (Anacardiaceae) in central Chile. Adults, immatures, and galls of the former did not conform to any known cecidosid genus. Galls of A.parrai are external, spherical, and conspicuous, being known for more than one century. However, their induction has been mistakenly associated with either unidentified Coleoptera (original description) or Olieraargentinana Brèthes (recently), a distinct cecidosid species with distribution restricted to the eastern Andes. Galls of O.saizi had been undetected, as they are inconspicuous. They occur under the bark within swollen stems, and may occur on the same plant, adjacent to those of A.parrai. We also propose a time-calibrated phylogeny using sequences from mitochondrial and nuclear loci, including specimens of the new proposed taxa. Thus in addition to clarifying the taxonomy of the Chilean cecidosid species we also tested their monophyly in comparison to congeneric species and putative specimens of all genera of Neotropical and African cecidosids.

Phylogeography of the gall-inducing micromoth Eucecidoses minutanus Brèthes (Cecidosidae) reveals lineage diversification associated with the Neotropical Peripampasic Orogenic Arc.

We investigated the molecular phylogenetic divergence and historical biogeography of the gall-inducing micromoth Eucecidoses minutanus Brèthes (Cecidosidae) in the Neotropical region, which inhabits a wide range and has a particular life history associated with Schinus L. (Anacardiaceae). We characterize patterns of genetic variation based on 2.7 kb of mitochondrial DNA sequences in populations from the Parana Forest, Araucaria Forest, Pampean, Chacoan and Monte provinces. We found that the distribution pattern coincides with the Peripampasic orogenic arc, with most populations occurring in the mountainous areas located east of the Andes and on the Atlantic coast. The phylogeny revealed a marked geographically structured differentiation, which highlights a first split into two major clades: western (Monte and Chacoan) and eastern (Pampean and coastal forests). Together with AMOVA and network analysis, phylogeny revealed the existence of six well-defined lineages, which are isolated by distance. The TMRCA for Eucecidoses was estimated at ca. 65 Mya, and the divergence among major clades occurred by the Plio-Pleistocene ca. 20-25 Mya, with the extant six lineages emerging about 0.9 to 5.7 Mya (later than the rise of Schinus). These results are associated with a diversification pattern of either a late burst of speciation or early extinction. Population range expansion for some lineages concurring with major climatic changes that occurred during the wet-dry events of the Pleistocene in the region was recovered in both neutrality tests and past dynamics through time analysis. A possible biogeographic scenario reconstructed suggests that Eucecidoses likely emerged from a central meta-population in the south and later dispersed (ca. 38 Mya) using western and eastern as two major routes. Thus, a combination of dispersal and vicariance events that occurred in the ancestral populations might have shaped the current distribution of extant lineages. Speciation driven by host plant shift is potentially involved in the evolutionary history of Eucecidoses.

Subtle variation in size and shape of the whole forewing and the red band among co-mimics revealed by geometric morphometric analysis in Heliconius butterflies.

Heliconius are unpalatable butterflies that exhibit remarkable intra- and interspecific variation in wing color pattern, specifically warning coloration. Species that have converged on the same pattern are often clustered in Müllerian mimicry rings. Overall, wing color patterns are nearly identical among co-mimics. However, fine-scale differences exist, indicating that factors in addition to natural selection may underlie wing phenotype. Here, we investigate differences in shape and size of the forewing and the red band in the Heliconius postman mimicry ring (H. erato phyllis and the co-mimics H. besckei, H. melpomene burchelli, and H. melpomene nanna) using a landmark-based approach. If phenotypic evolution is driven entirely by predation pressure, we expect nonsignificant differences among co-mimics in terms of wing shape. Also, a reinforcement of wing pattern (i.e., greater similarity) could occur when co-mimics are in sympatry. We also examined variation in the red forewing band because this trait is critical for both mimicry and sexual communication. Morphometric results revealed significant but small differences among species, particularly in the shape of the forewing of co-mimics. Although we did not observe greater similarity when co-mimics were in sympatry, nearly identical patterns provided evidence of convergence for mimicry. In contrast, mimetic pairs could be distinguished based on the shape (but not the size) of the red band, suggesting an "advergence" process. In addition, sexual dimorphism in the red band shape (but not size) was found for all lineages. Thus, we infer that natural selection due to predation by birds might not be the only mechanism responsible for variation in color patterns, and sexual selection could be an important driver of wing phenotypic evolution in this mimicry ring.

Divergent genetic mechanism leads to spiny hair in rodents.

Spines, or modified hairs, have evolved multiple times in mammals, particularly in rodents. In this study, we investigated the evolution of spines in six rodent families. We first measured and compared the morphology and physical properties of hairs between paired spiny and non-spiny sister lineages. We found two distinct hair morphologies had evolved repeatedly in spiny rodents: hairs with a grooved cross-section and a second near cylindrical form. Compared to the ancestral elliptical-shaped hairs, spiny hairs had higher tension and stiffness, and overall, hairs with similar morphology had similar functional properties. To examine the genetic basis of this convergent evolution, we tested whether a single amino acid change (V370A) in the Ectodysplasin A receptor (Edar) gene is associated with spiny hair, as this substitution causes thicker and straighter hair in East Asian human populations. We found that most mammals have the common amino acid valine at position 370, but two species, the kangaroo rat (non-spiny) and spiny pocket mouse (spiny), have an isoleucine. Importantly, none of the variants we identified are associated with differences in rodent hair morphology. Thus, the specific Edar mutation associated with variation in human hair does not seem to play a role in modifying hairs in wild rodents, suggesting that different mutations in Edar and/or other genes are responsible for variation in the spiny hair phenotypes we observed within rodents.

Comparative morphology of the prothoracic leg in heliconian butterflies: Tracing size allometry, podite fusions and losses in ontogeny and phylogeny.

Prothoracic legs of heliconian butterflies (Nymphalidae, Heliconiinae, Heliconiini) are reduced in size compared to mesothoracic and metathoracic legs. They have no apparent function in males, but are used by females for drumming on host plants, a behavior related to oviposition site selection. Here, taking into account all recognized lineages of heliconian butterflies, we described their tarsi using optical and scanning electron microscopy and searched for podite fusions and losses, and analyzed allometry at the static, ontogenetic and phylogenetic levels. Female tarsi were similar, club-shaped, showing from four to five tarsomeres, each bearing sensilla chaetica and trichodea. Male tarsi were cylindrical, formed from five (early diverging lineages) to one (descendant lineages) either partially or totally fused tarsomeres, all deprived of sensilla. Pretarsi were reduced in both sexes, in some species being either vestigial or absent. Tarsal lengths were smaller for males in almost all species. An abrupt decrease in size was detected for the prothoracic legs during molting to the last larval instar at both histological and morphometric levels. In both sexes, most allometric coefficients found at the population level for the prothoracic legs were negative compared to the mesothoracic leg and also to wings. Prothoracic tarsi decreased proportionally in size over evolutionary time; the largest and smallest values being found for nodes of the oldest and youngest lineages, respectively. Our results demonstrate that evolution of the prothoracic leg in heliconian butterflies has been based on losses and fusions of podites, in association with negative size allometry at static, ontogenetic and phylogenetic levels. These processes have been more pronounced in males. Our study provided further support to the hypothesis that evolution of these leg structures is driven by females, by changing their use from walking to drumming during oviposition site selection. In males the leg would have been selected against due to absence of function and thus progressively reduced in size, in association with podites fusions and lost.