Shorter Wings Research Articles

A Novel Family of Winged-Helix Single-Stranded DNA-Binding Proteins from Archaea

The winged helix superfamily comprises a large number of structurally related nucleic acid-binding proteins. While these proteins are often shown to bind dsDNA, few are known to bind ssDNA. Here, we report the identification and characterization of Sul7s, a novel winged-helix single-stranded DNA binding protein family highly conserved in Sulfolobaceae. Sul7s from Sulfolobus islandicus binds ssDNA with an affinity approximately 15-fold higher than that for dsDNA in vitro. It prefers binding oligo(dT)30 over oligo(dC)30 or a dG-rich 30-nt oligonucleotide, and barely binds oligo(dA)30. Further, binding by Sul7s inhibits DNA strand annealing, but shows little effect on the melting temperature of DNA duplexes. The solution structure of Sul7s determined by NMR shows a winged helix-turn-helix fold, consisting of three α-helices, three β-strands, and two short wings. It interacts with ssDNA via a large positively charged binding surface, presumably resulting in ssDNA deformation. Our results shed significant light on not only non-OB fold single-stranded DNA binding proteins in Archaea, but also the divergence of the winged-helix proteins in both function and structure during evolution.

Selection on dispersal drives evolution of metabolic capacities for energy production in female wing-polymorphic sand field crickets, Gryllus firmus.

Life history and metabolism covary, but the mechanisms and individual traits responsible for these linkages remain unresolved. Dispersal capability is a critical component of life history that is constrained by metabolic capacities for energy production. Conflicting relationships between metabolism and life histories may be explained by accounting for variation in dispersal and maximal metabolic rates. We used female wing‐polymorphic sand field crickets, Gryllus firmus, selected either for long wings (LW, flight‐capable) or short wings (SW, flightless) to test the hypothesis that selection on dispersal capability drives the evolution of metabolic capacities. While resting metabolic rates were similar, long‐winged crickets reached higher maximal metabolic rates than short‐winged crickets, resulting in improved running performance. We further provided insight into the mechanisms responsible for covariation between life history and metabolism by comparing mitochondrial content of tissues involved in powering locomotion and assessing the function of mitochondria isolated from long‐ and short‐winged crickets. Our results demonstrated that larger metabolic capacities in long‐winged crickets were underpinned by increases in mitochondrial content of dorsoventral flight muscle and enhanced bioenergetic capacities of mitochondria within the fat body, a tissue responsible for fuel storage and mobilization. Thus, selection on flight capability correlates with increases in maximal, but not resting metabolic rates, through modifications of tissues powering locomotion at the cellular and organelle levels. This allows organisms to meet high energetic demands of activity for life history. Dispersal capability should therefore explicitly be considered as a potential factor driving the evolution of metabolic capacities.

A new genus and species of gall midges the tribe Winnertziini (Diptera, Cecidomyiidae, Porricondylinae) from lower Eocene Fushun amber from China

The new taxon, Fushuniola mai Fedotova & Perkovsky, gen. et sp. nov., is described from a single female from the lower Eocene (Ypresian) Fushun amber from China. This is the second record of Cecidomyiidae known from Fushun amber. The first record is Cecidomyia bujunensis Naora. The new species is characterized by possessing flagellomeres with short neck and hair-shaped sensoria, very long mouth parts and palpi, wide head, short wings and very long apical lobes of ovipositor. It is distinctly different from all known Winnertziini by the very recurved vein R5 and very wide cells between veins C and R5. A diversity of gall midges from lower Eocene amber is presented. The new genus is close to the extant genus Rhipidoxylomyia, known from upper Eocene Rovno amber.

Resistance of Lepidopteran Egg Parasitoids, Trichogramma japonicum and Trichogramma chilonis, to Insecticides Used for Control of Rice Planthoppers.

Trichogramma wasps are commonly used as biocontrol agents to manage lepidopteran rice pests in rice fields. However, lepidopteran pests synergistically occur with rice planthoppers which are not targeted by Trichogramma. The use of Trichogramma parasitoids in field-based pest control efforts is greatly affected by the application of insecticides targeting planthoppers. As such, insecticide-resistant strains of Trichogramma are urgently needed for the incorporation of these beneficial natural enemies into integrated pest management programs in rice agroecosystems. In the present study, Trichogramma japonicum Ahmead (Hymenoptera: Trichogrammitidae) and Trichogramma chilonis Ishii (Hymenoptera: Trichogrammitidae) were treated with sublethal doses of four insecticides which target rice planthoppers, to generate tolerant strains in the laboratory. The resistance rate of T. japonicum to imidacloprid was the highest (17.8-folds) after 10 successive treatments and experienced 2.5, 4.72, and 7.41-fold increases in tolerance to thiamethoxam, buprofezin, and nitenpyram, respectively. Tolerance of T. chilonis to imidacloprid, thiamethoxam, buprofezin, and nitenpyram were 8.8, 6.9, 4.43, and 5.67-fold greater, respectively. The emergence and deformity (without spreading wings or short wings) rates of T. japonicum and T. chilonis gradually recovered with an increased exposure time of treatments. The fecundity of T. japonicum treated with thiamethoxam was significantly higher than that of the control and T. chilonis treated with thiamethoxam and nitenpyra. Our results demonstrate that screening for insecticide-tolerant/resistant Trichogramma strains was feasible, especially in the pairing of T. japonicum and imidacloprid, which could provide a valuable biological control tool that can be combined with traditional chemical control strategies for use in IPM of rice agroecosystems.

A new species of the genus Rhaphidosoma Amyot et Serville, 1843 (Heteroptera, Reduviidae), with data on its chromosome complement.

A new species, Rhaphidosomapaganicumsp. nov. (Heteroptera: Reduviidae: Harpactorinae: Rhaphidosomatini), is described from the Dry Zone of Myanmar. It is the fifth species of Rhaphidosoma Amyot et Serville, 1843, known from the Oriental Region, and the first record of the genus for Myanmar and Indochina. The structure of the external and internal terminalia of the male and female is described and illustrated in detail. The completely inflated endosoma is described for the first time in reduviids. The complex structure of the ductus seminis is shown; it terminates with a voluminous seminal chamber which opens with a wide secondary gonopore and may be a place where spermatophores are formed. The new species is compared with all congeners from the Oriental Region and Western Asia. It is characterised by the absence of distinct tubercles on the abdominal tergites of the male, the presence only two long tubercles and small rounded ones on the abdominal tergites VII and VI, respectively, in the female, the presence of short fore wing vestiges which are completely hidden under longer fore wing vestiges, and other characters. In addition to the morphological description, an account is given of the male karyotype and the structure of testes of Rh.paganicumsp. nov. and another species of Harpactorinae, Polididusarmatissimus Stål, 1859 (tribe Harpactorini). It was found that Rh.paganicumsp. nov. has a karyotype comprising 12 pairs of autosomes and a multiple sex chromosome system (2n♂=24A+X1X2X3Y), whereas P.armatissimus has a karyotype comprising five pairs of autosomes and a simple sex chromosome system (2n♂=10A+XY). The males of these species were found to have seven and nine follicles per testis, respectively. FISH mapping of 18S ribosomal DNA (major rDNA) revealed hybridisation signals on two of the four sex chromosomes (Y and one of the Xs) in Rh.paganicumsp. nov. and on the largest pair of autosomes in P.armatissimus. The presence of the canonical “insect” (TTAGG)n telomeric repeat was detected in the chromosomes of both species. This is the first application of FISH in the tribe Raphidosomatini and in the genus Polididus Stål, 1858.

Trade-off between flight capability and reproduction in Acridoidea (Insecta: Orthoptera).

In many insect taxa, there is a well‐established trade‐off between flight capability and reproduction. The wing types of Acridoidea exhibit extremely variability from full length to complete loss in many groups, thus, provide a good model for studying the trade‐off between flight and reproduction. In this study, we completed the sampling of 63 Acridoidea species, measured the body length, wing length, body weight, flight muscle weight, testis and ovary weight, and the relative wing length (RWL), relative flight muscle weight (RFW), and gonadosomatic index (GSI) of different species were statistically analyzed. The results showed that there were significant differences in RWL, RFW, and GSI among Acridoidea species with different wing types. RFW of long‐winged species was significantly higher than that of short‐winged and wingless species (p < .01), while GSI of wingless species was higher than that of long‐winged and short‐winged species. The RWL and RFW had a strong positive correlation in species with different wing types (correlation coefficient r = .8344 for male and .7269 for female, and p < .05), while RFW was strong negatively correlated with GSI (r = −.2649 for male and −.5024 for female, and p < .05). For Acridoidea species with wing dimorphism, males with relatively long wings had higher RFW than that of females with relatively short wings, while females had higher GSI. Phylogenetic comparative analysis showed that RWL, RFW, and GSI all had phylogenetic signals and phylogenetic dependence. These results revealed that long‐winged individuals are flight capable at the expense of reproduction, while short‐winged and wingless individuals cannot fly, but has greater reproductive output. The results support the trade‐off between flight and reproduction in Acridoidea.

Wing plastic response to temperature variation in two distantly related NeotropicalDrosophilaspecies (Diptera, Drosophilidae)

Phenotypic plasticity has been described for morphological and life-history traits in many organisms. In Drosophila, temperature drives phenotypic change in several traits, but few Neotropical species have been studied and whether the phenotypic variation associated with plasticity is adaptive remains unclear. Here, we studied the phenotypic response to temperature variation in the distantly related Neotropical species Drosophila mercatorum Patterson and Wheeler, 1942 and Drosophila willistoni Sturtevant, 1916. We evaluate if wing shape variation follows that observed in the Neotropical species Drosophila cardini Sturtevant, 1916: round wings at lower temperatures and narrower wings at higher temperatures. The variation in egg–adult development time and in wing size, shape, and allometry was described using reaction norms and geometric morphometrics. In both species, development time and wing size decreased with increasing temperature and wing allometry showed that size explained ≈10% of the shape variation. Wing shape, however, exhibited contrasting responses. At higher temperatures, D. mercatorum developed slightly slender wings, following the pattern previously found for D. cardini, whereas D. willistoni developed plumper and shorter wings, supporting previous studies on Drosophila melanogaster Meigen, 1830. We conclude that all traits studied here were influenced by temperature, and that wing shape seems also to be influenced by phylogeny.

BODY CONDITION OF BREEDING VERMILION FLYCATCHERS (PYROCEPHALUS RUBINUS) VARIES BETWEEN AREAS WITH AND WITHOUT CATTLE RANCHING IN CENTRAL ARGENTINA

The body condition of birds may be conditioned by intrinsic (e.g., sex, age, migratory and breeding status) and extrinsic factors, such as anthropic activities (e.g., livestock raising). We describe the biometry and body condition of Vermilion Flycatchers (VEFL) according to sex and age and evaluate which factors best predict their body condition during the breeding season in the Espinal of central Argentina. The study area included “Parque Luro” Provincial Reserve and two nearby private ranches with livestock. We captured and banded VEFL using mist nets between October to February, from 2015 to 2019. We calculated the scaled mass indexes, which relates body mass to biometric measures (culmen, tarsus, and tail length, and wing chord) to choose the best indicator of body condition. As explanatory variables, we used the breeding condition of females in one model, breeding condition of males in another model, and in a final model we used date, year and site, and included all individuals. We banded 93 VEFL that did not vary according to age-dependent body size or reproductive condition. Nevertheless, females weighed more, had higher fat levels and shorter wings than males. The body condition was only explained by breeding site, being smaller in one ranch than the other ranch and the reserve. We highlight the importance of reserves and the need for further research to understand the ecological constraints that South American birds face, to promote their conservation.

An exceptional partial skeleton of a new basal raptor (Aves: Accipitridae) from the late Oligocene Namba formation, South Australia

ABSTRACT The Australian pre-Pleistocene fossil record of Accipitridae (eagles, hawks, old-world vultures) comprises one latest Oligocene or early Miocene and one middle Miocene species, each represented by partial bones. Globally, most fossil accipitrids are based on single bones. The recent discovery of an older and considerably more complete accipitrid from late Oligocene sediments in Australia is therefore significant. It is derived from the Pinpa Local Fauna from the Namba Formation at Lake Pinpa, South Australia (~26–24 Ma). The fossil, described as Archaehierax sylvestris gen. et sp. nov., represents a raptor that was larger than the black-breasted buzzard Hamirostra melanosternon but smaller and more gracile than the wedge-tailed eagle Aquila audax. Comprehensive morphological and molecular phylogenetic analyses resolved Archaehierax as a basal accipitrid, not closely related to any living subfamily and perhaps the sister taxon to all other accipitrids exclusive of elanines. Relatively short wings similar to species of Spizaetus and Spilornis suggest it was adapted for flight within enclosed forests. Additional accipitrid fossils from the Namba Formation, a distal femur and a distal humerus, are incomparable with the holotype of A. sylvestris; they may represent distinct species or smaller individuals of the new taxon. lsid:zoobank.org:pub:6A25C569-3E9F-43B8-AAF8-F36CE405C06E

Metric variation in the postcranial skeleton of ostriches,Struthio(Aves: Palaeognathae), with new data on extinct subspecies

AbstractAs a result of numerous fossil and subfossil finds of ostriches, there is great demand for a comprehensive osteometric dataset for the living species and subspecies of the genus Struthio. We meet this demand by providing a set of &amp;gt; 100 measurements for a sample of 18 sexed skeletons, including all living and recently extinct species and subspecies of ostriches. We provide the first mensural data for two extinct subspecies, the hitherto questioned Struthio camelus spatzi from north-western Africa and the Arabian ostrich, Struthio camelus syriacus. The unique skeletal proportions of S. c. spatzi, with a relatively short wing, broad pelvis, short tarsometatarsus and big third toe, confirm the validity of this taxon and suggest an increased stability at the expense of cursoriality, which might have contributed to its extermination by humans. Our biometric analysis of the entire sample suggests a subtle sexual dimorphism in the ostrich skeleton, with females having more robust limb bones (especially wider and/or deeper at the ends) despite being on average smaller than males. If confirmed by further research, this size-independent dimorphism might reflect the independent regulation of the longitudinal and transverse dimensions of bones as revealed by several independent studies of morphological integration (covariance among morphological traits) in the avian skeleton.

Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism

Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

Growth Curves of Ryukyu Scops Owl Nestlings, an Owl Species with Asynchronous Hatching and Reversed Sexual Dimorphism

The growth pattern of an organism is influenced by multiple factors, such as its sex, parental investment, food abundance and sibling competition. For species showing sexual size dimorphism, the sex of siblings is expected to affect the growth of nestlings through differences in competitive hierarchies mediated by an asymmetry in body size. This study describes growth curves of Ryukyu Scops Owl Otus elegans nestlings, based on the analysis of body mass, wing length and tarsus length in 99 individuals. We subsequently tested for effects of sex of the nestlings, having an older male sibling, hatching order and brood size on the growth curves. Overall, we detected significant effects of several predictor variables, although differences between groups were often small. Males and later hatched chicks had lower body mass throughout their growth. The smaller the brood, the heavier the chicks were on days 0–1 and 30–32, although this relationship was reversed on days 8–13. Males and later hatched chicks also had shorter wings. Males had shorter tarsi than females from day 0 to day 4, however, dimorphism in this trait was reversed afterwards. Chicks in a large brood had longer tarsi. Earlier hatched chicks had longer tarsi in the first half of the growth period, but some of this difference had disappeared by the second half. From day 0 to day 3, individuals with an older brother had longer tarsi than those with an older sister. However, halfway through the growth period and thereafter, individuals with an older brother had shorter tarsi than those with an older sister. The fact that older brothers had an effect on the tarsus growth of their siblings was not due to sexual body size asymmetry, as in fact males were smaller than females in wing length and body size. We suggest that sexual differences in aggression mediated by androgens may have played a role.

What does a threatened saproxylic beetle look like? Modelling extinction risk using a new morphological trait database.

The extinction of species is a non-random process, and understanding why some species are more likely to go extinct than others is critical for conservation efforts. Functional trait-based approaches offer a promising tool to achieve this goal. In forests, deadwood-dependent (saproxylic) beetles comprise a major part of threatened species, but analyses of their extinction risk have been hindered by the availability of suitable morphological traits. To better understand the mechanisms underlying extinction in insects, we investigated the relationships between morphological features and the extinction risk of saproxylic beetles. Specifically, we hypothesised that species darker in colour, with a larger and rounder body, a lower mobility, lower sensory perception and more robust mandibles are at higher risk. We first developed a protocol for morphological trait measurements and present a database of 37 traits for 1,157 European saproxylic beetle species. Based on 13 selected, independent traits characterising aspects of colour, body shape, locomotion, sensory perception and foraging, we used a proportional-odds multiple linear mixed-effects model to model the German Red List categories of 744 species as an ordinal index of extinction risk. Six out of 13 traits correlated significantly with extinction risk. Larger species as well as species with a broad and round body had a higher extinction risk than small, slim and flattened species. Species with short wings had a higher extinction risk than those with long wings. On the contrary, extinction risk increased with decreasing wing load and with higher mandibular aspect ratio (shorter and more robust mandibles). Our study provides new insights into how morphological traits, beyond the widely used body size, determine the extinction risk of saproxylic beetles. Moreover, our approach shows that the morphological characteristics of beetles can be comprehensively represented by a selection of 13 traits. We recommend them as a starting point for functional analyses in the rapidly growing field of ecological and conservation studies of deadwood.

Seasonal changes in body size and mass of Red Knots Calidris canutus during autumn migration through southern Baltic

Body size and mass of 1,458 juvenile and 558 adult Knots were measured during autumn migration in Puck Bay, Poland, between 1983 and 1999. The measurements fit well with those gathered along the migration route of Siberian Knots Calidris c. canutus. All linear measurements of adults, except body mass adjusted for size, were significantly higher than those of juveniles. Shorter wing in juveniles than in adults seems to be a general rule regardless of geographical region. Adult females migrated ahead of males, confirmed by decreasing mean measurements. Juveniles carried larger relative energy stores than adults. Puck Bay may be an emergency feeding place for adult Knots, whereas juveniles may use it as one of many stop-over sites. Juveniles were significantly smaller, but had higher adjusted body mass, in late September than earlier, indicating a behavioural difference between late and early birds during preceding stages of migration. Late juveniles probably follow a time-minimising strategy with larger fuel stores and fewer stop-over sites, whereas birds passing in August migrate with very small energetic reserves, making only short flights and stops.

A new fossil species of Procellaria (Aves: Procellariiformes) from the Pliocene of New Zealand

We describe a new Procellaria petrel species from the late Pliocene of Taranaki, New Zealand. The new species is most similar morphologically to the White-Chinned Petrel (P. aequinoctialis), Spectacled Petrel (P. conspicillata) and the Westland Petrel (P. westlandica). Compared with those taxa, the new species has a deeper and shorter premaxilla, longer coracoid and shorter wings, while its legs are a similar size. Today, New Zealand is the centre of global diversity of the genus, with four breeding species. This is the first fossil species of Procellaria to be described from New Zealand, attesting to a reasonably long history of this genus in the region.

Carry‐over effects of environmental stochasticity of the California Current on body condition and wing length of breeding Black‐vented Shearwaters (Puffinus opisthomelas)

Recent climatic variation has led to a change in size or mass in some species. The Black‐vented Shearwater Puffinus opisthomelas is endemic to the California Current System, a highly variable system, giving us cues as to the effects of interannual variability on predators. Here, we report the results of a comparison of biometrics measurement in the short term, 4 years, with different environmental conditions. We found that environmental variability has a direct effect on the body condition of the species, affecting not only body mass but also wing length, with shorter wings as a carry‐over effect of adverse conditions.

Description of two new species of the africana-group of the Genus Gryllotalpa, with a redescription of Gryllotalpa africana Beauvois (Orthoptera: Gryllotalpidae)

Three species of the africana-group of the Genus Gryllotalpa (Orthoptera: Gryllotalpidae) were recorded from Cameroon during an investigation from March 2016 to March 2018: Gryllotalpa africana Beauvois from Buea and Tombel, and two new species, Gryllotalpa tombelensis Simeu-Noutchom and Kekeunou sp. nov. from Tombel and Gryllotalpa babetensis Simeu-Noutchom and Kekeunou sp. nov. from Babété. Gryllotalpa tombelensis is characterised by short wing morphology, vestigial hind wings, small phallic complex, short internal processes, and dorsoventrally flattened body. Gryllotalpa babetensis is distinct from other species of the africana-group by the mesoscutum being more exposed between the pronotum and the base of the fore wings, having a very large phallic complex, with very long internal processes of ectophallus (3 mm long). Both new species were collected in crop fields, but which differ in their habitat characteristics, such as soil moisture, pH and granulometry. A key and comments on the africana-group species of the genus Gryllotalpa and their habitat characteristics are also provided. The finding of the present study aims to help species identification and will help future taxonomists concerned with biodiversity of this group from Cameroon.

Initial analysis of the aerodynamic characteristics of the rotorcraft model

The article presents an aerodynamic analysis of the aircraft models. The process of constructing the physical model began with the development of geometry. Preliminary design was made using SolidWorks program, giving the desired shape with as many details as the method allowed. It had to be made precisely, because later it was to be printed on a 3D printer. In this phase of the project a special hole was also prepared on the model side for mounting in the wind tunnel. The work presents to compare aerodynamic parameters of three models. The models were presented and printed made of thermoplastic material used with a 3D printer. Measurements depend on the determined longitudinal and vertical axial force depending on the angle of attack at the same speed for all objects. The tests were carried out in two types of tunnels: structure 1 in a closed tunnel, structure 2 and 3 in an open tunnel at the Lublin University of Technology. Models 2 and 3 are very close to each other, the only significant difference is the installation of 3 short wings in the place of engine nacelles. The analysis allows you to check whether a given modification has a positive effect on its characteristics.

Butterflies fly using efficient propulsive clap mechanism owing to flexible wings.

Butterflies look like no other flying animal, with unusually short, broad and large wings relative to their body size. Previous studies have suggested butterflies use several unsteady aerodynamic mechanisms to boost force production with upstroke wing clap being a prominent feature. When the wings clap together at the end of upstroke the air between the wings is pressed out, creating a jet, pushing the animal in the opposite direction. Although viewed, for the last 50 years, as a crucial mechanism in insect flight, quantitative aerodynamic measurements of the clap in freely flying animals are lacking. Using quantitative flow measurements behind freely flying butterflies during take-off and a mechanical clapper, we provide aerodynamic performance estimates for the wing clap. We show that flexible butterfly wings, forming a cupped shape during the upstroke and clap, thrust the butterfly forwards, while the downstroke is used for weight support. We further show that flexible wings dramatically increase the useful impulse (+22%) and efficiency (+28%) of the clap compared to rigid wings. Combined, our results suggest butterflies evolved a highly effective clap, which provides a mechanistic hypothesis for their unique wing morphology. Furthermore, our findings could aid the design of man-made flapping drones, boosting propulsive performance.

Identification and Analysis of MicroRNAs Associated with Wing Polyphenism in the Brown Planthopper, Nilaparvata lugens.

Many insects are capable of developing two types of wings (i.e., wing polyphenism) to adapt to various environments. Though the roles of microRNAs (miRNAs) in regulating animal growth and development have been well studied, their potential roles in modulating wing polyphenism remain largely elusive. To identify wing polyphenism-related miRNAs, we isolated small RNAs from 1st to 5th instar nymphs of long-wing (LW) and short-wing (SW) strains of the brown planthopper (BPH), Nilaparvata lugens. Small RNA libraries were then constructed and sequenced, yielding 158 conserved and 96 novel miRNAs. Among these, 122 miRNAs were differentially expressed between the two BPH strains. Specifically, 47, 2, 27 and 41 miRNAs were more highly expressed in the 1st, 3rd, 4th and 5th instars, respectively, of the LW strain compared with the SW strain. In contrast, 47, 3, 29 and 25 miRNAs were more highly expressed in the 1st, 3rd, 4th and 5th instars, respectively, of the SW strain compared with the LW strain. Next, we predicted the targets of these miRNAs and carried out Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. We found that a number of pathways might be involved in wing form determination, such as the insulin, MAPK, mTOR, FoxO and thyroid hormone signaling pathways and the thyroid hormone synthesis pathway. Thirty and 45 differentially expressed miRNAs targeted genes in the insulin signaling and insect hormone biosynthesis pathways, respectively, which are related to wing dimorphism. Among these miRNAs, Nlu-miR-14-3p, Nlu-miR-9a-5p and Nlu-miR-315-5p, were confirmed to interact with insulin receptors (NlInRs) in dual luciferase reporter assays. These discoveries are helpful for understanding the miRNA-mediated regulatory mechanism of wing polyphenism in BPHs and shed new light on how insects respond to environmental cues through developmental plasticity.