Transgenerational inheritance of wing development defects in Drosophila melanogaster induced by cadmium.

The transgenerational inheritance of phenotype induced by environmental factors is a new focus in epigenetic research. In this study, Drosophila melanogaster (F0) was cultured in the medium containing cadmium (Cd, 4.5 mg/kg) from eggs to adults, and offspring (F1-F4) were continuously kept in standard medium (without cadmium). The phenotype analysis showed that cadmium induced developmental defects on wings and apoptosis in the wing disc cells of Drosophila (F0). The wing defects were transmitted for at least four generations even without Cd afterwards. And the effect on the mRNA expression of wing development related genes (shg, omb, F-actin, Mekk1) can be maintained for at least two or three generations. More importantly, under cadmium stress, the post-translational modification (PTM) on the histones H3K4me3 in the third instar larvae and ovaries or testes of adult flies increased significantly, while the levels of H3K9me3 and H3K27me3 decreased significantly. The expression of histone methylation related genes (dSet-1, ash1, Lsd1) increased significantly and these changes can be transmitted to offspring from one or two generations in ovaries or testes. These results suggest that the phenotypic defects of wings caused by cadmium can be inherited to the offspring, and this transgenerational inheritance effect may be related to the epigenetic regulation of histone methylation. Therefore, the adaptability of offspring should be considered when evaluating the toxicity and environmental risk of cadmium.

Silence of yki by miR-7 regulates the Hippo pathway.

The Hippo signaling pathway governs organ size via coordinating cell proliferation and apoptosis, and its dysregulation causes congenital diseases and cancers. The homeostasis of Hippo pathway is achieved through multiple post translational modifications. Through Drosophila genetic screening, we found that miRNAs were also involved in Hippo pathway regulation. Here, we showed that overexpression of miR-7 resulted in small wings, which were neutralized by miR-7-sponge (miR-7-sp) co-expression. Mechanistically, miR-7 inhibited the expression of Hippo pathway target genes. Epistatic analyses revealed that miR-7 modulated Hippo pathway through the transcriptional cofactor Yorkie (Yki). Consistently, overexpression of miR-7 decreased Yki protein. We further found a seed sequence of miR-7 in the yki 3'-UTR region. In addition, we discovered that miR-7 was a transcriptional target of Yki. Thus, a negative feedback loop existed for fine tuning Hippo pathway activity. Taken together, our findings uncovered a novel mechanism by which Yki was silenced by miR-7 for Hippo pathway regulation.

Role of Chitin Deacetylase 1 in the Molting and Metamorphosis of the Cigarette Beetle Lasioderma serricorne.

Chitin deacetylases (CDAs) are chitin-modifying enzymes known to play vital roles in insect metamorphosis and development. In this study, we identified and characterized a chitin deacetylase 1 gene (LsCDA1) from the cigarette beetle Lasioderma serricorne. LsCDA1 contains a 1614 bp open reading frame encoding a protein of 537 amino acids that includes domain structures typical of CDAs. LsCDA1 was mainly expressed in the late larval and late pupal stages. In larval tissues, the highest level of LsCDA1 was detected in the integument. The expression of LsCDA1 was induced by 20-hydroxyecdysone (20E) in vivo, and it was significantly suppressed by knocking down the expression of ecdysteroidogenesis genes and 20E signaling genes. RNA interference (RNAi)-aided silencing of LsCDA1 in fifth-instar larvae prevented the larval-pupal molt and caused 75% larval mortality. In the late pupal stage, depletion of LsCDA1 resulted in the inhibition of pupal growth and wing abnormalities, and the expression levels of four wing development-related genes (LsDY, LsWG, LsVG, and LsAP) were dramatically decreased. Meanwhile, the chitin contents of LsCDA1 RNAi beetles were significantly reduced, and expressions of three chitin synthesis pathway genes (LsTRE1, LsUAP1, and LsCHS1) were greatly decreased. The results suggest that LsCDA1 is indispensable for larval-pupal and pupal-adult molts, and that it is a potential target for the RNAi-based control of L. serricorne.

The wing-specific cuticular protein LmACP7 is essential for normal wing morphogenesis in the migratory locust.

Wings are an indispensable structure in many insects for their foraging, courtship, escape from predators, and migration. Cuticular proteins are major components of the insect cuticle and wings, but there is limited information on how cuticular proteins may play an essential role in wing morphogenesis. We identified a wing-specific cuticular protein, LmACP7, which belongs to the RR-2 subfamily of CPR chitin-binding proteins in the migratory locust. LmACP7 was initially produced in epidermal cells and subsequently migrated to the exocuticle at the pre-ecdysial stage in adult wings. Depletion of LmACP7 transcripts by RNA interference markedly reduced its protein amounts, which consequently led to abnormal wing morphogenesis. The deformed wings were curved, wrinkled, and failed to fully expand. We further demonstrated that the deformation was caused by both severe damage of the endocuticle and death of the epidermal cells in the wings. Based on these data, we propose that LmACP7 not only serves as an essential structural protein in the wing but is also required for the integrity of wing epithelial cells. LmACP7 contributes to production of the wing endocuticle and to the morphogenesis of functional wings in the migratory locust.

A Premature Stop Codon in RAF1 Is the Priority Candidate Causative Mutation of the Inherited Chicken Wingless-2 Developmental Syndrome.

The chicken wingless-2 (wg-2) mutation is inherited in an autosomal recessive fashion, and the resulting phenotype in mutant (wg-2/wg-2) individuals is a developmental syndrome characterized by absent wings, truncated legs, craniofacial as well as skin and feather defects, and kidney malformations. Mapping and genotyping established that the mutation resides within 227 kilobases (kb) of chromosome 12 in a wg-2 congenic inbred line. A capture array was designed to target and sequence the candidate region along with flanking DNA in 24 birds from the line. Many point mutations and insertions or deletions were identified, and analysis of the linked variants indicated a point mutation predicted to cause a premature stop codon in the RAF1 gene. Expression studies were conducted inclusive of all genes in the candidate region. Interestingly, RAF1 transcription was elevated, yet the protein was absent in the mutants relative to normal individuals. RAF1 encodes a protein integral to the Ras/Raf/MAPK signaling pathway controlling cellular proliferation, and notably, human RASopathies are developmental syndromes caused by germline mutations in genes of this pathway. Our work indicates RAF1 as the priority candidate causative gene for wg-2 and provides a new animal model to study an important signaling pathway implicated in limb development, as well as RASopathies.

Wing Abnormality in a Wild-Hatched Whooping Crane (Grus americana) Chick from the Nonmigratory Population in Louisiana, USA.

We describe a wing abnormality in a wild-hatched Whooping Crane (Grus americana) chick from the reintroduced Louisiana, US nonmigratory population. Despite its seemingly compromised flight ability, the chick fledged, reached independence, and lived until 13 mo of age. Necropsy revealed an axial malunion near the left carpus likely resulting from trauma.

Incidence of deformities and variation in shape of mentum and wing of Chironomus columbiensis (Diptera, Chironomidae) as tools to assess aquatic contamination.

Constantly, aquatic ecosystems are under pressure by complex mixtures of contaminants whose effects are not always easy to evaluate. Due to this, organisms are sought in which early warning signs may be detected upon the presence of potentially toxic xenobiotic substances. Thereby, the study evaluated the incidence of deformities and other morphometric variations in the mentum and wing of Chironomus columbiensis exposed to water from some of the Colombian Andes affected by mining, agriculture, and cattle raising. Populations of C. columbiensis were subjected throughout their life cycle (24 days) for two generations (F1 and F2). Five treatments were carried out in controlled laboratory conditions (water from the site without impact, site of mining mercury, mining mercury + cyanide, cattle raising, and agriculture) and the respective control (reconstituted water). Thereafter, the percentage of deformities in the mentum was calculated, and for the morphometric analysis 29 landmarks were digitized for the mentum and 12 for the wing. As a result, four types of deformities were registered in the C. columbiensis mentum, like absence of teeth, increased number of teeth, fusion and space between teeth, none of them detected in the individuals from the control. Additionally, the highest incidence of deformity in F1 occurred in the treatment of mining mercury, while for F2 this took place in the treatments of mining mercury + cyanide, cattle raising and agriculture. Differences were also found with respect to the morphometric variations of the mentum and wing of C. columbiensis among the control and the treatments with water from the creeks intervened. The treatments of mining mercury + cyanide and agriculture had the highest morphological variation in the mentum and wing of C. columbiensis. The results suggest that the anthropogenic impacts evaluated generate alterations in the oral apparatus of the larval state of C. columbiensis and in the adult state provoke alterations in the wing shape (increased width and reduced basal area). These deformities may be related to multiple stress factors, among them the xenobiotics metabolized by the organisms under conditions of environmental contamination.

Multiple factors correlating with wing malformations in the population of Parnassius apollo (Lepidoptera: Papilionidae) restituted from a low number of individuals: A mini review.

The Apollo butterfly, Parnassius apollo (Linnaeus), was common in Europe over 100 years ago, but currently it is considered as near threatened. Different conservation programs have promoted the persistence of this species; however, it is still endangered. An example of such programs was the action devoted to reestablish the Apollo butterfly population in Pieniny National Park (Poland) from only 20-30 individuals which had survived till the last decade of the 20th century. This reintroduction has been successful; however, unexpected developmental problems appeared. Butterflies with deformed or reduced wings became frequent in the population living in the natural habitat, and particularly among those reared under seminatural conditions (in the same environment, but fenced by a net). Until recently, reasons for these malformations remained unknown. However, reports published during last months indicated that there are genetic, biochemical, and microbiological factors contributing to this phenomenon. In the malformed individuals, lesions in the wingless gene and dysfunctions of laccase 1 and 2 were found to be significantly more frequent than in normal insects. A large fraction of butterflies with deformed or reduced wings was devoid of the prokaryotic symbiont Wolbachia, which was present in most normal individuals. Moreover, Yersinia pseudotuberculosis (Pfeiffer) Smith and Thal, and Serratia sp., bacteria pathogenic to insects, were detected in the biological material from both normal and malformed butterflies from this population. These findings are summarized and discussed in this review, in the light of conservation of insects and restitution of their populations from a low number of individuals.

Bisphenol A induced oxidative stress mediated genotoxicity in Drosophila melanogaster.

This study investigates Bisphenol A (BPA) induced oxidative stress that mediates the genotoxicity in in vivo model Drosophila melanogaster. The calculated LC50 for BPA was 12.35 µg/mL. The strains of D. melanogaster were reared in 0.1, 1.0, 2.5 and 5.0 µg/mL BPA treated food media from the embryonic stage (egg); oxidative stress and genotoxicity parameters were analyzed. Food intake analysis confirmed that BPA is not an anti feedant for Drosophila larvae and it consumed BPA containing food. Increased reactive oxygen species (ROS) and lipid peroxidation (LPO) and depletion of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and glutathione-s-transferase (GST) antioxidant activities were observed in BPA treated groups compared to control. Positive single spots/wing frequencies were observed in standard (ST) and high bioactivation (HB) crosses of marker heterozygous (MH; mwh/flr3) and balancer heterozygous (BH; mwh/TM3) genotype flies indicating BPA is mutagenic and not recombinogenic. A significant increase in tail length and % tail DNA in Comet assay after BPA treatment reveals that BPA has a potential to induce the genotoxicity. Present study suggests that BPA exposure induces oxidative stress, which could be one of the possible mechanisms for induction of genotoxicity.

Evidence that the radioprotector effect of ascorbic acid depends on the radiation dose rate.

Many studies have revealed that ascorbic acid (Aa) acts as a powerful inhibitor of genetic damage. The objetive of the present study was to evaluate the radioprotector effect of Aa at two diferent radiation dose rates. The somatic mutation and recombination test in Drosophila melanogaster was used. 48 h larvae were treated for 24 h with 25, 50 and 100 mM of Aa. After pretreatment, larvae were irradiated with 20 Gy of gamma rays administered at 36 or 960 Gy/h. Toxicity, development rate and frequency of mutant spots were recorded. Results provide evidence of a radioprotective effect for all tested concentrations of Aa only when 20 Gy were delivered at 36 Gy/h and only with 25 mM using the 960 Gy/h. To consider the use of Aa as radioprotector or therapeutic agent, it is necessary to know its potential under different situations to avoid unwanted injuries.

Faster Development Covaries with Higher DNA Damage in Grasshoppers (Chorthippus albomarginatus) from Chernobyl.

In Chernobyl, chronic exposure to radioactive contaminants has a variety of deleterious effects on exposed organisms, including genetic damage and mutation accumulation. However, the potential for such effects to be transmitted to the next generation is poorly understood. We captured lesser marsh grasshoppers (Chorthippus albomarginatus) in the Chernobyl Exclusion Zone from sites varying in levels of environmental radiation by more than three orders of magnitude. We then raised their offspring in a common garden experiment in order to assess the effects of parental exposure to radiation on offspring development and DNA damage. Offspring that reached maturity at a younger age had higher levels of DNA damage. Contrary to our hypothesis, parental exposure to radioactive contamination did not affect DNA damage in their offspring possibly because of intervening adaptation or parental compensatory mechanisms. Our results suggest a trade-off between developmental rate and resistance to DNA damage, whereby offspring developing at faster rates do so at the cost of damaging their DNA. This result is consistent with and extends findings in other species, suggesting that faster growth rates cause increased oxidative damage and stress. We propose that growth rates are subject to stabilizing selection balancing the benefits of fast development and the competing need of buffering its damaging effects to macromolecules and tissues.

Toxic effects of the neem oil (Azadirachta indica) formulation on the stink bug predator, Podisus nigrispinus (Heteroptera: Pentatomidae).

This research investigated the effects of neem oil on mortality, survival and malformations of the non-target stink bug predator, Podisus nigrispinus. Neurotoxic and growth inhibitor insecticides were used to compare the lethal and sublethal effects from neem oil on this predator. Six concentrations of neem oil were topically applied onto nymphs and adults of this predator. The mortality rates of third, fourth, and fifth instar nymphs increased with increasing neem oil concentrations, suggesting low toxicity to P. nigrispinus nymphs. Mortality of adults was low, but with sublethal effects of neem products on this predator. The developmental rate of P. nigrispinus decreased with increasing neem oil concentrations. Longevity of fourth instar nymphs varied from 3.74 to 3.05 d, fifth instar from 5.94 to 4.07 d and adult from 16.5 and 15.7 d with 0.5 and 50% neem doses. Podisus nigrispinus presented malformations and increase with neem oil concentrations. The main malformations occur in wings, scutellum and legs of this predator. The neem oil at high and sub lethal doses cause mortality, inhibits growth and survival and results in anomalies on wings and legs of the non-traget predator P. nigrispinus indicating that its use associated with biological control should be carefully evaluated.

Ligand-Bound GeneSwitch Causes Developmental Aberrations in Drosophila that Are Alleviated by the Alternative Oxidase.

Culture of Drosophila expressing the steroid-dependent GeneSwitch transcriptional activator under the control of the ubiquitous α-tubulin promoter was found to produce extensive pupal lethality, as well as a range of dysmorphic adult phenotypes, in the presence of high concentrations of the inducing drug RU486. Prominent among these was cleft thorax, seen previously in flies bearing mutant alleles of the nuclear receptor Ultraspiracle and many other mutants, as well as notched wings, leg malformations, and bristle abnormalities. Neither the α-tubulin-GeneSwitch driver nor the inducing drug on their own produced any of these effects. A second GeneSwitch driver, under the control of the daughterless promoter, which gave much lower and more tissue-restricted transgene expression, exhibited only mild bristle abnormalities in the presence of high levels of RU486. Coexpression of the alternative oxidase (AOX) from Ciona intestinalis produced a substantial shift in the developmental outcome toward a wild-type phenotype, which was dependent on the AOX expression level. Neither an enzymatically inactivated variant of AOX, nor GFP, or the alternative NADH dehydrogenase Ndi1 from yeast gave any such rescue. Users of the GeneSwitch system should be aware of the potential confounding effects of its application in developmental studies.

The RNA binding KH domain of Spoonbill depletes pathogenic non-coding spinocerebellar ataxia 8 transcripts and suppresses neurodegeneration in Drosophila.

Spinocerebellar ataxia 8 (SCA8) pathogenesis is a resultant of gain-of-function machinery that primarily results at the RNA level. It has been reported that expanded non-coding CTG trinucleotide repeat in the ATXN8OS transcripts leads to SCA8 coupled neurodegeneration. Targeted depletion of pathogenic SCA8 transcripts is a viable therapeutic approach. In this report we have focused on the suppression of toxic RNA gain-of-function associated with SCA8. We report suppression of SCA8 associated neurodegeneration by KH RNA binding domain of Spoonbill. KH domain suppresses pathogenic SCA8 associated phenotype in adult flies. Ectopic expression of KH domain leads to massive reduction in the number and size of SCA8 RNA foci. We show that Spoonbill interacts with toxic SCA8 transcripts via its KH domain and promotes its depletion. Till date, no attempts have been made for therapeutic intervention of SCA8 pathogenesis. Further characterization of Spoonbill KH domain may aid us in designing peptide based therapeutics for SCA8 associated neurodegeneration.

Role of the ABC transporter Mdr49 in Hedgehog signaling and germ cell migration.

Coalescence of the embryonic gonad in Drosophila melanogaster requires directed migration of primordial germ cells (PGCs) towards somatic gonadal precursor cells (SGPs). It was recently proposed that the ATP-binding cassette (ABC) transporter Mdr49 functions in the embryonic mesoderm to facilitate the transmission of the PGC attractant from the SGPs; however, the precise molecular identity of the Mdr49-dependent guidance signal remained elusive. Employing the loss- and gain-of-function strategies, we show that Mdr49 is a component of the Hedgehog (hh) pathway and it potentiates the signaling activity. This function is direct because in Mdr49 mutant embryos the Hh ligand is inappropriately sequestered in the hh-expressing cells. Our data also suggest that the role of Mdr49 is to provide cholesterol for the correct processing of the Hh precursor protein. Supporting this conclusion, PGC migration defects in Mdr49 embryos are substantially ameliorated by a cholesterol-rich diet.

Reduced glutathione biosynthesis in Drosophila melanogaster causes neuronal defects linked to copper deficiency.

Glutathione (GSH) is a tripeptide often considered to be the master antioxidant in cells. GSH plays an integral role in cellular redox regulation and is also known to have a role in mammalian copper homeostasis. In vitro evidence suggests that GSH is involved in copper uptake, sequestration and efflux. This study was undertaken to further investigate the roles that GSH plays in neuronal copper homeostasis in vivo, using the model organism Drosophila melanogaster. RNA interference-mediated knockdown of the Glutamate-cysteine ligase catalytic subunit gene (Gclc) that encodes the rate-limiting enzyme in GSH biosynthesis was utilised to genetically deplete GSH levels. When Gclc was knocked down in all neurons, this caused lethality, which was partially rescued by copper supplementation and was exacerbated by additional knockdown of the copper uptake transporter Ctr1A, or over-expression of the copper efflux transporter ATP7. Furthermore, when Gclc was knocked down in a subset of neuropeptide-producing cells, this resulted in adult progeny with unexpanded wings, a phenotype previously associated with copper dyshomeostasis. In these cells, Gclc suppression caused a decrease in axon branching, a phenotype further enhanced by ATP7 over-expression. Therefore, we conclude that GSH may play an important role in regulating neuronal copper levels and that reduction in GSH may lead to functional copper deficiency in neurons in vivo. We provide genetic evidence that glutathione (GSH) levels influence Cu content or distribution in vivo, in Drosophila neurons. GSH could be required for binding Cu imported by Ctr1A and distributing it to chaperones, such as Mtn, CCS and Atox1. Alternatively, GSH could modify the copper-binding and transport activities of Atox1 and the ATP7 efflux protein via glutathionylation of copper-binding cysteines.

Decanalization of wing development accompanied the evolution of large wings in high-altitude Drosophila.

In higher organisms, the phenotypic impacts of potentially harmful or beneficial mutations are often modulated by complex developmental networks. Stabilizing selection may favor the evolution of developmental canalization--that is, robustness despite perturbation--to insulate development against environmental and genetic variability. In contrast, directional selection acts to alter the developmental process, possibly undermining the molecular mechanisms that buffer a trait's development, but this scenario has not been shown in nature. Here, we examined the developmental consequences of size increase in highland Ethiopian Drosophila melanogaster. Ethiopian inbred strains exhibited much higher frequencies of wing abnormalities than lowland populations, consistent with an elevated susceptibility to the genetic perturbation of inbreeding. We then used mutagenesis to test whether Ethiopian wing development is, indeed, decanalized. Ethiopian strains were far more susceptible to this genetic disruption of development, yielding 26 times more novel wing abnormalities than lowland strains in F2 males. Wing size and developmental perturbability cosegregated in the offspring of between-population crosses, suggesting that genes conferring size differences had undermined developmental buffering mechanisms. Our findings represent the first observation, to our knowledge, of morphological evolution associated with decanalization in the same tissue, underscoring the sensitivity of development to adaptive change.

A lack of Wolbachia-specific DNA in samples from apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) individuals with deformed or reduced wings.

Various insects contain maternally inherited endosymbiotic bacteria which can cause reproductive alterations, modulation of some physiological responses (like immunity, heat shock response, and oxidative stress response), and resistance to viral infections. In butterflies, Wolbachia sp. is the most frequent endosymbiont from this group, occurring in about 30 % of species tested to date. In this report, the presence of Wolbachia-specific DNA has been detected in apollo butterfly (Parnassius apollo). In the isolated population of this insect occurring in Pieniny National Park (Poland), malformed individuals with deformed or reduced wings appear with an exceptionally high frequency. Interestingly, while total DNA isolated from most (about 85 %) normal insects contained Wolbachia-specific sequences detected by PCR, such sequences were absent in a large fraction (70 %) of individuals with deformed wings and in all tested individuals with reduced wings. These results indicate for the first time the correlation between malformation of wings and the absence of Wolbachia sp. in insects. Although the lack of the endosymbiotic bacteria cannot be considered as the sole cause of the deformation or reduction of wings, one might suggest that Wolbachia sp. could play a protective role in the ontogenetic development of apollo butterfly.

Enhanced UV-B radiation during pupal stage reduce body mass and fat content, while increasing deformities, mortality and cell death in female adults of solitary bee Osmia bicornis.

The effects of enhanced UV-B radiation on the oogenesis and morpho-anatomical characteristics of the European solitary red mason bee Osmia bicornis L. (Hymenoptera: Megachilidae) were tested under laboratory conditions. Cocooned females in the pupal stage were exposed directly to different doses (0, 9.24, 12.32, and 24.64 kJ/m(2) /d) of artificial UV-B. Our experiments revealed that enhanced UV-B radiation can reduce body mass and fat body content, cause deformities and increase mortality. Following UV exposure at all 3 different doses, the body mass of bees was all significantly reduced compared to the control, with the highest UV dose causing the largest reduction. Similarly, following UV-B radiation, in treated groups the fat body index decreased and the fat body index was the lowest in the group receiving the highest dose of UV radiation. Mortality and morphological deformities, between untreated and exposed females varied considerably and increased with the dose of UV-B radiation. Morphological deformities were mainly manifested in the wings and mouthparts, and occurred more frequently with an increased dose of UV. Cell death was quantified by the Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay (DNA fragmentation) during early stages of oogenesis of O. bicornis females. The bees, after UV-B exposure exhibited more germarium cells with fragmented DNA. The TUNEL test indicated that in germarium, low doses of UV-B poorly induced the cell death during early development. However, exposure to moderate UV-B dose increased programmed cell death. In females treated with the highest dose of UV-B the vast majority of germarium cells were TUNEL-positive.

Wingless gene cloning and its role in manipulating the wing dimorphism in the white-backed planthopper, Sogatella furcifera.

BACKGROUND: Wingless gene (Wg) plays a fundamental role in regulating the segment polarity and wing imaginal discs of insects. The rice planthoppers have an obvious wing dimorphism, and the long- and short-winged forms exist normally in natural populations. However, the molecular characteristics and functions of Wg in rice planthoppers are poorly understood, and the relationship between expression level of Wg and wing dimorphism has not been clarified. RESULTS: In this study, wingless gene (Wg) was cloned from three species of rice planthopper, Sogatella furcifera, Laodelphgax striatellus and Nilaparvata lugens, and its characteristics and role in determining the wing dimorphism of S. furcifera were explored. The results showed that only three different amino acid residuals encoded by Wg were found between S. furcifera and L. striatellus, but more than 10 residuals in N. lugens were different with L. striatellus and S. furcifera. The sequences of amino acids encoded by Wg showed a high degree of identity between these three species of rice planthopper that belong to the same family, Delphacidae. The macropterous and brachypterous lineages of S. furcifera were established by selection experiment. The Wg mRNA expression levels in nymphs were significantly higher in the macropterous lineage than in the brachypterous lineage of S. furcifera. In macropterous adults, the Wg was expressed mainly in wings and legs, and less in body segments. Ingestion of 100 ng/µL double-stranded RNA of Wg from second instar nymphs led to a significant decrease of expression level of Wg during nymphal stage and of body weight of subsequent adults. Moreover, RNAi of Wg resulted in significantly shorter and deformative wings, including shrunken and unfolded wings. CONCLUSION: Wg has high degree of identity among three species of rice planthopper. Wg is involved in the development and growth of wings in S. furcifera. Expression level of Wg during the nymphal stage manipulates the size and pattern of wings in S. furcifera.