RESUMO
For insects that exhibit wing polyphenic development, abiotic and biotic signals dictate the adult wing morphology of the insect in an adaptive manner such that in stressful environments the formation of a flight-capable morph is favored and in low-stress environments, a flightless morph is favored. While there is a relatively large amount known about the environmental cues that dictate morph formation in wing polyphenic hemipterans like planthoppers and aphids, whether those cues dictate the same morphs in non-hemipteran (i.e., cricket) wing polyphenic species has not been explicitly investigated. To experimentally test the generality of environmental cue determination of wing polyphenism across taxa with diverse life histories, in this study, we tested the importance of food quantity, parasitic infection, and tactile cues on wing morph determination in the wing polyphenic sand field cricket, Gryllus firmus. Our results also show that certain stress cues, such as severe diet quantity limitation and parasitic infection, actually led to an increase in the production of flightless morph. Based on these findings, our results suggest that physiological and genetic constraints are important to an organism's ability to respond to environmental variation in an adaptive manner beyond simple life history trade-offs.
Assuntos
Gryllidae , Fenótipo , Asas de Animais , Animais , Gryllidae/fisiologia , Gryllidae/anatomia & histologia , Asas de Animais/anatomia & histologia , Características de História de Vida , Meio Ambiente , Masculino , FemininoRESUMO
What limits the size of nature's most extreme structures? For weapons like beetle horns, one possibility is a tradeoff associated with mechanical levers: as the output arm of the lever system-the beetle horn-gets longer, it also gets weaker. This "paradox of the weakening combatant" could offset reproductive advantages of additional increases in weapon size. However, in contemporary populations of most heavily weaponed species, males with the longest weapons also tend to be the strongest, presumably because selection drove the evolution of compensatory changes to these lever systems that ameliorated the force reductions of increased weapon size. Therefore, we test for biomechanical limits by reconstructing the stages of weapon evolution, exploring whether initial increases in weapon length first led to reductions in weapon force generation that were later ameliorated through the evolution of mechanisms of mechanical compensation. We describe phylogeographic relationships among populations of a rhinoceros beetle and show that the "pitchfork" shaped head horn likely increased in length independently in the northern and southern radiations of beetles. Both increases in horn length were associated with dramatic reductions to horn lifting strength-compelling evidence for the paradox of the weakening combatant-and these initial reductions to horn strength were later ameliorated in some populations through reductions to horn length or through increases in head height (the input arm for the horn lever system). Our results reveal an exciting geographic mosaic of weapon size, weapon force, and mechanical compensation, shedding light on larger questions pertaining to the evolution of extreme structures.
Assuntos
Evolução Biológica , Besouros , Cornos , Animais , Masculino , Fenômenos Biomecânicos/fisiologia , Besouros/anatomia & histologia , Besouros/crescimento & desenvolvimento , Besouros/fisiologia , Cornos/anatomia & histologia , Cornos/crescimento & desenvolvimento , Cornos/fisiologia , Remoção , Caracteres Sexuais , JapãoRESUMO
The Japanese rhinoceros beetle Trypoxylus dichotomus is a giant beetle with distinctive exaggerated horns present on the head and prothoracic regions of the male. T. dichotomus has been used as a research model in various fields such as evolutionary developmental biology, ecology, ethology, biomimetics, and drug discovery. In this study, de novo assembly of 615 Mb, representing 80% of the genome estimated by flow cytometry, was obtained using the 10 × Chromium platform. The scaffold N50 length of the genome assembly was 8.02 Mb, with repetitive elements predicted to comprise 49.5% of the assembly. In total, 23,987 protein-coding genes were predicted in the genome. In addition, de novo assembly of the mitochondrial genome yielded a contig of 20,217 bp. We also analyzed the transcriptome by generating 16 RNA-seq libraries from a variety of tissues of both sexes and developmental stages, which allowed us to identify 13 co-expressed gene modules. We focused on the genes related to horn formation and obtained new insights into the evolution of the gene repertoire and sexual dimorphism as exemplified by the sex-specific splicing pattern of the doublesex gene. This genomic information will be an excellent resource for further functional and evolutionary analyses, including the evolutionary origin and genetic regulation of beetle horns and the molecular mechanisms underlying sexual dimorphism.
Assuntos
Besouros , Animais , Feminino , Masculino , Besouros/genética , Fenótipo , Caracteres SexuaisRESUMO
The elaborate ornaments and weapons of sexual selection, such as the vast array of horns observed in scarab beetles, are some of the most striking outcomes of evolution. How these novel traits have arisen, develop, and respond to condition is governed by a complex suite of interactions that require coordination between the environment, whole-animal signals, cell-cell signals, and within-cell signals. Endocrine factors, developmental patterning genes, and sex-specific gene expression have been shown to regulate beetle horn size, shape, and location, yet no overarching mechanism of horn shape has been described. Recent advances in microscopy and computational analyses combined with a functional genetic approach have revealed that patterning genes combined with intricate epithelial folding and movement are responsible for the final shape of a beetle head horn.
Assuntos
Evolução Biológica , Padronização Corporal/genética , Besouros/genética , Cornos/anatomia & histologia , Animais , Besouros/anatomia & histologia , Besouros/crescimento & desenvolvimento , Epitélio/anatomia & histologia , Epitélio/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento/genética , Cornos/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
Thrips tabaci is a key pest of onions, especially in the Pacific Northwestern USA. Management of T. tabaci is dominated by the application of various insecticides. However, T. tabaci is known to develop insecticide resistance which possibly leads to control failures, crop loss, and environmental concern. Here, we evaluated resistance status of T. tabaci populations from conventional and organic commercial onion fields to three widely used insecticides: oxamyl, methomyl, and abamectin with on-field concentration-mortality bioassays. The biochemistry and molecular mechanisms underlying resistance to these insecticides were also investigated by using enzymatic assays and detecting resistance-associated mutations. Field-evolved resistance to oxamyl, methomyl and abamectin were detected in most of the T. tabaci populations collected from conventional onion farms. At the labeled field rate, all the tested insecticides, particularly methomyl and oxamyl, had significantly reduced efficacy. Enzymatic assays of insecticide target and detoxification enzymes indicated that T. tabaci populations in Western USA onions harbor multiple mechanisms of resistance including enhanced activities of detoxification enzymes and target site insensitivity. Our results provide new information in understanding the dynamics of T. tabaci adaptation to multiple insecticides, which will help to design sustainable insecticide resistance management strategies for T. tabaci. Furthermore, this study provides the foundation for future research in identifying the biochemical and molecular markers associated with insecticide resistance in T. tabaci.
Assuntos
Inseticidas , Tisanópteros , Animais , Resistência a Inseticidas , Metomil , CebolasRESUMO
The two-spotted spider mite, Tetranychus urticae, is a polyphagous pest feeding on over 1100 plant species, including numerous highly valued economic crops. The control of T. urticae largely depends on the use of acaricides, which leads to pervasive development of acaricide resistance. Cytochrome P450-mediated metabolic detoxification is one of the major mechanisms of acaricide resistance in T. urticae. NADPH-cytochrome P450 reductase (CPR) plays as a crucial co-factor protein that donates electron(s) to microsomal cytochrome P450s to complete their catalytic cycle. This study seeks to understand the involvement of CPR/P450 in acaricide resistance in T. urticae. The full-length cDNA sequence of T. urticae's CPR (TuCPR) was cloned and characterized. TuCPR was ubiquitously transcribed in different life stages of T. urticae and the highest transcription was observed in the nymph and adult stages. TuCPR was constitutively over-expressed in six acaricide resistant populations compared to a susceptible one. TuCPR transcriptional expression was also induced by multiple acaricides in a time-dependent manner. Down-regulation of TuCPR via RNA interference (RNAi) in T. urticae led to reduced enzymatic activities of TuCPR and cytochrome P450s, as well as a reduction of resistance to multiple acaricides, abamectin, bifenthrin, and fenpyroximate. The outcome of this study highlights CPR as a potential novel target for eco-friendly control of T. urticae and other related plant-feeding pests.
Assuntos
Acaricidas , Tetranychidae , Animais , Sistema Enzimático do Citocromo P-450 , NADPH-Ferri-Hemoproteína Redutase , Interferência de RNARESUMO
Wing polyphenism is a phenomenon in which one genotype can produce two or more distinct wing phenotypes adapted to the particular environment. What remains unknown is how wing pad development is controlled downstream of endocrine signals such as insulin and JNK pathways. We show that genes important in cellular proliferation, cytokinesis, and cell cycle progression are necessary for growth and development of long wings. Wing pad cellular development of the long-winged morph was characterized by a highly structured epithelial layer with microvilli-like structures. Cells of adult short wing pads are largely in the G2/M phase of the cell cycle, whereas those of long wings are largely in G1. Our study is the first to report the comparative developmental and cellular morphology and structure of the wing morphs and to undertake a comprehensive evaluation of the cell cycle genes necessary for wing development of this unique, adaptive life history strategy.
RESUMO
Multiple acaricide resistance in Tetranychus urticae continues to threaten crop production globally, justifying the need to adequately study resistance for sustainable pest management. Most studies on acaricide resistance have focused on the acute contact toxicity of acaricides with little or no information on the behavioral responses elicited after acaricide exposure. Furthermore, the impact of physiological resistance on these behavioral responses remains unknown in most pest species, including T. urticae. We tested the effect of acaricide resistance on contact toxicity, irritancy and repellency of mitochondrial electron transport inhibitor of complex I (MET-I) and mite growth inhibitor (MGI) acaricides on multiple T. urticae strains. We also tested whether acaricides with similar physiological target site/mode of action also elicit similar behavioral effects on T. urticae strains. MET-I acaricides (fenazaquin, fenpyroximate, and pyrabiden) and MGIs (clofentezine, hexythiazox and etoxazole) elicited a dose-dependent irritant and repellent effect on T. urticae. Selection of strains for physiological resistance to these acaricides affected the behavioral response of T. urticae, especially in MET-I resistant strains, that showed reduced irritancy and repellency to MET-I acaricides. Behavioral response also affected the oviposition of T. urticae, where strains generally showed preferential oviposition away from the acaricides. The outcome of this study highlights negative consequences of acaricide resistance that can potentially affect T. urticae management.
Assuntos
Acaricidas/farmacologia , Ácaros/efeitos dos fármacos , Controle de Pragas , Tetranychidae/efeitos dos fármacos , Acaricidas/efeitos adversos , Animais , Clorobenzenos/farmacologia , Humanos , Ácaros/patogenicidade , Oxazóis/farmacologia , Tetranychidae/patogenicidadeRESUMO
Japanese rhinoceros beetle (Trypoxylus dichotomus) males have exaggerated head horns that they use as weapons in combat over reproductive opportunities. In these contests, there is an advantage to having a longer horn, and there seems to be little cost to horn exaggeration. However, populations vary in the amount of horn exaggeration across this widespread species. Here, we examine four populations and quantify scaling and functional morphology of the horn. We then measure force production by the horn system in a combat-relevant movement. We find that not only does horn length vary among populations, but allometry of lever mechanics and force production varies in a complex way. For instance, some beetle populations make relatively long horns, but exert relatively low forces. Other populations make shorter horns and produce higher forces during fights. We suggest that this performance variation could be associated with differences in the intensity or type of sexual selection across the species.
RESUMO
Males of the Asian rhinoceros beetle, Trypoxylus dichotomus, possess exaggerated head and thoracic horns that scale dramatically out of proportion to body size. While RNAi-mediated knockdowns of the insulin receptor suggest that the insulin signaling pathway regulates nutrition-dependent growth including exaggerated horns, the genes that regulate disproportionate growth have yet to be identified. We used RNAi-mediated knockdown of several genes to investigate their potential role in growth and scaling of the sexually dimorphic, exaggerated head horns of T. dichotomus. Knockdown of the insulin signaling substrate chico and the ecdysone response element broad caused significant decreases in head horn length, while having no or minimal effects on other structures such as elytra and tibiae. However, scaling of horns to body size was not affected by either knockdown. In addition, knockdown of phosphatase and tensin homolog, a negative regulator of the insulin signaling pathway, had no significant effects on any trait. Our results do not identify any candidate genes that may specifically mediate the allometric aspect of horn growth, but they do confirm the insulin signaling pathway as a mediator of conditional trait expression, and importantly implicate the ecdysone signaling pathway, possibly in conjunction with insulin signaling, as an additional mediator of horn growth.
Assuntos
Besouros/crescimento & desenvolvimento , Besouros/genética , Proteínas de Insetos/genética , Animais , Ecdisona/metabolismo , Cabeça/crescimento & desenvolvimento , Proteínas de Insetos/metabolismo , Insulina/fisiologia , Masculino , Elementos de Resposta , Transdução de Sinais/genéticaRESUMO
Members of the phylum Arthropoda, comprising over 80% of total animal species, have evolved regenerative abilities, but little is known about the molecular mechanisms mediating this process. Transforming growth factor ß (TGF-ß) signaling mediates a diverse set of essential processes in animals and is a good candidate pathway for regulation of regeneration in arthropods. In this study we investigated the role of activin signaling, a TGF-ß superfamily pathway, in limb regeneration in the crayfish. We identified and cloned a downstream transcription factor in the activin pathway, Smox, and characterized its function with regard to other elements of the activin signaling pathway. Gene knockdown of Smox by RNAi induced regeneration of complete but smaller pereopods after autotomy. This indicates that activin signaling via Smox functions in regulation of pereopod growth and size. The expression levels of both Smox and the activin receptor babo were closely correlated with molting. The expression level of Smox increased when babo was knocked down by RNAi, indicating that Smox and babo transcription are linked. Our study suggests that the Babo-Smox system in activin signaling is conserved in decapods, and supports an evolutionary conservation of this aspect of molecular signaling during regeneration between protostomes and deuterostomes.
Assuntos
Astacoidea/fisiologia , Proteínas Smad Reguladas por Receptor/metabolismo , Animais , Clonagem Molecular , Extremidades/fisiologia , Técnicas de Silenciamento de Genes , Regeneração , Proteínas Smad Reguladas por Receptor/química , Proteínas Smad Reguladas por Receptor/genéticaRESUMO
Among the most dramatic examples of sexual selection are the weapons used in battles between rival males over access to females. As with ornaments of female choice, the most "exaggerated" sexually selected weapons vary from male to male more widely than other body parts (hypervariability), and their growth tends to be more sensitive to nutritional state or physiological condition compared with growth of other body parts ("heightened" conditional expression). Here, we use RNAseq analysis to build on recent work exploring these mechanisms in the exaggerated weapons of beetles, by examining patterns of differential gene expression in exaggerated (head and thorax horns) and non-exaggerated (wings, genitalia) traits in the Asian rhinoceros beetle, Trypoxylus dichotomus. Our results suggest that sexually dimorphic expression of weaponry involves large-scale changes in gene expression, relative to other traits, while nutrition-driven changes in gene expression in these same weapons are less pronounced. However, although fewer genes overall were differentially expressed in high- vs. low-nutrition individuals, the number of differentially expressed genes varied predictably according to a trait's degree of condition dependence (head horn > thorax horn > wings > genitalia). Finally, we observed a high degree of similarity in direction of effects (vectors) for subsets of differentially expressed genes across both sexually dimorphic and nutritionally responsive growth. Our results are consistent with a common set of mechanisms governing sexual size dimorphism and condition dependence.
Assuntos
Estruturas Animais/anatomia & histologia , Besouros/anatomia & histologia , Caracteres Sexuais , Comportamento Sexual Animal , Animais , Besouros/genética , Feminino , Masculino , Análise de Sequência de RNA , TranscriptomaRESUMO
Tetranychus urticae Koch is a generalist pest of economic crops and is notorious for its rapid development of acaricide resistance. This poses a significant threat to the sustainability of integrated pest management (IPM) in cropping systems plagued by T. urticae. It is critical to evaluate the resistance status of T. urticae populations on crops and identify any underlying resistance mechanisms. This study investigated the efficacy of five major acaricides on T. urticae populations on peppermint and silage corn in the Pacific Northwestern United States and identified the underlying resistance mechanisms. Significant variations in acaricide resistance status of T. urticae populations were identified to abamectin, bifenthrin, fenpyroximate, hexythiazox, and spirodiclofen. In most cases, T. urticae populations from silage corn exhibited greater levels of acaricide resistance relative to peppermint populations. We detected known target-site mutations: F1534S and F1538I (conferring resistance to bifenthrin), G126S (linked with resistance to bifenazate), and I1017 (conferring resistance to hexythiazox and etoxazole) in 10, 90, and 90% of the populations, respectively, from peppermint fields. These four mutations were identified in all the populations collected from silage corn fields. Significantly higher transcript levels of metabolic genes associated with resistance to abamectin, fenpyroximate, and spirodiclofen were observed in some T. urticae populations collected from both peppermint and silage corn fields. This study provides evidence of multiple resistance to diverse active ingredients in field populations of T. urticae and the reliability of known molecular markers for active acaricide resistance monitoring. The observed resistance pattern will help in designing a sustainable IPM program for T. urticae.
Assuntos
Acaricidas , Adaptação Fisiológica , Resistência a Inseticidas , Tetranychidae/genética , Animais , Feminino , Mutação PuntualRESUMO
Beetle horns are attractive models for studying the evolution of novel traits, as they display diverse shapes, sizes, and numbers among closely related species within the family Scarabaeidae. Horns radiated prolifically and independently in two distant subfamilies of scarabs, the dung beetles (Scarabaeinae), and the rhinoceros beetles (Dynastinae). However, current knowledge of the mechanisms underlying horn diversification remains limited to a single genus of dung beetles, Onthophagus. Here we unveil 11 horn formation genes in a rhinoceros beetle, Trypoxylus dichotomus. These 11 genes are mostly categorized as larval head- and appendage-patterning genes that also are involved in Onthophagus horn formation, suggesting the same suite of genes was recruited in each lineage during horn evolution. Although our RNAi analyses reveal interesting differences in the functions of a few of these genes, the overwhelming conclusion is that both head and thoracic horns develop similarly in Trypoxylus and Onthophagus, originating in the same developmental regions and deploying similar portions of appendage patterning networks during their growth. Our findings highlight deep parallels in the development of rhinoceros and dung beetle horns, suggesting either that both horn types arose in the common ancestor of all scarabs, a surprising reconstruction of horn evolution that would mean the majority of scarab species (~35,000) actively repress horn growth, or that parallel origins of these extravagant structures resulted from repeated co-option of the same underlying developmental processes.
Assuntos
Besouros/genética , Larva/genética , Animais , Evolução Biológica , Regulação da Expressão Gênica no Desenvolvimento/genética , Cornos/anatomia & histologia , Cornos/embriologia , Fenótipo , Interferência de RNA , Especificidade da EspécieRESUMO
Food quality is a critical environmental condition that impacts an animal's growth and development. Many insects facing this challenge have evolved a phenotypically plastic, adaptive response. For example, many species of insect exhibit facultative wing growth, which reflects a physiological and evolutionary trade-off between dispersal and reproduction, triggered by environmental conditions. What the environmental cues are and how they are transduced to produce these alternative forms, and their associated ecological shift from dispersal to reproduction, remains an important unsolved problem in evolutionary ecology. In this study, we investigated the role that host quality has on the induction of wing development in a wing polyphenic insect exhibiting strong tradeoffs in investment between dispersal and reproduction, the brown planthopper, a serious rice pest in Asia. As rice plants grow, the short-winged brown planthopper dominates the population, but a shift occurs as the plants mature and senesce in the field such that long-winged brown planthoppers emerge and migrate. It remains unknown how changes in the rice plant induce development of the long-winged morph, despite recent discoveries on the role of the insulin and JNK signaling pathways in wing development. We found that by mimicking the glucose concentration of senescing rice plants, we significantly increased the proportion of long-winged female planthoppers. The effects of glucose on wing morph are additive with previously described effects of density. Our results show that host quality both directly regulates phenotypic plasticity and interacts with other factors such as density to produce the appropriate phenotype for specific environmental conditions.
Assuntos
Evolução Biológica , Hemípteros/fisiologia , Interações Hospedeiro-Parasita/fisiologia , Oryza/parasitologia , Asas de Animais/fisiologia , Animais , Feminino , Hemípteros/anatomia & histologia , Masculino , Asas de Animais/anatomia & histologiaRESUMO
Changes in food availability and crowding are two critical environmental conditions that impact an animal's trajectory toward either reproduction or migration. Many insects facing this challenge have evolved wing polymorpisms that allow them to respond to changing conditions. When conditions favor reproduction, wing polymorphic species produce adults that either have no wings or short, non-functional wings; however, when conditions favor migration, adults with functional wings and robust flight muscles develop. Here we review three recently reported signaling pathways regulating wing polyphenism in wing polymorphic crickets, aphids, and brown planthoppers: juvenile horomone/ecdysone signaling, insulin signaling, and Jun-N-terminal Kinase (JNK) signaling. Understanding how these pathways respond to nutrition, stress and crowding with the appropriate adaptive phenotype is an important step in understanding how life-history trade-offs evolve.
Assuntos
Sistema Endócrino/fisiologia , Insetos/fisiologia , Asas de Animais/crescimento & desenvolvimento , Migração Animal/fisiologia , Fenômenos Fisiológicos da Nutrição Animal , Animais , Aglomeração , Fenótipo , Estresse FisiológicoRESUMO
Males of the Asian rhinoceros beetle, Trypoxylus dichotomus, possess exaggerated head and thoracic horns that scale dramatically out of proportion to body size. While studies of insulin signaling suggest that this pathway regulates nutrition-dependent growth including exaggerated horns, what regulates disproportionate growth has yet to be identified. The Fat signaling pathway is a potential candidate for regulating disproportionate growth of sexually-selected traits, a hypothesis we advanced in a previous paper (Gotoh et al., 2015). To investigate the role of Fat signaling in the growth and scaling of the sexually dimorphic, condition-dependent traits of the in the Asian rhinoceros beetle T. dichotomus, we used RNA interference to knock down expression of fat and its co-receptor dachsous. Knockdown of fat, and to a lesser degree dachsous, caused shortening and widening of appendages, including the head and thoracic horns. However, scaling of horns to body size was not affected. Our results show that Fat signaling regulates horn growth in T. dichotomus as it does in appendage growth in other insects. However, we provide evidence that Fat signaling does not mediate the disproportionate, positive allometric growth of horns in T. dichotomus.
Assuntos
Caderinas/metabolismo , Besouros/crescimento & desenvolvimento , Besouros/metabolismo , Caracteres Sexuais , Animais , Caderinas/genética , Besouros/genética , Besouros/ultraestrutura , Técnicas de Silenciamento de Genes , Masculino , Transdução de SinaisRESUMO
One of the defining features of the evolutionary success of insects is the morphological diversification of their appendages, especially mouthparts. Although most insects share a common mouthpart ground plan, there is remarkable diversity in the relative size and shapes of these appendages among different insect lineages. One of the most prominent examples of mouthpart modification can be found in the enlargement of mandibles in stag beetles (Coleoptera, Insecta). In order to understand the proximate mechanisms of mouthpart modification, we investigated the function of appendage-patterning genes in mandibular enlargement during extreme growth of the sexually dimorphic mandibles of the stag beetle Cyclommatus metallifer. Based on knowledge from Drosophila and Tribolium studies, we focused on seven appendage patterning genes (Distal-less (Dll), aristaless (al), dachshund (dac), homothorax (hth), Epidermal growth factor receptor (Egfr), escargot (esg), and Keren (Krn). In order to characterize the developmental function of these genes, we performed functional analyses by using RNA interference (RNAi). Importantly, we found that RNAi knockdown of dac resulted in a significant mandible size reduction in males but not in female mandibles. In addition to reducing the size of mandibles, dac knockdown also resulted in a loss of the serrate teeth structures on the mandibles of males and females. We found that al and hth play a significant role during morphogenesis of the large male-specific inner mandibular tooth. On the other hand, knockdown of the distal selector gene Dll did not affect mandible development, supporting the hypothesis that mandibles likely do not contain the distal-most region of the ancestral appendage and therefore co-option of Dll expression is unlikely to be involved in mandible enlargement in stag beetles. In addition to mandible development, we explored possible roles of these genes in controlling the divergent antennal morphology of Coleoptera.
Assuntos
Padronização Corporal/genética , Besouros/embriologia , Mandíbula/embriologia , Caracteres Sexuais , Animais , Evolução Biológica , Receptores ErbB/fisiologia , Feminino , Proteínas de Insetos/genética , Proteínas de Insetos/fisiologia , Masculino , Processos de Determinação SexualRESUMO
Quantitative real-time PCR (qRT-PCR) is an extensively used, high-throughput method to analyze transcriptional expression of genes of interest. An appropriate normalization strategy with reliable reference genes is required for calculating gene expression across diverse experimental conditions. In this study, we aim to identify the most stable reference genes for expression studies of xenobiotic adaptation in Tetranychus urticae, an extremely polyphagous herbivore causing significant yield reduction of agriculture. We chose eight commonly used housekeeping genes as candidates. The qRT-PCR expression data for these genes were evaluated from seven populations: a susceptible and three acaricide resistant populations feeding on lima beans, and three other susceptible populations which had been shifted host from lima beans to three other plant species. The stability of the candidate reference genes was then assessed using four different algorithms (comparative ΔCt method, geNorm, NormFinder, and BestKeeper). Additionally, we used an online web-based tool (RefFinder) to assign an overall final rank for each candidate gene. Our study found that CycA and Rp49 are best for investigating gene expression in acaricide susceptible and resistant populations. GAPDH, Rp49, and Rpl18 are best for host plant shift studies. And GAPDH and Rp49 were the most stable reference genes when investigating gene expression under changes in both experimental conditions. These results will facilitate research in revealing molecular mechanisms underlying the xenobiotic adaptation of this notorious agricultural pest.
Assuntos
Tetranychidae/metabolismo , Xenobióticos/metabolismo , Algoritmos , Animais , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Crowding and changes in food availability are two critical environmental conditions that impact an animal's trajectory toward either migration or reproduction. Many insects facing this challenge have evolved wing polyphenisms. When conditions favor reproduction, wing polyphenic species produce adults that either have no wings or short, non-functional wings. Facultative wing growth reflects a physiological and evolutionary trade-off between migration and reproduction, triggered by environmental conditions. How environmental cues are transduced to produce these alternative forms, and their associated ecological shift from migration to reproduction, remains an important unsolved problem in evolutionary ecology. The brown planthopper, a wing polymorphic insect exhibiting strong trade-offs in investment between migration and reproduction, is one of the most serious rice pests in Asia. In this study, we investigated the function of four genes in the insulin-signaling pathway known to couple nutrition with growth, PI3 Kinase (PI3K), PDK1, Akt (Protein Kinase B), and the forkhead gene FOXO. Using a combination of RNA interference and pharmacological inhibitor treatment, we show that all four genes contribute to tissue level regulation of wing polymorphic development in this insect. As predicted, silencing of the NlPI3K, NlAkt and NlPDK1 through dsRNA and with the pharmacological inhibitor Perifosine resulted in short-winged brown planthoppers, whereas knockdown of NlFOXO resulted in long-winged planthoppers. Morphometric analyses confirm that phenotypes from our manipulations mimic what would be found in nature, i.e., major parameters such as bristle number, wing area and body weight are not significantly different from non-experimental animals. Taken together, these data implicate the insulin-signaling pathway in the transduction of environmental factors into condition-dependent patterns of wing growth in insects.