RESUMO
In holometabolous insects, extensive reorganisation of tissues and cells occurs at the pupal stage. The remodelling of the external exoskeleton and internal organs that intervenes during metamorphosis has been traditionally studied in many insect species based on histological or ultrastructural methods. This study demonstrates the use of synchrotron X-ray phase-contrast micro-computed tomography as a powerful, non-destructive tool for in situ morphological observation of anatomical structures at the pupal stage in two Tenebrionid beetles, i.e. Tribolium castaneum and Tenebrio molitor, known as important pests, as well as emerging and promising models in experimental biology. Virtual sections and three-dimensional reconstructions were performed on both males and females at early, intermediate, and late pupal stage. The dataset allowed us to observe the remodelling of the gut and nervous system as well as the shaping of the female and male reproductive system at different pupal ages in both mealworm and red flour beetles. Moreover, we observed that the timing and duration pattern of organ development varied between the species analysed, likely related to the species-specific adaptations of the pre-imaginal stages to environmental conditions, which ultimately affect their life cycle. This research provides new knowledge on the morphological modifications that occur during the pupal stage of holometabolous insects and provides a baseline set of information on beetle metamorphosis that may support future research in forensics, physiology, and ecology as well as an image atlas for educational purposes.
Assuntos
Tenebrio , Tribolium , Animais , Masculino , Feminino , Tribolium/anatomia & histologia , Tribolium/fisiologia , Larva/fisiologia , Microtomografia por Raio-X , Metamorfose BiológicaRESUMO
The acquisition of novel traits is central to organismal evolution, yet the molecular mechanisms underlying this process are elusive. The beetle forewings (elytra) are evolutionarily modified to serve as a protective shield, providing a unique opportunity to study these mechanisms. In the past, the orthologs of genes within the wing gene network from Drosophila studies served as the starting point when studying the evolution of elytra (candidate genes). Although effective, candidate gene lists are finite and only explore genes conserved across species. To go beyond candidate genes, we used RNA sequencing and explored the wing transcriptomes of two Coleopteran species, the red flour beetle (Tribolium castaneum) and the Japanese stag beetle (Dorcus hopei). Our analysis revealed sets of genes enriched in Tribolium elytra (57 genes) and genes unique to the hindwings, which possess more "typical" insect wing morphologies (29 genes). Over a third of the hindwing-enriched genes were "candidate genes" whose functions were previously analyzed in Tribolium, demonstrating the robustness of our sequencing. Although the overlap was limited, transcriptomic comparison between the beetle species found a common set of genes, including key wing genes, enriched in either elytra or hindwings. Our RNA interference analysis for elytron-enriched genes in Tribolium uncovered novel genes with roles in forming various aspects of morphology that are unique to elytra, such as pigmentation, hardening, sensory development, and vein formation. Our analyses deepen our understanding of how gene network evolution facilitated the emergence of the elytron, a unique structure critical to the evolutionary success of beetles.
Assuntos
Besouros , Tribolium , Animais , Besouros/genética , Transcriptoma , Tribolium/genética , Tribolium/anatomia & histologia , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Asas de Animais , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismoRESUMO
In the postgenomics era, comparative genomics have advanced the understanding of evolutionary processes of neuropeptidergic signaling systems. The evolutionary origin of many neuropeptidergic signaling systems can be traced date back to early metazoan evolution based on the conserved sequences. Insect parathyroid hormone receptor (iPTHR) was previously described as an ortholog of vertebrate PTHR that has a well-known function in controlling bone remodeling. However, there was no sequence homologous to PTH sequence in insect genomes, leaving the iPTHR as an orphan receptor. Here, we identified the authentic ligand insect PTH (iPTH) for the iPTHR. The taxonomic distribution of iPTHR, which is lacking in Diptera and Lepidoptera, provided a lead for identifying the authentic ligand. We found that a previously described orphan ligand known as PXXXamide (where X is any amino acid) described in the cuttlefish Sepia officinalis has a similar taxonomic distribution pattern as iPTHR. Tests of this peptide, iPTH, in functional reporter assays confirmed the interaction of the ligand-receptor pair. Study of a model beetle, Tribolium castaneum, was used to investigate the function of the iPTH signaling system by RNA interference followed by RNA sequencing and phenotyping. The results suggested that the iPTH system is likely involved in the regulation of cuticle formation that culminates with a phenotype of defects in wing exoskeleton maturation at the time of adult eclosion. Moreover, RNAi of iPTHRs also led to significant reductions in egg numbers and hatching rates after parental RNAi.
Assuntos
Neuropeptídeos/metabolismo , Hormônio Paratireóideo/metabolismo , Receptores de Hormônios Paratireóideos/genética , Tribolium/anatomia & histologia , Animais , Evolução Molecular , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Fenótipo , Filogenia , Receptores de Hormônios Paratireóideos/metabolismo , Análise de Sequência de RNA , Tribolium/genética , Tribolium/metabolismo , Asas de Animais/anatomia & histologiaRESUMO
Male genitals are highly divergent in animals with internal fertilization. Most studies attempting to explain this diversity have focused on testing the major hypotheses of genital evolution (the lock-and-key, pleiotropy, and sexual selection hypotheses), and quantifying the form of selection targeting male genitals has played an important role in this endeavor. However, we currently know far less about selection targeting female genitals or how male and female genitals interact during mating. Here, we use formal selection analysis to show that genital size and shape is subject to strong multivariate stabilizing sexual selection in both sexes of the red flour beetle, Tribolium castaneum. Moreover, we show significant sexual selection on the covariance between the sexes for specific aspects of genital shape suggesting that male and female genitalia also interact to determine the successful transfer of a spermatophore during mating. Our work therefore highlights the important role that both male and female genital morphologies play in determining mating success and that these effects can occur independently, as well as through their interaction. Moreover, it cautions against the overly simplistic view that the sexual selection targeting genital morphology will always be directional in form and restricted primarily to males.
Assuntos
Evolução Biológica , Seleção Sexual , Tribolium/anatomia & histologia , Animais , Feminino , Genitália Feminina/anatomia & histologia , Genitália Masculina/anatomia & histologia , MasculinoRESUMO
Purpose: To reveal the effects of Ultraviolet-C (UV-C) on the elytra and hindwing morphology of Tribolium castaneum. Material and methods: Zero-day-old-pupae were irradiated with UV-C at a distance of 35 cm for 1, 2, 4, 8, 16, 32, or 64 min. Changes in wing morphologies were examined using light and scanning electron microscope. Results: UV-C radiation decreased the adult emergence rate and the insect body mass. Morphological changes of the elytra and hindwings in the adults were classified into nine grades. The treated insects had wrinkled and split elytra, and hindwings were not folded properly. Radiation altered the size of elytra, hindwings and wing shape. An analysis of the color intensity indicated that the irradiated beetles had darker elytra. The veins of hindwings became darker, while the membranous area had a lighter color than the control. UV-C radiation also affected the thickness of the elytra. Scanning electron microscopy revealed that UV-C caused deformity of elytra surface and decreased the number of hair sensilla. Conclusions: Results indicate that the elytra and hindwing morphology were altered by UV-C radiation. However, further analysis is required to evaluate the response of T. castaneum to UV-C radiation at the gene level.
Assuntos
Controle de Pragas/métodos , Pupa/anatomia & histologia , Pupa/efeitos da radiação , Tribolium/anatomia & histologia , Tribolium/efeitos da radiação , Raios Ultravioleta , Asas de Animais/efeitos da radiação , Animais , Tamanho Corporal/efeitos da radiação , Tamanho do Órgão/efeitos da radiação , Pigmentação/efeitos da radiação , Tribolium/crescimento & desenvolvimento , Asas de Animais/anatomia & histologia , Asas de Animais/metabolismoRESUMO
Body size in holometabolous insects is determined by the size at which the juvenile larva undergoes metamorphosis to the pupal stage. To undergo larva-pupa transition, larva must reach a critical developmental checkpoint, the threshold size (TS); however, the molecular mechanisms through which the TS cues this transition remain to be fully characterized. Here, we use the flour beetle Tribolium castaneum to characterize the molecular mechanisms underlying entry into metamorphosis. We found that T. castaneum reaches a TS at the beginning of the last larval instar, which is associated with the downregulation of TcKr-h1 and the upregulation of TcE93 and TcBr-C. Unexpectedly, we found that while there is an association between TS and TcE93 upregulation, it is the latter that constitutes the molecular trigger for metamorphosis initiation. In light of our results, we evaluate the interactions that control the larva-pupa transition and suggest alternative models.
Assuntos
Proteínas de Insetos/fisiologia , Metamorfose Biológica/genética , Tribolium/genética , Animais , Tamanho Corporal , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/anatomia & histologia , Larva/genética , Larva/crescimento & desenvolvimento , Tribolium/anatomia & histologia , Tribolium/crescimento & desenvolvimento , Regulação para CimaRESUMO
Despite the immense importance of the wing in the evolution and successful radiation of the insect lineages, the origin of this critical structure remains a hotly-debated mystery. Two possible tissues have been identified as an evolutionary origin of wings; the lateral expansion of the dorsal body wall (tergal edge) and structures related to an ancestral proximal leg segment (pleural tissues). Through studying wing-related tissues in the red flour beetle, Tribolium castaneum, we have previously presented evidence in support of a dual origin of insect wings, a third hypothesis proposing that wings evolved from a combination of both tergal and pleural tissues. One key finding came from the investigation of a Cephalothorax (Cx) mutant, in which the ectopic wing characteristic to this mutant was found to be formed from both tergal and pleural contributions. However, the degree of contribution of the two tissues to the wing remains elusive. Here, we took advantage of multiple Cx alleles available in Tribolium, and produced a variety of degrees and types of ectopic wing tissues in their prothoracic segments. Through detailed phenotypic scoring of the Cx phenotypes based on nine categories of mutant traits, along with comprehensive morphological analysis of the ectopic wing tissues, we found that (i) ectopic wing tissues can be formed at various locations in the prothorax, even internally, (ii) the lateral external ectopic wing tissues have tergal origin, while the internal and posterior external ectopic wing tissues appear to be of pleural origin, and (iii) the ectopic wing tissues of both tergal and pleural origin are capable of transforming into wing surface tissues. Collectively, these outcomes suggest that the evolutionary contribution of each tissue to a complete wing may be more complex than the simple binary view that is typically invoked by a dual origin model (i.e. the wing blade from the tergal contribution + musculature and articulation from the pleural contribution).
Assuntos
Evolução Biológica , Proteínas de Homeodomínio/genética , Proteínas de Insetos/genética , Tribolium/crescimento & desenvolvimento , Asas de Animais/crescimento & desenvolvimento , Animais , Proteínas de Homeodomínio/metabolismo , Proteínas de Insetos/metabolismo , Mutação , Tribolium/anatomia & histologia , Tribolium/genética , Asas de Animais/anatomia & histologiaRESUMO
Several recent studies have elucidated the molecular mechanisms that confer insecticide resistance on insect pests. However, little is known about multiple resistance in red flour beetle (Tribolium castaneum) at molecular level. The multiple resistance is characterized as resistance to different classes of insecticides that have different target sites, and is mediated by several enzymatic systems. In this study, we investigated the biochemical and molecular mechanisms involved in multiple resistance of T. castaneum to bifenthrin (pyrethroid [Pyr]) and pirimiphos-methyl (organophosphate [Org]). We used artificial selection, biochemical and in silico approaches including structural computational biology. After five generations of artificial selection in the presence of bifenthrin (F5Pyr) or pirimiphos-methyl (F5Org), we found high levels of multiple resistance. The hierarchical enzymatic cluster revealed a pool of esterases (E), lipases (LIPs) and laccase2 (LAC2) potentially contributing to the resistance in different ways throughout development, after one or more generations in the presence of insecticides. The enzyme-insecticide interaction network indicated that E2, E3, LIP3, and LAC2 are enzymes potentially required for multiple resistance phenotype. Kinetic analysis of esterases from F5Pyr and F5Org showed that pirimiphos-methyl and specially bifenthrin promote enzyme inhibition, indicating that esterases mediate resistance by sequestering bifenthrin and pirimiphos-methyl. Our computational data were in accordance with kinetic results, indicating that bifenthrin has higher affinity at the active site of esterase than pirimiphos-methyl. We also report the capability of these insecticides to modify the development in T. castaneum. Our study provide insights into the biochemical mechanisms employed by T. castaneum to acquire multiple resistance.
Assuntos
Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Proteínas de Insetos/metabolismo , Resistência a Inseticidas/efeitos dos fármacos , Compostos Organotiofosforados/toxicidade , Piretrinas/toxicidade , Tribolium/enzimologia , Animais , Eletroforese em Gel de Poliacrilamida , Esterases/classificação , Esterases/genética , Esterases/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/genética , Inseticidas/toxicidade , Isoenzimas/classificação , Isoenzimas/genética , Isoenzimas/metabolismo , Cinética , Lacase/classificação , Lacase/genética , Lacase/metabolismo , Lipase/classificação , Lipase/genética , Lipase/metabolismo , Modelos Moleculares , Filogenia , Domínios Proteicos , Pupa/anatomia & histologia , Pupa/efeitos dos fármacos , Pupa/enzimologia , Tribolium/anatomia & histologia , Tribolium/metabolismoRESUMO
Rensch's rule proposes a universal allometric scaling phenomenon across species where sexual size dimorphism (SSD) has evolved: in taxa with male-biased dimorphism, degree of SSD should increase with overall body size, and in taxa with female-biased dimorphism, degree of SSD should decrease with increasing average body size. Rensch's rule appears to hold widely across taxa where SSD is male-biased, but not consistently when SSD is female-biased. Furthermore, studies addressing this question within species are rare, so it remains unclear whether this rule applies at the intraspecific level. We assess body size and SSD within Tribolium castaneum (Herbst), a species where females are larger than males, using 21 populations derived from separate locations across the world, and maintained in isolated laboratory culture for at least 20 years. Body size, and hence SSD patterns, are highly susceptible to variations in temperature, diet quality and other environmental factors. Crucially, here we nullify interference of such confounds as all populations were maintained under identical conditions (similar densities, standard diet and exposed to identical temperature, relative humidity and photoperiod). We measured thirty beetles of each sex for all populations, and found body size variation across populations, and (as expected) female-biased SSD in all populations. We test whether Rensch's rule holds for our populations, but find isometry, i.e. no allometry for SSD. Our results thus show that Rensch's rule does not hold across populations within this species. Our intraspecific test matches previous interspecific studies showing that Rensch's rule fails in species with female-biased SSD.
Assuntos
Tribolium/anatomia & histologia , Animais , Tamanho Corporal , Feminino , Masculino , Caracteres SexuaisRESUMO
The adult insect brain is composed of neuropils present in most taxa. However, the relative size, shape, and developmental timing differ between species. This diversity of adult insect brain morphology has been extensively described while the genetic mechanisms of brain development are studied predominantly in Drosophila melanogaster. However, it has remained enigmatic what cellular and genetic mechanisms underlie the evolution of neuropil diversity or heterochronic development. In this perspective paper, we propose a novel approach to study these questions. We suggest using genome editing to mark homologous neural cells in the fly D. melanogaster, the beetle Tribolium castaneum, and the Mediterranean field cricket Gryllus bimaculatus to investigate developmental differences leading to brain diversification. One interesting aspect is the heterochrony observed in central complex development. Ancestrally, the central complex is formed during embryogenesis (as in Gryllus) but in Drosophila, it arises during late larval and metamorphic stages. In Tribolium, it forms partially during embryogenesis. Finally, we present tools for brain research in Tribolium including 3D reconstruction and immunohistochemistry data of first instar brains and the generation of transgenic brain imaging lines. Further, we characterize reporter lines labeling the mushroom bodies and reflecting the expression of the neuroblast marker gene Tc-asense, respectively.
Assuntos
Edição de Genes/métodos , Insetos/anatomia & histologia , Neurópilo/classificação , Animais , Animais Geneticamente Modificados , Evolução Biológica , Encéfalo/anatomia & histologia , Encéfalo/crescimento & desenvolvimento , Encéfalo/fisiologia , Insetos/classificação , Insetos/crescimento & desenvolvimento , Insetos/fisiologia , Larva/anatomia & histologia , Corpos Pedunculados/anatomia & histologia , Corpos Pedunculados/fisiologia , Neurópilo/citologia , Tribolium/anatomia & histologia , Tribolium/genética , Tribolium/crescimento & desenvolvimento , Tribolium/fisiologiaRESUMO
Morphological innovation is a fundamental process in evolution, yet its molecular basis is still elusive. Acquisition of elytra, highly modified beetle forewings, is an important innovation that has driven the successful radiation of beetles. Our RNAi screening for candidate genes has identified abrupt (ab) as a potential key player in elytron evolution. In this study, we performed a series of RNA interference (RNAi) experiments in both Tribolium and Drosophila to understand the contributions of ab to the evolution of beetle elytra. We found that (i) ab is essential for proper wing vein patterning both in Tribolium and Drosophila, (ii) ab has gained a novel function in determining the unique elytron shape in the beetle lineage, (iii) unlike Hippo and Insulin, other shape determining pathways, the shape determining function of ab is specific to the elytron and not required in the hindwing, (iv) ab has a previously undescribed role in the Notch signal-associated wing formation processes, which appears to be conserved between beetles and flies. These data suggest that ab has gained a new function during elytron evolution in beetles without compromising the conserved wing-related functions. Gaining a new function without losing evolutionarily conserved functions may be a key theme in the evolution of morphologically novel structures.
Assuntos
Evolução Biológica , Sequência Conservada , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/anatomia & histologia , Proteínas de Insetos/metabolismo , Proteínas Nucleares/metabolismo , Tribolium/anatomia & histologia , Asas de Animais/anatomia & histologia , Asas de Animais/crescimento & desenvolvimento , Animais , Padronização Corporal , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/crescimento & desenvolvimento , Genes de Insetos , Proteínas de Insetos/genética , Larva , Proteínas Nucleares/genética , Fenótipo , Interferência de RNA , Receptores Notch/metabolismo , Tribolium/genética , Tribolium/crescimento & desenvolvimentoRESUMO
The iBeetle-Base (http://ibeetle-base.uni-goettingen.de) makes available annotations of RNAi phenotypes, which were gathered in a large scale RNAi screen in the red flour beetle Tribolium castaneum (iBeetle screen). In addition, it provides access to sequence information and links for all Tribolium castaneum genes. The iBeetle-Base contains the annotations of phenotypes of several thousands of genes knocked down during embryonic and metamorphic epidermis and muscle development in addition to phenotypes linked to oogenesis and stink gland biology. The phenotypes are described according to the EQM (entity, quality, modifier) system using controlled vocabularies and the Tribolium morphological ontology (TrOn). Furthermore, images linked to the respective annotations are provided. The data are searchable either for specific phenotypes using a complex 'search for morphological defects' or a 'quick search' for gene names and IDs. The red flour beetle Tribolium castaneum has become an important model system for insect functional genetics and is a representative of the most species rich taxon, the Coleoptera, which comprise several devastating pests. It is used for studying insect typical development, the evolution of development and for research on metabolism and pest control. Besides Drosophila, Tribolium is the first insect model organism where large scale unbiased screens have been performed.
Assuntos
Bases de Dados Genéticas , Genes de Insetos , Interferência de RNA , Tribolium/genética , Animais , Feminino , Internet , Fenótipo , Tribolium/anatomia & histologia , Tribolium/embriologia , Interface Usuário-ComputadorRESUMO
In a morphological ontology the expert's knowledge is represented in terms, which describe morphological structures and how these structures relate to each other. With the assistance of ontologies this expert knowledge is made processable by machines, through a formal and standardized representation of terms and their relations to each other. The red flour beetle Tribolium castaneum, a representative of the most species rich animal taxon on earth (the Coleoptera), is an emerging model organism for development, evolution, physiology, and pest control. In order to foster Tribolium research, we have initiated the Tribolium Ontology (TrOn), which describes the morphology of the red flour beetle. The content of this ontology comprises so far most external morphological structures as well as some internal ones. All modeled structures are consistently annotated for the developmental stages larva, pupa and adult. In TrOn all terms are grouped into three categories: Generic terms represent morphological structures, which are independent of a developmental stage. In contrast, downstream of such terms are concrete terms which stand for a dissectible structure of a beetle at a specific life stage. Finally, there are mixed terms describing structures that are only found at one developmental stage. These terms combine the characteristics of generic and concrete terms with features of both. These annotation principles take into account the changing morphology of the beetle during development and provide generic terms to be used in applications or for cross linking with other ontologies and data resources. We use the ontology for implementing an intuitive search function at the electronic iBeetle-Base, which stores morphological defects found in a genome wide RNA interference (RNAi) screen. The ontology is available for download at http://ibeetle-base.uni-goettingen.de.
Assuntos
Ferramenta de Busca/métodos , Tribolium/classificação , Animais , Internet , Tribolium/anatomia & histologia , Tribolium/genéticaRESUMO
BACKGROUND: RNA interference (RNAi) is a powerful tool to study gene function in organisms that are not amenable to classical forward genetics. Hence, together with the ease of comprehensively identifying genes by new generation sequencing, RNAi is expanding the scope of animal species and questions that can be addressed in terms of gene function. In the case of genetic mutants, the genetic background of the strains used is known to influence the phenotype while this has not been described for RNAi experiments. RESULTS: Here we show in the red flour beetle Tribolium castaneum that RNAi against Tc-importin α1 leads to different phenotypes depending on the injected strain. We rule out off target effects and show that sequence divergence does not account for this difference. By quantitatively comparing phenotypes elicited by RNAi knockdown of four different genes we show that there is no general difference in RNAi sensitivity between these strains. Finally, we show that in case of Tc-importin α1 the difference depends on the maternal genotype. CONCLUSIONS: These results show that in RNAi experiments strain specific differences have to be considered and that a proper documentation of the injected strain is required. This is especially important for the increasing number of emerging model organisms that are being functionally investigated using RNAi. In addition, our work shows that RNAi is suitable to systematically identify the differences in the gene regulatory networks present in populations of the same species, which will allow novel insights into the evolution of animal diversity.
Assuntos
Fenótipo , Interferência de RNA , Tribolium/genética , Sequência de Aminoácidos , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Técnicas de Silenciamento de Genes , Injeções , Proteínas de Insetos/química , Proteínas de Insetos/deficiência , Proteínas de Insetos/genética , Masculino , Dados de Sequência Molecular , Mães , Especificidade da Espécie , Tribolium/anatomia & histologia , Tribolium/embriologia , alfa Carioferinas/química , alfa Carioferinas/deficiência , alfa Carioferinas/genéticaRESUMO
BACKGROUND: The ATP-binding cassette (ABC) transporters belong to a large superfamily of proteins that have important physiological functions in all living organisms. Most are integral membrane proteins that transport a broad spectrum of substrates across lipid membranes. In insects, ABC transporters are of special interest because of their role in insecticide resistance. RESULTS: We have identified 73 ABC transporter genes in the genome of T. castaneum, which group into eight subfamilies (ABCA-H). This coleopteran ABC family is significantly larger than those reported for insects in other taxonomic groups. Phylogenetic analysis revealed that this increase is due to gene expansion within a single clade of subfamily ABCC. We performed an RNA interference (RNAi) screen to study the function of ABC transporters during development. In ten cases, injection of double-stranded RNA (dsRNA) into larvae caused developmental phenotypes, which included growth arrest and localized melanization, eye pigmentation defects, abnormal cuticle formation, egg-laying and egg-hatching defects, and mortality due to abortive molting and desiccation. Some of the ABC transporters we studied in closer detail to examine their role in lipid, ecdysteroid and eye pigment transport. CONCLUSIONS: The results from our study provide new insights into the physiological function of ABC transporters in T. castaneum, and may help to establish new target sites for insect control.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Genômica , Proteínas de Insetos/genética , Tribolium/genética , Transportadores de Cassetes de Ligação de ATP/deficiência , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Olho/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Proteínas de Insetos/deficiência , Proteínas de Insetos/metabolismo , Larva/anatomia & histologia , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Metabolismo dos Lipídeos/genética , Masculino , Fenótipo , Pigmentação/genética , Interferência de RNA , Tribolium/anatomia & histologia , Tribolium/crescimento & desenvolvimento , Tribolium/metabolismoRESUMO
The morphological features of the third instar larva of the most important insect model, Drosophila melanogaster, are documented for the first time using a broad spectrum of modern morphological techniques. External structures of the body wall, the cephaloskeleton, and the musculature are described and illustrated. Additional information about other internal organs is provided. The systematic implications of the findings are discussed briefly. Internal apomorphic features of Brachycera and Cyclorrhapha are confirmed for Drosophila. Despite the intensive investigations of the phylogeny of the megadiverse Diptera, evolutionary reconstructions are still impeded by the scarcity of anatomical data for brachyceran larvae. The available morphological information for the life stages of three insect model organisms -D. melanogaster (Diptera, Drosophilidae), Manduca sexta (Lepidoptera, Sphingidae) and Tribolium castaneum (Coleoptera, Tenebrionidae) - is addressed briefly. The usefulness of a combination of traditional and innovative techniques for an optimized acquisition of anatomical data for different life stages is highlighted.
Assuntos
Drosophila melanogaster/anatomia & histologia , Animais , Drosophila melanogaster/ultraestrutura , Larva/anatomia & histologia , Larva/ultraestrutura , Manduca/anatomia & histologia , Manduca/ultraestrutura , Microscopia Eletrônica de Varredura , Modelos Animais , Filogenia , Tribolium/anatomia & histologia , Tribolium/ultraestruturaRESUMO
Cephalization and seamless fusion of the anterior body segments during development obscure the segmental boundaries of the insect head. Most of the visible seams are thought to reflect cuticular infolding for structural reinforcement rather than a merger of cuticular plate borders. Incomplete fusions and other modifications of the adult head found in eight Tribolium mutations indicate that the frontal and gular sutures likely are true sutures that mark borders between adjacent cuticular plates, and suggest that the anterior facial shelf is a composite of three independent cuticular surfaces: ocular, antennal, and clypeo-labral. Additionally, midline splits of the clypeo-labrum and gula, and membranous lesions on the lateral head capsule reveal probable borders of adjacent cuticular plates where visible sutures are normally absent. The anterior lateral lesions seen in the Lucifer mutation mark a border between ocular and antennal plates and appear to identify part of the postfrontal sutures. While revealing or clarifying possible intersegmental borders between ocular, antennal, and clypeo-labral plates, the various modified or unfused surfaces of the head neither reveal an additional acronal plate nor support the view that the clypeo-labrum is segmentally associated with ocular cuticle.
Assuntos
Tribolium/anatomia & histologia , Tribolium/genética , Animais , Evolução Biológica , Padronização Corporal , Cabeça/anatomia & histologia , Cabeça/crescimento & desenvolvimento , Mutação , Tribolium/crescimento & desenvolvimentoRESUMO
Red flour beetle (T. castaneum) is a major pest of stored grains and is known for its adaptability to all classes of insecticides. The present study was carried out to determine the insecticidal potential of egg white proteins to manage beetle population. Protein samples obtained through salt fractionation were lyophilized and were used separately and simultaneously in different concentrations by adding them to wheat flour and milk powder. The results indicated that the mortality rate of the adult beetles was dependent on the type of treatment, concentration of protein samples and duration of feeding. In multiple-choice feeding trials beetles showed their movement towards the control section as the concentration of treatment increases. Marked abnormalities were observed in appearance and dimensions of the testes which indicated that the egg white proteins caused considerable effect on the process of spermatogenesis and sperm functions. SEM study revealed the formation of deep wrinkles and folds on the testicular surface of the testes of beetles fed on treated diets, points towards the depletion of internal cellular material. The results suggest that egg white protein affects the survival and cause subsequent derangements in the testis of red flour beetle.
Assuntos
Proteínas do Ovo/farmacologia , Proteínas do Ovo/toxicidade , Testículo/anormalidades , Testículo/efeitos dos fármacos , Tribolium/anatomia & histologia , Tribolium/efeitos dos fármacos , Animais , Galinhas , Comportamento Alimentar , Masculino , Testículo/ultraestruturaRESUMO
We report that the abdominal epidermis and associated tissues are the predominant sources of male-produced pheromones in the red flour beetle, Tribolium castaneum and, for the first time, describe the stereoisomeric composition of the natural blend of isomers of the aggregation pheromone 4,8-dimethyldecanal (DMD) in this important pest species. Quantitative analyses via gas chromatography-mass spectrometry showed that the average amount of DMD released daily by single feeding males of T. castaneum was 878 ± 72 ng (SE). Analysis of different body parts identified the abdominal epidermis as the major source of aggregation pheromone; the thorax was a minor source, while no DMD was detectable in the head. No internal organs or obvious male-specific glands were associated with pheromone deposition. Complete separation of all four stereoisomers of DMD was achieved following oxidation to the corresponding acid, derivatization with (1R, 2R)- and (1S, 2S)-2-(anthracene-2,3-dicarboximido)cyclohexanol to diastereomeric esters, and their separation on reversed-phase high-performance liquid chromatography at -54°C. Analysis of the hexane eluate from Porapak-Q-collected volatiles from feeding males revealed the presence of all four isomers (4R,8R)/(4R,8S)/(4S,8R)/(4S,8S) at a ratio of approximately 4:4:1:1. A walking orientation bioassay in a wind tunnel with various blends of the four synthetic isomers further indicated that the attractive potency of the reconstituted natural blend of 4:4:1:1 was equivalent to that of the natural pheromone and greater than that of the 1:1 blend of (4R,8R)/(4R,8S) used in commercial lures.
Assuntos
Feromônios/química , Tribolium/anatomia & histologia , Tribolium/química , Aldeídos/química , Animais , Cromatografia Líquida de Alta Pressão , Epiderme/metabolismo , Masculino , Estereoisomerismo , Tribolium/metabolismoRESUMO
Sexual size dimorphism (SSD) is a conspicuous yet poorly understood pattern across many organisms. Although artificial selection is an important tool for studying the evolution of SSD, previous studies have applied selection to only a single sex or to both sexes in the same direction. In nature, however, SSD likely arises through sex-specific selection on body size. Here, we use Tribolium castaneum flour beetles to investigate the evolution of SSD by subjecting males and females to sexually antagonistic selection on body size (sexes selected in opposite directions). Additionally, we examined correlated responses to body size selection in larval growth rates and development time. After seven generations, SSD remained unchanged in all selected lines; this observed lack of response to short-term selection may be attributed to evolutionary constraints arising from between-sex body size correlations. Developmental traits showed complex correlated responses under different selection treatments. These results suggest that sex-specific larval development patterns may facilitate the evolution of SSD.