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1.
Gene ; 850: 146917, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36174905

RESUMO

Among bioluminescent beetles of the Elateroidea superfamily, Phengodidae is the third largest family, with 244 bioluminescent species distributed only in the Americas, but is still the least studied from the phylogenetic and evolutionary points of view. The railroad worm Phrixothrix hirtus is an essential biological model and symbolic species due to its bicolor bioluminescence, being the only organism that produces true red light among bioluminescent terrestrial species. Here, we performed partial genome assembly of P. hirtus, combining short and long reads generated with Illumina sequencing, providing the first source of genomic information and a framework for comparative analyses of the bioluminescent system in Elateroidea. This is the largest genome described in the Elateroidea superfamily, with an estimated size of ∼3.4 Gb, displaying 32 % GC content, and 67 % transposable elements. Comparative genomic analyses showed a positive selection of genes and gene family expansion events of growth and morphogenesis gene products, which could be associated with the atypical anatomical development and morphogenesis found in paedomorphic females and underdeveloped males. We also observed gene family expansion among distinct odorant-binding receptors, which could be associated with the pheromone communication system typical of these beetles, and retrotransposable elements. Common genes putatively regulating bioluminescence production and control, including two luciferase genes corresponding to lateral lanterns green-emitting and head lanterns red-emitting luciferases with 7 exons and 6 introns, and genes potentially involved in luciferin biosynthesis were found, indicating that there are no clear differences about the presence or absence of gene families associated with bioluminescence in Elateroidea.


Assuntos
Besouros , Ferrovias , Animais , Feminino , Filogenia , Elementos de DNA Transponíveis , Odorantes , Besouros/genética , Besouros/metabolismo , Luciferases/metabolismo , Morfogênese , Feromônios
2.
Biomolecules ; 12(11)2022 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-36358934

RESUMO

Hypothermic (cold) preservation is a limiting factor for successful cell and tissue transplantation where cell swelling (edema) usually develops, impairing cell function. University of Wisconsin (UW) solution, a standard cold preservation solution, contains effective components to suppress hypothermia-induced cell swelling. Antifreeze proteins (AFPs) found in many cold-adapted organisms can prevent cold injury of the organisms. Here, the effects of a beetle AFP from Dendroides canadensis (DAFP-1) on pancreatic ß-cells preservation were first investigated. As low as 500 µg/mL, DAFP-1 significantly minimized INS-1 cell swelling and subsequent cell death during 4 °C preservation in UW solution for up to three days. However, such significant cytoprotection was not observed by an AFP from Tenebrio molitor (TmAFP), a structural homologue to DAFP-1 but lacking arginine, at the same levels. The cytoprotective effect of DAFP-1 was further validated with the primary ß-cells in the isolated rat pancreatic islets in UW solution. The submilligram level supplement of DAFP-1 to UW solution significantly increased the islet mass recovery after three days of cold preservation followed by rewarming. The protective effects of DAFP-1 in UW solution were discussed at a molecular level. The results indicate the potential of DAFP-1 to enhance cell survival during extended cold preservation.


Assuntos
Besouros , Animais , Ratos , Besouros/química , Besouros/metabolismo , Sobrevivência Celular , alfa-Fetoproteínas/farmacologia , Proteínas Anticongelantes/química , Glutationa/farmacologia , Insulina/farmacologia , Edema
3.
Molecules ; 27(20)2022 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-36296556

RESUMO

Edible insects are a potential alternative food source of high feed conversion efficiency and protein content. Zophobas atratus is an edible insect that adapts to different diets, enabling sustainable rearing by adding value to by-products and agro-industrial residues. This study aimed to evaluate the performance and nutritional characterization of Zophobas atratus larvae fed with different proportions of grape residue. Physicochemical analysis of the diets and larvae (AOAC procedures), fatty acid profile (chromatographic techniques), metals and non-metals (inductively coupled plasma optical emission spectrometry), larval mass gain, feed conversion efficiency, and mortality rate were assessed. The replacement of 25% of the conventional diet with grape residue increased lipid, ash, and fiber contents and reduced protein, carbohydrates, and energy. It promoted greater mass gain, lower mortality rate, and reduced larval growth time by 51%. Among the replacements, 25% resulted in the second-highest content of calcium, sodium, magnesium, and zinc, and the lowest content of potassium and phosphorus in the larvae. The 100% replacement resulted in the highest amounts of C18:2n6 (27.8%), C18:3n3 (2.2%), and PUFA (30.0%). Replacing 25% of the conventional diet with grape residue is equivalent to the conventional diet in many aspects and improves several larvae performance indices and nutritional values.


Assuntos
Cálcio , Besouros , Animais , Larva , Cálcio/metabolismo , Magnésio/metabolismo , Besouros/metabolismo , Ração Animal/análise , Ácidos Graxos/metabolismo , Fósforo/metabolismo , Carboidratos , Zinco/metabolismo , Sódio/metabolismo , Potássio/metabolismo
4.
Pestic Biochem Physiol ; 187: 105180, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36127042

RESUMO

Bark beetles rely on detoxifying enzymes to resist the defensive terpenoids of the host tree. Insect cytochrome P450 (CYPs) plays a key role in the detoxification of pesticides and plant allelochemicals. CYP6 family is unique to Insecta, and its biochemical function is basically related to the metabolism of exogenous substances. In this study, we sequenced and characterized the full-length cDNAs of two CYP6 genes from Chinese white pine beetle, Dendroctonus armandi. Spatiotemporal expression profiling revealed that the expression of CYP6CR2 and CYP6DE5 was higher in larval and adult stages of D. armandi than that in other developmental stages, and that two genes predominantly expressed in brain, midgut, fat body, Malpighian tubules or hemolymph. The expression of CYP6CR2 and CYP6DE5 was significantly induced after feeding on the phloem of Pinus armandii and exposure to six stimuli [(±)- α -pinene, (-)-α-pinene, (-)-ß-pinene, (+)-3-carene, (±)-limonene and turpentine]. Importantly, silencing CYP6CR2 and CYP6DE5 separately could increase the sensitivity, led to a significant reduction of the activity of P450, resulting a significant increase in adult mortality after treatment with terpenoids. The comprehensive results of this study showed that in the process of host selection and colonization, the functions of CYPs were mainly to hydrolyze the chemical defense of the host and degrade odor molecules. These findings may help to develop new treatments to control this important pest.


Assuntos
Besouros , Praguicidas , Pinus , Animais , Monoterpenos Bicíclicos , China , Besouros/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Família 6 do Citocromo P450/metabolismo , Limoneno , Monoterpenos/metabolismo , Monoterpenos/farmacologia , Praguicidas/metabolismo , Feromônios/metabolismo , Terpenos , Terebintina/metabolismo
5.
J Proteome Res ; 21(10): 2247-2260, 2022 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-36107737

RESUMO

Neuropeptides are signaling molecules that regulate almost all physiological processes in animals. Around 50 different genes for neuropeptides have been described in insects. In Coleoptera, which is the largest insect order based on numbers of described species, knowledge about neuropeptides and protein hormones is still limited to a few species. Here, we analyze the neuropeptidomes of two closely related tenebrionid beetles: Tenebrio molitor and Zophobas atratus─both of which are model species in physiological and pharmacological research. We combined transcriptomic and mass spectrometry analyses of the central nervous system to identify neuropeptides and neuropeptide-like and protein hormones. Several precursors were identified in T. molitor and Z. atratus, of which 50 and 40, respectively, were confirmed by mass spectrometry. This study provides the basis for further functional studies of neuropeptides as well as for the design of environmentally friendly and species-specific peptidomimetics to be used as biopesticides. Furthermore, since T. molitor has become accepted by the European Food Safety Authority as a novel food, a deeper knowledge of the neuropeptidome of this species will prove useful for optimizing production programs at an industrial scale.


Assuntos
Besouros , Neuropeptídeos , Peptidomiméticos , Tenebrio , Animais , Agentes de Controle Biológico/metabolismo , Besouros/metabolismo , Hormônios , Larva/metabolismo , Neuropeptídeos/metabolismo , Peptidomiméticos/metabolismo , Tenebrio/genética , Tenebrio/metabolismo
6.
Chem Biol Interact ; 367: 110166, 2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36087814

RESUMO

Silver nanoparticles (AgNPs) have long been materials of great interest in numerous fields; however, there is escalating alarm over their toxicity to public health since exposure to these particles is inevitable. This study sheds light on the deleterious impacts of AgNPs on the midgut tissues of beetles (Blaps polychresta) collected from Egypt as a biological model. The investigations were conducted on the beetles administered with a sublethal dose of AgNPs (0.03 mg/g body weight) after 30 days. Oxidative stress parameters and antioxidant enzyme activities were assessed, which exposed critical disruption in the antioxidant defense system of treated beetles. Remarkably, metallothionein (MT) gene expression was significantly increased, while reduced glutathione (GSH) level was notably decreased in midgut tissues subjected to AgNPs. These findings manifestly imply the presence of overproduction in terms of reactive oxygen species (ROS) inside the cells. Additionally, DNA impairment and apoptosis of midgut cells were appraised employing comet and flow cytometry analyses, respectively. The comet results revealed a significant increase in comet cells for the AgNPs treated beetles compared with the control group. Furthermore, the apoptosis results demonstrated a substantial diminution in viable cells with significant growth in apoptotic cells in midgut cells exposed to AgNPs, manifesting their striking correlation with comet and biochemical findings. Noticeably, the histopathological and ultrastructural inspections revealed substantial aberrations in the midgut tissues in the AgNPs treated group, substantiating the previous results. As far as we know, no research has been found that surveyed how the AgNPs at low doses affect the midgut tissues of beetles. Overall, these findings evince the aberrant influences of AgNPs on living organisms.


Assuntos
Besouros , Nanopartículas Metálicas , Animais , Antioxidantes/metabolismo , Besouros/metabolismo , Glutationa/metabolismo , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Metalotioneína/genética , Metalotioneína/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Prata/química , Prata/toxicidade
7.
J Insect Physiol ; 142: 104429, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35964679

RESUMO

Eclosion hormone (EH) is a neurohormone that plays a key role in the regulation of insect pre-ecdysis behavior at the end of each molt. Previous research has reported more than one EH gene was found in certain insects, with their functions and mechanisms still unclear. Here, aside from the classical EH gene orthologous group, we characterized another novel orthologous cluster of eclosion hormone-like (EHL) genes in Arthropoda and investigated the roles of EHL during development in Tribolium castaneum. T. castaneum EHL (TcEHL) shows high expression levels during pupal - adult development, which also positively responded to 20-hydroxyecdysone (20E) treatment as well as RNA interference (RNAi) of ECR (20E nuclear receptor). Knockdown of TcEHL prevented the tanning of the adult cuticle and caused lethal phenotypes. Further analysis indicated that knockdown of TcEHL could upregulate expression levels of the classical TcEH, and downregulate the ecdysis behavior cascade genes, as well as tanning pathway enzymes. This suggests a critical role for TcEHL in adult eclosion and cuticle tanning. In addition, our data indicated that TcEHL is responsible for the female reproduction process. Taken together, these results suggest that TcEHL has specific roles in adult cuticle tanning during the post-eclosion process and female reproduction. They also suggest that EHL gene is the ancestral copy for the EH family and it is functionally shuffled by synfunctionalization.


Assuntos
Besouros , Hormônios de Inseto , Tribolium , Animais , Besouros/genética , Besouros/metabolismo , Ecdisterona/metabolismo , Feminino , Hormônios de Inseto/metabolismo , Interferência de RNA
8.
Contrast Media Mol Imaging ; 2022: 1892384, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35909589

RESUMO

Lung adenocarcinoma (LUAD) is one of the major causes of cancer death in the world. Studies show that the effective anticancer component in blister beetles is cantharidin, which can improve chemotherapy efficacy, median survival, and prognosis of LUAD. However, the antitumor mechanism of blister beetles has not been fully clarified. This study aimed to identify the key targets of the treatment of LUAD by blister beetles based on the principle of network pharmacology. An integrated approach including network pharmacology and a molecular docking technique was conducted, which mainly comprises target prediction, weighted gene correlation network analysis (WGCNA) analysis, network construction, gene ontology, and pathway enrichment analysis. 35 key targets were obtained and significantly associated with response to external stimuli, collagen binding, cyclin binding, organic acid binding, pyruvate metabolism, glycolysis, and amino acid biosynthesis pathways. Both LASSO regression and the RF model had a high predictive ability, and 9 candidate genes were screened, among which BIRC5 and PLK1 were the key targets for the treatment of LUAD by using blister beetles and showed significant survival significance. Cantharidin exerts its antitumor effects through 8 targets in 32 pathways, while BIRC5 and PLK1 have obvious survival significance.


Assuntos
Adenocarcinoma de Pulmão , Besouros , Neoplasias Pulmonares , Adenocarcinoma de Pulmão/tratamento farmacológico , Adenocarcinoma de Pulmão/genética , Animais , Cantaridina/química , Cantaridina/farmacologia , Cantaridina/uso terapêutico , Besouros/genética , Besouros/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Simulação de Acoplamento Molecular , Farmacologia em Rede
9.
Sci Rep ; 12(1): 14294, 2022 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-35995937

RESUMO

Biocontrol providing parasitoids can orientate according to volatile organic compounds (VOCs) of their host's plants, the emission of which is potentially dependent on the availability of soil nitrogen (N). This paper aimed at finding the optimal N fertilization rate for oilseed rape (Brassica napus L.) to favor parasitism of pollen beetles (Brassicogethes aeneus Fab. syn. Meligethes aeneus Fab.) in a controlled environment. Pollen beetles preferred to oviposit into buds of plants growing under higher N fertilization, whereas their parasitoids favored moderate N fertilization. As a part of induced defense, the proportion of volatile products of glucosinolate pathway in the total oilseed rape VOC emission blend was increased. Our results suggest that the natural biological control of pollen beetle herbivory is best supported by moderate N fertilization rates.


Assuntos
Brassica napus , Besouros , Animais , Besouros/metabolismo , Fertilização , Nitrogênio/metabolismo , Pólen
10.
Sci Rep ; 12(1): 14815, 2022 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-36045277

RESUMO

Luciferin biosynthetic origin and alternative biological functions during the evolution of beetles remain unknown. We have set up a bioluminescent sensing method for luciferin synthesis from cysteine and benzoquinone using E. coli and Pichia pastoris expressing the bright Amydetes vivianii firefly and P. termitilluminans click beetle luciferases. In the presence of D-cysteine and benzoquinone, intense bioluminescence is quickly produced, indicating the expected formation of D-luciferin. Starting with L-cysteine and benzoquinone, the bioluminescence is weaker and delayed, indicating that bacteria produce L-luciferin, and then racemize it to D-luciferin in the presence of endogenous esterases, CoA and luciferase. In bacteria the p-benzoquinone toxicity (IC50 ~ 25 µM) is considerably reduced in the presence of cysteine, maintaining cell viability at 3.6 mM p-benzoquinone concomitantly with the formation of luciferin. Transcriptional analysis showed the presence of gene products involved with the sclerotization/tanning in the photogenic tissues, suggesting a possible link between these pathways and bioluminescence. The lack of two enzymes involved with the last steps of these pathways, indicate the possible accumulation of toxic quinone intermediates in the lanterns. These results and the abundance of cysteine producing enzymes suggest that luciferin first appeared as a detoxification byproduct of cysteine reaction with accumulated toxic quinone intermediates during the evolution of sclerotization/tanning in Coleoptera.


Assuntos
Besouros , Luciferina de Vaga-Lumes , Luciferases de Vaga-Lume , Quinonas , Animais , Besouros/metabolismo , Cisteína/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Vaga-Lumes/genética , Luciferina de Vaga-Lumes/metabolismo , Luciferases/genética , Luciferases/metabolismo , Luciferases de Vaga-Lume/metabolismo , Luciferinas , Medições Luminescentes , Quinonas/metabolismo , Saccharomycetales/metabolismo
11.
Proc Natl Acad Sci U S A ; 119(33): e2205564119, 2022 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35943983

RESUMO

Male-female coevolution has taken different paths among closely related species, but our understanding of the factors that govern its direction is limited. While it is clear that ecological factors, life history, and the economics of reproduction are connected, the divergent links are often obscure. We propose that a complete understanding requires the conceptual integration of metabolic phenotypes. Metabolic rate, a nexus of life history evolution, is constrained by ecological factors and may exert important direct and indirect effects on the evolution of sexual dimorphism. We performed standardized experiments in 12 seed beetle species to gain a rich set of sex-specific measures of metabolic phenotypes, life history traits, and the economics of mating and analyzed our multivariate data using phylogenetic comparative methods. Resting metabolic rate (RMR) showed extensive evolution and evolved more rapidly in males than in females. The evolution of RMR was tightly coupled with a suite of life history traits, describing a pace-of-life syndrome (POLS), with indirect effects on the economics of mating. As predicted, high resource competition was associated with a low RMR and a slow POLS. The cost of mating showed sexually antagonistic coevolution, a hallmark of sexual conflict. The sex-specific costs and benefits of mating were predictably related to ecology, primarily through the evolution of male ejaculate size. Overall, our results support the tenet that resource competition affects metabolic processes that, in turn, have predictable effects on both life history evolution and reproduction, such that ecology shows both direct and indirect effects on male-female coevolution.


Assuntos
Metabolismo Basal , Evolução Biológica , Besouros , Comportamento Sexual Animal , Animais , Besouros/metabolismo , Feminino , Masculino , Filogenia , Reprodução
12.
Proc Natl Acad Sci U S A ; 119(30): e2201089119, 2022 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-35858446

RESUMO

Many insects enter a state of dormancy (diapause) during winter in which they lower their metabolism to save energy. Metabolic suppression is a hallmark of diapause, yet we know little about the mechanisms underpinning metabolic suppression in winter or how it is reversed in the spring. Here, we show that metabolic suppression in dormant Colorado potato beetles results from the breakdown of flight muscle mitochondria via mitophagy. Diapausing Colorado potato beetles suppress their metabolism by 90%, and this lowered metabolic rate coincides with a similar reduction in flight muscle mitochondrial function and density. During early diapause, beetles increase the expression of mitophagy-related transcripts (Parkin and ATG5) in their flight muscle coincident with an increase in mitophagy-related structures in the flight muscle. Knocking down Parkin expression with RNA interference in diapausing beetles prevented some mitochondrial breakdown and partially restored the whole animal metabolic rate, suggesting that metabolic suppression in diapausing beetles is driven by mitophagy. In other animals and in models of disease, such large-scale mitochondrial degradation is irreversible. However, we show that as diapause ends, beetles reverse mitophagy and increase the expression of PGC1α and NRF1 to replenish flight muscle mitochondrial pools. This mitochondrial biogenesis is activated in anticipation of diapause termination and in the absence of external stimuli. Our study provides a mechanistic link between mitochondrial degradation in insect tissues over the winter and whole-animal metabolic suppression.


Assuntos
Besouros , Diapausa de Inseto , Mitofagia , Animais , Proteína 5 Relacionada à Autofagia/genética , Proteína 5 Relacionada à Autofagia/metabolismo , Besouros/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias Musculares/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo
13.
Phys Chem Chem Phys ; 24(31): 18505-18510, 2022 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-35703330

RESUMO

Fireflies, click beetles, and railroad worms glow in the dark. The color varies from green to red among the insects and is associated with an electronically excited oxyluciferin formed catalytically by the luciferase enzyme. The actual color tuning mechanism has been, and still is, up for much debate. One complication is that oxyluciferin can occur in different charge states and isomeric forms. We present here emission spectra of oxyluciferin monoanions in vacuo at both room temperature and at 100 K recorded with a newly developed and unique mass-spectroscopy setup specially designed for gas-phase ion fluorescence spectroscopy. Ions are limited to the phenolate-keto and phenolate-enol forms that account for natural bioluminescence. At 100 K, fluorescence band maxima are at 599 ± 2 nm and 563 ± 2 nm for the keto and enol forms, respectively, and at 300 K about 5 nm further to the red. The bare-ion spectra, free from solvent effects, serve as important references as they reveal whether a protein microenvironment redshifts or blueshifts the emission, and they serve as important benchmarks for nontrivial excited-state calculations.


Assuntos
Besouros , Vaga-Lumes , Animais , Besouros/química , Besouros/metabolismo , Vaga-Lumes/química , Indóis/química , Luciferases/metabolismo , Pirazinas/química , Espectrometria de Fluorescência
14.
BMC Genomics ; 23(1): 461, 2022 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-35733103

RESUMO

BACKGROUND: Insect olfactory proteins can transmit chemical signals in the environment that serve as the basis for foraging, mate searching, predator avoidance and oviposition selection. Semanotus bifasciatus is an important destructive borer pest, but its olfactory mechanism is not clear. We identified the chemosensory genes of S. bifasciatus in China, then we conducted a phylogenetic analysis of the olfactory genes of S. bifasciatus and other species. And the expression profiles of odorant binding proteins (OBPs) genes in different tissues and different genders of S. bifasciatus were determined by quantitative real-time PCR for the first time. RESULTS: A total of 32 OBPs, 8 chemosensory proteins (CSPs), 71 odorant receptors (ORs), 34 gustatory receptors (GRs), 18 ionotropic receptors (IRs), and 3 sensory neuron membrane proteins (SNMPs) were identified. In the tissue expression analysis of OBP genes, 7 OBPs were higher expressed in antennae, among them, SbifOBP2, SbifOBP3, SbifOBP6, SbifOBP7 and SbifOBP20 were female-biased expression, while SbifOBP1 was male-biased expression and SbifOBP22 was no-biased expression in antennae. In addition, the expressed levels of SbifOBP4, SbifOBP12, SbifOBP15, SbifOBP27 and SbifOBP29 were very poor in the antennae, and SbifOBP4 and SbifOBP29 was abundant in the head or legs, and both of them were male-biased expression. While SbifOBP15 was highly expressed only at the end of the abdomen with its expression level in females three times than males. Other OBPs were expressed not only in antennae but also in various tissues. CONCLUSION: We identified 166 olfactory genes from S. bifasciatus, and classified these genes into groups and predicted their functions by phylogenetic analysis. The majority of OBPs were antenna-biased expressed, which are involved in odor recognition, sex pheromone detection, and/or host plant volatile detection. However, also some OBPs were detected biased expression in the head, legs or end of the abdomen, indicating that they may function in the different physiological processes in S. bifasciatus.


Assuntos
Besouros , Receptores Odorantes , Animais , Antenas de Artrópodes/metabolismo , Besouros/genética , Besouros/metabolismo , Feminino , Perfilação da Expressão Gênica , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Masculino , Odorantes , Filogenia , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Transcriptoma
15.
J Insect Physiol ; 140: 104405, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35679992

RESUMO

As the downstream effector of the target of rapamycin complex 1 (TORC1) signaling pathway, the ribosomal protein S6 kinase (S6K) is an important regulator of insect reproduction, however, the underlying mechanism remains obscure. In this study, a S6K gene, named TcS6K1, was isolated from the red flour beetle, Tribolium castaneum. Analysis of temporal and spatial expression patterns revealed that TcS6K1 is expressed at the highest level in the one-day-old first instar larvae and head of 7-day-old females, respectively. RNAi-mediated knockdown of TcS6K1 in either female or male adults decreased the number of eggs laid, with a concomitant reduction of mRNA levelsof vitellogenin genes, TcVg1 and TcVg2, two male accessory gland secretory proteins, as well as the juvenile hormone (JH) biosynthesis-related gene, farnesol dehydrogenase (TcFDH). While the mRNA and protein levels of the transcription factor forkhead box O (TcFOXO) were not affected, suppression of TcS6K1 expression promoted TcFOXO nuclear translocation to exert its transcriptional action. Further RNAi and EMSA analysis revealed that TcFOXO negatively regulated the expression of TcFDH. These results indicate that S6K might regulate beetles' reproduction through FOXO/JH signaling pathway.


Assuntos
Besouros , Tribolium , Animais , Besouros/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Hormônios Juvenis/metabolismo , Masculino , Óvulo/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , Tribolium/genética , Tribolium/metabolismo
16.
Sci Rep ; 12(1): 10905, 2022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35764778

RESUMO

Brassicaceae plants contain glucosinolates, which are hydrolysed by myrosinases to toxic products such as isothiocyanates and nitriles, acting as defences. Herbivores have evolved various detoxification strategies, which are reviewed here. Larvae of Phaedon cochleariae (Coleoptera: Chrysomelidae) metabolise hydrolysis products of benzenic glucosinolates by conjugation with aspartic acid. In this study, we investigated whether P. cochleariae uses the same metabolic pathway for structurally different glucosinolates, whether the metabolism differs between adults and larvae and which hydrolysis products are formed as intermediates. Feeding experiments were performed with leaves of watercress (Nasturtium officinale, Brassicaceae) and pea (Pisum sativum, non-Brassicaceae), to which glucosinolates with structurally different side chains (benzenic, indole or aliphatic) or their hydrolysis products were applied. Samples were analysed by UHPLC-QTOF-MS/MS or TD-GC-MS. The same aspartic acid conjugates as previously identified in larvae were also detected as major metabolites of benzenic glucosinolates in adults. Indol-3-ylmethyl glucosinolate was mainly metabolised to N-(1H-indol-3-ylcarbonyl) glutamic acid in adults and larvae, while the metabolism of 2-propenyl glucosinolate remains unclear. The metabolism may thus proceed primarily via isothiocyanates rather than via nitriles, while the hydrolysis occurs independently of plant myrosinases. A detoxification by conjugation with these amino acids is not yet known from other Brassicaceae-feeders.


Assuntos
Brassicaceae , Besouros , Nasturtium , Animais , Ácido Aspártico/metabolismo , Brassicaceae/metabolismo , Besouros/metabolismo , Glucosinolatos/metabolismo , Isotiocianatos/metabolismo , Larva/metabolismo , Nitrilas/metabolismo , Espectrometria de Massas em Tandem
17.
Environ Entomol ; 51(4): 700-709, 2022 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-35666204

RESUMO

The cigarette beetle, Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae), is a destructive stored product pest worldwide. Adult cigarette beetles are known to rely on host volatiles and pheromones to locate suitable habitats for oviposition and mating, respectively. However, little is known about the chemosensory mechanisms of these pests. Soluble chemoreception proteins are believed to initiate olfactory signal transduction in insects, which play important roles in host searching and mating behaviors. In this study, we sequenced the antennal transcriptome of L. serricorne and identified 14 odorant-binding proteins (OBPs), 5 chemosensory proteins (CSPs), and 2 Niemann-Pick C2 proteins (NPC2). Quantitative realtime PCR (qPCR) results revealed that several genes (LserOBP2, 3, 6, and 14) were predominantly expressed in females, which might be involved in specific functions in this gender. The five LserOBPs (LserOBP1, 4, 8, 10, and 12) that were highly expressed in the male antennae might encode proteins involved in specific functions in males. These findings will contribute to a better understanding of the olfactory system in this stored product pest and will assist in the development of efficient and environmentally friendly strategies for controlling L. serricorne.


Assuntos
Besouros , Receptores Odorantes , Animais , Antenas de Artrópodes/metabolismo , Besouros/genética , Besouros/metabolismo , Feminino , Perfilação da Expressão Gênica , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Masculino , Filogenia , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Transcriptoma
18.
Microb Genom ; 8(6)2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35678705

RESUMO

Plastics are inexpensive and widely used organic polymers, but their high durability hinders biodegradation. Polystyrene, including extruded polystyrene (also known as styrofoam), is among the most commonly produced plastics worldwide and is recalcitrant to microbial degradation. In this study, we assessed changes in the gut microbiome of superworms (Zophobas morio) reared on bran, polystyrene or under starvation conditions over a 3 weeks period. Superworms on all diets were able to complete their life cycle to pupae and imago, although superworms reared on polystyrene had minimal weight gains, resulting in lower pupation rates compared to bran reared worms. The change in microbial gut communities from baseline differed considerably between diet groups, with polystyrene and starvation groups characterized by a loss of microbial diversity and the presence of opportunistic pathogens. Inferred microbial functions enriched in the polystyrene group included transposon movements, membrane restructuring and adaptations to oxidative stress. We detected several encoded enzymes with reported polystyrene and styrene degradation abilities, supporting previous reports of polystyrene-degrading bacteria in the superworm gut. By recovering metagenome-assembled genomes (MAGs) we linked phylogeny and functions and identified genera including Pseudomonas, Rhodococcus and Corynebacterium that possess genes associated with polystyrene degradation. In conclusion, our results provide the first metagenomic insights into the metabolic pathways used by the gut microbiome of superworms to degrade polystyrene. Our results also confirm that superworms can survive on polystyrene feed, but this diet has considerable negative impacts on host gut microbiome diversity and health.


Assuntos
Besouros , Microbiota , Animais , Besouros/metabolismo , Larva/metabolismo , Microbiota/genética , Plásticos/metabolismo , Poliestirenos/metabolismo
19.
Pestic Biochem Physiol ; 184: 105129, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35715067

RESUMO

Callosobruchus maculatus is the main pest cowpea (Vigna unguiculata). Given its relevance as an insect pest, studies have focused in finding toxic compounds which could prevent its predatory action towards the seeds. Clitoria fairchildiana is a native Amazon species, whose seeds are refractory to insect predation. This characteristic was the basis of our interest in evaluating the toxicity of its seed proteins to C. maculatus larvae. Seed proteins were fractioned, according to their solubility, to albumins (F1), globulins (F2), kaphyrins (F3), glutelins (F4), linked kaphyrins (F5) and cross-linked glutelins (F6). The fractionated proteins were quantified, analysed by tricine-SDS-PAGE and inserted into the diet of this insect pest in order to evaluate their insecticidal potential. The most toxic fraction to C. maculatus, the propanol soluble F3, was submitted to molecular exclusion chromatography and all of the peaks obtained, F3P1, F3P2, F3P3, caused a reduction of larval mass, especially F3P1, seen as a major ~12 kDa electrophoretic band. This protein was identified as a vicilin-like protein by mass spectrometry and BLAST analysis. The alignment of the Cfvic (C. fairchildiana vicilin) peptides with a V. unguiculata vicilin sequence, revealed that Cfvic has at least five peptides (ALLTLVNPDGR, AILTLVNPDGR, NFLAGGKDNV, ISDINSAMDR, NFLAGEK) which lined up with two chitin binding sites (ChBS). This finding was corroborated by chitin affinity chromatography and molecular docking of chitin-binding domains for N-Acetyl-D-glucosamine and by the reduction of Cfvic chitin affinity after chemical modification of its Lys residues. In conclusion, Cfvic is a 12 kDa vicilin-like protein, highly toxic to C. maculatus, acting as an insect toxin through its ability to bind to chitin structures present in the insect midgut.


Assuntos
Clitoria , Besouros , Animais , Quitina/metabolismo , Clitoria/metabolismo , Besouros/metabolismo , Cotilédone/metabolismo , Glutens/análise , Glutens/metabolismo , Larva/metabolismo , Simulação de Acoplamento Molecular , Proteínas de Armazenamento de Sementes , Sementes/química
20.
Pest Manag Sci ; 78(8): 3508-3518, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35576327

RESUMO

BACKGROUND: Insect cytochrome P450 monooxygenases (P450s) play a crucial role in phytochemical metabolism and tolerance. Three P450 genes (TcCYP4Q3, TcCYP4Q5, and TcCYP4Q7) are associated with the response of eugenol in Tribolium castaneum. However, the responding mechanisms of these P450 genes to eugenol remain unknown. RESULTS: Here, spatiotemporal expression profiling revealed that TcCYP4Q3 and TcCYP4Q5 were most highly expressed in late adult, while TcCYP4Q7 was predominantly expressed in late larva; and all of these three P450 genes were mainly expressed in the fat body of larvae. Furthermore, the expressions of these three P450 genes were significantly up-regulated after exposure to eugenol, and depletion of them enhanced the susceptibility of beetles to eugenol. Interestingly, RNA interference (RNAi) against the CncC gene, a transcription factor of CncC signaling pathway associated with regulation of insect P450s in response to phytochemicals, reduced the transcripts of these three P450 genes following exposure to eugenol. Investigation of CncC signaling pathway showed that this pathway could be activated by eugenol. CONCLUSION: Altogether, the results indicate that these three P450 genes are regulated by CncC signaling pathway to participate in the susceptibility of Tribolium castaneum to phytochemicals. These findings will aid implications for the development of novel therapeutics to control pest. © 2022 Society of Chemical Industry.


Assuntos
Besouros , Tribolium , Animais , Besouros/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Família 4 do Citocromo P450/genética , Família 4 do Citocromo P450/metabolismo , Eugenol , Larva/genética , Larva/metabolismo , Compostos Fitoquímicos , Interferência de RNA , Transdução de Sinais , Tribolium/genética , Tribolium/metabolismo
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