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1.
Anal Chem ; 92(21): 14730-14739, 2020 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-33064451

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to a global pandemic of coronavirus disease 2019 (COVID-19). The spike protein expressed on the surface of this virus is highly glycosylated and plays an essential role during the process of infection. We conducted a comprehensive mass spectrometric analysis of the N-glycosylation profiles of the SARS-CoV-2 spike proteins using signature ions-triggered electron-transfer/higher-energy collision dissociation (EThcD) mass spectrometry. The patterns of N-glycosylation within the recombinant ectodomain and S1 subunit of the SARS-CoV-2 spike protein were characterized using this approach. Significant variations were observed in the distribution of glycan types as well as the specific individual glycans on the modification sites of the ectodomain and subunit proteins. The relative abundance of sialylated glycans in the S1 subunit compared to the full-length protein could indicate differences in the global structure and function of these two species. In addition, we compared N-glycan profiles of the recombinant spike proteins produced from different expression systems, including human embryonic kidney (HEK 293) cells and Spodoptera frugiperda (SF9) insect cells. These results provide useful information for the study of the interactions of SARS-CoV-2 viral proteins and for the development of effective vaccines and therapeutics.


Assuntos
Betacoronavirus/química , Polissacarídeos/análise , Glicoproteína da Espícula de Coronavírus/química , Animais , Glicosilação , Células HEK293 , Humanos , Espectrometria de Massas/métodos , Polissacarídeos/química , Spodoptera/química
2.
Nat Commun ; 11(1): 4916, 2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-33004803

RESUMO

Self-incompatibility (SI) is a breeding system that promotes cross-fertilization. In Brassica, pollen rejection is induced by a haplotype-specific interaction between pistil determinant SRK (S receptor kinase) and pollen determinant SP11 (S-locus Protein 11, also named SCR) from the S-locus. Although the structure of the B. rapa S9-SRK ectodomain (eSRK) and S9-SP11 complex has been determined, it remains unclear how SRK discriminates self- and nonself-SP11. Here, we uncover the detailed mechanism of self/nonself-discrimination in Brassica SI by determining the S8-eSRK-S8-SP11 crystal structure and performing molecular dynamics (MD) simulations. Comprehensive binding analysis of eSRK and SP11 structures reveals that the binding free energies are most stable for cognate eSRK-SP11 combinations. Residue-based contribution analysis suggests that the modes of eSRK-SP11 interactions differ between intra- and inter-subgroup (a group of phylogenetically neighboring haplotypes) combinations. Our data establish a model of self/nonself-discrimination in Brassica SI.


Assuntos
Brassica rapa/fisiologia , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Animais , Cristalografia , Flores/metabolismo , Haplótipos , Simulação de Dinâmica Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/ultraestrutura , Pólen/metabolismo , Ligação Proteica/fisiologia , Domínios Proteicos/fisiologia , Proteínas Quinases/genética , Proteínas Quinases/ultraestrutura , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/ultraestrutura , Células Sf9 , Spodoptera
3.
Zootaxa ; 4772(2): zootaxa.4772.2.11, 2020 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-33055621

RESUMO

The neotropical fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) is an important lepidopteran pest with a broad geographical distribution (Goergen et al. 2015, Early et al. 2018, Sharanabasappa et al. 2018). Larvae of S. frugiperda feed on more than 350 plant species, including several economically-important crops such as maize, sugarcane or rice (Montezano et al. 2018). Following its invasion of the African continent and of South Asia in 2016 and 2018 respectively, FAW has caused important crop losses and associated livelihood impacts (Baudron et al., 2019). During 2019, FAW rapidly spread across Southeast Asia and FAW-attributed feeding damage was recorded on maize in Viet Nam. In Viet Nam, maize is the second most important staple food crop after rice, is extensively used for livestock production and poultry feed, and thus plays a central role in sustaining rural livelihoods (Dang et al. 2004). Hence, the recent FAW invasion and the anticipated pest-induced yield losses will carry important repercussions for local maize value chains and are likely to degrade farmers' revenue base. In this study, we conducted a morphological and molecular identification of locally-collected FAW individuals to better characterize the species' invasion history in Viet Nam.


Assuntos
Spodoptera , Animais , Oryza , Vietnã , Zea mays
4.
Emerg Microbes Infect ; 9(1): 2076-2090, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32897177

RESUMO

The current coronavirus disease 2019 (COVID-19) pandemic was the result of the rapid transmission of a highly pathogenic coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which there is no efficacious vaccine or therapeutic. Toward the development of a vaccine, here we expressed and evaluated as potential candidates four versions of the spike (S) protein using an insect cell expression system: receptor binding domain (RBD), S1 subunit, the wild-type S ectodomain (S-WT), and the prefusion trimer-stabilized form (S-2P). We showed that RBD appears as a monomer in solution, whereas S1, S-WT, and S-2P associate as homotrimers with substantial glycosylation. Cryo-electron microscopy analyses suggested that S-2P assumes an identical trimer conformation as the similarly engineered S protein expressed in 293 mammalian cells but with reduced glycosylation. Overall, the four proteins confer excellent antigenicity with convalescent COVID-19 patient sera in enzyme-linked immunosorbent assay (ELISA), yet show distinct reactivities in immunoblotting. RBD, S-WT and S-2P, but not S1, induce high neutralization titres (>3-log) in mice after a three-round immunization regimen. The high immunogenicity of S-2P could be maintained at the lowest dose (1 µg) with the inclusion of an aluminium adjuvant. Higher doses (20 µg) of S-2P can elicit high neutralization titres in non-human primates that exceed 40-times the mean titres measured in convalescent COVID-19 subjects. Our results suggest that the prefusion trimer-stabilized SARS-CoV-2 S-protein from insect cells may offer a potential candidate strategy for the development of a recombinant COVID-19 vaccine.


Assuntos
Antígenos Virais/imunologia , Betacoronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Imunogenicidade da Vacina/imunologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Glicoproteína da Espícula de Coronavírus/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Linhagem Celular , Infecções por Coronavirus/imunologia , Microscopia Crioeletrônica , Ensaio de Imunoadsorção Enzimática , Humanos , Macaca fascicularis , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização , Peptidil Dipeptidase A/metabolismo , Domínios Proteicos/genética , Domínios Proteicos/imunologia , Células Sf9 , Glicoproteína da Espícula de Coronavírus/genética , Spodoptera , Vacinação , Proteínas do Envelope Viral/imunologia
5.
PLoS One ; 15(9): e0238523, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32911522

RESUMO

Brazil and South Africa, countries with economies in transition, are characterized by a dual agrarian structure co-occurring, sometimes, alongside in the same region. Large-scale commercial farming produces crops for export to global markets on the one hand, and small-scale farming, on the other hand, provides for subsistence and local markets. In both systems, maize (Zea mays) is a key crop for these two countries. For the commercial system, maize is a commodity crop while for the small-scale system it is the prime staple crop. In commercial systems, farmers predominantly grow genetically modified (GM) hybrid maize. In small-scale systems, farmers rely on open pollinated varieties (OPVs) and/or landraces and are largely dependent on seed saving systems. The aim of this study was to understand the relationship between transgene expression rates, the resulting concentrations of the transgene product (Bt protein) and its bioactivity in insect pests following transgene flow from GM hybrid maize into non-genetically modified, non-GM near-isogenic maize hybrid (ISO) and OPVs. We modeled segregation patterns and measured cry1Ab transgene expression (mRNA quantification), Cry1Ab protein concentration and pest survival. Two groups of F1, F2 crosses and backcrosses with GM, ISO and OPV maize varieties from Brazil and South Africa were used. Bioassays with the larvae of two lepidopteran maize pest species, Helicoverpa armigera and Spodoptera littoralis, were carried out. Overall, the cry1Ab transgene outcrossed effectively into the genetic backgrounds tested. The cry1Ab transgene was stably expressed in both ISO and OPV genetic backgrounds. Transgene introgression led to consistent, though highly variable, concentrations of Cry1Ab toxins that were similar to those observed in GM parental maize. Most crosses, but not all, suggested the expected Mendelian segregation pattern. Transgene expression rates were significantly higher than expected from purely Mendelian segregation in the South African crosses. In South African materials, ISO and OPV crosses produced significantly lower Cry1Ab concentrations compared to the GM parental maize. The Cry1Ab toxins from crosses were bioactive and induced mortality rates of ≥92.19% in H. armigera and ≥40.63% in S. littoralis after a period of only 4 days. However, no correlations were observed between the quantitation of mRNA for cry1Ab and the corresponding Cry1Ab protein concentrations, nor between the Cry1Ab concentrations and insect mortality rates across different genetic backgrounds. We therefore suggest that while transcription of the cry1Ab transgene reliably determines the presence of Cry1Ab protein, mRNA levels do not reflect, by themselves, the end Cry1Ab protein concentrations found in the plant. Because predictably high Cry1Ab concentrations are a key prerequisite for effective insect resistance management (IRM) programs, this observation raises questions about the effectiveness of such programs in scenarios with complex crop genetic backgrounds. On the other hand, confirmed bioactivity in all crosses should be expected to impact small farmer's selection behavior, unknowingly favoring the insecticidal trait. This may lead to a fixation of the trait in the wider population, and may influence the genetic diversity of varieties maintained by small-scale farmers.


Assuntos
Proteínas de Bactérias/genética , Endotoxinas/genética , Proteínas Hemolisinas/genética , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Zea mays/genética , Animais , Produtos Agrícolas/genética , Cruzamentos Genéticos , Resistência à Doença , Doenças das Plantas/genética , Doenças das Plantas/parasitologia , Plantas Geneticamente Modificadas/parasitologia , Spodoptera/fisiologia , Transgenes , Zea mays/parasitologia
6.
Viruses ; 12(9)2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32971895

RESUMO

Coronaviruses (CoVs) are enveloped, positive sense, single strand RNA viruses that cause respiratory, intestinal and neurological diseases in mammals and birds. Following replication, CoVs assemble on intracellular membranes including the endoplasmic reticulum Golgi intermediate compartment (ERGIC) where the envelope protein (E) functions in virus assembly and release. In consequence, E potentially contains membrane-modifying peptides. To search for such peptides, the E coding sequence of Mouse Hepatitis Virus (MHV) was inspected for its amino acid conservation, proximity to the membrane and/or predicted amphipathic helices. Peptides identified in silico were synthesized and tested for membrane-modifying activity in the presence of giant unilamellar vesicles (GUVs) consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), sphingomyelin and cholesterol. To confirm the presence of membrane binding peptides identified in the context of a full-length E protein, the wild type and a number of mutants in the putative membrane binding peptide were expressed in Lenti-X-293T mammalian and insect cells, and the distribution of E antigen within the expressing cell was assessed. Our data identify a role for the post-transmembrane region of MHV E in membrane binding.


Assuntos
Vírus da Hepatite Murina/química , Peptídeos/química , Proteínas do Envelope Viral/química , Sequência de Aminoácidos , Animais , Linhagem Celular , Infecções por Coronavirus , Humanos , Membranas Intracelulares/metabolismo , Camundongos , Vírus da Hepatite Murina/genética , Vírus da Hepatite Murina/metabolismo , Mutação , Peptídeos/síntese química , Peptídeos/metabolismo , Células Sf9 , Spodoptera , Lipossomas Unilamelares/metabolismo , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo
7.
Sci Total Environ ; 740: 140015, 2020 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-32927537

RESUMO

Fall armyworm (FAW) is a new invasive pest that is causing devastating effects on maize production and threatening the livelihoods of millions of poor smallholders across sub-Saharan Africa and Asia. Using unique survey data from 2356 maize-growing households in Ghana, Rwanda, Uganda, Zambia and Zimbabwe, we examined how smallholder farmers are fighting this voracious pest. In particular, we assessed the FAW management strategies used by smallholders, socio-economic factors driving the choice of the management options, the complementarities or tradeoffs among the management options, and the (un)safe pesticide use practices of farmers. Results showed that smallholder farm households have adopted a variety of cultural, physical, chemical and local options to mitigate the effects of FAW, but the use of synthetic pesticides remains the most popular option. Results from multivariate probit regressions indicated that the extensive use of synthetic pesticides is driven by household asset wealth, and access to subsidised farm inputs and extension information. We observed that farm households are using a wide range of pesticides, including highly hazardous and banned products. Unfortunately, a majority of the households do not use personal protective equipment while handling the pesticides, resulting in reports of acute pesticide-related illness. Our findings have important implications for policies and interventions aimed at promoting environmentally friendly and sustainable ways of managing invasive pests in smallholder farming systems.


Assuntos
Espécies Introduzidas , Praguicidas , Spodoptera , Agricultura , Animais , Ásia , Gana , Ruanda , Uganda , Zâmbia , Zimbábue
8.
Nat Commun ; 11(1): 4160, 2020 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-32814772

RESUMO

Ghrelin is a gastric peptide hormone with important physiological functions. The unique feature of ghrelin is its Serine 3 acyl-modification, which is essential for ghrelin's activity. However, it remains to be elucidated why the acyl-modification of ghrelin is necessary for activity. To address these questions, we solved the crystal structure of the ghrelin receptor bound to antagonist. The ligand-binding pocket of the ghrelin receptor is bifurcated by a salt bridge between E124 and R283. A striking feature of the ligand-binding pocket of the ghrelin receptor is a wide gap (crevasse) between the TM6 and TM7 bundles that is rich in hydrophobic amino acids, including a cluster of phenylalanine residues. Mutagenesis analyses suggest that the interaction between the gap structure and the acyl acid moiety of ghrelin may participate in transforming the ghrelin receptor into an active conformation.


Assuntos
Grelina/metabolismo , Fenilalanina/metabolismo , Receptores de Grelina/metabolismo , Animais , Sítios de Ligação/genética , Células CHO , Cricetinae , Cricetulus , Cristalografia por Raios X , Grelina/química , Grelina/genética , Células HEK293 , Humanos , Ligantes , Camundongos Endogâmicos MRL lpr , Mutagênese Sítio-Dirigida , Fenilalanina/química , Fenilalanina/genética , Ligação Proteica , Conformação Proteica , Receptores de Grelina/antagonistas & inibidores , Receptores de Grelina/genética , Células Sf9 , Spodoptera
9.
Ecotoxicol Environ Saf ; 202: 110935, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32800218

RESUMO

This study investigated the ability of dual crosslinked interpenetrating polymer network (IPN) blend beads (DIN:SA/PVA-beads), composed of sodium alginate (SA) and poly (vinyl alcohol) (PVA), as a base-triggered carrier for the controlled release of dinotefuran (DIN) in Spodoptera litera midgut. The blend beads were characterized for morphology, encapsulation efficiency, swelling degree, and in vitro release of the blend beads were characterized. The results revealed that the double-crosslinked gel beads had a tightly interpenetrating network structure and exhibited a satisfactory embedding effect for DIN. The maximum of the DIN loading capacity was approximately 1.01%, with a high encapsulation efficiency of 83.19%. The triggered release of DIN from the blend beads was studied in deionized water (pH 3.0-11.0) via high-performance liquid chromatography (HPLC); it was found that the release rate was higher in alkaline pH conditions than in acidic and neutral conditions. An in vivo dynamics and degradation study also demonstrated that the excellent release characteristics of DIN:SA/PVA-beads in the midgut of S. litera. This study provides a promising controlled-release form of dinotefuran that is more effective and can be used for the targeted control of pests with alkaline midgut.


Assuntos
Guanidinas/metabolismo , Neonicotinoides/metabolismo , Nitrocompostos/metabolismo , Spodoptera/metabolismo , Alginatos/química , Animais , Preparações de Ação Retardada/química , Etanol , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Hidrogéis/química , Concentração de Íons de Hidrogênio , Polímeros , Álcool de Polivinil/química
10.
PLoS Biol ; 18(8): e3000790, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32776918

RESUMO

Concentrative nucleoside transporters (CNTs), members of the solute carrier (SLC) 28 transporter family, facilitate the salvage of nucleosides and therapeutic nucleoside derivatives across the plasma membrane. Despite decades of investigation, the structures of human CNTs remain unknown. We determined the cryogenic electron microscopy (cryo-EM) structure of human CNT (hCNT) 3 at an overall resolution of 3.6 Å. As with its bacterial homologs, hCNT3 presents a trimeric architecture with additional N-terminal transmembrane helices to stabilize the conserved central domains. The conserved binding sites for the substrate and sodium ions unravel the selective nucleoside transport and distinct coupling mechanism. Structural comparison of hCNT3 with bacterial homologs indicates that hCNT3 is stabilized in an inward-facing conformation. This study provides the molecular determinants for the transport mechanism of hCNTs and potentially facilitates the design of nucleoside drugs.


Assuntos
Proteínas de Bactérias/química , Proteínas de Membrana Transportadoras/química , Uridina/química , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Baculoviridae/genética , Baculoviridae/metabolismo , Sítios de Ligação , Transporte Biológico , Clonagem Molecular , Microscopia Crioeletrônica , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Modelos Moleculares , Ligação Proteica , Conformação Proteica em alfa-Hélice , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Células Sf9 , Spodoptera , Homologia Estrutural de Proteína , Especificidade por Substrato , Uridina/metabolismo
11.
PLoS One ; 15(8): e0236759, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32745105

RESUMO

The fall armyworm (Spodoptera frugiperda) is a moth pest native to the Western Hemisphere that has recently become a global problem, invading Africa, Asia, and Australia. The species has a broad host range, long-distance migration capability, and a propensity for the generation of pesticide resistance traits that make it a formidable invasive threat and a difficult pest to control. While fall armyworm migration has been extensively studied in North America, where annual migrations of thousands of kilometers are the norm, migration patterns in South America are less understood. As a first step to address this issue we have been genetically characterizing fall armyworm populations in Ecuador, a country in the northern portion of South America that has not been extensively surveyed for this pest. These studies confirm and extend past findings indicating similarities in the fall armyworm populations from Ecuador, Trinidad-Tobago, Peru, and Bolivia that suggest substantial migratory interactions. Specifically, we found that populations throughout Ecuador are genetically homogeneous, indicating that the Andes mountain range is not a long-term barrier to fall armyworm migration. Quantification of genetic variation in an intron sequence describe patterns of similarity between fall armyworm from different locations in South America with implications for how migration might be occurring. In addition, we unexpectedly found these observations only apply to one subset of fall armyworm (the C-strain), as the other group (R-strain) was not present in Ecuador. The results suggest differences in migration behavior between fall armyworm groups in South America that appear to be related to differences in host plant preferences.


Assuntos
Haplótipos/genética , Spodoptera/genética , Migração Animal , Animais , Equador , Complexo IV da Cadeia de Transporte de Elétrons/genética , Marcadores Genéticos , Íntrons/genética , Controle de Pragas , Filogenia , Filogeografia , América do Sul
12.
PLoS One ; 15(8): e0237258, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32756605

RESUMO

Aquatic weeds such as muskgrass (Chara spp.), water hyacinth (Eichhornia crassipes), water lettuce (Pistia stratiotes), hydrilla (Hydrilla verticillate), filamentous algae (Lyngbya wollei), and duckweed (Lemna minor) thrive in farm canals within the Everglades Agricultural Area of South Florida. Their presence, particularly during the summer months is an environmental concern with regards to water quality, in addition to being a nuisance because of their ability to multiply and spread rapidly in open waters causing restricted drainage/irrigation flow and low dissolved oxygen levels. Chemical control is effective but can have undesirable off-target effects, so reduced herbicide use is desirable. Hence, need exists to discover ways in which these weeds could be best managed or utilized. The objective of this research was to evaluate the allelopathic effect of these weeds to determine their use as potential biopesticides. Six aqueous extracts were tested against 100 bacterial strains isolated from plants and soil to evaluate their antimicrobial activity. These extracts were also used to determine their insecticidal and antifeedant effects on fall armyworm (FAW, Spodoptera frugiperda). Both extracts and powder form of the aquatic weeds were tested for their herbicidal activity towards seed germination and growth of three common terrestrial weed species. At a dilution of 1:100 and 1:1,000, none of the aquatic weeds inhibited in-vitro growth of the bacterial strains, with one exception (filamentous algae extract at 1:100 reduced growth of one bacterial isolate by 54%). Water lettuce reduced the survival rate of FAW by 14% while hydrilla and duckweed caused 11% and 9% reduction of FAW growth, respectively. Powdered duckweed inhibited the growth of nutsedge by 41%, whereas filamentous algae powder and extract reduced germination of amaranth by 20% and 28%, respectively. Harvesting these weeds and converting them into useable compounds could not only eliminate the in situ farm canal and water quality problems but also result in development of new soil amendments or biopesticides.


Assuntos
Antibacterianos/toxicidade , Agentes de Controle Biológico/toxicidade , Plantas Daninhas/química , Agricultura , Animais , Antibacterianos/química , Bactérias/efeitos dos fármacos , Agentes de Controle Biológico/química , Cianobactérias/química , Eichhornia/química , Hydrocharitaceae/química , Inseticidas/química , Inseticidas/toxicidade , Plantas/microbiologia , Microbiologia do Solo , Spodoptera/efeitos dos fármacos
13.
Ecotoxicol Environ Saf ; 204: 111052, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32739675

RESUMO

Transgenerational effects on sensitivity to pesticides are poorly studied. This study investigated the transgenerational influences of maternal body mass in the major pest moth Spodoptera littoralis, with a focus on sensitivity to chlorpyrifos pesticide. In 147 clutches of a laboratory strain of S. littoralis, we compared larval mortality between control larvae and larvae treated with chlorpyrifos. Because of the classic positive relationships between offspring size and maternal size and between offspring size and offspring quality, sensitivity to chlorpyrifos was predicted to be lower in larvae of larger mothers. Surprisingly, we found the opposite result, with higher pesticide toxicity in larvae of larger mothers. This result is partly explained by the lack of a relationship between larval mass and larval sensitivity to chlorpyrifos. This means that another offspring characteristic linked to maternal size should have affected larval sensitivity to chlorpyrifos. More generally, knowledge of the effects of the traits and ecological environments of mothers on offspring sensitivity to pesticides remains limited. Ecotoxicologists should pay more attention to such maternal effects on sensitivity to pesticides, both in pests and non-target species.


Assuntos
Clorpirifos/toxicidade , Epigênese Genética , Resistência a Inseticidas/genética , Inseticidas/toxicidade , Spodoptera/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Feminino , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento , Longevidade/efeitos dos fármacos , Exposição Materna , Spodoptera/genética , Spodoptera/crescimento & desenvolvimento
14.
Sci Total Environ ; 745: 141048, 2020 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-32758757

RESUMO

Human activity is a serious cause of extensive changes in the environment and a constant reason for the emergence of new stress factors. Thus, to survive and reproduce, organisms must constantly implement a program of adaptation to continuously changing conditions. The research presented here is focused on tracking slow changes occurring in Spodoptera exigua (Lepidoptera: Noctuidae) caused by multigenerational exposure to sub-lethal cadmium doses. The insects received food containing cadmium at concentrations of 5, 11, 22 and 44 µg per g of dry mass of food. The level of DNA stability was monitored by a comet assay in subsequent generations up to the 36th generation. In the first three generations, the level of DNA damage was high, especially in the groups receiving higher doses of cadmium in the diet. In the fourth generation, a significant reduction in the level of DNA damage was observed, which could indicate that the desired stability of the genome was achieved. Surprisingly, however, in subsequent generations, an alternating increase and decrease was found in DNA stability. The observed cycles of changing DNA stability were longer lasting in insects consuming food with a lower Cd content. Thus, a transient reduction in genome stability can be perceived as an opportunity to increase the number of genotypes that undergo selection. This phenomenon occurs faster if the severity of the stress factor is high but is low enough to allow the population to survive.


Assuntos
Cádmio/toxicidade , Dano ao DNA , Animais , Instabilidade Genômica , Humanos , Larva , Spodoptera/genética
15.
Pestic Biochem Physiol ; 169: 104649, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32828367

RESUMO

Spodoptera litura is a destructive agricultural pest and has evolved resistance to multiple insecticides, especially pyrethroids. At present, the resistance mechanism to pyrethroids remains unclear. Four field-collected populations, namely CZ, LF, NJ and JD, were identified to have high resistance to pyrethroids comparing to pyrethroid-susceptible population (GX), with resistant ratio ranging from 11.5- to 9123.5-fold. To characterize pyrethroid resistance mechanism, the transcriptomes between two pyrethroid-resistant (LF and NJ) and a pyrethroid-susceptible (GX) populations were compared by RNA-sequencing. Results showed that multiple differentially expressed genes were enriched in metabolism-related GO terms and KEGG pathways. 35 up-regulated metabolism-related unigenes were selected to verify by qRT-PCR and 15 unigenes, including 4 cytochrome P450s (P450s), 5 glutathione S-transferase (GSTs), 1 UDP-glycosyltransferase (UGT), 4 carboxylesterases (COEs) and 1 and ATP-binding cassette transporters (ABC), were all up-regulated in the four pyrethroid-resistant populations. The expression levels of CYP3 and GST3, which were annotated as CYP6A13 and GSTE1, respectively, showed positive correlation with their pyrethroid resistance levels among the four pyrethroid-resistant populations. While the expression levels of CYP5, CYP12, COE4 and ABC5 showed good correlation with their pyrethroid resistance levels in at least three populations. UGT5 had the highest expression level among the tested UGT genes in the four pyrethroid-resistant populations. RNAi mediated silencing of CYP6 increased the cumulative mortality treated by beta cypermethrin and cyhalothrin significantly, while silencing of GST3 increased the cumulative mortality treated by fenvalerate significantly. CYP3, CYP5, CYP6, CYP12, GST3, COE4, UGT5 and ABC5 play important roles in pyrethroid resistance among the four pyrethroid-resistant populations. Our work provides a valuable clue for further study of pyrethroid resistance mechanisms in S. litura.


Assuntos
Inseticidas/farmacologia , Piretrinas , Animais , Perfilação da Expressão Gênica , Resistência a Inseticidas/efeitos dos fármacos , Spodoptera/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos
16.
Pestic Biochem Physiol ; 169: 104652, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32828370

RESUMO

Neurophysiological recordings were employed to quantify neuronal sensitivity to neurotoxic insecticides and assessed toxicity across field and laboratory fall armyworm (FAW) populations. Topical toxicity resistance ratios (RR) in field-collected FAW was 767-fold compared to laboratory strains and, importantly, a 1750-fold reduction in potency was observed for λ-cyhalothrin in neurophysiological assays. Field collected FAW were found to have a RR of 12 to chlorpyrifos when compared to the susceptible strain and was 8-fold less sensitive in neurophysiological assays. Surprisingly, there were no point mutations identified in the voltage-gated sodium channel known to cause pyrethroid resistance. For acetylcholinesterase, FAW had more than 80% of their nucleotide sequences consistent with A201 and F290 of the susceptible strains although 60% of the tested population was heterozygous for the G227A mutation. These data indicate that point mutations did not contribute to the high level of pyrethroid resistance and nerve insensitivity in this population of field collected FAW. Additionally, these data suggest the kdr phenotype only explains a portion of the heritable variation in FAW resistance and indicates kdr is not the only predictor of high pyrethroid resistance. Phenotypic assays, such as toxicity bioassays or neurophysiological recordings, using field-collected populations are necessary to reliably predict resistant phenotypes and product failures.


Assuntos
Inseticidas/farmacologia , Piretrinas , Animais , Resistência a Inseticidas/efeitos dos fármacos , Mutação , Spodoptera/efeitos dos fármacos
17.
Proc Natl Acad Sci U S A ; 117(35): 21403-21412, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32817418

RESUMO

The early steps of DNA double-strand break (DSB) repair in human cells involve the MRE11-RAD50-NBS1 (MRN) complex and its cofactor, phosphorylated CtIP. The roles of these proteins in nucleolytic DSB resection are well characterized, but their role in bridging the DNA ends for efficient and correct repair is much less explored. Here we study the binding of phosphorylated CtIP, which promotes the endonuclease activity of MRN, to single long (∼50 kb) DNA molecules using nanofluidic channels and compare it to the yeast homolog Sae2. CtIP bridges DNA in a manner that depends on the oligomeric state of the protein, and truncated mutants demonstrate that the bridging depends on CtIP regions distinct from those that stimulate the nuclease activity of MRN. Sae2 is a much smaller protein than CtIP, and its bridging is significantly less efficient. Our results demonstrate that the nuclease cofactor and structural functions of CtIP may depend on the same protein population, which may be crucial for CtIP functions in both homologous recombination and microhomology-mediated end-joining.


Assuntos
Quebras de DNA de Cadeia Dupla , DNA Circular/metabolismo , Endodesoxirribonucleases/metabolismo , Animais , Endonucleases/metabolismo , Humanos , Nanotecnologia , Fosforilação , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomycetales , Células Sf9 , Spodoptera
18.
Pestic Biochem Physiol ; 168: 104623, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32711763

RESUMO

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is the main destructive insect pest of grain crops that occurs in all maize growing regions of the Americas. It has rapidly invaded the Southern China since January 2019. However, the current status of insecticide resistance in S. frugiperda has not been reported in China. In this study, we determined the susceptibility of eight populations of FAW to eight insecticides by an artificial diet incorporation method. The results showed that among eight insecticides, emamectin benzoate, spinetoram, chlorantraniliprole, chlorfenapyr, and lufenuron showed higher toxicity to this pest, while lambda-cyhalothrin and azadirachtin exhibited lower toxicity. Susceptibility of S. frugiperda to indoxacarb was significantly different (10.0-fold for LC50) across the various geographic populations. To investigate the biochemical mechanism of FAW to lambda-cyhalothrin, we performed the synergism tests and the results showed that piperonyl butoxide (PBO) and triphenyl phosphate (TPP) produced a high synergism of lambda-cyhalothrin effects in the two field populations. Sequencing of the gene encoding the acetylcholinesterase (AChE) gene in the two field populations identified two amino acid mutations, all of which have been shown previously to confer resistance to organophosphates (OPs) in several arthropod species. The results of this study provided valuable information for choosing alternative insecticides and for insecticide resistance management of S. frugiperda.


Assuntos
Inseticidas/farmacologia , Animais , China , Resistência a Inseticidas/efeitos dos fármacos , Larva/efeitos dos fármacos , Nitrilos , Piretrinas , Spodoptera/efeitos dos fármacos
19.
Pestic Biochem Physiol ; 168: 104632, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32711766

RESUMO

Phytochemicals are toxic to insects, but their insecticidal efficiencies are usually low compared to synthetic insecticides. Understanding the mechanism of insect adaptation to phytochemicals will provide guidance for increasing their efficacy. Reduced glutathione (GSH) is a scavenger of reactive oxygen species (ROS) induced by phytochemicals. However, in insects, the pathway of GSH biosynthesis in response to phytochemicals is unclear. We found that exposure to 0.5% indole-3-methanol (I3C), xanthotoxin, and rotenone (ROT) significantly retarded the growth of Spodoptera litura larvae. The oxidative stress in S. litura larvae exposed to phytochemicals was increased. The up-regulation of glutamate cysteine ligase but not glutathione reductase revealed that the de novo synthesis pathway is responsible for GSH synthesis in phytochemical-treated larvae. Treatment with the inhibitor (BSO) of γ-glutamylcysteine synthetase (gclc), a subunit of glutamate cysteine ligase, resulted in decreases of GSH levels and GST activities, increases of ROS levels in I3C-treated larvae, which finally caused midgut necrosis and larval death. Treatment with BSO or I3C alone did not cause larval death. The addition of GSH could partly reduce the influence of I3C and BSO on S. litura growth. Nilaparvata lugens gclc RNAi confirmed the result of BSO treatment in S. litura. N. lugens gclc RNAi significantly increased the mortality of ROT-sprayed N. lugens, in which ROS levels were significantly increased. All data indicate that gclc is involved in insect response to phytochemical treatment. Treatment with dsgclc will increase the insecticidal efficacy of plant-derived compounds.


Assuntos
Vias Biossintéticas , Glutationa , Animais , Larva , Compostos Fitoquímicos , Spodoptera
20.
Environ Sci Pollut Res Int ; 27(34): 43103-43116, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32725570

RESUMO

Pest insects causing damage to cultivable crops and food products by feeding, fecundity, and parasitizing livestock, also being a nuisance to human health. In consideration with human health, the World Health Organization reports that more than 50% of the world's population is presently at risk from mosquito-borne diseases. Mosquitoes are primary vectors for major dreadful diseases such as yellow fever, malaria, and dengue fever, which infect millions of human beings all over the world and kill millions of peoples every year. The present research work was carried out to evaluate the antifeedant, larvicidal, pupicidal, larval, and pupal duration activity of Leonotis nepetifolia-mediated silver nanoparticles (AgNPs) against Spodoptera litura, Helicoverpa armigera, Aedes aegypti, and Culex quinquefasciatus. Biosynthesized AgNPs were characterized through various techniques such as UV-Vis spectrometer, X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta potential analysis. The AgNPs showed potential antifeedant activity of 78.77% and 82.16% against the larvae of S. litura and H. armigera, respectively. The maximum larval mortality rate (78.49% and 72.70%) and maximum pupal mortality rate (84.66% and 77.44%) were observed against S. litura and H. armigera. Mosquito larvae were tested with biosynthesized AgNPs, and recorded LC50 values were 47.44 ppm and 35.48 ppm on A. aegypti and C. quinquefasciatus, respectively. The histological examinations showed that the acceleration of the nanomaterial caused severe tissue damage in the epithelial and goblet cells in the larval midgut region of S. litura, H. armigera, A. aegypti, and C. quinquefasciatus. Biosynthesis of silver nanoparticles using L. nepetifolia is an ideal eco-friendly approach for the management of insect pests. Graphical abstract.


Assuntos
Aedes , Anopheles , Culex , Inseticidas , Lamiaceae , Nanopartículas Metálicas , Animais , Larva , Nanopartículas Metálicas/toxicidade , Mosquitos Vetores , Extratos Vegetais , Folhas de Planta , Prata , Spodoptera
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