RESUMO
One assumed function of herbivore-induced plant volatiles (HIPVs) is to attract natural enemies of the inducing herbivores. Field evidence for this is scarce. In addition, the assumption that elicitors in oral secretions that trigger the volatile emissions are essential for the attraction of natural enemies has not yet been demonstrated under field conditions. After observing predatory social wasps removing caterpillars from maize plants, we hypothesized that these wasps use HIPVs to locate their prey. To test this, we conducted an experiment that simultaneously explored the importance of caterpillar oral secretions in the interaction. Spodoptera caterpillars pinned onto mechanically damaged plants treated with oral secretion were more likely to be attacked by wasps compared with caterpillars on plants that were only mechanically wounded. Both of the latter treatments were considerably more attractive than plants only treated with oral secretion or left untreated. Subsequent analyses of headspace volatiles confirmed differences in emitted volatiles that likely account for the differential predation across treatments. These findings highlight the importance of HIPVs in prey localization by social wasps, hitherto underappreciated potential biocontrol agents and provide evidence for the role that elicitors play in inducing attractive odour blends.
Assuntos
Larva , Comportamento Predatório , Spodoptera , Compostos Orgânicos Voláteis , Vespas , Animais , Vespas/fisiologia , Compostos Orgânicos Voláteis/metabolismo , Spodoptera/fisiologia , Larva/fisiologia , Zea mays , HerbivoriaRESUMO
BACKGROUND: The fall armyworm (FAW, Spodoptera frugiperda) is a polyphagous pest known for causing significant crop damage. The gut microbiota plays a pivotal role in influencing the biology, physiology and adaptation of the host. However, understanding of the taxonomic composition and functional characteristics of the gut microbiota in FAW larvae fed on different host plants remains limited. METHODS: This study utilized metagenomic sequencing to explore the structure, function and antibiotic resistance genes (ARGs) of the gut microbiota in FAW larvae transferred from an artificial diet to four distinct host plants: maize, sorghum, tomato and pepper. RESULTS: The results demonstrated significant variations in gut microbiota structure among FAW larvae fed on different host plants. Firmicutes emerged as the dominant phylum, with Enterococcaceae as the dominant family and Enterococcus as the prominent genus. Notably, Enterococcus casseliflavus was frequently observed in the gut microbiota of FAW larvae across host plants. Metabolism pathways, particularly those related to carbohydrate and amino acid metabolism, played a crucial role in the adaptation of the FAW gut microbiota to different host plants. KEGG orthologs associated with the regulation of the peptide/nickel transport system permease protein in sorghum-fed larvae and the 6-phospho-ß-glucosidase gene linked to glycolysis/gluconeogenesis as well as starch and sucrose metabolism in pepper-fed larvae were identified. Moreover, the study identified the top 20 ARGs in the gut microbiota of FAW larvae fed on different host plants, with the maize-fed group exhibiting the highest abundance of vanRC. CONCLUSIONS: Our metagenomic sequencing study reveals significant variations in the gut microbiota composition and function of FAW larvae across diverse host plants. These findings underscore the intricate co-evolutionary relationship between hosts and their gut microbiota, suggesting that host transfer profoundly influences the gut microbiota and, consequently, the adaptability and pest management strategies for FAW.
Assuntos
Bactérias , Microbioma Gastrointestinal , Larva , Metagenômica , Sorghum , Spodoptera , Zea mays , Animais , Spodoptera/microbiologia , Spodoptera/genética , Larva/microbiologia , Microbioma Gastrointestinal/genética , Zea mays/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Sorghum/microbiologia , Solanum lycopersicum/microbiologia , Capsicum/microbiologia , MetagenomaRESUMO
Influenza and Newcastle disease are the most important poultry diseases that cause high annual damage to poultry farms worldwide. Newcastle virus fusion (F) gene and Influenza Virus Hemagglutinin (HA) gene are capable of encoding F and HA proteins that are the main factors in creating immunity, so this study aimed to clone and express these genes in Spodoptera frugiperda (Sf9) cells using baculovirus expression system. After isolating the Newcastle and Influenza virus genome, the HA gene of influenza virus and the F gene of Newcastle virus were amplified by reverse transcriptase PCR and specific primers and then cloned into pFastBacTM Dual plasmid. A recombinant sucker with these genes was produced in the DH10Bac host cell. By transfecting Sf9 cells with recombinant bacmid, expression was assessed by SDS-PAGE, western blotting, and Bradford methods. Cloning of genes into the bacmid was successful. By transfecting the recombinant bacmid into Spodoptera frugiperda cells, 218 µg/ml of the recombinant protein was obtained in the supernatant. In addition, the presence of protein was confirmed by western blotting. The PCR products of HA and F genes showed one band of 1.7 kb size using specific primers. The pFastHA1 vector was about 7 kb in size. Two bands of about 7 kb and 1.7 kb were created by ligation of the F gene and pFastHA1 vector based on enzymatic digestion, indicating the correct ligation of F gene under the P10 promoter. This is the first report on the cloning and Co-expression of two HA and F genes using baculovirus expression system and can be a candidate for dual influenza and Newcastle vaccine. Mixtures of these recombinant proteins can be used as vaccine candidates against both avian influenza and Newcastle disease.
Assuntos
Baculoviridae , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Vírus da Influenza A Subtipo H9N2 , Vírus da Doença de Newcastle , Spodoptera , Animais , Baculoviridae/genética , Células Sf9 , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Vírus da Influenza A Subtipo H9N2/genética , Vírus da Doença de Newcastle/genética , Proteínas Virais de Fusão/genética , Proteínas Virais de Fusão/metabolismo , Expressão Gênica , Clonagem Molecular/métodos , Vetores Genéticos/genéticaRESUMO
MicroRNAs (miRNAs) represent a class of short, non-coding RNAs that are widely acknowledged as crucial participants in virus-host interactions. MiR-184, a highly conserved and abundant miRNA in insects, has yet to be extensively studied for its involvement in baculovirus infection. In this study, we investigated how miR-184 affects the infection and replication of Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The results indicated that after AcMNPV infection, there was an initial increase in the expression of miR-184 within 24 h, followed by a subsequent decrease. MiR-184 can inhibit AcMNPV's DNA replication and budded virus production by directly targeting four viral genes, namely ie1, ac66, p49, and lef9. Moreover, suppressing miR-184 expression enhanced the insecticidal efficacy of AcMNPV against Spodoptera exigua larvae and markedly elevated the host ATPase gene expressions. These findings showed that miR-184 had a substantial impact on the interactions between baculoviruses and insects, presenting a prospective candidate for developing highly effective miRNA-based biopesticides.
Assuntos
MicroRNAs , Nucleopoliedrovírus , Spodoptera , Replicação Viral , Nucleopoliedrovírus/genética , Nucleopoliedrovírus/fisiologia , MicroRNAs/genética , MicroRNAs/metabolismo , Animais , Spodoptera/virologia , Spodoptera/genética , Células Sf9 , Larva/virologia , Larva/genéticaRESUMO
There is a growing interest in the effects of climate warming on olfaction, as temperature may affect this essential sense. In insects, the response of the olfactory system to developmental temperature might be mediated by body size or mass because body size and mass are negatively affected by developmental temperature in most ectotherms. We tested this hypothesis of a mass-mediated effect of developmental temperature on olfaction in the moth Spodoptera littoralis. We measured the olfactory sensitivity of male to female sex pheromone and five plant odors using electroantennography. We compared males reared at an optimal temperature (25 °C with a daily fluctuation of ±5 °C) and at a high temperature (33 ± 5 °C) close to the upper limit of S. littoralis. On average, the olfactory sensitivity of males did not differ between the two developmental temperatures. However, our analyses revealed an interaction between the effects of developmental temperature and body mass on the detection of the six chemicals tested. This interaction is explained by a positive relationship between antennal sensitivity and body mass observed only with the high developmental temperature. Our results show that the effect of developmental temperature may not be detected when organism size is ignored.
Assuntos
Olfato , Spodoptera , Temperatura , Animais , Masculino , Olfato/fisiologia , Feminino , Spodoptera/crescimento & desenvolvimento , Spodoptera/fisiologia , Peso Corporal , Atrativos Sexuais/metabolismo , Odorantes , Tamanho Corporal , Antenas de Artrópodes/fisiologia , Antenas de Artrópodes/crescimento & desenvolvimento , Mariposas/crescimento & desenvolvimento , Mariposas/fisiologiaRESUMO
Cytochrome P450 monooxygenases in insects have been verified to implicated in insecticide and phytochemical detoxification metabolism. However, the regulation of P450s, which are modulated by signal-regulated transcription factors (TFs), is less well studied in insects. Here, we found that the Malpighian tubule specific P450 gene SlCYP9A75b in Spodoptera litura is induced by xenobiotics. The transgenic Drosophila bioassay and RNAi results indicated that this P450 gene contributes to α-cypermethrin, cyantraniliprole, and nicotine tolerance. In addition, functional analysis revealed that the MAPKs p38, PI3K/Akt, and JAK-STAT activate the transcription factor fushi tarazu factor 1 (FTZ-F1) to regulate CYP9A75b expression. These findings provide mechanistic insights into the contributions of CYP9A genes to xenobiotic detoxification and support the possible involvement of different signaling pathways and TFs in tolerance to xenobiotics in insects.
Assuntos
Sistema Enzimático do Citocromo P-450 , Proteínas de Insetos , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais , Spodoptera , Xenobióticos , Animais , Spodoptera/genética , Spodoptera/efeitos dos fármacos , Spodoptera/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Xenobióticos/metabolismo , Xenobióticos/farmacologia , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Inseticidas/farmacologia , Fator de Transcrição STAT5/metabolismo , Fator de Transcrição STAT5/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
The long-term use of agricultural insecticides has led to the development of resistant strains. In this context, the isoxazoline structure has become an active area of pesticide research owing to its wide insecticidal spectrum, nontoxicity to mammals, and lack of cross-resistance with known insecticides. In the present study, based on the discovery of compound G22 in our previous work, a series of novel isoxazoline compounds containing acylhydrazine were designed and synthesized using a scaffold hopping strategy. The insecticidal activities of the target compounds were assessed, and compound L17 (LC50 = 0.489 mg/L) showed insecticidal activity against Spodoptera frugiperda superior to those of the commercial insecticides indoxacarb (LC50 = 3.14 mg/L) and fluralaner (LC50 = 0.659 mg/L). Theoretical calculations indicated that the introduction of acylhydrazine plays an important role in the biological activity of the target compounds. Furthermore, the enzyme-linked immunosorbent assay and molecular docking results indicated that L17 may act on the GABA receptor of the target insect. These results indicated that L17 is a potential candidate compound for controlling S. frugiperda populations in agriculture.
Assuntos
Desenho de Fármacos , Hidrazinas , Inseticidas , Isoxazóis , Simulação de Acoplamento Molecular , Spodoptera , Inseticidas/química , Inseticidas/síntese química , Inseticidas/farmacologia , Animais , Hidrazinas/química , Hidrazinas/síntese química , Hidrazinas/farmacologia , Spodoptera/efeitos dos fármacos , Relação Estrutura-Atividade , Isoxazóis/química , Isoxazóis/farmacologia , Isoxazóis/síntese química , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Estrutura MolecularRESUMO
Spodoptera frugiperda has emerged as a major invasive pest worldwide. The utilization of chemical pesticides not only poses numerous ecological concerns but also fosters resistance in S. frugiperda. In this study, we designed and synthesized three novel thiothiazolidinone compounds (6a, 7b, and 7e) and incorporated innovative thiothiazolidinone structural elements into the piperine skeleton. Treatment with compounds 6a and 7e resulted in the blackening and agglomeration of oviduct eggs within the ovaries of certain female moths, impeding the release of normal eggs. The levels of vitellogenin and vitellogenin receptor, along with three trehalase inhibitors, exhibited a dynamic equilibrium state, leading to no discernible change in egg production but a notable increase in the generation of low-hatching-rate egg fragments. Compared with the injection of 2%DMSO, the eclosion rate of 6a injection was significantly decreased, as followed the spawning time and longevity were prolonged or significantly prolonged in the trehalase inhibitors of 6a, 7b, and 7e. We aimed to investigate the regulatory impacts of three new pepper thiothiazolidinone compounds on the reproduction of S. frugiperda, and to authenticate the efficacy of novel alginase inhibitors in inhibiting the reproduction of S. frugiperda. This research endeavors to aid in the identification of efficient and steadfast trehalase inhibitors, thereby expediting the research and development of potent biological pesticides.
Assuntos
Fertilidade , Spodoptera , Animais , Spodoptera/efeitos dos fármacos , Spodoptera/fisiologia , Feminino , Fertilidade/efeitos dos fármacos , Inseticidas/farmacologia , Capsicum , Trealase/metabolismo , Trealase/antagonistas & inibidores , Vitelogeninas/metabolismo , Tiazolidinas/farmacologiaRESUMO
Arecoline (ACL), an active constituent derived from Areca catechu L., exerts various pharmacological effects and serves as a potential plant-based insecticide. However, the effects of ACL on Spodoptera litura, an important and widely distributed agricultural pest, remain unknown. This study aimed to elucidate the mechanism underlying ACL-induced toxicity and its inhibitory effects on larval growth and development through intestinal pathology observations, intestinal transcriptome sequencing, intestinal digestive enzyme activity analysis. The results indicated that ACL exposure leads to pathological alterations in the S. litura midgut. Furthermore, the detection of digestive enzyme activity revealed that ACL inhibits the activities of acetyl CoA carboxylase, lipase, α-amylase, and trypsin. Simultaneously, upregulation of superoxide dismutase activity and downregulation of malondialdehyde levels were observed after ACL exposure. Transcriptome analysis identified 1118 genes that were significantly differentially expressed in the midgut after ACL exposure, potentially related to ACL toxic effects. Notably, ACL treatment downregulated key enzymes involved in lipid metabolism, such as fatty acid binding protein 2-like, pancreatic triacylglycerol lipase-like, pancreatic lipid-related protein 2-like, and fatty acid binding protein 1-like. Taken together, these results suggest that ACL induces midgut damage and impedes larval growth by suppressing digestive enzyme activity in the intestine. These findings can aid in the development of environmentally friendly plant-derived insecticides, utilizing ACL to effectively combat S. litura proliferation.
Assuntos
Intestinos , Larva , Spodoptera , Animais , Spodoptera/efeitos dos fármacos , Spodoptera/crescimento & desenvolvimento , Larva/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Inseticidas/toxicidade , Inseticidas/farmacologia , Lipase/metabolismo , Lipase/genéticaRESUMO
Heat shock proteins (Hsps) are stress response proteins. In a previous study, host larval Hsp70s were identified as the structural proteins of virions of Heliothis virescens ascovirus 3h (HvAV-3h), an insect virus that mainly infects noctuid larvae. To investigate the response of hsp70s of healthy Mythimna separata, Spodoptera exigua, Spodoptera frugiperda, and Spodoptera litura larvae to various abiotic or entomopathogenic stresses, quantitative PCR was used to detect larval hsp70s expression patterns. Results showed distinct expression patterns of hsp70s in response to different abiotic stresses. Notably, Mshsp70 expression pattern resembled Slhsp70 under most treatments. In healthy larvae, no tissue tropism was observed concerning the relative expression of Mshsp70, Sfhsp70, and Slhsp70. After infection with HvAV-3h, the expression of hsp70s in all dissected tissues of all tested larval species increased. Significant differences were found in the fat bodies of M. separata, S. exigua, and S. litura as well as in the hemolymph of S. exigua and S. litura. Subsequent silencing of Slhsp70, resulted in a significant decrease in DNA replication levels of HvAV-3h in S. litura larvae at 24 and 72 h post RNA interference, indicating that Slhsp70 is necessary for DNA replication in HvAV-3h. These data can provide references for the studying on the stress response of noctuid larvae to different environmental factors.
Assuntos
Proteínas de Choque Térmico HSP70 , Larva , Estresse Fisiológico , Animais , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Larva/genética , Larva/metabolismo , Estresse Fisiológico/genética , Spodoptera/genética , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Mariposas/genética , Ascoviridae/genética , Ascoviridae/metabolismoRESUMO
Insect growth-blocking peptides (GBPs) are a family of cytokines found in several insect orders and are known for their roles in regulating development, paralysis, cell proliferation, and immune responses. Despite their diverse functions, the potential of GBPs as biocontrol targets against the pest Spodoptera frugiperda (Lepidoptera: Noctuidae) has not been fully explored. In this study, S. frugiperda GBP (SfGBP) was identified and functionally characterized. SfGBP is synthesized as a 146 amino acid proprotein with a 24 amino acid C-terminal active peptide (Glu123-Gly146). Predominant expression of SfGBP occurs in fourth to sixth instar larvae and in the larval fat body, with significant upregulation in response to pathogens and pathogen-associated molecular patterns. Injection of the synthetic active peptide into larvae induced growth retardation, delayed pupation, and increased survival against Beauveria bassiana infection. Conversely, RNA interference-mediated knockdown of SfGBP resulted in accelerated growth, earlier pupation, and decreased survival against B. bassiana infection. Further analysis revealed that SfGBP promoted SF9 cell proliferation and spreading, enhanced bacteriostatic activity of larval hemolymph, and directly inhibited germination of B. bassiana conidia. In addition, SfGBP enhanced humoral responses, such as upregulation of immunity-related genes and generation of reactive oxygen species, and cellular responses, such as nodulation, phagocytosis, and encapsulation. These results highlight the dual regulatory role of SfGBP in development and immune responses and establish it as a promising biocontrol target for the management of S. frugiperda.
Assuntos
Proteínas de Insetos , Larva , Spodoptera , Animais , Spodoptera/efeitos dos fármacos , Spodoptera/imunologia , Proteínas de Insetos/metabolismo , Proteínas de Insetos/genética , Larva/efeitos dos fármacos , Larva/imunologia , Beauveria/fisiologia , Sequência de Aminoácidos , Controle Biológico de Vetores/métodosRESUMO
The fall armyworm (Spodoptera frugiperda) poses a substantial threat to many important crops worldwide, emphasizing the need to develop and implement advanced technologies for effective pest control. CRISPR/Cas9, derived from the bacterial adaptive immune system, is a prominent tool used for genome editing in living organisms. Due to its high specificity and adaptability, the CRISPR/Cas9 system has been used in various functional gene studies through gene knockout and applied in research to engineer phenotypes that may cause economical losses. The practical application of CRISPR/Cas9 in diverse insect orders has also provided opportunities for developing strategies for genetic pest control, such as gene drive and the precision-guided sterile insect technique (pgSIT). In this review, a comprehensive overview of the recent progress in the application of the CRISPR/Cas9 system for functional gene studies in S. frugiperda is presented. We outline the fundamental principles of applying CRISPR/Cas9 in S. frugiperda through embryonic microinjection and highlight the application of CRISPR/Cas9 in the study of genes associated with diverse biological aspects, including body color, insecticide resistance, olfactory behavior, sex determination, development, and RNAi. The ability of CRISPR/Cas9 technology to induce sterility, disrupt developmental stages, and influence mating behaviors illustrates its comprehensive roles in pest management strategies. Furthermore, this review addresses the limitations of the CRISPR/Cas9 system in studying gene function in S. frugiperda and explores its future potential as a promising tool for controlling this insect pest.
Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Spodoptera , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Animais , Spodoptera/genéticaRESUMO
Fall armyworm (FAW), Spodoptera frugiperda is a generalist pest known to feed on more than 300 plant species, including major staple crops such as rice, maize and sorghum. Biological control of FAW using a combination of a major indigenous egg parasitoid Telenomus remus and entomopathogenic fungi was explored in this study. Metarhizium anisopliae strains (ICIPE 7, ICIPE 41, and ICIPE 78) and Beauveria bassiana ICIPE 621 which demonstrated effectiveness to combat the pest, were evaluated through direct and indirect fungal infection to assess their pathogenicity and virulence against T. remus adults, S. frugiperda eggs and their effects on T. remus parasitism rates. Metarhizium anisopliae ICIPE 7 and ICIPE 78 exhibited the highest virulence against T. remus adults with LT50 values >2 days. ICIPE 7 induced the highest T. remus mortality rate (81.40 ± 4.17%) following direct infection with dry conidia. Direct fungal infection also had a significant impact on parasitoid emergence, with the highest emergence rate recorded in the M. anisopliae ICIPE 7 treatment (42.50 ± 5.55%), compared to the control ± (83.25 ± 5.94%). In the indirect infection, the highest concentration of 1 x 109 conidia ml-1 of ICIPE 78 induced the highest mortality (100 ± 0.00%) of T. remus adults, and the highest mortality (51.25%) of FAW eggs, whereas the least FAW egg mortality (15.25%) was recorded in the lowest concentration 1 x 105 conidia ml-1 of ICIPE 41. The number of parasitoids that emerged and their sex ratios were not affected by the different fungal strain concentrations except in ICIPE 7 at high dose. This study showed that potential combination of both M. anisopliae and B. bassiana with T. remus parasitoid can effectively suppress FAW populations.
Assuntos
Beauveria , Metarhizium , Controle Biológico de Vetores , Spodoptera , Animais , Beauveria/patogenicidade , Beauveria/isolamento & purificação , Controle Biológico de Vetores/métodos , Metarhizium/patogenicidade , Spodoptera/microbiologia , Spodoptera/parasitologia , Virulência , Feminino , Vespas/microbiologia , Heterópteros/microbiologia , Heterópteros/parasitologia , Óvulo/microbiologia , Agentes de Controle Biológico , Masculino , Análise de SobrevidaRESUMO
BACKGROUND: Normalization with respect to stable housekeeping genes is important to facilitate gene transcription regulation research and acquire more accurate quantitative polymerase chain reaction (qPCR) data. In the current study, five candidates housekeeping genes of the cotton leafworm, Spodoptera littoralis encoding for Actin (Actin), elongation factor 1-alpha (EF1α), ribosomal protein S3 (RPS3), ribosomal protein 49 (RP49), and Ubiquitin (Ubi), were evaluated as normalization housekeeping genes under Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) viral infection. METHODS AND RESULTS: The qPCR results confirmed the expression of all five housekeeping genes in S. littoralis viral infected larvae. The expression profiles of the housekeeping genes showed that the EF1α, Actin, and RP49 had the minimum average Ct values of 18.41 ± 0.66, 18.84 ± 0.90 and 19.01 ± 0.87 in all infected samples, respectively. While RPS3 and Ubi showed the maximum average Ct of 21.61 ± 0.51 and 21.11 ± 0.82, respectively. According to the results of ΔCt and geNorm analysis, EF1α was ranked as the most stable housekeeping gene during infection time-course. While by using BestKeeper, geNorm and NormFinder, the Ubi, RP49, and RPS3 showed the most genes transcription stability. The obtained results were also validated using the Cytochrome c oxidase (COX) gene transcripts in response to SpliNPV infection. CONCLUSIONS: The results revealed that EF1α and Ubi were the most stable housekeeping genes to be used for normalizing S. littoralis gene transcription regulation under SpliNPV infection. These findings, provide a significant addition for gene transcription regulation studies of S. littoralis upon infection using SpliNPV as a bio-agent.
Assuntos
Genes Essenciais , Nucleopoliedrovírus , Spodoptera , Animais , Spodoptera/genética , Spodoptera/virologia , Genes Essenciais/genética , Nucleopoliedrovírus/genética , Regulação da Expressão Gênica , Larva/genética , Larva/virologia , Transcrição Gênica/genética , Perfilação da Expressão Gênica/métodos , Proteínas de Insetos/genéticaRESUMO
Trichomes are specialized epidermal outgrowths covering the aerial parts of most terrestrial plants. There is a large species variability in occurrence of different types of trichomes such that the molecular regulatory mechanism underlying the formation and the biological function of trichomes in most plant species remain unexplored. Here, we used Chrysanthemum morifolium as a model plant to explore the regulatory network in trichome formation and terpenoid synthesis and unravel the physical and chemical roles of trichomes in constitutive defense against herbivore feeding. By analyzing the trichome-related genes from transcriptome database of the trichomes-removed leaves and intact leaves, we identified CmMYC2 to positively regulate both development of T-shaped and glandular trichomes as well as the content of terpenoids stored in glandular trichomes. Furthermore, we found that the role of CmMYC2 in trichome formation and terpene synthesis was mediated by interaction with CmMYBML1. Our results reveal a sophisticated molecular mechanism wherein the CmMYC2-CmMYBML1 feedback inhibition loop regulates the formation of trichomes (non-glandular and glandular) and terpene biosynthesis, collectively contributing to the enhanced resistance to Spodoptera litura larvae feeding. Our findings provide new insights into the novel regulatory network by which the plant synchronously regulates trichome density for the physical and chemical defense against herbivory.
Assuntos
Chrysanthemum , Regulação da Expressão Gênica de Plantas , Herbivoria , Proteínas de Plantas , Terpenos , Tricomas , Tricomas/metabolismo , Terpenos/metabolismo , Chrysanthemum/genética , Chrysanthemum/metabolismo , Chrysanthemum/fisiologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Animais , Defesa das Plantas contra Herbivoria , Folhas de Planta/metabolismo , Genes de Plantas , Spodoptera/fisiologiaRESUMO
Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this goal. Unfortunately, the adaptability of insects has led to resistance to certain proteins in current commercial products. Therefore, new insecticidal traits representing a different mode of action (MoA) than those currently in use are needed. Cry1Ja has good insecticidal activity against various lepidopteran species, and it provides robust protection against insect feeding with in planta expression. For Bt proteins, different MoAs are determined by their binding sites in the insect midgut. In this study, competitive binding assays are performed using brush border membrane vesicles (BBMVs) from Helicoverpa zea, Spodoptera frugiperda, and Chrysodeixis includens to evaluate the MoA of Cry1Ja relative to representatives of the various Bt proteins that are expressed in current commercial products for lepidopteran insect protection. This study highlights differences in the shared Cry protein binding sites in three insect species, Cry1Ja bioactivity against Cry1Fa resistant FAW, and in planta efficacy against target pests. These data illustrate the potential of Cry1Ja for new insect trait development.
Assuntos
Toxinas de Bacillus thuringiensis , Proteínas de Bactérias , Endotoxinas , Proteínas Hemolisinas , Controle Biológico de Vetores , Animais , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/farmacologia , Toxinas de Bacillus thuringiensis/metabolismo , Endotoxinas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Inseticidas/farmacologia , Spodoptera/efeitos dos fármacos , Microvilosidades/metabolismo , Microvilosidades/efeitos dos fármacos , Controle de Insetos/métodos , Bacillus thuringiensis/genética , Plantas Geneticamente Modificadas/genéticaRESUMO
Exploring the mechanism of microbiota assembly and its ecological consequences is crucial for connecting microbiome variation to ecosystem function. However, the influencing factors underlying microbiota assembly in the host-microbe system and their impact on the host phenotype remain unclear. Through investigating the prevalent and worsening ecological phenomenon of insecticide resistance in global agriculture, we found that insecticide exposure significantly changed the gut microbiota assembly patterns of a major agricultural invasive insect pest, Spodoptera frugiperda. The relative importance of various microbiota assembly processes significantly varied with habitat heterogeneity and heterogeneous selection serving as a potential predictor of the host's insecticide resistance in field populations. Moreover, disturbance of the gut microbiota assembly through antibiotics was revealed to significantly affect the rate and heritability of insecticide resistance evolution, leading to a delay in insecticide resistance evolution in this insect pest. These findings indicate that the gut microbiota assembly process of the insect host is influenced by persistent exposure to habitat conditions, particularly insecticides. This variation in insecticide exposure-related community assembly process subsequently influences the insect host's insecticide resistance phenotype. This study provides insights into gut microbiota assembly processes from a symbiotic perspective and underscores the significant impact of symbiotic community changes on host phenotypic variation.
Assuntos
Microbioma Gastrointestinal , Resistência a Inseticidas , Inseticidas , Spodoptera , Simbiose , Animais , Spodoptera/microbiologia , Spodoptera/efeitos dos fármacos , Spodoptera/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacosRESUMO
MicroRNAs (miRNAs) are endogenous, non-coding RNAs, which are functional in a variety of biological processes through post-transcriptional regulation of gene expression. However, the role of miRNAs in the interaction between Bacillus thuringiensis and insects remains unclear. In this study, small RNA libraries were constructed for B. thuringiensis-infected (Bt) and uninfected (CK) Spodoptera exigua larvae (treated with double-distilled water) using Illumina sequencing. Utilising the miRDeep2 and Randfold, a total of 233 known and 726 novel miRNAs were identified, among which 16 up-regulated and 34 down-regulated differentially expressed (DE) miRNAs were identified compared to the CK. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that potential target genes of DE miRNAs were associated with ABC transporters, fatty acid metabolism and MAPK signalling pathway which are related to the development, reproduction and immunity. Moreover, two miRNA core genes, SeDicer1 and SeAgo1 were identified. The phylogenetic tree showed that lepidopteran Dicer1 clustered into one branch, with SeDicer1 in the position closest to Spodoptera litura Dicer1. A similar phylogenetic relationship was observed in the Ago1 protein. Expression of SeDicer1 increased at 72 h post infection (hpi) with B. thuringiensis; however, expression of SeDicer1 and SeAgo1 decreased at 96 hpi. The RNAi results showed that the knockdown of SeDicer1 directly caused the down-regulation of miRNAs and promoted the mortality of S. exigua infected by B. thuringiensis GS57. In conclusion, our study is crucial to understand the relationship between miRNAs and various biological processes caused by B. thuringiensis infection, and develop an integrated pest management strategy for S. exigua via miRNAs.
Assuntos
Bacillus thuringiensis , MicroRNAs , Spodoptera , Animais , Spodoptera/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Bacillus thuringiensis/genética , Larva/genética , Perfilação da Expressão GênicaRESUMO
Plant-soil interactions have bottom-up and top-down effects within a plant community. Heavy metal pollution can change plant-soil interactions, directly influence bottom-up effects and indirectly affect herbivores within the community. In turn, herbivores can affect plant-soil interactions through top-down effects. However, the combined effects of heavy metals and herbivores on soil enzymes, plants and herbivores have rarely been reported. Therefore, the effects of lead (Pb), Spodoptera litura and their combined effects on soil enzyme activities, pakchoi nutrition, defence compounds and S. litura fitness were examined here. Results showed that Pb, S. litura and their combined effects significantly affected soil enzymes, pakchoi and S. litura. Specifically, exposure to double stress (Pb and S. litura) decreased soil urease, phosphatase and sucrase activities compared with controls. Furthermore, the soluble protein and sugar contents of pakchoi decreased, and the trypsin inhibitor content and antioxidant enzyme activity increased. Finally, the S. litura development period was extended, and survival, emergence rates and body weight decreased after exposure to double stress. The combined stress of Pb and S. litura significantly decreased soil enzyme activities. Heavy metal accumulation in plants may create a superposition or synergistic effect with heavy metal-mediated plant chemical defence, further suppressing herbivore development. Pb, S. litura and their combined effects inhibited soil enzyme activities, improved pakchoi resistance and reduced S. litura development. The results reveal details of soil-plant-herbivore interactions and provide a reference for crop pest control management in the presence of heavy metal pollution.
Assuntos
Chumbo , Solo , Spodoptera , Animais , Spodoptera/efeitos dos fármacos , Spodoptera/crescimento & desenvolvimento , Spodoptera/enzimologia , Chumbo/toxicidade , Solo/química , Herbivoria , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/enzimologiaRESUMO
Spodoptera litura is a significant agricultural pest, and its glutathione S-transferase (GST) plays a crucial role in insecticide resistance. This study aimed to investigate the relationship between the SlGSTe11 gene of S. litura and resistance to cyantraniliprole and nicotine. Transcriptome analysis revealed that SlGSTe11 is highly expressed mainly in fat bodies, with a significant increase in SlGSTe11 gene expression under induction by cyantraniliprole and nicotine. The ectopic expression of the SlGSTe11 gene in transgenic fruit flies resulted in a 5.22-fold increase in the tolerance to cyantraniliprole. Moreover, compared to the UAS-SlGSTe11 line, the Act5C-UAS>SlGSTe11 line laid more eggs and had a lower mortality after nicotine exposure. RNAi-mediated inhibition of SlGSTe11 gene expression led to a significant increase in the mortality of S. litura under cyantraniliprole exposure. In vitro metabolism experiments demonstrated that the recombinant SlGSTe11 protein efficiently metabolizes cyantraniliprole. Molecular docking results indicated that SlGSTe11 has a strong affinity for both cyantraniliprole and nicotine. These findings suggest that SlGSTe11 is involved in the development of resistance to cyantraniliprole and nicotine in S. litura.