Transcriptomics and interactomics during the primary infection of an SfNPV baculovirus on Spodoptera frugiperda larvae
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ABSTRACT:
SUBMITTER: Rangel-Nunez J
PROVIDER: S-EPMC10703053 | biostudies-literature | 2023 Jan
REPOSITORIES: biostudies-literature
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Project description:Baculovirus infection of Spodoptera frugiperda cells is a system of choice to produce a range of recombinant proteins, vaccines and, potentially, gene therapy vectors. While baculovirus genomes are well characterized, the genome of S. frugiperda is not sequenced and the virus-host molecular interplay is sparsely known. Herein, we describe the application of stable isotope labeling by amino acids in cell culture (SILAC) to obtain the first comparative proteome quantitation of S. frugiperda cells during growth and early baculovirus infection. The proteome coverage was maximized by compiling a search database with protein annotations from insect species. Of interest were differentially proteins related to energy metabolism, endoplasmic reticulum and oxidative stress, yet not investigated in the scope of baculovirus infection. Further, the reduced expression of key viral-encoded proteins early in the infection cycle is suggested to be related with decreased viral replication at high cell density culture. These findings have implications for virological research and improvement of baculovirus-based bioprocesses.
| S-EPMC3196586 | biostudies-literature
Project description:MicroRNAs (miRNAs) as small non-coding RNAs play important roles in many biological processes such as development, cell signalling and immune response. Studies also suggest that miRNAs are important in host–virus interactions where the host limits virus infection by differentially expressing miRNAs that target essential viral genes. Here, we identified conserved and new miRNAs from Spodoptera frugiperda cells (Sf9) using a combination of deep sequencing and bioinformatics as well as experimental approaches. S. frugiperda miRNAs share common features of miRNAs in other organisms, such as uracil (U) at the 59 end of miRNA. The 59 ends of the miRNAs were more conserved than the 39 ends, revealing evolutionary protection of the seed region in miRNAs. The predominant miRNAs were found to be conserved among arthropods. The majority of homologous miRNAs were found in Bombyx mori, with 76 of the 90 identified miRNAs. We found that seed shifting and arm switching have happened in this insect’s miRNAs. Expression levels of the majority of miRNAs changed following baculovirus infection. Results revealed that baculovirus infection mainly led to an overall suppression of cellular miRNAs. We found four different genes being regulated by sfr-miR-184 at the post-transcriptional level. The data presented here further support conservation of miRNAs in insects and other organisms. In addition, the results reveal a differential expression of host miRNAs upon baculovirus infection, suggesting their potential roles in host–virus interactions. Seed shifting and arm switching happened during evolution of miRNAs in different insects and caused miRNA diversification, which led to changes in the target repository of miRNAs.
2015-06-11 | GSE60064 | GEO
Project description:MicroRNAs (miRNAs) as small non-coding RNAs play important roles in many biological processes such as development, cell signalling and immune response. Studies also suggest that miRNAs are important in host–virus interactions where the host limits virus infection by differentially expressing miRNAs that target essential viral genes. Here, we identified conserved and new miRNAs from Spodoptera frugiperda cells (Sf9) using a combination of deep sequencing and bioinformatics as well as experimental approaches. S. frugiperda miRNAs share common features of miRNAs in other organisms, such as uracil (U) at the 59 end of miRNA. The 59 ends of the miRNAs were more conserved than the 39 ends, revealing evolutionary protection of the seed region in miRNAs. The predominant miRNAs were found to be conserved among arthropods. The majority of homologous miRNAs were found in Bombyx mori, with 76 of the 90 identified miRNAs. We found that seed shifting and arm switching have happened in this insect’s miRNAs. Expression levels of the majority of miRNAs changed following baculovirus infection. Results revealed that baculovirus infection mainly led to an overall suppression of cellular miRNAs. We found four different genes being regulated by sfr-miR-184 at the post-transcriptional level. The data presented here further support conservation of miRNAs in insects and other organisms. In addition, the results reveal a differential expression of host miRNAs upon baculovirus infection, suggesting their potential roles in host–virus interactions. Seed shifting and arm switching happened during evolution of miRNAs in different insects and caused miRNA diversification, which led to changes in the target repository of miRNAs. Identification of miRNA and other small non coding RNA in NPV infected Sf9 cells
2015-06-11 | E-GEOD-60064 | biostudies-arrayexpress
Project description:Ascoviruses are large, enveloped DNA viruses that induce remarkable changes in cellular architecture during which the cell is partitioned into numerous vesicles for viral replication. Previous studies have shown that these vesicles arise from a process resembling apoptosis yet which differs after nuclear lysis in that mitochondria are not degraded but are modified by the virus, changing in size, shape, and motility. Moreover, infection does not provoke an obvious innate immune response. Thus, we used in vivo RNA sequencing to determine whether infection by the Spodoptera frugiperda ascovirus 1a (SfAV-1a) modified expression of host mitochondrial, cytoskeletal, and innate immunity genes. We show that transcripts from many mitochondrial genes were similar to those from uninfected controls, whereas others increased slightly during vesicle formation, including those for ATP6, ATP8 synthase, and NADH dehydrogenase subunits, supporting electron microscopy (EM) data that these organelles were conserved for virus replication. Transcripts from 58 of 106 cytoskeletal genes studied increased or decreased more than 2-fold postinfection. More than half coded for mitochondrial motor proteins. Similar increases occurred for innate immunity transcripts and their negative regulators, including those for Toll, melanization, and phagocytosis pathways. However, those for many antimicrobial peptides, such as moricin, increased more than 20-fold. In addition, transcripts for gloverin-3, spod_x_tox, Hdd23, and lebocin, also antimicrobial, increased more than 20-fold. Interestingly, a phenoloxidase inhibitor transcript increased 12-fold, apparently to interfere with melanization. SfAV-1a destroys most fat body cells by 7 days postinfection, so innate immunity gene transcripts apparently occur in remaining cells in this tissue and possibly other major tissues, namely, epidermis and tracheal matrix.IMPORTANCE Ascoviruses are large DNA viruses that infect insects, inducing a cellular pathology that resembles apoptosis but which differs by causing enormous cellular hypertrophy followed by cleavage of the cell into numerous viral vesicles for replication. Previous EM studies suggest that mitochondria are important for vesicle formation. Transcriptome analyses of Spodoptera frugiperda larvae infected with SfAV-1a showed that mitochondrial transcripts were similar to those from uninfected controls or increased slightly during vesicle formation, especially for ATP6, ATP8 synthase, and NADH dehydrogenase subunits. This pattern resembles that for chronic disease-inducing viruses, which conserve mitochondria, differing markedly from viruses causing short-term viral diseases, which degrade mitochondrial DNA. Though mitochondrial transcript increases were low, our results demonstrate that SfAV-1a alters host mitochondrial expression more than any other virus. Regarding innate immunity, although SfAV-1a destroys most fat body cells, certain immunity genes were highly upregulated (greater than 20-fold), suggesting that these transcripts may originate from other tissues.
| S-EPMC7163118 | biostudies-literature
Project description:The fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae) is a recent invasive pest species that has successfully established across sub-Saharan Africa where it continues to disrupt agriculture, particularly smallholder cereal production. Management of FAW in its native range in the Americas has led to the development of resistance to many commercial pesticides before its arrival in Africa. Pesticide use may therefore be ineffective for FAW control in Africa, so new and more sustainable approaches to pest management are required that can help reduce the impact of this exotic pest. Pesticidal plants provide an effective and established approach to pest management in African smallholder farming and recent research has shown that their use can be cost-beneficial and sustainable. In order to optimize the use of botanical extracts for FAW control, we initially screened ten commonly used plant species. In laboratory trials, contact toxicity and feeding bioassays showed differential effects. Some plant species had little to no effect when compared to untreated controls; thus, only the five most promising plant species were selected for more detailed study. In contact toxicity tests, the highest larval mortality was obtained from Nicotiana tabacum (66%) and Lippia javanica (66%). Similarly, in a feeding bioassay L. javanica (62%) and N. tabacum (60%) exhibited high larval mortality at the highest concentration evaluated (10% w/v). Feeding deterrence was evaluated using glass-fibre discs treated with plant extracts, which showed that Cymbopogon citratus (36%) and Azadirachta indica (20%) were the most potent feeding deterrents among the pesticidal plants evaluated. In a screenhouse experiment where living maize plants infested with fall armyworm larvae were treated with plant extracts, N. tabacum and L. javanica were the most potent species at reducing foliar damage compared to the untreated control whilst the synthetic pesticide chlorpyrifos was the most effective in reducing fall armyworm foliar damage. Further field trial evaluation is recommended, particularly involving smallholder maize fields to assess effectiveness across a range of contexts.
| S-EPMC7020173 | biostudies-literature
Project description:The insecticidal Vip3 proteins, secreted by Bacillus thuringiensis (Bt) during its vegetative growth phase, are currently used in Bt crops to control insect pests, and are genetically distinct from known insecticidal Cry proteins. Compared with Cry toxins, the mechanisms of Vip3 toxins are still poorly understood. Here, the responses of Spodoptera frugiperda larvae after Vip3Aa challenge are characterized. Using an integrative analysis of transcriptomics and proteomics, we found that Vip3Aa has enormous implications for various pathways. The downregulated genes and proteins were mainly enriched in metabolic pathways, including the insect hormone synthesis pathway, whereas the upregulated genes and proteins were mainly involved in the caspase-mediated apoptosis pathway, along with the MAPK signaling and endocytosis pathways. Moreover, we also identified some important candidate genes involved in apoptosis and MAPKs. The present study shows that exposure of S. frugiperda larvae to Vip3Aa activates apoptosis pathways, leading to cell death. The results will promote our understanding of the host response process to the Vip3Aa, and help us to better understand the mode of action of Vip3A toxins.
| S-EPMC8781260 | biostudies-literature
Project description:The innate immunity of insects encompasses cellular and humoral defense mechanisms and constitutes the primary defense against invading microbial pathogens. Cellular immunity (phagocytosis, nodulation, and encapsulation) is primarily mediated by hemocytes. Plasmatocytes and granulocytes play an important role and require changes in the cytoskeletons of hemocytes. However, research investigating the immunological impacts of insecticides on the fall armyworm (FAW), Spodoptera frugiperda, remains scarce. Therefore, we conducted a study to investigate the effects of chlorantraniliprole exposure on cellular immunity in FAW larvae. Our findings revealed the presence of five types of hemocytes in the larvae: prohemocytes, plasmatocytes, granulocytes, oenocytoids, and spherulocytes. The LD10, LD20, and LD30 of chlorantraniliprole affected both the morphology and total count of some hemocytes in the larvae. Moreover, larvae exposed to chlorantraniliprole showed increased phagocytosis, nodulation, and encapsulation. To determine the mechanism of the enhanced cellular immunity, we studied plasmatocytes in the spread state and the cytoskeleton in hemocytes. It was found that the spreading ratio of plasmatocytes and the areas of the cytoskeletons in hemocytes were increased after chlorantraniliprole treatment. These results suggest that exposure to chlorantraniliprole results in an enhanced immune response function in FAW larvae, which may be mediated by cytoskeletal changes and plasmatocyte spreading. Consequently, this study provides valuable insights into the cellular immune response of FAW larvae to insecticide exposure.
| S-EPMC11354488 | biostudies-literature
Project description:BackgroundSpodoptera litura is an important polyphagous pest that causes significant damage to the agricultural sector. We performed RNA-seq of 15 S. litura individuals from larval (fifth and sixth instar larvae), chrysalis, and adult developmental stages. We also compared the S. litura transcriptome data with Spodoptera frugiperda across the same developmental stages, which was sequenced in our previous study.ResultsA total of 101,885 differentially expressed transcripts (DETs) were identified in S. litura. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that S. litura may undergo active xenobiotic and detoxifying metabolism during its larval and adult stages, which may explain difficulties with current population control measures. We also found that DETs of single-copy orthologous genes between S. litura and S. frugiperda were involved in basic metabolism and development. However, energy and metabolic processes genes had a higher expression in S. litura, whereas nervous and olfactory function genes had a higher expression in S. frugiperda. Metagenomics analysis in larval S. litura and S. frugiperda revealed that microbiota participate in the detoxification and metabolism processes, but the relative abundance of detoxification-related microbiota was more abundant in S. frugiperda. Transcriptome results also confirmed the detoxification-related pathway of S. frugiperda was more abundant than in S. litura.ConclusionsSignificant changes at transcriptional level were identified during the different development stages of S. litura. Importantly, we also identified detoxification associated genes and gut microbiota between S. litura and S. frugiperda at different developmental stages, which will be valuable in revealing possible mechanisms of detoxification and development in these two lepidopterans.
| S-EPMC9123734 | biostudies-literature
Project description:Accurate chromosome segregation requires assembly of the multiprotein kinetochore complex at centromeres. In most eukaryotes, kinetochore assembly is primed by the histone H3 variant CenH3, which physically interacts with components of the inner kinetochore constitutive-centromere-associated-network (CCAN). Unexpected to its critical function, previous work identified that select eukaryotic lineages, including several insects, have lost CenH3, while having retained homologs of the CCAN. These findings imply alternative CCAN assembly pathways in these organisms that function in CenH3-independent manners. Here, we study the composition and assembly of CenH3-independent kinetochores of Lepidoptera. We show that lepidopteran kinetochores consist of previously identified CCAN homologs as well as additional components including a divergent CENP-T homolog, which are required for accurate mitotic progression. Our study focuses on CENP-T that we find both necessary and sufficient to recruit the Mis12 outer kinetochore complex. In addition, CRISPR-mediated gene editing in Bombyx mori establishes an essential function of CENP-T in vivo. Finally, the retention of CENP-T homologs in other independently-derived CenH3-deficient insects indicates a conserved mechanism of kinetochore assembly between these lineages. Our study provides the first functional insights into CCAN-based kinetochore assembly pathways that function independently of CenH3, thus contributing to the emerging picture of an unexpected plasticity to build a kinetochore.
2020-02-18 | PXD016092 | Pride
Project description:Biological control is one of the best options for the sustainable management of the invasive maize pest Spodoptera frugiperda in Africa. However, there is limited knowledge of the efficacy of native natural enemies of S. frugiperda and their potential use in integrated pest management. The endoparasitoid wasp Coccygidium luteum is one of the natural enemies of S. frugiperda in Africa. This study assessed, under laboratory conditions, the effect of C. luteum on the leaf consumption rate of its host. Fifty first instar S. frugiperda larvae were exposed to C. luteum for oviposition and the maize leaf consumption rate of parasitized larvae was assessed and compared to 50 unparasitized larvae from the same cohort. Coccygidiumluteum completed a generation, from egg to adult emergence, in 16.7 days. The leaf consumption rate of parasitized S. frugiperda larvae declined gradually compared to unparasitized larvae and the overall consumption reduction by parasitized S. frugiperda larvae was 89%. Our findings show that C. luteum could reduce damage caused by S. frugiperda to maize farms but, prior to its use in biological control programmes, further studies are needed to assess potential parasitism rates in field conditions and develop a cost-effective mass production system.
| S-EPMC6920753 | biostudies-literature