Insect Science最新文献

The regulation and maintenance of immune homeostasis are essential for animal survival, but the molecular mechanisms are not fully understood. Here, we used the model organism Drosophila melanogaster to uncover a potential mechanism by which the nuclear factor-κB transcription factor Relish and miR-100 cooperatively regulate innate immune homeostasis. We first demonstrated in vitro and in vivo that miR-100 can negatively regulate the immune responses of the Imd pathway by inhibiting the expression of TAK1-associated binding protein 2 (Tab2) gene. Second, we found that Relish, an important transcription factor in the Drosophila Imd pathway, could not only modulate the expressions of antimicrobial peptides (AMPs) to promote immune responses, but also bind to the promoter region of miR-100 and activate its transcription to inhibit immune responses. Third, the dynamic expression of genes profiling indicated that the Relish/miR-100/Tab2 regulatory axis could contribute to innate immune homeostasis in Drosophila. Together, our findings reveal the dual role of Relish in immune regulation, that is, Relish promotes the expression of AMPs to resist pathogen infection in the early immune response, while in the late immune stages, Relish readjusts the expression of miR-100 to negatively control immune responses to avoid excessive immunity thus maintaining immunohomeostasis. Meanwhile, our study provides a new perspective for further understanding the complex regulatory mechanism of immune homeostasis in animals.

Juvenile hormone (JH) plays a pivotal role in regulating post-emergence development and metabolism in previtellogenic female Aedes aegypti mosquitoes. In contrast, yolk protein precursor production and egg maturation after a blood meal are regulated by the steroid hormone 20-hydroxyecdysone, the insulin-like growth factor (IGF)/insulin signaling (IIS) pathway, and the mammalian target of rapamycin (mTOR) pathway. The role of IIS/mTOR signaling in female adults prior to blood feeding has not been thoroughly investigated. In this study, we identified a significant increase in the phosphorylation of key effector proteins in the IIS/mTOR signaling pathway, including eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), ribosomal protein S6 kinase (S6K) and forkhead box protein O1 (FoxO1), in previtellogenic females. In vitro fat body culture experiments suggest that JH induces these phosphorylations through rapid nongenomic signaling mediated by the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mTOR network. RNA interference experiments demonstrated that activation of IIS/mTOR signaling in previtellogenic females modulate metabolic gene expression, promoting the accumulation of energy reserves (glycogen and triglycerides), which influence mosquito fecundity. Additionally, depletion of either the insulin receptor (InR) or the JH receptor Methoprene-tolerant (Met) in adult mosquitoes abolished the phosphorylation of these proteins, indicating that both receptors are involved in JH-induced membrane-initiated signal transduction. Although the precise mechanisms remain unclear, this study uncovers a novel function of the IIS/mTOR pathway in adult mosquitoes before blood feeding, as well as a new mode of JH action through its crosstalk with the IIS pathway.

Due to the rise in global temperatures with climate change, insects, as ectotherms, critically depend on their heat tolerance for survival and reproduction. Heat shock proteins (HSPs) are essential for heat tolerance by averting protein denaturation; however, whether HSPs contribute to reproduction-related heat tolerance remains largely unexplored. The study investigated the reproductive heat tolerance and recovery of Monochamus alternatus, a major forestry pest, in response to heat stress. Alongside impairing the development and viability of reproductive organs and sperm, heat stress was also found to reduce fecundity, fertility, mating, and oviposition behaviors. Remarkably, all reproductive parameters of M. alternatus recovered within 4 weeks postexposure. To investigate the recovery mechanisms, we identified 10 reproduction-related proteins as candidate substrate proteins of an HSP protein in M. alternatus using immunoprecipitation coupled with mass spectrometry analysis. Heat stress inhibited the transcription of these reproduction-related genes, thereby adversely affecting reproductive parameters. However, the induction of HSP20s transcription in response to heat stress appeared to facilitate the refolding of these critical reproduction-related proteins during the recovery phase, thus preventing lasting reproductive damage. Overall, this study suggests that while M. alternatus populations might be vulnerable to climate-induced temperature increases, their fertility can recover, mediated by the interaction of HSPs with reproduction-related genes. These findings offer profound insights into insect heat tolerance and recovery, expanding our understanding of HSP20 proteins' biological functions.

Ultraviolet (UV) radiation, an environmental stressor, is crucial for the survival and adaptation of organisms. Myzus persicae, a global pest, is exposed to sunlight year-round, making it unable to avoid UV rays in its environment. MicroRNAs (miRNAs) are important posttranscriptional regulators of gene expression and mediate various biological processes. However, the role of miRNA in aphids in response to UV-B stress is unclear. In this study, Mpp53 expression level significantly increased with an increase in the duration of UV-B radiation, peaking at 2 h; knockdown of Mpp53 decreased the survival rate of aphids under UV-B stress, suggesting that Mpp53 is involved in aphid responses to UV-B. Here, we first predicted 8 miRNAs targeting Mpp53, and then screened for miRNAs related to UV-B resistance in aphids; of these, 5 miRNAs (miR-305-5p, novel_50, novel_80, novel_166, and novel_61) were found to target Mpp53. Luciferase reporter assays demonstrated that novel_61 binds to the noncoding region of Mpp53 and downregulates its expression. Overexpression of novel_61 in aphids decreased Mpp53 expression and caused significant mortality under UV-B irradiation. Furthermore, the aphids exhibited lower reproductive capacity, lower body weight, and shorter body length and width. This is the first study to systematically screen and identify miRNA related to aphid responses to UV-B stress and deepens our understanding of the molecular mechanism of insect responses to environmental stress, which may eventually aid in developing better control strategies.

Antimicrobial peptides (AMPs) are critical components of innate immunity in diverse organisms, including plants, vertebrates, and insects. This study identified and characterized a novel Lepidoptera-specific AMP, named lepidoptin, from the invasive pest Spodoptera frugiperda (Lepidoptera: Noctuidae). Lepidoptin is a 116-amino acid protein containing a signal peptide and a novel β-sandwich domain that is distinct from previously reported AMPs. Temporal and spatial expression analyses revealed a significant upregulation of the lepidoptin gene in vivo and in cultured SF9 cells in response to pathogens. Molecular docking analysis identified a specific binding cavity. Enzyme-linked immunosorbent assay and binding assays confirmed that lepidoptin can bind to pathogen-associated molecular patterns, bacteria, and fungi. Recombinant lepidoptin exhibited potent antibacterial activity by inducing bacterial agglutination, inhibiting bacterial growth, increasing bacterial membrane permeability, and preventing biofilm formation. Lepidoptin also showed antifungal activity against the entomopathogenic fungus Beauveria bassiana by inhibiting spore germination, increasing fungal cell permeability, and increasing reactive oxygen species. Injection of recombinant lepidoptin into S. frugiperda larvae increased survival after B. bassiana infection, whereas knockdown of lepidoptin by RNA interference decreased larval survival. In addition, lepidoptin showed antimicrobial activity against the plant pathogen Fusarium graminearum by inhibiting spore germination and alleviating disease symptoms in wheat seedlings and cherry tomatoes. This study demonstrates the remarkable dual functionality of lepidoptin in enhancing S. frugiperda immunity and controlling plant pathogens, making it a promising candidate for biocontrol strategies in both pest management and plant disease prevention.

The use of synthetic pesticides carries a significant risk of pests developing resistance, leading to decreased pesticide effectiveness. ATP-binding cassette (ABC) transporters, especially the ABCC subfamily members, have been suggested to act as efflux pumps for various pesticides, thereby contributing to pesticide resistance. So far, the identification of potential pesticide substrates of insect ABC transporters is most often based on the quantification of transcript in arthropods. Here, we screened and identified the potential pesticide substrates of ABCC-9C from Tribolium castaneum based on an in vitro ATPase activity assay. Together with affinity evaluation-, cytotoxicity analysis-, and RNA interference-based bioactivity tests, we revealed that the insecticides, carbofuran, and buprofezin, are potential substrates of TcABCC-9C. Additionally, we identified an amphipathic translocation channel in the transmembrane domain of TcABCC-9C formed by 8 transmembrane helices. Molecular docking suggested that both carbofuran and buprofezin bind at the same site within the translocation channel via hydrophobic interactions. These findings indicate that TcABCC-9C might play a critical role in multi-pesticide resistance, providing a potential target for managing pesticide resistance and laying the groundwork for future pest control strategies. Given the conservations among ABCC subfamily members, the experimental model we developed in this study can be also applied to identify the potential substrates of other ABCC transporters, as well as to predict insecticide resistance mediated by ABCC transporters.

Lipids perform a diverse and unique array of functions in insects. Lipases are key enzymes in lipid metabolism, and their metabolic products are crucial for development and reproduction of insects. Here, a total of 110 lipase genes were identified in the genome of Spodoptera frugiperda. Cluster analysis indicated that neutral lipases constitute the majority of lipases. Tissue expression profile analysis displayed that most lipase genes were highly expressed in the larval gut of S. frugiperda. Some lipases exhibited a diet-specific expression pattern, which implied their roles in host adaptation. Key domain analysis proved that none of the neutral lipases highly expressed in the gut has an integrated lid domain, while most lipases highly expressed in the fat body contained both the integrated lid domain and β9 loop, indicating the activity loss of neutral lipases in guts. The assay of triacylglycerol (TAG) hydrolytic activity confirmed that the gut had the lowest activity when compared to that of fat body and epidermis. Interestingly, the opposite TAG hydrolytic activity trends across mating were observed between adult males and females, implying that lipase played different roles in the reproduction of both sexes. In conclusion, neutral lipases lost TAG hydrolytic activity in S. frugiperda guts, but retained the activity in fat body. Neutral lipases would play vital roles in many physiological processes in insects, especially in insect reproduction, which provides palpable targets for novel insecticide development to control insect population growth.

Fungal pathogens produce secretory ribonuclease (RNase) T2 proteins during infection, which contribute to fungal virulence via their enzyme functions in degradation of host cell RNA. However, the details of those proteins entering the host cells are unclear. Our previous study demonstrated that the two secretory RNase T2 members, BbRNT2 and BbTrv, produced by the insect fungal pathogen Beauveria bassiana, caused cytotoxic damage to insect cells and contributed to fungal virulence. Here, the Spodoptera frugiperda ovarian epithelial cells (sf9 cells) were used as models to investigate the interactions of the two fungus-produced RNase T2 proteins with the insect cells. Two transmembrane proteins, an ABC transporter (SfABCG) and an Innexin 7-like protein (Sfinx), were identified from the sf9 cells as interacting with BbRNT2 and BbTrv, respectively, through protein immunoprecipitation, yeast-two hybrid tests and protein pull-down assays. Although a slight decrease in the sf9 cell viability was examined by transfection of RNA interference of SfABCG or Sfinx, the transfected cells displayed a dramatically decreased sensitivity to BbRNT2 or BbTrv, suggesting the requirement of the two transmembrane proteins for BbRNT2 and BbTrv to enter the insect cells. These results reveal a mechanism of the cytotoxic molecules, T2 RNases, produced by the fungal pathogen, entering the insect cells via interaction with specific insect cell transmembrane proteins and causing cytotoxic damage.

Wild bees are ecologically vital but increasingly threatened by anthropogenic activities, leading to uncertain survival and health outcomes in urban environments. The gut microbiome contains features indicating host health and reflecting long-term evolutionary adaptation and acute reactions to real-time stressors. Moving beyond bacteria, we propose a comprehensive analysis integrating diet, bacteriome, virome, resistome, and their association to understand the survival status of urban lives better. We conducted a study on mason bees (Osmia excavata) across 10 urban agricultural sites in Suzhou, China, using shotgun gut metagenome sequencing for data derived from total gut DNA. Our findings revealed that most ingested pollen originated from Brassica crops and the unexpected garden tree Plantanus, indicating that floral resources at the 10 sites supported Osmia but with limited plant diversity. Varied city landscapes revealed site-specific flowers that all contributed to Osmia sustenance. The gut bacterial community, dominated by Gammaproteobacteria, showed remarkable structural stability across 8 sites but suggested perturbations at 2 sites. Antibiotic resistance gene profiles highly varied across 10 sites with prevalent unclassified drug classes, highlighting environmental threats to both bees and humans. The virome analysis identified honeybee pathogens, suggesting potential virus spillover. Many unknown bacteriophages were detected, some of which targeted the core gut bacteria, underscoring their role in maintaining gut homeostasis. These multifaceted metagenomic insights hold the potential to predict bee health and identify environmental threats, thereby guiding probiotic development and city management for effective bee conservation.