Insect Science最新文献

Serpins (serine protease inhibitors) constitute a superfamily of proteins with functional diversity and unusual conformational flexibility. In insects, serpins act as multiple inhibitors, by forming inactive acyl-enzyme complexes, in regulating Spätzles activation, phenoloxidases (POs) activity, and other cytokines. In this study, we present the cloning and characterization of Octodonta nipae serpin2 (OnSPN2), a 415 residues protein homologous to Tenebrio molitor 42Dd-like. Notably, OnSPN2 features an arginine residue (R364) at the P1 position, and additional arginine residues (R362, R367) at the P3 and P3' positions, respectively which is crucial for protease inhibition. Immunohistochemistry (IHC) and Western blot analyses revealed that OnSPN2 is primarily synthesized in plasmatocytes and then released into the plasma to exert its function. RNA interference results indicated that OnSPN2 knockdown may depress serine protease in melanization and remarkably increase the transcript level of Attacin in hemolymph, but its messenger RNA levels were not changed upon immune induction. Reciprocal co-immunoprecipitation assay results confirmed that OnSPN2 binds to OnPPAF1 and OnSP8, indicating its role as a negative regulator in the PO and AMP pathway. Intriguingly, several cathepsin-L isoforms were identified in the OnSPN2 immunoprecipitated samples. The cathepsin-L inhibition assays and protein-protein docking results, identified cathepsin-L as a potential target of OnSPN2. These results indicate that OnSPN2 is produced as an intracellular resident and additionally is associated with the PO and AMP pathway. OnSPN2 represents a multiple defense tool that may provide multiple antiproteolytic functions.

Cuticular proteins are essential for cuticle formation, molting, and survival in insects. However, functional analysis of cuticular proteins in the melon aphid has been limited. In this study, we identified an endocuticle structural glycoprotein (ESG) AgSgAbd-2-like in the melon aphid Aphis gossypii, which is a member of the RR-1 subfamily of the CPR (cuticular protein containing the conserved Rebers-Riddiford motif) chitin-binding proteins. When double-stranded RNA is delivered epidermally, AgSgAbd-2-like is knocked down, resulting in molting defects and mortality. The expression of AgSgAbd-2-like is comparatively low prior to molting and increases following molting. Ecdysone signaling consistently suppresses AgSgAbd-2-like. Histologically, the endocuticle and whole cuticle are thinner in AgSgAbd-2-like RNA interference (RNAi) aphids, which is a leading cause of molting defects and mortality. Furthermore, knockdown of any other homolog of ESGs, including AgSgAbd-4, AgSgAbd-4-like, AgSgAbd-8-like, and AgSgAbd-9-like, results in molting defects and death, like that by AgSgAbd-2-like RNAi. These results indicate that the melon aphid ESGs are conserved in cuticle formation and could be potential targets for RNAi-based pest management.

Succession is one of the most extensively studied ecological phenomena, yet debates persist about the importance of dispersal and external factors in driving this process. We aimed to quantify the influence of these factors by investigating how wing-related traits evolve across succession of blowfly (Diptera: Calliphoridae) communities in South Brazil. Rat carrion was placed in both forest and grassland habitats, and the associated blowfly communities were documented throughout the decomposition process. Using morphometric analysis, we measured wing and thorax traits and assessed trait changes over succession through mixed models. Our findings revealed that carrion succession follows distinct trajectories in forest and grassland environments. Specifically, we observed that Calliphora lopesi predominantly visited carcasses during the final phase of decomposition, resulting in significant differences in species composition and wing size between habitats. In forests, wing size increased toward the later stages of succession, whereas an opposite trend was observed in grasslands. Notably, these trait patterns were only evident at the species level, indicating that intraspecific trait variation is irrelevant. Stronger dispersers tend to arrive during the later stages of succession, suggesting that dispersal has a negligible role in shaping successional dynamics. Instead, environmental differences between habitats drive trait patterns throughout succession. Our results suggest that community composition in ephemeral resources is governed by deterministic processes and that successional stages can be predicted based on blowfly wing traits. Specifically, the presence of the large-winged C. lopesi indicates late decay, while the small-winged Chrysomia albiceps and Lucilia eximia are indicative of early decay.

Wings are important organs of insects involved in flight, mating, and other behaviors, and are therefore prime targets for pest control. The formation of insect wings is a complex process that is regulated by multiple pathways. The Hedgehog (Hh) pathway regulates the distribution of wing veins, while the Hippo pathway modulates wing size. Any interventions that can manipulate these pathways have the potential to disrupt wing development and could be used for pest control. In this study, we find that overexpression of miR-7 in Drosophila results in smaller wings with disordered veins. Mechanistically, miR-7 directly targets both ci and yki via different mature miRNAs (miR-7-5p and miR-7-3p), thereby disrupting the Hh and Hippo pathways. Importantly, this regulatory mechanism is also observed in another insect species, Helicoverpa armigera. Finally, by utilizing a nanocarrier delivery system, we show that introducing miR-7 via star polycation (SPc) leads to wing defects in H. armigera. In conclusion, these findings uncover that miR-7 inhibits wing formation by targeting both the Hippo and Hh pathways, indicating its potential for use in pest control strategies.

Many animals display physiological and behavioral activities limited to specific times of the day. Certain insects exhibit clear daily rhythms in their mating activities that are regulated by an internal biological clock. However, the specific genetic mechanisms underlying this regulation remain largely unexplored. Mating in the fruit fly Bactrocera dorsalis exhibits a daily rhythm and is dependent on sex pheromones produced in the male rectum. We used transcriptome sequencing and clustered regularly interspaced short palindromic repeats (CRISPR) / CRISPR-associated nuclease 9 techniques to understand whether the daily rhythmicity of mating in B. dorsalis and sex pheromone production in the rectum are regulated by clock genes. The results showed that the production of sex pheromones by B. dorsalis males is rhythmic (low during the day and high at night) and is influenced by clock genes. Knockout of the clock genes cryptochrome 1 (cry1) and timeless (tim) reduced the production of sex pheromones and significantly impaired mating ability in males. In addition, quantitative polymerase chain reaction results from 5 different tissues showed cry1 was highly expressed in the head, whereas tim was highly expressed in both the head and rectum (a key site for male sex pheromone production). Transcriptome analysis confirmed that cry1 (head) and tim (head and rectum) exhibit rhythmic expressions consistent with sex pheromone rhythmicity. These results suggest that cry1 may be related to a central clock neuron (like the suprachiasmatic nucleus), whereas the rhythmic expression of tim in the rectum indicates the potential presence of peripheral oscillators. Our study reveals new targets and ideas for improved control of the fruit fly.

Reproductive diapause is an insect survival strategy in which reproduction temporarily halts in response to adverse environmental changes. This process is characterized by arrested ovarian development and lipid accumulation in females. A reduction in juvenile hormone (JH) biosynthesis is known to initiate reproductive diapause, but its regulatory mechanism remains unclear. Seven up (Svp), a transcription factor from the nuclear receptor family, plays a crucial role in various developmental processes in insects. In this study, using the cabbage beetle Colaphellus bowringi as a model, we observed higher expression of Svp in the heads of female adults under reproductive photoperiodic conditions (short-day [SD]) compared to diapause conditions (long-day [LD]). RNA interference-mediated knockdown of Svp in SD females induced typical diapause phenotypes, including ovarian arrest and lipid accumulation. The application of methoprene (ME), a JH receptor agonist, reversed these diapause phenotypes and restored reproduction, indicating that Svp's regulation of reproductive diapause is dependent on JH signaling. Additionally, Svp knockdown led to the downregulation of JH pathway genes and a reduction in JH titers. Further evidence suggested that Svp regulates the expression of JHAMT1, a critical gene in JH biosynthesis, which determines diapause entry in C. bowringi. These findings suggest that diapause-inducing photoperiods suppress Svp expression, blocking JH production and triggering diapause. This work reveals a critical transcription factor that regulates reproductive diapause initiation through modulating JH production, providing a potential target for controlling pests capable of entering reproductive diapause.

Respiration is a vital process essential for organism survival, with most terrestrial insects relying on a sophisticated tubular tracheal network. In the current study, a gene with repetitive sequence was identified within the silkworm genome. Designated as BmMuc91C, it contains a dozen repeated motifs "PSSSYGAPX" and "GGYSSGGX" in its sequence. BmMuc91C exhibits specific expression in the tracheal system of silkworm larvae, with significantly higher expression levels during the molting stage. Overexpression of BmMuc91C in individual silkworms resulted in a marked increase in tracheal diameter, particularly during the molting stage. Immunofluorescence staining using a BmMuc91C antibody revealed a noticeable thickening of the apical extracellular matrix in the trachea. Tensile testing confirmed a considerable enhancement in tracheal elasticity. Additionally, a BmMuc91C mutation strain of silkworms was generated using the clustered regularly interspaced short palindromic repeats (CRISPR) / CRISPR-associated nuclease 9 system. Although no significant differences were observed in the growth, development, and molting of BmMuc91C mutant silkworms, mechanical tests demonstrated a decrease in tracheal elasticity. Transcriptomic techniques revealed that a significant number of cuticular and chitin-binding proteins were among the differentially expressed genes between mutant and wild-type silkworms. Furthermore, the recombined BmMuc91C protein was successfully expressed using the Escherichia coli system. Cross-linking experiments with horseradish peroxidase demonstrated the formation of macromolecular complexes of BmMuc91C, which exhibited spontaneous luminescent properties under ultraviolet light. This research sheds light on the role of elastic proteins in insect tracheae and provides valuable insights for the development of elastic biomaterials.

Polyphagous insect species develop using multiple host plants. Often considered beneficial, polyphagy can also be costly as host nutritional quality may vary. Drosophila suzukii (Matsumura) is an invasive species that can develop on numerous fruit species over the annual cycle. Here, we assessed the contribution of winter-available fruit to the development of seasonal populations of D. suzukii, under fluctuating late winter/early spring temperature regimes. We infested an artificial diet and three suitable fruit species available in winter/early spring (Aucuba japonica, Elaeagnus ×submacrophylla, Viscum album) with D. suzukii larvae under three temperature regimes: constant 20 °C, fluctuating controlled regime of 8-15 °C (12 h of light at 8 °C and 12 h of dark at 15 °C), and uncontrolled outdoor regime during spring. As expected, fly performance was impaired by early spring-like environmental conditions, whatever the development diet, and the winter fruit were suboptimal diets compared to the artificial diet, whatever the thermal regime. However, under cold fluctuating temperature regimes, the ranking of fruit supporting the best performance changed, highlighting the occurrence of physiological trade-offs. Winter-acclimated females preferentially oviposited in A. japonica and/or E. ×submacrophylla, whatever the thermal regime, which does not support the preference-performance hypothesis. This finding is also discussed in the context of D. suzukii management strategies.

Jewel beetles pose significant threats to forestry, and effective traps are needed to monitor and manage them. Green traps often catch more beetles, but purple traps catch a greater proportion of females. Understanding the function and mechanism of this behavior can provide a rationale for trap optimization. Jewel beetles possess UV-, blue-, green-, and red-sensitive photoreceptors, and perceive color differently from humans. Jewel beetle photoreceptor signals were calculated for tree leaf and tree bark stimuli, representing feeding and oviposition sites of adult jewel beetles respectively. Artificial neural networks (ANNs) were trained to discriminate those stimuli using beetle photoreceptor signals, providing in silico models of the neural processing that might have evolved to drive behavior. ANNs using blue-, green-, and red-sensitive photoreceptor inputs could classify these stimuli with very high accuracy (>99%). ANNs processed photoreceptor signals in an opponent fashion: increasing green-sensitive photoreceptor signals promoted leaf classifications, while increasing blue- and red-sensitive photoreceptor signals promoted bark classifications. Trained ANNs were fed photoreceptor signals calculated for traps, wherein they always classified green traps as leaves, but often classified purple traps as bark, indicating that these traps share salient features with different classes of tree stimuli from a beetle's eye view. A metric representing the photoreceptor opponent mechanism implicated by ANNs then explained catches of emerald ash borer, Agrilus planipennis, at differently colored traps from a previous field study. This analysis provides a hypothesized behavioral mechanism that can now guide the rational selection and improvement of jewel beetle traps.