Bioscience Methods 2024, Vol.15, No.6, 302-314 http://bioscipublisher.com/index.php/bm 309 immune pathways such as RNAi and Jak-STAT in earwigs and other insects underscores the importance of these mechanisms in providing a robust defense against a wide array of pathogens (Kingsolver and Hardy, 2012). Furthermore, the integration and communication between different immune pathways, as seen in other insects, suggest that earwigs may also possess a highly coordinated and adaptable immune system (Zhang et al., 2021). In summary, while earwigs share many immune mechanisms with other insects, they also exhibit unique features and adaptations that reflect their specific ecological contexts and evolutionary history. Understanding these nuances can provide deeper insights into the diversity and evolution of insect immune systems. 8 Case Study 8.1 Examination of immune responses in Forficula auricularia The European earwig, Forficula auricularia, serves as an intriguing model for studying immune responses due to its ecological role and interactions with pathogens. Recent advancements in transcriptomic analysis have provided a comprehensive understanding of the genetic basis underlying these immune responses. A de novo transcriptome assembly of F. auricularia has revealed significant insights into the species' molecular biology, including the identification of over 8,800 contigs with significant similarity to insect-specific proteins. This extensive dataset has facilitated the assignment of Gene Ontology terms and the establishment of quantitative PCR tests for expression stability, which are crucial for studying immune-related gene expression (Roulin et al., 2014). 8.2 Pathogen challenges and immune activation Earwigs, like other insects, encounter a variety of pathogens in their natural habitats, necessitating robust immune defense mechanisms. Studies on Drosophila melanogaster have shown that infection with diverse pathogens induces the expression of numerous genes, many of which are involved in immune responses. For instance, a meta-analysis identified 62 genes significantly induced by infection, including those encoding known immune response factors and novel genes not previously associated with immunity. This highlights the complexity and specificity of immune responses to different pathogens (Waring et al., 2021). In the context of earwigs, similar pathogen challenges likely trigger the activation of immune pathways, although specific studies on earwig immune activation are still emerging. 8.3 Molecular analysis of immune-related gene expression under pathogen stress The molecular mechanisms underlying immune responses in earwigs involve the regulation of gene expression in response to pathogen exposure. In apple orchards, earwigs exposed to different pest management strategies exhibit variations in the expression of resistance-associated genes. For example, earwigs from organic orchards showed higher expression levels of acetylcholinesterase 2, a gene associated with resistance. Additionally, genes involved in detoxification, such as cytochromes P450, esterases, and glutathione S-transferases, were over-expressed in earwigs subjected to various management strategies (Figure 3). These findings underscore the impact of environmental factors on the molecular resistance mechanisms in earwigs and their potential role in biocontrol (Fricaux et al., 2023). The study of Fricaux et al. (2023) shows the relative expression levels of detoxification genes (CYP, CCE, and GST) in earwigs collected from various orchard treatments (no treatment, organic, integrated pest management, and conventional). The results indicate variable gene expression across different orchard treatments, with higher expression levels in treated orchards compared to non-treated ones. Significant increases in the expression of certain detoxification genes, such as CYP6NW1 and CYP6NQ1, are observed in treated groups, particularly in organic and integrated pest management conditions, indicating a response to pesticide exposure or other environmental factors. Furthermore, the role of Toll-like receptors (TLRs) in immune signaling pathways is well-documented in other organisms. TLRs detect conserved microbial structures and initiate immune responses, which include the induction of gene expression and coordination of gene regulation and metabolism. These mechanisms are likely conserved in earwigs, contributing to their ability to mount effective immune responses against pathogens (Fitzgerald and Kagan, 2020).
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