Insect Molecular Biology最新文献

The Hox gene Sex combs reduced (Scr) is recognized as a key factor in the development of the head and thorax in insects. However, its function in the growth, development and morphogenesis of Gryllus bimaculatus remains poorly understood. This study aimed to explore the function of the Scr gene in G. bimaculatus by using CRISPR/Cas9 technology to generate an Scr gene knock-out strain. Intercrossing the G₀ generation knock-out individuals with wild-type individuals yielded the G₁ generation to screen the mutant strain. It was found that the knock-out of the Scr gene had a severe impact on the growth and development of G. bimaculatus, resulting in high mortality and making it difficult to obtain Scr^-/- mutants. Therefore, heterozygous individuals (Scr^+/-) with 1 bp deleted were obtained for investigation. The results showed that the Scr deletion led to ectopic segment formation in the G₀ generation. In the G₂ generation, it was observed that stable Scr^-/- strains displayed abnormal embryonic development, characterized by enlarged, blackened and lethal eggs during embryogenesis. During the post-embryonic stage, Scr^-/- mutants exhibited abnormalities in body segmentation, particularly in the head-thorax region, resulting in a dorsal ridge structure. Furthermore, some Scr^+/- individuals exhibited a dorsal ridge during the nymphal stage. Notably, this characteristic did not persist into the adult stage. Our findings highlight the distinct but crucial roles of the Scr gene in both embryonic and post-embryonic growth and development of G. bimaculatus.

RNA-based bioinsecticides that comprise a dsRNA active ingredient and function by RNA interference (RNAi) are being commercialised as insecticidal traits in transgenic crops and as sprayable biopesticides. These RNAi insecticidal technologies are valuable alternatives to conventional chemical insecticides due to their efficacy, high degree of specificity and favourable human and environmental safety profiles. As with all pesticides, appropriate insect resistance management (IRM) programmes are required to mitigate the selection for resistance in target insect populations and extend product durability in the field. IRM programmes for RNAi products follow the same guidelines that currently exist for insecticidal traits or conventional insecticidal sprays. These guidelines reflect the distinct exposure scenarios for traits versus sprays, that is, continuous exposure when dsRNA is expressed in the crop compared to intermittent exposure when sprayed on foliage. As such, IRM plans for dsRNA traits depend on pyramiding (stacking) non-cross-resistant traits along with a refuge of non-transgenic plants. On the other hand, IRM plans for dsRNA sprays rely on the timing of the application so that only a single generation of the pest is exposed, followed by the use of an insecticide from a different IRAC mode of action group.

Spodoptera frugiperda (fall armyworm) poses a substantial risk to crops worldwide, resulting in considerable economic damage. The gut microbiota of insects plays crucial roles in digestion, nutrition, immunity, growth and, sometimes, the degradation of insecticides. The current study examines the effect of synthetic insecticides on the gut microbiome of third instar S. frugiperda larvae using both culture-dependent techniques and 16S rRNA gene sequencing for bacterial community profiling and diversity analysis. In untreated larvae, the sequencing approach revealed a diverse microbiome dominated by the phyla Firmicutes, Proteobacteria and Bacteroidota, with key genera including Bacteroides, Faecalibacterium and Pelomonas. In parallel, 323 bacterial strains were isolated and assigned to the orders Bacillales, Burkholderiales, Enterobacterales, Flavobacteriales, Lactobacillales, Micrococcales, Neisseriaies, Pseudomonadales, Sphingobacteriales and Xanthomonadales. The prevailing culturable species included Serratia marcescens, Klebsiella variicola and Enterobacter quasiroggenkampii. Treatment with sublethal concentrations of three insecticides (broflanilide, spinosad and indoxacarb) caused significant changes in gut microbiome diversity and composition. Treated larvae showed a shift towards increased Proteobacteria abundance and decreased Firmicutes. Specifically, Acinetobacter and Rhodococcus were dominant in treated samples. Functional predictions highlighted significant metabolic versatility involving nutrient processing, immune response, detoxification, xenobiotic metabolism, and stress response, suggesting microbial adaptation to insecticide exposure. Network correlation analysis highlighted disrupted microbial interactions and altered community structures under insecticide treatment. These findings enhance our understanding of how insecticides impact the gut microbiota in S. frugiperda and may inform future strategies for managing pest resistance through microbiome-based approaches.

Insects are known to synthesise and secrete hundreds of unique defensive chemicals, including caustic acids, pungent phenolics and citrusy terpenes. Despite efforts to characterise the defensive chemistry of ground beetles (Coleoptera: Carabidae), our knowledge of semiochemical evolution within the family and how these compounds are biosynthesised remains limited. Few studies have demonstrated the likely biosynthetic precursors of select compounds in certain taxa and only one has demonstrated which genes may be involved in the biosynthesis of formic acid. Here, we characterise the defensive chemistry and generate defensive gland transcriptomes for ground beetle species representing two defensive chemical classes: the formic acid producer Platynus angustatus and the methacrylic acid producer Pterostichus moestus. Through comparative transcriptome analyses, we demonstrate that co-option of distinct primary metabolic pathways may be involved in formic acid and methacrylic acid biosynthesis in the defensive glands of these taxa. These results expand our knowledge of ground beetle defensive chemistry and provide additional evidence that co-option of existing primary metabolic pathways plays a major role in the evolution of ground beetle chemical defence.

Autophagy is a cellular mechanism that enhances cell survival in response to various stressors, including nutrient deprivation; however, it also plays a pivotal role in the regulation of programmed cell death. This study examined the effects of autophagy-related genes Atg3, Atg5 and Atg12 on apoptosis and autophagy during the degeneration of the posterior silk gland in Bombyx mori, employing RNA interference techniques. Apoptosis-specific markers and autophagic processes were evaluated in both control and treatment groups. The knockdown of all three genes resulted in a significant reduction in autophagy, modifications in the apoptosis process, aberrant expression of p53 and impaired lysosomal function. It was determined that Atg3 is involved in the regulation of intracellular mitochondrial homeostasis. Following the silencing of Atg5, evidence was obtained indicating the gene's role in regulating lysosomal pH. Notably, the loss of Atg3 and Atg5 was associated with an increase in apoptotic markers, whereas the silencing of Atg12 inhibited apoptosis. Elevated levels of the p53 transcription factor following gene silencing suggested a potential interaction between these genes and p53. Our findings further underscore the importance of autophagy-mediated cell death, involving Atg3, Atg5 and Atg12, in the proper progression of degeneration in the posterior silk gland. A comprehensive understanding of the molecular mechanisms that mediate the interaction between apoptosis and autophagy is essential for elucidating their roles in both physiological and pathological contexts.

Sex determination pathways regulate male and female-specific development and differentiation and offer potential targets for genetic pest management methods. Insect sex determination pathways are comprised of primary signals, relay genes and terminal genes. Primary signals of coleopteran, dipteran, hymenopteran and lepidopteran species are highly diverse and regulate the sex-specific splicing of relay genes based on the primary signal dosage, amino acid composition or the interaction with paternally inherited genes. In coleopterans, hymenopterans and some dipterans, relay genes are Transformer orthologs from the serine-arginine protein family that regulate sex-specific splicing of the terminal genes. Alternative genes regulate the splicing of the terminal genes in dipterans that lack Transformer orthologs and lepidopterans. Doublesex and Fruitless orthologs are the terminal genes. Doublesex and Fruitless orthologs are highly conserved zinc-finger proteins that regulate the expression of downstream proteins influencing physical traits and courtship behaviours in a sex-specific manner. Genetic pest management methods can use different mechanisms to exploit or disrupt female-specific regions of different sex determination genes. Female-specific regions of sex determination genes can be exploited to produce a lethal gene only in females or disrupted to impede female development or fertility. Reducing the number of fertile females in pest populations creates a male-biased sex ratio and eventually leads to the local elimination of the pest population. Knowledge on the genetic basis of sex determination is important to enable these sex determination pathways to be exploited for genetic pest management.

Starvation can induce autophagy and apoptosis in intestinal cells. To elucidate the underlying mechanisms, we investigated autophagy and apoptosis in the midgut of the model insect, silkworm (Bombyx mori), focusing on calcium homeostasis. The results indicated that the body weight of silkworms decreased, along with damage to the morphology of their digestive tracts and midguts after starvation treatment. Additionally, mitochondrial swelling, autophagy and apoptosis were observable. Further investigation revealed that starvation upregulated the transcription of Ca²⁺ release channel-associated genes (e.g., BmIP3R, BmRyR) but suppressed the expression of Ca²⁺ efflux genes (BmPMCA), resulting in Ca²⁺ overload in midgut cells and subsequent upregulation of BmCalpain transcription. In addition, starvation increased the transcription of key autophagy genes (BmATG5, BmATG7, BmATG8) and the expression of the LC3-II protein. Upon prolonged starvation, the NtATG5 protein levels increased, a process that facilitated the transition from autophagy to apoptosis. These results indicate that Ca²⁺ overload activates the calpain-mediated apoptosis pathway and promotes apoptosis of midgut cells. The present study reveals the significant role that Ca²⁺ plays in the occurrence and transformation of autophagy and apoptosis induced by starvation treatment, thus providing a new research strategy for investigating the damage caused by starvation in biological organisms.

Insect NF-κB-like factor, Relish, is activated by viral infection and induces the production of antiviral proteins. In this study, we performed a transcriptomic analysis of BmE cells expressing the active form of BmRelish (BmRelish_act) and identified BmVago-like as the most strongly-induced secreted-protein. Expression of BmVago-like was specifically triggered by Bombyx mori Nucleo Polyhedro Virus (BmNPV) infection and regulated by BmSTING-BmRelish pathway. Incubating the fresh culture of cells with supernatant medium of BmVago-like expressing cells or recombinant BmVago-like protein (rBmVago-like) significantly increased antiviral resistance. On the contrary, reducing the expression of Bmvago-like by RNA interference (RNAi) in BmE cells as well as in silkworm larvae impaired antiviral response. Furthermore, we constructed transgenic silkworm line over-expressing BmVago-like (BmVago-like^OV) and found they had markedly lower viral load and higher survival rate after BmNPV infection compared with the wild-type control. Co-immunoprecipitation assay showed Bmintegrin β1 interacts with BmVago-like and it was involved in BmVago-like mediated antiviral response. Finally, we found the expression level of signalling molecules in the JAK-STAT pathway increased in rBmVago-like-treated cells and BmVago-like^OV silkworm larvae but decreased in RNAi-treated cells. In summary, our research uncovered an inducible antiviral response in silkworm mediated by cytokine BmVago-like, which is the downstream effector of BmSTING-BmRelish pathway and functions as an antiviral cytokine.

Dermestes frischii Kugelann, 1792 is a storage pest worldwide, and is important for estimating the postmortem interval in forensic entomology. However, because of the lack of transcriptome and genome resources, population genetics and biological control studies on D. frischii have been hindered. Here, single-molecule real-time sequencing and next-generation sequencing were combined to generate the full-length transcriptome of the five developmental stages of D. frischii, namely egg, young larva, mature larva, pupa and adult. A total of 41,665 full-length non-chimeric sequences and 59,385 non-redundant transcripts were generated, of which 42,756 were annotated in public databases. Using the weighted gene co-expression network analysis, gene co-expression modules related to the five developmental stages were constructed and screened, and the genes in these modules were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. The expression patterns of the differentially expressed genes (DEGs) related to olfaction and insect hormone biosynthesis were also explored. Transcription of most odorant binding proteins was up-regulated in the adult stage, suggesting they are important for foraging in adults. Many genes encoding for the ecdysone-inducible protein were up-regulated in the pupal stage, may be mainly responsible for the tissue remodelling of metamorphosis. The results of the quantitative real-time polymerase chain reaction (qRT-PCR) were consistent with the RNA-seq results. This is the first full-length transcriptome sequencing of dermestids, and the data obtained here are vital for understanding the stage-specific development and olfactory system of D. frischii, providing valuable resources for storage pest and forensic research.

Adipokinetic hormone (AKH), a crucial neuropeptide, participates in the important physiological processes by specially binding to its receptor to activate the AKH signalling pathway. AKH regulates energy metabolism. However, it remains unknown whether AKH affects larval development and adult reproduction by influencing energy metabolism. In the present study, the AKH was identified from Panonychus citri and contained the conserved functional domain 'Q-[LIV]-[NT]-F-[ST]-X (2)-W' that characterises the AKH family. The relative expression levels of PcAKH revealed different patterns of AKH expression at different developmental stages of P. citri. Feeding of double-standard RNA against PcAKH induced decreased fecundity and reduced survival, which was accompanied by the down-regulation of vitellogenin gene expression. In addition, after silencing the PcAKH, lipid metabolism and carbohydrate homeostasis were disrupted, manifested by increased body width and weight, and fasting phenomenon. Further investigation found that compared with the control, physiological changes in trehalose and triglyceride contents were accompanied by variations in the mRNA expression levels of genes related to lipid metabolism and carbohydrate metabolism. The disorder of lipid and carbohydrate metabolism may affect adult female reproduction, which may lead to insufficient vitellogenin deposition. Moreover, the silencing of PcAKH seriously affected the growth and development of larvae, which was manifested as delayed development period and difficulty in moulting. Conclusively, all these results in current study demonstrated that double-stranded RNA silencing system targeting PcAKH effectively inhibited larval development and female fecundity by disturbing lipid and carbohydrate metabolism, and PcAKH is a specific RNAi target for control of P. citri in the design and development of biopesticide in sustainable agriculture.