Archives of Insect Biochemistry and Physiology最新文献

Insects have important symbiotic relationships with their intestinal microbiota. The intestinal microbiota is involved in or influences various processes in insects such as development, metabolism, immunity, and reproduction. Currently, research on the intestinal microbiota of parasitic insects is still in its early stages. The tachinid parasitoid Exorista sorbillans is a dipteran parasitic insect, with the silkworm (Bombyx mori) being its main host. Silkworms parasitized by E. sorbillans can suffer from severe silkworm maggot disease, which also poses a serious threat to sericulture. In this study, the intestinal microbiota of larval E. sorbillans at three instar stages was analyzed using 16S rRNA amplicon sequencing to explore the community composition of the intestinal microbiota. Additionally, using conventional culture methods, six cultivable strains were isolated and identified from the larval E. sorbillans on an antibiotic-free LB medium, and four cultivable strains were isolated and identified from the hemolymph of parasitized silkworms. This study investigated the E. sorbillans from the perspective of intestinal microbiota, elucidating the composition and structural characteristics of the intestinal microbiota of the tachinid parasitoid, and preliminarily discussing the functional roles of several major microorganisms, which helps to further clarify the potential mechanisms of interaction between the parasitoid and the silkworm.

Prostaglandin E₂ (PGE₂) is an eicosanoid that mediates a range of physiological actions in vertebrates and invertebrates, including reproduction and immunity. The PGE₂ receptor was identified and functionally assessed in two lepidopteran insects, Manduca sexta and Spodoptera exigua. However, its binding affinity to the receptor has not been reported. The PGE₂ receptor is a G-protein coupled receptor (GPCR) although its corresponding G-protein is not identified. PGE₂ binding assays were performed with membrane preparations from hemocytes of M. sexta larvae. We recorded an optimal binding in 4 h reactions conducted at pH 7.5 with 12 nM tritium-labeled PGE₂. We found that hemocytes express a single population of PGE₂ binding sites with a high affinity (Kd = 35 pmol/mg protein), which are specific and saturable. The outcomes of experiments on the influence of purine nucleotides suggested these are functional GPCRs. A bioinformatics analysis led to a proposed trimeric G-protein in the S. exigua transcriptome, in which the Gα subunit is classified into five different types: Gα(o), Gα(q), Gα(s), Gα(12), and Gα(f). After confirming expressions of these five types in S. exigua, individual RNA interference (RNAi) treatments were applied to the larvae using gene-specific double-stranded RNAs. RNAi treatments specific to Gα(s) or Gα(12) gene expression significantly suppressed the cellular immune responses although the RNAi treatments specific to other three Gα components did not. While PGE₂ treatments led to elevated hemocyte cAMP or Ca²⁺ levels, the RNAi treatments specific to Gα(s) or Gα(12) genes led to significantly reduced second messenger levels under PGE_2, although the RNAi treatments specific to the other three Gα components did not. These results showed that the PGE₂ receptor has high PGE₂ affinity in the nanomolar range and binds G-proteins containing a Gα(s) or Gα(12) trimeric component in S. exigua and M. sexta, and likely, all lepidopteran insects.

Furanocoumarins are a class of chemical compounds with phototoxic properties. For herbivores, efficient detoxification of such defense compounds is the prerequisite to feed successfully on furanocoumarin-containing plants. The cotton bollworm Helicoverpa armigera is a very important polyphagous pest in agriculture, but how it copes with toxic furanocoumarins in some of its host plants is not well understood. Given that cytochrome P450s are well known for their capacity in xenobiotic metabolism, this study attempted to explore the potential roles of cytochrome P450s in furanocoumarin transformation in this pest. Our data showed that two linear structures (psoralen and xanthotoxin) could be metabolized by three recombinant CYP9A enzymes, but no detectable depletion was observed for the linear one with the 8-dimethylallyloxy substituent on the coumarin moiety (imperatorin) and the angular furanocoumarin (angelicin). Initial epoxidation of the double bond connecting C2' and C3' of the furano ring following by cleavage of the epoxidated furan ring, leading to the formation of more soluble, less reactive and nonphotosensitizing metabolites, was identified as a common mechanism of linear furanocoumarin metabolism using a quadrupole/time-of-flight (Q-TOF) mass spectrometry interfaced with a high performance liquid chromatography (HPLC) system. Our data demonstrated that multiple P450s were involved in the detoxification of linear furanocoumarins in the cotton bollworm. These findings contribute to a better understanding of the biochemical basis of adaptation to plant defense chemicals in this economically important pest.

Presently, the European Commission is moving forward with a plan to reduce the use of chemical pesticides and increase the percentage of organic farming by 2030; this plan demands increased availability of biopesticides for organic farming. Glycoalkaloids are a class of molecules found in certain plants, including tomatoes and potatoes, which contain sugar and an alkaloid group. These compounds are known to have insecticidal properties, as they can act as natural defenses against insect pests. This study examined how a biological substance, glycoalkaloids, affects mortality and the retrocerebral complex of two beetle species, Tenebrio molitor and Zophobas atratus. More specifically, we focused on two glycoalkaloids, α-solanine and α-tomatine. Although these two insects belong to the same taxonomic family, they were not equally susceptible to these substances. In the mortality test, we observed that the T. molitor beetle seemed more susceptible, whereas the changes in the retrocerebral complex were more significant in the case of Z. atratus. In general, the effects vary between substances and insects, and changes in the retrocerebral complex may impair the reproduction of insects.

Short-wavelength blue light is ubiquitous in daily life and has a lasting destructive influence. Its potential harm to biological health is significant. This study used Drosophila as a model organism to investigate the protective effects of kaempferol, a flavonoid, against the toxicity of blue light. It also examined its physiological effects on Drosophila under blue light irradiation. In this experiment, fruit flies were fed with three different concentrations of kaempferol solutions (0.1, 0.01, and 0.001 mol/L) dissolved in food. The survival rate and physiological indexes of Drosophila were investigated under blue light irradiation of 2500 lux. The results showed that 0.1 mol/L kaempferol increased the activity of male flies during the day and significantly extended the male survival time under blue light irradiation. However, the study found that kaempferol did not significantly prolong the survival time of Drosophila in the oxidative stress experiment, and no significant difference was observed in the feeding experiment. In summary, our research found that kaempferol, at the concentration of 0.1 mol/L, has a protective effect on Drosophila under blue light irradiation, potentially achieved through alterations in circadian rhythm.

Phosphine fumigation is essential for controlling storage pests like Tribolium castaneum, but its frequent application has resulted in resistance, primarily due to mutations in the Dihydrolipoamide dehydrogenase (DLD) gene associated with the rph2 allele. This study demonstrates that the Patiala population exhibits homozygous resistance variations across populations, contrasting with the susceptibility observed in laboratory cultures. Our assessment of mitochondrial DLD and Cytochrome c oxidase (COX) activities showed significantly elevated levels in the Patiala population, with increases of approximately sevenfold for DLD and 6.92-fold for COX, indicating mitochondrial adaptations for enhanced energy production. Kinetic analyses of DLD in the resistant Patiala population showed a higher V_max (0.005 mmol/min) and a significantly increased Km (16.66 mM), indicating variations in maximal enzyme activity and substrate affinity. Furthermore, resistant T. castaneum populations displayed substantial upregulation of DLD and COX gene expression, with DLD expression increasing by 10.53-fold and COX expression peaking at 102.57-fold in Patiala. Pearson correlation analysis indicated strong positive correlations (r > 0.8) between enzymatic activity and gene expression for both DLD and COX, suggesting a coordinated role in resistance mechanisms. The PCA biplot illustrated distribution patterns of enzymatic activity and gene expression among field-resistant populations, highlighting the association between increased resistance and elevated enzymatic activity and gene expression levels. Therefore, the upregulation of DLD and COX activities in resistant populations underscores their critical roles in counteracting phosphine, reflecting metabolic reprogramming for improved energy production under stress.

The English grain aphid, Sitobion avenae, is a significant agricultural pest affecting wheat, barley, and oats. In Chile, the most prevalent and persistent clone (superclone) of S. avenae harbors the facultative endosymbiont bacterium Regiella insecticola. To determine the role of this bacterium in the reproductive success of this superclone, the presence of R. insecticola was manipulated to assess its impact on (1) the reproductive performance of this clone on two host plant species (wheat and barley), (2) the production of winged morphs, (3) changes in the insects' proteomic profiles, and (4) the root/shoot ratio of plant. It was found that the reproductive performance of this S. avenae superclone varied across host plants, depending on the presence of the facultative bacterial endosymbiont. Aphids infected with R. insecticola showed higher reproductive success on wheat, while the opposite effect was observed on barley. Aphid biomass was greater when infected with R. insecticola, particularly on barley. Additionally, aphids harboring R. insecticola exhibited a higher proportion of winged individuals on both host plants. Protein regulation in aphids on wheat was lower compared to those on barley. A higher root/shoot biomass ratio was observed in wheat plants compared to barley when infested by R. insecticola-infected aphid. Thus, R. insecticola significantly influences the reproductive performance and proteomic profile of a S. avenae superclone, with these effects shaped by the host plant. This suggests that the interaction between the host plant and the facultative endosymbiont contributes to the ecological success of this superclone.

An entomopathogenic nematode, Oscheius tipulae, was isolated from a soil sample. The identification of this species was supported by morphological and molecular markers. The nematode isolate exhibited pathogenicity against different target insects including lepidopteran, coleopteran, and dipteran insects. The virulence of this nematode was similar to that of a well-known entomopathogenic nematode, Steinernema carpocapsae, against the same insect targets. A comparative metagenomics analysis of these two nematode species predicted the existence of a combined total of 272 bacterial species in their intestines, of which 51 bacterial species were shared between the two nematode species. In particular, the common gut bacteria included several entomopathogenic bacteria including Xenorhabdus nematophila, which is known as a symbiotic bacterium to S. carpocapsae. The nematode virulence of O. tipulae to insects was enhanced by an addition of dexamethasone but suppressed by an addition of arachidonic acid, suggesting that the immune defenses of the target insects against the nematode infection is mediated by eicosanoids, which would be manipulated by the symbiotic bacteria of the nematode. Unlike S. carpocapsae, O. tipulae showed high virulence against dipteran insects including fruit flies, onion flies, and mosquitoes. O. tipulae showed particularly high control efficacies against the onion maggot, Delia platura, infesting the Welsh onion in the rhizosphere in both pot and field assays.

Soluble guanylate cyclase (sGC) serves as a receptor of nitric oxide (NO) and is the core metalloenzyme in the NO signal transduction pathway. sGC plays a key role in the NO-cGMP signal transduction pathway and participates in various physiological processes, including cell differentiation, neuron transmission, and internal environment homeostasis. sGC consists of two subunits, α and β, each subunit containing multiple isoforms. In this study, we cloned and analyzed the sGC-α1 gene in the silkworm Bombyx mori (BmsGC-α1). The BmsGC-α1 gene was expressed highest at the pupal stages. The highest BmsGC-α1 mRNA expression was observed in the head of fifth instar larvae and in fat body during the wandering stage of B. mori. Furthermore, we observed that feeding fifth instar larvae with thyroid hormone and nitroglycerin induced the expression of the BmsGC-α1 gene. Injection of BmsGC-α1 siRNA into silkworms at the prepupal stage resulted in a significant decrease in BmsGC-α1 expression levels at 48 and 72 h postinjection. After silencing BmsGC-α1, both the egg-laying amount and hatching rate of silkworm eggs were significantly reduced compared to the control group. These results suggest that BmsGC-α1 plays an important role in regulating the reproductive system of silkworms. This finding enhances our understanding of the functional diversity of sGC in insects.