The glutathione biosynthesis is involved in metamorphosis, antioxidant function, and insecticide resistance in Tribolium castaneum.

BACKGROUND: Reduced glutathione (GSH) synthesis is vital for redox homeostasis, cell-cycle regulation and apoptosis, and immune function. The glutamate-cysteine ligase catalytic subunit (Gclc) is the first and rate-limiting enzyme in GSH synthesis, suggesting the potential use of Gclc as a pesticide target. However, the functional characterization of Gclc, especially its contribution in metamorphosis, antioxidant status and insecticide resistance, is unclear in Tribolium castaneum. RESULTS: In this study, we identified and cloned Gclc from T. castaneum (TcGclc) and found that its expression began to increase significantly from the late larvae (LL) stage (3.491 ± 0.490-fold). Furthermore, RNA interference-mediated knockdown of TcGclc resulted in three types of aberration (100% total aberration rate) caused by the downregulation of genes related to the 20-hydroxyecdysone (20E) pathway. This deficiency was partially rescued by exogenous 20E treatment (53.1% ± 3.2%), but not by antioxidant. Moreover, in the TcGclc knockdown group, GSH content was decreased to 62.3%, and total antioxidant capacity, glutathione peroxidase and total superoxide dismutase activities were reduced by 14.6%, 83.6%, and 82.3%, respectively. In addition, treatment with different insecticides upregulated expression of TcGclc significantly compared with a control group during the late larval stage (P < 0.01). CONCLUSION: Our results indicate that TcGclc has an extensive role in metamorphosis, antioxidant function and insecticide resistance in T. castaneum, thereby expanding our understanding of GSH functions and providing a scientific basis for pest control. © 2024 Society of Chemical Industry.

Identification of potent inhibitors targeting Tribolium castaneum GSTe2 via structure-based screening and molecular dynamics simulation.

Red flour beetle, Tribolium castaneum, has a major negative impact during storage of agricultural products and reveals the negative impacts on human health. Insect-specific epsilon glutathione S-transferase (GSTs) which requires reduced glutathione (GSH) as an essential substrate not only develop insecticide resistance but also play important role in insect metamorphosis. Inhibition of the insect metamorphosis and the development of insecticide resistance could play an important role in pest control, so T. castaneum GSTe2 (TcGSTe2) in our previous study could be an important target protein for this purpose. This study aimed to find a potential TcGSTe2 inhibitors through in silico mothods, including molecular modeling, molecular docking, ADMET assay, followed by molecular dynamics (MD) simulation, principal component analysis and MM/PBSA analysis. The results showed that ZINC000169293362 and ZINC000095566957 were selected as potential TcGSTe2 inhibitors with high-binding affinity and without any toxicity from 3618 of GSH-like compounds obtained from ZINC database. MD simulation results revealed that TcGSTe2-ZINC000169293362 had more stability than that of reference GSH. Moreover, TcGSTe2-ZINC000169293362 and TcGSTe2-ZINC000095566957 showed lower binding free energy (-27.53 ± 0.16 kcal/mol and -18.83 ± 0.15 kcal/mol, respectively) compared with TcGSTe2-GSH (-8.90 ± 0.30 kcal/mol). This study could provide new insight into reduction of insecticide resistance and be used to design new inhibitors of insect GSTs.Communicated by Ramaswamy H. Sarma.

Gene family expansion analysis and identification of the histone family in Spodoptera frugiperda.

Spodoptera frugiperda, a major invasive pest, causes severe damage to various economically important crops. Previous comparative genomics studies have revealed a close association between the invasiveness of S. frugiperda and its genome. In recent years, a vast amount of genome from lepidopteran species has become available, offering an opportunity for a more detailed and comprehensive understanding of the biological characteristics of S. frugiperda. In this study, we conducted a comprehensive comparative genomics analysis of S. frugiperda using genome from 46 lepidopteran species. We found the highest number of gene family expansion events in S. frugiperda, indicating that gene family expansion is a crucial mechanism in its adaptive evolution. The expanded gene families are enriched in various biological processes, including nutrient metabolism, development, stress response, reproduction, and immune processes, suggesting that the expansion of these gene families likely contributes to the strong environmental adaptability of S. frugiperda. Furthermore, we identified the expansion of histone gene families in S. frugiperda which resulted from chromosome segmental duplications after the divergence from closely related species. Expression analysis of histone genes indicated that certain members might exert an influence on the growth and reproduction processes of S. frugiperda. Overall, our study deepens our understanding of the biological characteristics of S. frugiperda, providing a theoretical basis for the comprehensive management and sustained control of S. frugiperda and other lepidopteran pests in the future.

Functional characterisation of Fe (II) and 2OG-dependent dioxygenase TcALKBH4 in the red flour beetle, Tribolium castaneum.

Alpha-ketoglutarate-dependent dioxygenase ALKB homologue 4 (ALKBH4) is a member of the Fe (II) and 2-oxoglutarate-dependent ALKB homologue family that plays important roles in epigenetic regulation by alkyl lesions removal in mammals. However, the roles of ALKBH4 in insects are not clear. Here, TcALKBH4 was cloned and functionally characterised in Tribolium castaneum. Temporal expression revealed that TcALKBH4 was highly expressed in early embryos and early pupae. Spatial expression showed that TcALKBH4 was highly expressed in the adult testis, and followed by the ovary. RNA interference targeting TcALKBH4 at different developmental stages in T. castaneum led to apparent phenotypes including the failure of development in larvae, the reduction of food intake and the deficiency of fertility in adult. However, further dot blot analyses showed that TcALKBH4 RNAi does not seem to influence 6 mA levels in vivo. qRT-PCR was used to further explore the underlying molecular mechanisms; the result showed that TcALKBH4 mediates the development of larvae possibly through 20E signalling pathway, and the fertility of female and male adult might be regulated by the expression of vitellogenesis and JH signalling pathway, respectively. Altogether, these findings will provide new insights into the potential function of ALKBH4 in insects.

Insight into the structural and electrochemical properties of the interface between a Na6SOI2 solid electrolyte and a metallic Na anode.

Solid electrolytes (SE) have attracted a great deal of interest as they can not only mitigate the safety issues related to currently used liquid organic electrolytes but also enable the introduction of a metallic Na anode with extreme energy density in sodium-ion batteries. For such application, SE should exhibit high interfacial stability against metallic Na as well as high ionic conductivity, and Na6SOI2 with a Na-rich double anti-perovskite structure was recently identified as a promising SE candidate. In this work, we performed first principles calculations to investigate the structural and electrochemical properties of the interface between Na6SOI2 and a metallic Na anode. Our calculations revealed that interfaces could be formed safely, keeping the ultra-fast ionic conductivity of the bulk phase near the interface. Through the electronic structure analysis of the interface models, we found the change of upward valence band bending at the surface to downward band bending at the interface, being accompanied by electronic charge transfer from a metallic Na anode to Na6SOI2 SE at the interface. This work provides valuable atomistic insight into the formation and properties of the interface between SE and alkali metal for enhancing battery performance.

A novel GSTe2 involved in metamorphosis by regulating 20E signal pathway in Tribolium castaneum.

Insect-specific epsilon glutathion S-transferases (GSTs) are a class of multifunctional GST superfamily, which play important roles in detoxification of xenobiotic substances. Most research on GSTs has focused on insecticide detoxification and resistance, with little research on other physiological functions. Here, we identified and cloned the novel GSTe2 from Tribolium castaneum (TcGSTe2). Recombinant TcGSTe2 protein was successfully overexpressed in Escherichia coli and purified with affinity purification, which had high ability to catalyze the conjugation of reduced glutathione with 1-chloro-2,4-dinitrobenzene (CDNB). The expression level of TcGSTe2 was significantly decreased after exposure with four insecticides, phoxim, λ-cyhalothrin, dichlorvos, and carbofuran, in larval stage. Interestingly, RNA interference knockdown of TcGSTe2 caused metamorphosis deficiency in larval and pupal stages by inhibiting the 20E signal pathway. Furthermore, exogenous 20E injection partially rescued this metamorphosis deficiency and also increased the expression levels of 20E downstream response genes. This study illustrated TcGSTe2 plays an important role at metamorphosis beside the insecticide detoxification and resistance in T. castaneum.

C-doping into h-BN at low annealing temperature by alkaline earth metal borate for photoredox activity.

BCN (boron carbon nitride) nanosheets are promising photocatalyst materials for solar fuel production by visible light-driven water splitting and CO2 reduction due to their tunable band gap and unique properties. C-doping into h-BN by thermal annealing makes possible the preparation of BCN nanosheets with photocatalytic activity under visible light irradiation, but it generally requires a very high temperature (>1250 °C) from the thermodynamic viewpoint. Here, we report a new method to prepare BCN nanosheets with visible light-photocatalytic activity at lower annealing temperature (1000 °C) than equilibrium by adding alkaline earth metal compounds. BCN nanosheets formed in borate melt show a clear layered structure, tunable bandgap and photocatalytic activity for water splitting and CO2 reduction under visible light illumination. This provides a direction for doping other elements into h-BN at low annealing temperature by alkaline earth metal borates.