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Insect lytic polysaccharide monooxygenases (LPMO15s) are newly discovered copper‐dependent enzymes that promote chitin degradation in insect through oxidative cleavage of glycosidic bonds. They are potential pesticide targets due to their critical role for chitin turnover in the integument, trachea, and peritrophic matrix of the midgut during insect molting. However, the knowledge about whether and how LPMO15s participate in chitin turnover in other tissues is still insufficient. Here, using the orthopteran pest Locusta migratoria as a model, a novel alternative splicing site of LmLPMO15‐1 was discovered and it produces 2 variants, LmLPMO15‐1a and LmLPMO15‐1b. The transcripts of LmLPMO15‐1a and LmLPMO15‐1b were specifically expressed in the trachea and foregut, respectively. RNA interference targeting LmLPMO15‐1 (a common fragment shared by both LmLPMO15‐1a and LmLPMO15‐1b), a specific region of LmLPMO15‐1a or LmLPMO15‐1b all significantly reduced survival rate of nymphs and induced lethal phenotypes with developmental stasis or molt failure. Ultrastructure analysis demonstrated that LmLPMO15‐1b was specifically involved in foregut old cuticle degradation, while LmLPMO15‐1a was exclusively responsible for the degradation of the tracheal old cuticle. This study revealed LmLPMO15‐1 achieved tissue‐specific functional differentiation through alternative splicing, and proved the significance of the spliced variants during insect growth and development. It provides new strategies for pest control targeting LPMO15‐1. [ABSTRACT FROM AUTHOR]

RNA N6-methyladenosine (m6A), as the most abundant modification of messenger RNA, can modulate insect behaviors, but its specific roles in aggregation behaviors remain unexplored. Here, we conducted a comprehensive molecular and physiological characterization of the individual components of the methyltransferase and demethylase in the migratory locust Locusta migratoria. Our results demonstrated that METTL3, METTL14 and ALKBH5 were dominantly expressed in the brain and exhibited remarkable responses to crowding or isolation. The individual knockdown of methyltransferases (i.e., METTL3 and METTL14) promoted locust movement and conspecific attraction, whereas ALKBH5 knockdown induced a behavioral shift toward the solitary phase. Furthermore, global transcriptome profiles revealed that m6A modification could regulate the orchestration of gene expression to fine tune the behavioral aggregation of locusts. In summary, our in vivo characterization of the m6A functions in migratory locusts clearly demonstrated the crucial roles of the m6A pathway in effectively modulating aggregation behaviors. [ABSTRACT FROM AUTHOR]

Low-density lipoprotein receptor-related protein 2 (LRP2) is a multifunctional endocytic receptor expressed in epithelial cells. In mammals, it acts as an endocytic receptor that mediates the cellular uptake of cholesterol-containing apolipoproteins to maintain lipid homeostasis. However, little is known about the role of LRP2 in lipid homeostasis in insects. In the present study, we investigated the function of LRP2 in the migratory locust Locusta migratoria (LmLRP2). The mRNA of LmLRP2 is widely distributed in various tissues, including integument, wing pads, foregut, midgut, hindgut, Malpighian tubules and fat body, and the amounts of LmLRP2 transcripts decreased gradually in the early stages and then increased in the late stages before ecdysis during the nymphal developmental stage. Fluorescence immunohistochemistry revealed that the LmLRP2 protein is mainly located in cellular membranes of the midgut and hindgut. Using RNAi to silence LmLRP2 caused molting defects in nymphs (more than 60%), and the neutral lipid was found to accumulate in the midgut and surface of the integument, but not in the fat body, of dsLmLRP2-treated nymphs. The results of a lipidomics analysis showed that the main components of lipids (diglyceride and triglyceride) were significantly increased in the midgut, but decreased in the fat body and hemolymph. Furthermore, the content of total triglyceride was significantly increased in the midgut, but markedly decreased in the fat body and hemolymph in dsLmLRP2-injected nymphs. Our results indicate that LmLRP2 is located in the cellular membranes of midgut cells, and is required for lipid export from the midgut to the hemolymph and fat body in locusts.

Wings are an important flight organ of insects. Wing development is a complex process controlled by a series of genes. The flightless wing pad transforms into a mature wing with the function of migratory flight during the nymph-to-adult metamorphosis. However, the mechanism of wing morphogenesis in locusts is still unclear. This study analyzed the microstructures of the locust wing pads at pre-eclosion and the wings after eclosion and performed the comparative transcriptome analysis. RNA-seq identified 25,334 unigenes and 3,430 differentially expressed genes (DEGs) (1,907 up-regulated and 1,523 down-regulated). The DEGs mainly included cuticle development (LmACPs), chitin metabolism (LmIdgf4), lipid metabolism-related genes, cell adhesion (Integrin), zinc finger transcription factors (LmSalm, LmZF593 and LmZF521), and others. Functional analysis based on RNA interference and hematoxylin and eosin (H&E) staining showed that the three genes encoded zinc finger transcription factors are essential for forming wing cuticle and maintaining morphology in Locusta migratoria. Finally, the study found that the LmSalm regulates the expression of LmACPs in the wing pads at pre-eclosion, and LmZF593 and LmZF521 regulate the expression of LmIntegrin/LmIdgf4/LmHMT420 in the wings after eclosion. This study revealed that the molecular regulatory axis controls wing morphology in nymphal and adult stages of locusts, offering a theoretical basis for the study of wing development mechanisms in hemimetabolous insects.

Locust (Locusta migratoria) has a single striated muscle myosin heavy chain (Mhc) gene, which contains 5 clusters of alternative exclusive exons and 1 differently included penultimate exon. The alternative exons of Mhc gene encode 4 distinct regions in the myosin motor domain, that is, the N‐terminal SH3‐like domain, one lip of the nucleotide‐binding pocket, the relay, and the converter. Here, we investigated the role of the alternative regions on the motor function of locust muscle myosin. Using Sf9‐baculovirus protein expression system, we expressed and purified 5 isoforms of the locust muscle myosin heavy meromyosin (HMM), including the major isoform in the thorax dorsal longitudinal flight muscle (FL1) and 4 isoforms expressed in the abdominal intersegmental muscle (AB1 to AB4). Among these 5 HMMs, FL1‐HMM displayed the highest level of actin‐activated adenosine triphosphatase (ATPase) activity (hereafter referred as ATPase activity). To identify the alternative region(s) responsible for the elevated ATPase activity of FL1‐HMM, we produced a number of chimeras of FL1‐HMM and AB4‐HMM. Substitution with the relay of AB4‐HMM (encoded by exon‐14c) substantially decreased the ATPase activity of FL1‐HMM, and conversely, the relay of FL1‐HMM (encoded by exon‐14a) enhanced the ATPase activity of AB4‐HMM. Mutagenesis showed that the exon‐14a‐encoded residues Gly474 and Asn509 are responsible for the elevated ATPase activity of FL1‐HMM. Those results indicate that the alternative relay encoded by exon‐14a/c play a key role in regulating the ATPase activity of FL1‐HMM and AB4‐HMM. [ABSTRACT FROM AUTHOR]

This study investigated the influence of different linearly polarized spectrum lights on locusts polartactic response characteristics linearly polarized vector sensitivity mode and polartactic response) by using linearly polarized spectrum vector light module and experimental device. The objective was to clarify the vector sensitivity characteristics and functional effect of linearly polarized light spectrum intensity on locusts polartactic response, determine the influence specificity of linearly polarized spectrum illumination properties on locusts polarization-related behavior. When spectrum and illumination were constant, locusts polartactic response, presenting the response feature of sine and cosine function change specificity, was related to spectrum attribute. The visual acuity effect stimulated by violet spectrum was the best, whereas the optical distance modulation effect induced by orange spectrum was the strongest. When illumination was enhanced, locusts vector sensitivity mode shifted to present the specific sensitivity prompted by light intensity at long distance and inhibited by light intensity at short distance. Moreover, the regulating function of violet spectrum was the strongest, and the regulatory mutation effect of orange spectrum was the least significant. Simultaneously, locusts polartactic sensitivity to 300° vector at 100 lx, whereas to 240° vector at 1000 lx of linearly polarized violet light was the strongest. Locusts polartactic aggregation and visual tendency sensitivity to 90° vector at 100 lx, whereas to 270° vector at 1000 lx of linearly polarized violet light was the strongest. The heterogeneous regulation function of different linearly polarized spectrum couplings with light intensity led to significant variations in locusts vector sensitivity mode. This was derived from the antagonistic and specific tuning characteristics of locusts polartactic vision, reflecting the integrated output effect of locusts vector dependence regulated by linearly polarized spectrum intensity attribute. The findings were significant for the construction of pest polarization induction light sources and the investigation of the sensitive physiology pathway of locusts polarization vision. [ABSTRACT FROM AUTHOR]

Simple Summary: Fushi Tarazu Factor-1 (FTZ-F1) is a member of the nuclear receptor superfamily, which plays an important role in the molting process of insects. In this study, two FTZ-F1 transcripts (LmFTZ-F1-X1 and LmFTZ-F1-X2) were identified in Locusta migratoria, which were highly expressed in the cuticle. Then, the function of LmFTZ-F1-X1 and LmFTZ-F1-X2 in the molting process was explored using RNA interference. Silencing LmFTZ-F1-X1 and LmFTZ-F1-X2 separately did not affect the normal development of the nymph, but the simultaneous RNAi of LmFTZ-F1-X1 and LmFTZ-F1-X2 blocked the nymphal molting by regulating the genes involved in cuticle formation, chitin synthesis, and other key molting processes. Fushi-tarazu factor 1 (FTZ-F1) is a class of transcription factors belonging to the nuclear receptor superfamily and an important molting regulator in insects; however, its detailed function in the molting process of Locusta migratoria is still unclear. This study identified two FTZ-F1 transcripts (LmFTZ-F1-X1 and LmFTZ-F1-X2) in L. migratoria. The classical domains of FTZ-F1 were present in their protein sequences and distinguished based on their variable N-terminal domains. Reverse-transcription quantitative polymerase chain reaction analysis revealed that LmFTZ-F1-X1 and LmFTZ-F1-X2 were highly expressed in the integument. RNA interference (RNAi) was used to explore the function of LmFTZ-F1s in the molting of the third-instar nymph. Separate LmFTZ-F1-X1 or LmFTZ-F1-X2 silencing did not affect the normal development of third-instar nymphs; however, the simultaneous RNAi of LmFTZ-F1-X1 and LmFTZ-F1-X2 caused the nymphs to be trapped in the third instar stage and finally die. Furthermore, the hematoxylin-eosin and chitin staining of the cuticle showed that the new cuticles were thickened after silencing the LmFTZ-F1s compared to the controls. RNA-seq analysis showed that genes encoding four cuticle proteins, two chitin synthesis enzymes, and cytochrome P450 303a1 were differentially expressed between dsGFP- and dsLmFTZ-F1s-injected groups. Taken together, LmFTZ-F1-X1 and LmFTZ-F1-X2 are involved in the ecdysis of locusts, possibly by regulating the expression of genes involved in cuticle formation, chitin synthesis, and other key molting processes. [ABSTRACT FROM AUTHOR]

ABSTRACT Locusta migratoria is an important phytophagous pest, and its gut microbial communities play an important role in cellulose degradation. In this study, the gut microbial and cellulose digestibility dynamics of Locusta migratoria were jointly analyzed using high-throughput sequencing and anthrone colorimetry. The results showed that the gut microbial diversity and cellulose digestibility across life stages were dynamically changing. The species richness of gut bacteria was significantly higher in eggs than in larvae and imago, the species richness and cellulose digestibility of gut bacteria were significantly higher in early larvae (first and second instars) than in late larvae (third to fifth instars), and the diversity of gut bacteria and cellulose digestibility were significantly higher in imago than in late larvae. There is a correlation between the dynamics of gut bacterial communities and cellulose digestibility. Enterobacter, Lactococcus, and Pseudomonas are the most abundant genera throughout all life stages. Six strains of highly efficient cellulolytic bacteria were screened, which were dominant gut bacteria. Carboxymethyl cellulase activity (CMCA) and filter paper activity (FPA) experiments revealed that Pseudomonas had the highest cellulase enzyme activity. This study provides a new way for the screening of cellulolytic bacteria and lays the foundation for developing insects with significant biomass into cellulose-degrading bioreactors.IMPORTANCECellulose is the most abundant and cheapest renewable resource in nature, but its degradation is difficult, so finding efficient cellulose degradation methods is an urgent challenge. Locusta migratoria is a large group of agricultural pests, and the large number of microorganisms that inhabit their intestinal tracts play an important role in cellulose degradation. We analyzed the dynamics of Locusta migratoria gut microbial communities and cellulose digestibility using a combination of high-throughput sequencing technology and anthrone colorimetry. The results revealed that the gut microbial diversity and cellulose digestibility were dynamically changed at different life stages. In addition, we explored the intestinal bacterial community of Locusta migratoria across life stages and its correlation with cellulose digestibility. The dominant bacterial genera at different life stages of Locusta migratoria were uncovered and their carboxymethyl cellulase activity (CMCA) and filter paper activity (FPA) were determined. This study provides a new avenue for screening cellulolytic bacteria and lays the foundation for developing insects with significant biomass into cellulose-degrading bioreactors.

Female adults of the migratory locust, Locusta migratoria manilensis (Meyen), can sense seasonal photoperiod changes, which induces embryonic diapause as a key strategy to overwinter. Serine protease inhibitor genes (SPNs) were thought to play key roles during diapause, while few SPNs were functionally characterized. LmSPN2 was one of those genes differentially expressed between diapause and non-diapause eggs; however, its biological function remained to be explored. So, we conducted RNAi knockdown of LmSPN2, resulting in a significant decrease of the egg diapause rate by 29.7%. Using yeast two-hybrid assays, co-immunoprecipitation, and pull-down methods, we found an interaction between LmSPN2 and LmSPN3, which was proved to be mediated by a glutamate (E331) binding site of LmSPN2. RNAi knockdown of LmSPN3 resulted in a significant increase in diapause rate by 14.6%, indicating an inverse function of LmSPN2 and LmSPN3 on diapause regulation. Double knockdown of two SPN genes resulted in a 26.4% reduction in diapause rate, indicating that LmSPN2 was the dominant regulatory signal. Moreover, we found four Toll pathway genes (easter, spätzle, pelle, and dorsal) upregulated significantly after the knockdown of LmSPN2 while downregulated after the knockdown of LmSPN3. Therefore, we speculate that two SPNs regulate diapause through the Toll pathway. Our results indicated that LmSPN2 positively regulates locust egg entry into diapause, while LmSPN3 is a negative regulator of embryonic commitment to diapause. Their interaction is mediated by the binding site of E331 and influences egg diapause through the Toll pathway. This mechanistic understanding of diapause regulation expands our understanding of insect developmental regulation and provides functional targets for developing locust management strategies.

RNA interference (RNAi) is a specific post-transcriptional gene-silencing phenomenon, which plays an important role in the regulation of gene expression and the protection from transposable elements in eukaryotic organisms. In Drosophila melanogaster, RNAi can be induced by microRNA (miRNA), endogenous small interfering RNA (siRNA), or exogenous siRNA. However, the biogenesis of miRNA and siRNA in these RNAi pathways is aided by the double-stranded RNA binding proteins (dsRBPs) Loquacious (Loqs)-PB, Loqs-PD or R2D2. In this study, we identified three alternative splicing variants of Loqs, namely Loqs-PA, -PB, and -PC in the orthopteran Locusta migratoria. We performed in vitro and in vivo experiments to study the roles of the three Loqs variants in the miRNA- and siRNA-mediated RNAi pathways. Our results show that Loqs-PB assists the binding of pre-miRNA to Dicer−1 to lead to the cleavage of pre-miRNA to yield matured miRNA in the miRNA-mediated RNAi pathway. In contrast, different Loqs proteins participate in different siRNA-mediated RNAi pathways. In exogenous siRNA-mediated RNAi pathway, binding of Loqs-PA or LmLoqs-PB to exogenous dsRNA facilitates the cleavage of dsRNA by Dicer−2, whereas in endogenous siRNA-mediated RNAi pathway, binding of Loqs-PB or Loqs-PC to endogenous dsRNA facilitates the cleavage of dsRNA by Dicer−2. Our findings provide new insights into the functional importance of different Loqs proteins derived from alternative splicing variants of Loqs in achieving high RNAi efficiency in different RNAi pathways in insects.

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(1) Background: Few studies have been carried out to appraise abamectin toxicity toward Locusta migratoria nymphs. (2) Methods: This study aimed to evaluate the cytotoxic effect of abamectin as an insecticide through examining the changes and damage caused by this drug, in both neurosecretory cells and midgut, using L. migratoria nymphs as a model of the cytotoxic effect. Histopathological change in the brain was examined in both normal and abamectin-treated fifth-instar nymphs. Neurosecretory cells (NSCs) were also examined where there were loosely disintegrated cells or vacuolated cytoplasm. (3) Results: The results showed distinct histological changes in the gastrointestinal tract of L. migratoria nymphs treated with abamectin, with significant cellular damage and disorganization, i.e., characteristic symptoms of cell necrosis, a destroyed epithelium, enlarged cells, and reduced nuclei. The observed biochemical changes included an elevation in all measured oxidative stress parameters compared to untreated controls. The malondialdehyde activities (MDAs) of the treated nymphs had a five- to six-fold increase, with a ten-fold increase in superoxide dismutase (SOD), nine-fold increase in glutathione-S-transferase (GST), and four-fold increase in nitric oxide (NO). (4) Conclusions: To further investigate the theoretical method of action, a molecular docking simulation was performed, examining the possibility that abamectin is an inhibitor of the fatty acid-binding protein Lm-FABP (2FLJ) and that it binds with two successive electrostatic hydrogen bonds. [ABSTRACT FROM AUTHOR]

Chemical signals from conspecifics are essential in insect group formation and maintenance. Migratory locusts use the aggregation pheromone 4-vinylanisole (4VA), specifically released by gregarious locusts, to attract and recruit conspecific individuals, leading to the formation of large-scale swarms. However, how 4VA contributes to the transition from solitary phase to gregarious phase remains unclear. We investigated the occurrence of locust behavioral phase changes in the presence and absence of 4VA perception. The findings indicated that solitary locusts require crowding for 48 and 72 h to adopt partial and analogous gregarious behavior. However, exposure to increased concentrations of 4VA enabled solitary locusts to display behavioral changes within 24 h of crowding. Crowded solitary locusts with RNAi knockdown of Or35, the specific olfactory receptor for 4VA, failed to exhibit gregarious behaviors. Conversely, the knockdown of Or35 in gregarious locusts resulted in the appearance of solitary behavior. Additionally, a multi-individual behavioral assay system was developed to evaluate the interactions among locust individuals, and four behavioral parameters representing the inclination and conduct of social interactions were positively correlated with the process of crowding. Our data indicated that exposure to 4VA accelerated the behavioral transition from solitary phase to gregarious phase by enhancing the propensity toward proximity and body contact among conspecific individuals. These results highlight the crucial roles of 4VA in the behavioral phase transition of locusts. Furthermore, this study offers valuable insights into the mechanisms of behavioral plasticity that promote the formation of locust swarms and suggests the potential for 4VA application in locust control. [ABSTRACT FROM AUTHOR]

[Objectives] To investigate the effects of tea saponin on detoxification and protective enzyme activity of Locusta migratoria. [Methods] The effects of different concentrations of tea saponin on the activity of glutathione-S-transferase (GST), carboxylatetasse (CarE), cytochrome P450 (CYP450), peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) were compared by injecting low, medium and high concentrations of tea saponin solution into the abdomen of adult L. migratoria and monitoring changes in detoxification and protective enzyme activity after 0, 12, 24 and 48 h. [Results] The activity of the three detoxification enzymes increased rapidly in the low concentration treatment group, but GST activity was subsequently inhibited, and CarE activity was highest in the medium concentration treatment group. The activity of CYP450 increased with tea saponin dosage. CarE and CYP450 activity increased significantly after 12 h, and the activity of GST increased significantly after 24 h. POD and CAT had the highest activity in the low-concentration treatment group, whereas SOD activity increased significantly in the medium concentration treatment group. Protective enzyme activity began to increase significantly after 24 h in the low concentration treatment group. POD and SOD activity continued to increase, but CAT activity decreased, after 48 h. [Conclusion] Tea saponin can affect the activity of both detoxifying and protective enzyme in Locusta migratoria. [ABSTRACT FROM AUTHOR]

Fushi-tarazu factor 1 (FTZ-F1) is a class of transcription factors belonging to the nuclear receptor superfamily and an important molting regulator in insects; however, its detailed function in the molting process of Locusta migratoria is still unclear. This study identified two FTZ-F1 transcripts (LmFTZ-F1-X1 and LmFTZ-F1-X2) in L. migratoria. The classical domains of FTZ-F1 were present in their protein sequences and distinguished based on their variable N-terminal domains. Reverse-transcription quantitative polymerase chain reaction analysis revealed that LmFTZ-F1-X1 and LmFTZ-F1-X2 were highly expressed in the integument. RNA interference (RNAi) was used to explore the function of LmFTZ-F1s in the molting of the third-instar nymph. Separate LmFTZ-F1-X1 or LmFTZ-F1-X2 silencing did not affect the normal development of third-instar nymphs; however, the simultaneous RNAi of LmFTZ-F1-X1 and LmFTZ-F1-X2 caused the nymphs to be trapped in the third instar stage and finally die. Furthermore, the hematoxylin-eosin and chitin staining of the cuticle showed that the new cuticles were thickened after silencing the LmFTZ-F1s compared to the controls. RNA-seq analysis showed that genes encoding four cuticle proteins, two chitin synthesis enzymes, and cytochrome P450 303a1 were differentially expressed between dsGFP- and dsLmFTZ-F1s-injected groups. Taken together, LmFTZ-F1-X1 and LmFTZ-F1-X2 are involved in the ecdysis of locusts, possibly by regulating the expression of genes involved in cuticle formation, chitin synthesis, and other key molting processes.

The TRP channel superfamily was widely found in multiple species. They were involved in many extrasensory perceptions and were important for adapting to the environment. The migratory locust was one of the worldwide agricultural pests due to huge damage. In this study, we identified 13 TRP superfamily genes in the locust genome. The number of LmTRP superfamily genes was consistent with most insects. The phylogenetic tree showed that LmTRP superfamily genes could be divided into seven subfamilies. The conserved motifs and domains analysis documented that LmTRP superfamily genes contained unique characteristics of the TRP superfamily. The expression profiles in different organs identified LmTRP superfamily genes in the head and antennae, which were involved in sensory function. The expression pattern of different life phases also demonstrated that LmTRP superfamily genes were mainly expressed in third-instar nymphs and male adults. Our findings could contribute to a better understanding of the TRP channel superfamily gene and provide potential targets for insect control. [ABSTRACT FROM AUTHOR]

Hypoxia challenges aerobic organisms in numerous environments, and hypoxic conditions may become more severe under future climate‐change scenarios. The impact of hypoxia on the development of terrestrial insect embryos is not well understood. Here, to address this gap, embryonic life‐history traits of migratory locust Locusta migratoria from low‐altitude and high‐altitude regions were compared under 2 oxygen levels: normoxia (i.e., 21 kPa oxygen partial pressure and mild hypoxia (i.e., 10 kPa oxygen partial pressure). Our results demonstrated that, whether reared under normoxia or mild hypoxia, L. migratoria from high‐altitude populations had longer developmental times, reduced weight, and lower mean relative growth rate as compared with those from low‐altitude populations. When transferred from normoxia to mild hypoxia, nearly all the tested life‐history traits presented significant negative changes in the low‐altitude populations, but not in the high‐altitude populations. The factor 'strain' alone explained 18.26%–54.59% of the total variation for traits, suggesting that the phenotypic differences between L. migratoria populations from the 2 altitudes could be driven by genetic variation. Significant genetic correlations were found between life‐history traits, and most of these showed differentiation between the 2 altitudinal gradients. G‐matrix comparisons showed significant structural differences between L. migratoria from the 2 regions, as well as several negative covariances (i.e., trade‐offs) between traits in the low‐altitude populations. Overall, our study provides clear evidence that evolutionary divergence of embryonic traits between L. migratoria populations from different altitudes has occurred. [ABSTRACT FROM AUTHOR]

Locust and grasshopper plagues pose a serious threat to crop production in many areas worldwide. However, there is a lack of effective, quick-acting methods to control such outbreaks. Methods exploiting the phototactic response of these insects are receiving increasing attention. The current study investigated the effect of linearly polarized and unpolarized light on locust phototactic and polarotactic responses, in particular the function of their dorsal rim area (DRA) and non-DRA visual fields. The results showed that the polarotactic function weight of DRA vision was stimulated by linearly polarized ultraviolet (UV) and violet light, the phototactic function weight was induced by blue, green, and orange light, and under linearly polarized light, the functional effect of DRA vision was strongest in response to linearly polarized violet light. Moreover, the locust visual response effect was related to spectral light attributes, with the linear polarization effect intensifying in response to the short-range vision sensitivity of non-DRA visual fields, whereas DRA vision regulated the short-range sensitivity of compound eye vision. When illumination increased, the synergistic enhancement effects of linearly polarized ultraviolet and violet light were significant, whereas the visual sensitivity was restricted significantly by linearly polarized blue, green, or orange light. Thus, non-DRA vision determined, while DRA vision enhanced, the phototactic response sensitivity, whereas, in linearly polarized UV or violet light, non-DRA vision determined, while DRA vision enhanced, the visual trend and polarotaxic aggregation sensitivity, with opposite effects in linearly polarized blue, green, or orange light. When illumination increased, there was a driving effect caused by linearly polarized violet light on non-DRA vision, whereas at short-wave lengths, the control effect induced by linearly polarized orange light was optimal; however, the photo-induced effect of linearly polarized violet light and the visual distance control effect of linearly polarized orange light were optimal. These results provide theoretical support for the photo-induced mechanism of the locust visual response effect and for the development of linearly polarized light sources for the environmentally friendly prevention and control of locust populations. [ABSTRACT FROM AUTHOR]

Rising population in the world causes reduction on present food resources. Investigators have been looking for sustainable, nutritional and alternative food-stocks. Insects have been consumed as snacks or as food supplement in many countries, but they are still not attractive food resource worldwide. In the current study, β-fructofuranosidase enzyme production by Galactomyces geotrichum TS61 (GenBank accession: MN749818) strain was investigated using an edible insect (Locusta migratoria) as substrate. β-fructofuranosidase is a valuable enzyme in food industry. Taguchi L16 design of experiment (DOE) was employed to achieve an effective statistical optimization process, including three factors (concentration of locust powder, concentration of sucrose and initial pH) with four levels. The optimized conditions were determined as 40 g/L locust powder, 30 g/L sucrose and 6.0 pH. The analysis of variance results showed that locust powder had more effect on the enzyme production than sucrose and pH. At the end of the optimization process, approximately 4-fold higher β-fructofuranosidase production (40.91 U/mL) was obtained when compared with unoptimized experimental run (9.91 U/mL). Consequently, powdered insects may serve as an effective supplement for valuable enzyme production in food industry. [ABSTRACT FROM AUTHOR]

In this study, we shed light for the first time on the usage of migratory locusts (Locusta migratoria) as an insect model to investigate the nanotoxicological influence of aluminum oxide (Al2O3) nanoparticles at low doses on testes, and evaluate the capacity of a whole-body extract of American cockroaches (Periplaneta americana) (PAE) to attenuate Al2O3 NPs-induced toxicity. Energy dispersive X-ray microanalyzer (EDX) analysis verified the bioaccumulation of Al in testicular tissues due to its liberation from Al2O3 NPs, implying their penetration into the blood–testis barrier. Remarkably, toxicity with Al engendered disorders of antioxidant and stress biomarkers associated with substantial DNA damage and cell apoptosis. Furthermore, histopathological and ultrastructural analyses manifested significant aberrations in the testicular tissues from the group exposed to Al2O3 NPs, indicating the overproduction of reactive oxygen species (ROS). Molecular docking analysis emphasized the antioxidant capacity of some compounds derived from PAE. Thus, pretreatment with PAE counteracted the detrimental effects of Al in the testes, revealing antioxidant properties and thwarting DNA impairment and cell apoptosis. Moreover, histological and ultrastructural examinations revealed no anomalies in the testes. Overall, these findings substantiate the potential applications of PAE in preventing the testicular impairment of L. migratoria and the conceivable utilization of locusts for nanotoxicology studies. [ABSTRACT FROM AUTHOR]

Paranosema (Antonospora) locustae (Canning) is a microsporidium with an extensive host range including >100 reported insect hosts from the order Orthoptera. The susceptibility of two species of locusts (Orthoptera: Acrididae) – the migratory locust, Locusta migratoria subsp. migratorioides (Fairmaire & Reiche), and the desert locust, Schistocerca gregaria Forsskål – to P. locustae was compared under laboratory conditions at a decreased temperature of 27 °C to enhance susceptibility to infection. Locusta migratoria was found highly vulnerable as infection percentages exceeded 70% at 104, 105, and 106 spores per insect, and mortality increased with increasing dosage. Conversely, P. locustae spores were not found in S. gregaria throughout the experiment. Only at 107 spores per insect, as many as 20% of S. gregaria were infected. This suggests that the desert locust is resistant to P. locustae infection, as opposed to the migratory locust. The laboratory models of these parasite–host systems may be useful for elucidating mechanisms of insect immunity to microsporidia. [ABSTRACT FROM AUTHOR]

Lower plant resistance to herbivores following domestication has been suggested as the main cause for higher feeding damage in crops than in wild progenitors. While herbivore compensatory feeding has also been proposed as a possible mechanism for raised damage in crops with low nutritional quality, predictions regarding the effects of plant domestication on nutritional quality for herbivores remain unclear. In particular, data on primary metabolites, even major macronutrients, measured in the organs consumed by herbivores, are scarce. In this study, we used a collection of 10 accessions of wild ancestors and 10 accessions of modern progenies of Triticum turgidum to examine whether feeding damage and selectivity by nymphs of Locusta migratoria primarily depended on five leaf traits related to structural resistance or nutrient profiles. Our results unexpectedly showed that locusts favored wild ancestors over domesticated accessions and that leaf toughness and nitrogen and soluble protein contents increased with the domestication process. Furthermore, the quantitative relationship between soluble protein and digestible carbohydrates was found to poorly meet the specific requirements of the herbivore, in all wheat accessions, both wild and modern. The increase in leaf structural resistance to herbivores in domesticated tetraploid wheat accessions suggested that resource allocation trade‐offs between growth and herbivory resistance may have been disrupted by domestication in the vegetative organs of this species. Since domestication did not result in a loss of nutritional quality in the leaves of the tetraploid wheat, our results rather provides evidence for a role of the content of plants in nonnutritive nitrogenous secondary compounds, possibly deterrent or toxic, at least for grasshopper herbivores. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) is a specific post-transcriptional gene-silencing phenomenon, which plays an important role in the regulation of gene expression and the protection from transposable elements in eukaryotic organisms. In Drosophila melanogaster, RNAi can be induced by microRNA (miRNA), endogenous small interfering RNA (siRNA), or exogenous siRNA. However, the biogenesis of miRNA and siRNA in these RNAi pathways is aided by the double-stranded RNA binding proteins (dsRBPs) Loquacious (Loqs)-PB, Loqs-PD or R2D2. In this study, we identified three alternative splicing variants of Loqs, namely Loqs-PA, -PB, and -PC in the orthopteran Locusta migratoria. We performed in vitro and in vivo experiments to study the roles of the three Loqs variants in the miRNA- and siRNA-mediated RNAi pathways. Our results show that Loqs-PB assists the binding of pre-miRNA to Dicer−1 to lead to the cleavage of pre-miRNA to yield matured miRNA in the miRNA-mediated RNAi pathway. In contrast, different Loqs proteins participate in different siRNA-mediated RNAi pathways. In exogenous siRNA-mediated RNAi pathway, binding of Loqs-PA or LmLoqs-PB to exogenous dsRNA facilitates the cleavage of dsRNA by Dicer−2, whereas in endogenous siRNA-mediated RNAi pathway, binding of Loqs-PB or Loqs-PC to endogenous dsRNA facilitates the cleavage of dsRNA by Dicer−2. Our findings provide new insights into the functional importance of different Loqs proteins derived from alternative splicing variants of Loqs in achieving high RNAi efficiency in different RNAi pathways in insects. [ABSTRACT FROM AUTHOR]

Insects comprise more than a million species and many authors have attempted to explain this success by evolutionary innovations. A much overlooked evolutionary novelty of insects is the serosa, an extraembryonic epithelium around the yolk and embryo. We have shown previously that this epithelium provides innate immune protection to eggs of the beetle Tribolium castaneum. It remained elusive, however, whether this immune competence evolved in the Tribolium lineage or is ancestral to all insects. Here, we expand our studies to two hemimetabolous insects, the bug Oncopeltus fasciatus and the swarming grasshopper Locusta migratoria. For Oncopeltus, RNA sequencing reveals an extensive response upon infection, including the massive upregulation of antimicrobial peptides (AMPs). We demonstrate antimicrobial activity of these peptides using in vitro bacterial growth assays and describe two novel AMP families called Serosins and Ovicins. For both insects, quantitative polymerase chain reaction shows immune competence of the eggs when the serosa is present, and in situ hybridizations demonstrate that immune gene expression is localized in the serosa. This first evidence from hemimetabolous insect eggs suggests that immune competence is an ancestral property of the serosa. The evolutionary origin of the serosa with its immune function might have facilitated the spectacular radiation of the insects. This article is part of the theme issue 'Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom'. [ABSTRACT FROM AUTHOR]

Clathrin heavy chain (Chc) is a constituent of clathrin‐coated vesicles and serves important functions in endocytosis and intracellular membrane trafficking but appears to have physiological roles also at the organismal level. Most of what we know about Chc functions originates from studies performed in fungal or vertebrate cells. However, the physiological functions of Chc in insects remain poorly understood. Here, we identified a Chc ortholog from a Locusta migratoria transcriptome database. RT‐qPCR revealed that LmChc was constitutively expressed in fifth‐instar nymphs. In this developmental stage, LmChc showed the highest expression in the ovary followed by hemolymph, testis, hindgut, midgut, and foregut. In isolated hemocytes, we detected the Chc protein in patches at the plasma membrane. To examine the role of LmChc in L. migratoria during development, RNA interference was performed by injecting dsRNA into the early fifth‐instar nymphs. Silencing of LmChc caused a lethal phenotype with molting defect from nymph to adult. In addition, silencing of LmChc resulted in abnormal development of the ovaries, the size of which was significantly smaller than that in controls. Taken together, our results suggest that LmChc is a vital gene in L. migratoria that plays an important role in growth, development, and reproduction. LmChc may be used as an efficient RNAi target gene for developing dsRNA‐based biological insecticides to manage insect pests. [ABSTRACT FROM AUTHOR]

Grasshoppers can swarm in the millions and destroy crops over wide areas, posing a major economic threat to agriculture. A wide range of insect-related viruses has recently been reported in the metagenomics of grasshoppers. Here, we identified and isolated a novel reovirus from grasshoppers, named Acrididae reovirus (ARV). The complete genome of ARV was composed of nine dsRNA segments. Phylogenetic analysis revealed that ARV formed a monophyletic lineage with unclassified insect-associated reoviruses and was sufficiently distinct from known genera of Reoviridae. ARV could replicate in its host Locusta migratoria and result in host death. Lower-dose ARV infection affected ovary development and resulted in a significant reduction in fecundity. The identification and characterization of a novel pathogenic reovirus could potentially promote the development of new biological control agents. [ABSTRACT FROM AUTHOR]

The ovipositor comprises the external genitalia of female insects, which plays an important role in the mating and ovipositing process of insects. However, it remains rudimentary of regional gene expression and physiological function in the ovipositor during structural development. Here, we analysed the basic structure and characteristics of the ovipositor in the migratory locust Locusta migratoria. RNA‐seq analysis revealed the specialization of chitin metabolism, lipids synthesis and transport, tanning and cuticular protein genes in the ovipositor. Among them, two cuticle protein genes, LmCP8 and LmACP79, were identified, which are specifically expressed in the ovipositor. Functional analysis based on RNA interference showed that deficiency of LmCP8 affected the structural development of the ovipositor resulting in the retention of a large number of remaining unproduced oocysts in the ovary of the locusts. Our results provide a fundamental resource to investigate the structural development and physiological function of the ovipositor in L. migratoria. [ABSTRACT FROM AUTHOR]

Immunity and reproduction are vital functions for the survival and population maintenance of female insects. However, owing to limited resources, these two functions cannot be fulfilled simultaneously, resulting in an energy tradeoff between them. Notably, the mechanisms underlying this immune-reproductive trade-off, in which energy competition likely plays a central role, remain poorly understood. Fatty acid synthase (FAS), a key gene involved in lipid synthesis and insect energy metabolism, was investigated in this study using Locusta migratoria as the research subject. Bacterial infection and RNA interference (RNAi) technology were used to examine changes in the immunity, fecundity, and energy metabolism patterns of locusts under different treatments. The findings of this study demonstrate that infection with Micrococcus luteus triggers an immune response in locusts, significantly upregulates the expression of defensin 3 (DEF3) and Attacin , and enhances pHenoloxidase (PO) activity. Upon FAS2 silencing, bacterial attack upregulated DEF3 and Attacin expression to a lesser extent, leading to increased lysozyme activity instead of PO. Furthermore, bacterial infection results in a decrease in glycogen and glucose content in the fat body, accompanied by a significant increase in triacylglycerol (TAG) content. However, after FAS2 knockdown, both the lipid and carbohydrate contents were significantly reduced in the fat body. Compared with bacterial infection alone, low FAS2 expression further exacerbated fecundity impairment in locusts. The expression levels of vitellogenin A (VgA) and vitellogenin B (VgB) were significantly low, with severe ovarian atrophy observed. Notably, the ovarian weight was only 21 % compared to that of the control group. Moreover, females exhibited minimal egg-laying behavior. In summary, our findings suggest that following FAS2 gene silencing, there is a greater inclination toward immune stimulation energy activation in locusts, whereas reproductive investment is reduced. The outcomes of this study will contribute to the further exploration of the molecular mechanisms underlying the trade-off between immune and reproductive energy in locusts. [Display omitted] • M. luteus infection reduces the fecundity of locusts. • LmFAS2 plays an important role in lipid synthesis and reproduction. • dsFAS2 + M. luteus infection seriously impaired the fecundity of locusts. [ABSTRACT FROM AUTHOR]

(1) Background: Few studies have been carried out to appraise abamectin toxicity toward Locusta migratoria nymphs. (2) Methods: This study aimed to evaluate the cytotoxic effect of abamectin as an insecticide through examining the changes and damage caused by this drug, in both neurosecretory cells and midgut, using L. migratoria nymphs as a model of the cytotoxic effect. Histopathological change in the brain was examined in both normal and abamectin-treated fifth-instar nymphs. Neurosecretory cells (NSCs) were also examined where there were loosely disintegrated cells or vacuolated cytoplasm. (3) Results: The results showed distinct histological changes in the gastrointestinal tract of L. migratoria nymphs treated with abamectin, with significant cellular damage and disorganization, i.e., characteristic symptoms of cell necrosis, a destroyed epithelium, enlarged cells, and reduced nuclei. The observed biochemical changes included an elevation in all measured oxidative stress parameters compared to untreated controls. The malondialdehyde activities (MDAs) of the treated nymphs had a five- to six-fold increase, with a ten-fold increase in superoxide dismutase (SOD), nine-fold increase in glutathione-S-transferase (GST), and four-fold increase in nitric oxide (NO). (4) Conclusions: To further investigate the theoretical method of action, a molecular docking simulation was performed, examining the possibility that abamectin is an inhibitor of the fatty acid-binding protein Lm-FABP (2FLJ) and that it binds with two successive electrostatic hydrogen bonds.

Yu Wang,1,2 Yuan Zhang,3 Hongfei Lou,3 Chengshuo Wang,3 Ming Ni,1 Dan Yu,4 Luo Zhang,3 Le Kang1,5 1College of Life Science, Institute of Life Sciences and Green Development, Hebei University, Baoding, Hebei Province, People’s Republic of China; 2College of Food Science and Technology, Hebei Agricultural University, Baoding, Hebei Province, People’s Republic of China; 3Department of Otolaryngology Head and Neck Surgery, Beijing TongRen Hospital, Capital Medical University, Beijing, People’s Republic of China; 4State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China; 5Guangzhou Laboratory, Guangzhou, 510320, People’s Republic of ChinaCorrespondence: Le KangCollege of Life Science, Hebei University, Baoding, Hebei Province, People’s Republic of China, Email lkang@ioz.ac.cn; Luo ZhangDepartment of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, No. 17, Hougou Hutong, Dongcheng Beijing Institute of Otolaryngology, District, Beijing, 100730, People’s Republic of China, Email dr.luozhang@139.comBackground: Occupational exposure to locusts induces a high prevalence of allergic sensitization. However, knowledge on occupational locust allergens remains unclear.Objective: This study aimed to identify the allergens from locusts causing occupational allergies.Methods: We conducted a survey of 57 persons exposed to locusts using questionnaires and immunological tests for occupational allergies in long-term locust laboratories. The major allergen was identified by immunoblotting and analysed by mass spectrometry. The allergenicity of the allergen was assessed by sIgE detection, immunoblotting and ELISA inhibition assays.Results: The survey indicated that the frequency of locust occupational allergies was 40.4% among subjects exposed to locust. The symptoms in most males were allergic rhinitis, while females showed higher prevalence of atopic dermatitis. Occupational exposure increased the allergy risk. The recombinant hexamerin-2 protein possesses high allergenicity in the allergic exposure group. Hexamerin-2 protein can inhibit IgE reactivity with locust protein extracts by approximately 60%. The potential for cross-reactivity with cockroaches was indicated by sequence alignment of hexamerin-2 protein and allergens of cockroaches.Conclusion: The hexamerin-2 protein of locusts as an important allergen was identified. Therefore, occupational exposure is an important risk factor for locust allergy.Keywords: locust, Locusta migratoria, occupational allergy, hexamerin-2 protein, contact dermatitis, asthma

BACKGROUND: The selective aphicide flonicamid is known to cause symptoms in aphids that are like those of chordotonal organ TRPV channel modulator insecticides such as pymetrozine, pyrifluquinazon and afidopyropen. Flonicamid is classified by the Insecticide Resistance Action Committee as a chordotonal organ modulator with an undefined target site. However, although it has been shown not to act on TRPV channels, flonicamid's action on chordotonal organs has not been documented in the literature. RESULTS: Flonicamid causes locusts to extend their hindlegs, indicating an action on the femoral chordotonal organ. In fruit flies, it abolishes negative gravitaxis behavior by disrupting transduction and mechanical amplification in antennal chordotonal neurons. Although flonicamid itself only weakly affects locust chordotonal organs, its major animal metabolite 4‐trifluoromethylnicotinamide (TFNA‐AM) potently stimulates both locust and fly chordotonal organs. Like pymetrozine, TFNA‐AM rapidly increases Ca2+ in antennal chordotonal neurons in wild‐type flies, but not iav1 mutants, yet the effect is nonadditive with the TRPV channel agonist. CONCLUSIONS: Flonicamid is a pro‐insecticide form of TFNA‐AM, a potent chordotonal organ modulator. The functional effects of TFNA‐AM on chordotonal organs of locusts and flies are indistinguishable from those of the TRPV agonists pymetrozine, pyrifluquinazon and afidopyropen. Because our previous results indicate that TFNA‐AM does not act directly on TRPV channels, we conclude that it acts upstream in a pathway that leads to TRPV channel activation. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Lytic polysaccharide monooxygenases (LPMOs) are important enzymes that boost the hydrolysis of recalcitrant polysaccharides, such as chitin. They are found extensively in different insect species and are classified as auxiliary activities family 15 (AA15) LPMOs (LPMO15). Some of them were identified from the insect midgut and proven to act on chitin. However, knowledge about their physiological roles during insect growth and development remains limited. Here, we found that midgut‐specific LPMO15s are widely distributed in different insect orders, such as the orthopteran Locusta migratoria and the lepidopteran Bombyx mori. Using L. migratoria as a model insect, the function of midgut‐specific LmLPMO15‐3 during development was investigated. Double‐stranded RNA‐mediated downregulation of LmLPMO15‐3 expression at the 4th or 5th instar nymph stage severely decreased the survival rate and resulted in lethal phenotypes. Hematoxylin and eosin staining results indicated that the deficient individuals exhibited incompletely digested peritrophic matrix (PM), which suggested that LmLPMO15‐3 is essential for the deconstruction of the PM during molting. This study provides direct evidence of the physiological importance of a midgut‐specific LPMO15 during insect development. As L. migratoria is one of the most destructive agricultural pests, LmLPMO15‐3 is a potential target for pest management. [ABSTRACT FROM AUTHOR]

Paranosema locustae is an environmentally friendly parasitic predator with promising applications in locust control. In this study, transcriptome sequencing was conducted on gonadal tissues of Locusta migratoria males and females infected and uninfected with P. locustae at different developmental stages. A total of 18,635 differentially expressed genes (DEGs) were identified in female ovary tissue transcriptomes, with the highest number of DEGs observed at 1 day post-eclosion (7141). In male testis tissue transcriptomes, a total of 32,954 DEGs were identified, with the highest number observed at 9 days post-eclosion (11,245). Venn analysis revealed 25 common DEGs among female groups and 205 common DEGs among male groups. Gene ontology and Kyoto Encyclopaedia of Genes and Genome analyses indicated that DEGs were mainly enriched in basic metabolism such as amino acid metabolism, carbohydrate metabolism, lipid metabolism, and immune response processes. Protein-protein interaction analysis results indicated that L. migratoria regulates the expression of immune- and reproductive-related genes to meet the body's demands in different developmental stages after P. locustae infection. Immune- and reproductive-related genes in L. migratoria gonadal tissue were screened based on database annotation information and relevant literature. Genes such as Tsf , Hex1 , Apolp-III , Serpin , Defense , Hsp70 , Hsp90 , JHBP , JHE , JHEH1 , JHAMT , and VgR play important roles in the balance between immune response and reproduction in gonadal tissues. For transcriptome validation, Tsf , Hex1 , and ApoLp-III were selected and verified by quantitative real-time polymerase chain reaction (qRT-PCR). Correlation analysis revealed that the qRT-PCR expression patterns were consistent with the RNA-Seq results. These findings contribute to further understanding the interaction mechanisms between locusts and P. locustae .

Disruption of morphogenesis, an essential process in organismal development, can lead to disruption of biological processes, reduction in fitness, or even death of an organism. The roles of lethal giant larvae (Lgl) protein in maintaining tissue organization have been studied extensively in mammals, but little is known about this gene's roles in promoting correct tissue morphogenesis in insects. In this study, we identified an Lgl ortholog in Locusta migratoria. RT‐qPCR results revealed that LmLgl was constitutively expressed during third, fourth, and fifth instar nymphs. Furthermore, LmLgl showed highest expression in the ovary followed by wing pads, midgut, hindgut, Malpighian tubules, and foregut of the third‐instar nymphs. To examine the role of LmLgl in L. migratoria development, RNA interference was performed during nymphal stages. Silencing of LmLgl increased body size but decreased bodyweight by 9.0%. Histological sections of the midgut revealed abnormal large masses of disordered epithelial cells in dsLmLgl‐injected nymphs. In addition, downregulation of LmLgl transcript levels significantly altered the morphological structure in midgut, resulting in the formation of tumor‐like structures. Our results indicated that LmLgl may act as a tumor‐suppressor gene, which plays an essential role in maintaining a normal morphological structure in the midgut of L. migratoria. Our results also suggest that LmLgl may be explored as a potential target for developing dsRNA‐based biological pesticides for managing insect pests. [ABSTRACT FROM AUTHOR]

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Climate change favors the establishment of new pests in Germany, which now find suitable habitats here due to the changed climate. Field locusts occur repeatedly as agricultural pests in southern European countries. Therefore, it is investigated whether the climate change-induced northward shift of warmer zones can create climatically suitable habitats for field locusts in Germany and whether agricultural areas can be affected by this. The CLIMEX software is used to model the possible distribution of the Italian locust (Calliptamus italicus (L., 1758)), the Moroccan locust (Dociostaurus maroccanus (Thunberg, 1815)) and the Migratory locust (Locusta migratoria (L., 1758)) for 20 locations in Germany in six scenarios. These result from the combination of the two observation periods 2021 - 2050 and 2071 - 2100 with the three climate projections RCP2.6, RCP4.5 and RCP8.5. Based on the study, C. italicus is expected to spread widely in Germany, whereas D. maroccanus and L. migratoria might form only small and local populations. Locust swarms can potentially threaten crop products on around 10 - 25% of the agricultural area in Germany at the sites considered, but are unlikely to occur, since the intensive use of grassland areas provides insufficient conditions for reproduction. The creation of lager fallow areas as part of environmental protection and climate adaptation measures could change this in the future. In addition, swarm formation in neighbouring countries and possible migration routes to Germany should be investigated. Furthermore, the development of concepts for prevention and intervention in the event of a locust invasion is recommended. Overall, however, a low risk potential of field locusts for German agriculture is currently assumed. [ABSTRACT FROM AUTHOR]

The troponin (Tn) complex, the sensor for Ca2+ to regulate contraction of striated muscle, is composed of three subunits, that is, TnT, TnI and TnC. Different isoforms of TnI and TnC are expressed in the thorax dorsal longitudinal muscle (flight muscle) and the hind leg extensor tibiae muscle (jump muscle) of the migratory locust, Locusta migratoria. The major Tn complexes in the flight muscle and the jump muscle are Tn‐171 (TnT1/TnI7/TnC1) and Tn‐153 (TnT1/TnI5/TnC3), respectively. Here, we prepared a number of recombinant Tn complexes and the reconstituted thin filaments, and investigated their regulation on thin filament. Although both Tn‐171 and Tn‐153 regulate thin filament in a Ca2+‐dependent manner, the extent of Ca2+ activation mediated by Tn‐171 was significantly lower than that by Tn‐153. We demonstrated that TnC1 and TnC3, rather than TnI5 and TnI7, are responsible for the different levels of the thin filament activation. Mutagenesis of TnC1 and TnC3 shows that the low level of TnC1‐mediated thin filament activation can be attributed to the noncanonical residue Leu60 in the EF‐hand‐II of TnC1. We therefore propose that, in addition to Ca2+, other regulatory mechanism(s) is required for the full activation of locust flight muscle. [ABSTRACT FROM AUTHOR]

Characteristics and the use as culture media of protein hydrolysate from Locusta migratoria were determined in comparison with different peptones. After powdering, it was hydrolyzed chemically (acid hydrolysis) and obtained product Locust Peptone (LP). The contents of nitrogen, protein, fat, ash, total sugars, minerals and amino acids of LP were determined and it was seen that it has both organic and inorganic materials enough to use as a component of the medium. The effects of different concentrations added 20g/l glucose of LP on the growth of four test bacteria (Staphylococcus aureus, Bacillus subtilis, Pseudomonas putida and Escherichia coli) and test yeasts (Rhodotorula glutinis, Candida albicans, Saccharomyces cerevisiae) were investigated and it was found that the optimal concentration for bacteria and yeast are %0,6 and LP was compared with bacteriological peptone (BP), fish peptone (FP) and meat peptone (MP) .The obtained results with surface streaking and shaking culture procedures showed that LP yielded a little higher or equal FP and BP in both normal bacteria, but these values were lower than values obtained from MP. The results show that LP performed similar to or even better than commercial peptones as nitrogen sources for microorganisms growth. A new peptone has been developed from locust for microbiological media in the present study. [ABSTRACT FROM AUTHOR]

The gut microbiota of insects plays a vital role in digestion, nutrient acquisition, metabolism of dietary toxins, pathogen immunity and maintenance of gut homeostasis. Bacillus thuringinensis (Bt) poisons target insects through its toxins that are activated in the insect gut. The effects of Bt toxins on gut microbiota of insects and their underlying mechanisms are not well understood. In this study, we found that Cry1Ab/2Ab toxins significantly changed the gut bacterial community’s structure and reduced the total load of gut bacteria in the Locusta migratoria. In addition, Cry toxins significantly increased the level of reactive oxygen species (ROS) in the gut of locusts. Our results also showed that Cry1Ab/2Ab toxins induced the host gut’s immune response by up-regulating of key genes in the Immune deficiency (IMD) and Toll pathway. RNA interference showed that knocking down Relish could narrow the difference in the load, diversity, and composition in gut bacteria caused by Cry toxins. Our findings suggest that Bt potentially influences the gut bacterial community of L. migratoria through host immune response.

Energy metabolism is essential for insect development, reproduction and detoxification. Insects often reallocate energy and resources to manage external stress, balancing the demands of detoxification and reproduction. Glucose transport 4 (Glut4), a glucose transporter, is involved in glucose and lipid metabolism. However, the specific molecular mechanism of Glut4 in insect reproduction, and its role in the response to insecticide-induced oxidative stress remain unclear. In this study, LmGlut4 was identified and analyzed in Locusta migratoria. Silencing of LmGlut4 significantly reduced vitellogenin (Vg) biosynthesis in the fat body and Vg absorption by oocytes, ultimately hindering ovarian development and oocyte maturation. Knockdown of LmGlut4 also inhibited the biosynthesis of key insect hormones, such as juvenile hormone (JH), 20-hydroxyecdysone (20E) and insulin. Furthermore, LmGlut4 knockdown led to reduced triglyceride (TG) and glycogen content in the fat body and ovary, as well as decreased capacity for trehalose biosynthesis in adipocytes. Additionally, ds LmGlut4 -treated locusts showed heightened sensitivity to deltamethrin, leading to increased triglyceride depletion during detoxification. This study sheds light on the biological function of LmGlut4 in the ovary and provides potential target genes for exploring biological pest management strategies. Schematic representation of how LmGlut4 affects female reproduction in Locusta migratoria. LmGlut4 directly or indirectly induces, at least in part via the JH, 20E and insulin pathways, Vg synthesis in the fat body and utilization in developing oocyte. ds LmGlut4 disrupts triglyceride and glycogen storage in fat bodies and oocytes, leading to severe energy depletion in deltamethrin-treated locusts. Figure concept references (Wang et al., 2021; Zheng et al., 2021). [Display omitted] • LmGlut4 regulates vitellogenin (Vg) biosynthesis and oocyte maturation in Locusta migratoria. • Knockdown of LmGlut4 disrupts hormone biosynthesis (JH / 20E and insulin). • Knockdown of LmGlut4 affects glycolipid metabolism. • LmGlut4 deficiency exacerbates energy depletion due to oxidative stress induced by deltamethrin. [ABSTRACT FROM AUTHOR]

The representation of odors in the locust antennal lobe with its >2,000 glomeruli has long remained a perplexing puzzle. We employed the CRISPR-Cas9 system to generate transgenic locusts expressing the genetically encoded calcium indicator GCaMP in olfactory sensory neurons. Using two-photon functional imaging, we mapped the spatial activation patterns representing a wide range of ecologically relevant odors across all six developmental stages. Our findings reveal a functionally ring-shaped organization of the antennal lobe composed of specific glomerular clusters. This configuration establishes an odor-specific chemotopic representation by encoding different chemical classes and ecologically distinct odors in the form of glomerular rings. The ring-shaped glomerular arrangement, which we confirm by selective targeting of OR70a-expressing sensory neurons, occurs throughout development, and the odor-coding pattern within the glomerular population is consistent across developmental stages. Mechanistically, this unconventional spatial olfactory code reflects the locust-specific and multiplexed glomerular innervation pattern of the antennal lobe. [Display omitted] • A transgenic line was established to express GCaMP6s in olfactory sensory neurons • Calcium imaging reveals a functionally ring-shaped odor code of glomerular clusters • Distinct ring regions encode chemical identity and ecological significance • The ring-shaped odor-coding pattern occurs consistently throughout development Migratory locusts, genetically expressing the calcium sensor GCaMP6s in olfactory sensory neurons, have an unconventional, ring-shaped olfactory code of glomerular clusters in the antennal lobe, a pattern that occurs consistently throughout development. [ABSTRACT FROM AUTHOR]

Syntaxin5 (Syx5) belongs to SNAREs family, which play important roles in fusion of vesicles to target membranes. Most of what we know about functions of Syx5 originates from studies in fungal or vertebrate cells, how Syx5 operates during the development of insects is poorly understood. In this study, we investigated the role of LmSyx5 in the gut development of the hemimetabolous insect Locusta migratoria. LmSyx5 was expressed in many tissues, with higher levels in the gut. Knockdown of LmSyx5 by RNA interference (RNAi) considerably suppressed feeding in both nymphs and adults. The ds LmSyx5- injected locusts lost body weight and finally died at a mortality of 100%. Furthermore, hematoxylin-eosin staining indicated that the midgut is deformed in ds LmSyx5 -treated nymphs and the brush border in midgut epithelial cells is severely damaged, suggesting that LmSyx5 is involved in morphogenesis of the midgut. TEM further showed that the endoplasmic reticulum of midgut cells have a bloated appearance. Taken together, these results suggest that LmSyx5 is essential for midgut epithelial homeostsis that affects growth and development of L. migratoria. Thus, Syx5 is a promising RNAi target for controlling L. migratoria , and even other pests. [Display omitted] • Knockdown of LmSyx5 caused 100% mortality by regulating feeding. • LmSyx5 plays important roles in the development of the midgut. • The endoplasmic reticulum morphology required LmSyx5 function. [ABSTRACT FROM AUTHOR]

In this study, we shed light for the first time on the usage of migratory locusts (Locusta migratoria) as an insect model to investigate the nanotoxicological influence of aluminum oxide (Al2O3) nanoparticles at low doses on testes, and evaluate the capacity of a whole-body extract of American cockroaches (Periplaneta americana) (PAE) to attenuate Al2O3 NPs-induced toxicity. Energy dispersive X-ray microanalyzer (EDX) analysis verified the bioaccumulation of Al in testicular tissues due to its liberation from Al2O3 NPs, implying their penetration into the blood–testis barrier. Remarkably, toxicity with Al engendered disorders of antioxidant and stress biomarkers associated with substantial DNA damage and cell apoptosis. Furthermore, histopathological and ultrastructural analyses manifested significant aberrations in the testicular tissues from the group exposed to Al2O3 NPs, indicating the overproduction of reactive oxygen species (ROS). Molecular docking analysis emphasized the antioxidant capacity of some compounds derived from PAE. Thus, pretreatment with PAE counteracted the detrimental effects of Al in the testes, revealing antioxidant properties and thwarting DNA impairment and cell apoptosis. Moreover, histological and ultrastructural examinations revealed no anomalies in the testes. Overall, these findings substantiate the potential applications of PAE in preventing the testicular impairment of L. migratoria and the conceivable utilization of locusts for nanotoxicology studies.

The majority of terrestrial plants can form symbiotic associations on their roots with arbuscular mycorrhizal fungi (AMF) in the soil to stimulate the growth and nutrient uptake of the host plant and to improve plant resistance to insects and disease. However, the use of AMF for insect control on gramineous forages requires further study. Here, we evaluated the effects of AMF (Funneliformis mosseae) inoculation on the defense against Locusta migratoria attack in Elymus nutans. Inoculation assays showed that mycorrhizal plants had a higher resistance than non-inoculated plants, as evidenced by plants having more plant biomass, a higher nitrogen and phosphorus content, and greater lipoxygenase (LOX) activity. The results of insect damage showed that in addition to a decrease in the enzyme phenylalanine-ammonia-lyase, the activities of other plant defense-related enzymes (including polyphenol oxidase and β-1,3-glucanase) were increased. A key enzyme, LOX, belonging to the jasmonic acid (JA) signaling pathway was notably increased in mycorrhizal treatment. Volatile organic compounds (VOCs) were identified using gas chromatography mass spectrometry and the results showed that several metabolites with insect-resistant properties, including D-Limonene, p-Xylene, 1,3-Diethylbenzene were detected in mycorrhizal plants. These findings suggest that mycorrhizal inoculation has potential applications in insect management on forage grasses and demonstrates that the JA signaling pathway is essential for insect resistance in Elymus nutans. [ABSTRACT FROM AUTHOR]

Ecdysone‐induced protein 93F (E93) plays important roles during the metamorphosis process in insects. In this study, a cDNA of the LmE93 gene was identified from the transcriptome of Locusta migratoria, which consists of the 3378‐nucleotide open‐reading frame (ORF) and encodes 1125 amino acids with helix–turn–helix (HTH) motifs. Reverse transcription quantitative polymerase chain reaction analysis revealed that LmE93 was highest expressed in ovary. The LmE93 expression level was markedly low from the 3rd to 4th instar nymphs, and greatly increased in 1‐day‐old 5th instar nymphs with a peak on middle nymphal days, then declined in the late nymphal days. Moreover, injected dsLmE93 into 4th and 5th instar nymphs greatly reduced LmE93 transcripts, respectively, and prevented the process of metamorphosis, causing supernumerary nymphal stages. Hematoxylin–eosin staining of the integument showed that the apolysis occurred in advance in 4th instar nymphs, and old cuticle degradation was decreased in dsLmE93‐injected locusts of 5th instar nymphs. Smaller and no fully developed wings with reduced columns between the anterior and posterior regions were found in N6 and N7 supernumerary nymphs. In addition, the development of the ovary in dsLmE93‐injected locusts was severely blocked, the yolk was almost not formed and there was no development of ovarioles. The results indicated that LmE93 play key roles in the metamorphosis, cuticle, wing and ovarian development of locusts. [ABSTRACT FROM AUTHOR]