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Nanotechnology is a new trend opening up many opportunities in several fields such as agriculture and medicine. In the context of pest control and evaluation of the effect of nanomaterial on orthopteran pests, a study on the impact of silica nanoparticles on third instar larvae and adults of Locusta migratoria (Orthoptera: acrididae) was undertaken. Indeed, different concentration of nanomaterials was used in order to investigate the impact of this substance. On the other hand, neuro-intoxication symptoms were observed following ingestion of the grass sprayed with Silica nanoparticles; hyperexcitability separated by moments of immobility, movement disorders and intense defecation, testifying to the entomotoxic action of silica nanoparticles on the Migratory Locust. The mortality rate of treated locusts with the highest dose reached 100% after 11 days for L3 larvae and 21 days for adults. The evaluation of lethal times 50 (LT50) for this nanomaterial showed the speed action of this substance, which is 5.35 days for L3 larvae and 11.09 days in adult case for a maximum concentration of 5 mg/100 mL. [ABSTRACT FROM AUTHOR]

Summary: The slow lethality of fungal biopesticides to insects restrains their widespread application as a strategy of pest control. In this study, unary, binary and ternary transgenic Metarhizium robertsii were created by integrating genes that encode the scorpion neurotoxin BjαIT, the cuticle‐degrading protease Pr1A, and a double‐stranded RNA (dsRNA) that targets host gnbp3, individually or collectively under a constitutive promoter to enhance virulence. Compared with the parental wild type, all unary transgenic strains had increased virulence against four insect species, Tenebrio molitor, Locusta migratoria, Plutella xylostella and Galleria mellonella, whereas the binary transgenic strain expressing both pr1A and BjαIT had increased virulence to T. molitor and L. migratoria, with no change in virulence against P. xylostella and G. mellonella. Importantly, all ternary transgenic strains simultaneously expressing pr1A, BjαIT, and the dsRNA specific to host gnbp3 exhibited the highest increase in insect‐specific virulence. This finding highlights a novel strategy for genetic engineering of dsRNAs that target genes associated with the host immune response alongside virulence genes to maximize fungal virulence and lethality against insect pests. [ABSTRACT FROM AUTHOR]

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Food and Food Allergens (NDA) was asked to deliver an opinion on the safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a novel food pursuant to Regulation (EU) 2015/2283. The term migratory locust refers to the adult of the insect species Locusta migratoria. The NF is proposed in three formulations i) frozen without legs and wings; ii) dried without legs and wings; iii) ground with legs and wings. The main components of the NF are protein, fat and fibre (chitin) in the dried form of the NF, and water, protein, fat and fibre (chitin) in the frozen form of the NF. The Panel notes that the concentration of contaminants in the NF depends on the occurrence levels of these substances in the insect feed. The Panel notes that there are no safety concerns regarding the stability of the NF if the NF complies with the proposed specification limits during its entire shelf-life. The NF has a high protein content, although the true protein levels in the NF are overestimated when using the nitrogen-to-protein conversion factor of 6.25, due to the presence of non-protein nitrogen from chitin. The applicant proposed to use the NF as frozen, dried and ground in the form of snack, and as a food ingredient in a number of food products. The target population proposed by the applicant is the general population. The Panel notes that considering the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted history of use and toxicity studies from literature did not raise safety concerns. The Panel considers that the consumption of the NF might trigger primary sensitisation to L. migratoria proteins and may cause allergic reactions in subjects with allergy to crustaceans, mites and molluscs. Additionally, allergens from the feed may end up in the NF. The Panel concludes that the NF is safe under the proposed uses and use levels. [ABSTRACT FROM AUTHOR]

Many insect species have several genes coding for insulin-related peptides (IRPs), but so far only a single IRP gene has been identified in migratory locusts. Here, we report and characterize two other genes coding for peptides that are related to insulin, namely gonadulin and arthropod insulin-like growth factor (aIGF); peptides postulated to be orthologs of Drosophila melanogaster insulin-like peptides 8 and 6 respectively. In Locusta migratoria the aIGF transcript is expressed in multiple tissues as was previously reported for IRP in both L. migratoria and Schistocerca gregaria , but there are significant differences in expression patterns between the two species. The gonadulin transcript, however, seems specific to the ovary, whereas its putative receptor transcript is expressed most abundantly in the ovary, fat body and the central nervous system. Since the central nervous system-fat body-ovary axis is essential for successful reproduction, we studied the influence of gonadulin on vitellogenesis and oocyte growth. A reduction in the gonadulin transcript (via RNA interference) led to a significant reduction in vitellogenin mRNA levels in the fat body and a strong oocyte growth inhibition, thus suggesting an important role for gonadulin in reproduction in this species. [ABSTRACT FROM AUTHOR]

Wings are an important flight organ of insects and their morphogenesis depends on a series of cell‐to‐cell and cell‐to‐extracellular matrix interactions. Integrin as a transmembrane protein receptor mediates cell‐to‐cell adhesion, cell‐to‐extracellular matrix interactions and signal transduction. In the present study, we characterized an integrin gene that encodes integrinβ‐PS protein in Locusta migratoria. LmIntegrinβ‐PS is highly expressed in the wing pads and the middle stages of 5th instar nymphs. Immunohistochemical analysis revealed that the LmIntegrinβ‐PS protein was localized at the cell base of the two layers of wings. After suppression of LmIntegrinβ‐PS by RNA interference, the wing pads or wings were unable to form normally, with a blister wing appearance during nymph to nymph transition and nymph to adult transition. We further found that the dorsal and ventral epidermis of the wings after dsLmIntegrinβ‐PS injection were improperly connected and formed huge cavities revealed by hematoxylin and eosin staining. Furthermore, the morphology and structure of the wing cuticle was significantly disturbed which affected the stable arrangement and attachments of the wing epidermis. Moreover, the expression of related cell adhesion genes was significantly decreased in LmIntegrinβ‐PS‐suppressed L. migratoria, suggesting that LmIntegrinβ‐PS is required for the morphogenesis and development of wings during molting by stabilizing cell adhesion and maintaining the cytoskeleton of these cells. [ABSTRACT FROM AUTHOR]

BACKGROUND: Diapause plays a vital role in the survival and population reproduction of the migratory locust in temperate regions. Although some studies have suggested that photoperiod and temperature are two important environmental factors involved in the induction and termination of embryonic diapause in locusts, the independent or synergetic roles of these two factors are still unclear. RESULTS: We designed crossover experiments to investigate the effects of photoperiod, temperature and treatment time on the induction and termination of embryonic diapause in locusts by optical coherence tomography (OCT), which can track the entire embryonic developmental process via noninvasive 3D real‐time imaging. Diapause induction experiments showed that a short photoperiod (in adults) and a low egg treatment temperature are the most important prerequisites for inducing egg diapause. The diapause rate reached 90.7% when the adults were reared under a short photoperiod (8 L:16 D) and the eggs were treated at 22 °C. However, the adult rearing temperature had no significant effect on diapause induction. Analysis of variance also confirmed that there was a significant interactive effect between parental photoperiod and egg treatment temperature. Moreover, diapause termination experiments showed that 13 °C (near the developmental threshold temperature of 15 °C) for 20 days is the most effective condition for terminating diapause. CONCLUSION: Our study provides accurate photoperiod and temperature thresholds for inducing and terminating diapause in the migratory locust, suggesting potential applications in the prediction and control of locust plagues. © 2021 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Scalloped (Sd) is transcription factor that regulates cell proliferation and organ growth in the Hippo pathway. In the present research, LmSd was identified and characterized, and found to encode an N-terminal TEA domain and a C-terminal YBD domain. qRT-PCR showed that the LmSd transcription level was highest in the fifth-instar nymphs and very little was expressed in embryos. Tissue-specific analyses showed that LmSd was highly expressed in the wing. Immunohistochemistry indicated that LmSd was highly abundant in the head, prothorax, and legs during embryonic development. LmSd dsRNA injection resulted in significantly down-regulated transcription and protein expression levels compared with dsGFP injection. Gene silencing of LmSd resulted in deformed wings that were curved, wrinkled, and failed to fully expand. Approximately 40% of the nymphs had wing pads that were not able to close normally during molting from fifth-instar nymphs into adults. After silencing of LmSd , the transcription levels of cell division genes were suppressed and the expression levels of apoptosis genes were significantly up-regulated. Our results reveal that LmSd plays an important role in wing formation and development by controlling cell proliferation and inhibiting apoptosis. [ABSTRACT FROM AUTHOR]

Objective: To investigate the biochemical characterization of the carboxylesterase LmCesA1 from Locusta migratoria. Results: We expressed recombinant LmCesA1 in Sf9 cells by using the Bac-to-bac baculovirus expression system. Enzyme kinetic assays showed that the Km values of LmCesA1 for α-naphthyl acetate (α-NA) and β-naphthyl acetate (β-NA) were 0.08 ± 0.01 mM and 0.22 ± 0.03 mM, respectively, suggesting that LmCesA1 has a higher affinity for α-NA. LmCesA1 retained its enzymatic activity during incubations at pH 7–10 and at 10–30 °C. In an inhibition experiment, two organophosphate pesticides (malaoxon and malathion) and one pyrethroid pesticide (deltamethrin) showed different inhibition profiles against purified LmCesA1. Recombinant LmCesA1 activity was significantly inhibited by malaoxon in vitro. UPLC analysis showed that no metabolites were detected. Conclusions: These results suggest that overexpression of LmCesA1 enhances malathion sequestration to confer malathion tolerance in L. migratoria. [ABSTRACT FROM AUTHOR]

The desert locust, Schistocerca gregaria (Forsk.) (Orthoptera: Acrididae), avoids laying egg pods in sand containing locust fecal extracts. In this study, the oviposition inhibitory (OI) effect was investigated for a different locust species, the migratory locust Locusta migratoria (L.) (Orthoptera: Acrididae). Females of L. migratoria laid more egg pods in control sand than in sand containing fecal extracts obtained from L. migratoria and S. gregaria. However, L. migratoria oviposition was less sensitive to fecal extracts than was S. gregaria oviposition. The OI effect was observed in L. migratoria irrespective of whether the fecal extracts were prepared from gregarious or solitarious locusts, suggesting that phase-specific metabolism is not involved in OI activity. Heating and bleaching of the fecal extracts reduced the amounts of fungi and bacteria in the extracts; however, the resultant extracts still led to OI behavior in L. migratoria. These results suggest that heat-stable and bleach-stable factors in fecal extracts are responsible for the OI effect in L. migratoria. [ABSTRACT FROM AUTHOR]

7-dehydrocholesterol (7-DHC) is a key intermediate product used for biosynthesis of molting hormone. This is achieved through a series of hydroxylation reactions catalyzed by the Halloween family of cytochrome P450s. Neverland is an enzyme catalyzes the first reaction of the ecdysteroidogenic pathway, which converts dietary cholesterol into 7-DHC. However, research on the physiological function of neverland in orthopteran insects is lacking. In this study, neverland from Locusta migratoria (LmNvd) was cloned and analyzed. LmNvd was mainly expressed in the prothoracic gland and highly expressed on days 6 and 7 of fifth instar nymphs. RNAi-mediated silencing of LmNvd resulted in serious molting delays and abnormal phenotypes, which could be rescued by 7-DHC and 20-hydroxyecdysone supplementation. Hematoxylin and eosin staining results showed that RNAi-mediated silencing of LmNvd disturbed the molting process by both promoting the synthesis of new cuticle and suppressing the degradation of the old cuticle. Quantitative real-time PCR results suggested that the mRNA expression of E75 early gene and chitinase 5 gene decreased and that of chitin synthase 1 gene was markedly upregulated after knockdown of LmNvd. Our results suggest that LmNvd participates in the biosynthesis process of molting hormone, which is involved in regulating chitin synthesis and degradation in molting cycles. [Display omitted] • LmNvd was and highly expressed in the sixth to seventh days in prothoracic gland of fifth-instar nymphs. • RNA silencing of LmNvd resulted in serious molting delay and abnormal phenotypes. • Molting delay and abnormal phenotypes can be rescued by supplementary 7-DC and 20E. [ABSTRACT FROM AUTHOR]

• Locusts are an old concern and swarms need to be controlled but they travel a lot. • A model of desert locust swarm migration was developed based on real observations. • With the possibility of flying upwind, our model better reproduces the migrations. • Food intake must be considered to regulate movement dynamics. To be efficient, locust swarm control must focus on the place where eggs are laid and hopper bands may appear. But swarms travel a lot and among all the places likely to host them, there is a need to predict to which exactly they will fly. It is then essential to consider movement dynamics to anticipate any displacement that may lead to a further reproduction of locust swarms. Swarms mostly fly downwind and sometimes upwind. We designed an agent-based model to explore swarm displacements depending on the direction of the wind and the possibility for the swarms to realise upwind flights. A primary objective was to assess how upwind flights can improve the replication – and prediction – of documented migratory paths. We looked at the effects of using upwind flight on the swarm ratio arriving in expected (i.e. historically known) areas. Our simulations clearly showed that using upwind flight helped for a better replication of Schistocerca gregaria migrations than not using upwind flight. Not using upwind flight reduced swarm dispersion and reduced the range of migrations. Hence, prevailing winds alone cannot explain locust swarm migrations. Food intake must also be considered to regulate movement dynamics and vegetated areas seem to be more attractive to locusts than expected. Our simulations did not perfectly reproduce the general patterns of migrations in some scenarios, but this invites further investigations and the use of other types of field data to calibrate the model. Nonetheless, our results highlighted the importance of upwind flight and showed the major role of wind and temperature on swarm displacement. [ABSTRACT FROM AUTHOR]

This feasibility study reports the use of direct analysis in real-time high-resolution mass spectrometry (DART-HRMS) in profiling the powders from edible insects, as well as the potential for the identification of different insect species by classification modeling. The basis of this study is the revolution that has occurred in the field of analytical chemistry, with the improved capability of ambient mass spectrometry to authenticate food matrices. In this study, we applied DART-HRMS, coupled with mid-level data fusion and a learning method, to discriminate between Acheta domesticus (house cricket), Tenebrio molitor (yellow mealworm), Locusta migratoria (migratory locust), and Bombyx mori (silk moth). A distinct metabolic fingerprint was observed for each edible insect species, while the Bombyx mori fingerprint was characterized by highly abundant linolenic acid and quinic acid; palmitic and oleic acids are the statistically predominant fatty acids in black soldier fly (Hermetia illucens). Our chemometrics also revealed that the amino acid proline is a discriminant molecule in Tenebrio molitor, whereas palmitic and linoleic acids are the most informative molecular features of the house cricket (Acheta domesticus). Good separation between the four different insect species was achieved, and cross-validation gave 100% correct identification for all training samples. The performance of the random forest classifier was examined on a test set and produced excellent results, in terms of overall accuracy, sensitivity, and specificity. These results demonstrate the reliability of the DART-HRMS as a screening method in a future quality control scenario to detect complete substitution of insect powders.

Background: Non-Drosophila insects provide diverse aging types and important complementary systems for studies of aging biology. However, little attention has been paid to the special roles of non-Drosophila insects in aging research. Here, the aging-related features of the migratory locust, Locusta migratoria, were determined at the physiological, cellular, and transcriptional levels. Results: In physiological assessments, the flight performance and sperm state of locusts displayed clear aging-related decline in male adults. Transcriptional analyses demonstrated locusts have similar aging-related genes with model species. However, different from those of Drosophila and mammals, the organ-specific aging transcriptional features of locusts were characterized by intensive expression changes in flight muscle and fat body and little transcriptional changes in brain. The predominant transcriptional characteristics of flight muscle and fat body aging were changes in expression of mitochondrion-related genes and detoxification and phagocytosis genes, respectively. Cellular assessments revealed the incidence of mitochondrial abnormalities significantly increased in aged flight muscle, and apoptotic signals and nuclear abnormalities were enhanced in aged fat body but not in brain. In addition, some well-known aging genes and locust aging-related genes (i.e., IAP1, PGRP-SA, and LIPT1), whose roles in aging regulation were rarely reported, were demonstrated to affect lifespan, metabolism, and flight ability of locusts after RNAi. Conclusion: This study revealed multi-level aging signatures of locust, thus laying a foundation for further investigation of aging mechanisms in this famous insect in the future. [ABSTRACT FROM AUTHOR]

The cytokine receptor-like factor 3 (CRLF3) is an evolutionary conserved class 1 cytokine receptor present in all major eumetazoan groups. Endogenous CRLF3 ligands have not been identified and the physiological responses mediated by mammalian CRLF3 are poorly characterized. Insect CRLF3 is activated by erythropoietin (Epo) and several related molecules that protect mammalian neurons from stress-induced apoptosis. However, insects neither express Epo nor "classical" Epo receptor. Cell-protective effects of insect hemolymph have been described for several species. In this study, we explored the possibility that the endogenous CRLF3 ligand is contained in locust hemolymph. PCR analyses confirmed expression of crfl3 -transcripts in neurons and hemocytes of Locusta migratoria and Tribolium castaneum. Survival of locust hemocytes in primary cultures was significantly increased by supplementation of culture medium with locust hemolymph serum. Locust primary neuron cultures were also protected by locust hemolymph, though preceding exposure to fetal bovine serum changed the hemolymph dose-dependency of neuroprotection. Direct comparison of 10% hemolymph serum with recombinant human Epo in its optimal neuroprotective concentration revealed equivalent anti-apoptotic effects on hypoxia-exposed locust neurons. The same concentration of locust hemolymph serum also protected hypoxia-exposed T. castaneum neurons. This indicates that the neuroprotective factor in locust hemolymph is sufficiently conserved in insects to allow activation of neuroprotective receptors in different species. Locust hemolymph-induced neuroprotection in both L. migratoria and T. castaneum was abolished after RNAi-mediated suppression of crlf3 -expression. In summary, we report the presence of a conserved endogenous cytokine in locust hemolymph that activates CRLF3 and connected anti-apoptotic processes in hemocytes and neurons. Identification and characterization of the CRLF3 ligand will promote knowledge about cytokine evolution and may unravel cell-protective agents with potential clinical application. [ABSTRACT FROM AUTHOR]

To reveal the nature of locusts' visual reaction when stimulated by light, and to clarify the regulation characteristics of locusts' phototactic visual physiology and obtain good spectral light features for locusts' phototactic action, this study investigated locusts' visual spectrum response by characterizing their photoreceptive reaction to LED light using an AvaSpec fiber-optic spectrometer system. Locusts' phototactic response to spectral light was compared using this system. The results showed that locusts' visual reaction characterization presents a photo-induced vision spectrum effect and that by offsetting the main wavelength of light and the spectral peak intensity, a time-varying bio-regulation effect emerges. In addition, locusts' visual regulation ability to UV light is higher than that to violet light, whereas their reaction intensity is lower than to violet light, and the visual bio-regulatory force of locusts' visual system absorbing orange light to react sensitively becomes gradually higher than when absorbing green light as time goes on. Moreover, corresponding to nominal illumination with the same radiant energy and a visual spectrum response stimulated by UV, violet, orange, and green light, it appears that the visual spectrum window is symmetrical around 382 nm, 400 nm, 602 nm, and 530 nm, respectively, with no obvious difference between spectral amplitudes and having a time-varying incremental characteristic with amplitude peak width. This indicates that the stimulus intensity of UV, violet, orange, and green light exceeds locusts' visual tolerance, causing them to generate regulation inactivation as a visual physiological reaction, whereas the visual window response effect stimulated by UV light presents an illumination timeliness effect. Simultaneously, time-varying characteristics of locusts' bio-behavior intensity show that light intensity can make up for locusts' visual sensitivity differences at various spectral wavelengths. Presented with differential response time, photosensitive behavior intensity, and induction effect induced by orange light, time is superior for orange light, the stimulation effect caused by violet light is the strongest, and the phototactic synergy effect caused by UV light is the best. [ABSTRACT FROM AUTHOR]

Even though Paranosema locustae is widely used in China as a biological agent for controlling grasshoppers, the mortality rate is initially quite low. This study sought to determine whether the simultaneous use of P. locustae and Beauveria bassiana would be a more effective control strategy. Additionally, changes in the intestinal microbial communities of migratory locusts infected with the two pathogens were analyzed to investigate the roles of gut microbes in pathogen–host interactions. The mortality rate of locusts inoculated with B. bassiana and P. locustae simultaneously was not significantly higher than expected, but the mortality rates of locusts inoculated with B. bassiana 3, 6, and 9 days after inoculation with P. locustae were significantly higher than if their effects were additive, indicating synergism. A MiSeq analysis found that Weissella was the most common bacterium, representing 41.48% and 51.62% of the total bacteria in the mid‐ and hindguts, respectively, and the bacterial declines were greatest during dual infections with B. bassiana and P. locustae. The appropriately timed combined application of P. locustae and B. bassiana was more effective against locusts than either treatment alone. Moreover, the combined inoculation of the two pathogens changed the gut microflora of locusts, indicating the potential relevancy of their synergistic effects on locust control. [ABSTRACT FROM AUTHOR]

The objective of this study is to identify potential insect species comparing with commonly used protein sources based on efficiency of the in vitro digestibility on dry matter (DMd), organic matter (OMd) and crude protein (CPd) in broiler chickens, black‐meat chickens (Native breed) and quails. Each of gastric mucosa, pancreas and duodenal mucosa were obtained from proventriculus, pancreas and duodenum, respectively. Crude digestive enzyme extracts (CTE) were extracted from these organs to perform in vitro digestibility. Eighteen insect samples and six commonly used protein sources were served as substrates which were evaluated on DMd, OMd and CPd in triplicate for each substrate. The CTE from gastric mucosa was used to simulate proventriculus, whereas small intestine was simulation by adding the CTE from pancreas and duodenum. The large variation of chemical composition between insect meals was presented. For commonly used protein sources, animal proteins were higher on digestibility than plant proteins (p <.001). Quails represented a great potential to digest insect meals comparing other animals. Based on CPd results, there were potential insect species for broiler chickens (Achroia grisella: AG, Tenebrio molitor: TM and Musca domestica), black‐meat chickens (Patanga succincta, TM and AG) and quails (Hermetia illucens, Acheta domesticus and Locusta migratoria; p <.001). The evidences from this study suggest that these insect species contain a great potential to use as alternative protein sources promoting an animal production with sustainability. However, the in vivo experimentation must be performed to confirm in further study. [ABSTRACT FROM AUTHOR]

Unfortunately, those new tools will continue to get a workout, because the FAO has reported that regional flooding caused by the late-season cyclone Gati triggered locust swarm invasions that reached East Africa in December (Fröhlich and Wasike 2021). That they are fundamentally uncountable is a recurrent theme throughout the Old Testament, both literally (Psalms 105: 34, "He spoke, and the locusts came, young locusts without number") and figuratively (Judges 6:5, "they [the Midianites and the Amalekites] would come like locusts in number - both they and their camels could not be counted") (https://www.openbible.info/ topics/locusts). By 27 August, the editors of Nature took note, reporting that swarms "typically contain between 4 billion and 8 billion locusts, and can eat in a day the equivalent of what at least 3.5 million people would consume", that the food supply of 20 million people was at risk, and that locust research has been chronically underfunded (Nature 2020). A month later, Baskar (2020) quoted Keith Cressman, the senior locust forecasting officer for the UN FAO, as saying that the huge swarm in Kenya "would blanket the city of Paris 24 times over.". [Extracted from the article]

Outbreaks of locust plagues result from the long-term accumulation of high-density egg production. The migratory locust, Locusta migratoria, displays dramatic differences in the egg-laid number with dependence on population density, while solitarious locusts lay more eggs compared to gregarious ones. However, the regulatory mechanism for the egg-laid number difference is unclear. Herein, we confirm that oosorption plays a crucial role in the regulation of egg number through the comparison of physiological and molecular biological profiles in gregarious and solitarious locusts. We find that gregarious oocytes display a 15% higher oosorption ratio than solitarious ones. Activinβ (Actβ) is the most highly upregulated gene in the gregarious terminal oocyte (GTO) compared to solitarious terminal oocyte (STO). Meanwhile, Actβ increases sharply from the normal oocyte (N) to resorption body 1 (RB1) stage during oosorption. The knockdown of Actβ significantly reduces the oosorption ratio by 13% in gregarious locusts, resulting in an increase in the egg-laid number. Based on bioinformatic prediction and experimental verification, microRNA-34 with three isoforms can target Actβ. The microRNAs display higher expression levels in STO than those in GTO and contrasting expression patterns of Actβ from the N to RB1 transition. Overexpression of each miR-34 isoform leads to decreased Actβ levels and significantly reduces the oosorption ratio in gregarious locusts. In contrast, inhibition of the miR-34 isoforms results in increased Actβ levels and eventually elevates the oosorption ratio of solitarious locusts. Our study reports an undescribed mechanism of oosorption through miRNA targeting of a TGFβ ligand and provides new insights into the mechanism of density-dependent reproductive adaption in insects. Author summary: The continuous accumulation of high-density eggs laid by flying swarms of adults results in huge populations of flightless juveniles, which contributes to the outbreaks of locust plagues. An interesting phenomenon is that locusts have the phenotypic plasticity of reproduction. The gregarious locusts lay fewer big eggs than do solitarious phase locusts. In contrast, the solitarious phase locusts lay more small eggs compared to the gregarious locusts. We find the egg-laid number is not only regulated by the phase status of parents but also controlled by oosorption, a type of oocyte death. Further studies confirmed the phase-related ratio of oocyte death in the mother is regulated by a microRNA, which posttranscriptionally influences the expression level of a TGFβ ligand. This maternal effect on progeny size is especially critical for gregarious locusts to control the population size and maintain population fitness, and for solitarious locusts to enhance chance for gregarization and further enlargement of population size. This is the first study to reveal the molecular mechanism underlying the regulation of a microRNA-gene circuit for locust oocyte death to determine the offspring number. These findings can provide some important clues to develop potential drugs to prevent vast locust reproduction from a plague upsurge. [ABSTRACT FROM AUTHOR]

The migratory locust, Locustamigratoria (L.), a significant grasshopper species known for its ability to form large swarms and cause extensive damage to crops and vegetation, is subject to the influence of climate change. This research paper employs geographic information system (GIS) and MaxEnt ecological modelling techniques to assess the impact of climate change on the distribution patterns of L.migratoria . Occurrence data and environmental variables are collected and analysed to create predictive models for the current and future distribution of the species. The study highlights the crucial role of climate factors, particularly temperature and precipitation, in determining the locust's distribution. The MaxEnt models exhibit high-performance indicators, accurately predicting the potential habitat suitability of L.migratoria . Additionally, specific bioclimatic variables, such as mean temperature and annual precipitation, are identified as significant factors influencing the species' presence. The generated future maps indicate how this species will invade new regions especially in Europe. Such results predict the risk of this destructive species for many agriculture communities as a direct result of a warming world. The research provides valuable insights into the complex relationship between locust distribution and environmental factors, enabling the development of effective strategies for locust management and early warning systems to mitigate the impact on agriculture and ecosystems., Competing Interests: No conflict of interest to declare Disclaimer: This article is (co-)authored by any of the Editors-in-Chief, Managing Editors or their deputies in this journal., (Eslam M. Hosni, Areej A. Al-Khalaf, Mohamed G. Nasser, Sara M. ElShahed, Sara A. Alashaal.)

There is a growing appreciation that insect distribution and abundance are associated with the limits of thermal tolerance, but the physiology underlying thermal tolerance remains poorly understood. Many insects, like the migratory locust (Locusta migratoria), suffer a loss of ion and water balance leading to hyperkalaemia (high extracellular [K+]) in the cold that indirectly causes cell death. Cells can die in several ways under stress, and how they die is of critical importance to identifying and understanding the nature of thermal adaptation. Whether apoptotic or necrotic cell death pathways are responsible for low-temperature injury is unclear. Here, we use a caspase-3 specific assay to indirectly quantify apoptotic cell death in three locust tissues (muscle, nerves and midgut) following prolonged chilling and recovery from an injury-inducing cold exposure. Furthermore, we obtain matching measurements of injury, extracellular [K+] and muscle caspase-3 activity in individual locusts to gain further insight into the mechanistic nature of chilling injury. We found a significant increase in muscle caspase-3 activity, but no such increase was observed in either nervous or gut tissue from the same animals, suggesting that chill injury primarily relates to muscle cell death. Levels of chilling injury measured at the whole animal level, however, were strongly correlated with the degree of haemolymph hyperkalaemia, and not apoptosis. These results support the notion that cold-induced ion balance disruption triggers cell death but also that apoptosis is not the main form of cell damage driving low-temperature injury. [ABSTRACT FROM AUTHOR]

Conidial hydrophobins in fungal pathogens of plants, insects, and humans are required for fungal attachment and are associated with high virulence. They are believed to contribute to the pathogenesis of infection by preventing immune recognition. Here, we refute this generalisation offering a more nuanced analysis. We show that MacHYD3, a hydrophobin located on the conidial surface of the specialist entomopathogenic fungus Metarhizium acridum (narrow host range, kills only locusts and grasshoppers), activates specifically the humoral and cellular immunity of its own host insect, Locusta migratoria manilensis (Meyen) but not that of other non-host insects. When topically applied to the cuticle, purified MacHYD3 improved the resistance of locusts to both specialist and generalist fungal pathogens (wide host range) but had no effect on the fungal resistance of other insects, including Spodoptera frugiperda and Galleria mellonella. Hydrophobins extracted from the generalist fungal pathogens M. anisopliae and Beauveria bassiana had no effect on the resistance of locusts to fungal infection. Thus, the host locust has evolved to recognize the conidial hydrophobin of its specialist fungal pathogen, whereas conidial hydrophobins from generalist fungi are able to evade recognition. Our results distinguish the immunogenic potential of conidial hydrophobins between specialist and generalist fungi. [ABSTRACT FROM AUTHOR]

Regeneration after injury is accompanied by transient and lasting changes in the neuroarchitecture of the nervous system and, thus, a form of structural plasticity. In this review, we introduce the olfactory pathway of a particular insect as a convenient model to visualize neural regeneration at an anatomical level and study functional recovery at an electrophysiological level. The olfactory pathway of the locust (Locusta migratoria) is characterized by a multiglomerular innervation of the antennal lobe by olfactory receptor neurons. These olfactory afferents were axotomized by crushing the base of the antenna. The resulting degeneration and regeneration in the antennal lobe could be quantified by size measurements, dye labeling, and immunofluorescence staining of cell surface proteins implicated in axonal guidance during development. Within 3 days post lesion, the antennal lobe volume was reduced by 30% and from then onward regained size back to normal by 2 weeks post injury. The majority of regenerating olfactory receptor axons reinnervated the glomeruli of the antennal lobe. A few regenerating axons project erroneously into the mushroom body on a pathway that is normally chosen by second-order projection neurons. Based on intracellular responses of antennal lobe output neurons to odor stimulation, regenerated fibers establish functional synapses again. Following complete absence after nerve crush, responses to odor stimuli return to control level within 10–14 days. On average, regeneration of afferents, and re-established synaptic connections appear faster in younger fifth instar nymphs than in adults. The initial degeneration of olfactory receptor axons has a trans-synaptic effect on a second order brain center, leading to a transient size reduction of the mushroom body calyx. Odor-evoked oscillating field potentials, absent after nerve crush, were restored in the calyx, indicative of regenerative processes in the network architecture. We conclude that axonal regeneration in the locust olfactory system appears to be possible, precise, and fast, opening an avenue for future mechanistic studies. As a perspective of biomedical importance, the current evidence for nitric oxide/cGMP signaling as positive regulator of axon regeneration in connectives of the ventral nerve cord is considered in light of particular regeneration studies in vertebrate central nervous systems. [ABSTRACT FROM AUTHOR]

Exposure to environmental chemicals during in utero and early postnatal development can cause a wide range of neurological defects. Since current guidelines for identifying developmental neurotoxic chemicals depend on the use of large numbers of rodents in animal experiments, it has been proposed to design rapid and cost-efficient in vitro screening test batteries that are mainly based on mixed neuronal/glial cultures. However, cell culture tests do not assay correct wiring of neuronal circuits. The establishment of precise anatomical connectivity is a key event in the development of a functional brain. Here, we expose intact embryos of the locust (Locusta migratoria) in serum-free culture to test chemicals and visualize correct navigation of identified pioneer axons by fluorescence microscopy. We define separate toxicological endpoints for axonal elongation and navigation along a stereotyped pathway. To distinguish developmental neurotoxicity (DNT) from general toxicity, we quantify defects in axonal elongation and navigation in concentration–response curves and compare it to the biochemically determined viability of the embryo. The investigation of a panel of recognized DNT-positive and -negative test compounds supports a rather high predictability of this invertebrate embryo assay. Similar to the semaphorin-mediated guidance of neurites in mammalian cortex, correct axonal navigation of the locust pioneer axons relies on steering cues from members of this family of cell recognition molecules. Due to the evolutionary conserved mechanisms of neurite guidance, we suggest that our pioneer axon paradigm might provide mechanistically relevant information on the DNT potential of chemical agents on the processes of axon elongation, navigation, and fasciculation. [ABSTRACT FROM AUTHOR]

Lipids of the insect cuticle have important roles in resistance against the arid environment and invasion of foreign substances. Fatty acid synthase (FAS) is an important enzyme of the insect lipid synthesis pathway. In the present study, we identified three FAS genes from transcriptome data of the migratory locust, Locusta migratoria, based on bioinformatics analyses. Among them, two FAS genes (LmFAS1 and LmFAS3) are highly expressed in the integument of fifth instar nymphs. Suppression of LmFAS1 and LmFAS3 by RNA interference caused lethality during ecdysis or shortly after moulting. The weight of the locusts and the content of lipid droplets were reduced compared with those of the control. The results of gas chromatography–mass spectrometry analysis showed that knockdown of LmFAS3 led to a decrease of both cuticular hydrocarbons and inner hydrocarbons (CHCs and IHCs) contents, especially the content of methyl branched hydrocarbons. By contrast, knockdown of LmFAS1 only resulted in a decrease in the IHC content, but not that of CHCs. By consequence, in LmFAS1‐ and LmFAS3‐suppressed locusts, hydrocarbon deficiency reduced desiccation resistance and enhanced cuticle permeability and sensitivity to insecticides. These results indicate that LmFAS1 and LmFAS3 are essential for hydrocarbon production and cuticle permeability, which play influential roles in waterproofing the insect cuticle. [ABSTRACT FROM AUTHOR]

Under extreme environmental conditions, many insects enter a protective coma associated with a spreading depolarization (SD) of neurons and glia in the central nervous system (CNS). Recovery depends on the restoration of ion gradients by mechanisms that are not well understood. We investigated the effects of glybenclamide, an ATP-sensitive K+ (KATP) channel inhibitor, and pinacidil, a KATP activator, on the mechanisms involved in anoxic coma induction and recovery in Locusta migratoria. KATP channels allow for the efflux of K+ when activated, thereby linking cellular metabolic state to membrane potential. In intact locusts, we measured the time to enter a coma after water immersion and the time to recover the righting reflex after returning to normoxia. In semi-intact preparations, we measured the time to SD in the metathoracic ganglion after flooding the preparation with saline or exposing it to 100% N2 gas, and the time for the transperineurial potential to recover after removal of the saline or return to air. Glybenclamide decreased the time to coma induction, whereas pinacidil increased induction times. Glybenclamide also lengthened the time to recovery and decreased the rate of recovery of transperineurial potential after SD. These results were not the same as the effects of 10-2 M ouabain on N2-induced SD. We conclude that glybenclamide affects the CNS response to anoxia via inhibition of KATP channels and not an effect on the Na+/K+-ATPase. [ABSTRACT FROM AUTHOR]

The article discusses deserts locust swarms have devastated agricultural fields in east Africa, parts of the Middle East, as well as India and Pakistan and continue to spread. Topics include the young, flightless locusts congregate to gigantic marching bands of hoppers; and the locusts flew for the Red Sea and to Ethiopia.

BACKGROUND Cuticular hydrocarbons (CHCs) have a critical role in preventing desiccation and penetration of xenobiotics in insects. Previous studies have shown that cytochrome P450 subfamily 4G (CYP4G) enzymes are oxidative decarbonylases, essential for CHC biosynthesis. However, it is unclear whether there are functional differences between the two CYP4G genes in most insects. In Locusta migratoria, we identified two CYP4G genes (LmCYP4G62 and LmCYP4G102). LmCYP4G102 plays a critical role in the synthesis of CHCs, but the function of LmCYP4G62 is unknown. RESULTS: We identified, characterized, and compared two LmCYP4G genes, based on L. migratoria transcriptomic and genomic databases. RT‐qPCR showed that both were highly expressed in tissues with which oenocytes are associated, the integument and fat body. Immunostaining indicated that LmCYP4G62 and LmCYP4G102 were highly abundant in oenocytes in these tissues. However, the two enzymes had a different subcellular distribution, with LmCYP4G62 localized on the plasma membrane and LmCYP4G102 dispersed throughout the oenocyte cytoplasm, presumably on the endoplasmic reticulum. RNA interference‐mediated gene silencing against each of the two genes resulted in reduced CHC contents, in all classes for LmCYP4G102, but mostly shorter chain CHCs for LmCYP4G62. Silencing of both genes resulted in increased insecticide penetration through the cuticle, and increased locust susceptibility to desiccation and insecticides. CONCLUSION: Our studies suggest that both LmCYP4G62 and LmCYP4G102 contribute to hydrocarbon biosynthesis and play key roles in protecting locusts from water loss and insecticide penetration, but they are not fully redundant. Further, the two LmCYP4G genes might be used as new targets for insect pest management. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract Background The CRISPR/Cas9 system has been widely used to generate gene knockout/knockin models by inducing frameshift mutants in cell lines and organisms. Several recent studies have reported that such mutants can lead to in-frame exon skipping in cell lines. However, there was little research about post-transcriptional effect of CRISPR-mediated gene editing in vivo. Results We showed that frameshift indels also induced complete or stochastic exon skipping by deleting different regions to influence pre-mRNA splicing in vivo. In the migratory locust, the missing 55 bp at the boundary of intron 3 and exon 4 of an olfactory receptor gene, LmigOr35, resulted in complete exon 4 skipping, whereas the lacking 22 bp in exon 4 of LmigOr35 only resulted in stochastic exon 4 skipping. A single sgRNA induced small insertions or deletions at the boundary of intron and exon to disrupt the 3′ splicing site causing completely exon skipping, or alternatively induce small insertions or deletions in the exon to stochastic alter splicing causing the stochastic exon skipping. Conclusions These results indicated that complete or stochastic exon skipping could result from the CRISPR-mediated genome editing by deleting different regions of the gene. Although exon skipping caused by CRISPR-mediated editing was an unexpected outcome, this finding could be developed as a technology to investigate pre-mRNA splicing or to cure several human diseases caused by splicing mutations.

Comparative studies of insect genome size show that Orthoptera is a unique group of Insecta with a significantly enlarged genome. To determine a suitable internal standard for a large genome and to compare the effects of different internal standards on estimates of genome size, we used four internal standards to estimate nuclear DNA content in nine insect species with large genomes. The results showed that the combination of two internal standards, Locusta migratoria (♂1C = 6.20 pg, ♀1C = 6.60 pg) and Periplaneta americana ♂ (1C = 3.41 pg), was suitable for estimating large genome of Caelifera by flow cytometry. Using these two internal standards, we estimated the genome sizes of 17 species of Caelifera (12 genera in Acrididae, 2 genera in Pamphagidae, 1 genus in Pyrgomorphidae) using flow cytometry. Genomes ranged from 6.57 pg (Shirakiacris shirakii) to 18.64 pg (Bryodemella holdereri), the largest described in insects to date. These species showed significant genomic dimorphism based on sex: females had a 0.56 pg larger genome than males on average, which might be due to the sex chromosome determinism mechanism of X0(♂)/XX(♀). To test the results obtained by flow cytometry, we used k-mers of Illumina sequencing data to gauge the C-value of Calliptamus abbreviatus and Haplotropis brunneriana. The results of the two methods are slightly different. Genomes were estimated to be about 0.28 and 0.26 pg smaller, respectively, than the flow cytometry values. Furthermore, we also reconstructed the evolutionary relationships of these taxa and discuss the genome size evolution in a phylogenetic framework. [ABSTRACT FROM AUTHOR]

Cold exposure depolarizes cells in insects due to a reduced electrogenic ion transport and a gradual increase in extracellular K+ concentration ([K+]). Cold-induced depolarization is linked to cold injury in chillsusceptible insects, and the locust, Locusta migratoria, has been shown to improve cold tolerance following cold acclimation through depolarization resistance. Here we investigate how cold acclimation influences depolarization resistance and how this resistance relates to improved cold tolerance. To address this question, we investigated if cold acclimation affects the electrogenic transport capacity and/or the relative K+ permeability during cold exposure by measuring membrane potentials of warm- and cold-acclimated locusts in the presence and absence of ouabain (Na+-K+ pump blocker) or 4-aminopyridine (4-AP; voltage-gated K+ channel blocker). In addition, we compared the membrane lipid composition of muscle tissue from warm- and cold-acclimated locust and the abundance of a range transcripts related to ion transport and cell injury accumulation. We found that cold-acclimated locusts are depolarization resistant due to an elevated K+ permeability, facilitated by opening of 4-AP-sensitive K+ channels. In accordance, cold acclimation was associated with an increased abundance of Shaker transcripts (gene encoding 4-AP-sensitive voltage- gated K+ channels). Furthermore, we found that cold acclimation improved muscle cell viability following exposure to cold and hyperkalemia even when muscles were depolarized substantially. Thus cold acclimation confers resistance to depolarization by altering the relative ion permeability, but cold-acclimated locusts are also more tolerant to depolarization. [ABSTRACT FROM AUTHOR]

Apoptosis plays a major role in development, tissue renewal and the progression of degenerative diseases. Studies on various types of mammalian cells reported a pro-apoptotic function of acetylcholinesterase (AChE), particularly in the formation of the apoptosome and the degradation of nuclear DNA. While three AChE splice variants are present in mammals, invertebrates typically express two ache genes that code for a synaptically located protein and a protein with non-synaptic functions respectively. In order to investigate a potential contribution of AChE to apoptosis in insects, we selected the migratory locust Locusta migratoria. We established primary neuronal cultures of locust brains and characterized apoptosis progression in vitro. Dying neurons displayed typical characteristics of apoptosis, including caspase-activation, nuclear condensation and DNA fragmentation visualized by TUNEL staining. Addition of the AChE inhibitors neostigmine and territrem B reduced apoptotic cell death under normal culture conditions. Moreover, both inhibitors completely suppressed hypoxia-induced neuronal cell death. Exposure of live animals to severe hypoxia moderately increased the expression of ace-1 in locust brains in vivo. Our results indicate a previously unreported role of AChE in insect apoptosis that parallels the pro-apoptotic role in mammalian cells. This similarity adds to the list of apoptotic mechanisms shared by mammals and insects, supporting the hypothesized existence of an ancient, complex apoptosis regulatory network present in common ancestors of vertebrates and insects. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Remote Sensing is the property of Editorial Office of Journal of Remote Sensing & Science Publishing Co. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Vitellogenin (Vg) is a prerequisite for egg production and embryonic development after ovipositioning in oviparous animals. In many insects, juvenile hormone (JH) promotes fat body cell polyploidization for the massive Vg synthesis required for the maturation of multiple oocytes, but the underlying mechanisms remain poorly understood. Using the migratory locust Locusta migratoria as a model system, we report here that JH induces the dephosphorylation of Forkhead box O transcription factor (FoxO) through a signaling cascade including leucine carboxyl methyltransferase 1 (LCMT1) and protein phosphatase 2A (PP2A). JH promotes PP2A activity via LCMT1-mediated methylation, consequently triggering FoxO dephosphorylation. Dephosphorylated FoxO binds to the upstream region of two endocycle-related genes, cell-division-cycle 2 (Cdc2) and origin-recognition-complex subunit 5 (Orc5), and activates their transcription. Depletion of FoxO, Cdc2 or Orc5 results in blocked polyploidization of fat body cells, accompanied by markedly reduced Vg expression, impaired oocyte maturation and arrested ovarian development. The results suggest that JH acts via LCMT1-PP2A-FoxO to regulate Cdc2 and Orc5 expression, and to enhance ploidy of fat body cells in preparation for the large-scale Vg synthesis required for synchronous maturation of multiple eggs. [ABSTRACT FROM AUTHOR]

BACKGROUND Knickkopf (Knk) proteins play crucial roles in the formation of insect cuticle. Recent studies in the holometabolous insect red flour beetle (Tribolium castaneum) have shown that three Knk genes encoding TcKnk, TcKnk2 and TcKnk3 play different but essential roles at different developmental stages and in different tissues. However, the functions of Knk genes had not been fully explored in hemimetabolous insects such as the migratory locust Locusta migratoria. RESULTS: We identified three transcripts of LmKnk‐like genes LmKnk2 and LmKnk3 with the full‐length cDNA sequences, which were named as LmKnk2, LmKnk3‐FL and LmKnk3‐5′. These three transcripts were highly expressed before molting and mainly expressed in the integument. Among these genes, silencing only LmKnk3‐5′ by RNA interference (RNAi) caused molting defects and high mortality of the locusts. Injection of dsLmKnk3‐5′ dramatically decreased chitin content, but did not affect cuticle laminar ultra‐structures in the integument. After the knockdown of LmKnk3‐5′ transcript, lipid deposition on the cuticle surface was impeded, and locusts exhibited increased susceptibility to each of four insecticides in three different classes. However, no visible phenotypic changes were observed after LmKnk2 or LmKnk3‐FL was silenced by RNAi. CONCLUSION: We demonstrate that LmKnk3‐5′ is essential for cuticle formation in L. migratoria. This contrasts the findings that the cognate protein in T. castaneum TcKnk3‐5′ is dispensable for cuticle formation and survival. Hence, we provide some evidence that the function of Knk‐type proteins may be species‐specific. We therefore think that LmKnk3‐5′ may be a good target for the application of RNAi‐based technologies for species‐specific insect pest management. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND PPO (prophenoloxidase) cascade plays an important role in resisting invasion of entomogenous fungus. The 20‐hydroxyecdysone (20E) exerts potent effect on the innate immunity in many insects. However, whether 20E controls the PPO cascade system against fungi and the regulatory mechanism in insects remains unclear. RESULTS : In this study, both the proteome and transcriptome of Locusta migratoria were determined followed by the induction of 20E. Pattern recognition receptor GNBP‐2 (Gram‐negative binding proteins) has been identified that responded to 20E at both messenger RNA (mRNA) and protein levels. The PPO gene expression in fat body and PO (phenoloxidase) activity in plasma was found significantly induced after 20E injection and during the high‐20E developmental stage. However, when 20E signal was blocked by RNA interference (RNAi) of ecdysone receptor, the expression level of PPO and PO activity failed to be increased by 20E. Thus, 20E could not significantly induce the expression of PPO gene and PO activity after RNAi of GNBP‐2. Furthermore, 20E treatment notably enhanced the resistance of L. migratoria against Metarhizium anisopliae. Followed by of GNBP‐2 silencing, the mortality of nymphs was significantly increased under the stress of Metarhizium anisopliae, and 20E injection could not increase the resistance. CONCLUSION : The 20E regulates the PPO system to resist fungal invasion via regulating GNBP‐2 in worldwide pest L. migratoria. Our results provide insight into the mechanism of how 20E enhances the antimicrobial immunity, and will be beneficial for modification of entomogenous fungi targeting on hormones and the immune system. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Migration locust (Locusta migratoria, L.) or Kembara (Indonesian) grasshopperis one of the herbivorous insects that lay eggs on the ground. The spawning process requires the right place, humidity, and type of soil. Moist soil and suitable soil texture can accelerate the hatching of grasshoppers into nymphs. This study aims to analyze the effect of texture and soil moisture level on hatching eggs of migratory locust (Locusta migratoria, L.).This research was carried out in a semi-field laboratory in Kefamenanu, North Central Timor Regency. The study was conducted for three months, from October to December 2018. The research method used completely randomized designs and data analysis using the Tukey's HSD (honestly significant difference) 5% test. The results showed the texture of sandy clay loam with a humidity level of 60% (24.67 days) and 80% (22.33 days) can shorten the time of hatching eggs, and at 40% humidity on the texture of the clay lasts longer for 32.33 days. [ABSTRACT FROM AUTHOR]

The venom of Odontobuthus doriae contains several peptide toxins that interfere with the sodium channel function of cell membranes, some of which specifically act on the insect's sodium channel without affecting mammalian cells. In this study sodium channel toxins of Odontobuthus doriae were aligned to other closely related toxins by BLAST and ClustalW servers. Among these toxins, NaTx12 (OdTx12) showed more than 90% similarity to the most known beta excitatory toxin, AaHIT1; furthermore, our modeling studies confirmed high tertiary structure similarity of these proteins. OdTx12 was cloned and expressed in E.coli , using pET26-b and pET28-a expression vectors. Tris-tricine SDS-PAGE and Western blot analysis showed OdTx12 expression by pET28-a, only. After purification, bioactivity of the purified protein was analyzed by injection and oral administration to Locusta migratoria larvae, and toxicity to mammals was tested on mice. Injection of OdTx12 resulted in the killing of larvae with LD50 of 0.4 and 0.2 after 48 and 72 h respectively, but oral administration of OdTx12 had no significant effect on Locusta migratoria , nor did the injection to mice show any signs of toxicity. These results showed that OdTx12, as a novel β excitatory toxin can be considered as a candidate for insect control purposes. • With the advent of biological control methods and the use of synthetic pesticides has been decreased. • Scorpion venoms contain a large number of neurotoxic polypeptides. • OdTx12 shows more than 90% similarity to the most known beta excitatory toxin, AaHIT1. • β excitatory toxins can be considered as candidates for insect control purposes. [ABSTRACT FROM AUTHOR]

Apolipophorins are carrier proteins that bind lipids and mediate their transport from tissue to tissue in animals. Apolipophorin I and II (apoLp-II/I) are the major apolipophorins in insects. The implication of apoLp-II/I in cuticle lipid-barrier formation in insects has not been addressed to date. In the present study, we investigated the function of apoLp-II/I in the migratory locust Locusta migratoria (LmapoLp-II/I). During the development of fifth instar nymphs, LmapoLp-II/I transcript levels increased until mid-instar, and then decreased gradually until molting to the adult stage. We found that LmapoLp-II/I was predominately expressed in the fat body and the integument including oenocytes and epidermal cells. Immunodetection experiments revealed that LmapoLp-I mainly localized in the cytoplasm of oenocytes and epidermal cells. Silencing of LmapoLp-II/I caused molting defects in nymphs. Importantly, RNA interference against LmapoLp-II/I resulted in a significant decrease in the content of cuticle surface lipids including alkanes and methyl alkanes. Cuticular permeability was significantly enhanced in these nymphs in Eosin Y penetration assays. By consequence, desiccation resistance and insecticide tolerance of ds LmapoLp-II/I -treated locusts were reduced. Taken together, our results indicate that LmapoLp-II/I is involved in the transport and deposition of surface-cuticular lipids that are crucial for maintaining normal cuticle barrier function in L. migratoria. [ABSTRACT FROM AUTHOR]

Copyright of Natura Sloveniae: Revija za Terensko Biologijo / Journal of Field Biology is the property of Natura Sloveniae and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Continuous jumping behavior, a kind of endurance locomotion, plays important roles in insect ecological adaption and survival. However, the methods used for the efficient evaluation of insect jumping behavior remain largely lacking. Here, we developed a locomotion detection system named JumpDetector with automatic trajectory tracking and data analysis to evaluate the jumping of insects. This automated system exhibits more accurate, efficient, and adjustable performance than manual methods. By using this automatic system, we characterized a gradually declining pattern of continuous jumping behavior in 4th‐instar nymphs of the migratory locust. We found that locusts in their gregarious phase outperformed locusts in their solitary phase in the endurance jumping locomotion. Therefore, the JumpDetector could be widely used in jumping behavior and endurance locomotion measurement. [ABSTRACT FROM AUTHOR]

Adipokinetic hormones (AKHs), the neurohormones synthesized in the insect corpora cardiaca are known to mobilize lipids and carbohydrates for energy‐consuming activities including reproduction. However, both inhibitory and stimulatory effects of AKHs on insect reproduction have been reported, and the underlying mechanisms remain elusive. Using the migratory locust, Locusta migratoria, as a model system, we report here that AKHs are expressed in response to rhythmic diel change, and AKH III expression increases markedly at photophase. Diurnal injection of AKH III but not AKH I or AKH II in adult females stimulates vitellogenesis and egg development. In contrast, AKH treatment at scotophase represses female reproduction. RNA interference‐mediated knockdown of AKH receptor (AKHR) results in significantly reduced vitellogenin (Vg) expression in the fat body at photophase along with reduced Vg deposition in the ovary. AKHR knockdown also leads to decreased expression of Brummer, triacylglycerol lipase and trehalose transporter, accompanied by suppressed mobilization of triacylglycerol and trehalose. We propose that in addition to stimulating Vg expression at photophase, AKH/AKHR signalling is likely to regulate ovarian uptake of Vg via triacylglycerol mobilization and trehalose homeostasis. This study provides new insights into the understanding of AKH/AKHR signalling in the regulation of insect reproduction. [ABSTRACT FROM AUTHOR]

• We identified a BTB domain-containing protein 6 gene, LmBTBD6 , from Locusta migratoria transcriptome database. • LmBTBD6 was highly expressed in the wing pads and regulated by the LmEcR-LmHR39-mediated 20E signaling pathway. • Depletion of LmBTBD6 transcripts caused wing defects in the transition from nymph to adult. • LmBTBD6 regulated the expression of wing development-related genes and wing-specific cuticle protein genes. The development of insect wings is a complex process controlled by a series of genes, whereas the mechanism of wing development of orthoptera insects is less frequently reported. In the present study, a BTB domain-containing protein 6 (LmBTBD6) gene was identified from Locusta migratoria. Its encoded protein belongs to the BTB-BACK-PHR subfamily, and is highly conserved among insect species. LmBTBD6 was mainly expressed in the wing pads and showed high expression on day 7 of fifth-instar nymphs. LmBTBD6 responded to induction by 20-Hydroxyecdysone (20E) in vivo , and its expression was significantly suppressed after knocking down the ecdysone receptor gene LmEcR and nuclear receptor gene LmHR39. Deficiency of LmBTBD6 did not show visible phenotype in the wing pads transition from nymph to nymph of L. migratoria , but caused wing defects in the transition from nymph to adult. After silencing of LmBTBD6 , the transcription of wing development-related genes (LmSal411 , LmSal468 , and LmHth) and the wing-specific cuticle protein genes (LmACP7 and LmACP8) of L. migratoria were significantly suppressed. Thus, LmBTBD6 that regulated by the LmEcR-LmHR39-mediated 20E signaling pathway is involved in wing development during the nymph to adult transition by regulating the expression of wing development-related genes and wing-specific cuticle protein genes. [ABSTRACT FROM AUTHOR]

Diapause is a state of arrested growth, which allows insects to adapt to diverse environments. Serine protease inhibitors (serpins) play an important role in various physiological processes, including blood coagulation, fibrinolysis, development, complement activation and extracellular matrix remodeling. We hypothesized that serpin may affect energy metabolism and thereby control diapause of migratory locust (Locusta migratoria) embryos by regulating protease cascades. A total of seven nonredundant serpin genes (named serpin1–serpin7) of L. migratoria were obtained through transcriptomic analysis. We further performed label‐free proteomic sequencing and analysis of diapause and nondiapause eggs of L. migratoria, revealing significant differences in serpin7 expression. A significant reduction in diapause rate under the short photoperiod was observed in insects treated with serpin7 double‐stranded RNA. Furthermore, knockdown of the serpin7 gene resulted in significant upregulation of the activity of polyphenol oxidase. We therefore propose that the observed serpin7 gene plays a crucial role in diapause, suggesting that control of energy metabolism may have potential as a future strategy for the reproductive control of insect pests. [ABSTRACT FROM AUTHOR]

Animals often exhibit dramatically behavioral plasticity depending on their internal physiological state, yet little is known about the underlying molecular mechanisms. The migratory locust, Locusta migratoria, provides an excellent model for addressing these questions because of their famous phase polyphenism involving remarkably behavioral plasticity between gregarious and solitarious phases. Here, we report that a major insect hormone, juvenile hormone, is involved in the regulation of this behavioral plasticity related to phase change by influencing the expression levels of olfactory-related genes in the migratory locust. We found that the treatment of juvenile hormone analog, methoprene, can significantly shift the olfactory responses of gregarious nymphs from attraction to repulsion to the volatiles released by gregarious nymphs. In contrast, the repulsion behavior of solitarious nymphs significantly decreased when they were treated with precocene or injected with double-stranded RNA of JHAMT, a juvenile hormone acid O-methyltransferase. Further, JH receptor Met or JH-response gene Kr-h1 knockdown phenocopied the JH-deprivation effects on olfactory behavior. RNA-seq analysis identified 122 differentially expressed genes in antennae after methoprene application on gregarious nymphs. Interestingly, several olfactory-related genes were especially enriched, including takeout (TO) and chemosensory protein (CSP) which have key roles in behavioral phase change of locusts. Furthermore, methoprene application and Met or Kr-h1 knockdown resulted in simultaneous changes of both TO1 and CSP3 expression to reverse pattern, which mediated the transition between repulsion and attraction responses to gregarious volatiles. Our results suggest the regulatory roles of a pleiotropic hormone in locust behavioral plasticity through modulating gene expression in the peripheral olfactory system. Author summary: A behavioral change from shy solitarious individuals to highly social gregarious individuals is critical to the formation of disastrous swarms of locusts. However, the underlying molecular mechanism of behavioral plasticity regulated by hormones is still largely unknown. Here, we investigated the effect of juvenile hormone (JH) on the behavioral transition in fourth-instar gregarious and solitarious locusts. We found that JH induced the behavioral shift of the gregarious locust from attraction to repulsion to the volatiles of gregarious locusts. The solitarious locust significantly decreased repulsion behavior after deprivation of JH by precocene or knockdown of JHAMT, a key enzyme to synthesize JH. JH application on gregarious locusts caused significant expression alteration of genes, especially the olfactory genes TO and CSP in the antennae. We further demonstrated that the JH signaling pathway suppressed aggregation behavior in gregarious locusts by increasing TO1 expression and decreasing CSP3 expression at the same time. Our results suggested that internal physiological factors can directly modulate periphery olfactory system to produce behavioral plasticity. [ABSTRACT FROM AUTHOR]

Some long noncoding RNAs (lncRNAs) are specifically expressed in brain cells, implying their neural and behavioural functions. However, how lncRNAs contribute to neural regulatory networks governing the precise behaviour of animals is less explored. Here, we report the regulatory mechanism of the nuclear-enriched lncRNA PAHAL for dopamine biosynthesis and behavioural adjustment in migratory locusts (Locusta migratoria), a species with extreme behavioral plasticity. PAHAL is transcribed from the sense (coding) strand of the gene encoding phenylalanine hydroxylase (PAH), which is responsible for the synthesis of dopamine from phenylalanine. PAHAL positively regulates PAH expression resulting in dopamine production in the brain. In addition, PAHAL modulates locust behavioral aggregation in a population density-dependent manner. Mechanistically, PAHAL mediates PAH transcriptional activation by recruiting serine/arginine-rich splicing factor 2 (SRSF2), a transcription/splicing factor, to the PAH proximal promoter. The co-activation effect of PAHAL requires the interaction of the PAHAL/SRSF2 complex with the promoter-associated nascent RNA of PAH. Thus, the data support a model of feedback modulation of animal behavioural plasticity by an lncRNA. In this model, the lncRNA mediates neurotransmitter metabolism through orchestrating a local transcriptional loop. Author summary: The neurotransmitter dopamine is crucial for the neuronal and behavioral response in animals. Phenylalanine hydroxylase (PAH) is involved in dopamine biosynthesis and behavioral regulation in the migratory locust. However, the molecular mechanism for the fine tuning of PAH expression in behavioral response remains ambiguous. Here we discovered a nuclear-enriched lncRNA PAHAL that is transcribed from the coding strand of the PAH gene in the locust (i.e., sense lncRNA). PAHAL positively regulated PAH expression and dopamine production in the brain. In addition, PAHAL modulated behavioral aggregation of the locust. Mechanistically, PAHAL mediated the transcriptional activation of PAH by recruiting SRSF2, a transcription/splicing factor, to the promoter-associated nascent RNA of PAH. These data support a model of feedback modulation of dopamine biosynthesis and behavioral plasticity via a sense lncRNA in the catecholamine metabolic pathway. [ABSTRACT FROM AUTHOR]