Gregarious Locusts Research Articles

Structural basis of selective beta-carotene binding by a soluble protein

β-carotene (BCR) is the most abundant carotenoid, a colorant, antioxidant, and provitamin A. The extreme hydrophobicity of this hydrocarbon requires special mechanisms for distribution in aqueous media, including water-soluble carotenoproteins. However, all known carotenoproteins prefer oxygenated carotenoids and bind BCR inefficiently. Here, we present the crystal structure of the BCR-binding protein (BBP) from gregarious male locusts, which is responsible for their vivid yellow body coloration, in complex with its natural ligand, BCR. BBP forms an antiparallel tubular homodimer with α/β-wrap folded monomers, each forming a hydrophobic 47Å long, coaxial tunnel that opens outward and is occupied by one s-cisC6-C7, all-trans BCR molecule. In the BCR absence, BBP accepts a range of xanthophylls, with reduced efficiency depending on the position and number of oxygen atoms, but rejects lycopene. The structure captures a pigment complex with a Takeout 1 protein and inspires potential applications of BBP as a BCR solubilizer.

Phenotypic Plasticity in Locusts: Trade-Off Between Migration and Reproduction.

Locusts exhibit phenotypic plasticity in response to population density changes, with distinct phenotypes in the solitary and gregarious phases. In the past decade, many studies have revealed the molecular mechanisms underlying phase changes, which include the change of body coloration, pheromones, behavior, flight, fecundity, immunity, and aging. Our understanding of the molecular mechanisms related to these phenotypic differences has expanded in breadth and depth with the decoding of the locust genome, involving transcriptional, post-transcriptional, translational, and epigenetic regulation. Large-scale regulation networks composed of genes and noncoding RNAs reflect the systematic modifications of the locust phase transition in response to environmental changes. Gene manipulation techniques have verified the functions of specific genes and related pathways in phase changes. This review highlights the latest advances in studies of locust phase changes and suggests that the divergence of energy and metabolism allocation in gregarious and solitary locusts is an adaptive strategy for long-distance migration and local reproduction, respectively. Finally, we propose future research directions and discuss emerging questions in the area of phenotypic plasticity of locusts.

Dibutyl phthalate released by solitary female locusts mediates sexual communication at low density

Sex pheromones play a crucial role in mate location and reproductive success. Insects face challenges in finding mates in low-density environments. The population dynamics of locusts vary greatly, ranging from solitary individuals to high-density swarms, leading to multiple-trait divergence between solitary and gregarious phases. However, differences in sexual communication between solitary and gregarious locusts have not been sufficiently explored. Herein, we found that solitary locusts but not gregarious ones heavily rely on a single compound, dibutyl phthalate (DBP), for sexual communication. DBP is abundantly released by solitary female locusts and elicits strong attraction of male solitary and gregarious locusts. Solitary adult males display much higher electrophysiological responses to DBP than adult females. Additionally, LmigOr13 was identified as the DBP-specific odorant receptor expressed in neurons housed in basiconic sensilla. Male LmigOr13-/- mutants generated by CRISPR/Cas9 have low electrophysiological responses and behavioral attraction to DBP in both laboratory and field cage experiments. Notably, the attractiveness of DBP to male locusts becomes more evident at lower population densities imposed by controlling the cage size. This finding sheds light on the utilization of a sex pheromone to promote reproductive success in extremely low-density conditions and provides important insights into alternative approaches for population monitoring of locusts.

Insights into the molecular mechanism of yellow cuticle coloration by a chitin-binding carotenoprotein in gregarious locusts

Carotenoids are hydrophobic pigments binding to diverse carotenoproteins, many of which remain unexplored. Focusing on yellow gregarious locusts accumulating cuticular carotenoids, here we use engineered Escherichia coli cells to reconstitute a functional water-soluble β-carotene-binding protein, BBP. HPLC and Raman spectroscopy confirmed that recombinant BBP avidly binds β-carotene, inducing the unusual vibronic structure of its absorbance spectrum, just like native BBP extracted from the locust cuticles. Bound to recombinant BBP, β-carotene exhibits pronounced circular dichroism and allows BBP to withstand heating (T0.5 = 68 °C), detergents and pH variations. Using bacteria producing distinct xanthophylls we demonstrate that, while β-carotene is the preferred carotenoid, BBP can also extract from membranes ketocarotenoids and, very poorly, hydroxycarotenoids. We show that BBP-carotenoid complex reversibly binds to chitin, but not to chitosan, implying the role for chitin acetyl groups in cuticular BBP deposition. Reconstructing such locust coloration mechanism in vitro paves the way for structural studies and BBP applications.

De Novo Biosynthesis of 4-Vinylanisole in Engineered Escherichia coli.

4-Vinylanisole is an aggregation pheromone of the locust. Both gregarious and solitary locusts exhibit a strong attraction toward 4-vinylanisole, irrespective of gender or age. Therefore, 4-vinylanisole can be used for trapping and monitoring locusts. In this study, the construction of a de novo 4-vinylanisole pathway in Escherichia coli has been demonstrated for the first time. Subsequently, by increasing the supply of precursor substrates, we further improved the biosynthesis of 4-vinylanisole. Finally, a two-phase organic overlay culture was used to increase the titer to 206 mg/L. It presents a sustainable and ecofriendly alternative for the synthesis of 4-vinylanisole.

The Multivariate Regression Models Suggested as Standardising Tools for Categorising Solitarious and Gregarious Groups of the Main Pest Locust, Schistocerca gregaria, Produce Reproducible Results.

Outbreaks of the desert locust Schistocerca gregaria affect some of the poorest parts of Africa, with devastating outcomes. The key to understanding and dealing with this problematic adaptation to environmental changes is comparing gregarious and solitarious locusts, either in nature or in laboratories. Categorising locusts and detecting changes in their phase status is key to such comparisons, which have been hitherto based on applying mathematical models that use behavioural parameters and that each laboratory has to build anew for each experiment. All the models used thus far are different from one another. This implies differences in the tools used for the different experiments and by the different laboratories and, thus, potential noise in the results and interpretations. Standardising the way locusts are categorised is necessary if we want to reduce noise and errors. It is crucial if we seek to make the results and interpretations transferable and comparable between experiments and laboratories for such an important research area. To tackle this problem, we suggested two models as possible standardising tools. However, the problem of a lack of standardised tools re-emerged due to the doubts cast on the validity of those models. Here, we use samples from independent S. gregaria populations in order to test and validate those models. We discuss how successful the two models were at categorising solitarious, intermediate (transient), and gregarious nymph and adult S. gregaria samples. We highlight shortcomings and make more specific recommendations on the use of those models based on the precision differences they show when categorising solitarious and gregarious S. gregaria nymph and adult samples. Overall, both models have proven to be valid since their results were largely replicated and seem reproducible.

Robot-Locust Social Information Transfer Occurs in Predator Avoidance Contexts

Social learning is an evolutionarily important ability increasingly attributed also to invertebrate species. Interfacing robots with animals represents a promising strategy to investigate social learning. Herein, we studied if the gregarious form of Locusta migratoria, a particularly suited model to examine social learning, can use social information provided by robotic demonstrators to optimize their predator avoidance. Robotic demonstrators with different silhouettes and colours (biomimetic or neutral) were used to investigate if their rotation on a rod (e.g. hiding behaviour) elicited the same behaviour in neighbouring locusts. Locusts’ responses were affected by different robotic demonstrators, observing a significant impact of the biomimetic silhouette in reducing the latency duration, and in promoting social learning (e.g. locusts displaying hiding behaviour after observing it in robotic demonstrators). A significant impact of colour patterns in triggering socially induced hiding behaviour was also recorded, especially when the biomimetic silhouette was coloured with the gregarious-like pattern. This research indicates gregarious locusts exploit social information in specific ecological contexts, providing basic knowledge on the complex behavioural ecology and social biology in invertebrates. The proposed animal-robot interaction paradigm shows the role of robots as carrier of social information to living organisms, suggesting social biorobotics as advanced and sustainable approach for socio-biology investigation, and environmental management.

4-Vinylanisole promotes conspecific interaction and acquisition of gregarious behavior in the migratory locust

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.

Spatially differential regulation of ATF2 phosphorylation contributes to warning coloration of gregarious locusts.

Warning coloration are common defense strategies used by animals to deter predators. Pestilential gregarious locusts exhibit a notable black-brown pattern as a form of warning coloration. However, the mechanisms regulating this distinctive pattern remain largely unknown. Here, we revealed that the black and brown integuments of locusts are governed by varying amounts of β-carotene and β-carotene-binding protein (βCBP) complexes. βCBP expression is regulated by the bZIP transcription factor activation transcription factor 2 (ATF2), which is activated by protein kinase C alpha in response to crowding. Specifically, ATF2 is phosphorylated at Ser327 and translocates to the nucleus, where it binds to the βCBP promoter and stimulates overexpression. Differential phosphorylation of ATF2 leads to the divergent black and brown coloration in gregarious locusts. The accumulation of red pigments vital for creating the brown sternum depends on βCBP overexpression. The spatial variation in ATF2 phosphorylation enables locusts to rapidly adapt to changing environment for aposematism.

Integrated Management Practices against Desert Locust, Schistocerca gregaria (Forskal) in India: A Review

Locusts are the most dangerous agricultural pests. They are belonging to family Acrididae. Gregarious locusts travel in swarms from one location to another in adult form. The desert locust, Schistocerca gregaria (Forskal), is one of the grasshopper species that cause crop damage and can fly up to 150 km in the direction of the wind. The present state of knowledge on its biological regulation employing microbes and plant extracts is discussed. Metarhizium flavoviride was among the first fungi to be identified in the laboratory and field as a bio-control agent against desert locust. Following extensive investigation, with integrated pest management stratedies using these bio-controls would be a viable option for controlling desert locust infestations. Against the desert locust, IPM (Integrated Pest Management) approaches that emphasize the successful combination of chemical and biological insecticides with prediction and monitoring technology have been encouraged. Recent experimental investigations and researches are mainly focusing on identifying viable answers through financial collaboration between governmental and non-governmental organizations. The authors highlighted the loss in the agricultural sector due to desert locust infestation, as well as its sophisticated control and management solutions, after evaluating publications from numerous journals, magazines, and symposia.

PLIN2-induced ectopic lipid accumulation promotes muscle ageing in gregarious locusts.

Ageing plasticity represents the flexibility of the ageing process in response to non-genetic factors, occurring commonly in animals. However, the regulatory mechanisms underlying ageing plasticity are largely unclear. The density-dependent polyphenism of locusts, Locusta migratoria, displays dramatic lifespan divergence between solitary and gregarious phases, providing a useful system for studying ageing plasticity. Here, we found that gregarious locusts displayed faster locomotor deficits and increased muscle degeneration on ageing than solitary locusts. Comparative transcriptome analysis in flight muscles revealed significant differences in transcriptional patterns on ageing between two phases. RNA interference screening showed that the knockdown of the upregulated PLIN2 gene significantly relieved the ageing-related flight impairments in gregarious locusts. Mechanistically, the gradual upregulation of PLIN2 could induce the accumulation of ectopic lipid droplets and triacylglycerols in flight muscles during the ageing process. Further experiments suggested that ectopic lipid accumulation led to an ageing-related β-oxidation decline through limiting fatty acid transport and content. These findings reveal the key roles of lipid metabolism in the differences of muscle ageing between solitary and gregarious locusts and provide a potential mechanism underlying environment-induced muscle ageing plasticity.

Translational strategy for phase change between gregarious and solitary locusts

Translational strategy for phase change between gregarious and solitary locusts

How molting locusts avoid cannibalism

Abstract Group living has various benefits, but it also carries costs, such as risk of cannibalism. Molting is a vulnerable period of being cannibalized in juvenile arthropods, but how gregarious arthropods avoid this threat is poorly understood. Here, we examined how actively migrating gregarious nymphs of desert locust, Schistocerca gregaria, avoid cannibalism during molting, in the Sahara Desert of Mauritania. In the field, gregarious nymphs cyclically march and feed on grass during the day. Our field observations found that marching behavior helped separating pre-molting and cannibalistic non-molting nymphs. Cannibalistic non-molting nymphs marched away from roost plants, leaving sedentary pre-molting nymphs behind, creating cannibal-free spaces. Some non-molting nymphs reached a pre-molting state after daytime marching, thus both pre- and non-molting nymphs roosted on same plants at night. However, pre-molting nymphs moved away from conspecifics prior to molting. Starvation experiments confirmed that food-satiation decreased cannibalistic necrophagy. Physiological surveys of diel feeding and molting patterns revealed that nymphs molted at times when conspecifics were food-satiated rather than hungry. Hence, our results indicate that behavioral and physiological traits of gregarious locusts could function to spatiotemporally separate molting locusts from cannibalistic conspecifics, thus reducing molting-associated cannibalism. This is the first report of migration-dependent molting synchrony as a mechanism reducing costs of aggregation in gregarious arthropods.

Information integration for decision-making in desert locusts

Locust swarms can extend over hundred kilometers, and starvation compels this ancient pest to devour everything in its path. Theory suggests that gregarious behavior benefits foraging efficiency, yet the role of social cohesion remains elusive. To this end, we collected high-resolution trajectories of individual and grouped gregarious desert locusts in a 2-choice assay with patches of either similar or different quality. Carefully maintaining animals' identities allowed us to monitor each individual's experience and estimate the leaky accumulation process of personally acquired and socially derived evidence. We fitted data to a Bayesian model to gain insight into the decision-making system for patch selection. By disentangling the relative contribution of each information class, our study suggests that locusts balance incongruent evidence but reinforce congruent ones. We provide insight into the collective foraging decisions of social (but non-eusocial) insects and present locusts as a powerful empirical system to study individual choices and consequent collective dynamics.

Cytosolic and mitochondrial ribosomal proteins mediate the locust phase transition via divergence of translational profiles

The phase transition from solitary to gregarious locusts is crucial in outbreaks of locust plague, which threaten agricultural yield and food security. Research on the regulatory mechanisms of phase transition in locusts has focused primarily on the transcriptional or posttranslational level. However, the translational regulation of phase transition is unexplored. Here, we show a phase-dependent pattern at the translation level, which exhibits different polysome profiles between gregarious and solitary locusts. The gregarious locusts exhibit significant increases in 60S and polyribosomes, while solitary locusts possess higher peaks of the monoribosome and a specific "halfmer." The polysome profiles, a molecular phenotype, respond to changes in population density. In gregarious locusts, ten genes involved in the cytosolic ribosome pathway exhibited increased translational efficiency (TE). In solitary locusts, five genes from the mitochondrial ribosome pathway displayed increased TE. The high expression of large ribosomal protein 7 at the translational level promotes accumulation of the free 60S ribosomal subunit in gregarious locusts, while solitary locusts employ mitochondrial small ribosomal protein 18cto induce the assembly of mitochondrial ribosomes, causing divergence of the translational profiles and behavioral transition. This study reveals the translational regulatory mechanism of locust phase transition, in which the locusts employ divergent ribosome pathways to cope with changes in population density.

Glutamate–GABA imbalance mediated by miR-8-5p and its STTM regulates phase-related behavior of locusts

The aggregation of locusts from solitary to gregarious phases is crucial for the formation of devastating locust plagues. Locust management requires research on the prevention of aggregation or alternative and greener solutions to replace insecticide use, and insect-derived microRNAs (miRNAs) show the potential for application in pest control. Here, we performed a genome-wide screen of the differential expression of miRNAs between solitary and gregarious locusts and showed that miR-8-5p controls the γ-aminobutyric acid (GABA)/glutamate functional balance by directly targeting glutamate decarboxylase (Gad). Blocking glutamate-GABA neurotransmission by miR-8-5p overexpression or Gad RNAi in solitary locusts decreased GABA production, resulting in locust aggregation behavior. Conversely, activating this pathway by miR-8-5p knockdown in gregarious locusts induced GABA production to eliminate aggregation behavior. Further results demonstrated that ionotropic glutamate/GABA receptors tuned glutamate/GABA to trigger/hamper the aggregation behavior of locusts. Finally, we successfully established a transgenic rice line expressing the miR-8-5p inhibitor by short tandem target mimic (STTM). When locusts fed on transgenic rice plants, Gad transcript levels in the brain increased greatly, and aggregation behavior was lost. This study provided insights into different regulatory pathways in the phase change of locusts and a potential control approach through behavioral regulation in insect pests.

Faeces’ odours attract gregarious locust hoppers

Collective motion is one of the most impressive common features of gregarious locusts: once formed, bands and swarms get moving for long distances. It was shown that visual perception of neighbours plays a key role in maintaining marching behaviour at a local scale. But at a larger scale, mechanisms underlying band cohesion are less understood. It was shown in several field studies that individuals separated from the band were able to get back to the group, even after being separated since a night. In this context, faeces’ odours could be a possible indicator of the recent passage of a group. In this study, we tested if nymphs are attracted by faeces’ odours and if this effect is modulated by the age of the faeces. To this end, we conducted individual olfactometric behavioural assays of 3rd instar hoppers of desert locust, Schistocerca gregaria, exposed to odours of 1 h-old and 24 h-old faeces. We also used Gas Chromatography-Mass Spectrometry (GC–MS) to identify odours’ volatile organic compounds from faeces. The results of behavioural assays indicated a strong attractive effect of faeces, with no preference for one of the two faecal age classes. Nymphs spent significantly more time in the side of the olfactometer where the faeces’ odours came from, and 72.7% of tested individuals chose this side first. We filtered and annotated 11 volatile organic compounds present in both fresh and old faeces in GC–MS analyses, including guaiacol and phenol, which are known to cause an aggregative effect on desert locusts. As the attractive effect lasted over 24 h, band’s faeces could still have an attractive effect when individuals are separated from the band since one day. In this situation, latecomers individuals would be able to get back to the group by following the traces of their predecessors.

Climate change expected to improve digestive rate and trigger range expansion in outbreaking locusts

AbstractGlobal climate change will probably exacerbate crop losses from insect pests, reducing agricultural production, and threatening food security. To predict where crop losses will occur, scientists have mainly used correlative models of species' distributions, but such models are unreliable when extrapolated to future environments. To minimize extrapolation, we developed mechanistic and hybrid models that explicitly capture range‐limiting processes, and we explored how incorporating mechanisms altered the projected impacts of climate change for an agricultural pest, the South American locust (Schistocerca cancellata). Because locusts are generalist herbivores surrounded by food, their population growth may be limited by thermal effects on digestion more than food availability. To incorporate this mechanism into a distribution model, we measured the thermal effects on the consumption and defecation of field‐captured locusts and used these data to model energy gain in current and future climates. We then created hybrid models by using outputs of the mechanistic model as predictor variables in correlative models, estimating the potential distribution of gregarious outbreaking locusts based on multiple predictor sets, modeling algorithms, and climate scenarios. Based on the mechanistic model, locusts can assimilate relatively high amounts of energy throughout temperate and tropical South America; however, correlative and hybrid modeling revealed that most tropical areas are unsuitable for locusts. When estimating current distributions, the top‐ranked model was always the one fit with mechanistic predictors (i.e., the hybrid model). When projected to future climates, top‐ranked hybrid models projected range expansions that were 23%–30% points smaller than those projected by correlative models. Therefore, a combination of the correlative and mechanistic approaches bracketed the potential outcomes of climate change and enhanced confidence where model projections agreed. Because all models projected a poleward range expansion under climate change, agriculturists should consider enhanced monitoring and the management of locusts near the southern margin of the range.

PiRNA-guided intron removal from pre-mRNAs regulates density-dependent reproductive strategy.

Animal density-dependent experiences have profound effects on reproductive strategies with marked fecundity differences. Migratory locust adopts distinct population density-dependent reproductive strategies to cope with their respective life cycles, but the mechanisms remain poorly understood. Here, we report that Piwi-interacting RNAs (piRNAs) in the locust germline play key roles in this process. We find that the locust Piwi protein Liwi1 and piRNAs are highly expressed in early developing egg chambers in solitarious locusts, which have higher fecundity than gregarious locusts. Approximately 40% of solitarious locust-associated piRNAs map to protein-coding genes. We find that Liwi1/piRNAs facilitate pre-mRNA splicing of oocyte development-related genes, such as oo18 RNA-binding protein (Orb), in the germline by recruiting the splicing factor U2AF35 to piRNA-targeted introns, thereby increasing fecundity. Such piRNA-guided pre-mRNA splicing is also functional in Drosophila and mouse germ cells. We uncover a piRNA-guidedsplicing mechanism for processing reproduction-related mRNAs and determining animal reproductive strategies.

Aggregation pheromone 4-vinylanisole promotes the synchrony of sexual maturation in female locusts.

Reproductive synchrony generally occurs in various group-living animals. However, the underlying mechanisms remain largely unexplored. The migratory locust, Locusta migratoria, a worldwide agricultural pest species, displays synchronous maturation and oviposition when forms huge swarm. The reproductive synchrony among group members is critical for the maintenance of locust swarms and population density of next generation. Here, we showed that gregarious female locusts displayed more synchronous sexual maturation and oviposition than solitarious females and olfactory deficiency mutants. Only the presence of gregarious male adults can stimulate sexual maturation synchrony of female adults. Of the volatiles emitted abundantly by gregarious male adults, the aggregation pheromone, 4-vinylanisole, was identified to play key role in inducing female sexual maturation synchrony. This maturation-accelerating effect of 4-vinylanisole disappeared in the females of Or35-/- lines, the mutants of 4-vinylanisole receptor. Interestingly, 4-vinylanisole displayed a time window action by which mainly accelerates oocyte maturation of young females aged at middle developmental stages (3-4 days post adult eclosion). We further revealed that juvenile hormone/vitellogenin pathway mediated female sexual maturation triggered by 4-vinylanisole. Our results highlight a 'catch-up' strategy by which gregarious females synchronize their oocyte maturation and oviposition by time-dependent endocrinal response to 4-vinylanisole, and provide insight into reproductive synchrony induced by olfactory signal released by heterosexual conspecifics in a given group.