Gregarious Phase Research Articles

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.

Beyond adult models: Tribolium castaneum larval timekeeping reveals unexpected robustness and insights into circadian clock.

Circadian rhythms are self-sustained endogenous oscillations that are found in all living organisms. In insects, circadian rhythms control a wide variety of behavioral and physiological processes, including feeding, locomotion, mating, and metabolism. While the role of circadian rhythms in adult insects is well-understood, it is largely unexplored in larvae. This study investigates the potential for larval synchronized activity in the red flour beetle (Tribolium castaneum), a species exhibiting solitary and aggregation phases. We hypothesized that, similar to adults, larvae would exhibit a daily activity pattern governed by an endogenous circadian clock. We further predicted that the transition between the solitary and gregarious phases extends to unique temporal activity patterns. Our results revealed unique timekeeper gene expression in larvae, leading to a distinct daily rhythm characterized by nocturnal activity. Cues indicating on potential cannibalism did not change daily activity peak. However, the absence of these cues significantly reduced the proportion of rhythmic larvae and led to higher variation in peak activity, highlighting the crucial role of social interactions in shaping their rhythmicity. This study sheds light on the evolution and function of larval synchronization in group-living insects, offering novel insights into this complex behavior.

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.

Synergistic olfactory processing for social plasticity in desert locusts

Desert locust plagues threaten the food security of millions. Central to their formation is crowding-induced plasticity, with social phenotypes changing from cryptic (solitarious) to swarming (gregarious). Here, we elucidate the implications of this transition on foraging decisions and corresponding neural circuits. We use behavioral experiments and Bayesian modeling to decompose the multi-modal facets of foraging, revealing olfactory social cues as critical. To this end, we investigate how corresponding odors are encoded in the locust olfactory system using in-vivo calcium imaging. We discover crowding-dependent synergistic interactions between food-related and social odors distributed across stable combinatorial response maps. The observed synergy was specific to the gregarious phase and manifested in distinct odor response motifs. Our results suggest a crowding-induced modulation of the locust olfactory system that enhances food detection in swarms. Overall, we demonstrate how linking sensory adaptations to behaviorally relevant tasks can improve our understanding of social modulation in non-model organisms.

Population Density-Dependent Developmental Regulation in Migratory Locust.

Insect development is intricately governed by hormonal signaling pathways, yet the pivotal upstream regulator that potentiates hormone activation remains largely elusive. The migratory locust, Locusta migratoria, exhibits population density-dependent phenotypic plasticity, encompassing traits such as flight capability, body coloration, and behavior. In this study, we elucidated a negative correlation between population density and ontogenetic development during the nymphal stage of locusts. We found that the level of density influences the developmental trajectory by modulating transcript abundance within the ecdysone signaling pathway, with knockdown of the prothoracicotropic hormone (PTTH) resulting in developmental delay. Transcriptomic analysis of locust brains across solitary and gregarious phases revealed significant differential expression of genes involved in various pathways, including protein synthesis, energy metabolism, hormonal regulation, and immunity. Notably, knockdown experiments targeting two energy regulators, adipokinetic hormone (AKH) and insulin-like polypeptide 1 (ilp1), failed to elicit changes in the developmental process in solitary locusts. However, knockdown of immunoglobulin (IG) significantly shortened the developmental time in higher-density populations. Collectively, our findings underscore the regulatory role of population density in determining developmental duration and suggest that an immune-related gene contributes to the observed differences in developmental patterns.

Olfactory Mating Signals in the Migratory Locust Locusta migratoria.

Swarming locusts cause huge plagues across the world threatening food production. Before swarms form, locust populations exhibit a dramatic phase change from a solitary to a gregarious phase. The cause of this phase change is a complicated interplay of conspecific and environmental cues and is, especially for one of the major pests, the migratory locust Locusta migratoria, still not well understood. Here we study the behavior of both solitary and gregarious L. migratoria towards the headspace odors of conspecifics. As we do not find a general attraction of gregarious animals to the headspace of gregarious conspecifics, swarm formation does not seem to be mainly governed by olfactory aggregation cues. When testing for potential mating signals, we observe that the headspace of virgin gregarious females is highly attractive only towards virgin males of the same phase, while mated gregarious males and solitary males, regardless of their mating state, do not become attracted. Interestingly, this phase-specific attraction goes along with the finding, that mating behavior in experiments with inter-phasic pairings is extremely rare. Our data suggest that odor emissions in L. migratoria play a significant role in a mating context.

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.

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.

Social recapitulation: Moulting can restore social tolerance in aggressive spiderlings.

In many taxa, the subsocial route is considered the main pathway to permanent sociality but the relative contribution of offspring interactions and parental care to the maintenance of cohesion and tolerance at advanced developmental stages remains poorly studied. Spiders are relevant models for this question because all exhibit a transient gregarious phase prior to dispersal and permanent sociality, which involves about 20 of the 50,000 species, is assumed to rely on the subsocial route. Using spiderlings of the solitary species Agelena labyrinthica, we manipulated the social context to demonstrate that tolerance in aggressive juveniles can be restored when exposed to siblings after moulting. We propose that moulting can reopen closed critical periods and renew the imprinting to social cues and thus lead to the reacquisition of tolerance. Our study highlights the critical role of contacts between juveniles in the expression of tolerance, which opens novel avenues for understanding social transitions.

Effect of entomopathogenic nematodes Steinernema species (steinernematidae: rhabditida) and Heterorhabditis bacteriophora (heterorhabditidae: rhabditida) on the digestive enzymes and midgut histology of the African migratory locust Locusta migratoria migratorioides (acrididae: orthoptera)

In some governorates of Egypt, the African migratory locust L. migratoria migratorioides is in a gregarious phase. Swarm development was successfully prevented by biological control agents. In this work, the two entomopathogenic nematode species, Steinernema sp. (SII)and Heterorhabditis bacteriophora (HP88) were investigated as natural enemies against the fifth nymph and adult African locusts at various concentrations (300, 600, 900, 1200, and 1500 Infected juveniles/100 g. soil). To ascertain fatal activity at 25 °C, the nematode-inoculated sand method was used. The two nematode species were semi-field administered against fifth nymphs and adult stages at (25 ± 2 °C) and 55–60% relative humidity with concentrations (3000, 6000, 9000, 12000, and 15000 Infected juveniles/kg. soil). Protease, amylase, invertase, and trehalase levels in the digestive enzymes of both fifth nymphs and adults fed with LC50 of both nematodes significantly decreased, but lipase and chitinase levels significantly increased. Adult locusts treated with the LC50 of S. sp. SII had basophilic epithelial cells, severe lumen hemorrhage, and highly aberrant proliferating cytoplasm, whereas the LC50 of H. bacteriophora HP88 displayed necrosis in an epithelial cell with vacuoles, loss of nucleus, and loss of goblet cells.

Analytical observation Upsurge and Current Situation of Desert Locust (Schistocerca gregaria) in Sindh, Pakistan:

Schistocerca gregaria (Forskl, 1775), one of the most notorious insects in the world, significantly harms the economy and agriculture each year. It was reputedly the biggest infestation to hit Pakistan since the 1990s, destroying wheat, rice, sugarcane, cotton, and vegetable crops, and it was also the cause of the worst disaster of 2019–2020. We have extensively examined the external characteristics of this swarm using the mitochondrial Cytochrome C Oxidase subunit 1 (COI), morphometry of the solitary and gregarious phases, influenced host plants, as well as its deterrent status. Swarms of Desert Locusts, which have been on the rise recently, have had a substantial negative influence on Pakistan's agriculture, destroying all kinds of crops. Preliminary projections of the financial losses over the two agricultural seasons in 2020 and 2021 may vary from 3.4 billion US dollars to 10.21 billion US dollars. This drastically increased the price of ordinary products in the market. In addition, locust activity has grown and is presently present in a number of affected areas. For the species to be managed and controlled effectively, accurate species identification is essential. This study seeks to explain this essential management attention

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.

The time course of behavioural phase change in the Central American locust Schistocerca piceifrons.

Locusts exhibit an extreme form of phenotypic plasticity and can exist as two alternative phenotypes, known as solitarious and gregarious phases. These phases, which can transform from one to another depending on local population density, show distinctly different behavioural characteristics. The proximate mechanisms of behavioural phase polyphenism have been well studied in the desert locust Schistocerca gregaria and the migratory locust Locusta migratoria, and what is known in these species is often treated as a general feature of locusts. However, this approach might be flawed, given that there are approximately 20 locust species that have independently evolved phase polyphenism. Using the Central American locust Schistocerca piceifrons as a study system, we characterised the time course of behavioural phase change using standard locust behavioural assays, using both a logistic regression-based model and analyses of separate behavioural variables. We found that for nymphs of S. piceifrons, solitarisation was a relatively fast, two-step process, but that gregarisation was a much slower process. Additionally, the density of the gregarisation treatment seemed to have no effect on the rate of phase change. These data are at odds with what we know about the time course of behavioural phase change in S. gregaria, suggesting that the mechanisms of locust phase polyphenism in these two species are different and may not be phylogenetically constrained. Our study represents the most in-depth study of behavioural gregarisation and solitarisation in locusts to date.

Ontogenetic change in social context as a cue for a behavioural switch in spiderlings

In many taxa, variations in the early social context are well known to influence the expression of a suite of behaviours later in life. For species living temporarily in groups, relatively few studies have examined how the ontogenetic change in the social context experienced by individuals during their transition from social to solitary influences their responses to environmental variation. It can be hypothesized that the presence of conspecifics during the gregarious phase may constrain the expression of any behavioural changes in response to external variations that would be detrimental to group cohesion and reduce the benefits associated with social life. In this study, we investigated the interplay between early social experience, satiation and ontogeny in the development of aggressive behaviours in juveniles of the solitary spider Agelena labyrinthica . First, we exposed spiderlings to a period of social isolation of similar duration at different ages, while others were continuously reared socially. Second, we examined how feeding influences the onset of agonistic interactions in spiderlings reared in isolation or socially. When spiders were socially deprived, we found that aggressive behaviour towards siblings increased both when they experienced isolation at a later developmental stage and when they were fed. This was in contrast to spiders maintained in groups, which, regardless of their age and whether they were fed or not, remained tolerant. Overall, our study suggests that changes in the social environment during ontogeny set the context for the expression of behavioural plasticity that prepares individuals for living alone. • Late social isolation during ontogeny increases aggression in spiderlings. • Food intake in socially isolated spiderlings increases aggression. • Spiderlings reared socially are less sensitive to environmental conditions. • The expression of behavioural plasticity depends on the social context.

The role of inter-individual intolerance in group cohesion and the transition to sociality in spiders.

Conspecific tolerance is key for maintaining group cohesion in animals. Understanding shifts from conspecific tolerance to intolerance is therefore important for understanding transitions to sociality. Subsocial species disperse to a solitary lifestyle after a gregarious juvenile phase and display conspecific intolerance as adults as a mechanism to maintain a solitary living. The development of intolerance towards group members is hypothesized to play a role in dispersal decisions in subsocial species. One hypothesis posits that dispersal is triggered by factors such as food competition with the subsequent development of conspecific intolerance, rather than conspecific intolerance developing prior to and potentially driving dispersal. Consistent with this hypothesis, we show that intolerance (inferred by inter-individual distance) developed post-dispersal in the subsocial spider Stegodyphus lineatus. The development of conspecific intolerance was delayed when maintaining spiders in groups showing plasticity in this trait, which is advantageous when trade-offs are not fixed over time. However, major evolutionary transitions, such as the transition to sociality, can permanently modify trade-offs and cause derived adaptations by the evolution of new or modified traits or evolutionary loss of traits that become redundant. Sociality in spiders has evolved repeatedly from subsocial ancestors, and social life in family groups combined with a lack of interaction with competing groups suggests relaxed selection for the development of conspecific intolerance. In the social Stegodyphus sarasinorum we found no evidence for the development of conspecific intolerance, consistent with the loss of this trait. Instead, we found evidence for conspecific attraction, which is likely to govern group cohesion.

Microbiome-related aspects of locust density-dependent phase transition.

Locust plagues are a notorious, ancient phenomenon. These swarming pests tend to aggregate and perform long migrations, decimating cultivated fields along their path. When population density is low, however, the locusts will express a cryptic, solitary, non-aggregating phenotype that is not considered a pest. Although the transition from the solitary to the gregarious phase has been well studied, associated shifts in the locust's microbiome have yet to be addressed. Here, using 16S rRNA amplicon sequencing, we compared the bacterial composition of solitary desert locusts before and after a phase transition. Our findings revealed that the microbiome is altered during the phase transition, and that a major aspect of this change is the acquisition of Weissella (Firmicutes). Our findings led us to hypothesize that the locust microbiome plays a role in inducing aggregation behaviour, contributing to the formation and maintenance of a swarm. Employing a mathematical model, we demonstrate the potential evolutionary advantage of inducing aggregation under different conditions; specifically, when the aggregation-inducing microbe exhibits a relatively high horizontal transmission rate. This is the first report of a previously unknown and important aspect of locust phase transition, demonstrating that the phase shift includes a shift in the gut and integument bacterial composition.

Can CRISPR help control locust populations to reduce the impact of plague outbreaks?

Desert locusts (Schistocerca gregaria) have been threatening food security since time immemorial and affecting human lives by destroying agriculture. Ravaging plague outbreaks are a reality and cause massive devastation across farmlands and pastures to this present day. Locust attacks and outbreaks globally affect vast areas and millions of people resulting in billions of dollars of economic loss. Although controlling the locust population through chemical pesticides is the primary method currently used, it unfortunately has not been highly effective in managing these outbreaks. Advancements in genetic engineering using CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and its successful experiments on arthropods provide a huge opportunity that can be extended to design similar genetic behavioral changes in locusts. CRISPR has become one of the most accurate, quick, and cost-effective techniques that could potentially be very effective in managing locust populations by editing specific genes, particularly those responsible for uncontrolled reproduction during their gregarious phase. Experiments conducted on Mosquitos (Aedes aegypti) to silence microRNA-309 (miR-309), permitting stage-specific degradation of its ability to reproduce in their blood-feeding triggered phase indicates that these interventions could also be performed successfully on desert locusts. It should be possible to achieve similar results in female locusts by targeting the miR-309~6 gene cluster, impairing their ovarian development and potentially controlling locust populations.

Allocation of more reproductive resource to egg size rather than clutch size of gregarious desert locust (Schistocerca gregaria) through increasing oogenesis period and oosorption rate

The desert locust, Schistocerca gregaria, shows a density-dependent reproductive trade-off by laying fewer but larger eggs in crowded conditions (gregarious phase) than in isolated conditions (solitarious phase). However, the physiological mechanisms controlling reproductive resource allocation remain unclear. We examined how egg production processes, including ovulation timing (i.e., oogenesis period), oocyte and ovarian growth rates, and oosorption rate (resorbing developing terminal oocytes), regulate reproductive outputs (egg biomass per clutch, egg size, and clutch size) during a reproductive cycle in S. gregaria by rearing them either under isolated or crowded conditions. We observed a common density-dependent negative correlation between egg size and clutch size, with no significant difference in egg biomass between the two rearing conditions. Dissection of female locusts after different days of oviposition revealed that the daily oocyte growth rate was almost similar between the two rearing conditions, but crowd-reared females ovulated later than isolated-reared ones, resulting in further oocyte growth in the former. Terminal oocytes were renewed by previous penultimate oocytes at the onset of a new reproductive cycle, and oosorption mainly occurred at an early stage in both rearing conditions; however, crowd-reared locusts displayed higher levels of oosorption compared to their isolated-reared counterparts. Crowding induced a high oosorption rate, resulting in a reduced clutch size and a prolonged oogenesis period, which in turn allowed oocytes to intake more yolk, which was probably recycled via oosorption. These results suggest that the length of the oogenesis period and oosorption rate are manipulated by maternal density, and these physiological modifications interactively regulate reproductive trade-off in S. gregaria.

Distribution, ecological and taxonomic study of Locusta migratoria migratoria (Linnaeus, 1758) and long term monitoring of its outbreak in Cold desert of Ladakh (UT) India

Short horned orthopetra from the family acrididae in gregarious phase is generally known as locust. In solitary condition they are harmless and overlooked. In history no epidemic of locust was snoop in the cold desert of Ladakh. In the last two decades Locusta migratoria migratoria (Linnaeus, 1758) is enormously affecting the livelihood of the trans-Himalayan region. Present study emphasized in Changthang, Chusul and Pangom Tso area of the Ladakh from 2008 to 2019. The purpose of this study was to explore the distribution, ecological and taxonomical aspects of the outbreak of migratory locusts in the fragile ecosystem. The long term monitoring assessed the geographic coverage of the locust infestation, swarming hotspots; evaluate the population parameters such as densities and developmental stages in the region. It was observed that migratory locust impinge along the Indus river area from Tibet and China and swarms year to year in Ladakh. They drastically impacted on the livelihood of the region with demolish the greenery of the area.

Larvae Crowding Increases Development Rate, Improves Disease Resistance, and Induces Expression of Antioxidant Enzymes and Heat Shock Proteins in Mythimna separata (Lepidoptera: Noetuidae).

High population density (crowding) becomes a stress factor in insects. The oriental armyworm, Mythimna separata (Walker), displays gregarious and solitary phases at high and low population densities, respectively. In this study, we compared life history, disease resistance, and induction of antioxidant enzymes and heat shock protein (HSPs) in two phases of M. separata larvae. Results showed that gregarious larvae had a faster growth rate and lower pupal weight compared to solitary larvae. Furthermore, gregarious individuals exhibited higher survival rates than solitary individuals after Beauveria bassiana infection. The gregarious larvae had higher malondialdehyde content compared to solitary ones, but no differences in total antioxidant capacity were observed between the two larval phases before or after infection. Superoxide dismutase and glutathione peroxidase activities were significantly lower in gregarious M. separata larvae than solitary individuals before infection, but no difference was detected in two phases after infection. However, peroxidase and catalase activities in the two phases showed no difference either before or after infection. Hsp19.8 and Hsp90 expression in gregarious larvae were up-regulated when compared to solitary individuals before or after infection. CuZnSOD expression was not different between the two phases before infection, but it was up-regulated in gregarious ones compared to solitary ones after infection. However, expression of other stress-related genes in gregarious larvae was either repressed or unchanged when compared to solitary individuals before or after infection. Thus, larval crowding changed life history, improved disease resistance of M. separata larvae, and induced variable response of antioxidant enzymes and HSPs to fungal infection.