Locusta Migratoria Research Articles

Occurrence of giant migratory locust Locusta migratoria (Acrididae) on Tsushima Island, Japan

This study focused on the populations of Locusta migratoria (Linnaeus, 1758) on Tsushima Island, Japan, which exhibited the largest mean adult body size when compared to other populations from various climatic regions. Certain individuals, designated as “giant locusts,” displayed exceptionally large sizes, with females and males identified when with head widths surpassing 10.5 mm and 8.0 mm, respectively. The maximum weight recorded for a giant locust was 8.9 g, in contrast to other examined females with the maximum weight ranging from 3.1 to 5.5 g. Notably, giant females exhibited the ability to yield larger egg pods and progeny compared to their counterparts. A positive correlation emerged between body size of adult females and the number of ovarioles. To explore the potential heritability of body size, selection experiments were conducted across two generations. A significant impact of selection on adult body size was apparent at LD 12:12h, whereas this effect was not evident at LD 16:8h. Furthermore, crossing experiments showed that body size at hatching closely resembled that of the female parent or demonstrated dimensions intermediary between the two parents, suggesting a complex genetic basis for the observed body size variations. This study provides no evidence of genetic differentiation between the giant locusts and the other locusts on this island.

Comparison of Inclusion of Skimmed Milk and Edible Insects on the Nutritional, Sensory and Microbial Qualities of Enriched High Energy Rice Biscuits

Aims: The growing world population, carbon foot prints of rearing conventional protein sources, issues of protein malnutrition (particularly in developing nations) and the urgent needs of protein-rich food relief materials in crisis situations may necessitate the utilisation of underutilised sustainable, nutritious protein sources. The study was conducted to investigate the possibility of using indigenous food crops and substituting milk powder with edible insects’ powder as a sustainable alternative protein source in high energy rice biscuits.
 Study Design: Biscuits were produced following the standard USAID recipe for HEB and comparisons were made with the insect biscuits and STD.
 Place and Duration of Study: Department of Food Science and Technology, Federal University of Technology, Akure, Ondo State, Nigeria, between January 2018 and August 2018.
 Methodology: The high energy biscuits was evaluated for proximate composition, total sugars using Anthrone reagent, micronutrients (vitamins and minerals), physical properties such as spread ratio, weight, width etc. Microbial and sensory quality of the biscuits were also determined using standard methods.
 Results: From the findings of the study, rice biscuits with silkworm pupae had outstanding protein (12.76 g/100 g), fat (15.78 g/100 g) and energy (451.38 Kcal/100 g) contents while biscuit with migratory locust powder had slightly higher total ash and CHO. The spread ratios of the biscuits were within acceptable range. In terms of mineral composition, all the insect biscuits were significantly superior to the milk biscuit except for calcium. However, all the rice HEB exceeded the minimum USAID specifications for nutritive elements. The insect biscuits had more than quadruple the recommended pro-vitamin A content (1200. 67 and 2390.05 μg/100 g) and impressive amounts of vitamin C (94.19 and 52.39 mg/100 g). The microbial assessment revealed HEB with remarkable quality signifying wholesome raw materials and hygienic sample preparation. All samples received good sensorial ranking, but HEB with migratory locust was preferred.
 Conclusion: The results demonstrated that edible insects could serve as alternative protein source in the production of a nutritious high energy biscuit with good organoleptic and microbiological qualities.

Nuclear Receptor FTZ-F1 Controls Locust Molt by Regulating the Molting Process of Locusta migratoria

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

Impact of In Vitro Gastrointestinal Digestion on the Phenolic Bioaccessibility and Bioactive Properties of Insect-Containing Beef Burgers.

Mealworm, migratory locust, and house cricket have recently been recognized by the European Commission as novel foods, thus being suitable in different food applications. In this work, we tested their powders as meat extenders at 5% (w/w) inclusion in beef burgers, considering their ability to vehicle phenolic compounds during simulated in vitro static gastrointestinal digestion (INFOGEST). Insect powders were abundant in different phenolic classes, recording the highest values in locust (LP; 314.69 mg/kg), followed by cricket (CP; 113.3 mg/kg) and mealworm (MWP; 51.9 mg/kg). Following a pan-cooking process, LP burgers were confirmed as the best source of phenolics, with a marked abundance of flavonoids and phenolic acids. Interestingly, the insect powders were found to affect the in vitro gastrointestinal bioaccessibility of phenolic compounds when compared with the CTR burger, likely promoted by the interactions between the phenolic compounds and proteins characterizing the tested insect powders. Among the most discriminant phenolic metabolites at the gastrointestinal level, we found several phenolic acids (mainly hydroxycinnamics), recording the highest content for the digested CP-containing burgers. Finally, stilbenes showed significant correlation values at the intestinal level with both antioxidant and enzymatic activities, while total flavonoids were the most correlated with the inhibition of acetylcholinesterase. Taken together, our preliminary findings demonstrated that insect powders added to beef burgers can promote the bioaccessibility and potential bioavailability of phenolics in the distal tracts of the intestine.

DNA Barcoding of Locusta migratoria manilensis (Orthoptera: Acrididae) Reveals Insights into the Species and Subspecies Differentiation

Abstract Accurate identification and classification of insect species, especially those with significant economic and ecological implications, have historically presented challenges. Migratory locusts, Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae), are notorious for their destructive impact on crops. Traditional morphological methods often face limitations in distinguishing closely related species and require taxonomic expertise. However, the emergence of DNA barcoding as a powerful tool for species identification has revolutionized the field of entomology. DNA barcoding utilizes a standardized DNA sequence, a molecular barcode, which serves as a distinct genetic signature for rapid and accurate species identification. In this study, DNA barcoding techniques were employed to identify and differentiate the migratory locust subspecies manilensis, in both its solitary and gregarious forms, as well as to determine its phylogenetic relationship with other related species within the Acrididae family. GenBank reference sequences were used to identify the locusts at the molecular subspecies level. Although the COI marker did not exhibit significant differences between the solitary and migratory forms, it was valuable in resolving the identification of L. migratoria subspecies. This lack of significant differences may be attributed to limited genetic variation of COI at the subspecies level and substantial genetic similarities between the solitary and migratory forms, likely stemming from a recent common ancestor. Nonetheless, using COI remains beneficial for subspecies identification in migratory locusts.

Osiris17 is indispensable for morphogenesis of intestinal tract in Locusta migratoria

The Osiris gene family is believed to play important roles in insect biology. Previous studies mainly focused on the roles of Osiris in Drorophila, how Osiris operates during the development of other species remains largely unknown. Here, we investigated the role of LmOsi17 in development of the hemimetabolous insect Locusta migratoria. LmOsi17 was highly expressed in the intestinal tract of nymphs. Knockdown of LmOsi17 by RNA interference (RNAi) in nymphs resulted in growth defects. The dsLmOsi17-injected nymphs did not increase in body weight or size and eventually died. Immunohistochemical analysis showed that LmOsi17 was localized to the epithelial cells of the foregut and the gastric caecum. Histological observation and hematoxylin-eosin staining indicate that the foregut and gastric caecum are deformed in dsLmOsi17 treated nymphs, suggesting that LmOsi17 is involved in morphogenesis of foregut and gastric caecum. In addition, we observed a significant reduction in the thickness of the new cuticle in dsLmOsi17-injected nymphs compared to control nymphs. Taken together, these results suggest that LmOsi17 contributes to morphogenesis of intestinal tract that affects growth and development of nymphs in locusts.

A neural m6A pathway regulates behavioral aggregation in migratory locusts.

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

Locustamigratoria (L.) (Orthoptera) in a warming world: unravelling the ecological consequences of climate change using GIS.

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.

RNAi-mediated silencing of the neverland gene inhibits molting in the migratory locust, Locusta migratoria

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.

Adipokinetic hormone signaling regulates adult dehydration resistance in the migratory locust

Drought events have become more severe under climate change, and this can pose a major threat to the survival of various organisms. The molecular mechanisms involved in dehydration resistance are not well known. Here, adults of the migratory locust, Locusta migratoria, were subjected to food-mediated dehydration, and adipokinetic hormone (AKH) signaling was found to play a key role in regulating dehydration resistance. Specifically, dehydration shortened the lifespan, increased the body weight loss, and reduced the water loss rate in adult locusts. Global transcriptome profiles revealed variations in tissue-specific gene expression between dehydration-resistant locusts and normal locusts. Importantly, dehydration selection and exposure induced prominent expression of AKH genes in the retrocerebral complex of adult locusts. Furthermore, individual knockdown of AKH1, AKH2, or AKH receptor (AKHR) accelerated water loss and shortened the lifespan of adult locusts under dehydration conditions, and trehalose supplementation ameliorated the negative effects caused by interference with AKH or AKHR. These findings demonstrated that AKH/AKHR signaling-dependent trehalose metabolism plays a crucial role in regulating locust dehydration resistance and thus provide novel insights into the regulatory mechanism underlying drought resistance.

Characterisation of the myosin light chain kinase (MLCK) gene of Locusta migratoria and the encoded MLCK.

Myosin light chain kinase (MLCK) is a dedicated kinase of myosin regulatory light chain (RLC), playing an essential role in the regulation of muscle contraction and cell motility. Much of the knowledge about MLCK comes from the study of vertebrate MLCK, and little is known about insect MLCK. Here, we identified the single MLCK gene in the locust Locusta migratoria, which spans over 1400 kb, includes 62 exons and accounts for at least five transcripts. We found that the five distinct transcripts of the locust MLCK gene are expressed in a tissue-specific manner, including three muscle-specific isoforms and two generic isoforms. To characterise the kinase activity of locust MLCK, we recombinantly expressed LmMLCK-G, the smallest locust MLCK isoform, in insect Sf9 cells. We demonstrated that LmMLCK-G is a Ca2+ /calmodulin-dependent kinase that specifically phosphorylates serine 50 of locust muscle myosin RLC (LmRLC). Additionally, we found that almost all LmRLC molecules in the flight muscle and the hindleg muscles of adult locusts are phosphorylated.

The mechanical properties of different cross-veins in the hind wing of locust Locusta migratoria under uniaxial tensile and stress relaxation tests.

Locust Locusta migratoria exhibits remarkable aerial performances, relying predominantly on its hind wings that generate most of lift and thrust for flight. The mechanical properties of the cross-veins determine the deformation of the hind wing, which greatly affect the aerodynamic performance of flapping flight. However, whether the mechanical behaviours of the locust cross-veins change with loading rate is still unknown. In this study, cross-veins in four physiological regions (anterior-medial, anterior-lateral, posterior-medial and posterior-lateral) of the hind wing from adult locusts were investigated using uniaxial tensile test, stress relaxation test and fluorescence microscopy. It was found that the cross-veins were a type of viscoelastic material (including rate-independent elastic modulus and obvious stress relaxation). The cross-veins in the two anterior regions of the hind wing had significantly higher elastic moduli and higher ultimate tensile stress than those of its two posterior regions. This difference might be attributed to different resilin distribution patterns in the cross-veins. These findings furnish new insights into the mechanical characteristics of the locust cross-veins, which might deepen our understanding of the aerodynamic mechanisms of locust flapping flight.

English

Background | With the global population on the rise, edible insects are considered a potential solution to food security, although concerns about risks such as anaphylaxis exist. Methods | 2,014 participants underwent testing with the Allergy Explorer-ALEX-2 including extracts of three novel foods: Acheta Domesticus (Ad), Locusta migratoria (Lm), and Tenebrio molitor (Tm). The IgE-mediated sensitization status was investigated in participants who had never knowingly consumed these insects. Data was recorded using an electronic database. Results | 195 individuals (9.7% of all participants) were sensitized to insects. Tropomyosin was co-recognized by 34%, and 18.5% were positive for arginine kinases. Reactivity to Sarcoplasmic-CB, Troponin-C, Paramyosin, or Myosin-light-chain was found in less than 5% of the population, whereas 108 individuals (55.4%) did not show any reactivity to invertebrate panallergens. Additionally, 33 individuals (16.9%) exhibited monosensitization exclusively to insects. Multivariate analysis revealed an inverse association between arachnid reactivity and sensitization to insect allergens, while Mollusca, Blattoidea, and tropomyosin reactivity displayed a direct relationship. Furthermore, Myosin-light-chain reactivity correlated with Ad and Lm, and Troponin-C with Ad and Tm sensitization. Conclusion | Edible insect extract IgE sensitization was observed in individuals without prior exposure to such foods. Mites showed a low likelihood of being primary sensitizers due to their inverse association with insect reactivity. Conversely, the direct association of insect sensitization with mollusk and cockroach extract reactivity suggests their potential as primary sensitizers in these participants. Tropomyosin consistently exhibited a positive association with reactivity to all studied insects, supporting its role as a primary sensitizer.

Neuropeptide ACP is required for fat body lipid metabolism homeostasis in locusts.

Fat body metabolism plays crucial roles in each aspect of insect life traits. Although neuropeptides have been documented to be one of the major neuroendocrinal regulators involved in fat body metabolism, the detailed regulatory mechanism is poorly explored. Here, we conducted comparative metabolome and transcriptome analyses of fat body between wide type (WT) and adipokinetic hormone/corazonin-related peptide (ACP) loss of function mutants of the migratory locust, Locusta migratoria. We found that knockout of ACP resulted in significantly reduced fat body triacylglycerol content but enhanced abundance of phospholipids, particularly phosphatidylcholine and phosphatidylethanolamine. Additionally, the expression levels of genes involved in triacylglycerol and phospholipid synthesis and degradation were significantly altered in the fat body of ACP mutants. Moreover, female ACP mutants displayed much higher fecundity compared to WT females. These findings highlight the important role of neuropeptide ACP in fat body lipid metabolism homeostasis in locusts.

The distinctive effect of different insect powders as meat extenders in beef burgers subjected to cooking and in vitro gastrointestinal digestion

Mealworm (MWP), migratory locust (LP), and house cricket (CP) are novel foods recently authorized by the European Commission. This work tested their powders as meat extenders at 5% inclusion in beef burgers. Insect powders were abundant in phenolics, recording the highest values in LP (1184.9 μg/g). The sensory analysis highlighted a higher visual and olfactory acceptability for MWP-burgers, followed by CP- and LP-burgers, whereas the texture of cooked burgers remained unaffected. Following pan-cooking, MWP-burgers and control exhibited comparable chemical profiles, while a significant down-accumulation of the heterocyclic amine 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline was observed in CP-burgers. In vitro gastrointestinal digestion highlighted metabolomic trends like control for MWP- and LP-burgers. In contrast, a reduced accumulation of lipids and increased content of dipeptides like glutaminylarginine (possibly acting as enzyme modulators) was observed for the CP-burgers.

Knickkopf (LmKnk) is required for chitin organization in the foregut of Locusta migratoria.

The foregut, located at the front of the digestive tract, serves a vital role in insects by storing and grinding food into small particles. The innermost layer of the foregut known as the chitinous intima, comes into direct contact with the food and acts as a protective barrier against abrasive particles. Knickkopf (Knk) is required for chitin organization in the chitinous exoskeleton, tracheae and wings. Despite its significance, little is known about the biological function of Knk in the foregut. In this study, we found that LmKnk was stably expressed in the foregut, and highly expressed before molting in Locusta migratoria. To ascertain the biological function of LmKnk in the foregut, we synthesized specific double-stranded LmKnk (dsLmKnk) and injected it into locusts. Our findings showed a significant decrease in the foregut size, along with reduced food intake and accumulation of residues in the foregut after dsLmKnk injection. Morphological observations revealed that newly formed intima became thinner and lacked chitin lamella. Furthermore, fluorescence immunohistochemistry revealed that LmKnk was located in the apical region of new intima and epithelial cells. Taken together, this study provides insights into the biological function of LmKnk in the foregut, and identifies the potential target gene for exploring biological pest management strategies.

Anatolian Short-Horned Grasshoppers Unveiled: Integrating Biogeography and Pest Potential.

Biogeographically, Anatolia harbours a rich diversity of short-horned grasshoppers (Orthoptera, Caelifera). The number of species recorded from Anatolia so far stands at 300. They inhabit diverse habitats ranging from arid Eremial to Euro-Siberian-like montane meadows, aligning with the topographical and climatological heterogeneity of Anatolia. Alongside some swarming species, the pest potential of several pullulating species needs attention. This is especially important concerning global warming, a scenario expected to be more severe in the Northern Mediterranean Basin in general and Anatolia specifically. A faunal list of biogeographic Anatolia, the area extending from the Aegean Sea in the west to the intermountain basin of the Caucasus in the northeast, the lowlands of Lake Urmia in the east, and Mesopotamia in the southeast, was developed. The recorded species were classified according to the phytogeographical provinces of Anatolia. Distributions of the species with the potential for pullulating were modelled using ecological-niche-modelling approaches for the present and future. The results have the potential to lead to the development of a concept that merges biogeography and the pest potential of certain Anatolian grasshopper species. Our results reveal the following: (i) Acrididae and Pamphagidae are the most diverse families represented in Anatolia; (ii) roughly 40% of Caelifera and 71% of Pamphagidae are endemics, suggesting Anatolia is a biodiversity hotspot; (iii) according to Caelifera diversity, the phytogeographical provinces of Anatolia follow an order of Irano-Anatolia, Euro-Siberia, Mediterranean, and Mesopotamia; and (iv) based on ecological modelling and personal observations, Dociostaurus maroccanus, Locusta migratoria, Calliptamus italicus, Heteracris pterosticha, Notostaurus anatolicus, Oedipoda miniata, and O. schochii should be monitored regarding their pest potential.

Comparison of the Virulence of Space Mutants of Aspergillus oryzae XJ-1 against Adult Locusta migratoria

Biological control methods provide a sustainable approach for reducing agricultural losses caused by locust plagues. Space mutagenesis can generate high numbers of mutations using satellites and spacecrafts, including beneficial and stable mutants. Aspergillus oryzae XJ-1 was recently reported to show high virulence against locusts. We subjected this fungal pathogen to space mutagenesis to obtain more effective strains. Pathogen conidia powder was mutated in the China Space Station for 6 months. We obtained five mutants of A. oryzae XJ-1, TQ201, TQ238, TQ302, TQ549, and TQ555, and all mutants were identified as A. oryzae by molecular techniques. TQ549 showed the highest virulence against adult L. migratoria (LT50: 4.97 ± 0.21 days); the LT50 of A. oryzae XJ-1 was 5.67 ± 0.06 days. Both TQ549 and A. oryzae XJ-1 grew most rapidly at 33 °C on potato dextrose agar (PDA) plates. There was no significant difference in the growth rate of TQ549 and A. oryzae XJ-1 at 24 °C. The colony morphological characteristics of the five mutants on PDA plates differed from that of A. oryzae XJ-1. The space mutant TQ549, which showed high virulence against adult locusts, could be used as a biological control agent for the control of locust infestations.

Exploiting Locusta migratoria as a source of bioactive peptides with anti-fibrosis properties using an in silico approach.

Edible insects have been proposed as an environmentally and economically sustainable source of protein, and are considered as an alternative food, especially to meat. The migratory locust, Locusta migratoria, is an edible species authorised by the European Union as a novel food. In addition to their nutritional value, edible insects are also sources of bioactive compounds. This study used an in silico approach to simulate the gastrointestinal digestion of selected L. migratoria proteins and posteriorly identify peptides capable of selectively inhibiting the N-subunit of the somatic angiotensin-I converting enzyme (sACE). The application of the molecular docking protocol enabled the identification of three peptides, namely TCDSL, IDCSR and EAEEGQF, which were predicted to act as potential selective inhibitors of the sACE N-domain and, therefore, possess bioactivity against cardiac and pulmonary fibrosis.

A Core Structural Protein That Builds the Locust Mandible with a Mechanical Gradient.

Natural materials, such as locust mandibles and squid beaks, define significant mechanical gradients that have been attributed to the chemical gradients of their specialized structural proteins (SPs). However, the mechanism by which SPs form chemical gradients in these materials remains unknown. In this study, a highly abundant histidine-rich structural protein (LmMHSP) was identified in the mandible of a migratory locust (Locusta migratoria). LmMHSP was proven by both in vivo and in vitro evidence to act as a core building block of the mandible with a variety of synergistic functions including chitin binding, matrix formation via liquid-liquid phase separation, chemical cross-linking, and metal coordination. Furthermore, we found that the SP gradient in the locust mandible stems from the chitin-binding activity of LmMHSP and different microstructures of chitin scaffolds in different regions. These findings advance our understanding of the formation mechanisms of natural biomaterials and have implications for the fabrication of biomimetic materials.