摘要: Ciliary neurotrophic factor (CNTF) acts as a potent neuroprotective agent in neuronal survival and regeneration, and can also induce the differentiation of several stem cells into neurons, which highlights the broad application of CNTF in biomedicine. However, large-scale production of bioactive recombinant human CNTF protein remains to be explored. Herein, this study aims to express a bioactive human CNTF protein on a large scale by genetically engineering a silk gland bioreactor of silkworm. Our results showed that CNTF protein was successfully expressed in the middle silk gland (MSG) of silkworm, which can be secreted into the silks with the amount of 3.2 mg/g cocoons. The fabrication of human CNTF-functionalized silk material was able to promote proliferation and migration of neural cells when compared to the natural silk protein. Importantly, this functional silk material could also facilitate neurite outgrowth of mouse retinal ganglion cell (RGC-5) cells. All these data demonstrated a high bioactivity of the recombinant human CNTF protein expressed in the MSG of silkworm. The further fabrication of different silk materials with CNTF bioactivity will give biomedical applications in tissue engineering and neuroregeneration. We have established an efficient strategy for the large-scale and cost-effective production of bioactive human ciliary neurotrophic factor (CNTF) in silkworm. The fabrication of human CNTF silk material can promote proliferation, migration, and neurite outgrowth of neural cells. These data will provide biomedical applications in tissue engineering and neuroregeneration by using functionalized CNTF materials. image

摘要: Agricultural pests can develop behavioral resistance to insecticides by choosing to feed or oviposit on insecticide-free hosts. As young larvae have relatively low mobility, oviposition preferences from female adults may play a critical role in shaping the evolutionary trajectory of pest populations. While oviposition avoidance of insecticide-treated hosts was found in different agriculture pests, it remains unclear whether female adults actively choose to occupy insecticide-free hosts. To address this question, we investigated feeding and oviposition preferences between imidacloprid-treated and imidacloprid-free plants in the Colorado potato beetle, Leptinotarsa decemlineata Say, a major potato pest. We performed behavioral choice assays on two strains that differed in both fecundity and insecticide resistance. We found that one strain preferred to feed on the insecticide-free plants and that this preference is not innate. Meanwhile, the other strain chose plants for feeding and oviposition randomly. Further analyses of the moving patterns of the beetles suggested that the oviposition preference in the first strain is likely due to active learning. A strain of L. decemlineata avoided imidacloprid-treated plants for feeding and oviposition in choice assays while another strain chose plants randomly. First feeding is random in both strains. The oviposition avoidance behavior might be an active response due to learning from feeding on both choices of plants. image

摘要: Insect legs play a crucial role in various modes of locomotion, including walking, jumping, swimming, and other forms of movement. The flexibility of their leg joints is critical in enabling various modes of locomotion. The frog-legged leaf beetle Sagra femorata possesses remarkably enlarged hind legs, which are considered to be a critical adaptation that enables the species to withstand external pressures. When confronted with external threats, S. femorata initiates a stress response by rapidly rotating its hind legs backward and upward to a specific angle, thereby potentially intimidating potential assailants. Based on video analysis, we identified 4 distinct phases of the hind leg rotation process in S. femorata, which were determined by the range of rotation angles (0(degrees)-168.77(degrees)). Utilizing micro-computed tomography (micro-CT) technology, we performed a 3-dimensional (3D) reconstruction and conducted relative positioning and volumetric analysis of the metacoxa and metatrochanter of S. femorata. Our analysis revealed that the metacoxa-trochanter joint is a screw-nut structure connected by 4 muscles, which regulate the rotation of the legs. Further testing using a 3D-printed model of the metacoxa-trochanter joint demonstrated its possession of a self-locking mechanism capable of securing the legs in specific positions to prevent excessive rotation and dislocation. It can be envisioned that this self-locking mechanism holds potential for application in bio-inspired robotics.

摘要: We attempt to determine the effect of the dietary switch from a native to non-native prey on the gut microbiota in the predaceous ladybird Harmonia axyridis larvae and adults and examine how the dietary effect may vary across generations. We fed H. axyridis with different diets, native aphid Megoura japonica (Matsumura) versus non-native mealybug Phenacoccus solenopsis (Tinsley), for 5 generations and sequenced microbes in the gut of the 3rd instar larvae and adults of the 1st, 3rd, and 5th generations. In addition, we identified microbes in M. japonica and P. solenopsis. The 2 prey species differed in microbial community as measured by abundances of prevalent microbial genera and diversity. In H. axyridis, abundances of some prevalent microbial genera differed between the 2 diets in the 1st and 3rd generations, but the difference disappeared in the 5th generation; this tendency is more obvious in adults than in larvae. Overall, gut microbial assemblages became gradually cohesive over generations. Microbial diversity differed between diets in the 1st and 3rd generations but became similar in the 5th generation. Major prevalent gut microbial genera are predicted to be associated with metabolic functions of H. axyridis and are more abundant for consuming the mealybug than the aphid. Our findings from this study suggest that the gut microbiota in H. axyridis is flexible in response to the dietary switch, but tends toward homogeneity in microbial composition over generations.

摘要: Insects are the most diverse group of organisms in the animal kingdom, and some species exhibit complex social behaviors. Although research on insect object use is still in its early stages, insects have already been shown to display rich object-use behaviors. This review focuses on patterns and behavioral flexibility in insect object-use behavior, and the role of cultural evolution in the development of object-use behaviors. Object use in insects is not widespread but has been documented in a diverse set of taxa. Some insects can use objects flexibly and display various object-use patterns. Like mammals and birds, insects use objects in diverse activities, including foraging, predator defense, courtship, and play. Intelligence, pre-existing manipulative behaviors, and anatomical structure affect innovations in object use. In addition, learning and imitation are the main mechanisms underlying the spread of object-use behaviors within populations. Given that insects are one of the major animal groups engaging in object use, studies of insect object use could provide general insights into object use in the animal kingdom.

摘要: During the pupal-adult eclosion process of holometabolous insects, the old cuticle is shed and replaced by a completely different new cuticle that requires tanning and expansion, along with extensive extracellular matrix (ECM) remodeling. In vertebrates, matrix metalloproteinases (MMPs), a class of zinc-dependent endopeptidases, play key roles in regulating the ECM that surrounds cells. However, little is known about these extracellular proteinases available in insects. The small hive beetle (SHB), Aethina tumida, is a widespread invasive parasite of honey bees. In this study, 6 MMP homologs were identified in the SHB genome. RNA interference experiments showed that all 6 AtMmps are not required for the larval-pupal transition, only AtMmp2 was essential for pupal-adult eclosion in SHB. Knockdown of AtMmp2 resulted in eclosion defects and wing expansion failure, as well as mortality within 3 d of adult eclosion. Transcriptomic analysis revealed that knockdown of AtMmp2 significantly increased expression of the Toll and Imd pathways, chitin metabolism, and cross-linking (such as the pro-phenoloxidase activating cascade pathway and the tyrosine-mediated cuticle sclerotization and pigmentation pathway). These data revealed evolutionarily conserved functions of Mmp2 in controlling adult eclosion and wing expansion, also provided a preliminary exploration of the novel function of regulating Toll and Imd pathways, as well as new insights into how MMPs regulate insect development and defense barriers.

摘要: Insects show highly complicated adaptive and sophisticated behaviors, including spatial orientation skills, learning ability, and social interaction. These behaviors are controlled by the insect brain, the central part of the nervous system. The tiny insect brain consists of millions of highly differentiated and interconnected cells forming a complex network. Decades of research has gone into an understanding of which parts of the insect brain possess particular behaviors, but exactly how they modulate these functional consequences needs to be clarified. Detailed description of the brain and behavior is required to decipher the complexity of cell types, as well as their connectivity and function. Single-cell RNA-sequencing (scRNA-seq) has emerged recently as a breakthrough technology to understand the transcriptome at cellular resolution. With scRNA-seq, it is possible to uncover the cellular heterogeneity of brain cells and elucidate their specific functions and state. In this review, we first review the basic structure of insect brains and the links to insect behaviors mainly focusing on learning and memory. Then the scRNA applications on insect brains are introduced by representative studies. Single-cell RNA-seq has allowed researchers to classify cell subpopulations within different insect brain regions, pinpoint single-cell developmental trajectories, and identify gene regulatory networks. These developments empower the advances in neuroscience and shed light on the intricate problems in understanding insect brain functions and behaviors. Single-cell RNA sequencing (scRNA-seq) has emerged in the past decade and profoundly accelerated our understanding of brain complexity. scRNA-seq has a higher resolution than bulk RNA-seq, which enables novel cell type classification, dynamic trajectory construction, and gene regulatory network identification. To give a general understanding of scRNA-seq application in brain science, we depict a brief overview of the experimental workflow and a typical downstream data analysis in this graphical abstract.image

摘要: Drosophila suzukii is a notorious pest which causes devastating damage to thin-skinned fruits, and the larvae feed on the fruit, causing extensive agricultural economic loss. The current application of insecticides to manage this pest results in serious resistance and environmental hazards, so an alternative strategy for D. suzukii biocontrol is urgently needed. Here, we reported that entomopathogenic Bacillus cereus has the potential to biocontrol D. suzukii. We isolated and identified the bacterial strain, B. cereus H1, that was detrimental to the fitness of both D. suzukii progenies and parents. D. suzukii was robustly repelled to depositing eggs on the halves with metabolites of B. cereus H1. Both males and females of D. suzukii were susceptible to B. cereus H1. B. cereus H1 significantly arrested larval development with at least 40% lethal larvae. The median lethal time (LT50) of males and females of D. suzukii challenged with B. cereus H1 was 3 and 2 d, respectively. Moreover, B. cereus H1 disrupted the intestinal integrity and pH value of D. suzukii and resulted in an increase in bacterial load of guts and hemolymph. Mechanistically, infection of B. cereus H1 led to the activation of the dual oxidase (DUOX)-ROS-Jun N-terminal kinase (JNK) pathway. The findings showed that the entomopathogen B. cereus H1 could potentially act as a biological control agent against D. suzukii, advancing fundamental concepts of integrated pest management programs against D. suzukii.

摘要: Knowing how environmental conditions affect performance traits in pest insects is important to improve pest management strategies. It can be informative for monitoring, but also for control programs where insects are mass-reared, and field-released. Here, we investigated how adult thermal acclimation in sterile Bactrocera dorsalis affects dispersal and recapture rates in the field using a mark-release-recapture method. We also considered how current abiotic factors may affect recapture rates and interact with thermal history. We found that acclimation at 20 or 30 degrees C for 4 d prior to release reduced the number of recaptures in comparison with the 25 degrees C control group, but with no differences between groups in the willingness to disperse upon release. However, the deleterious effects of acclimation were only detectable in the first week following release, whereafter only the recent abiotic conditions explained recapture rates. In addition, we found that recent field conditions contributed more than thermal history to explain patterns of recaptures. The two most important variables affecting the number of recaptures were the maximum temperature and the average relative humidity experienced in the 24 h preceding trapping. Our results add to the handful of studies that have considered the effect of thermal acclimation on insect field performance, but notably lend support to the deleterious acclimation hypothesis among the various hypotheses that have been proposed. Finally, this study shows that there are specific abiotic conditions (cold/hot and dry) in which recaptures will be reduced, which may therefore bias estimates of wild population size.

摘要: The tetraspanin gene family encodes cell-surface proteins that span the membrane 4 times and play critical roles in a wide range of biological processes across numerous organisms. Recent findings highlight the involvement of a tetraspanin of the lepidopteran pest Helicoverpa armigera in resistance to Bacillus thuringiensis Cry insecticidal proteins, which are extensively used in transgenic crops. Thus, a better understanding of lepidopteran tetraspanins is urgently needed. In the current study, genome scanning in 10 lepidopteran species identified a total of 283 sequences encoding potential tetraspanins. Based on conserved cysteine patterns in the large extracellular loop and their phylogenetic relationships, these tetraspanins were classified into 8 subfamilies (TspA to TspH). Six ancestral introns were identified within lepidopteran tetraspanin genes. Tetraspanins in TspA, TspB, TspC, and TspD subfamilies exhibit highly similar gene organization, while tetraspanins in the remaining 4 subfamilies exhibited variation in intron loss and/or gain during evolution. Analysis of chromosomal distribution revealed a lepidopteran-specific cluster of 10 to 11 tetraspanins, likely formed by tandem duplication events. Selective pressure analysis indicated negative selection across all orthologous groups, with omega values ranging between 0.004 and 0.362. However, positive selection was identified at 18 sites within TspB5, TspC5, TspE3, and TspF10. Furthermore, spatiotemporal expression analysis of H. armigera tetraspanins demonstrated variable expression levels across different developmental stages and tissues, suggesting diverse functions of tetraspanin members in this globally important insect pest. Our findings establish a solid foundation for subsequent functional investigations of tetraspanins in lepidopteran species.