Silkworm Bombyx Mori Research Articles

Enhanced sensitivity of chimeric insect olfactory co-receptors for detecting odorant molecules

Insect olfactory receptors (ORs) are seven-transmembrane domain ion channels that function by forming heteromeric complexes with olfactory receptor co-receptors (Orcos). In this study, we investigated the potential for enhancing sensitivity of odor detection and responsivity through genetic modification of Orcos, considering its wider application in odor sensing. First, we measured the intensity of response to 1-octen-3-ol for the mosquito Aedes aegypti OR (AaOR8) when complexed individually with an Orco from the same mosquito (AaOrco), the honeybee Apis mellifera (AmOrco), the silkworm Bombyx mori (BmOrco), or the fruit fly Drosophila melanogaster (DmOrco). Relative to the other Orcos, AmOrco demonstrated higher sensitivity and responsivity, with a 1.8 to 21-fold decrease in the half-maximal effective concentration (EC50) and a 1.6–8.8-fold increase in the maximal effect (Emax), respectively. Furthermore, AmOrco co-expressed with AaOR10, BmOR56, or DmOR47a showed higher sensitivity and responsivity than AaOrco, BmOrco, or DmOrco co-expressed with their respective ORs. To further increase sensitivity and responsivity, we engineered chimeric Orcos by fusing AmOrco with DmOrco, considering the domain characteristics of Orcos. The response to 1-octen-3-ol was evaluated for AaOR8 when complexed individually with AmOrco, as well as for a mutant that combines DmOrco from the N-terminal (NT) to the C-terminal region of the fourth transmembrane domain (TM4) with the region of AmOrco following TM4 (Dm[NT-TM4]AmOrco). When compared to AmOrco, Dm(NT-TM4)AmOrco showed higher sensitivity and responsivity, with a 1.4-fold decrease in the EC50 and a 1.4-fold increase in the Emax, respectively. In addition, Dm(NT-TM4)AmOrco co-expressed with either DmOR47a or BmOR56 demonstrated higher sensitivity and responsivity than AmOrco co-expressed with their respective ORs. These results suggest that AmOrco could be a relatively more sensitive Orco, and further enhancement of sensitivity and responsivity could be achieved through recombination with heterologous Orcos near the TM4 of AmOrco.

Effect of acute exposure of Hg on physiological parameters and transcriptome expression in silkworms (Bombyx mori).

Mercury (Hg) contamination poses a global threat to the environment, given its elevated ecotoxicity. Herein, we employed the lepidopteran model insect, silkworm (Bombyx mori), to systematically investigate the toxic effects of Hg-stress across its growth and development, histomorphology, antioxidant enzyme activities, and transcriptome responses. High doses of Hg exposure induced evident poisoning symptoms, markedly impeding the growth of silkworm larvae and escalating mortality in a dose-dependent manner. Under Hg exposure, the histomorphology of both the midgut and fat body exhibited impairments. Carboxylesterase (CarE) activity was increased in both midgut and fat body tissues responding to Hg treatment. Conversely, glutathione S-transferase (GST) levels increased in the fat body but decreased in the midgut. The transcriptomic analysis revealed that the response induced by Hg stress involved multiple metabolism processes. Significantly differently expressed genes (DEGs) exhibited strong associations with oxidative phosphorylation, nutrient metabolisms, insect hormone biosynthesis, lysosome, ribosome biogenesis in eukaryotes, and ribosome pathways in the midgut or the fat body. The findings implied that exposure to Hg might induce the oxidative stress response, attempting to compensate for impaired metabolism. Concurrently, disruptions in nutrient metabolism and insect hormone activity might hinder growth and development, leading to immune dysfunction in silkworms. These insights significantly advance our theoretical understanding of the potential mechanisms underlying Hg toxicity in invertebrate organisms.

Comparative analysis of testicular fusion in Spodoptera litura (cutworm) and Bombyx mori (silkworm): Histological and transcriptomic insights

Spodoptera litura commonly known as the cutworm, is among the most destructive lepidopteran pests affecting over 120 plants species. The powerful destructive nature of this lepidopteran is attributable to its high reproductive capacity. The testicular fusion that occurs during metamorphosis from larvae to pupa in S.litura positively influences the reproductive success of the offspring. In contrast, Bombyx mori, the silkworm, retains separate testes throughout its life and does not undergo this fusion process. Microscopic examination reveals that during testicular fusion in S.litura, the peritoneal sheath becomes thinner and more translucent, whereas in B.mori, the analogous region thickens. The outer basement membrane in S.litura exhibits fractures, discontinuity, and uneven thickness accompanied by a significant presence of cellular secretions, large cell size, increased vesicles, liquid droplets, and a proliferation of rough endoplasmic reticulum and mitochondria. In contrast, the testicular peritoneal sheath of B.mori at comparable developmental stage exhibits minimal change. Comparative transcriptomic analysis of the testicular peritoneal sheath reveals a substantial difference in gene expression between the two species. The disparity in differential expressed genes (DEGs) is linked to an enrichment of numerous transcription factors, intracellular signaling pathways involving Ca2+ and GTPase, as well as intracellular protein transport and signaling pathways. Meanwhile, structural proteins including actin, chitin-binding proteins, membrane protein fractions, cell adhesion, extracellular matrix proteins are predominantly identified. Moreover, the study highlights the enrichment of endopeptidases, serine proteases, proteolytic enzymes and matrix metalloproteins, which may play a role in the degradation of the outer membrane. Five transcription factors-Slforkhead, Slproline, Slcyclic, Slsilk, and SlD-ETS were identified, and their expression pattern were confirmed by qRT-PCR. they are candidates for participating in the regulation of testicular fusion. Our findings underscore significant morphological and trancriptomic variation in the testicular peritoneal sheath of S.litura compared to the silkworm, with substantial changes at the transcriptomic level coinciding with testicular fusion. The research provides valuable clues for understanding the complex mechanisms underlying this unique phenomenon in insects.

Fatty Acid Profile and Thermal Behavior of Fat-Rich Edible Insect Oils Compared to Commonly Consumed Animal and Plant Oils.

This study compared the physicochemical properties of edible insect oils from silkworm (Bombyx mori) pupa (SP), sago palm weevil (Rhynchophorus ferrugineus) larva (PW), and bamboo caterpillar (Omphisa fuscidentalis; BC) to oils from chicken skin (CK), beef back fat (BF), pork back fat (PF), salmon belly (SB), sea bass belly (BB), coconut (C), and peanut (P). The fatty acid profiles and thermal behaviors (crystallization and melting) of the extracted oils were evaluated. PW and BC oils had more saturated fatty acids (SFAs) than CK, PF, SB, BB, and P oils. SP oil had equivalent SFA content to CK and BB oils. Insect oils exhibited similar monounsaturated fatty acid concentrations in all samples, except C oils. PW and BC oils exhibited a higher content of palmitoleic acid than the other oils. SP oils contained polyunsaturated fatty acids similar to those in SB and BB oils, which were higher than those in PW, BC, CK, BF, and PF oils. SP oil also exhibited the highest concentration of α-linolenic acid (C18:3 n-3). Arachidonic acid (0.01-0.02 g/100 g) in all insect oils was lower level compared to CK, BF, PF, SB, and BB oils. SP oil (0.03 g/100 g) exhibited a slightly higher level of eicosapentaenoic acid compared to PW (0.01 g/100 g) and BC (0.01 g/100 g) oils. The insect oils were liquid at ambient temperature, solid below -15°C, and required less energy (△Hm-max) for melting than other samples. This study indicated that insects, particularly SP, could serve as an alternative source of fat to meet its growing demand.

Let-7 microRNA targets BmCentrin to modulate the development and functionality of the middle silk gland in the silkworm, Bombyx mori.

The silk gland of the silkworm Bombyx mori serves as a valuable model for investigating the morphological structure and physiological functions of organs. Previous studies have demonstrated the notable regulatory role of let-7 microRNA in the silk gland, but its specific molecular mechanism remains to be elucidated across different segments of this organ. In this study, we further investigated the functional mechanism of let-7 in the middle silk gland (MSG). The MSG of a let-7 knockout strain was analyzed using a combined proteomic and metabolomic technique, revealing the enrichment of differential proteins and metabolites in the DNA synthesis and energy metabolism pathways. BmCentrin was identified as a novel target gene of let-7 in the MSG, and its downregulation inhibited the proliferation of BmN4-SID1 cells, which is exactly opposite to the role of let-7 in these cells. CRISPR/Cas9 genome editing and transgenic technologies were employed to manipulate BmCentrin in the MSG. Knockout of BmCentrin led to severe MSG atrophy, whereas the overexpression of BmCentrin resulted in beaded MSG. Further measurements of these knockout or overexpression strains revealed significant changes in the expression levels of sericin protein genes, the weight of the cocoon and the mechanical properties of the silk. Investigating the biological role of BmCentrin in the silk gland offers valuable insights for elucidating the molecular mechanisms by which let-7 controls silk gland development and silk protein synthesis in the silkworm.

MIG1, TUP1 and NRG1 mediated yeast to hyphal morphogenesis inhibition in Candida albicans by ganciclovir.

Candida albicans is a polymorphic human fungal pathogen and the prime etiological agent responsible for candidiasis. The main two aspects of C. albicans virulence that have been suggested are yeast-to-hyphal (Y-H) morphological transitions and biofilm development. Anti-fungal agents targeting these virulence attributes enhances the antifungal drug development process. Repositioning with other non-fungal drugs offered a one of the new strategies and a potential alternative option to counter the urgent need for antifungal drug development. In the current study, an antiviral drug ganciclovir was screened as an antifungal agent against ATCC 90028, 10231 and clinical isolate (C1). Ganciclovir at 0.5mg/ml concentration reduced 50% hyphal development on a silicon-based urinary catheter and was visualized using scanning electron microscopy. Ganciclovir reduced ergosterol biosynthesis in both strains and C1 isolate of C. albicans in a concentration-dependent manner. Additionally, a gene expression profile study showed that ganciclovir treatment resulted in upregulation of hyphal-specific repressors MIG1, TUP1, and NRG1 in C. albicans. Additionally, an in vivo study on the Bombyx mori silkworm model further evidenced the virulence inhibitory ability of ganciclovir (0.5mg/ml) against C. albicans. This is the first report that explore the novel anti-morphogenic activities of ganciclovir against the pathogenic C. albicans strains, along with clinical isolates. Further, ganciclovir may be considered for therapeutic purpose after combinations with standard antifungal agents.

The mechanoreceptor Piezo is required for spermatogenesis in Bombyx mori

BackgroundThe animal sperm shows high diversity in morphology, components, and motility. In the lepidopteran model insect, the silkworm Bombyx mori, two types of sperm, including nucleate fertile eupyrene sperm and anucleate unfertile apyrene sperm, are generated. Apyrene sperm assists fertilization by facilitating the migration of eupyrene spermatozoa from the bursa copulatrix to the spermatheca. During spermatogenesis, eupyrene sperm bundles extrude the cytoplasm by peristaltic squeezing, while the nuclei of the apyrene sperm bundles are discarded with the same process, forming matured sperm.ResultsIn this study, we describe that a mechanoreceptor BmPiezo, the sole Piezo ortholog in B. mori, plays key roles in larval feeding behavior and, more importantly, is essential for eupyrene spermatogenesis and male fertility. CRISPR/Cas9-mediated loss of BmPiezo function decreases larval appetite and subsequent body size and weight. Immunofluorescence analyses reveal that BmPiezo is intensely localized in the inflatable point of eupyrene sperm bundle induced by peristaltic squeezing. BmPiezo is also enriched in the middle region of apyrene sperm bundle before peristaltic squeezing. Cytological analyses of dimorphic sperm reveal developmental arrest of eupyrene sperm bundles in BmPiezo mutants, while the apyrene spermatogenesis is not affected. RNA-seq analysis and q-RT-PCR analyses demonstrate that eupyrene spermatogenic arrest is associated with the dysregulation of the actin cytoskeleton. Moreover, we show that the deformed eupyrene sperm bundles fail to migrate from the testes, resulting in male infertility due to the absence of eupyrene sperm in the bursa copulatrix and spermatheca.ConclusionsIn conclusion, our studies thus uncover a new role for Piezo in regulating spermatogenesis and male fertility in insects.

Biosynthesis of Pteridines in Insects: A Review

Pteridines are important cofactors for many biological functions of all living organisms, and they were first discovered as pigments of insects, mainly in butterfly wings and the eye and body colors of insects. Most of the information on their structures and biosynthesis has been obtained from studies with the model insects Drosophila melanogaster and the silkworm Bombyx mori. This review discusses, and integrates into one metabolic pathway, the different branches which lead to the synthesis of the red pigments “drosopterins”, the yellow pigments sepiapterin and sepialumazine, the orange pigment erythropterin and its related yellow metabolites (xanthopterin and 7-methyl-xanthopterin), the colorless compounds with violet fluorescence (isoxanthopterin and isoxantholumazine), and the branch leading to tetrahydrobiopterin, the essential cofactor for the synthesis of aromatic amino acids and biogenic amines.

Cocoon Care : Transformative Monitoring For Sericulture

Sericulture is the process of cultivating silkworms and extracting silk from them. The caterpillars of the domestic silk moth (also called „Bombyx Mori‟) are the most commonly used silkworm species in sericulture. Other types of silkworms (such as Eri, Muga, and Tasar) are also cultivated for the production of „wild silks‟. Silk is known as the queen of textiles due to its softness, durability, and luster. Furthermore, the silk fibers provide characteristics that are superior toany other type of fiber (e.g., water absorbency, heat resistance, dyeing efficiency, and luster). This textile is obtained from cocoons spun by larvae known as silkworm (Bombyx mori), which were discovered in China between 2600 and 2700 BC. The process of silk production is known as sericulture, beginning with the rearing of the silkworm1

Improvement of Silkworm (Bombyx mori L.) Growth and Cocoon Quality Using Nickel Sulfate Supplementation

The silkworm belongs to the order Lepidoptera and the family Bombycidae (Bombyx mori L.). The silkworm industry plays a crucial role in providing employment opportunities in India. This study investigated the effects of nickel sulfate supplementation on the growth and cocoon quality of silkworm (Bombyx mori L.). The experiment was conducted at a silkworm-rearing center in Palayamkottai, India, from December 2022 to January 2023. Silkworms were fed Victory-1 (V1) mulberry leaves soaked in different nickel sulfate solutions (100 ppm, 300 ppm, and 500 ppm). Larval weight, duration, cocoon weight, shell weight, shell ratio, and pupal weight were measured for 10 healthy silkworms with good quality cocoons selected from each treatment group. Compared to the control group, supplementation with 500-ppm nickel sulfate solution significantly improved silkworm growth performance (weight) and economic traits (cocoon weight and quality). GRAPHICAL ABSTRACT

Impact of Surface Functionality on Biodistribution of Gold Nanoparticles in Silkworms

To date, animal models are still indispensable for studying biodistribution and elimination of nanomaterials. However, the use of mammals for in vivo experiments faces various challenges including increasing regulatory hurdles and costs. This study aims to validate larvae of the domestic silkworm Bombyx mori as an alternative invertebrate model for preliminary in vivo research. Organ distribution and elimination of gold nanoparticles (AuNPs) are compared with four different surface functionalities in silkworms after systemic administration: AuNPs coated with poly(ethylene glycol) (PEG), with polyglycidols (PGs) that are slightly hydrophobic (PG(alkyl)), positively charged (PG(+)), or negatively charged (PG(−)). Subsequent inductive coupled plasma mass spectrometry 6 or 24 h after AuNPs administration reveals the biodistribution in silkworm hemolymph, midgut, epidermis, and excrements. Even after 24 h incubation, hemolymph contains the highest AuNPs concentrations, independent of surface functionalization indicating a prolonged circulation time and slow distribution into different silkworm organs and tissues. Positively charged PG(+)AuNPs show three times higher concentrations in the midgut and are excreted at the fastest rate when compared to other AuNPs. In the findings, a surface‐dependent biodistribution and elimination of AuNPs are indicated in silkworms, and the feasibility of using this inexpensive animal model for time‐ and cost‐effective, preliminary in vivo studies of NPs is confirmed.

Pediococcus pentosaceus ZZ61 enhances growth performance and pathogenic resistance of silkworm Bombyx mori by regulating gut microbiota and metabolites

Probiotics have attracted considerable attention in animal husbandry due to their positive effect on animal growth and health. This study aimed to screen candidate probiotic strain promoting the growth and health of silkworm and reveal the potential mechanisms. A novel probiotic Pediococcus pentosaceus strain (ZZ61) substantially promoted body weight gain, feed efficiency, and silk yield. These effects were likely mediated by changes in the intestinal digestive enzyme activity and nutrient provisioning (e.g., B vitamins) of the host, improving nutrient digestion and assimilation. Additionally, P. pentosaceus produced antimicrobial compounds and increased the antioxidant capacity to protect the host against pathogenic infection. Furthermore, P. pentosaceus affected the gut microbiome and altered the levels of gut metabolites (e.g., glycine and glycerophospholipids), which in turn promotes host nutrition and health. This study contributes to an improved understanding of the interactions between probiotic and host and promotes probiotic utilization in sericulture.

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability.

Encapsulation of single cells is a powerful technique used in various fields, such as regenerative medicine, drug delivery, tissue regeneration, cell-based therapies, and biotechnology. It offers a method to protect cells by providing cytocompatible coatings to strengthen cells against mechanical and environmental perturbations. Silk fibroin, derived from the silkworm Bombyx mori, is a promising protein biomaterial for cell encapsulation due to the cytocompatibility and capacity to maintain cell functionality. Here, THP-1 cells, a human leukemia monocytic cell line, were encapsulated with chemically modified silk polyelectrolytes through electrostatic layer-by-layer deposition. The effectiveness of the silk nanocoating was assessed using scanning electron microscopy (SEM) and confocal microscopy and on cell viability and proliferation by Alamar Blue assay and live/dead staining. An analysis of the mechanical properties of the encapsulated cells was conducted using atomic force microscopy nanoindentation to measure elasticity maps and cellular stiffness. After the cells were encapsulated in silk, an increase in their stiffness was observed. Based on this observation, we developed a mechanical predictive model to estimate the variations in stiffness in relation to the thickness of the coating. By tuning the cellular assembly and biomechanics, these encapsulations promote systems that protect cells during biomaterial deposition or processing in general.

Estrogen-related receptor, a molecular target against lepidoptera pests

Until recently, chemical pesticides were one of the most effective means of controlling agricultural pests; therefore, the search for insecticide targets for agricultural pests has been an ongoing problem. Estrogen-related receptors (ERRs) are transcription factors that regulate cellular metabolism and energy homeostasis in animals. Silkworms are highly sensitive to chemical pesticides, making them ideal models for pesticide screening and evaluation. In this study, we detected ERR expression in key organs involved in pesticide metabolism in silkworms (Bombyx mori), including the fat body and midgut. Using ChIP-seq technology, many estrogen- related response elements were identified in the 2000-bp promoter region upstream of metabolism-related genes, almost all of which were potential ERR target genes. The ERR inhibitor, XCT-790, and the endocrine disruptor, bisphenol A, significantly inhibited expression of the ERR target genes, BmTreh-1, BmTret-1, BmPK, BmPFK, and BmHK, in the fat bodies of silkworms, resulting in pupation difficulties in silkworm larvae that ultimately lead to death. In addition, based on the clarification that the ERR can bind to XCT-790, as observed through biofilm interferometry, its three-dimensional spatial structure was predicted, and using molecular docking techniques, small-molecule compounds with a stronger affinity for the ERR were identified. In summary, utilizing the powerful metabolic regulatory function of ERR in Lepidoptera pests, the developed small molecule inhibitors of ERR can be used for future control of Lepidoptera pests.

An In-Depth Insight into the Profile, Mechanisms, Functions, and Transfer of Essential Amino Acids from Mulberry Leaves to Silkworm Bombyx mori L. Pupae and Fish.

The silkworm Bombyx mori, the second most varied group of insects, is a fascinating insect that belongs to the Lepidoptera species. We aimed to deepen our knowledge about the composition and significance of amino acids (AA) from the sericulture chain to fish. AAs are the most prevalent molecules throughout the growth process of silkworms. We described AAs classification, occurrence, metabolism, and functions. Online datasets revealed that the essential AAs (EAA) level in fish meal and silkworm pupae (SWP) is comparable. SWP have a high content of methionine and lysine, which are the principal limiting AAs in fish diets, indicating that SWP have nutritional potential to be added to fish diets. Additionally, an overview of the data analyzed displays that SWP have a higher protein efficiency ratio than fish meal, the classical protein-rich source (>1.19 times), and compared to soybean meal, the second-most preferred source of protein in aquaculture (>2.08 times), indicating that SWP can be considered effective for animal feeding. In this study, we provide an overview of the current knowledge concerning AAs, paying special emphasis to EAAs and explaining, to some extent, certain mechanisms and functions of these compounds, from mulberry leaves to larvae-pupae and fish diets.

Biochemical characterization of Bombyx mori Dicer-2 that dices double-stranded RNAs into 20-nt small RNA.

We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (Bombyx mori) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from B. mori Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5'-phosphorylated dsRNAs at the 20-nt site-counted from the 5'-phosphorylated end-and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells.

Silk-Based 3D Porous Scaffolds for Tissue Engineering.

Silk fibroin, extracted from the silk of the Bombyx mori silkworm, stands out as a biomaterial due to its nontoxic nature, excellent biocompatibility, and adjustable biodegradability. Porous scaffolds, a type of biomaterial, are crucial for creating an optimal microenvironment that supports cell adhesion and proliferation, thereby playing an essential role in tissue remodeling and repair. Therefore, this review focuses on 3D porous silk fibroin-based scaffolds, first summarizing their preparation methods and then detailing their regenerative effects on bone, cartilage, tendon, vascular, neural, skin, hepatic, and tracheal epithelial tissue engineering in recent years.

Dietary inclusion of defatted silkworm (Bombyx mori L.) pupa meal in broiler chickens: phase feeding effects on nutritional and sensory meat quality

The present experiment was conducted to test the effect of a 4% defatted silkworm (Bombyx mori) pupae meal (SWM) incorporation into chickens’ diets at different growth phases on meat quality characteristics and sensory traits. Ninety ROSS 308 day-old male broiler chickens were randomly assigned to three dietary groups, with five replicated pens/diet: the first group received a control (C) diet throughout the growing period of 42 days, the second group received a diet with 4% SWM (SWM1) during the starter phase (1–10 days) and the C diet up to slaughter, whereas the third group was fed the C diet during the starter phase and 4% SWM during the grower and finisher phases (SWM2). Diets were isonitrogenous and isoenergy, and birds had free access to feed and water throughout the experimental trial. At 42 days of age, 15 chickens/treatment were slaughtered at a commercial abattoir. Fatty acid (FA) and amino acid (AA) profiles and contents, as well as oxidative status, were determined in both breast and leg meat cuts. Also, a descriptive sensory analysis was performed on breast meat by trained panelists. Results highlighted that the SWM2 treatment increased the n-3 proportion and content in both breast or leg meat, thereby improving the omega-6/omega-3 (n-6/n-3) ratio in both cuts (P<0.001). However, the dietary treatment had no significant effect on the oxidative status of either breast or leg meat (P>0.05). The SWM had a limited impact on overall sensory traits of breast meat, but it did contribute to improved meat tenderness in SWM-fed chickens (P<0.01). Furthermore, SWM1 meat exhibited higher juiciness (P<0.05) and off flavor intensity (P<0.05) compared to the control meat. Overall, the present experiment indicated that defatted SWM holds promise as an alternative ingredient in chicken rations, ensuring satisfactory meat quality. Furthermore, administering SWM during the grower-finisher phase demonstrated beneficial effects on meat healthiness, ultimately enhancing n-3 fatty acids content and reducing the n-6/n-3 ratio.

Upf2-Mediated Nonsense-Mediated Degradation Pathway Involved in Genetic Compensation of TrpA1 Knockout Mutant Silkworm (Bombyx mori).

Genetic mutations leading to premature termination codons are known to have detrimental effects. Using the Lepidoptera model insect, the silkworm (Bombyx mori), we explored the genetic compensatory response triggered by mutations with premature termination codons. Additionally, we delved into the molecular mechanisms associated with the nonsense-mediated mRNA degradation pathway. CRISPR/Cas9 technology was utilized to generate a homozygous bivoltine silkworm line BmTrpA1-/- with a premature termination. Transcript levels were assessed for the BmTrpA paralogs, BmPyrexia and BmPainless as well as for the essential factors Upf1, Upf2, and Upf3a involved in the nonsense-mediated mRNA degradation (NMD) pathway. Upf2 was specifically knocked down via RNA interference at the embryonic stage. The results comfirmed that the BmTrpA1 transcripts with a 2-base deletion generating a premature termination codon in the BmTrpA1-/- line. From day 6 of embryonic development, the mRNA levels of BmPyrexia, BmPainless, Upf1, and Upf2 were significantly elevated in the gene-edited line. Embryonic knockdown of Upf2 resulted in the suppression of the genetic compensation response in the mutant. As a result, the offspring silkworm eggs were able to hatch normally after 10 days of incubation, displaying a non-diapause phenotype. It was observed that a genetic compensation response does exist in BmTrpA1-/-B. mori. This study presents a novel discovery of the NMD-mediated genetic compensation response in B. mori. The findings offer new insights into understanding the genetic compensation response and exploring the gene functions in lepidopteran insects, such as silkworms.

Gut Microbiota Analysis in Silkworms (Bombyx mori) Provides Insights into Identifying Key Bacterials for Inclusion in Artificial Diet Formulations.

The gut microbiome significantly influences the health and productivity of silkworms (Bombyx mori), the cornerstone of sericulture. With the increasing use of cost-effective artificial diets in sericulture, it is crucial to understand how these diets impact the silkworm gut microbiomes. Here we employed 16S rRNA sequencing to delineate the impact of three distinct dietary regimens on the silkworm gut microbiomes: exclusive mulberry leaf diet (SY), exclusive artificial feed diet (SL), and a sequential transition from artificial feed to mulberry leaves (ZS). Our results unveiled stark differences in microbial diversity across the groups, with the ZS group displaying an intermediary complexity. LefSe and random forest analyses identified Methylobacteriaceae, Microbacterium, and Rhodococcus as significantly enriched in the ZS group, suggesting their potential to facilitate silkworms' adaptation to dietary transitions. Functional profiling revealed differential pathway regulation, indicating a metabolic reconfiguration in response to dietary modulations. Notably, the enrichment of Lactobacillus and Weissella in both the SL and ZS groups highlights their potential as probiotics in artificial diets. Our findings provide insights into the diet adaptation mechanisms of silkworm gut microbiota, paving the way for harnessing the intestinal bacteria to enhance silkworm health and silk production through targeted microbial interventions in sericulture practices.