Lipoplex-based RNA delivery system enhances RNAi efficiency for targeted Pest control in Spodoptera litura.

RNA interference (RNAi) represents a promising biotechnological strategy for sustainable pest control by silencing essential insect genes, resulting in increased mortality. However, its practical application remains limited due to environmental instability and delivery inefficiency of RNA molecules. In this study, three novel RNA-delivery nanoparticles-Polyplex, CS-TPP, and Lipoplex- were developed and their ability to enhance RNA stability and gene silencing efficacy against the intestinal mesh gene in Spodoptera litura was evaluated.

Differential RNAi efficacy of siRNA and dsRNA targeting key genes for pest control in .

RNA interference (RNAi) is a promising gene-silencing technique for pest control that targets essential genes. We assessed the potential of double-stranded RNA (dsRNA) and small interfering RNA (siRNA) to silence or genes in the midguts of larvae. Despite the theoretical promise of RNAi approaches, our findings revealed that dsRNA did not induce significant gene silencing or impact larval growth, whereas siRNA exhibited clear insecticidal effects, likely by disrupting intestinal osmoregulation and impairing larval fitness.

A Novel Chitin-Based Purification System Using GAL1 Fusion Tags: Enhancing Recombinant Protein Production While Retaining Biological Activity.

Efficient and economical purification methods are crucial for the commercial production of recombinant proteins with biomedical applications. In this study, we developed an affinity chromatography system that leverages the polysaccharide-binding properties of galectin-1 (GAL1) as a protein tag. The known GAL1-binding material, chitin, was used as the purification matrix.

Synergistic impacts of propargite exposure and deformed wing virus infection on the health of western honey bees.

This study aimed to elucidate the possible synergistic effects of chemical pesticides and viral infections. Our experiments demonstrated that the Varroa mite-borne deformed wing virus (DWV) by itself had a minimal impact on bees. Conversely, when bees were simultaneously treated with acaricides, their mortality rate increased.

A salivary GMC oxidoreductase of Manduca sexta re-arranges the green leaf volatile profile of its host plant.

Green leaf volatiles (GLVs) are short-chain oxylipins that are emitted from plants in response to stress. Previous studies have shown that oral secretions (OS) of the tobacco hornworm Manduca sexta, introduced into plant wounds during feeding, catalyze the re-arrangement of GLVs from Z-3- to E-2-isomers. This change in the volatile signal however is bittersweet for the insect as it can be used by their natural enemies, as a prey location cue.

Honey bee foraging ability suppressed by imidacloprid can be ameliorated by adding adenosine.

Honey bees are important pollinators in most ecosystem, but they are currently facing many threats, which have led to a reduction in their population. Previous studies have indicated that neonicotinoid pesticide can impair the memory and learning ability of honey bees, which can eventually lead to a decline in their foraging and homing abilities. In this study, we investigated the homing ability barrier from the perspective of energy supply.

Identification of silk components in the bombycoid moth Andraca theae (Endromidae) reveals three fibroin subunits resembling those of Bombycidae and Sphingidae.

The silk produced by Lepidoptera caterpillars is a mixture of proteins secreted by the transformed labial glands, the silk glands (SG). The silk fiber consists of insoluble filamentous proteins that form a silk core and are produced in the posterior part of the SG and soluble coat proteins consisting of sericins and various other polypeptides secreted in the middle part of the SG. We constructed a silk gland specific transcriptome of Andraca theae and created a protein database required for peptide mass fingerprinting.

Elevated temperature affects energy metabolism and behavior of bumblebees.

Bumblebees (Bombus eximius) are one of the most prominent pollinators in the agricultural industry because of their adaptation to temperate climates and pollination behavior (buzz pollination). Several studies have explained the need to increase conservation efforts for bumblebees due to climate change, but studies on the impact of climate change on pollination behavior of bumblebees have been limited. The present study investigated the effect of elevated temperatures on the survival and physiology of bumblebees.

Enhancing the insecticidal potential of a baculovirus by overexpressing the mammalian β-galactosyl binding protein galectin-1.

Background: Bio-pesticide development is an important area of research in agriculture, in which viruses are an essential tool. Infection by entomological pathogenic viruses kills agricultural pests, and viral progenies are disseminated to infect more pests, eventually achieving long-term pest control in the field. Of the current virus-based pest control models, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most studied.

Carbohydrate metabolism is a determinant for the host specificity of baculovirus infections.

Baculoviruses Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV) have highly similar genome sequences but exhibit no overlap in their host range. After baculovirus infects nonpermissive larvae (e.g.

Real-time monitoring of deformed wing virus-infected bee foraging behavior following histone deacetylase inhibitor treatment.

Impairment in the learning/memory behavior of bees is responsible for the massive disappearance of bee populations and its consequent agricultural economic losses. Such impairment might be because of o both pesticide exposure and pathogen infection, with a key contributor deformed wing virus (DWV). The present study found that sodium butyrate (NaB) significantly increased survival and reversed the learning/memory impairment of DWV-infected bees.

Deformed wing virus infection affects the neurological function of Apis mellifera by altering extracellular adenosine signaling.

Deformed wing virus (DWV) infection is believed to be closely associated with colony losses of honeybee (Apis mellifera) due to reduced learning and memory of infected bees. The adenosine (Ado) pathway is important for maintaining immunity and memory function in animals, and it enhances antivirus responses by regulating carbohydrate metabolism in insects. Nevertheless, its effect on the memory of invertebrates is not yet clear.

MicroRNAs from Snellenius manilae bracovirus regulate innate and cellular immune responses of its host Spodoptera litura.

To avoid inducing immune and physiological responses in insect hosts, parasitoid wasps have developed several mechanisms to inhibit them during parasitism, including the production of venom, specialized wasp cells, and symbioses with polydnaviruses (PDVs). These mechanisms alter the host physiology to give the wasp offspring a greater chance of survival. However, the molecular mechanisms for most of these alterations remain unclear.

Identification of Immune Regulatory Genes in through Caffeine Treatment.

Plants and pollinators are mutually beneficial: plants provide nectar as a food source and in return their pollen is disseminated by pollinators such as honeybees. Some plants secrete chemicals to deter herbivores as a protective measure, among which is caffeine, a naturally occurring, bitter tasting, and pharmacologically active secondary compound. It can be found in low concentrations in the nectars of some plants and as such, when pollinators consume nectar, they also take in small amounts of caffeine.

Adenosine Receptor Modulates Permissiveness of Baculovirus (Budded Virus) Infection via Regulation of Energy Metabolism in .

Although the modulation of host physiology has been interpreted as an essential process supporting baculovirus propagation, the requirement of energy supply for host antivirus reactions could not be ruled out. Our present study showed that metabolic induction upon AcMNPV (budded virus) infection of stimulated virus clearance and production of the antivirus protein, gloverin. In addition, we demonstrated that adenosine receptor signaling (AdoR) played an important role in regulating such metabolic reprogramming upon baculovirus infection.

Snellenius manilae bracovirus suppresses the host immune system by regulating extracellular adenosine levels in Spodoptera litura.

Sufficient energy supply to the host immune system is important for resisting pathogens. Therefore, during pathogen infection, the host metabolism is reassigned from storage, growth, and development to the immune system. Previous studies in Drosophila melanogaster have demonstrated that systemic metabolic switching upon an immune challenge is activated by extracellular adenosine signaling, modulating carbohydrate mobilization and redistributing energy to the hemocytes.

Identification of Regulatory Host Genes Involved in Sigma Virus Replication Using RNAi Knockdown in .

The sigma virus, a member of the family, specifically propagates itself in . It contains six genes in the order of 3'-------5'. The sigma virus is the only arthropod-specific virus of the family.

MicroRNAs derived from the insect virus HzNV-1 promote lytic infection by suppressing histone methylation.

Heliothis zea nudivirus-1 (HzNV-1) is an insect virus that can induce both lytic and latent infections in various insect cell lines. During latent infection, several microRNAs (miRNAs) are produced from persistency-associated gene 1 (pag1) as the only detectable HzNV-1 transcript. Previous studies have shown that the pag1 gene suppresses the immediate-early gene hhi1 and promotes host switching into a latent infection via miRNAs derived from pag1.

Regulation of genes related to immune signaling and detoxification in Apis mellifera by an inhibitor of histone deacetylation.

The western honeybee (Apis mellifera) is essential for the global economy due to its important role in ecosystems and agriculture as a pollinator of numerous flowering plants and crops. Pesticide abuse has greatly impacted honeybees and caused tremendous loss of honeybee colonies worldwide. The reasons for colony loss remain unclear, but involvement of pesticides and pathogen-pesticide interactions has been hypothesized.

Identification of a high-efficiency baculovirus DNA replication origin that functions in insect and mammalian cells.

Unlabelled: The p143 gene from Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) has been found to increase the expression of luciferase, which is driven by the polyhedrin gene promoter, in a plasmid with virus coinfection. Further study indicated that this is due to the presence of a replication origin (ori) in the coding region of this gene. Transient DNA replication assays showed that a specific fragment of the p143 coding sequence, p143-3, underwent virus-dependent DNA replication in Spodoptera frugiperda IPLB-Sf-21 (Sf-21) cells.

A non-coding RNA of insect HzNV-1 virus establishes latent viral infection through microRNA.

Heliothis zea nudivirus-1 (HzNV-1) is an insect virus previously known as Hz-1 baculovirus. One of its major early genes, hhi1, is responsible for the establishment of productive viral infection; another gene, pag1, which expresses a non-coding RNA, is the only viral transcript detectable during viral latency. Here we showed that this non-coding RNA was further processed into at least two distinct miRNAs, which targeted and degraded hhi1 transcript.

Heliothis zea nudivirus 1 gene hhi1 induces apoptosis which is blocked by the Hz-iap2 gene and a noncoding gene, pag1.

Heliothis zea nudivirus 1 (HzNV-1 or Hz-1 virus), previously regarded as a nonoccluded baculovirus, recently has been placed in the Nudivirus genus. This virus generates HzNV-1 HindIII-I 1 (hhi1) and many other transcripts during productive viral infection; during latent viral infection, however, persistency-associated gene 1 (pag1) is the only gene expressed. In this report, we used transient expression assays to show that hhi1 can trigger strong apoptosis in transfected cells, which can be blocked, at least partially, by the inhibitor of apoptosis genes Autographa californica iap2 (Ac-iap2) and H.

Autographa californica multiple nucleopolyhedrovirus LEF-2 is a capsid protein required for amplification but not initiation of viral DNA replication.

The late expression factor 2 gene (lef-2) of baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) has been identified as one of the factors essential for origin-dependent DNA replication in transient expression assays and has been shown to be involved in late/very late gene expression. To study the function of lef-2 in the life cycle of AcMNPV, lef-2 knockout and repair bacmids were generated by homologous recombination in Escherichia coli. Growth curve analysis showed that lef-2 was essential for virus production.

The early gene hhi1 reactivates Heliothis zea nudivirus 1 in latently infected cells.

Heliothis zea nudivirus 1 (HzNV-1), previously known as Hz-1 virus, is an insect virus able to establish both productive and latent infections in several lepidopteran insect cells. Here, we have cloned and characterized one of the HzNV-1 early genes, hhi1, which maps to the HindIII-I fragment of the viral genome. During the productive viral infection, a 6.

The establishment of a controllable expression system in baculovirus: stimulated overexpression of polyhedrin promoter by LEF-2.

Previously, controllable gene expression in baculovirus was not possible using an insect system. We found that this was due to a high background activation of minimal promoter by the viral polyhedrin upstream (pu) sequence. Here, by truncation of the pu sequence, regulatory gene expression was established through the tetracycline regulatory expression system.