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Abstract Background Booklice, belonging to the genus Liposcelis (Psocodea: Liposcelididae), commonly known as psocids, infest a wide range of stored products and are implicated in the transmission of harmful microorganisms such as fungi and bacteria. The olfactory system is critical for insect feeding and reproduction. Elucidating the molecular mechanisms of the olfactory system in booklice is crucial for developing effective control strategies. In this study, we aim to bridge this knowledge gap by leveraging the transcriptome and genome data from five Liposcelis species. Result Using HMMER method and manual annotation, we have identified common gene families associated with olfactory processes, including odorant binding proteins (OBPs), chemosensory proteins (CSPs), odorant receptors (ORs), ionotropic receptors (IRs), and sensory neuron membrane proteins (SNMPs). Specifically, we identified 94, 118, 26, 47, and 34 olfactory-related genes in L. bostrychophila, L. tricolor, L. entomophila, L. decolor, and L. yangi, respectively. Comparison of quantities revealed that the number of ORs and IRs in the genome is significantly higher than those identified in the transcriptome. This discrepancy may be due to the specific expression of these genes in certain tissues or their lack of expression during the experimental stage. Simultaneously, analysis of gene expression profiles across different developmental stages revealed varying periods of peak expression for olfactory-related genes. These results suggest that the identification of olfactory-related genes in booklice on a genome-wide scale is more feasible and reliable than using a transcriptome-based approach. Additionally, compared to parasitic lice, booklice possess significantly more olfactory-related genes. This increase may be due to the inability of parasitic lice to survive without a host, whereas booklice have a wide range of feeding habits and live in complex and variable environments. Furthermore, we observed that the IR gene family in L. tricolor has undergone a certain degree of amplification, which may facilitate its adaptation to diverse environmental conditions. Conclusions We identified olfactory-related genes of five Liposcelis species for the first time, providing valuable insights for future functional investigations into olfactory genes associated with pheromone and odorant recognition. These discoveries present promising targets for effectively managing psocid pests.

Abstract Liposcelis tricolor (Psocoptera: Liposcelididae) is a significant pest affecting stored products globally. However, due to the lack of a detailed genomic reference, the mechanisms of sex determination, stress resistance, and potential control methods for this booklouse remain poorly understood. In this study, the chromosome-level genome of L. tricolor was assembled by employing Illumina, Nanopore, and Hi-C sequencing technologies. The final genome size was determined to be 229.33 Mb, anchored to 9 pseudo-chromosomes. BUSCO analysis showed that 99.2% of complete BUSCOs were identified, suggesting the high completeness of the genome. A total of 91.49 Mb of repetitive sequences, accounting for 38.84% of the total genome, were annotated, and 15,647 protein-coding genes were predicted, with 88.17% functionally annotated. Additionally, we identified 25 typical sex-determining genes based on the genomic data. This high-quality genome assembly provides a crucial foundation for advancing our comprehension of the molecular biology, genetics, and potential control strategies for psocid L. tricolor.

Sophora davidii is a cross-pollinated plant with important ecological protection and medicinal value in China, but its seed yield is low due to backward and nonstandard production technology. Therefore, we divide the flowering period of Sophora davidii into initial, full and final flowering period, measuring the floral morphology, pollen viability, stigma receptivity, nectar volume and nectar concentration, foraging behavior of pollinators, fertilization physiology, seed yield and quality through field observation and indoor testing to explore whether the flowering period affects the floral traits, pollinator behavior and seed production of plants. Our results revealed that the nectar volume, nectar concentration, pollen viability and stigma receptivity at full flowering period were the highest. The single visit time and visit time per flower of Chinese honey bees were the longest in the full flowering period, while the number of transfer, visit frequency and number of touching stigma were the least. The visiting number of the bees was the most and the most active in the full flowering period. The bees pollination not only improved the pollen amount, germination rate, pollen tube length and the ovule number of S. davidii, but also their effect was the most obvious in full flowering period. The principal component analysis showed that pollination by Chinese honey bees during the whole flowering period of S. davidii was the best way to produce seeds. We can conclude that flowering period affects flower traits, foraging behavior of pollinators, seed yield and quality of S. davidii.

Abstract Natalisin (NTL) is a conserved neuropeptide, only present in insects, that has been reported to regulate their sexual activity. In this study, we investigated the involvement of NTL in the reproductive behaviors of a major invasive pest, Spodoptera frugiperda. We identified NTL precursor-encoded transcripts, and evaluated their transcript levels in different stages and tissues of S. frugiperda. The results showed that the NTL transcript level was expressed in both male and female pupae and both male and female adults in the later stage. It was highly expressed in male pupae, 3-day-old male and female adults, and 5-day-old male adults. In different tissues, the expression level is higher in the male and female adult brain and male testis. Immunohistochemical staining of the brain of S. frugiperda female and male adults revealed that three pairs of brain neurons of S. frugiperda adults of both sexes secreted and expressed NTL. To study the role of NTL in reproductive behaviors, NTL was silenced in S. frugiperda male and female adults by RNA interference (RNAi) technology, the results showed that silencing NTL could significantly affect the sexual activity behavior of the adults, reducing the calling rate of females, the courtship rate of males, and the mating rate. In summary, this study emphasizes the important role of NTL in regulating the mating behavior and sexual activity of S. frugiperda in both male and female adults, potentially laying a foundation to employ NTL as a new insect-specific target to control populations of pest insects.

Abstract The pink stem borer, Sesamia inferens Walker (Lepidoptera: Noctuidae), is one of the most notorious pest insects of rice and maize crops in the world. Here, we generated a high-quality chromosome-level genome assembly of S. inferens, using a combination of Illumina, PacBio HiFi and Hi-C technologies. The total assembly size was 973.18 Mb with a contig N50 of 33.39 Mb, anchored to 31 chromosomes, revealing a karyotype of 30 + Z. The BUSCO analysis indicated a high completeness of 98.90% (n = 5286), including 5172 (97.8%) single-copy BUSCOs and 58 (1.1%) duplicated BUSCOs. The genome contains 58.59% (564.58 Mb) repeat elements and 26628 predicted protein-coding genes. The chromosome-level genome assembly of S. inferens provides in-depth knowledge and will be a helpful resource for the Lepidoptera and pest control research communities.

Ultraviolet B (UV-B) is a significant environmental factor affecting insect development, survival, and reproduction. DNAJ proteins are molecular chaperones found ubiquitously in insects that are crucial for their adaptation to environmental stresses. This study aimed to elucidate the roles of DNAJ genes in the response of Myzus persicae to UV-B stress. Herein, we identified six DNAJ genes in the aphid M. persicae, a devastating agricultural pest. We analyzed their expression profiles at different stages of development, in different tissues and for various durations of UV-B exposure. The expression levels of MpDNAJC30, MpDNAJC11, MpDNAJC2, and MpDNAJC3 were highest in wingless adults, while MpDNAJC9 and MpDNAJC17 were highest in second- and third-instar nymphs, respectively. Six MpDNAJs had higher expression levels in the epidermis and embryos, and lower levels in the head. Additionally, the expression levels of all genes increased significantly under different durations of UV-B exposure. Knockout of the DNAJ genes using RNA interference caused a significant decline in the survival rate, weight, body length, and body width of M. persicae exposed to UV-B radiation. Our research provides valuable insights into the stress response mechanisms of M. persicae, highlighting the importance of DNAJ genes in mediating their adaptation to UV-B stress.

Dr. Pieter Oomen, born in 1946, a prominent and inspirational scientist in many areas of the biosafety of pesticides, most notably ecotoxicology, regulatory, risk assessment and bioassays, passed away peacefully on 19th of January 2024. His efforts have had a great and long-lasting impact on bee risk assessment and have improved bee protection. While we will miss a very warm and brilliant person; his many contributions to science and our ability to detect and mitigate risks to the environment remain.

Abstract Macropsini is a tribe of Eurymelinae in the family Cicadellidae that is widely distributed worldwide. Still, its taxonomic status has been unstable, and the classification of certain clades at the genus level has been controversial. The aim of this study is to address the patterns and processes that explain the structure and the evolution of the mitogenomes of Macropsini, while contributing to the resolution of systematic issues involving five of their genera. To this task, the mitogenomes of 26 species of the tribe were sequenced and characterized, and their phylogenetic relationships were reconstructed. The results revealed that the nucleotide composition of mitochondrial genes in these 26 species was significantly skewed toward A and T. Codons ending with T or A in relative synonymous codon usage were significantly more prevalent than those ending with C or G. The parity plot, neutrality plot, and correspondence analysis revealed that mutation and selective pressure affect codon usage patterns. In the phylogenetic relationships of the Macropsini, the monophyly of Pedionis and Macropsis was well‐supported. Meanwhile, Oncopsis revealed paraphyletic regarding Pediopsoides. In conclusion, this research not only contributes the valuable data to the understanding of the mitogenome of the Macropsini but also provides a reference for future investigations on codon usage patterns, potential adaptive evolution, and the phylogeny of the mitogenome within the subfamily Eurymelinae.

Occurrences of introduced and invasive pollinators are increasing worldwide. To predict the potential impacts of exotic bees on native ecosystems we need to understand their ecological interactions. Life history traits are fundamental for understanding ecological interactions and often help to explain the spread of exotic species. We conducted home-and-away comparisons of life history traits of the first invasive bee in Europe, the sculptured resin bee Megachile sculpturalis Smith 1853. We compiled information on nest architecture, offspring, natural enemies, body size and phenology using published literature, museum specimen, data from citizen science initiatives, field observations and reared specimen. Megachile sculpturalis uses a broad variety of nesting materials for brood cell construction, including even plastic at the exotic range. Body size at warm temperate climate was similar in the native and exotic ranges, but phenology shifted forward by about one month (28.9 ± 3.3 SE days) in the exotic ranges. The abundance of natural enemies was similar between native and exotic ranges but specialist enemies were missing in the exotic ranges. These trait shifts may be explained by founder effects or ecological filtering. The comparison of life history traits in native and exotic ranges sheds light on the ecological-evolutionary process of this quickly spreading species and provides a better understanding of invasion processes in solitary bees.

Abstract Elucidating the intricate interactions between viral pathogens and host cellular machinery during infection is paramount for understanding pathogenic mechanisms and identifying potential therapeutic targets. The RNA modification N 6-methyladenosine (m6A) has emerged as a significant factor influencing the trajectory of viral infections. Hence, the precise and quantitative mapping of m6A modifications in both host and viral RNA is pivotal to understanding its role during viral infection. With the rapid advancement of sequencing technologies, scientists are able to detect m6A modifications with various quantitative, high-resolution, transcriptome approaches. These technological strides have reignited research interest in m6A, underscoring its significance and prompting a deeper investigation into its dynamics during viral infections. This review provides a comprehensive overview of the historical evolution of m6A epitranscriptome sequencing technologies, highlights the latest developments in transcriptome-wide m6A mapping, and emphasizes the innovative technologies for detecting m6A modification. We further discuss the implications of these technologies for future research into the role of m6A in viral infections. Graphical Abstract

Mosquito control still relies heavily on synthetic molecules, which can lead to the selection of resistant populations and undesirable environmental problems. This study described the preparation of a nanoparticle of the plant-derived molecule, β-myrcene, with chitosan, and the assessment of its toxicity against larvae of the yellow fever mosquito, Aedes aegypti. By producing fluorescent chitosan nanoparticles, we were able to observe their distribution in the digestive tract of larvae of Ae. aegypti. Chitosan-based nanoparticles containing β-myrcene (238 mg/L) could kill 100% of the larvae tested, whereas the blank control (i.e., the nanoparticle without β-myrcene) showed no larvicidal activity. The chitosan nanoparticles with β-myrcene had a zeta potential of +15 mV and a hydrodynamic diameter ranging from 30 to 2800 nm. The blank control, without β-myrcene, had a zeta potential of +26 mV and a diameter of 30 to 830 nm. Fluorescence analysis showed that the nanoparticles were efficiently absorbed and distributed in the digestive tract organs of the Ae. aegypti larvae. In short, our results reinforce the benefits of using chitosan to carry molecules of plant-derived-molecules, such as β-myrcene, in mosquito control, suggesting a broad internal distribution that contributes to their toxicity.

Neuropeptide CCHamide-1 (abbreviated as CCHa1) is a recently discovered peptide that is present in many arthropods and is the ligand of the CCHa1R, a member of the G protein-coupled receptors (GPCRs) superfamily, which plays a regulatory role in diverse physiological processes such as feeding, circadian rhythm, insulin production, lipid metabolism, growth, and reproduction. However, the function of this gene in aphids is still unknown. Here, we characterized and determined the potential role of CCHa1/CCHa1R signaling in the pea aphid, Acyrthosiphon pisum, which is a notorious pest in agriculture. The docking analysis revealed that the CCHa1 peptide binds to its receptor CCHa1R through specific amino acid residues, which are critical for maintaining the structural and functional integrity of the peptide–receptor complex. Quantitative real-time reverse transcription-PCR (qRT-PCR) revealed the expression levels of CCHa1/CCHa1R transcripts in different development stages and different tissues, indicating that the CCHa1 expression was high in the first nymphal instar compared to the upcoming nymphal instars and adults, and was predominantly high in the brain. The CCHa1/CCHa1R transcript levels were significantly upregulated in starved aphids compared to fed aphids. Moreover, RNAi knockdown by the injection of dsRNA-CCHa1 and dsRNA-CCHa1R significantly reduced the corresponding expression of the target gene and reduced their food intake in adult aphids, as revealed by the electrical penetration graph results. CCHa1/CCHa1R-silencing also reduced the reproduction, but not the survival, in A. pisum. Our data demonstrated that CCHa1/CCHa1R play a role in the regulation of feeding in A. pisum, suggesting a role of the CCHa1 signaling pathway in the aphids relating to their nutritional status.

Abstract Background Pink stem borer, Sesamia inferens (Walker), a major rice pest, has rarely been reported to cause serious damage to sorghum in China. During a survey of insect pests in glutinous sorghum for Chinese liquor production areas in Guizhou Province, it was found that S. inferens has caused serious damage to local sorghum in recent years. Methods In this context, we assessed the adaptation of S. inferens to three gramineous crop host plants (maize, rice and sorghum) based on age-stage, two-sex life table theory. Results Sesamia inferens had the highest development time, survival rate and total longevity on rice, followed by on sorghum and maize. There was no significant difference in intrinsic rate of increase and finite rate of increase among maize (r = 0.0702 day−1, λ = 1.0727 day−1), sorghum (r = 0.0681 day−1, λ = 1.0705 day−1) and rice (r = 0.0645 day−1, λ = 1.0666 day−1), but the mean generation period (T) was significantly higher for populations reared on rice (53.84 days) than on sorghum (49.53 days) and maize (47.31 days). Conclusions The results of our study indicate that S. inferens was able to complete the full developmental cycle on all three host plants. This study further supports that glutinous sorghum is an adaptable host plant for S. inferens.

Abstract Background To understand mechanisms of adaptation and plasticity of pollinators and other insects a better understanding of diversity and function of their key symbionts is required. Commensalibacter is a genus of acetic acid bacterial symbionts in the gut of honey bees and other insect species, yet little information is available on the diversity and function of Commensalibacter bacteria. In the present study, whole-genome sequences of 12 Commensalibacter isolates from bumble bees, butterflies, Asian hornets and rowan berries were determined, and publicly available genome assemblies of 14 Commensalibacter strains were used in a phylogenomic and comparative genomic analysis. Results The phylogenomic analysis revealed that the 26 Commensalibacter isolates represented four species, i.e. Commensalibacter intestini and three novel species for which we propose the names Commensalibacter melissae sp. nov., Commensalibacter communis sp. nov. and Commensalibacter papalotli sp. nov. Comparative genomic analysis revealed that the four Commensalibacter species had similar genetic pathways for central metabolism characterized by a complete tricarboxylic acid cycle and pentose phosphate pathway, but their genomes differed in size, G + C content, amino acid metabolism and carbohydrate-utilizing enzymes. The reduced genome size, the large number of species-specific gene clusters, and the small number of gene clusters shared between C. melissae and other Commensalibacter species suggested a unique evolutionary process in C. melissae, the Western honey bee symbiont. Conclusion The genus Commensalibacter is a widely distributed insect symbiont that consists of multiple species, each contributing in a species specific manner to the physiology of the holobiont host.

AbstractCarbon quantum dots (CQDs) have many potential applications due to their cell-penetrating ability, biocompatibility and tunable properties. Among a variety of characteristics, the inhibition of bacteria by CQDs is often reported. However, the effect on other microorganisms, such as plant pathogenic fungi and oomycetes, is poorly studied. Here we monitored the growth of the oomycete plant pathogen Phytophthora infestans in the presence of CQDs, as well as of another three fungal plant pathogens, namely Botrytis cinerea, Alternaria alternata and Fusarium oxysporum. Moreover, the ability of CQDs to improve gene silencing caused by exogenous dsRNA in P. infestans was studied, and the toxicity of CQDs to human keratinocytes was evaluated. Our results indicate significant inhibitory activity of CQDs against P. infestans at relatively low concentrations. In a species-specific manner and to a lesser extent, the growth of the three fungal plant pathogens was also affected. We also found that the treatment of P. infestans with naked dsRNA in vitro did not trigger gene silencing. However, the mixture of CQDs with dsRNA increased RNAi efficiency, by causing a significant reduction of the transcript levels of the target gene in developing sporangia. Finally, no cytotoxicity of the CQDs, in the concentrations active against the plant pathogens, was found.

Abstract Body size is a trait that can affect plant–pollinator interaction efficiency and plant reproductive success. We explored the impact of intraspecific size shifts on the interactions between pollinators and flowering plants under controlled conditions. We considered two development conditions leading to the production of large and small individual flowers of Borago officinalis and Echium plantagineum. We also used the natural variability of worker size within bumblebee colonies to isolate small and large workers. We performed a fully crossed experiment with the two flower sizes of each plant species and the two sizes of bumblebee workers. Our results show that the size of both partners did not affect bee foraging behavior in most of the evaluated parameters and both bee sizes were equally efficient in depositing pollen. Significant differences were found only in pollen deposition across the life of a flower in small flowers of B. officinalis, with the greatest quantity of pollen deposited by small bees. We did not find a relationship between pollinator size and plant fitness. Our results suggest that generalist plant–pollinator interactions may be resilient to future potential mismatches in the size of the partners but remain to be tested if they are still resilient under the new environmental conditions resulting from global changes.

The alarming decline of pollinator populations has raised significant concerns worldwide and prompted the need for effective pesticide risk assessment within the Integrated Pest and Pollinator Management (IPPM) framework. This paper examines the diverse approaches to pollinator protection within the pesticide regulatory environments of the United States (US), the European Union (EU), and selected Asian countries. The US adopts a reactive approach, regulating pesticides only after evidence of harm emerges, while the EU embraces a proactive stance under the precautionary principle. The EU has implemented stringent regulations, including neonicotinoid bans, and conducts coordinated research on pesticide impacts. In contrast, some Asian countries face challenges with inadequate regulations, leading to adverse health and environmental consequences. This article highlights the need for comprehensive pesticide regulations across different regions to safeguard pollinators and mitigate the non-target risks associated with pesticide use.

Japanagallia is a genus of Cicadomorpha in the family of leafhoppers that are plant piercing-sucking insects, and it is difficult to distinguish by morphological characteristics. So far, only one complete mitochondrial genome data has been reported for the genus Japanagallia. Therefore, in order to better understand this group, we assembled and annotated the complete mitochondrial genomes of five Japanagallia species, and analyzed their codon usage patterns. Nucleotide composition analysis showed that AT content was higher than GC content, and the protein-coding sequences preferred to end with A/T at the third codon position. Relative synonymous codon usage analysis revealed most over-represented codon ends with A or T. Parity plot analysis revealed the codon usage bias of mitochondrial genes was influenced by both natural selection and mutation pressure. In the neutrality plot, the slopes of regression lines were < 0.5, suggesting that natural selection was playing a major role while mutation pressure was of minor importance. The effective number of codons showed that the codon usage bias between genes and genomes was low. Correspondence analysis revealed that the codon usage pattern differed among 13 protein-coding genes. Phylogenetic analyses based on three datasets using two methods (maximum likelihood and Bayesian inference), restored the Megophthalminae monophyly with high support values (bootstrap support values (BS) = 100, Bayesian posterior probability (PP) = 1). In the obtained topology, the seven Japanagallia species were clustered into a monophyletic group and formed a sister group with Durgade. In conclusion, our study can provide a reference for the future research on organism evolution, identification and phylogeny relationships of Japanagallia species.

Abstract The devastating Varroa mite (Varroa destructor Anderson and Trueman) is an obligatory ectoparasite of the honey bee, contributing to significant colony losses in North America and throughout the world. The limited number of conventional acaricides to reduce Varroa mites and prevent disease in honey bee colonies is challenged with wide-spread resistance and low target-site selectivity. Here, we propose a biorational approach using comparative genomics for the development of honey bee-safe and selective acaricides targeting the Varroa mite-specific neuropeptidergic system regulated by proctolin, which is lacking in the honey bee. Proctolin is a highly conserved pentapeptide RYLPT (Arg-Tyr-Leu-Pro-Thr) known to act through a G protein-coupled receptor to elicit myotropic activity in arthropod species. A total of 33 different peptidomimetic and peptide variants were tested on the Varroa mite proctolin receptor. Ligand docking model and mutagenesis studies revealed the importance of the core aromatic residue Tyr2 in the proctolin ligand. Peptidomimetics were observed to have significant oral toxicity leading to the paralysis and death of Varroa mites, while there were no negative effects observed for honey bees. We have demonstrated that a taxon-specific physiological target identified by advanced genomics information offers an opportunity to develop Varroa mite-selective acaricides, hence, expedited translational processes.

Nearly two fifths of the Earth's land area is currently used for agriculture, substantially impacting the environment and ecosystems. Besides the direct impact through land use change, intensive agriculture can also have an indirect impact, for example by changing wildlife epidemiology. We review here the potential effects of mass-flowering crops (MFCs), which are rapidly expanding in global cropping area, on the epidemiology of known pathogens in bee pollinators. We bring together the fifty MFCs with largest global area harvested and give an overview of their pollination dependency as well as their impact on bee pollinators. When in bloom these crops provide an abundance of flowers, which can provide nutrition for bees and increase bee reproduction. After their short bloom peak, however, the fields turn into green deserts. These big changes in floral availability strongly affect the plant-pollinator network, which in turn affects the pathogen transmission network, mediated by shared flowers. We address this dual role of flowers provided by MFCs, serving as nutritional resources as well as pathogen transmission spots, and bring together the current knowledge to assess how MFCs could affect pathogen prevalence in bee pollinator communities.

Varroa destructor is an ectoparasitic mite and is considered one of the most important causes of honey bee population loss. In the last years, substances of botanical origin have emerged as natural alternatives to diminish the mite population levels. Propolis is a natural product and is used by honey bees for multiple tasks, including protection from pathogens and parasites, and varroacidal activity of propolis extracts has been shown. In this study, we investigated the potential of propolis, collected by native Algerian honey bee subspecies (Apis mellifera intermissa and A. m. sahariensis) in different locations in Algeria and extracted by ultrasound, to control mites of V. destructor and tested the safety for the honey bees. The most important results were that the best propolis extracts at 10% killed 100% of the Varroa mites within 3–4 h in a Petri dish assay. In addition, when we sprayed A. m. intermissa bees infested with Varroa mites with a 10% concentration in a mini-hive setup, we scored a high mite mortality of 85–87% with the best propolis extracts, and importantly, there was no mortality in the bees. Our data demonstrated that propolis extracts in Algeria could be used in honey bee colonies by spraying against Varroa mite infestations, which may develop as an easy method for local beekeepers to control Varroa in their hives. Further research should investigate the mechanism of action.

Abstract Viruses are omnipresent, yet the knowledge on drivers of viral prevalence in wild host populations is often limited. Biotic factors, such as sympatric managed host species, as well as abiotic factors, such as climatic variables, are likely to impact viral prevalence. Managed and wild bees, which harbor several multi-host viruses with a mostly fecal–oral between-species transmission route, provide an excellent system with which to test for the impact of biotic and abiotic factors on viral prevalence in wild host populations. Here we show on a continental scale that the prevalence of three broad host viruses: the AKI-complex (Acute bee paralysis virus, Kashmir bee virus and Israeli acute paralysis virus), Deformed wing virus, and Slow bee paralysis virus in wild bee populations (bumble bees and solitary bees) is positively related to viral prevalence of sympatric honey bees as well as being impacted by climatic variables. The former highlights the need for good beekeeping practices, including Varroa destructor management to reduce honey bee viral infection and hive placement. Furthermore, we found that viral prevalence in wild bees is at its lowest at the extreme ends of both temperature and precipitation ranges. Under predicted climate change, the frequency of extremes in precipitation and temperature will continue to increase and may hence impact viral prevalence in wild bee communities.

Abstract In 1977, a sample of diseased adult honeybees (Apis mellifera) from Egypt was found to contain large amounts of a previously unknown virus, Egypt bee virus, which was subsequently shown to be serologically related to deformed wing virus (DWV). By sequencing the original isolate, we demonstrate that Egypt bee virus is in fact a fourth unique, major variant of DWV (DWV-D): more closely related to DWV-C than to either DWV-A or DWV-B. DWV-A and DWV-B are the most common DWV variants worldwide due to their close relationship and transmission by Varroa destructor. However, we could not find any trace of DWV-D in several hundred RNA sequencing libraries from a worldwide selection of honeybee, varroa and bumblebee samples. This means that DWV-D has either become extinct, been replaced by other DWV variants better adapted to varroa-mediated transmission, or persists only in a narrow geographic or host range, isolated from common bee and beekeeping trade routes.

Abstract Several plant lectins, or carbohydrate-binding proteins, interact with glycan moieties on the surface of immune cells, thereby influencing the immune response of these cells. Orysata, a mannose-binding lectin from rice, has been reported to exert immunomodulatory activities on insect cells. While the natural lectin is non-glycosylated, recombinant Orysata produced in the yeast Pichia pastoris (YOry) is modified with a hyper-mannosylated N-glycan. Since it is unclear whether this glycosylation can affect the YOry activity, non-glycosylated rOrysata was produced in Escherichia coli (BOry). In a comparative analysis, both recombinant Orysata proteins were tested for their carbohydrate specificity on a glycan array, followed by the investigation of the carbohydrate-dependent agglutination of red blood cells (RBCs) and the carbohydrate-independent immune responses in Drosophila melanogaster S2 cells. Although YOry and BOry showed a similar carbohydrate-binding profiles, lower concentration of BOry were sufficient for the agglutination of RBCs and BOry induced stronger immune responses in S2 cells. The data are discussed in relation to different hypotheses explaining the weaker responses of glycosylated YOry. In conclusion, these observations contribute to the understanding how post-translational modification can affect protein function, and provide guidance in the selection of the proper expression system for the recombinant production of lectins.

Human-induced environmental impacts on wildlife are widespread, causing major biodiversity losses. One major threat is agricultural intensification, typically characterised by large areas of monoculture, mechanical tillage, and the use of agrochemicals. Intensification leads to the fragmentation and loss of natural habitats, native vegetation, and nesting and breeding sites. Understanding the adaptability of insects to these changing environmental conditions is critical to predicting their survival. Bumblebees, key pollinators of wild and cultivated plants, are used as model species to assess insect adaptation to anthropogenic stressors. We investigated the effects of agricultural pressures on two common European bumblebees, Bombus pascuorum and B. lapidarius. Restriction-site Associated DNA Sequencing was used to identify loci under selective pressure across agricultural-natural gradients over 97 locations in Europe. 191 unique loci in B. pascuorum and 260 in B. lapidarius were identified as under selective pressure, and associated with agricultural stressors. Further investigation suggested several candidate proteins including several neurodevelopment, muscle, and detoxification proteins, but these have yet to be validated. These results provide insights into agriculture as a stressor for bumblebees, and signal for conservation action in light of ongoing anthropogenic changes.

Fecundity is arguably one of the most important life history traits, as it is closely tied to fitness. Most arthropods are recognized for their extreme reproductive capacity. For example, a single female of the oriental fruit fly Bactrocera dorsalis, a highly invasive species that is one of the most destructive agricultural pests worldwide, can lay more than 3000 eggs during its life span. The ovary is crucial for insect reproduction and its development requires further investigation at the molecular level. We report here that miR-309a is a regulator of ovarian development in B. dorsalis. Our bioinformatics and molecular studies have revealed that miR-309a binds the transcription factor pannier (GATA-binding factor A/pnr), and this activates yolk vitellogenin 2 (Vg 2) and vitellogenin receptor (VgR) advancing ovarian development. We further show that miR-309a is under the control of juvenile hormone (JH) and independent from 20-hydroxyecdysone. Thus, we identified a JH-controlled miR-309a/pnr axis that regulates Vg2 and VgR to control the ovarian development. This study has further enhanced our understanding of molecular mechanisms governing ovarian development and insect reproduction. It provides a background for identifying targets for controlling important Dipteran pests.

Polyploidy was induced in Melia volkensii (Mukau), a valuable native tree from the semi-arid regions of East Africa. Cotyledons of diploid M. volkensii (2n = 2x = 28) were treated with oryzalin for 0 (control), 1, 2, or 3 h with or without pretreatment with 1.1 µM thidiazuron. Cotyledons treated with 10 mg·L−1 oryzalin for three hours yielded 40% tetraploids. Pretreatment of cotyledons with thidiazuron for 18 days followed by treatment with oryzalin increased tetraploid plant production to 52.5%, but this also yielded more mixoploids. Compared to diploid M. volkensii, the tetraploid in vitro and young potted plants were compacter, with thicker stems, wider leaves, and a low density of longer and wider stomata. In the coming years, tetraploid M. volkensii plants will be observed in field trials and serve as a basis for further breeding efforts.

We used a population genomic approach to unravel the population structure, genetic differentiation, and genetic diversity of three widespread wild bee species across the Iberian Peninsula, Andrena agilissima, Andrena flavipes and Lasioglossum malachurum. Our results demonstrated that genetic lineages in the Ebro River valley or near the Pyrenees mountains are different from the rest of Iberia. This relatively congruent pattern across species once more supports the hypothesis of “refugia within refugia” in the Iberian Peninsula. The results for A. flavipes and A. agilissima showed an unexpected pattern of genetic differentiation, with the generalist polylectic A. flavipes having lower levels of genetic diversity (Ho = 0.0807, He = 0.2883) and higher differentiation (FST = 0.5611), while the specialist oligolectic A. agilissima had higher genetic diversity (Ho = 0.2104, He = 0.3282) and lower differentiation values (FST = 0.0957). For L. malachurum, the smallest and the only social species showed the lowest inbreeding coefficient (FIS = 0.1009) and the lowest differentiation level (FST = 0.0663). Overall, our results, suggest that this pattern of population structure and genetic diversity could be explained by the combined role of past climate changes and the life-history traits of the species (i.e., size, sociality and host-plant specialization), supporting the role of the Iberian refugia as a biodiversity hotspot.

Jonathan Willow et al. use dsαCOP-treated anthers to examine RNAi efficacy against pollen beetle, a major pest of oilseed rape. The authors found that chronic feeding resulted in significantly greater mortality compared to short-term feeding; and significant silencing of αCOP was detected only in beetles chronically exposed to dsαCOP.

Zhang et al. show that the circadian gene coupling between night feeding and day detoxification is regulated through the binding of circadian elements to E-boxes in Spodoptera litura, one of the most difficult Asian agricultural pests to control. Exposure of these larvae to a pesticide affects them more at night than during the day, suggesting the need for time-of-day considerations for pesticide application.

ABSTRACT Snodgrassella is a genus of Betaproteobacteria that lives in the gut of honeybees (Apis spp.) and bumblebees (Bombus spp). It is part of a conserved microbiome that is composed of a few core phylotypes and is essential for bee health and metabolism. Phylogenomic analyses using whole-genome sequences of 75 Snodgrassella strains from 4 species of honeybees and 14 species of bumblebees showed that these strains formed a monophyletic lineage within the Neisseriaceae family, that Snodgrassella isolates from Asian honeybees diverged early from the other species in their evolution, that isolates from honeybees and bumblebees were well separated, and that this genus consists of at least seven species. We propose to formally name two new Snodgrassella species that were isolated from bumblebees: i.e., Snodgrassella gandavensis sp. nov. and Snodgrassella communis sp. nov. Possible evolutionary scenarios for 107 species- or group-specific genes revealed very limited evidence for horizontal gene transfer. Functional analyses revealed the importance of small proteins, defense mechanisms, amino acid transport and metabolism, inorganic ion transport and metabolism and carbohydrate transport and metabolism among these 107 specific genes. IMPORTANCE The microbiome of honeybees (Apis spp.) and bumblebees (Bombus spp.) is highly conserved and represented by few phylotypes. This simplicity in taxon composition makes the bee’s microbiome an emergent model organism for the study of gut microbial communities. Since the description of the Snodgrassella genus, which was isolated from the gut of honeybees and bumblebees in 2013, a single species (i.e., Snodgrassella alvi), has been named. Here, we demonstrate that this genus is actually composed of at least seven species, two of which (Snodgrassella gandavensis sp. nov. and Snodgrassella communis sp. nov.) are formally described and named in the present publication. We also report the presence of 107 genes specific to Snodgrassella species, showing notably the importance of small proteins and defense mechanisms in this genus.

Climate, landscape composition, management practice, and wild bee pollination are all variables thought to play significant roles in commercial apple production. However, how these variables affect production efficiency under field-realistic conditions has not been investigated at large geographical scales. We combined intensive standardized field surveys (using netting and pan traps) with structural equation models to explore the relative impact of biotic and abiotic variables on bee diversity, apple yield and fruit quality, and their ability to represent reliable proxies of apple production. Here we show that apple yields are mainly driven by management practice, without evidence for a significantly superior contribution by managed honey bees. Total wild bee diversity, while negatively correlated with honey bee dominance, promoted apple quality by enhancing seed set number. Our study demonstrates that even across a broad geographical range there is potential to harness wild bee diversity as nature-based solution and as a substitute to an exclusive reliance on honey bees in the context of commercial apple production.

Abstract Wild bees are in decline on a local to global scale. The presence of managed honey bees can lead to competition for resources with wild bee species, which has not been investigated so far for human-modified landscapes. In this study we assess if managed honey bee hive density influence nest development (biomass) of bumble bees, an important trait affecting fitness. We hypothesize that domesticated honey bees can negatively affect Bombus terrestris nest development in human-modified landscapes. In Flanders, Belgium, where such landscapes are dominantly present, we selected 11 locations with landscape metrics ranging from urban to agricultural. The bee hive locations were mapped and each location contained one apiary dense (AD) and one apiary sparse (AS) study site (mean density of 7.6 ± 5.7 managed honey bee hives per km2 in AD sites). We assessed the effect of apiary density on the reproduction of reared B. terrestris nests. Reared B. terrestris nests had more biomass increase over 8 weeks in apiary sparse (AS) sites compared to nests located in apiary dense (AD) sites. This effect was mainly visible in urban locations, where nest in AS sites have 99.25 ± 60.99 g more biomass increase compared to nest in urban AD sites. Additionally, we found that managed bumble bee nests had higher biomass increase in urban locations. We conclude that the density of bee hives is a factor to consider in regard to interspecific competition between domesticated honey bees and bumble bees.

Abstract Background The western flower thrips, Frankliniella occidentalis (Pergande), is a globally invasive pest and plant virus vector on a wide array of food, fiber, and ornamental crops. The underlying genetic mechanisms of the processes governing thrips pest and vector biology, feeding behaviors, ecology, and insecticide resistance are largely unknown. To address this gap, we present the F. occidentalis draft genome assembly and official gene set. Results We report on the first genome sequence for any member of the insect order Thysanoptera. Benchmarking Universal Single-Copy Ortholog (BUSCO) assessments of the genome assembly (size = 415.8 Mb, scaffold N50 = 948.9 kb) revealed a relatively complete and well-annotated assembly in comparison to other insect genomes. The genome is unusually GC-rich (50%) compared to other insect genomes to date. The official gene set (OGS v1.0) contains 16,859 genes, of which ~ 10% were manually verified and corrected by our consortium. We focused on manual annotation, phylogenetic, and expression evidence analyses for gene sets centered on primary themes in the life histories and activities of plant-colonizing insects. Highlights include the following: (1) divergent clades and large expansions in genes associated with environmental sensing (chemosensory receptors) and detoxification (CYP4, CYP6, and CCE enzymes) of substances encountered in agricultural environments; (2) a comprehensive set of salivary gland genes supported by enriched expression; (3) apparent absence of members of the IMD innate immune defense pathway; and (4) developmental- and sex-specific expression analyses of genes associated with progression from larvae to adulthood through neometaboly, a distinct form of maturation differing from either incomplete or complete metamorphosis in the Insecta. Conclusions Analysis of the F. occidentalis genome offers insights into the polyphagous behavior of this insect pest that finds, colonizes, and survives on a widely diverse array of plants. The genomic resources presented here enable a more complete analysis of insect evolution and biology, providing a missing taxon for contemporary insect genomics-based analyses. Our study also offers a genomic benchmark for molecular and evolutionary investigations of other Thysanoptera species.

Abstract Background Long non-coding RNAs (lncRNAs) are involved in many fundamental biological processes, such as transcription regulation, protein degradation, and cell differentiation. Information on lncRNA in the melon fly, Zeugodacus cucurbitae (Coquillett) is currently limited. Results We constructed 24 RNA-seq libraries from eight tissues (midgut, Malpighian tubules, fat body, ovary, and testis) of Z. cucurbitae adults. A total of 3124 lncRNA transcripts were identified. Among those, 1464 were lincRNAs, 1037 were intronic lncRNAs, 301 were anti-sense lncRNAs, and 322 were sense lncRNAs. The majority of lncRNAs contained two exons and one isoform. Differentially expressed lncRNAs were analyzed between tissues, and Malpighian tubules versus testis had the largest number. Some lncRNAs exhibited strong tissue specificity. Specifically expressed lncRNAs were identified and filtered in tissues of female and male Z. cucurbitae based on their expression levels. Four midgut-specific lncRNAs were validated by quantitative real-time polymerase chain reaction (RT-qPCR), and the data were consistent with RNA-seq data. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of targets of midgut-specific lncRNAs indicated an enrichment of the metabolic process. Conclusions This was the first systematic identification of lncRNA in the melon fly. Expressions of lncRNAs in multiple adult tissues were evaluated by quantitative transcriptomic analysis. These qualitative and quantitative analyses of lncRNAs, especially the tissue-specific lncRNAs in Z. cucurbitae, provide useful data for further functional studies.

Habitat loss and fragmentation, and the effects of pesticides, contribute to biodiversity losses and unsustainable food production. Given the United Nation’s (UN’s) declaration of this decade as the UN Decade on Ecosystem Restoration, we advocate combining conservation biocontrol-enhancing practices with the use of RNA interference (RNAi) pesticide technology, the latter demonstrating remarkable target-specificity via double-stranded (ds)RNA’s sequence-specific mode of action. This specificity makes dsRNA a biosafe candidate for integration into the global conservation initiative. Our interdisciplinary perspective conforms to the UN’s declaration, and is facilitated by the Earth BioGenome Project, an effort valuable to RNAi development given its utility in providing whole-genome sequences, allowing identification of genetic targets in crop pests, and potentially relevant sequences in non-target organisms. Interdisciplinary studies bringing together biocontrol-enhancing techniques and RNAi are needed, and should be examined for various crop‒pest systems to address this global problem.

Concerns regarding the ecological and health risks posed by synthetic insecticides have instigated the exploration of alternative methods for controlling insects, such as entomopathogenic fungi (EPF) as biocontrol agents. Therefore, this review discusses their use as a potential alternative to chemical insecticides and especially focuses on the two major ones, Beauveria bassiana and Metarhizium anisopliae, as examples. First, this review exemplifies how B. bassiana- and M. anisopliae-based biopesticides are used in the world. Then, we discuss the mechanism of action by which EPF interacts with insects, focusing on the penetration of the cuticle and the subsequent death of the host. The interactions between EPF and the insect microbiome, as well as the enhancement of the insect immune response, are also summarized. Finally, this review presents recent research that N-glycans may play a role in eliciting an immune response in insects, resulting in the increased expression of immune-related genes and smaller peritrophic matrix pores, reducing insect midgut permeability. Overall, this paper provides an overview of the EPF in insect control and highlights the latest developments relating to the interaction between fungi and insect immunity.

The unprecedented target-specificity of double-stranded RNA (dsRNA), due to its sequence-specific mode of action, puts dsRNA at the forefront of biosafe insecticide technology. Since 2007, sensitive target genes have been identified in numerous crop pest insects, with an end goal of applying RNA interference (RNAi) in pest management. Key RNAi targets identified include genes involved in (1) feeding and digestion, (2) production of dsRNases, (3) resistance to insecticides and plant allelochemicals, (4) reproductive fitness, and (5) transmission of plant viruses. Despite the advances, there remain critical knowledge gaps in each of these areas. Particular emphasis must be placed on ensuring RNAi's compatibility with integrated pest management (IPM), via further identification of molecular targets that reduce crop damage while sustaining pest (host) populations for highly specialized biocontrol agents, the latter representing a core pillar of IPM.

Forest insects are emerging in large extension in response to ongoing climatic changes, penetrating geographic barriers, utilizing novel hosts, and influencing many hectares of conifer forests worldwide. Current management strategies have been unable to keep pace with forest insect population outbreaks, and therefore novel and aggressive management strategies are urgently required to manage forest insects. RNA interference (RNAi), a Noble Prize-winning discovery, is an emerging approach that can be used for forest protection. The RNAi pathway is triggered by dsRNA molecules, which, in turn, silences genes and disrupts protein function, ultimately causing the death of the targeted insect. RNAi is very effective against pest insects; however, its proficiency varies significantly among insect species, tissues, and genes. The coleopteran forest insects are susceptible to RNAi and can be the initial target, but we lack practical means of delivery, particularly in systems with long-lived, endophagous insects such as the Emerald ash borer, Asian longhorn beetles, and bark beetles. The widespread use of RNAi in forest pest management has major challenges, including its efficiency, target gene selection, dsRNA design, lack of reliable dsRNA delivery methods, non-target and off-target effects, and potential resistance development in wood-boring pest populations. This review focuses on recent innovations in RNAi delivery that can be deployed against forest pests, such as cationic liposome-assisted (lipids), nanoparticle-enabled (polymers or peptides), symbiont-mediated (fungi, bacteria, and viruses), and plant-mediated deliveries (trunk injection, root absorption). Our findings guide future risk analysis of dsRNA-based forest protection products (FPPs) and risk assessment frameworks incorporating sequence complementarity-based analysis for off-target predictions. This review also points out barriers to further developing RNAi for forest pest management and suggests future directions of research that will build the future use of RNAi against wood-boring coleopterans.

This article is part of the Special Issue Glycosylation—The Most Diverse Post-Translational Modification [...]

The biosafety aspect of applying double-stranded RNA (dsRNA) in crop pest management is rooted in dsRNA's mode of action, which displays nucleotide sequence-specificity to a particular region of a messenger RNA (mRNA), against which the insecticidal dsRNA is designed. This prominent and promising class of insecticides therefore has the potential to target a single pest species while conferring negligible effect on non-target organisms. Recent studies examining the effect of target-specific dsRNA in adults of the pollen beetle Brassicogethes aeneus, a major pest of oilseed rape (Brassica napus) crops in Europe, suggest the potential for developing a gene-silencing approach within integrated B. aeneus management. The present study examines the efficacy of target-specific dsRNA on target-mRNA silencing, and subsequent gene-silencing-induced mortality, in B. aeneus larvae, as this life stage represents a critical target for achieving optimal integrated B. aeneus control. Treatment applications occurred via feeding on dsRNA-treated anthers for 3 d. We observed variable gene-silencing efficacy, all target treatments having a significant or marginally significant effect after 3 d of dsRNA feeding, with greater variability at 6 d. These results further validated significant gene-silencing-induced mortality observed for one of the target treatments. Moreover, gene-silencing-induced mortality occurred at a quicker rate in B. aeneus larvae compared to what has been previously observed in B. aeneus adults. Finally, we consider refinements that must be made to B. aeneus larval bioassay setups to promote and strengthen future larval studies regarding this important crop pest species.

Risk assessment of pesticides involves ecotoxicological testing. In case pesticide exposure to bees is likely, toxicity tests are performed with honey bees (Apis mellifera), with a tiered approach, for which validated and internationally accepted test protocols exist. However, concerns have grown regarding the protection of non-Apis bees [bumble bees (Bombus spp.), solitary and stingless bees], given their different life cycles and therefore distinct exposure routes. Larvae of solitary bees of the genus Osmia feed on unprocessed pollen during development, yet no toxicity test protocol is internationally accepted or validated to assess the impact of pesticide exposure during this stage of their life cycle. Therefore, the purpose of this study is to further validate a test protocol with two solitary bee species (O. cornuta and O. bicornis) to assess lethal and sublethal effects of pesticide exposure on larval development. Larvae were exposed to thiacloprid (neonicotinoid insecticide) mixed in a new, artificial pollen provision. Both lethal (developmental and winter mortality) and sublethal endpoints (larval development time, pollen provision consumption, cocoon weight, emergence time and adult longevity) were recorded. Effects of lower, more environmentally realistic doses were only reflected in sublethal endpoints. In both bee species, thiacloprid treatment was associated with increased developmental mortality and larval development time, and decreased pollen provision consumption and cocoon weight. The test protocol proved valid and robust and showed that for higher doses of thiacloprid the acute endpoint (larval mortality) is sufficient. In addition, new insights needed to develop a standardized test protocol were acquired, such as testing of a positive control for the first time and selection of male and female individuals at egg level.

Protein glycosylation is one of the most common and most important post-translational modifications. Despite the growing knowledge on N-glycosylation, the research on O-glycosylation is lagging behind. This study investigates the importance of O-glycosylation in the post-embryonic development of insects using the red flour beetle, Tribolium castaneum, as a model. We identified 28 O-glycosylation-related genes (OGRGs) in the genome of the red flour beetle. 14 OGRGs were selected for functional analysis based on their involvement in the initial attachment of the carbohydrate in the different O-glycosylation pathways or the further elongation of the most abundant O-glycans and, in addition, showing severe RNAi-induced phenotypes in Drosophila melanogaster. The expression profile of these OGRGs was mapped throughout the developmental stages of the insect and in the different tissues of the pupa and adult. Subsequently, these genes were silenced using RNA interference (RNAi) to analyze their role in development. A broad spectrum of phenotypes was observed: from subtle effects and disrupted wing formation when silencing the genes involved in O-mannosylation, to blockage of pupation and high mortality after silencing of the genes involved in O-GalNAc and core 1 O-glycan (O-GalNAc-Gal) synthesis. RNAi experiments were also performed to assess the effects of blocking multiple pathways of O-glycosylation. However, the observed phenotypes induced by multiple RNAi were similar to those of the single gene RNAi experiments. The silencing of OGRGs often resulted in high mortality and wing phenotypes, indicating the importance of O-glycosylation for the survival of the insect and the formation of wings during the post-embryonic development of T. castaneum.