Phytochemical screening and insecticidal activities of some medicinal plants against the maize weevil, Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae).

This study emphasizes the phytochemical study of some locally available botanicals against maize weevils. Nine plant parts were collected from six plant species. The test plant powder (200 g) was suspended sequentially in 600 ml of petroleum ether, chloroform, acetone, methanol, and distilled water for 72 h with frequent agitation. Different concentrations of the crude extracts were applied to maize seeds at rates of 10 ml, 15 ml and 20 ml per 100 g. All treatments with different extracts at different rates of application showed significant differences (p < 0.05) in the cumulative mean percentage mortality of the maize weevil. The seed extract of Maesa lanceolata and Croton macrostachyus and the leaf extract of Clausena anisata showed cumulative percent mortality ranged 95.32-98.02% in 28 days after treatment application. There was no significant difference (p > 0.05) among all treatments for the prevention of F1 progeny emergence. In all extracts, Clausena anisata showed 100% inhibition of F1 progeny emergence. All treatments significantly reduced seed weight loss and damage. The treated maize seeds were germinated with an acceptable germination quality. In conclusion, an increased dosage of the extract resulted in significant mortality in maize weevils. The seed extracts of Maesa lanceolata and Croton macrostachyus and Clausena anisata leaf extract were observed to be the most promising botanical in protecting stored maize against maize weevil.

Evaluation of heterologous expression in Pichia pastoris of Pine Weevil TRPA1 by GFP and flow cytometry.

BACKGROUND: The wasabi receptor, also known as the Transient Receptor Potential Ankyrin 1 (TRPA1) ion channel, is a potential target for development of repellents for insects, like the pine weevil (Hylobius abietis) feeding on conifer seedlings and causing damage in forestry. Heterologous expression of TRPA1 from pine weevil in the yeast Pichia pastoris can potentially provide protein for structural and functional studies. Here we take advantage of the Green Fluorescent Protein (GFP) tag to examine the various steps of heterologous expression, to get more insight in clone selection, expression and isolation of the intact purified protein. RESULTS: The sequence of HaTRPA1 is reported and GFP-tagged constructs were made of the full-length protein and a truncated version (Δ1-708 HaTRPA1), lacking the N-terminal ankyrin repeat domain. Clones were screened on GFP expression plates, induced in small liquid cultures and in fed-batch cultures, and evaluated by flow cytometry and fluorescence microscopy. The screening on plates successfully identifies low-expression clones, but fails to predict the ranking of the best performing clones in small-scale liquid cultures. The two constructs differ in their cellular localization. Δ1-708 HaTRPA1 is found in a ring at the perimeter of cell, whereas HaTRPA1 is forming highly fluorescent speckles in interior regions of the cell. The pattern is consistent in different clones of the same construct and persists in fed-batch culture. The expression of Δ1-708 HaTRPA1 decreases the viability more than HaTRPA1, and in fed-batch culture it is clear that intact cells first express Δ1-708 HaTRPA1 and then become damaged. Purifications show that both constructs suffer from degradation of the expressed protein, but especially the HaTRPA1 construct. CONCLUSIONS: The GFP tag makes it possible to follow expression by flow cytometry and fluorescence microscopy. Analyses of localization, cell viability and expression show that the former two parameters are specific for each of the two evaluated constructs, whereas the relative expression of the constructs varies with the cultivation method. High expression is not all that matters, so taking damaged cells into account, something that may be linked to protein degradation, is important when picking the most suitable construct, clone, and expression scheme.

Seeds of nonhost species as sources of toxic compounds for the cowpea weevil Callosobruchus maculatus (F.).

Cowpea weevil, Callosobruchus maculatus, is the primary pest of stored cowpea seeds. The management of this infestation currently relies on insecticides, resulting in environmental pollution and selection of insecticide-resistant pests. Consequently, research efforts are being devoted to identify natural insecticides as sustainable and environment friendly alternatives for the control of C. maculatus. In this study, we explore the toxic effects of the nonhost seeds Parkia multijuga, Copaifera langsdorffii, Ormosia arborea, Amburana cearensis, Lonchocarpus guilleminianus, Sapindus saponaria, and Myroxylon peruiferum, on the cowpea weevil C. maculatus. Notably, all nonhost seeds led to reductions between 60 and 100% in oviposition by C. maculatus females. Additionally, the larvae were unable to penetrate the nonhost seeds. Artificial seeds containing 0.05% to 10% of cotyledon flour were toxic to C. maculatus larvae. Approximately 40% of larvae that consumed seeds containing 0.05% of O. arborea failed to develop, in contrast to control larvae. Proteomic analysis of A. cearensis and O. arborea seeds identify revealed a total of 371 proteins. From those, 237 are present in both seeds, 91 were exclusive to O. arborea seeds, and 43 were specific to A. cearensis seeds. Some of these proteins are related to defense, such as proteins containing the cupin domain and 11S seed storage protein. The in silico docking of cupin domain-containing proteins and 11S storage protein with N-acetylglucosamine (NAG)4 showed negative values of affinity energy, indicating spontaneous binding. These results showed that nonhost seeds have natural insecticide compounds with potential to control C. maculatus infestation.

Potential of Thuja occidentalis L. Essential Oil and Water Extracts against Field Crop Pests.

Thuja occidentalis L. essential oil (EOTO) and its compounds, such as terpinyl acetate, bornyl acetate, and ß-thujone, are claimed to be highly effective against some storage pests, sanitary insects, or pests of fruit trees, while data about its use in protecting field crops are very scarce. There is also a lack of information in the literature about the insecticidal value of water extracts from T. occidentalis (WETOs). Both essential oils (EOs) and water extracts (WEs) from various plants have advantages and disadvantages in terms of their use as insecticides. EOs are generally more effective, but their preparation is more complicated and quite expensive. In turn, WEs are simple to prepare and cheap, but they often have limited effectiveness. Moreover, significant differences in responses exist depending on the species of the donor plant, the method of preparing the extract, its concentration, the species of the pest being controlled, the developmental stage, and even the gender of the pest. The goals of the research were to assess the effect of EOTO and WETOs prepared from dry and fresh matter on the mortality, feeding, and body mass changes of important crop pests, i.e., the black bean aphid, pea leaf weevil, and Colorado potato beetle (CPB), respectively, as well as on the mortality and voracity of non-target organism Asian lady beetle young larvae. EOTO showed significant aphicidal activity with LC50 = 0.8267% and 0.2453% after 42 h of the experiment for nymphs and wingless females of black bean aphid, respectively. Adults of CPB were more resistant to EOTO than aphids, with LC50 values for females equal to 1.5327% and 1.3113% after 48 h and after 72 h of the experiment. There was no significant effect of EOTO on CPB foraging. Calculated LC50 values for pea leaf weevil adults were lower than those for CPB (0.9638% and 0.8573% for males after 12 h and 24 h, respectively). In the case of this pest, a clear reduction in foraging was obtained, with higher concentrations of EOTO resulting in more pronounced reductions in foraging behavior. Concentrations of EOTO above 0.5%, which showed efficacy against the aphid, were lethal to 3-day-old larvae of the Asian lady beetle. WETOs, in turn, showed significant potential in inhibiting adult pea leaf weevil feeding, with very low or no effectiveness in reducing A. fabae and CPB, respectively.

Assessment and Optimization of the Insecticidal Properties of γ-Al2O3 Nanoparticles Derived from Mentha pulegium By-Products to Xylosandrus crassiusculus (Carob Beetle).

This study concentrates on assessing the insecticidal attributes of the γ-Al2O3 nanoparticles derived from the remnants of Mentha pulegium, which include essential oil, ethanolic extract, and plant waste. The synthesis of the γ-Al2O3 nanoparticles was executed using a direct sol-gel procedure, affirming the crystal structure according to extensive physicochemical analyses such as UV-Vis, XRD, FTIR, and SEM. Evaluation of the insecticidal activity in vitro was conducted against Xylosandrus crassiusculus, a pest that infests carob wood, utilizing strains from diverse forests in the Khenifra region, situated in the Moroccan Middle Atlas. The lethal doses 50 ranged from 40 mg/g to 68 mg/g, indicating moderate effectiveness compared to the commercial insecticide Permethrin. Optimization of the conditions for the efficiency of the γ-Al2O3 nanoparticles was determined using experimental plans, revealing that time, humidity, and temperature were influential factors in the lethal dose 50 of these nanomaterials. Moreover, this study encompasses the establishment of correlations using Principal Component Analysis (PCA) and Ascending Hierarchical Classification (AHC) among various geographic, biological, and physical data, amalgamating geographic altitude and γ-Al2O3 nanoparticle insecticide parameters, as well as the attributes of the mechanical tests conducted on the carob wood affected by insects. The correlations highlight the close connections between the effectiveness of the insecticide, mountain altitude, and the mechanical parameters that were examined. Ultimately, these nanoparticles demonstrate promising potential as alternative insecticides, thus opening up encouraging prospects for safeguarding against carob wood pests.

Predicting the dispersal and invasion dynamics of ambrosia beetles through demographic reconstruction and process-explicit modeling.

Evaluating potential routes of invasion of pathogens and vectors of sanitary importance is essential for planning and decision-making at multiple scales. An effective tool are process-explicit models that allow coupling environmental, demographic and dispersal information to evaluate population growth and range dynamics as a function of the abiotic conditions in a region. In this work we simulate multiple dispersal/invasion routes in Mexico that could be taken by ambrosia beetles and a specific symbiont, Harringtonia lauricola, responsible for a severe epiphytic of Lauraceae in North America. We used Xyleborus bispinatus Eichhoff 1868 as a study subject and estimated its demography in the laboratory in a temperature gradient (17, 20, 26, 29, 35 °C), which we then used to parameterize a process-based model to estimate its metapopulation dynamics. The maximum intrinsic growth rate of X. bispinatus is 0.13 with a thermal optimum of 26.2 °C. The models suggest important regions for the establishment and dispersal the states of Veracruz, Chiapas and Oaxaca (high host and secondary vectors diversity), the Isthmus of Tehuantepec (connectivity region), and Michoacán and Jalisco (important avocado plantations). The use of hybrid process-based models is a promising tool to refine the predictions applied to the study of biological invasions and species distributions.

Drought increases Norway spruce susceptibility to the Eurasian spruce bark beetle and its associated fungi.

Drought affects the complex interactions between Norway spruce, the bark beetle Ips typographus and associated microorganisms. We investigated the interplay of tree water status, defense and carbohydrate reserves with the incidence of bark beetle attack and infection of associated fungi in mature spruce trees. We installed roofs to induce a 2-yr moderate drought in a managed spruce stand to examine a maximum of 10 roof and 10 control trees for resin flow (RF), predawn twig water potentials, terpene, phenolic and carbohydrate bark concentrations, and bark beetle borings in field bioassays before and after inoculation with Endoconidiophora polonica and Grosmannia penicillata. Drought-stressed trees showed more attacks and significantly longer fungal lesions than controls, but maintained terpene resin defenses at predrought levels. Reduced RF and lower mono- and diterpene, but not phenolic concentrations were linked with increased host selection. Bark beetle attack and fungi stimulated chemical defenses, yet G. penicillata reduced phenolic and carbohydrate contents. Chemical defenses did not decrease under mild, prolonged drought in our simulated small-scale biotic infestations. However, during natural mass attacks, reductions in carbon fixation under drought, in combination with fungal consumption of carbohydrates, may deplete tree defenses and facilitate colonization by I. typographus.

Diel flight activity in response to attractants in the predatory beetle, Cathartus quadricollis (Coleoptera: Silvanidae) in macadamia nut in Hawaii.

The diel flight activity in Cathartus quadricollis (Guerin-Meneville) (Coleoptera: Silvanidae), a predator of two important pests in Hawaii, coffee berry borer, Hypothenemus hampei (Ferrari) and tropical nut borer, Hypothenemus obscurus (F.) (Coleoptera: Curculionidae: Scolytinae) was studied in a macadamia nut orchard using yellow sticky traps baited with pheromone and fungal volatile attractants. The study was conducted at different months throughout the year and at different times during the lunar cycle (new moon and full moon). Flight activity peaked in the late hours of the photophase into the early hours of the scotophase, between 1830 and 2000 h; flight activity also occurred but to a lesser extent in the early morning hours between 0700 and 1030 h. Numbers of captured C. quadricollis during periods of flight activity were negatively correlated with wind speed. The implications of these findings for the development of optimal pest management strategies including biological control are discussed.

First record of non-native Xylosandrus compactus and Anisandrus maiche (Coleoptera: Curculionidae, Scolytinae) in Slovenia.

The introduction of non-native organisms into new areas may pose major threats to natural ecosystems. Therefore, continuous and careful national surveys for the interception of invasive non-native species are necessary. During the national survey of quarantine species in 2023, two new non-native ambrosia beetles, Anisandrus maiche (Kurentzov) and Xylosandrus compactus (Eichhoff), were recorded in Slovenia. Anisandrus maiche was recorded in three locations in the eastern part of Slovenia, with a total of 386 individuals. Three individuals of Xylosandrus compactus were collected in one location near the port of Koper. We discuss the possible pathways of introduction and dispersal.

Taxonomic changes for Neotropical pinhole borer ambrosia beetles (Coleoptera: Curculionidae: Platypodinae).

Stephen L. Wood re-defined Platypus such that its members are native to realms outside of the Americas and transferred most Neotropical species out of that genus. I have come across 44 species that still remain, though, and these are treated here. In total, I report 49 new generic assignments, 30 of which are transfers out of Platypus. I propose 22 new synonymies, eight of which are Platypus species that are synonymized with previously transferred species. Six Neotropical species are left in Platypus, for reasons detailed in the text. These taxonomic acts affect the compositions of eight of the 11 Neotropical genera of core Platypodinae. The following species are transferred from Platypus Herbst, 1793: Cenocephalus dubiosus (Schedl, 1933) comb. nov., Cenocephalus neotruncatus (Schedl, 1972) comb. nov.; Costaroplatus barbosai (Schedl, 1972) comb. nov., Costaroplatus devius (Schedl, 1976) comb. nov., Costaroplatus mixtus (Schedl, 1976) comb. nov., Costaroplatus roppai (Schedl, 1978) comb. nov.; Epiplatypus bicaudatulus (Schedl, 1935) comb. nov., Epiplatypus carduus (Schedl, 1936) comb. nov., Epiplatypus complanus (Schedl, 1967) comb. nov., Epiplatypus grandiporus (Schedl, 1961) comb. nov., Epiplatypus insculptus (Schedl, 1967) comb. nov., Epiplatypus macroporus (Chapuis, 1865) comb. nov., Epiplatypus perforans (Schedl, 1961) comb. nov., Epiplatypus propinquus (Schedl, 1959) comb. nov., Epiplatypus quadrispinatus (Chapuis, 1865) comb. nov., Epiplatypus sallei (Chapuis, 1865) comb. nov., Epiplatypus sequius (Schedl, 1935) comb. nov.; Euplatypus detectus (Schedl, 1976) comb. nov., Euplatypus erraticus (Schedl, 1972) comb. nov., Euplatypus longulus (Chapuis, 1865) comb. nov., Euplatypus perplexus Bright, 1972 comb. nov., Euplatypus rugosifrons (Schedl, 1933) comb. nov., Euplatypus vexans (Schedl, 1972) comb. nov.; Megaplatypus asperatus (Schedl, 1976) comb. nov., Megaplatypus carinifer (Schedl, 1970), Megaplatypus durus (Schedl, 1936) comb. nov., Megaplatypus eversus (Wood, 1971) comb. nov., Megaplatypus gagates (Schedl, 1976) comb. nov., Megaplatypus irrepertus (Schedl, 1936) comb. nov., Megaplatypus lineaticornis (Schedl, 1936) comb. nov., Megaplatypus paramonovi (Schedl, 1972) comb. nov., Megaplatypus schedli (Wood, 1966) comb. nov., Megaplatypus vafer (Schedl, 1972) comb. nov.; Teloplatypus caligatus (Schedl, 1972) comb. nov. Costaroplatus bidens (Schedl, 1970) comb. nov. and Costaroplatus darlingtoni (Reichardt, 1965) comb. nov. are transferred from Megaplatypus Wood, 1993. Costaroplatus vonfaberi (Reichardt, 1962) comb. nov. is transferred from Platyphysus Wood, 1993. Epiplatypus striatus (Chapuis, 1865) comb. nov., Megaplatypus contextus (Schedl, 1963) comb. nov., Megaplatypus decorus (Schedl, 1936) comb. nov. and Megaplatypus dignatus (Schedl, 1936) comb. nov. are removed from Euplatypus Wood, 1993. Epiplatypus ornatus (Schedl, 1936) comb. nov. is transferred from Teloplatypus Wood, 1993. Euplatypus jamaicensis Bright, 1972 comb. nov., Megaplatypus discolor (Blandford, 1896) comb. nov., Teloplatypus brasiliensis (Nunberg, 1959) comb. nov., Teloplatypus nudus (Schedl, 1936) comb. nov. and Teloplatypus pernudus (Schedl, 1936) comb. nov. are transferred from Epiplatypus Wood, 1993. Costaroplatus ornatus (Schedl, 1936) comb. nov., is transferred from Cenocephalus Chapuis, 1865. Megaplatypus acutidens (Blandford, 1895) comb. nov. and Megaplatypus despectus (Schedl, 1971) comb. nov. are transferred from Tesserocerus Saunders, 1837. New synonymies are proposed as follows: Cenocephalus rugicollis Schedl, 1952 (= Cenocephalus epistomalis Wood, 1966 syn. nov.); Tesserocerus forcipatus Schedl, 1972 (= Platypus aplanatus Schedl, 1976 syn. nov.); Tesserocerus retusus Gurin-Mneville, 1838 (= Tesserocerus guerini ssp. montanus Schedl, 1960 syn. nov.); Tesserocerus simulatus Schedl, 1936 (= Platypus bilobus Schedl, 1961 syn. nov.); Tesserocerus spinax Blandford, 1896 (= Tesserocephalus forficula Schedl, 1936 syn. nov.); Costaroplatus carinulatus (Chapuis, 1865) (= Platypus umbrosus Schedl, 1936 syn. nov.); Costaroplatus shenefelti Nunberg (1963) (= Platypus abditulus Wood, 1966 syn. nov.); Costaroplatus vonfaberi (Reichardt, 1962) (= Platypus convexus Schedl, 1972 syn. nov.); Epiplatypus sallei (Chapuis, 1865) (= Platypus quadricaudatulus Schedl, 1934 syn. nov. and = Platypus filaris Wood, 1971 syn. nov.); Euplatypus longulus (Chapuis, 1865) (= Platypus dimidiatus Chapuis, 1865 syn. nov. = Platypus mulsanti Chapuis, 1865 syn. nov. and = Platypus pseudolongulus Schedl, 1963 syn. nov. ); Megaplatypus acutidens (Blandford, 1895) (= Tesserocerus alternantes Schedl, 1977 syn. nov.); Megaplatypus durus (Schedl, 1936) (= Platypus arcuatus Schedl, 1976 syn. nov.); Megaplatypus fuscus (Chapuis, 1865) (= Platypus marginatus Chapuis, 1865 syn. nov., = Platypus granarius Schedl, 1952 syn. nov., and = Platypus obsitus Schedl, 1976 syn. nov.); Megaplatypus irrepertus (Schedl, 1936) (= Platypus sulcipennis Schedl, 1976 syn. nov.); Neotrachyostus abbreviatus (Chapuis, 1865) (= Platypus concavus Chapuis, 1865 syn. nov.); Teloplatypus enixus (Schedl, 1936) (= Platypus interponens Schedl, 1978 syn. nov.); Teloplatypus ratzeburgi (Chapuis, 1865) (= Platypus pallidipennis Blandford, 1896 syn. nov.). Platypus simpliciformis Wood, 1966 had been transferred by Wood (1993) to both Megaplatypus and Euplatypus by mistake; I propose keeping it in Megaplatypus. Six Neotropical species are left in the genus Platypus with the status incertae sedis: Platypus armatus Chapuis, 1865; Platypus dorsalis Schedl, 1972; Playpus quadrilobus Blandford, 1895; Platypus squamifer Schedl, 1963; Platypus subaequalispinosus Schedl, 1936; and Platypus trispinosus Chapuis, 1965. These taxonomic changes prepare the foundations for future revisionary work on the American Platypodinae.

New species, new synonymy, taxonomic notes and new records of bark and ambrosia beetles from the southwestern United States and northern Mexico (Coleoptera: Curculionidae: Platypodinae and Scolytinae).

Chaetophloeus flourensiae new species, is described from the Chihuahuan Desert from Arizona and western Texas and Hylocurus incognitus new species is described from Texas, Oklahoma and Louisiana. New synonymies include: Chramesus mimosae Blackman, 1938 (= Chramesus varius Wood, 1969); Hylocurus rudis (LeConte, 1876) (= Hylocurus binodatus Wood, 1974; Hypothenemus seriatus (Eichhoff, 1872) (= Stephanoderes multidentatus Hopkins, 1915, Stephanoderes nitidifrons Hopkins, 1915, Hypothenemus hopkinsi Browne, 1963); Hypothenemus pubescens Hopkins, 1915 (= Hypothenemus sparsus Hopkins, 1915, Hypothenenus similis Hopkins, 1915, Stephanoderes tridentatus Hopkins, 1915); Phloeotribus scabricollis Hopkins, 1916 (=Phloeotribus pseudoscabricollis Atkinson, 1989; Pseudothysanoes yuccae (Wood, 1956), (=Pseudothysanoes yuccavorus Wood, 1971); and Thysanoes texanus Blackman, 1943 (=Thysanoes mexicanus Wood, 1956). Hylocurus schwarzi Blackman, 1928, is redescribed including the first description of the female. New locality and host records that significantly extend the respective ranges are included for 30 species from the border region of the United States and Mexico.

Cubanthonomus, a new genus of Anthonomini (Coleoptera: Curculionidae: Curculioninae) from Cuba.

Cubanthonomus Anderson, new genus (Coleoptera: Curculionidae: Curculioninae: Anthonomini) is here described to accommodate the Cuban species Cleogonus grossulus Suffrian, which was previously placed as incertae sedis within Cleogonini. The genus is characterized by head constricted behind the eyes, globose form, compact antennal funicle of seven antennomeres, base of pronotum strongly produced posteriorly at middle, elytra with rows of punctures (striae effaced), tarsal claws with small basal tooth, profemora with two teeth, meso- and metafemora with single small tooth. Specimens have been collected in tropical semideciduous forest but no host associations are known.

A new widely distributed invasive alien species of Amasa ambrosia beetles (Coleoptera: Curculionidae: Scolytinae: Xyleborini).

Amasa parviseta Knek & Smith, new species is described from Australia, Brazil, Uruguay, France and Spain. The species is native to Australia and appears to have spread widely in association with introduced Eucalyptus species.

Cereal weevils' antimicrobial peptides: at the crosstalk between development, endosymbiosis and immune response.

Interactions between animals and microbes are ubiquitous in nature and strongly impact animal physiology. These interactions are shaped by the host immune system, which responds to infections and contributes to tailor the associations with beneficial microorganisms. In many insects, beneficial symbiotic associations not only include gut commensals, but also intracellular bacteria, or endosymbionts. Endosymbionts are housed within specialized host cells, the bacteriocytes, and are transmitted vertically across host generations. Host-endosymbiont co-evolution shapes the endosymbiont genome and host immune system, which not only fights against microbial intruders, but also ensures the preservation of endosymbionts and the control of their load and location. The cereal weevil Sitophilus spp. is a remarkable model in which to study the evolutionary adaptation of the immune system to endosymbiosis owing to its binary association with a unique, relatively recently acquired nutritional endosymbiont, Sodalis pierantonius. This Gram-negative bacterium has not experienced the genome size shrinkage observed in long-term endosymbioses and has retained immunogenicity. We focus here on the sixteen antimicrobial peptides (AMPs) identified in the Sitophilus oryzae genome and their expression patterns in different tissues, along host development or upon immune challenges, to address their potential functions in the defensive response and endosymbiosis homeostasis along the insect life cycle. This article is part of the theme issue 'Sculpting the microbiome: how host factors determine and respond to microbial colonization'.

Immune system modulation & virus transmission during parasitism identified by multi-species transcriptomics of a declining insect biocontrol system.

BACKGROUND: The Argentine stem weevil (ASW, Listronotus bonariensis) is a significant pasture pest in Aotearoa New Zealand, primarily controlled by the parasitoid biocontrol agent Microctonus hyperodae. Despite providing effective control of ASW soon after release, M. hyperodae parasitism rates have since declined significantly, with ASW hypothesised to have evolved resistance to its biocontrol agent. While the parasitism arsenal of M. hyperodae has previously been investigated, revealing many venom components and an exogenous novel DNA virus Microctonus hyperodae filamentous virus (MhFV), the effects of said arsenal on gene expression in ASW during parasitism have not been examined. In this study, we performed a multi-species transcriptomic analysis to investigate the biology of ASW parasitism by M. hyperodae, as well as the decline in efficacy of this biocontrol system. RESULTS: The transcriptomic response of ASW to parasitism by M. hyperodae involves modulation of the weevil's innate immune system, flight muscle components, and lipid and glucose metabolism. The multispecies approach also revealed continued expression of venom components in parasitised ASW, as well as the transmission of MhFV to weevils during parasitism and some interrupted parasitism attempts. Transcriptomics did not detect a clear indication of parasitoid avoidance or other mechanisms to explain biocontrol decline. CONCLUSIONS: This study has expanded our understanding of interactions between M. hyperodae and ASW in a biocontrol system of critical importance to Aotearoa-New Zealand's agricultural economy. Transmission of MhFV to ASW during successful and interrupted parasitism attempts may link to a premature mortality phenomenon in ASW, hypothesised to be a result of a toxin-antitoxin system. Further research into MhFV and its potential role in ASW premature mortality is required to explore whether manipulation of this viral infection has the potential to increase biocontrol efficacy in future.

Genetic characterization of 2 Ceutorhynchus (Coleoptera: Curculionidae) weevils with mitogenomes and insights into the phylogeny and evolution of related weevils.

The rape stem weevil (Ceutorhynchus asper Roel.) and its close relatives primarily breed on cruciferous plants and cause severe damage to rapeseed production. However, their genetic and molecular information is still scarce. Here, we generated mitogenomes for both C. asper and Ceutorhynchus albosuturalis. The lengths of the 2 mitochondrial genomes are 14,207 bp (C. asper) and 15,373 bp (C. albosuturalis), and both weevils exhibit identical numbers of protein-coding genes with the absence of trnI. A + T contents for both mitogenomes are high (80% and 79.9%, respectively). Haplotype and genetic distance analyses showed that the genetic differentiation of C. asper populations in northwestern China is low. Based on 5 datasets from mitogenomes, phylogenetic analyses with maximum-likelihood and Bayesian methods show that both species (C. asper and C. albosuturalis) fall in the CCCMS clade (Curculioninae, Conoderinae, Cossoninae, Molytinae, and Scolytinae) of Curculionidae and belong to clades H and I of the genus Ceutorhynchus, respectively. Larvae of the clade H weevils mainly are borers in petioles or stems of cruciferous plants, while larvae of the clade I weevils mainly inhabit the fruits of the same plants, suggesting that ecological niche specialization can play a critical role in the diversification of Ceutorhynchus species. This study generates baseline molecular and genetic information for future research of Ceutorhynchus-related taxa and provides insights into the phylogeny and evolution of Curculionidae.

Evolution, types, and distribution of flight control devices on wings and elytra in bark beetles.

Gaining the ability to fly actively was a ground-breaking moment in insect evolution, providing an unprecedented advantage over other arthropods. Nevertheless, active flight was a costly innovation, requiring the development of wings and flight muscles, the provision of sufficient energetic resources, and a complex flight control system. Although wings, flight muscles, and the energetic budget of insects have been intensively studied in the last decades, almost nothing is known regarding the flight-control devices of many crucial insect groups, especially beetles (Coleoptera). Here, we conducted a phylogenetic-informed analysis of flight-related mechanosensors in 28 species of bark beetles (Curculionidae: Scolytinae, Platypodinae), an economically and ecologically important group of insects characterized by striking differences in dispersal abilities. The results indicated that beetle flight apparatus is equipped with different functional types of mechanosensors, including strain- and flow-encoding sensilla. We found a strong effect of allometry on the number of mechanosensors, while no effect of relative wing size (a proxy of flight investment) was identified. Our study constitutes the first step to understanding the drivers and constraints of the evolution of flight-control devices in Coleoptera, including bark beetles. More research, including a quantitative neuroanatomical analysis of beetle wings, should be conducted in the future.

Dietary potential of the symbiotic fungus Penicillium herquei for the larvae of a nonsocial fungus-cultivating weevil Euops chinensis.

Many insect taxa cultivate fungi for food. Compared to well-known fungus cultivation in social insects, our knowledge on fungus cultivation in nonsocial insects is still limited. Here, we studied the nutritional potentials of the fungal cultivar, Penicillium herquei, for the larvae of its nonsocial insect farmer, Euops chinensis, a specialist on Japanese knotweed Reynoutria japonica. Overall, fungal hyphae and leaf rolls contained significantly higher carbon (C), stable isotopes of C (δ13C), and nitrogen (δ15N) but significantly lower C/N ratios compared to unrolled leaves, whereas insect bodies contained significantly higher N contents but lower C and C/N ratios compared to other types of samples. The MixSIAR model indicated that fungal hyphae contributed a larger proportion (0.626-0.797) to the diet of E. chinensis larvae than leaf materials. The levels of ergosterol, six essential amino acids, seven nonessential amino acids, and three B vitamins tested in fungal hyphae and/or leaf rolls were significantly higher than in unrolled leaves and/or larvae. The P. herquei genome contains the complete set of genes required for the biosynthesis of ergosterol, the essential amino acids valine and threonine, nine nonessential amino acids, and vitamins B2 and B3, whereas some genes associated with five essential and one nonessential amino acid were lost in the P. herquei genome. These suggest that P. herquei is capable of providing the E. chinensis larvae food with ergosterol, amino acids, and B vitamins. P. herquei appears to be able to synthesize or concentrate these nutrients considering that they were specifically concentrated in fungal hyphae. IMPORTANCE: The cultivation of fungi for food has occurred across divergent insect lineages such as social ants, termites, and ambrosia beetles, as well as some seldom-reported solitary insects. Although the fungal cultivars of these insects have been studied for decades, the dietary potential of fungal cultivars for their hosts (especially for those nonsocial insects) is largely unknown. Our research on the mutualistic system Euops chinensis-Penicillium herquei represents an example of the diverse nutritional potentials of the fungal cultivar P. herquei in the diet of the larvae of its solitary host, E. chinensis. These results demonstrate that P. herquei has the potential to synthesize or concentrate ergosterol, amino acids, and B vitamins and benefits the larvae of E. chinensis. Our findings would shed light on poorly understood fungal cultivation mutualisms in nonsocial insects and underscore the nutritional importance of fungal cultivars in fungal cultivation mutualisms.

Physicochemical comparison of chitin characteristics in three major stored-product beetle pests: Implications for biofumigant toxicity.

The present study investigates the chitin properties of stored-product insect pests and their association with the fumigant toxicity of garlic essential oil. Chitin isolates of Callosobruchus maculatus, Sitophilus oryzae, and Tribolium castaneum adults were characterized using FT-IR, XRD, EA, SEM-EDS, and NMR techniques. Fumigant toxicity assay was performed under airtight condition in glass vial. The S. oryzae contains highest chitin content (19 %), followed by T. castaneum (10 %) and C. maculatus (8 %). The degree of crystallinity was lower in C. maculatus (67.13 %) than in S. oryzae (77.05 %) and T. castaneum (76.56 %). Morphologically, C. maculatus chitin displayed a flat lamellar surface with pores, while S. oryzae and T. castaneum exhibited densely arranged microfibrils based surfaces. Fumigant toxicity assays revealed varied susceptibility levels, C. maculatus exhibited higher susceptibility (0.27 µL/L air of LC50) compared to S. oryzae and T. castaneum (14.35 and 3.74 µL/L air of LC50, respectively) to garlic essential oil. The higher chitin content, greater crystallinity, and densely arranged structures in S. oryzae might contribute to its tolerance towards fumigant. Additionally, physico-chemical properties and penetration potentiality of the bioactive constituents might be linked to the toxicity in insects. Understanding these relations can enrich knowledge of chitin's role in fumigant toxicity mechanism.

Exploring the functional profiles of odorant binding proteins crucial for sensing key odorants in the new leaves of coconut palms in Rhynchophorus ferrugineus.

The red palm weevil (RPW), Rhynchophorus ferrugineus (Curculionidae: Coleoptera) is a highly destructive global pest of coconut trees, with a preference for laying its eggs on new leaves. Females can identify where to lay eggs by using their sense of smell to detect specific odorants found in new leaves. In this study, we focused on the two odorants commonly found in new leaves by GC-MS: trans, trans-2,4-nonadienal and trans-2-nonenal. Our behavioral assays demonstrated a significant attraction of females to both of these odorants, with their electrophysiological responses being dose-dependent. Furthermore, we examined the expression patterns induced by these odorants in eleven RferOBP genes. Among them, RferOBP3 and RferOBP1768 exhibited the most significant and simultaneous upregulation. To further understand the role of these two genes, we conducted experiments with females injected with OBP-dsRNA. This resulted in a significant decrease in the expression of RferOBP3 and RferOBP1768, as well as impaired the perception of the two odorants. A fluorescence competitive binding assay also showed that both RferOBPs strongly bound to the odorants. Additionally, sequence analysis revealed that these two RferOBPs belong to the Minus-C family and possess four conserved cysteines. Molecular docking simulations showed strong interactions between these two RferOBPs and the odorant molecules. Overall, our findings highlight the crucial role of RferOBP3 and RferOBP1768 in the olfactory perception of the key odorants in coconut palm new leaves. This knowledge significantly improves our understanding of how RPW females locate sites for oviposition and lays the foundation for future research on the development of environmentally friendly pest attractants.