Coordinated RNA-Seq and peptidomics identify neuropeptides and G-protein coupled receptors (GPCRs) in the large pine weevil Hylobius abietis, a major forestry pest

Hylobius abietis (Linnaeus), or large pine weevil (Coleoptera, Curculionidae), is a pest of European coniferous forests. In order to gain understanding of the functional physiology of this species, we have assembled a de novo transcriptome of H. abietis, from sequence data obtained by Next Generation Sequencing. In particular, we have identified genes encoding neuropeptides, peptide hormones and their putative G-protein coupled receptors (GPCRs) to gain insights into neuropeptide-modulated processes. The transcriptome was assembled de novo from pooled paired-end, sequence reads obtained from RNA from whole adults, gut and central nervous system tissue samples. Data analysis was performed on the transcripts obtained from the assembly including, annotation, gene ontology and functional assignment as well as transcriptome completeness assessment and KEGG pathway analysis. Pipelines were created using Bioinformatics tools and techniques for prediction and identification of neuropeptides and neuropeptide receptors. Peptidomic analysis was also carried out using a combination of MALDI-TOF as well as Q-Exactive Orbitrap mass spectrometry to confirm the identified neuropeptide. 41 putative neuropeptide families were identified in H. abietis, including Adipokinetic hormone (AKH), CAPA and DH31. Neuropeptide F, which has not been yet identified in the model beetle T. castaneum, was identified. Additionally, 24 putative neuropeptide and 9 leucine-rich repeat containing G protein coupled receptor-encoding transcripts were determined using both alignment as well as non-alignment methods. This information, submitted to the NCBI sequence read archive repository (SRA accession: SRP133355), can now be used to inform understanding of neuropeptide-modulated physiology and behaviour in H. abietis; and to develop specific neuropeptide-based tools for H. abietis control

Predicting the unpredictable: How host specific is the mycobiota of bark and ambrosia beetles?

Bark and ambrosia beetles (Curculionidae: Scolytinae) are known for their symbioses with fungi and play a key role in the dispersal of phytopathogens. The scolytine community of eight pine stands along a latitudinal gradient in the UK was surveyed and beetle-associated fungal communities (mycobiota) were assessed using ITS2 metabarcoding (304 specimens, 12 species). Distribution patterns among 2,257 detected fungal Operational Taxonomic Units (OTUs) revealed that beetle species identity was an important predictor of mycobiotic richness and composition, while the effects of environmental and spatial variables were negligible. Network-based specificity analysis suggested that a relatively small subset of OTUs (75 in total) exhibit an affinity for a subset of beetle species and that these include many Microascales and Saccharomycetes. Notably though, of the OTUs belonging to the family Ophiostoma-taceae, relatively few display host specificity. Our results add to the complex picture of host-associated fungal communities and suggest that host range limits are unlikely to restrict the spread of economically important phytopathogens. Crown Copyright (C) 2019 Published by Elsevier Ltd. All rights reserved

Evidence of cross-channel dispersal into England of the forest pest Ips typographus

Abstract A breeding population of the tree-killing European spruce bark beetle Ips typographus was detected in England for the first time in 2018 and was initially assumed to have arrived with infested timber. To test the hypothesis that the beetles are dispersing naturally across the English channel, extensive trap networks were deployed in 2021 and 2022 to track the flight activity of the beetles from an outbreak hotspot in France and Belgium to southern England, including parallel ‘coastal’ traps on either side of the channel. Beetles were caught all along the transect, decreasing in abundance with distance from the outbreak area. Linear modelling indicates that beetles dispersed into England during 2021 and 2022, and that during a large-scale dispersal event in June 2021, beetles could have penetrated more than 160 km inland. The 2021 dispersal event initiated new incursions of the beetle in southeast England and demonstrates the extraordinary distance I. typographus may move under outbreak conditions. Our findings support the hypothesis of a damaging forest pest aerially dispersing across the barrier of the English channel and suggest that future incursions of this and other plant-associated pests may move via the same pathway.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Cosmopolitan Scolytinae: strong common drivers, but too many singularities for accurate prediction

Many scolytine beetle species have been expanding in new territories, travelling with wood and plants for planting, sometimes with a high impact on plant health. Here, we attempt to quantify the mobility of these species and to identify the biological drivers of mobility and impact. Mobility was estimated by counting the numbers of landmasses (contiguous pieces of land, surrounded by ocean or sea) colonised by each species. A series of potential drivers (taxonomic tribes; feeding regimes; polyphagy; reproductive strategy; host taxa; aggregation pheromones and long-range primary attractants), as well as impact on host health were recorded. A total of 163 species were identified, out of 5546 counted in the whole subfamily. The cosmopolitan taxa amongst the subfamily showed significant disharmony with regards to invasion frequency. Four tribes (Xyleborini; Ipini; Crypturgini; Hylastini) were significantly over-represented and two others (Corthylini; Hexacolini) were under-represented. Some 53% of the 163 species are inbreeding, a very significant excess as compared to the whole subfamily (29%). The inbreeders colonised more landmasses than the outbreeders. There is a significant relationship between the number of host families attacked by a species and the number of colonised landmasses. Most of the invasive species are recorded to respond to long-range host primary attractants, only one quarter respond to pheromones. All very mobile species respond to long-range primary attractants and none is known to respond to pheromones. Very mobile species are all associated with a substantial or moderate impact. The most mobile species belong to a limited number of subtribes. They are often inbreeding, polyphagous and respond to long-range primary attractants, but do not produce pheromones. However, there are many counter-examples. The outbreeding Scolytus multistriatus attacks only three host families, producing aggregation pheromones and has established in thirteen landmasses, with a high impact. Due to these many exceptions, species-based risk prediction relying on the few traits routinely analysed in literature suffers from important uncertainties.info:eu-repo/semantics/publishe

Death of an order: a comprehensive molecular phylogenetic study confirms that termites are eusocial cockroaches

Termites are instantly recognizable mound-builders and house-eaters: their complex social lifestyles have made them incredibly successful throughout the tropics. Although known as ‘white ants’, they are not ants and their relationships with other insects remain unclear. Our molecular phylogenetic analyses, the most comprehensive yet attempted, show that termites are social cockroaches, no longer meriting being classified as a separate order (Isoptera) from the cockroaches (Blattodea). Instead, we propose that they should be treated as a family (Termitidae) of cockroaches. It is surprising to find that a group of wood-feeding cockroaches has evolved full sociality, as other ecologically dominant fully social insects (e.g. ants, social bees and social wasps) have evolved from solitary predatory wasps

Metabarcoding of insect-associated fungal communities: a comparison of internal transcribed spacer (ITS) and large-subunit (LSU) rRNA markers

Full taxonomic characterisation of fungal communities is necessary for establishing ecological associations and early detection of pathogens and invasive species. Complex communities of fungi are regularly characterised by metabarcoding using the Internal Transcribed Spacer (ITS) and the Large-Subunit (LSU) gene of the rRNA locus, but reliance on a single short sequence fragment limits the confidence of identification. Here we link metabarcoding from the ITS2 and LSU D1-D2 regions to characterise fungal communities associated with bark beetles (Scolytinae), the likely vectors of several tree pathogens. Both markers revealed similar patterns of overall species richness and response to key variables (beetle species, forest type), but identification against the respective reference databases using various taxonomic classifiers revealed poor resolution towards lower taxonomic levels, especially the species level. Thus, Operational Taxonomic Units (OTUs) could not be linked via taxonomic classifiers across ITS and LSU fragments. However, using phylogenetic trees (focused on the epidemiologically important Sordariomycetes) we placed OTUs obtained with either marker relative to reference sequences of the entire rRNA cistron that includes both loci and demonstrated the largely similar phylogenetic distribution of ITS and LSU-derived OTUs. Sensitivity analysis of congruence in both markers suggested the biologically most defensible threshold values for OTU delimitation in Sordariomycetes to be 98% for ITS2 and 99% for LSU D1-D2. Studies of fungal communities using the canonical ITS barcode require corroboration across additional loci. Phylogenetic analysis of OTU sequences aligned to the full rRNA cistron shows higher success rate and greater accuracy of species identification compared to probabilistic taxonomic classifiers

The risk of bark and ambrosia beetles associated with imported non-coniferous wood and potential horizontal phytosanitary measures

Many bark and ambrosia beetle species (Coleoptera: Scolytinae and Platypodinae) are known to have spread worldwide in relation to international trade. Concerns have been expressed within the European and Mediterranean Plant Protection Organization (EPPO) about recent introductions of non-indigenous species of these groups. Regulation of the non-coniferous wood trade into many EPPO member countries is currently not sufficient to cover such risks. In 2018-2019, an EPPO study on the risk of bark and ambrosia beetles associated with imported non-coniferous wood was carried out, and the key characteristics contributing to the pest risk from introduced species were determined using expert consensus. This paper summarizes the key findings of the study, which are available in full detail on the EPPO website. The study identified biological and other risk factors and illustrated them with examples from 26 beetle species or groups of species known to be invasive or posing a threat to plant health. These representative species were classified into three categories based on known damage and level of uncertainty. In the present article, factorial discriminant analyses were used to identify features of bark and ambrosia beetle biology associated with damage caused and invasiveness. Based on the information assembled and consideration of the risk factors, it was recommended that in order to prevent the introduction of new bark and ambrosia beetles via non-coniferous wood commodities, horizontal phytosanitary measures should be adopted, irrespective of the host plant species and the origin (i.e., for all genera of non-coniferous woody plants and from all origins). Phytosanitary measures are presented here for various wood commodities