Abstract
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.
Highlights
The large pine weevil, Hylobius abietis, (Linnaeus Coleoptera Curculionidae), is a notorious pest of coniferous trees of genus Pinus
The neuropeptide profile of H. abietis from a combined approach of transcriptomics and peptidomics features the most complete repertoire of insect neuropeptides identified in a beetle to date and without the need for sequencing the whole genome. 41 different insect neuropeptide families were identified from 52 neuropeptide precursor sequences
Allatostatin A, Allatostatin CCC, Corazonin, Kinin, Sex Peptide and Pigment Dispersing Factor mature peptides were not identified in H. abietis, consistent with T. castaneum
Summary
The large pine weevil, Hylobius abietis, (Linnaeus Coleoptera Curculionidae), is a notorious pest of coniferous trees of genus Pinus. Coleoptera is the most-species rich Order of Class Insecta and is a very diverse group consisting of many harmful pests as well as a small percentage of beneficial ones (Riley and Goyer, 1986; Teshler et al, 2004; Nash et al, 2008). Due to their agricultural and economical importance, there has been a progressive increase in the study of beetles at the gene level, as evidenced by the genome sequencing of several
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