The genome sequence of a longhorn beetle, Rhagium mordax (Degeer, 1775)
Maxwell V. L. Barclay, Dmitry Telnov, Jay K Goldberg, Bin Zhang
TL;DR
This paper presents the genome sequence of the longhorn beetle Rhagium mordax, including a detailed assembly and gene annotation.
Contribution
The study provides a high-quality genome assembly and gene annotation for Rhagium mordax, a species in the Cerambycidae family.
Findings
The genome assembly is 775.60 megabases long, with 99.53% scaffolded into 10 chromosomal pseudomolecules.
The mitochondrial genome is 16.68 kilobases in length.
Gene annotation identified 11,937 protein-coding genes using Ensembl.
Abstract
We present a genome assembly from an individual female specimen of Rhagium mordax (longhorn beetle; Arthropoda; Insecta; Coleoptera; Cerambycidae). The genome sequence has a total length of 775.60 megabases. Most of the assembly (99.53%) is scaffolded into 10 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 16.68 kilobases in length. Gene annotation of this assembly on Ensembl identified 11,937 protein-coding genes.
Genes, proteins, chemicals, diseases, species, mutations and cell lines named across the full text — each resolved to its canonical identifier and authoritative record.
Click any figure to enlarge with its caption.
Figure 1
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Figure 5| Project information | |||
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| PRJEB63422 | ||
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| SAMEA111458823 | ||
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| Specimen information | |||
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| icRhaMord1 | SAMEA111458864 | Whole organism |
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| icRhaMord1 | SAMEA111458823 | Whole organism |
| Sequencing information | |||
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| ERR11606307 | 6.27e+08 | 94.64 |
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| ERR11593795 | 2.31e+06 | 21.8 |
| Genome assembly | ||
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| Assembly name | icRhaMord1.1 | |
| Assembly accession | GCA_963680705.1 | |
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| Span (Mb) | 775.60 | |
| Number of contigs | 515 | |
| Number of scaffolds | 68 | |
| Longest scaffold (Mb) | 134.33 | |
| Assembly metrics
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| Contig N50 length (Mb) | 2.9 |
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| Scaffold N50 length (Mb) | 80.1 |
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| Consensus quality (QV) | Primary: 61.2; alternate: 60.6; combined 60.9 |
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| Primary: 86.98%; alternate: 72.20%; combined:
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| BUSCO v5.4.3 lineage:
| C:99.0%[S:97.8%,D:1.2%],
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| Percentage of assembly mapped to chromosomes | 99.53% |
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| Sex chromosomes | Not identified |
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| Organelles | Mitochondrial genome: 16.68 kb |
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| Genome annotation of assembly GCA_963680705.1 at Ensembl | ||
| Number of protein-coding genes | 11,937 | |
| Number of non-coding genes | 1,505 | |
| Number of gene transcripts | 19,101 | |
| INSDC
| Name | Length
| GC% |
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| 1 | 134.33 | 32.5 | |
| 2 | 110.84 | 32.5 | |
| 3 | 94.63 | 32.5 | |
| 4 | 80.11 | 32.5 | |
| 5 | 76.39 | 32.5 | |
| 6 | 76.33 | 33.0 | |
| 7 | 60.86 | 33.0 | |
| 8 | 54.87 | 34.0 | |
| 9 | 47.15 | 34.5 | |
| 10 | 35.99 | 34.0 | |
| MT | 0.02 | 20.5 |
| Software
| Version | Source |
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| BEDTools | 2.30.0 |
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| BLAST | 2.14.0 |
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| BlobToolKit | 4.3.7 |
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| BUSCO | 5.4.3 and 5.5.0 |
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| bwa-mem2 | 2.2.1 |
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| Cooler | 0.8.11 |
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| DIAMOND | 2.1.8 |
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| fasta_
| 0.2.4 |
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| FastK | 427104ea91c78c3b8b8b49f1a7d6bbeaa869ba1c |
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| Gfastats | 1.3.6 |
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| GoaT CLI | 0.2.5 |
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| Hifiasm | 0.19.8-r587 |
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| HiGlass | 44086069ee7d4d3f6f3f0012569789ec138f42b84a
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| Merqury.FK | d00d98157618f4e8d1a9190026b19b471055b22e |
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| MitoHiFi | 3 |
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| MultiQC | 1.14, 1.17, and 1.18 |
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| NCBI
| 15.12.0 |
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| Nextflow | 23.04.0-5857 |
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| PretextView | 0.2.5 |
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| purge_
| 1.2.5 |
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| samtools | 1.16.1, 1.17, and 1.18 |
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| sanger-tol/
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| Seqtk | 1.3 |
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| Singularity | 3.9.0 |
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| TreeVal | 1.0.0 |
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| YaHS | 1.2a.2 |
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- —Wellcome Trust
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Taxonomy
TopicsForest Insect Ecology and Management · Forest Ecology and Biodiversity Studies · Coleoptera Taxonomy and Distribution
Species taxonomy
Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Protostomia; Ecdysozoa; Panarthropoda; Arthropoda; Mandibulata; Pancrustacea; Hexapoda; Insecta; Dicondylia; Pterygota; Neoptera; Endopterygota; Coleoptera; Polyphaga; Cucujiformia; Chrysomeloidea; Cerambycidae; Lepturinae; Rhagiini; Rhagium; Rhagium mordax (Degeer, 1775) (NCBI:txid295679)
Background
The Cerambycidae Latreille, 1802, commonly known as longhorned beetles, longicorns, capricorns, round-headed borers, timber beetles, or sawyer beetles, comprises one of the largest families of the order Coleoptera with at least 35,000 species worldwide ( Rossa & Goczał, 2021). Moreover, cerambycids represents one of most diverse, ecologically, and economically important group of beetles. The family is of cosmopolitan distribution, most speciose in the tropical regions. Longhorn beetle larvae are usually saproxylic, xylophagous or herbicolous, feed on decaying or live plant tissues and provide essential ecosystem service by rapid recycling dead wood into humus. Adults are phytophagous, feeding on green or withered plants, many species are anthophilous or forage for fermented tree sap. The English names of the family refer to the elongate antennae of these beetles or to their connection with wood. Nine extant subfamilies are recognized within Cerambycidae ( Bouchard et al., 2011). In the British fauna, there are 68 established species of longhorn beetles known presently ( Duff, 2018). The genus Rhagium Fabricius, 1775 is represented in the Palaearctic Region by three subgenera and 26 species and subspecies ( Danilevsky, 2020). Seven species and subspecies of Rhagium occur in Europe ( Danilevsky, 2020), three of which are present in the British fauna ( Duff, 2020).
Rhagium mordax (DeGeer, 1775) is placed in the subfamily Lepturinae tribe Rhagiini ( Danilevsky, 2020). The species is assigned to the subgenus Megarhagium Reitter, 1913. Adults of the species are well-defined morphologically and resemble only those of R. (M.) sycophanta (Schrank, 1781), a slightly larger congener which does not occur in Britain (although erroneously listed as British in Danilevsky, 2020). Paulus (1969) provided a key to the Central European Rhagium larvae. It is hoped that genomic data may help clarify the identification of immature Rhagium.
Rhagium mordax is a trans-Palaearctic species widely distributed from northern and eastern Spain and the British Isles towards the eastern Siberia and Kazakhstan ( Danilevsky, 2020 and references therein; GBIF Secretariat, 2024). Danilevsky (2020) list the species from 37 European countries, including the UK. Generally, it is much less abundant in southern and southeastern Europe than in the zone of temperate and boreal forests. In Asia, the species is distributed in western and eastern Siberia, northern Kazakhstan, with an isolated subpopulation in Turkmenistan ( Danilevsky, 2020 and references therein). The European extent of occurrence and area of occupancy of this species are both strongly above the thresholds for a threatened species ( Dodelin et al., 2017).
Rhagium mordax is a forest species. Its larvae are saproxylic and polyphagous, developing for 2–3 years between bark and hardwood of decaying Alnus spp., Betula spp., Fagus sylvatica, Populus tremula, Quercus spp., occasionally also coniferous trees ( Duff, 2016; Koch, 1992; Nikitsky et al., 1996; Palm, 1959). Larval development takes two years in the northern and eastern parts of the range, pupae are found throughout the year ( Nikitsky et al., 1996; Palm, 1959). This species uses wood in a broad range of conditions of for larval development, including snags and fallen trunks in shady or sun exposed, wet or moderately dry conditions. Although generally associated with deciduous trees, live larvae have been collected under bark of pine Pinus sp. washed up on a beach in North Devon (M.V.L. Barclay personal observation). The species is described as eurytopic, silvicol, corticol, lignicol, xylophagous and anthophilous ( Koch, 1992). Adult beetles occur in forests, on forest edges, hedgerows and clearing. They feed on pollen of various blossoming plants, in Britain most often the blossom of hawthorn Crataegus (Rosaceae); they have also been recorded feeding on fermented tree sap (D. Telnov, personal observations in Latvia).
The sequenced larva was sampled in April from under bark of a dead and decaying oak trunk Quercus robur. In Britain, adults have been found all year ( Duff, 2016), but mostly inside the larval substrate, becoming active outside the wood in Spring and early Summer. In the northern and eastern parts of the distribution area, the adults appear in July to August.
Rhagium mordax is a common and widespread species in United Kingdom and recorded in England, Wales and Scotland northwards to West Sutherland as well as in Ireland ( Duff, 2016). The species was not listed in the national Red Data Book ( Shirt, 1987), the present status of the species in the United Kingdom is Least Concern (LC) ( Alexander, 2014).
The specimen used for sequencing, a mature larva under oak bark at Bookham Common, Surrey, southern England, was collected and identified by M.V.L. Barclay.
Genome sequence report
Sequencing data
The genome of Rhagium mordax ( Figure 1) was sequenced using Pacific Biosciences single-molecule HiFi long reads, generating a total of 21.80 Gb (gigabases) from 2.31 million reads, providing an estimated 26-fold coverage. Hi-C sequencing produced 94.64 Gb from 626.77 million reads. Specimen and sequencing details are summarised in Table 1.
Photographs of the Rhagium mordax (icRhaMord1) specimen used for genome sequencing.
Table 1.: Specimen and sequencing data for Rhagium mordax.
Assembly statistics
The primary haplotype was assembled, and contigs corresponding to an alternate haplotype were also deposited in INSDC databases. The assembly was improved by manual curation, which corrected 197 misjoins or missing joins and removed 47 haplotypic duplications. These interventions reduced the total assembly length by 1.82%, decreased the scaffold count by 47.73%, and increased the scaffold N50 by 0.56%. The final assembly has a total length of 775.64 Mb in 68 scaffolds, with 446 gaps, and a scaffold N50 of 80.11 Mb ( Table 2).
Table 2.: Genome assembly data for Rhagium mordax, icRhaMord1.1.
The snail plot in Figure 2 provides a summary of the assembly statistics, indicating the distribution of scaffold lengths and other assembly metrics. Figure 3 shows the distribution of scaffolds by GC proportion and coverage. Figure 4 presents a cumulative assembly plot, with separate curves representing different scaffold subsets assigned to various phyla, illustrating the completeness of the assembly.
Genome assembly of Rhagium mordax, icRhaMord1.1: metrics.The BlobToolKit snail plot provides an overview of assembly metrics and BUSCO gene completeness. The circumference represents the length of the whole genome sequence, and the main plot is divided into 1,000 bins around the circumference. The outermost blue tracks display the distribution of GC, AT, and N percentages across the bins. Scaffolds are arranged clockwise from longest to shortest and are depicted in dark grey. The longest scaffold is indicated by the red arc, and the deeper orange and pale orange arcs represent the N50 and N90 lengths. A light grey spiral at the centre shows the cumulative scaffold count on a logarithmic scale. A summary of complete, fragmented, duplicated, and missing BUSCO genes in the endopterygota_odb10 set is presented at the top right. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/GCA_963680705.1/dataset/GCA_963680705.1/snail.
Genome assembly of Rhagium mordax, icRhaMord1.1: BlobToolKit GC-coverage plot showing sequence coverage (vertical axis) and GC content (horizontal axis).The circles represent scaffolds, with the size proportional to scaffold length and the colour representing phylum membership. The histograms along the axes display the total length of sequences distributed across different levels of coverage and GC content. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/GCA_963680705.1/dataset/GCA_963680705.1/blob.
Genome assembly of Rhagium mordax icRhaMord1.1: BlobToolKit cumulative sequence plot.The grey line shows cumulative length for all scaffolds. Coloured lines show cumulative lengths of scaffolds assigned to each phylum using the buscogenes taxrule. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/GCA_963680705.1/dataset/GCA_963680705.1/cumulative.
Most of the assembly sequence (99.47%) was assigned to 10 chromosomal-level scaffolds. These chromosome-level scaffolds, confirmed by Hi-C data, are named according to size ( Figure 5; Table 3). During curation, it was noted that the sample is homogametic sex (female) but X chromosome could not be identified.
Genome assembly of Rhagium mordax icRhaMord1.1: Hi-C contact map of the icRhaMord1.1 assembly, visualised using HiGlass.Chromosomes are shown in order of size from left to right and top to bottom. An interactive version of this figure may be viewed at https://genome-note-higlass.tol.sanger.ac.uk/l/?d=YRLlDEwzQjiwvrfgTh3rVA.
Table 3.: Chromosomal pseudomolecules in the genome assembly of Rhagium mordax, icRhaMord1.
The mitochondrial genome was also assembled. This sequence is included as a contig in the multifasta file of the genome submission, and as a separate fasta file with accession OY796666.1.
Assembly quality metrics
The estimated Quality Value (QV) and k-mer completeness metrics, along with BUSCO completeness scores, were calculated for each haplotype and the combined assembly. The QV reflects the base-level accuracy of the assembly, while k-mer completeness indicates the proportion of expected k-mers identified in the assembly. BUSCO scores provide a measure of completeness based on benchmarking universal single-copy orthologues.
The primary haplotype has a QV of 61.2, and the combined primary and alternate assemblies achieve an estimated QV of 60.9. The k-mer completeness for the primary haplotype is 86.98%, and for the alternate haplotype it is 72.20%. The combined primary and alternate assemblies achieve a k-mer completeness of 97.79%. BUSCO analysis using the endopterygota_odb10 reference set ( n = 2,124) indicated a completeness score of 99.0% (single = 97.8%, duplicated = 1.2%).
Table 2 provides assembly metric benchmarks adapted from Rhie et al. (2021) and the Earth BioGenome Project Report on Assembly Standards September 2024. The assembly achieves the EBP reference standard of 6.C.61.
Genome annotation report
The Rhagium mordax genome assembly (GCA_963680705.1) was annotated at the European Bioinformatics Institute (EBI) on Ensembl Rapid Release. The resulting annotation includes 19,101 transcribed mRNAs from 11,937 protein-coding and 1,505 non-coding genes ( Table 2; https://rapid.ensembl.org/Rhagium_mordax_GCA_963680705.1/Info/Index). The average transcript length is 12,324.32. There are 1.42 coding transcripts per gene and 5.16 exons per transcript.
Methods
Sample acquisition and DNA barcoding
A female larval specimen of Rhagium mordax (specimen ID NHMUK014433257, ToLID icRhaMord1) was collected from Bookham Common, England, United Kingdom (latitude 51.29, longitude –0.39) on 2021-04-18. The specimen was collected and identified by Maxwell Barclay (Natural History Museum) and preserved by dry freezing at –80 °C.
The initial identification was verified by an additional DNA barcoding process according to the framework developed by Twyford et al. (2024). A small sample was dissected from the specimens and stored in ethanol, while the remaining parts were shipped on dry ice to the Wellcome Sanger Institute (WSI). The tissue was lysed, the COI marker region was amplified by PCR, and amplicons were sequenced and compared to the BOLD database, confirming the species identification ( Crowley et al., 2023). Following whole genome sequence generation, the relevant DNA barcode region was also used alongside the initial barcoding data for sample tracking at the WSI ( Twyford et al., 2024). The standard operating procedures for Darwin Tree of Life barcoding have been deposited on protocols.io ( Beasley et al., 2023).
Nucleic acid extraction
The workflow for high molecular weight (HMW) DNA extraction at the Wellcome Sanger Institute (WSI) Tree of Life Core Laboratory includes a sequence of procedures: sample preparation and homogenisation, DNA extraction, fragmentation and purification. Detailed protocols are available on protocols.io ( Denton et al., 2023b). The icRhaMord1 sample was prepared for DNA extraction by weighing and dissecting it on dry ice ( Jay et al., 2023). Tissue was homogenised using a PowerMasher II tissue disruptor ( Denton et al., 2023a).
HMW DNA was extracted in the WSI Scientific Operations core using the Automated MagAttract v2 protocol ( Oatley et al., 2023). The DNA was sheared into an average fragment size of 12–20 kb in a Megaruptor 3 system ( Bates et al., 2023). Sheared DNA was purified by solid-phase reversible immobilisation, using AMPure PB beads to eliminate shorter fragments and concentrate the DNA ( Strickland et al., 2023). The concentration of the sheared and purified DNA was assessed using a Nanodrop spectrophotometer and Qubit Fluorometer using the Qubit dsDNA High Sensitivity Assay kit. Fragment size distribution was evaluated by running the sample on the FemtoPulse system.
Hi-C preparation
Tissue from the icRhaMord1 sample was processed at the WSI Scientific Operations core, using the Arima-HiC v2 kit. Tissue (stored at –80 °C) was fixed, and the DNA crosslinked using a TC buffer with 22% formaldehyde. After crosslinking, the tissue was homogenised using the Diagnocine Power Masher-II and BioMasher-II tubes and pestles. Following the kit manufacturer's instructions, crosslinked DNA was digested using a restriction enzyme master mix. The 5’-overhangs were then filled in and labelled with biotinylated nucleotides and proximally ligated. An overnight incubation was carried out for enzymes to digest remaining proteins and for crosslinks to reverse. A clean up was performed with SPRIselect beads prior to library preparation.
Library preparation and sequencing
Library preparation and sequencing were performed at the WSI Scientific Operations core. Pacific Biosciences HiFi circular consensus DNA sequencing libraries were prepared using the PacBio Express Template Preparation Kit v2.0 (Pacific Biosciences, California, USA) as per the manufacturer's instructions. The kit includes the reagents required for removal of single-strand overhangs, DNA damage repair, end repair/A-tailing, adapter ligation, and nuclease treatment. Library preparation also included a library purification step using AMPure PB beads (Pacific Biosciences, California, USA) and size selection step to remove templates shorter than 3 kb using AMPure PB modified SPRI. DNA concentration was quantified using the Qubit Fluorometer v2.0 and Qubit HS Assay Kit and the final library fragment size analysis was carried out using the Agilent Femto Pulse Automated Pulsed Field CE Instrument and gDNA 165kb gDNA and 55kb BAC analysis kit. Samples were sequenced using the Sequel IIe system (Pacific Biosciences, California, USA). The concentration of the library loaded onto the Sequel IIe was in the range 40–135 pM. The SMRT link software, a PacBio web-based end-to-end workflow manager, was used to set-up and monitor the run, as well as perform primary and secondary analysis of the data upon completion.
For Hi-C library preparation, DNA was fragmented to a size of 400 to 600 bp using a Covaris E220 sonicator. The DNA was then enriched, barcoded, and amplified using the NEBNext Ultra II DNA Library Prep Kit following manufacturers’ instructions. The Hi-C sequencing was performed using paired-end sequencing with a read length of 150 bp on an Illumina NovaSeq 6000 instrument.
Genome assembly, curation and evaluation
** Assembly **
The HiFi reads were first assembled using Hifiasm ( Cheng et al., 2021) with the --primary option. Haplotypic duplications were identified and removed using purge_dups ( Guan et al., 2020). The Hi-C reads were mapped to the primary contigs using bwa-mem2 ( Vasimuddin et al., 2019). The contigs were further scaffolded using the provided Hi-C data ( Rao et al., 2014) in YaHS ( Zhou et al., 2023) using the --break option for handling potential misassemblies. The scaffolded assemblies were evaluated using Gfastats ( Formenti et al., 2022), BUSCO ( Manni et al., 2021) and MERQURY.FK ( Rhie et al., 2020).
The mitochondrial genome was assembled using MitoHiFi ( Uliano-Silva et al., 2023), which runs MitoFinder ( Allio et al., 2020) and uses these annotations to select the final mitochondrial contig and to ensure the general quality of the sequence.
** Assembly curation **
The assembly was decontaminated using the Assembly Screen for Cobionts and Contaminants (ASCC) pipeline (article in preparation). Flat files and maps used in curation were generated in TreeVal ( Pointon et al., 2023). Manual curation was primarily conducted using PretextView ( Harry, 2022), with additional insights provided by JBrowse2 ( Diesh et al., 2023) and HiGlass ( Kerpedjiev et al., 2018). Scaffolds were visually inspected and corrected as described by Howe et al. (2021). Any identified contamination, missed joins, and mis-joins were corrected, and duplicate sequences were tagged and removed. The curation process is documented at https://gitlab.com/wtsi-grit/rapid-curation (article in preparation).
** Assembly quality assessment **
The Merqury.FK tool ( Rhie et al., 2020), run in a Singularity container ( Kurtzer et al., 2017), was used to evaluate k-mer completeness and assembly quality for the primary and alternate haplotypes using the k-mer databases ( k = 31) that were computed prior to genome assembly. The analysis outputs included assembly QV scores and completeness statistics.
A Hi-C contact map was produced for the final version of the assembly. The Hi-C reads were aligned using bwa-mem2 ( Vasimuddin et al., 2019) and the alignment files were combined using SAMtools ( Danecek et al., 2021). The Hi-C alignments were converted into a contact map using BEDTools ( Quinlan & Hall, 2010) and the Cooler tool suite ( Abdennur & Mirny, 2020). The contact map is visualised in HiGlass ( Kerpedjiev et al., 2018).
The blobtoolkit pipeline is a Nextflow port of the previous Snakemake Blobtoolkit pipeline ( Challis et al., 2020). It aligns the PacBio reads in SAMtools and minimap2 ( Li, 2018) and generates coverage tracks for regions of fixed size. In parallel, it queries the GoaT database ( Challis et al., 2023) to identify all matching BUSCO lineages to run BUSCO ( Manni et al., 2021). For the three domain-level BUSCO lineages, the pipeline aligns the BUSCO genes to the UniProt Reference Proteomes database ( Bateman et al., 2023) with DIAMOND blastp ( Buchfink et al., 2021). The genome is also divided into chunks according to the density of the BUSCO genes from the closest taxonomic lineage, and each chunk is aligned to the UniProt Reference Proteomes database using DIAMOND blastx. Genome sequences without a hit are chunked using seqtk and aligned to the NT database with blastn ( Altschul et al., 1990). The blobtools suite combines all these outputs into a blobdir for visualisation.
The blobtoolkit pipeline was developed using nf-core tooling ( Ewels et al., 2020) and MultiQC ( Ewels et al., 2016), relying on the Conda package manager, the Bioconda initiative ( Grüning et al., 2018), the Biocontainers infrastructure ( da Veiga Leprevost et al., 2017), as well as the Docker ( Merkel, 2014) and Singularity ( Kurtzer et al., 2017) containerisation solutions.
Table 4 contains a list of relevant software tool versions and sources.
Genome annotation
The Ensembl Genebuild annotation system ( Aken et al., 2016) was used to generate annotation for the Rhagium mordax assembly (GCA_963680705.1) in Ensembl Rapid Release at the EBI. Annotation was created primarily through alignment of transcriptomic data to the genome, with gap filling via protein-to-genome alignments of a select set of proteins from UniProt ( UniProt Consortium, 2019).
Wellcome Sanger Institute – Legal and Governance
The materials that have contributed to this genome note have been supplied by a Darwin Tree of Life Partner. The submission of materials by a Darwin Tree of Life Partner is subject to the ‘Darwin Tree of Life Project Sampling Code of Practice’, which can be found in full on the Darwin Tree of Life website here. By agreeing with and signing up to the Sampling Code of Practice, the Darwin Tree of Life Partner agrees they will meet the legal and ethical requirements and standards set out within this document in respect of all samples acquired for, and supplied to, the Darwin Tree of Life Project.
Further, the Wellcome Sanger Institute employs a process whereby due diligence is carried out proportionate to the nature of the materials themselves, and the circumstances under which they have been/are to be collected and provided for use. The purpose of this is to address and mitigate any potential legal and/or ethical implications of receipt and use of the materials as part of the research project, and to ensure that in doing so we align with best practice wherever possible. The overarching areas of consideration are:
• Ethical review of provenance and sourcing of the material
• Legality of collection, transfer and use (national and international)
Each transfer of samples is further undertaken according to a Research Collaboration Agreement or Material Transfer Agreement entered into by the Darwin Tree of Life Partner, Genome Research Limited (operating as the Wellcome Sanger Institute), and in some circumstances other Darwin Tree of Life collaborators.
The reference list from the paper itself. Each links out to its DOI / PubMed record.
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- 3Alexander KNA : A review of the status of the beetles of Great Britain. Longhorn Beetles (Cerambycidae). Species Status No. 39. Natural England Commissioned Report NECR 148,2014. Reference Source
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- 5Altschul SF Gish W Miller W : Basic local alignment search tool. J Mol Biol. 1990;215(3):403–410. 10.1016/S 0022-2836(05)80360-2 2231712 · doi ↗ · pubmed ↗
- 6Bateman A Martin MJ Orchard S : Uni Prot: the Universal Protein Knowledgebase in 2023. Nucleic Acids Res. 2023;51(D 1):D 523–D 531. 10.1093/nar/gkac 1052 36408920 PMC 9825514 · doi ↗ · pubmed ↗
- 7Bates A Clayton-Lucey I Howard C : Sanger Tree of Life HMW DNA fragmentation: diagenode Megaruptor ®3 for LI Pac Bio. protocols.io. 2023. 10.17504/protocols.io.81wgbxzq 3lpk/v 1 · doi ↗
- 8Beasley J Uhl R Forrest LL : DNA barcoding SO Ps for the Darwin Tree of Life project. protocols.io. 2023; [Accessed 25 June 2024]. 10.17504/protocols.io.261ged 91jv 47/v 1 · doi ↗