The genome sequence of the Figure of Eighty moth Tethea ocularis Linnaeus, 1767

We present a genome assembly from an individual male Tethea ocularis (the Figure of Eighty; Arthropoda; Insecta; Lepidoptera; Drepanidae). The genome sequence is 339.1 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.28 kilobases in length


Background
Tethea ocularis the Figure of Eighty is a moth in the family Drepanidae, in a group known as lutestrings that was formerly the separate family Thyatiridae.The Drepanidae look similar to the noctuid moths, although they are not as thick-bodied.Tethea ocularis has a brown forewing, varying in shade from light to dark brown, fine crosslines and a distinctive white "80" in the centre of the forewing: it is superficially similar to the Poplar Lutestring Tethea or and the noctuid We present a chromosomally complete genome sequence for an adult male Tethea ocularis, based on one specimen collected in Wytham Woods, Oxfordshire, as part of the Darwin Tree of Life Project.

Genome sequence report
The genome was sequenced from a male Tethea ocularis (Figure 1) collected from Wytham Woods, Oxfordshire, UK (51.77, -1.34).A total of 223-fold coverage in Pacific Biosciences single-molecule HiFi long reads was generated.Primary assembly contigs were scaffolded with chromosome conformation Hi-C data.Manual assembly curation corrected 10 missing joins or mis-joins, increasing the scaffold number by 1.69%.
The final assembly has a total length of 339.1 Mb in 59 sequence scaffolds with a scaffold N50 of 12.1 Mb (Table 1).The snail plot in Figure 2 provides a summary of the assembly statistics, while the distribution of assembly scaffolds on GC proportion and coverage is shown in Figure 3.The cumulative assembly plot in Figure 4 shows curves for subsets of scaffolds assigned to different phyla.Most (99.6%) of the assembly sequence was assigned to 31 chromosomal-level scaffolds, representing 30 autosomes and the Z sex chromosome.Chromosome-scale scaffolds confirmed by the Hi-C data are named in order of size (Figure 5; Table 2).The Z sex chromosome was assigned by synteny to Tetheella fluctuosa (GCA_951216915.1).While not fully phased, the assembly deposited is of one haplotype.Contigs corresponding to the second haplotype have also been deposited.The mitochondrial genome was also assembled and can be found as a contig within the multifasta file of the genome submission.

Sample acquisition and nucleic acid extraction
A male Tethea ocularis (specimen ID Ox001919, ToLID ilTetOcul2) was collected from Wytham Woods, Oxfordshire (biological vice-county Berkshire), UK (latitude 51.77, longitude -1.34) on 2021-06-16.The specimen used for Hi-C and RNA sequencing (specimen ID Ox000465, ToLID ilTetOcul1) was collected from the same location on 2020-06-13.Both specimens were collected and identified by Douglas Boyes (University of Oxford) and preserved on dry ice.
The workflow for high molecular weight (HMW) DNA extraction at the Wellcome Sanger Institute (WSI) includes a sequence of HMW DNA was extracted in the WSI Scientific Operations core using the Automated MagAttract v2 protocol   Table 3 contains a list of relevant software tool versions and sources.

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   Table 3. Software tools: versions and sources.

Software tool Version
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

Panagiotis Ioannidis
Foundation for Research & Technology -Hellas, Heraklion, Crete, GR 70013, Greece The herein reported genome sequence of the Tethea ocularis moth is a high-quality assembly which is based on deep, PacBio sequencing (223-fold coverage).Additionally, BUSCO scores are also very good at almost 99%.
I expected that this genome is ready to be used for downstream analyses and as such I fully endorse this manuscript in its current form.

Is the rationale for creating the dataset(s) clearly described? Yes
Are the protocols appropriate and is the work technically sound?Yes

Are the datasets clearly presented in a useable and accessible format? Yes
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: Insect genomics, bioinformatics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.This assembly provides valuable infomation for the genomics of Lepidoptera and for further phylognetic research in the evolutionary pattern in Lepidoptera.

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The methods for the sequencing and assembly used are appropriate and technically supported but It would be interesting to offer a calculation of the number of genes that encode proteins.
I suggest the addition of author names and year in the first mention of the species in the text ( Tethea ocularis, Tethea or, Diloba caeruleocephala, T. ocularis f. fusca and Tetheella fluctuosa).

Is the rationale for creating the dataset(s) clearly described? Yes
Are the protocols appropriate and is the work technically sound?Yes Are sufficient details of methods and materials provided to allow replication by others?Yes Are the datasets clearly presented in a useable and accessible format?Yes Competing Interests: No competing interests were disclosed.
Reviewer Expertise: Molecular genetics on Lepidoptera with special interest in evolutionary proccesses.
I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.
Figure of Eight Diloba caeruleocephala (Waring et al., 2017).The darker melanic form of Tethea ocularis f. fusca was first recorded in the south and east of England in the 1940's and has slowly spread across its UK range to become the dominant form (Skinner, 2009).The current UK range includes most of south-eastern Scotland, and all of England and Wales, with the exception of Cumberland (Waring et al., 2017), although it was first recorded in neighbouring Westmoreland in 1983 (Briggs, 1984).It is not present in Ireland but reached the Isle of Man in 2002 (Skinner, 2009).Tethea ocularis is a species of broad-leaved woodland, hedgerows, parks and gardens, where it is on the wing at night from May to July (Waring et al., 2017).The larvae feed on aspen Populus tremula and other poplars Populus spp., in a cocoon between the leaves, from mid-July to September.Tethea ocularis overwinters as a pupa (Waring et al., 2017).

Figure 2 .
Figure 2. Genome assembly of Tethea ocularis, ilTetOcul2.1:metrics.The BlobToolKit snail plot shows N50 metrics and BUSCO gene completeness.The main plot is divided into 1,000 size-ordered bins around the circumference with each bin representing 0.1% of the 339,142,399 bp assembly.The distribution of scaffold lengths is shown in dark grey with the plot radius scaled to the longest scaffold present in the assembly (15,953,683 bp, shown in red).Orange and pale-orange arcs show the N50 and N90 scaffold lengths (12,138,052 and 7,792,192 bp), respectively.The pale grey spiral shows the cumulative scaffold count on a log scale with white scale lines showing successive orders of magnitude.The blue and pale-blue area around the outside of the plot shows the distribution of GC, AT and N percentages in the same bins as the inner plot.A summary of complete, fragmented, duplicated and missing BUSCO genes in the lepidoptera_odb10 set is shown in the top right.An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/CAUVWG01/dataset/CAUVWG01/snail.

Figure 3 .
Figure 3. Genome assembly of Tethea ocularis, ilTetOcul2.1:BlobToolKit GC-coverage plot.Sequences are coloured by phylum.Circles are sized in proportion to sequence length.Histograms show the distribution of sequence length sum along each axis.An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/CAUVWG01/dataset/CAUVWG01/blob.

Figure 4 .
Figure 4. Genome assembly of Tethea ocularis, ilTetOcul2.1:BlobToolKit cumulative sequence plot.The grey line shows cumulative length for all sequences.Coloured lines show cumulative lengths of sequences assigned to each phylum using the buscogenes taxrule.An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/CAUVWG01/dataset/CAUVWG01/cumulative.

Figure 5 .
Figure 5. Genome assembly of Tethea ocularis, ilTetOcul2.1:Hi-C contact map of the ilTetOcul2.1 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=BwYrBgisQ3-PD8AWa8mIQA.

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Report 18 September 2024 https://doi.org/10.21956/wellcomeopenres.23763.r94816© 2024 Ortiz A. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Antonio S. Ortiz University of Murcia, Murcia, Spain Boyes & Wawman describes the genome assembly of Tethea ocularis and provides important genomic information about its size (near 340 Mb) in 31 chromosomes.A new genome sequenced from the moth family Drepanidae is added to others as Tethea albicostata (Bremer 1861) by Liang et al. (2023) published in Mitochondrial DNA part B, Drepana arcuata Walker, 1855 by Yadav et al. (2020) published in Data in Brief and ○ Drepana falcataria (Linnaeus, 1758) by Boyes & Lewis (2023), ○ Watsonalla binaria (Hufnagel, 1767) by Boyes & Chua (2023) and ○ Achlya flavicornis (Linnaeus, 1758) by Crowley & Phillips (2023), the last three published in Wellcome Open Research (WOR).

Table 1 :
Proposed standards and metrics for defining genome assembly quality" from Rhie et al. (2021).
to sample to eliminate shorter fragments and concentrate the DNA.The concentration of the sheared and purified DNA was assessed using a Nanodrop spectrophotometer and Qubit Fluorometer and Qubit dsDNA High Sensitivity Assay kit.Fragment size distribution was evaluated by running the sample on the FemtoPulse system.RNA Broad-Range Assay kit.Analysis of the integrity of the RNA was done using the Agilent RNA 6000 Pico Kit and Eukaryotic Total RNA assay.Protocols developed by the WSI Tree of Life laboratory are publicly available on protocols.io(Denton et al., 2023b).