The genome sequence of the setaceous Hebrew character, Xestia c-nigrum, (Linnaeus, 1758)

We present a genome assembly from an individual male Xestia c-nigrum (the setaceous Hebrew character; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence is 760 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 15.3 kilobases in length.


Background
Known to most British lepidopterists as setaceous Hebrew character, Xestia c-nigrum is generally referred to as the spotted cutworm in the pest control literature. The latter name is derived from the appearance of the caterpillar while 'c-nigrum' and 'Hebrew character' reference the distinctive black marking on the forewing.
X. c-nigrum is a familiar, widespread species across Asia, Europe and North America. In much of its range, including Britain, there are two generations in a year, the second usually much larger. The summer and autumn cohort might be larger due to increased survival (larvae of the spring cohort over-winter) and/or immigration from further south (Clancy et al., 2012). Larvae feed on a variety of herbaceous plants and over-winter as diapausing larvae, growing at a slower rate than non-diapausing larvae (Honek, 1979). A granulovirus isolated from X. c-nigrum has been used to develop virus-based biopesticides against agricultural pest moths, e.g. (Goto et al., 2015). This is the second whole genome sequence for a Xestia moth species, following that of X. xanthographa (Boyes & Holland, 2022). The sequenced individual was collected at Hever Castle during a collecting trip for the Natural History Museum Darwin Tree of Life sampling team.

Genome sequence report
The genome was sequenced from one male X. c-nigrum ( Figure 1) collected from Hever Castle, England, UK (latitude 51.188, longitude 0.12). A total of 35-fold coverage in Pacific Biosciences single-molecule HiFi long reads and 58-fold coverage in 10X Genomics read clouds were generated. Primary assembly contigs were scaffolded with chromosome conformation Hi-C data. Manual assembly curation corrected nine (9) missing/misjoins, reducing the assembly length by 0.03% and the scaffold number by 14%, and increasing the scaffold N50 by 0.1%.
The final assembly has a total length of 760 Mb in 43 sequence scaffolds with a scaffold N50 of 25.7 Mb (Table 1). Most of the assembly sequence (99.9%) was assigned to 31 chromosomal-level scaffolds confirmed by Hi-C data, representing 30 autosomes named in order of size and the Z chromosome    Table 2). The assembly has a BUSCO 5.3. 2 (Manni et al., 2021) completeness of 98.8% using the lepidoptera_odb10 reference set.
While not fully phased, the assembly deposited is of one haplotype. Contigs corresponding to the second haplotype have also been deposited.

Sample acquisition and nucleic acid extraction
A male X. c-nigrum (ilXesCnig1) was collected using a light trap and identified by Gavin Broad (Natural History Museum) from Hever Castle, United Kingdom (latitude 51.188, longitude 0.12). The sample was preserved on dry ice by Laura Sivess. A second X. c nigrum (ilXesCnig2) was collected using a light trap and identified by Douglas Boyes (Natural History Museum) from Wytham Woods, United Kingdom (latitude 51.772, longitude -1.338).
DNA was extracted from head and thorax tissue of ilXesCnig1 at the Wellcome Sanger Institute (WSI) Scientific Operations core using the Qiagen MagAttract HMW DNA kit, according to the manufacturer's instructions. RNA was extracted from abdomen tissue of ilXesCnig2 in the Tree of Life   Biosciences SEQUEL II (HiFi), Illumina HiSeq 4000 (RNA-Seq) and Illumina NovaSeq 6000 (10X) instruments. Hi-C data were also generated from head tissue of ilXesCnig1 using the Arima v2 kit and sequenced on the Illumina NovaSeq 6000 instrument.

Robert Simpson
The New Zealand Institute of Plant and Food Research Limited, Palmerston North, New Zealand This manuscript is a data note, briefly describing the assembly of the genome for the Noctuid setaceous Hebrew character. As a note it is necessarily brief, but it is sufficient to follow the methods used in sequencing and assembly and show the completeness of the submitted genome. Xestia c-nigrum is a cosmopolitan agricultural pest, continuing to extend its range and also a generalist feeder. This genome assembly will be an important tool both for those working directly on the insect, but also for those interested in generalist feeder and invasive species. There are no major problems with the note, but the following minor alterations should be considered.
There is more information about the mitochondrion in the abstract than in the body of the note. It would be helpful to add a short sentence stating exact size, and whether it is complete. 1.
The reviewer understands the authors' desire to include all material, but the second haplotype data is not well integrated with the remainder of the note. Either delete this or add more information. What amount of coverage? How much identity with the first haplotype? Any observed chromosome rearrangements? 2.
If the second haplotype is included, the sentence beginning 'Contigs corresponding to the second…' should be amended to 'Contigs corresponding to a second…'.

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?