Abstract
We present a genome assembly from an individual female Maniola jurtina (the meadow brown; Arthropoda; Insecta; Lepidoptera; Nymphalidae). The genome sequence is 402 megabases in span. The complete assembly is scaffolded into 30 chromosomal pseudomolecules, with the W and Z sex chromosome assembled. Gene annotation of this assembly on Ensembl has identified 12,502 protein coding genes.
Keywords: Maniola jurtina, meadow brown, genome sequence, chromosomal
Species taxonomy
Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; Neoptera; Endopterygota; Lepidoptera; Glossata; Ditrysia; Papilionoidea; Nymphalidae; Satyrinae; Satyrini; Maniolina; Maniola; Maniola jurtina (Linnaeus, 1758) (NCBI:txid191418).
Introduction
The meadow brown Maniola jurtina is a common, Palearctic butterfly occurring throughout Europe, the Middle East, and North Africa ( gbif.org, 2021). Both widespread and often abundant, the species is associated with almost any grassy habitats ( South, 1906), reaching highest densities in areas where grazing or other pressures keep the sward at an intermediate height ( Maitland Emmet & Heath, 1989). Although M. jurtina is consistently univoltine, emergence occurs over a prolonged period in summer, which varies in length geographically and with habitat types ( Brakefield, 1987). In many Mediterranean populations, females aestivate during the hottest months of the year ( Scali, 1971). Eggs are laid singly or in small clusters, both on individual blades of grass or loose into a suitable tuft ( Maitland Emmet & Heath, 1989), preferentially on Poa, Agrostis and Lolium. Larvae overwinter, but do not undergo true diapause, and feed intermittently in warm spells. Pupae show considerable variation in colouration which is affected by light and temperature ( Brakefield, 1979). The species exhibits a great deal of phenotypic variation both within and between populations ( Thomson, 1969). Four sub-species are known from the British Isles – ssp. splendida, insularis, iernes, and cassiteridum – although the validity of these taxa is questionable ( Weir & Others, 2016), since they seem to be phenotypic extremes at opposing ends of clines ( Maitland Emmet & Heath, 1989). In their pioneering work in ecological genetics, Ford and Dowdeswell considered the evolutionary factors shaping variation in the spot patterning of the underside of the hindwings in M. jurtina, initially on the Isles of Scilly, then the British mainland (reviewed in ( Ford, 1964) and ( Dowdeswell, 1981). Several early studies ( Bigger, 1960; Federley, 1938; Lorković, 1941), summarised in ( Robinson, 1971), report a karyotype of 29 chromosomes. The genome size has been estimated as 367.3 Mb ( Mackintosh et al., 2019). We note the publication of a de novo genome assembly of M. jurtina by ( Singh et al., 2020) and believe that the sequence described here, generated as part of the Darwin Tree of Life project, will further aid understanding of the biology of this butterfly.
Genome sequence report
The genome was sequenced from a female M. jurtina (ilManJurt1; Figure 1A, B) collected from Aberlady Bay, East Lothian, Scotland, UK (latitude 56.019964, longitude -2.85808). Hi-C data were generated from another individual (ilManJurt3; Figure 1E, F) collected from East Linton, East Lothian, Scotland, UK (latitude 55.977161, longitude -2.667545). A total of 76-fold coverage in Pacific Biosciences single-molecule long reads (N50 14 kb) and 88-fold coverage in 10X Genomics read clouds were generated. Primary assembly contigs were scaffolded with chromosome conformation Hi-C data. Manual assembly curation corrected 24 missing/misjoins and removed two haplotypic duplications, reducing the assembly size by 1.67% and scaffold number by 24.39%.
Figure 1. Fore and hind wings of Maniola jurtina specimens from which the genome was sequenced.
( A) Dorsal surface view of wings from specimen SC_MJ_1353 (ilManJurt1) from Aberlady, Scotland, UK, used to generate Pacific Biosciences and 10X data. ( B) Ventral surface view of wings from specimen SC_MJ_1353 (ilManJurt1) from Aberlady, Scotland, UK, used to generate Pacific Biosciences and 10X data. ( C) Dorsal surface view of wings from specimen SC_MJ_1360 (ilManJurt2) from East Linton, Scotland, UK, used to generate RNA-Seq data. ( D) Ventral surface view of wings from specimen SC_MJ_1360 (ilManJurt2) from East Linton, Scotland, UK, used to generate RNA-Seq data. ( E) Dorsal surface view of wings from specimen SC_MJ_1361 (ilManJurt3) from East Linton, Scotland, UK, used to generate Hi-C data. ( F) Ventral surface view of wings from specimen SC_MJ_1361 (ilManJurt3) from East Linton, Scotland, UK, used to generate Hi-C data.
The final assembly has a total length of 402 Mb in 30 sequence scaffolds with a scaffold N50 of 15 Mb ( Table 1). Of the assembly sequence, 100% was assigned to 30 chromosomal-level scaffolds, representing 28 autosomes (numbered by sequence length), and the W and Z sex chromosome ( Figure 2– Figure 5; Table 2). The assembly has a BUSCO ( Simão et al., 2015) completeness of 98.3% using the lepidoptera_odb9 reference set. While not fully phased, the assembly deposited is of one haplotype. Contigs corresponding to the second haplotype have also been deposited.
Figure 2. Genome assembly of Maniola jurtina, ilManJurt1.1: metrics.
The BlobToolKit Snailplot 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 402,054,102 bp assembly. The distribution of chromosome lengths is shown in dark grey with the plot radius scaled to the longest chromosome present in the assembly (17,318,487 bp, shown in red). Orange and pale-orange arcs show the N50 and N90 chromosome lengths (15,090,120 and 9,430,113 bp), respectively. The pale grey spiral shows the cumulative chromosome 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/ilManJurt1.1/dataset/CAJOSP01/snail.
Figure 5. Genome assembly of Maniola jurtina, ilManJurt1.1: Hi-C contact map.
Hi-C contact map of the ilManJurt1.1 assembly, visualised in HiGlass. Chromosomes are shown in size order from left to right and top to bottom.
Table 1. Genome data for Maniola jurtina, ilManJurt1.1.
| Project accession data | |
|---|---|
| Assembly identifier | ilManJurt1.1 |
| Species | Maniola jurtina |
| Specimen | ilManJurt1 (genome assembly); ilManJurt2, ilManJurt5 (RNA-Seq); ilManJurt3 (Hi-C) |
| NCBI taxonomy ID | NCBI:txid191418 |
| BioProject | PRJEB43535 |
| BioSample ID | SAMEA7523158 |
| Isolate information | Female, whole organisms |
| Raw data accessions | |
| PacificBiosciences SEQUEL II | ERR6576323 |
| 10X Genomics Illumina | ERR6054518–ERR6054521 |
| Hi-C Illumina | ERR6054522 |
| Illumina PolyA RNAseq | ERR6054523, ERR6787422 |
| Genome assembly | |
| Assembly accession | GCA_905333055.1 |
| Accession of alternate haplotype | GCA_905333105.1 |
| Span (Mb) | 402 |
| Number of contigs | 53 |
| Contig N50 length (Mb) | 13 |
| Number of scaffolds | 32 |
| Scaffold N50 length (Mb) | 15 |
| Longest scaffold (Mb) | 17 |
| BUSCO * genome score | C:98.3%[S:97.7%,D:0.6%],F:0.3%,M:1.4%,n:5286 |
*BUSCO scores based on the lepidoptera_odb10 BUSCO set using v5.1.2. C= complete [S= single copy, D=duplicated], F=fragmented, M=missing, n=number of orthologues in comparison. A full set of BUSCO scores is available at https://blobtoolkit.genomehubs.org/view/ilManJurt1.1/dataset/CAJOSP01/busco.
Figure 3. Genome assembly of Maniola jurtina, ilManJurt1.1: GC coverage.
BlobToolKit GC-coverage plot. Chromosomes are coloured by phylum. Circles are sized in proportion to chromosome length. Histograms show the distribution of scaffold length sum along each axis. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/ilManJurt1.1/dataset/CAJOSP01/blob.
Figure 4. Genome assembly of Maniola jurtina, ilManJurt1.1: cumulative sequence.
BlobToolKit cumulative sequence plot. The grey line shows cumulative length for all chromosomes. Coloured lines show cumulative lengths of chromosomes assigned to each phylum using the buscogenes taxrule. An interactive version of this figure is available at https://blobtoolkit.genomehubs.org/view/ilManJurt1.1/dataset/CAJOSP01/cumulative.
Gene annotation
The Ensembl gene annotation system ( Aken et al., 2016) was used to generate annotation for the Maniola jurtina assembly (GCA_905333055.1, see https://rapid.ensembl.org/Maniola_jurtina_GCA_905333055.1/; Table 1). The 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) and OrthoDB ( Kriventseva et al., 2008). Prediction tools, CPC2 ( Kang et al., 2017) and RNAsamba ( Camargo et al., 2020), were used to aid determination of protein coding genes.
Table 2. Chromosomal pseudomolecules in the genome assembly of Maniola jurtina, ilManJurt1.1.
| INSDC accession | Chromosome | Size (Mb) | GC% |
|---|---|---|---|
| HG995207.1 | 1 | 17.32 | 36.8 |
| HG995209.1 | 2 | 17.19 | 36.6 |
| HG995210.1 | 3 | 16.76 | 36.8 |
| HG995211.1 | 4 | 16.67 | 36.8 |
| HG995212.1 | 5 | 16.23 | 36.7 |
| HG995213.1 | 6 | 16.20 | 36.8 |
| HG995214.1 | 7 | 15.71 | 36.3 |
| HG995215.1 | 8 | 15.42 | 36.9 |
| HG995216.1 | 9 | 15.23 | 37 |
| HG995217.1 | 10 | 15.17 | 36.6 |
| HG995218.1 | 11 | 15.11 | 36.8 |
| HG995219.1 | 12 | 15.09 | 36.7 |
| HG995220.1 | 13 | 14.83 | 36.9 |
| HG995221.1 | 14 | 14.70 | 36.9 |
| HG995222.1 | 15 | 14.15 | 36.9 |
| HG995223.1 | 16 | 14.03 | 36.8 |
| HG995224.1 | 17 | 13.72 | 36.9 |
| HG995225.1 | 18 | 13.54 | 36.9 |
| HG995226.1 | 19 | 13.29 | 37.1 |
| HG995227.1 | 20 | 13.28 | 37 |
| HG995228.1 | 21 | 12.26 | 37.3 |
| HG995229.1 | 22 | 12.13 | 36.9 |
| HG995230.1 | 23 | 10.05 | 37.2 |
| HG995231.1 | 24 | 9.43 | 37.4 |
| HG995232.1 | 25 | 8.57 | 37.3 |
| HG995233.1 | 26 | 7.75 | 38.7 |
| HG995235.1 | 27 | 6.95 | 37.7 |
| HG995236.1 | 28 | 6.66 | 38.1 |
| HG995234.1 | W | 7.35 | 38.3 |
| HG995208.1 | Z | 17.21 | 36.9 |
| HG995237.1 | MT | 0.02 | 20.3 |
| - | Unplaced | 0.04 | 37.4 |
Methods
Sample acquisition and nucleic acid extraction
Four female M. jurtina samples (genome assembly, ilManJurt1; RNAseq, ilManJurt2, ilManJurt5; Hi-C, ilManJurt3) were collected and used for sequencing. Sample ilManJurt1 was caught in Aberlady, East Lothian, UK (latitude 56.019964, longitude -2.85808). Samples ilManJurt2 ( Figure 1C, D), ilManJurt3 and ilManJurt5 were caught in East Linton, East Lothian, UK (latitude 56.019964, longitude -2.85808). All samples were collected using a handnet by Konrad Lohse, University of Edinburgh, and snap-frozen in liquid nitrogen.
DNA was extracted at the Wellcome Sanger Institute (WSI) Scientific Operations core from the whole organism using the Qiagen MagAttract HMW DNA kit, according to the manufacturer’s instructions. RNA was extracted in the Tree of Life Laboratory at the WSI using TRIzol (Invitrogen), according to the manufacturer’s instructions. RNA was then eluted in 50 μl RNAse-free water and its concentration assessed using a Nanodrop spectrophotometer and Qubit Fluorometer using the Qubit RNA Broad-Range (BR) Assay kit. Analysis of the integrity of the RNA was done using Agilent RNA 6000 Pico Kit and Eukaryotic Total RNA assay.
Sequencing
Pacific Biosciences HiFi circular consensus and 10X Genomics Chromium read cloud sequencing libraries were constructed according to the manufacturers’ instructions. Poly(A) RNA-Seq libraries were constructed using the NEB Ultra II RNA Library Prep kit. Sequencing was performed by the Scientific Operations core at the Wellcome Sanger Institute on Pacific Biosciences SEQUEL II (HiFi), Illumina HiSeq X (10X) and Illumina HiSeq 4000 (RNA-Seq) instruments. Hi-C data were generated using the Qiagen EpiTect Hi-C kit and sequenced on HiSeq X.
Genome assembly
Assembly was carried out with Hifiasm ( Cheng et al., 2021). Haplotypic duplication was identified and removed with purge_dups ( Guan et al., 2020). One round of polishing was performed by aligning 10X Genomics read data to the assembly with longranger align, calling variants with freebayes ( Garrison & Marth, 2012). The assembly was then scaffolded with Hi-C data ( Rao et al., 2014) using SALSA2 ( Ghurye et al., 2019). The assembly was checked for contamination and corrected using the gEVAL system ( Chow et al., 2016) as described previously ( Howe et al., 2021). Manual curation ( Howe et al., 2021) was performed using gEVAL, HiGlass ( Kerpedjiev et al., 2018) and Pretext. The mitochondrial genome was assembled using MitoHiFi ( Uliano-Silva et al., 2021). The genome was analysed and BUSCO scores generated within the BlobToolKit environment ( Challis et al., 2020). Table 3 contains a list of all software tool versions used, where appropriate.
Table 3. Software tools used.
| Software tool | Version | Source |
|---|---|---|
| Hifiasm | 0.12 | Cheng et al., 2021 |
| purge_dups | 1.2.3 | Guan et al., 2020 |
| SALSA2 | 2.2 | Ghurye et al., 2019 |
| longranger align | 2.2.2 | https://support.10xgenomics.com/genome-exome/software/pipelines/latest/advanced/other-pipelines |
| freebayes | v1.3.1-17-gaa2ace8 | Garrison & Marth, 2012 |
| MitoHiFi | 1.0 | https://github.com/marcelauliano/MitoHiFi |
| gEVAL | N/A | Chow et al., 2016 |
| HiGlass | 1.11.6 | Kerpedjiev et al., 2018 |
| PretextView | 0.1.x | https://github.com/wtsi-hpag/PretextView |
| BlobToolKit | 2.6.2 | Challis et al., 2020 |
Ethical/compliance issues
The materials that have contributed to this genome note were supplied by a Tree of Life collaborator. 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 undertaken according to a Research Collaboration Agreement or Material Transfer Agreement entered into by the Tree of Life collaborator, Genome Research Limited (operating as the Wellcome Sanger Institute) and in some circumstances other Tree of Life collaborators.
Data availability
European Nucleotide Archive: Maniola jurtina (meadow brown). Accession number PRJEB43535; https://identifiers.org/ena.embl/PRJEB43535.
The genome sequence is released openly for reuse. The M. jurtina genome sequencing initiative is part of the Darwin Tree of Life (DToL) project. All raw sequence data and the assembly have been deposited in INSDC databases. Raw data and assembly accession identifiers are reported in Table 1.
Funding Statement
This work was supported by Wellcome through core funding to the Wellcome Sanger Institute (206194) and the Darwin Tree of Life Discretionary Award (218328). KL is supported by a NERC fellowship (NE/L011522/1) and an ERC grant (ModelGenom Land 757648). JW is supported by an EastBio PhD studentship.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
[version 1; peer review: 2 approved, 1 approved with reservations]
Author information
Members of the Darwin Tree of Life Barcoding collective are listed here: https://doi.org/10.5281/zenodo.4893704.
Members of the Wellcome Sanger Institute Tree of Life programme collective are listed here: https://doi.org/10.5281/zenodo.5377053.
Members of Wellcome Sanger Institute Scientific Operations: DNA Pipelines collective are listed here: https://doi.org/10.5281/zenodo.4790456.
Members of the Tree of Life Core Informatics collective are listed here: https://doi.org/10.5281/zenodo.5013542.
Members of the Darwin Tree of Life Consortium are listed here: https://doi.org/10.5281/zenodo.4783559.
References
- Aken BL, Ayling S, Barrell D, et al. : The Ensembl Gene Annotation System. Database (Oxford). 2016;baw093. 10.1093/database/baw093 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bigger TRL: Chromosome Numbers of Lepidoptera. Part I. Entomologist’s Gazette. 1960;11:149–52. [Google Scholar]
- Brakefield PM: An Experimental Study of the Maintenance of Variation in Spot Pattern in Maniola Jurtina. Thesis Ph.D., Liverpool University.1979. [Google Scholar]
- Brakefield PM: Geographical Variability In, and Temperature Effects On, the Phenology of Maniola Jurtina and Pyronia Tithonus (Lepidoptera, Satyrinae) in England and Wales. Ecol Entomol. 1987;12(2):139–48. Reference Source [Google Scholar]
- Camargo AP, Sourkov V, Pereira GAG, et al. : RNAsamba: Neural Network-Based Assessment of the Protein-Coding Potential of RNA Sequences. NAR Genom Bioinform. 2020;2(1):lqz024. 10.1093/nargab/lqz024 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Challis R, Richards E, Rajan J, et al. : BlobToolKit - Interactive Quality Assessment of Genome Assemblies. G3 (Bethesda). 2020;10(4):1361–74. 10.1534/g3.119.400908 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheng H, Concepcion GT, Feng X, et al. : Haplotype-Resolved de Novo Assembly Using Phased Assembly Graphs with Hifiasm. Nat Methods. 2021;18(2):170–75. 10.1038/s41592-020-01056-5 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chow W, Brugger K, Caccamo M, et al. : gEVAL - a web-based browser for evaluating genome assemblies. Bioinformatics. 2016;32(16):2508–10. 10.1093/bioinformatics/btw159 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dowdeswell WH: The Life of the Meadow Brown. Heinemann Educational Books.1981. Reference Source [Google Scholar]
- Federley H: Chromosomenzahlen Finnlän-Discher Lepidopteren. Hereditas. 1938;24(4):397–464. 10.1111/j.1601-5223.1938.tb03219.x [DOI] [Google Scholar]
- Ford EB: Ecological Genetics. Metheun.1964. Reference Source [Google Scholar]
- Garrison E, Marth G: Haplotype-Based Variant Detection from Short-Read Sequencing. arXiv: 1207.3907.2012. Reference Source [Google Scholar]
- gbif.org, Registry-Migration: GBIF Backbone Taxonomy.GBIF Secretariat.2021. 10.15468/39OMEI [DOI] [Google Scholar]
- Ghurye J, Rhie A, Walenz BP, et al. : Integrating Hi-C Links with Assembly Graphs for Chromosome-Scale Assembly. PLoS Comput Biol. 2019;15(8):e1007273. 10.1371/journal.pcbi.1007273 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guan D, McCarthy SA, Wood J, et al. : Identifying and removing haplotypic duplication in primary genome assemblies. Bioinformatics. 2020;36(9):2896–2898. 10.1093/bioinformatics/btaa025 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howe K, Chow W, Collins J, et al. : Significantly Improving the Quality of Genome Assemblies through Curation. GigaScience. 2021;10(1):giaa153. 10.1093/gigascience/giaa153 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kang YJ, Yang DC, Kong L, et al. : CPC2: A Fast and Accurate Coding Potential Calculator Based on Sequence Intrinsic Features. Nucleic Acids Res. 2017;45(W1):W12–16. 10.1093/nar/gkx428 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kerpedjiev P, Abdennur N, Lekschas F, et al. : HiGlass: Web-Based Visual Exploration and Analysis of Genome Interaction Maps. Genome Biol. 2018;19(1):125. 10.1186/s13059-018-1486-1 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kriventseva EV, Rahman N, Espinosa O, et al. : OrthoDB: The Hierarchical Catalog of Eukaryotic Orthologs. Nucleic Acids Res. 2008;36(Database issue):D271–75. 10.1093/nar/gkm845 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lorković Z: Die Chromosomenzahlen in der Spermatogenese der Tagfalter. Chromosoma. 1941;2(1):155–91. 10.1007/BF00325958 [DOI] [Google Scholar]
- Mackintosh A, Laetsch DR, Hayward A, et al. : The Determinants of Genetic Diversity in Butterflies. Nat Commun. 2019;10(1):3466. 10.1038/s41467-019-11308-4 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maitland Emmet AM, Heath J: The Moths and Butterflies of Great Britain and Ireland: Hesperiidae - Nymphalidae, the Butterflies.Harley Books,1989. Reference Source [Google Scholar]
- Rao SSP, Huntley MH, Durand NC, et al. : A 3D Map of the Human Genome at Kilobase Resolution Reveals Principles of Chromatin Looping. Cell. 2014;159(7):1665–80. 10.1016/j.cell.2014.11.021 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robinson R: Lepidoptera Genetics.Pergamon Press,1971. 10.1016/C2013-0-01588-5 [DOI] [Google Scholar]
- Scali V: SPOT DISTRIBUTION IN MANIOLA JURTINA (L.) (LEPIDOPTERA SATYRIDAE): TUSCAN MAINLAND 1967--1969. Monitore Zoologico Italiano-Italian Journal of Zoology. 1971;5(3):147–63. Reference Source [Google Scholar]
- Simão FA, Waterhouse RM, Ioannidis P, et al. : BUSCO: Assessing Genome Assembly and Annotation Completeness with Single-Copy Orthologs. Bioinformatics. 2015;31(19):3210–12. 10.1093/bioinformatics/btv351 [DOI] [PubMed] [Google Scholar]
- Singh KS, Hosken DJ, Wedell N, et al. : De Novo Genome Assembly of the Meadow Brown Butterfly, Maniola Jurtina. G3 (Bethesda). 2020;10(5):1477–84. 10.1534/g3.120.401071 [DOI] [PMC free article] [PubMed] [Google Scholar]
- South R: The Butterflies of the British Isles.Frederick Warne & Co,1906. Reference Source [Google Scholar]
- Thomson G: Maniola (Epinephile) Jurtina (L.)(Lep. Satyridae) and Its Forms. Entomol Rec J Variation. 1969. Reference Source [Google Scholar]
- Uliano-Silva M, Nunes JGF, Krasheninnikova K, et al. : marcelauliano/MitoHiFi: mitohifi_v2.0.2021. 10.5281/zenodo.5205678 [DOI] [Google Scholar]
- UniProt Consortium: UniProt: A Worldwide Hub of Protein Knowledge. Nucleic Acids Res. 2019;47(D1):D506–15. 10.1093/nar/gky1049 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weir JC and Others: Intraspecific Taxonomy in the Lepidoptera. Br J Ent Nat Hist. 2016;29(3):144–54. Reference Source [Google Scholar]