(A) Sampling locations and frequencies of yellow and white males in Finland, Scotland, and Estonia. (B) Males of the white and yellow colour morphs (credit: Samuel Waldron).
(A) Quantitative trait locus (QTL) analysis of white and yellow F1 males (n = 172) reveals a 500 kb region significant on scaffold 206 of the yellow reference, part of linkage group 9. The dotted …
MQ is lower within the duplicated region (between the dotted lines): average MQ for the duplication including all samples is 36.52, whereas for the rest of the scaffold, not including the …
Top: Yellow alleles have the larger 163 bp band, while white alleles have a 35 bp deletion producing the smaller ~128 bp band. Heterozygotes have a copy of each. Bottom: A 449 bp region within the …
Unedited gel image showing the genotyping primers.
Yellow-e sequences from D. melanogaster and H. melpomene are also included. Values are substitutions per site.
(A) In pink are genes that are significantly differentially expressed between yellow and white morphs at the pre-melanin stage. Valkea is the most differentially expressed (DE) gene (i.e. gene with …
Multidimensional scaling of gene expression shows that samples cluster by developmental stage rather than male colour morph. Developmental stage names have been abbreviated: 72h = 72 hours, 5d = 5 …
Wildtype WW and yy morphs (top), and the dorsal and ventral sides of one of the CRISPR knockout males (bottom).
Variation in melanin amount is likely due to the lineage, with (A) coming from the Estonian line, and (B–D) coming from the Finnish line. (E) Female showing a mosaic knockout phenotype with left …
The brighter pink colour in the UV photos (right) shows the areas with highest UV reflectance. UV reflectance is lost in the CRISPR knockout males.
The top row of each block shows the reference sequence, and the rows underneath are the mutant individuals. Pink sequences denote the guide RNA sequence. Four different guides produced mutants (one …
The coloured section of the wings were measured on male forewings (A), male hindwings (B), female forewings (C), and female hindwings (D). (E) shows measurements for each wing of the one CRISPR …
Measurements for yellow, white, and black portions of the hindwing, plus the standard (Std) are shown.
White (A) and yellow (B) A. plantaginis morphs are compared to ommochrome-containing Heliconius wings (C) and a Xanthurenic acid standard (D).
The black portions of the wings show the presence of dopamine-derived eumelanin components pyrrole-2,3-dicarboxylic acid (PDCA) and pyrrole-2,3,5-tricarboxylic acid (PTCA). Phthalic acid is the …
Supplementary tables.
(A) 21 genes are within in the QTL interval. Start and end positions shown are on scaffold 206 in the yellow A. plantaginis reference genome. Gene sequences were blasted against Heliconius melpomene and searched for in FlyBase. Apla gene names from annotations produced by Yen et al., 2020. (B) Primers used for genotyping. Tested using GoTaq Flexi buffer and GoTaq DNA polymerase, with annealing temperature of 57°C for 35 cycles. ‘Ye12’ primers surround a small deletion in white alleles, producing a 163 bp product from Y alleles and 128 bp product from W alleles. ‘Dup5’ primers amplify a 449 bp sequence within the duplicated sequence only in moths with at least one W allele. See Figure 2—figure supplement 4 for gel images. (C) List of differentially expressed genes found in the linkage group containing scaffold 419 (WW reference). (D) Details of the number of eggs injected with each sgRNA and those which produced adult moths. (E) Sample list of all lab cross individuals used in linkage mapping. (F) Sample list of all wild samples used. (G) CRISPR sgRNAs sequences. (H) Elution gradient used in pheomelanin HPLC analysis. (I). Waveform of disposable working electrode in pheomelanin analysis.