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. 2019 Oct 17;9:14911. doi: 10.1038/s41598-019-51212-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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(A) Phylogenetic tree constructed with Maximum Likelihood of Brassica FLC from three diploid (B. rapa, B. nigra, B. oleracea) and one polyploid (B. napus) species with Ath.FLC as outgroup. cDNA sequences were extracted from the respective reference genomes, the sequence from A. thaliana was retrieved from TAIR. Sequence alignment was performed using CLUSTAL multiple sequence alignment by MUSCLE with Default parameters. Based on this alignment, a Maximum Likelihood tree was constructed using MEGA version 10.0.5. (B) Full gene structure including UTR (dark grey boxes), introns (light grey boxes) and exons (colored boxes). Black boxes represent extra exons. Box length is proportional to length in bp, the legend shows the length of 100 bp. The solid line in black shows the localization of COOLAIR, the dotted line in grey represents the approximate position of COLDAIR. (C) Exon structure including the position of sequence variants within the 5 accessions used for the initial RT-qPCR. Dark grey boxes represent UTRs, colored boxes represent exons in the same color code than for B, black boxes represent extra exons. Black lines represent a variant, black lines with a black triangle represent a non-synonymous SNP, annotated with the respective amino acid mutation.