Supplementary information of Towards carotenoid biofortification in wheat: identification of XAT-7A1, a multicopy tandem gene responsible for carotenoid esterification in durum wheat

[Description of methods used for collection/generation of data] 12870_2023_4431_MOESM1_ESM.xlsx. Orthologue/homoeologue sequences obtained from EnsemblPlants after BLASTn. 12870_2023_4431_MOESM2_ESM.xlsx. Genomic sequences of genes TraesCS7D02G094000, TraesCS4A02G397900, TRITD4Av1G231840 and TRITD4Av1G231510 were retrieved from Ensembl Plants and aligned using the multiple sequence alignment tool ClustalW. Primers were designed in the conserved 5’ and 3’ regions by using the NCBI Primer-Blast tool. SNP markers were designed following a Tetra-Primer ARMS (amplification refractory mutation system) strategy for SNPs detection. Primer1 web service (http://primer1.soton.ac.uk/primer1.html) was used for primer design. 12870_2023_4431_MOESM4_ESM.xlsx. Sequence editing, alignment and assembly were performed with SeqMan Pro Lasergene Software v17 (DNAStar, WI, US). The identity of the clones as GDSL esterase-lipase- like sequences was confirmed by BLASTn at NCBI. The coding sequences were predicted based on the exon-intron structure of TraesCS7D02G094000, and open reading frames were searched by using ORFfinder at NCBI (https://www.ncbi.nlm.nih.gov/orffinder). Signal peptide and cellular location of the expected proteins were predicted by SignalP 6.0 software 12870_2023_4431_MOESM5_ESM.xlsx. The genetic map was constructed using Joinmap(R) 5.0 [Kyazma(R), The Netherlands]. 12870_2023_4431_MOESM6_ESM.xlsx. Carotenoid content and profile was determined according to Rodríguez-Suárez C, Requena-Ramírez MD, Hornero-Méndez D, Atienza SG (2022). Chapter 4. The breeder’s tool-box for enhancing the content of esterified carotenoids in wheat: From extraction and profiling of carotenoids to marker-assisted selection of candidate genes. En: Carotenoids: Carotenoid and apocarotenoid biosynthesis, metabolic engineering and synthetic biology. pp. 99-125. Editor. Eleanore T. Wurtzel. Book series: Methods in Enzymology. Editorial: Academic Press. Elsevier. Hardcover ISBN: 9780323913539.

This research was financed by project PID2021-122152NB-I00 funded by MCIN/AEI/10.13039/501100011033/ and by ERDF “ERDF A way of making Europe”. M.D.R.-R. was supported by PRE2018-084037 funded by MCIN/AEI/10.13039/501100011033 and ESF “ESF investing in your future”. DH-M is member of the Spanish Carotenoid Network (CaRed), grant RED2022-134577-T. SA, CR-S and MDR-R are members of CeReS Network, grant RED2022-134922-T. Both networks are funded by MCIN/AEI/10.13039/501100011033.

12870_2023_4431_MOESM1_ESM.xlsx 12870_2023_4431_MOESM2_ESM.xlsx 12870_2023_4431_MOESM3_ESM.xlsx 12870_2023_4431_MOESM4_ESM.xlsx 12870_2023_4431_MOESM5_ESM.xlsx 12870_2023_4431_MOESM6_ESM.xlsx 12870_2023_4431_MOESM1_ESM.xlsx contains the results of the Search for XAT candidate genes in durum wheat genomes using XAT-7D (TraesCS7D02G094000) from common wheat as gene model. 12870_2023_4431_MOESM2_ESM.xlsx contains the list of primers designed in this work. 12870_2023_4431_MOESM3_ESM.xlsx contains the list of landraces showing positive amplification for XAT candidate gene. 12870_2023_4431_MOESM4_ESM.pdf contains the alignment of XAT-7A1 and XAT-7D proteins. The predicted active sites are shown with black triangles. Signal peptide in positions 1–22 is highlighted. 12870_2023_4431_MOESM5_ESM.xlsx contains the genetic map for the F2 population derived from the cross BGE047535 × 'Athoris' using DArTSeq markers. 12870_2023_4431_MOESM6_ESM.xlsx contains the carotenoid content and profile of the F2 population derived from the cross BGE047535 x Athoris.

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