An integrated strategy involving high‐throughput sequencing to characterize an unknown GM wheat event in Canada

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dc.contributor.author
Gagnon, Marie‐Claude
Duceppe, Marc‐Olivier
Colville, Adam
Pope, Louise
Côté, Marie‐José
Ogunremi, Dele
dc.date.accepted
2023-11-04
dc.date.accessioned
2024-08-29T15:18:52Z
dc.date.available
2024-08-29T15:18:52Z
dc.date.issued
2023-12-05
dc.date.submitted
2023-05-17
dc.description.abstract - en
Glyphosate-resistant wheat plants were discovered in southern Alberta in 2017, representing an unauthorized GM release in Canada. The Canadian Food Inspection Agency undertook a series of experiments to characterize and identify this unknown GM wheat, as well as to develop and validate construct-specific and event-specific qPCR assays. Results of PCR-based assays and Sanger sequencing indicated the presence of CaMV 35S promoter (p35S), Rice Actin 1 intron (RactInt1), CP4-EPSPS gene and nopaline synthase terminator (tNOS) elements in the unknown GM wheat. Genome walking and bead capture strategies, combined with high-throughput sequencing, were used to identify the 5′ and 3′ wheat junctions and the subsequent mapping of the insert to chromosome 3B of the wheat genome. A probable transformation vector, pMON25497, was recognized, and further testing identified the unknown GM wheat as MON71200 event, one of two events obtained with the pMON25497 vector. The two construct-specific assays targeted the junctions of the RactInt1 and the CP4-EPSPS elements and the CP4-EPSPS and tNOS elements, while the event-specific assay was located at the 3′ junction into the wheat genome. Both construct-specific and event-specific assays had limits of detection of 0.10% of MON71200 in a seed pool. As expected, the two construct-specific assays cross-reacted with other wheat and corn events containing the same elements in the same order. No cross-reactivity was observed for the event-specific assay. The integrated strategy employed in this study can serve as a model for other cases when facing similar challenges involving unknown GM events.
dc.identifier.citation
Gagnon, M., Duceppe, M., Colville, A., Pope, L., Côté, M., & Ogunremi, D. (2023). An integrated strategy involving high‐throughput sequencing to characterize an unknown GM wheat event in Canada. Plant Biotechnology Journal, 22(4), 904–914. https://doi.org/10.1111/pbi.14232
dc.identifier.doi
https://doi.org/10.1111/pbi.14232
dc.identifier.issn
1467-7652
dc.identifier.uri
https://science-ouverte.canada.ca/handle/123456789/2901
dc.language.iso
en
dc.publisher
John Wiley & Sons, Inc
dc.rights - en
Creative Commons Attribution 4.0 International (CC BY 4.0)
dc.rights - fr
Creative Commons Attribution 4.0 International (CC BY 4.0)
dc.rights.openaccesslevel - en
Gold
dc.rights.openaccesslevel - fr
Or
dc.rights.uri - en
https://creativecommons.org/licenses/by/4.0/
dc.rights.uri - fr
https://creativecommons.org/licenses/by/4.0/deed.fr
dc.subject - en
Agriculture
Science and technology
dc.subject - fr
Agriculture
Sciences et technologie
dc.subject.en - en
Agriculture
Science and technology
dc.subject.fr - fr
Agriculture
Sciences et technologie
dc.title - en
An integrated strategy involving high‐throughput sequencing to characterize an unknown GM wheat event in Canada
dc.type - en
Article
dc.type - fr
Article
local.article.journalissue
4
local.article.journaltitle
Plant Biotechnology Journal
local.article.journalvolume
22
local.pagination
904-914
local.peerreview - en
Yes
local.peerreview - fr
Oui
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