title: Genetics, structure, and prevalence of FP967 (CDC Triffid) T-DNA in flax.
abstract: The detection of T-DNA from a genetically modified flaxseed line (FP967, formally CDC Triffid) in a shipment of Canadian flaxseed exported to Europe resulted in a large decrease in the amount of flax planted in Canada. The Canadian flaxseed industry undertook major changes to ensure the removal of FP967 from the supply chain. This study aimed to resolve the genetics and structure of the FP967 transfer DNA (T-DNA). The FP967 T-DNA is thought to be inserted in at single genomic locus. The junction between the T-DNA and genomic DNA consisted of two inverted Right Borders with no Left Border (LB) flanking genomic DNA sequences recovered. This information was used to develop an event-specific quantitative PCR (qPCR) assay. This assay and an existing assay specific to the T-DNA construct were used to determine the genetics and prevalence of the FP967 T-DNA. These data supported the hypothesis that the T-DNA is present at a single location in the genome. The FP967 T-DNA is present at a low level (between 0.01 and 0.1%) in breeder seed lots from 2009 and 2010. None of the 11,000 and 16,000 lines selected for advancement through the Flax Breeding Program in 2010 and 2011, respectively, tested positive for the FP967 T-DNA, however. Most of the FP967 T-DNA sequence was resolved via PCR cloning and next generation sequencing. A 3,720 bp duplication of an internal portion of the T-DNA (including a Right Border) was discovered between the flanking genomic DNA and the LB. An event-specific assay, SAT2-LB, was developed for the junction between this repeat and the LB.
authors.affiliation: Department of Plant Sciences, 51 Campus Drive, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5A8 Canada.
authors.forename: Lester
authors.initials: L
authors.lastname: Young
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Joseph
authors.initials: J
authors.lastname: Hammerlindl
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Vivijan
authors.initials: V
authors.lastname: Babic
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Jamille
authors.initials: J
authors.lastname: McLeod
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Sharpe
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Chad
authors.initials: C
authors.lastname: Matsalla
authors.affiliation: National Research Council - Saskatoon, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9 Canada.
authors.forename: Faouzi
authors.initials: F
authors.lastname: Bekkaoui
authors.affiliation: Quantum BioSciences, 101 - 100 Research Drive, Saskatoon, Saskatchewan Canada.
authors.forename: Leigh
authors.initials: L
authors.lastname: Marquess
authors.affiliation: Department of Plant Sciences, 51 Campus Drive, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5A8 Canada.
authors.forename: Helen M
authors.initials: HM
authors.lastname: Booker
keywords: CDC Triffid; FP967; Flaxseed; Linum usitatissimum; T-DNA