Agrobacterium mediated transfer of chlorsulfuron resistance to commercial flax cultivars

Abstract

Agrobacterium tumefaciens carrying a disarmed Ti-plasmid vector containing a chimeric NPT-ll gene and a mutant acetolactate synthase gene (conferring resistance to the herbicide chlorsulfuron) from Arabidopsis was used to transform flax (Linum usitatissimum) hypocotyl tissue. Transgenic regenerants were recovered from the inoculated tissue and were tested for expression of the foreign genes by leaf callus assays on kanamycin and on chlorsulfuron. Transgenic plants were grown to maturity; selfed progeny were similarly tested to determine segregation pattern for the novel genes, and some were grown in chlorsulfuron containing soil. Lines from two major commercial cultivars express chlorsulfuron resistance in greenhouse tests.

CDC Triffid transgenic flax

“…The gene was coupled to the bacterial-origin marker genes nopaline synthase and neomycin phosphotransferase II (NPT-II) in a plasmid and introduced into a disarmed Agrobacterium tumefaciens (McHughen 1989). Hypocotyl segments from NorLin flax were inoculated with the Agrobacterium in vitro, and shoots were regenerated, selected and maintained to maturity; details of the process are presented in McHughen 1989.…”

Tolerância de trigo (Triticum aestivum) e aveia (Avena sp.) a herbicidas inibidores da enzima acetolactato sintase (ALS)

“…Em algumas espé-cies de importância agrícola (milho, canola, beterraba, arroz, trigo e soja) foram desenvolvidas constituições genéticas para tolerância aos inibidores da ALS por meio de seleção de células somáticas, melhoramento por mutação, transformação genética e cruzamento interespecífico (Anderson & Georgeson, 1989;McHughen, 1989;Swanson et al, 1989;D'Halluin et al, 1992;Newhouse et al, 1992;Silva & Silva, 2007).…”

Novel Method to Detect a Construct-Specific Sequence of the Acetolactate Synthase Gene in Genetically-Modified Flax CDC Triffid (FP967)

“…Because some sulfonylurea herbicides are very persistent in soil, the choice of crops for use in rotations is limited. 1) To address this problem, McHughen developed a sulfonylurea-resistant flax line by introducing acetolactate synthase (ALS) gene cloned from Arabidopsis thaliana using Agrobacterium-based transformation. [1][2][3] The 5.8-kb ALS gene construct contains the ALS gene and its flanking regions.…”

“…1) To address this problem, McHughen developed a sulfonylurea-resistant flax line by introducing acetolactate synthase (ALS) gene cloned from Arabidopsis thaliana using Agrobacterium-based transformation. [1][2][3] The 5.8-kb ALS gene construct contains the ALS gene and its flanking regions. The coding sequence of the gene is modified by a single cytosine-to-thymine substitution at the 589th nucleotide, which results in reduced affinity for sulfonylurea herbicides.…”

“…The coding sequence of the gene is modified by a single cytosine-to-thymine substitution at the 589th nucleotide, which results in reduced affinity for sulfonylurea herbicides. 1) As well as the ALS gene, the construct also contains a neomycin phosphotransferase II gene (nptII), a spectinomycin/streptomycin resistance gene (Spec), a b-lactamase gene (bla), and the nopaline synthase gene (NOS). Expression of the resulting sulfonylurea herbicides resistance trait was subsequently demonstrated in progeny, and the transgenic line was designated CDC Triffid (FP967).…”