Optimization of Brassica napus (canola) explant regeneration for genetic transformation

  1. Get@NRC: Optimization of Brassica napus (canola) explant regeneration for genetic transformation (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1016/j.nbt.2011.06.014
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Journal titleNew Biotechnology
Pages144155; # of pages: 12
SubjectAgrobacterium; Benzyladenine; Binary vectors; Brassica napus; Commercial productions; Dinitrophenylhydrazine; Donor plants; Firefly luciferase; Genetic transformation; Genetically modified; Hypocotyl explants; Luciferase assays; Microspore culture; Oil seed crops; Plantlets; Regeneration capacity; Regeneration medium; Reporter gene; Somatic embryo; Transgenesis; Cell culture; Crops; Cultivation; Genes; Nucleic acids; Plants (botany); 2,4 dinitrophenylhydrazine; 6 n benzyladenine; luciferase; article; bacterial strain; controlled study; genetic transformation; nonhuman; plant regeneration; priority journal; rapeseed; Rhizobium radiobacter; somatic embryogenesis; Agriculture; Agrobacterium tumefaciens; Brassica napus; Crops, Agricultural; Gene Transfer Techniques; Genetic Engineering; Hypocotyl; Plant Shoots; Regeneration; Transformation, Genetic; Agrobacterium; Brassica napus; Brassica napus var. napus
AbstractBrassica napus (canola) is the second largest oilseed crop in the world. It is among the first crops to be genetically transformed, and genetically modified cultivars are in commercial production at very significant levels. Despite the early lead with respect to transgenesis, there remain cultivars that are recalcitrant to transformation. To address this, we have conducted an elaborate investigation of the conditions for regenerating shoots from hypocotyl explants from four genetic lines: Invigor 5020, Westar and Topas as well as a microspore culture derived line of Topas (Line 4079). We analyzed the effect of hormonal combinations in regeneration medium, donor plant age and explant type on the regeneration capacity of these plants. The analysis showed that hypocotyls of eight-day-old seedlings grown on media supplemented with 1. mg/L dinitrophenylhydrazine (2,4-D) produced the most shoots. Globular somatic embryos emerged following two weeks of 2,4-D treatment. When transferred to the medium containing 5. mg/L benzyladenine (BA), approximately 82% of embryos produced shoots within six weeks. Invigor plants were shown to regenerate more efficiently than Topas; the number of plantlets regenerated from Invigor was approximately 40-50% more as compared to Topas or Line 4079. When hypocotyl explants were co-cultivated with the Agrobacterium strain GV3101 harboring a binary vector carrying a firefly luciferase reporter gene (LUC), significant numbers of plantlets were LUC-positive in a luciferase assay. Frequency of such plants were: Invigor 5020 (54.2 ± 2.5%), Westar (53.7 ± 5.3), Topas (16.0 ± 0.24) and Line 4079 (13.4 ± 4). © 2011 Elsevier B.V.
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AffiliationNational Research Council Canada (NRC-CNRC); NRC Plant Biotechnology Institute (PBI-IBP)
Peer reviewedYes
NPARC number21271390
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Record identifier598c1783-32df-48bc-9ad6-a32eeab11abb
Record created2014-03-24
Record modified2016-05-09
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