Journal
PLANT CELL TISSUE AND ORGAN CULTURE
Volume 99, Issue 1, Pages 97-108Publisher
SPRINGER
DOI: 10.1007/s11240-009-9580-4
Keywords
Sorghum bicolor; Agrobacterium; bar; Binary vector; Marker-free; Transformation
Funding
- University of Missouri Agricultural Experiment Station
- Provost's office
- University of Missouri Food for the twenty-first Century Eminence Program.
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We report an Agrobacterium-mediated transformation system that can generate marker-free transgenic sorghum [Sorghum bicolor (L.) Moench] from a public line [P898012] using standard binary vectors with bar as a selectable marker. Eight co-cultivation conditions were examined for their effect on transformation. The average transformation frequencies were 0.4 and 0.7% for pZY101-TC2 and pZY101-SKRS, respectively, derived from binary vector pZY102 and containing bar and target gene(s) in separate T-DNA regions. A low selection pressure (2.5 mg l(-1) DL-phosphinothrithin, PPT) was deployed during callus induction in combination with rapid selection to generate plants from 80 independent events, all but three of which were fertile and set seed. PCR and Southern analyses showed that 36 out of 80 events contained both bar and the target gene(s) (an average co-transformation frequency of 45%). Seedlings of the T1 generation transmitted T-DNAs with target gene(s) and bar gene independently, generating a fraction of progeny with only the target gene(s).
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