反应详情

To produce transgenic oilseed rape plants binary vectors in Agrobacterium tumefaciens C58C1:pGV2260 or Escherichia coli were used (Deblaere et al, 1984, Nucl. Acids. Res. 13, 4777-4788). For transforming oilseed rape plants (var. Drakkar, NPZ Nordeutsche Pflanzenzucht, Hohenlieth, Germany) a 1:50 dilution of an overnight culture of a positively transformed agrobacteria colony in Murashige-Skoog medium (Murashige and Skoog 19862 Physiol. Plant. 15, 473) containing 3% of saccharose (3MS medium) was used. Petioles or hypocotyledons of freshly germinated sterile rape plants (approx 1 cm2 each) were incubated in a Petri dish with a 1:50 agrobacteria dilution for 5-10 minutes. This was followed by 3-day concubation in darkness at 25° C. on 3MS medium containing 0.8% of Bacto-Agar. After three days, culturing was continued with 16 hours of light/8 hours of darkness and in a weekly cycle on MS medium containing 500 mg/l of Claforan (sodium cefotaxime), 50 mg/l of kanamycin, 20 microM of benzylaminopurine (BAP) and 1.6 g/l of glucose. Growing shoots were transferred onto MS medium containing 2% of saccharose, 250 mg/l of Claforan and 0.8% of Bacto-Agar. If after three weeks no roots had formed 2-indolylbutyric acid was added to the medium as a growth hormone for rooting purposes.

1. customTo produce transgenic oilseed rape plants binary vectors in Agrobacterium tumefaciens
2. customa 1:50 dilution of an overnight culture
3. waitThis was followed by 3-day
4. additionconcubation in darkness at 25° C. on 3MS medium containing 0.8% of Bacto-Agar
5. waitAfter three days
6. waitculturing was continued with 16 hours of light/8 hours of darkness and in a weekly cycle on MS medium