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1

Regeneration of winter oilseed rape and trials of transformation mediated by Agrobacterium tumefaciens

100%
Starzycki M., Starzycka E., Matuszczak M.
Biotechnologia
|
1999
|
issue 1
164-171
EN
The effects of one strain Agrobacterium tumefaciens LBA 4404/pBI 121, conditions of co-cultivation and growth regulators on regeneration were studied and trials of transformation were performed on winter oilseed rape Brassica napus L. cv. Lirajet, Valesca and Bor. Strong shoots regeneration was observed after co-cultivation without acetosyringone. Among antibiotics that were used in the experiments, kanamycin A and geneticin hampered whereas carbenicillin and rifampicin stimulated shoots regeneration. The presence of nptII gene was confirmed by PCR and K1, K2 primers which generate a fragment of 732 bp in length.
2

Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by using Agrobacterium tumefaciens

75%
Cegielska-Taras T., Pniewski T., Szala L.
Journal of Applied Genetics
|
2008
|
vol. 49
|
issue 4
343-347
EN
Haploid microspore-derived embryos (MDEs) constitute a unique material for the introduction of new traits into winter oilseed rape (Brassica napus). MDEs have been transformed by using Agrobacterium tumefaciens strains EHA105 and LBA4404, both carrying the binary vector pKGIB containing the uidA gene encoding -glucuronidase (GUS) and the bar gene as a marker of resistance to phosphinotricin. Transformed embryos expressed GUS and regenerated plants that were resistant to herbicide Basta, as confirmed by a leaf-painting test. Progeny plants of the transformant T-39 were all transgenic, as they inherited T-DNA from their doubled haploid parental plant. Southern-blot analysis confirmed the integration and transmission of T-DNA into T1 plants. Transformation of MDEs facilitates the obtaining of winter oilseed rape homozygous for the introduced genes.
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