Abstract
A reproducible system to produce transgenic Brassica napus plants has been developed using stem segments. Stem segments from 6–7 week old plants were inoculated with an Agrobacterium tumefaciens strain containing a disarmed tumor-inducing plasmid pTiT37-SE carrying a chimeric bacterial gene encoding kanamycin resistance (pMON200). Stem explants were cocultured for 2 days before transfer to kanamycin selection medium. Shoots regenerated directly from the explant in 3–6 weeks and were excised, dipped in Rootone®, and rooted in soil. Transformation was confirmed by opine production, kanamycin resistance, and DNA blot hybridization in the primary transformants. Final proof of transformation was demonstrated by the co-transfer of opine production and kanamycin resistance to progeny in a Mendelian fashion. Over 200 transgenic Brassica napus plants have been produced using this system.
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Abbreviations
- BA:
-
6-benzyladenine
- NAA:
-
α-naphthalene-acetic acid
- T-DNA:
-
transferred DNA into plants
- IBA:
-
indole butyric acid
- IAA:
-
indole acetic acid
- TXD:
-
Tobacco Xanthi diploid suspension cells
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Communicated by L. K. Grill
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Fry, J., Barnason, A. & Horsch, R.B. Transformation of Brassica napus with Agrobacterium tumefaciens based vectors. Plant Cell Reports 6, 321–325 (1987). https://doi.org/10.1007/BF00269550
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DOI: https://doi.org/10.1007/BF00269550