Agrobacterium-mediated transformation of thin cell layer explants from Brassica napus L. |
| |
Authors: | P J Charest L A Holbrook J Gabard V N Iyer B L Miki |
| |
Institution: | (1) Department of Biology, Carleton University, K1S 5B6 Ottawa, Ontario, Canada;(2) Agriculture Canada, Plant Research Centre, K1A 0C6 Ottawa, Ontario, Canada;(3) Present address: Biotechnica International of Canada Inc., 170-6815 8 Street NE, T2E 7H7 Calgary, Alberta, Canada |
| |
Abstract: | Summary
Agrobacterium-mediated transformation of thin cell layer explants (Klimaszewska and Keller 1985) yielded large numbers of transgenic plants of a major Canadian rapeseed cultivar Brassica napus ssp. oleifera cv Westar. The morphology and fertility of these plants were indistinguishable from controls. The Ti plasmid vector, pGV3850 (Zambryski et al. 1983) was used as a cis vector and as a helper plasmid for the binary vector pBin19 (Bevan 1984). Selectable marker genes that conferred resistance to high levels of kanamycin (Km) on Nicotiana tabacum were less efficient in the selection of transgenic B. napus. At low levels of Km (15 g/ml) large numbers of transgenic plants (50%) were identified among the regenerants by nopaline synthase activity and several of these were confirmed by Southern blot analyses. Only a small number were resistant to higher levels of Km (80 g/ml). Preliminary analyses indicated that resistance to Km was transmitted to the selfed progeny. Chimeric chloramphenicol acetyl transferase genes were ineffective biochemical markers in transgenic B. napus.Contribution No. 1092 Plant Research Centre, Ontario, Canada |
| |
Keywords: | Brassica napus Rapeseed Transformation Agrobacterium tumefaciens Ti plasmid |
本文献已被 SpringerLink 等数据库收录! |
|