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Cyanamide hydratase as selectable marker in potato
Authors:Hella van Driel  Marina Keppel  Edwin van den Berg  Walter Schoo  Margaretha van der Lans  Marion Notkamp  Ellis Jansen  Eugene Roebroeck  Stephan Ohl  Sietske Stuiver-Hoekstra
Institution:1. Syngenta Mogen, Einsteinweg, Leiden, The Netherlands
2. Anthura BV, Anthuriumweg 14, 2665 kV, Bleiswijk, The Netherlands
3. Rijk Zwaan Breeding BV, Eerste kruisweg 9, 4793 RS, Fijnaart, The Netherlands
4. Monsanto Vegetable Seeds Division, P.O. Box 1050, 2660 BB, Bergschenhoek, The Netherlands
5. Lgem, Madepolderweg 45C, 2553 EG, The Hague, The Netherlands
6. Sigash, An der Althart 31, 67433, Neustadt Weinstrasse, Germany
Abstract:The selectable marker cyanamide hydratase/cyanamide was successfully used to generate transgenic potato plants from the cultivars Russet Burbank, Ranger Russet, Bintje, Desiree, Kardal and Pentland Dell. Up to 3,000 transgenics per person per year were produced. The efficiency of transgenic production varied among cultivars, and was in general 2–3 times lower than the transformation efficiency (TE) using the selectable marker kanamycin. Differences between cultivars in sensitivity to cyanamide selection were observed, but in general a concentration of 30 mg/l was applied for selection of transgenic shoots. A stepwise increase of cyanamide concentration during the transformation procedure for Russet Burbank resulted in an improved TE via a reduction of the escape efficiency from 69 to 29 %. The cultivars differed in the hormone concentration and duration (2,4-D and Zeatin riboside) required for the production of transgenics, predominantly during the phase of shoot initiation. Only for cultivars Bintje and Pentland Dell, adaptations in the hormone scheme are required during the transformation procedure with cyanamide selection compared to selection on kanamycin. Upon application of a transformation protocol for Russet Burbank, only in 1 % of the plants (in a population of 241) Agrobacterium could be detected, but these bacteria did not contain the vector for transgene transfer anymore.
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