Use of phosphomannose isomerase-based selection system for Agrobacterium-mediated trasformation of tomato and potato
2008
Briza, J.,Branisovska 31, Ceske Budejovice (Czech Republic) | Pavingerova, D.,Akademie Ved, Ceske Budejovice (Czech Republic). Ustav Molekularni Biologie Rostlin | Prikrylova, P.,Jihoceska Univ., Ceske Budejovice (Czech Republic). Prirodovedecka Fakulta | Gazdova, J.,Jihoceska Univ., Ceske Budejovice (Czech Republic). Prirodovedecka Fakulta | Vlasak, J.,Akademie Ved, Ceske Budejovice (Czech Republic). Ustav Molekularni Biologie Rostlin | Niedermeierova, H.,Akademie Ved, Ceske Budejovice (Czech Republic). Ustav Molekularni Biologie Rostlin
Two selection systems for Agrobacterium tumefaciens mediated transformation of tomato and potato were compared. In the tomato (cv. Moneymaker), the highest transformation rate (4.2%) of cotyledon explants on mannose-selection medium was obtained when mannose/sucrose concentration in the regeneration medium was 5/15 g/cubic dm. The best transformation efficacy with the commonly used concentration of 100 mg/cubic dm kanamycin as a selection agent was 9%. In the potato (cv. Bintje), the highest transformation frequency was 53.3% when mannose concentration in the regeneration medium was 5 g/cubic dm during the first 3 weeks after transformation and 10 g/cubic dm afterwards. The optimum concentration of sucrose was 20 g/cubic dm. The transformation efficiency using kanamycin as a selection agent at a concentration 100 mg/cubic dm was 33.3% with potato. Our results demonstrate that the transformation efficiency using mannose selection is 1.6-fold higher for potato and about 2 times lower for tomato comparing with the ordinary protocol using kanamycin.
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