biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 52:453-461, 2008 | DOI: 10.1007/s10535-008-0090-8

Use of phosphomannose isomerase-based selection system for Agrobacterium-mediated transformation of tomato and potato

J. Bříza1,*, D. Pavingerová1, P. Přikrylová2, J. Gazdová2, J. Vlasák1, H. Niedermeierová1
1 Institute of Plant Molecular Biology, Biology Centre of the Academy of Sciences of the Czech Republic, České Budějovice, Czech Republic
2 Faculty of Science, University of South Bohemia, České Budějovice, Czech Republic

Two selection systems for Agrobacterium tumefaciens mediated transformation of tomato and potato were compared. In the tomato (Lycopersicon esculentum 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 dm-3. The best transformation efficacy with the commonly used concentration of 100 mg dm-3 kanamycin as a selection agent was 9 %. In the potato (Solanum tuberosum cv. Bintje), the highest transformation frequency was 53.3 % when mannose concentration in the regeneration medium was 5 g dm-3 during the first 3 weeks after transformation and 10 g dm-3 afterwards. The optimum concentration of sucrose was 20 g dm-3. The transformation efficiency using kanamycin as a selection agent at a concentration 100 mg dm-3 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.

Keywords: Lycopersicon esculentum; npt II; pmi; Solanum tuberosum; transformation efficiency
Subjects: Agrobacterium tumefaciens; chromosomes; genetic transformation; in vitro culture, growth control; in vitro culture, regeneration; in vitro culture, rooting; Lycopersicon esculentum; nutrient medium, Murashige and Skoog (MS); plasmid; potato; Solanum tuberosum; tomato

Received: July 2, 2007; Accepted: December 17, 2007; Published: September 1, 2008  Show citation

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Bříza, J., Pavingerová, D., Přikrylová, P., Gazdová, J., Vlasák, J., & Niedermeierová, H. (2008). Use of phosphomannose isomerase-based selection system for Agrobacterium-mediated transformation of tomato and potato. Biologia plantarum, 52(3), 453-461. doi: 10.1007/s10535-008-0090-8
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