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

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Biologia Plantarum

Abstract

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.

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Abbreviations

MS:

Murashige and Skoog (1962) medium

NPT II:

neomycin phosphotransferase II

PMI:

phosphomannose isomerase

RM:

regeneration medium

SE:

selection efficiency

TE:

transformation efficiency

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Authors and Affiliations

  1. Institute of Plant Molecular Biology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, CZ-37005, České Budějovice, Czech Republic

    J. Bříza, D. Pavingerová, J. Vlasák & H. Niedermeierová

  2. Faculty of Science, University of South Bohemia, Branišovská 31, CZ-37005, České Budějovice, Czech Republic

    P. Přikrylová & J. Gazdová

Authors
  1. J. Bříza
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  2. D. Pavingerová
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  3. P. Přikrylová
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  4. J. Gazdová
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  5. J. Vlasák
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  6. H. Niedermeierová
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Correspondence to J. Bříza.

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Bříza, J., Pavingerová, D., Přikrylová, P. et al. Use of phosphomannose isomerase-based selection system for Agrobacterium-mediated transformation of tomato and potato. Biol Plant 52, 453–461 (2008). https://doi.org/10.1007/s10535-008-0090-8

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  • DOI: https://doi.org/10.1007/s10535-008-0090-8

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