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The use of phosphomannose isomerase selection system for Agrobacterium-mediated transformation of tobacco and flax aimed for phytoremediation

J. Hilgert, M. Sura-De Jong, J. Fišer, K. Tupá, M. Vrbová, M. Griga, T. Macek, J. Žiarovská,

. 2017 ; 52 (5) : 338-345. [pub] 20170228

Language English Country England, Great Britain

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc17030979

A plant selection system based on the phosphomannose isomerase gene (pmi) as a selectable marker is often used to avoid selection using antibiotic resistance. Nevertheless, pmi gene is endogenous in several plant species and therefore difficult to use in such cases. Here we evaluated and compared Agrobacterium-mediated transformation of Linum usitatissimum breeding line AGT-952 (without endogenous pmi gene) and Nicotiana tabacum var. WSC-38 (with endogenous pmi gene). Transformation was evaluated for vectors bearing transgenes that have the potential to be involved in improved phytoremediation of contaminated environment. Tobacco regenerants selection resulted in 6.8% transformation efficiency when using a medium supplemented with 30 g/L mannose with stepwise decrease of the sucrose concentration. Similar transformation efficiency (5.3%) was achieved in transformation of flax. Relatively low selection efficiency was achieved (12.5% and 34.8%, respectively). The final detection of efficient pmi selection was conducted using PCR and the non-endogenous genes; pmi transgene for flax and todC2 transgene for tobacco plants.

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$a Hilgert, Jitka $u a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
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$a Sura-De Jong, Martina $u a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
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$a Fišer, Jiří $u a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
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$a Tupá, Kateřina $u a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
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$a Vrbová, Miroslava $u b Plant Biotechnology Department , AGRITEC Plant Research Ltd. , Šumperk , Czech Republic.
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$a Griga, Miroslav $u b Plant Biotechnology Department , AGRITEC Plant Research Ltd. , Šumperk , Czech Republic.
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$a Macek, Tomáš $u a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
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$a Žiarovská, Jana $u c Department of Genetics and Plant Breeding , University of Agriculture in Nitra , Slovak Republic.
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