The use of phosphomannose isomerase selection system for Agrobacterium-mediated transformation of tobacco and flax aimed for phytoremediation |
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Authors: | Jitka Hilgert Martina Sura-De Jong Ji?í Fi?er Kate?ina Tupá Miroslava Vrbová Miroslav Griga |
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Institution: | 1. Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic;2. Plant Biotechnology Department, AGRITEC Plant Research Ltd., ?umperk, Czech Republic |
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Abstract: | 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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Keywords: | Linum usitatissimum nicotiana tabacum phosphomannose isomerase positive selection system |
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