title: The use of phosphomannose isomerase selection system for Agrobacterium-mediated transformation of tobacco and flax aimed for phytoremediation.
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.
authors.affiliation: a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
authors.forename: Jitka
authors.initials: J
authors.lastname: Hilgert
authors.affiliation: a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Sura-De Jong
authors.affiliation: a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Fišer
authors.affiliation: a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Tupá
authors.affiliation: b Plant Biotechnology Department , AGRITEC Plant Research Ltd. , Šumperk , Czech Republic.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Vrbová
authors.affiliation: b Plant Biotechnology Department , AGRITEC Plant Research Ltd. , Šumperk , Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Griga
authors.affiliation: a Department of Biochemistry and Microbiology , University of Chemistry and Technology , Prague , Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Macek
authors.affiliation: c Department of Genetics and Plant Breeding , University of Agriculture in Nitra , Slovak Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Žiarovská
mesh_terms: D060054:Agrobacterium / Q000235:genetics*; D001673:Biodegradation, Environmental; D003470:Culture Media / Q000737:chemistry; D019597:Flax / Q000187:drug effects / Q000235:genetics*; D008358:Mannose / Q000378:metabolism / Q000494:pharmacology; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D012641:Selection, Genetic; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics*; D014169:Transformation, Bacterial / Q000235:genetics*
chemical_list: D003470:Culture Media; D008359:Mannose-6-Phosphate Isomerase; D008358:Mannose
keywords: Linum usitatissimum; nicotiana tabacum; phosphomannose isomerase; positive selection system