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Development of transformation vectors based upon a modified plant α‐tubulin gene as the selectable marker
Author(s) -
Yemets Alla,
Radchuk Vladimir,
Bayer Oleg,
Bayer Galina,
Pakhomov Alexey,
Baird W. Vance,
Blume Yaroslav B.
Publication year - 2008
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2007.11.012
Subject(s) - selectable marker , biology , transformation (genetics) , mutant , agrobacterium , gene , genetically modified crops , eleusine , transgene , tubulin , trifluralin , genetics , microbiology and biotechnology , microtubule , finger millet , weed control , agronomy
A plant transformation and selection system has been developed utilizing a modified tubulin gene as a selectable marker. The vector constructs carrying a mutant α‐tubulin gene from goosegrass conferring resistance to dinitroaniline herbicides were created for transformation of monocotyledonous and dicotyledonous plants. These constructs contained β‐ and/or mutant α‐tubulin genes driven either by ubiquitin or CaMV 35S promoter. The constructs were used for biolistic transformation of finger millet and soybean or for Agrobacterium ‐mediated transformation of flax and tobacco. Trifluralin, the main representative of dinitroaniline herbicides, was used as a selective agent in experiments to select transgenic cells, tissues and plantlets. Selective concentrations of trifluralin estimated for each species were as follows: 10 μM for Eleusine coracana, Glycine max, Nicotiana plumbaginifolia and Nicotiana sylvestris ; 3 μM for Linum usitatissimum . PCR and Southern blotting analyses of transformed lines with a specific probe to nptII , α‐tubulin or β‐tubulin genes were performed to confirm the transgenic nature of regenerated plants. Band specific for the mutant α‐tubulin gene was identified in transformed plant lines. Results confirmed the stable integration of the mutant tubulin gene into the plant genomes. The present study clearly demonstrates the use of a plant mutant tubulin as a selective gene for plant transformation.