Development of transformation vectors based upon a modified plant alpha-tubulin gene as the selectable marker.
A plant transformation and selection system has been developed utilizing a modified tubulin gene as a selectable marker. The vector constructs carrying a mutant alpha-tubulin gene from goosegrass conferring resistance to dinitroaniline herbicides were created for transformation of monocotyledonous and dicotyledonous plants. These constructs contained beta- and/or mutant alpha-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 microM for Eleusine coracana, Glycine max, Nicotiana plumbaginifolia and Nicotiana sylvestris; 3 microM for Linum usitatissimum. PCR and Southern blotting analyses of transformed lines with a specific probe to nptII, alpha-tubulin or beta-tubulin genes were performed to confirm the transgenic nature of regenerated plants. Band specific for the mutant alpha-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.
Yemets
Alla
A
Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kiev, Ukraine. alyemets@univ.kiev.ua
Radchuk
Vladimir
V
Bayer
Oleg
O
Bayer
Galina
G
Pakhomov
Alexey
A
Vance Baird
W
W
Blume
Yaroslav B
YB
D017343:Genes, Plant / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D053209:Herbicide Resistance / Q000235:genetics; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D012641:Selection, Genetic; D014170:Transformation, Genetic / Q000235:genetics*; D014274:Trifluralin / Q000494:pharmacology; D014404:Tubulin / Q000235:genetics*; D050257:Tubulin Modulators / Q000494:pharmacology
D005819:Genetic Markers; D014404:Tubulin; D050257:Tubulin Modulators; D014274:Trifluralin