Poster: Emerging technologies
Abs #
911: Mutated plant tubulin genes as a novel system for transgenic plant selection: mode of action and ecological safety
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Presenter: |
Blume, Yaroslav B., cellbio@cellbio.freenet.viaduk.net |
Authors | Yemets, Alla I. (A) Radchuk, Vladimir V. (A) Baird, William V. (B) Blume, Yaroslav B. (A) | | Affiliations: |
(A): Dept. of Genomics and Biotechnology, Inst. of Cell Biol. and Genet. Engineering, Natl. Acad. of Sci. of Ukraine
(B): Dept. of Horticulture and Genetics, Clemson Univ.
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A selection system for plant transformation exclusively based on genetic information of plant origination is proposed. This alternative to antibiotic resistance genes is based on the developing knowledge about the mechanisms of plant microtubules resistance to antimitotic herbicides (dinitroanilines, DNH, and phosphorothioamidates, PTH), involving the appearance of altered tubulins as results of mutation(s) in the binding site for DNH/PTH. At current stage this knowledge includes:
i) Characterization of mutant α-tubulins from natural biotypes of goosegrass (Eleusine indica), which confer resistance to DNH;
ii) Production of Nicotiana plumbaginifolia mutants resistant to PTH and DNH (in both cases with altered β-tubulin)and cloning and sequencing of mutant β-tubulin;
iii) 3-D reconstruction of interactive sites of α- and β-tubulin for DNH and PTH of tertiary structure of sensitive and resistant tubulin molecules and predication of new possible sites for such interaction.
The panel of asymmetric somatic hybrids between β-tubulin mutants and other dicots species was produced to demonstrate a potential usage of altered tubulin as selective marker and its role in phenotypic expression of the resistance to DNH and PTH. Now we are working on:
i) Production of a selection marker cassettes with sequences coding for α-/β-tubulin conferring resistance to DNH and PTH and full-length β-/α-tubulin);
ii) Transformation of model plants with these constructs and establishment of the transformation and selection based on DNH/PTH.
