2-Deoxyglucose resistance: a novel selection marker for plant transformation.

Autor: Kunze, Irene, Ebneth, Marcus, Heim, Ute, Geiger, Michael, Sonnewald, Uwe, Herbers, Karin
Zdroj: Molecular Breeding; Jun2001, Vol. 7 Issue 3, p221-227, 7p
Abstrakt: A novel selection marker for plant transformation alternative to antibiotic and herbicide resistance is described. The selective agent applied is 2-deoxyglucose (2-DOG) which in the cytosol of plant cells is phosphorylated by hexokinase yielding 2-DOG-6-phosphate (2-DOG-6-P). 2-DOG-6-P exerts toxic effects on overall cellular metabolism leading to cell death. We observed that constitutive expression of the yeast DOGR1 gene encoding a 2-DOG-6-P phosphatase resulted in resistance towards 2-DOG in transgenic tobacco plants. This finding was exploited to develop a selection system during transformation of tobacco and potato plants. The lowest concentration of 2-DOG leading to nearly complete inhibition of regeneration of wild-type explants was found to range between 400 and 600 mg/l 2-DOG for tobacco, potato and tomato plants. After Agrobacterium tumefaciens-mediated transformation cells expressing the DOGR1 gene were selected by resistance to 2-DOG. More than 50% of tobacco explants formed shoots and on average 50% of these shoots harboured the DOGR1 gene. Similar results were obtained for potato cv. Solara. The acceptability of the resistance gene derived from baker's yeast, the unobjectionable toxicological data of 2-DOG as well as the normal phenotype of DOGR1-expressing plants support the use of this selection system in crop plant transformation. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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