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

被引:40
作者
Kunze, I
Ebneth, M
Heim, U
Geiger, M
Sonnewald, U
Herbers, K
机构
[1] SunGene GmbH & Col KGaA, D-06466 Gatersleben, Germany
[2] Inst Pflanzengenet & Kulturpflanzenforsch, D-06466 Gatersleben, Germany
关键词
alternative selection marker; 2-deoxyglucose resistance; plant transformation;
D O I
10.1023/A:1011307508252
中图分类号
S3 [农学(农艺学)];
学科分类号
0901 ;
摘要
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 DOG(R) 1 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/12-DOG for tobacco. potato and tomato plants. After Agrobacterium tumefaciens-mediated transformation cells expressing the DOG(R) 1 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 DOG(R) 1 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 DOG(R) 1-expressing plants support the use of this selection system in crop plant transformation.
引用
收藏
页码:221 / 227
页数:7
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