A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis.
The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1, 2-dichloroethane (DCE), to a halogenated alcohol and an inorganic halide (Janssen et al. 1994, Annu. Rev. Microbiol. 48, 163-191). In Xanthobacter, these alcohols are further catabolized by alcohol and aldehyde dehydrogenase activities, and by the product of the dhlB gene to a second halide and a hydroxyacid. The intermediate halogenated alcohols and, in particular, the aldehydes are more toxic than the haloalkane substrates or the pathway products. We show here that plants, including Arabidopsis, tobacco, oil seed rape and rice, do not express detectable haloalkane dehalogenase activities, and that wild-type Arabidopsis grows in the presence of DCE. In contrast, DCE applied as a volatile can be used to select on plates or in soil transgenic Arabidopsis which express dhlA. The dhlA marker therefore provides haloalkane dehalogenase reporter activity and substrate dependent negative selection in transgenic plants.
Department of Plant Phsiology, Molecular Biology Institue, Copenhagen University, Denmark.
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Naested
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Fennema
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Hao
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Andersen
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Janssen
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Mundy
D017360:Arabidopsis / Q000235:genetics*; D015946:Ethylene Dibromide / Q000378:metabolism; D005025:Ethylene Dichlorides / Q000378:metabolism; D005798:Genes, Bacterial; D005819:Genetic Markers; D016946:Gram-Negative Aerobic Rods and Cocci / Q000235:genetics; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics*; D030821:Plants, Genetically Modified; D012641:Selection, Genetic
D005025:Ethylene Dichlorides; D005819:Genetic Markers; D015946:Ethylene Dibromide; C024565:ethylene dichloride; D006867:Hydrolases; C046137:haloalkane dehalogenase