A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis

Henrik Næsted; M. Fennema; L. Hao; M. Andersen; D.B. Janssen; John Williams Mundy

doi:10.1046/j.1365-313X.1999.00477.x

A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis

Henrik Næsted, M. Fennema, L. Hao, M. Andersen, D.B. Janssen, John Williams Mundy

Centre m.v.

26 Citations (Scopus)

Abstract

The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1,2-dichloroethane (DCE), to a halo- genated 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.

Original language	English
Journal	Plant Journal
Volume	18
Issue number	5
Pages (from-to)	571-576
ISSN	0960-7412
DOIs	https://doi.org/10.1046/j.1365-313X.1999.00477.x
Publication status	Published - 1999

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10.1046/j.1365-313X.1999.00477.x

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@article{562845d074c911dbbee902004c4f4f50,

title = "A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis ",

abstract = "The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1,2-dichloroethane (DCE), to a halo- genated 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.",

author = "Henrik N{\ae}sted and M. Fennema and L. Hao and M. Andersen and D.B. Janssen and Mundy, {John Williams}",

year = "1999",

doi = "10.1046/j.1365-313X.1999.00477.x",

language = "English",

volume = "18",

pages = "571--576",

journal = "Plant Journal",

issn = "0960-7412",

publisher = "Wiley-Blackwell",

number = "5",

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TY - JOUR

T1 - A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis

AU - Næsted, Henrik

AU - Fennema, M.

AU - Hao, L.

AU - Andersen, M.

AU - Janssen, D.B.

AU - Mundy, John Williams

PY - 1999

Y1 - 1999

N2 - The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1,2-dichloroethane (DCE), to a halo- genated 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.

AB - The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1,2-dichloroethane (DCE), to a halo- genated 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.

U2 - 10.1046/j.1365-313X.1999.00477.x

DO - 10.1046/j.1365-313X.1999.00477.x

M3 - Journal article

SN - 0960-7412

VL - 18

SP - 571

EP - 576

JO - Plant Journal

JF - Plant Journal

IS - 5

ER -

A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis

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