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A bacterial haloalkane dehalogenase (dhlA) gene as conditional negative selection marker for rice callus cells

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Abstract

The dhlA gene of Xanthobacter autotrophicus encodes dehalogenase that hydrolyzes dihaloalkanes such as 1,2-dichloroethane (DCE) into cytotoxic halogenated alcohol and an inorganic halide. As plants do not contain dehalogenase activity, they grow normally in the presence of DCE. We tested the transgenic expression of the bacterial dhlA gene in rice as a conditional negative selection marker. We developed 24 transgenic callus lines containing dhlA gene driven by rice actin-1 promoter, verified the expression of dhlA by Northern blot analysis, and subjected these transgenic lines to DCE treatment. We found that, while untransformed callus (Nipponbare) was unaffected by the DCE treatment, most of the transformed lines displayed symptoms of toxicity, indicating that dhlA is an effective conditional negative selection marker gene for rice in vitro cultures.

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Acknowledgments

This work was supported by funds received from USDA–CSREES and Division of Agriculture, University of Arkansas. The dhlA gene was obtained from David W. Ow.

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Authors and Affiliations

  1. Department of Crop, Soil and Environmental Sciences, University of Arkansas, Fayetteville, AR, 72701, USA

    Sarah K. Moore & Vibha Srivastava

  2. Department of Horticulture, University of Arkansas, Fayetteville, AR, 72701, USA

    Vibha Srivastava

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  1. Sarah K. Moore
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  2. Vibha Srivastava
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Correspondence to Vibha Srivastava.

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Editor: Gregory C. Phillips

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Moore, S.K., Srivastava, V. A bacterial haloalkane dehalogenase (dhlA) gene as conditional negative selection marker for rice callus cells. In Vitro Cell.Dev.Biol.-Plant 44, 468–473 (2008). https://doi.org/10.1007/s11627-008-9144-z

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  • DOI: https://doi.org/10.1007/s11627-008-9144-z

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