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Lysine racemase: a novel non-antibiotic selectable marker for plant transformation

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Abstract

A non-antibiotic based selection system using l-lysine as selection agent and the lysine racemase (lyr) as selectable marker gene for plant transformation was established in this study. l-lysine was toxic to plants, and converted by Lyr into d-lysine which would subsequently be used by the transgenic plants as nitrogen source. Transgenic tobacco and Arabidopsis plants were successfully recovered on l-lysine medium at efficiencies of 23 and 2.4%, respectively. Phenotypic characterization of transgenic plants clearly revealed the expression of normal growth and developmental characteristics as that of wild-type plants, suggesting no pleiotropic effects associated with the lyr gene. The specific activity of Lyr in transgenic tobacco plants selected on l-lysine ranged from 0.77 to 1.06 mU/mg protein, whereas no activity was virtually detectable in the wild-type plants. In addition, the composition of the free amino acids, except aspartic acid, was not affected by the expression of the lyr gene in the transgenic tobacco plants suggesting very limited interference with endogenous amino acid metabolism. Interestingly, our findings also suggested that the plant aspartate kinases may possess an ability to distinguish the enantiomers of lysine for feedback regulation. To our knowledge, this is the first report to demonstrate that the lysine racemase selectable marker system is novel, less controversial and inexpensive than the traditional selection systems.

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Acknowledgments

We thank Dr. Chang-Hsien Yang (Professor, Institute of Biotechnology, NCHU) for his generous gift of Arabidopsis thaliana (ecotype Columbia) seeds and Agrobacterium strain GV3101 and also providing the growth chamber facilities for the maintenance of Arabidopsis transformants. This work was supported by the grants from the Council of Agriculture (96 AS-1.1.1-FD-Z1) and National Science Council (NSC 92-2317-B-005-018), Taiwan.

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  1. Institute of Molecular Biology, College of Life Sciences, National Chung Hsing University, Taichung, 402, Taiwan

    I-Chieh Chen, Venkatesan Thiruvengadam, Wei-De Lin & Wen-Hwei Hsu

  2. Department of Medical Research, China Medical University Hospital, Taichung, 404, Taiwan

    Wei-De Lin

  3. Department of Plant Pathology, College of Agriculture and Natural Resources, National Chung Hsing University, Taichung, 402, Taiwan

    Ho-Hsiung Chang

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  1. I-Chieh Chen
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  2. Venkatesan Thiruvengadam
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  3. Wei-De Lin
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Correspondence to Wen-Hwei Hsu.

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I-Chieh Chen and Venkatesan Thiruvengadam contributed equally to this work.

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Chen, IC., Thiruvengadam, V., Lin, WD. et al. Lysine racemase: a novel non-antibiotic selectable marker for plant transformation. Plant Mol Biol 72, 153–169 (2010). https://doi.org/10.1007/s11103-009-9558-y

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