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Overexpression of an Arabidopsis thaliana ABC transporter confers kanamycin resistance to transgenic plants

Nature Biotechnology volume 23, pages 1177–1180 (2005)Cite this article

A Corrigendum to this article was published on 01 October 2005

Abstract

Selectable markers of bacterial origin such as the neomycin phosphotransferase type II gene, which can confer kanamycin resistance to transgenic plants, represent an invaluable tool for plant engineering. However, since all currently used antibiotic-resistance genes are of bacterial origin, there have been concerns about horizontal gene transfer from transgenic plants back to bacteria, which may result in antibiotic resistance. Here we characterize a plant gene, Atwbc19, the gene that encodes an Arabidopsis thaliana ATP binding cassette (ABC) transporter and confers antibiotic resistance to transgenic plants. The mechanism of resistance is novel, and the levels of resistance achieved are comparable to those attained through expression of bacterial antibiotic-resistance genes in transgenic tobacco using the CaMV 35S promoter. Because ABC transporters are endogenous to plants, the use of Atwbc19 as a selectable marker in transgenic plants may provide a practical alternative to current bacterial marker genes in terms of the risk for horizontal transfer of resistance genes.

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Figure 1: Use of Atwbc19 as a selection marker.
Figure 2: Kanamycin resistance of T1 transgenic plants transformed with pABC.
Figure 3

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Acknowledgements

We are grateful to John Dunlap for his assistance in confocal microscopy imaging and Andreas Nebenführ for helpful discussions as well as improvement suggested by three anonymous reviewers. We wish to thank Megan York, Jason Burris, Jaime Davis and Reginald Millwood for assistance in lab and greenhouse work. This research work was supported by The University of Tennessee Institute of Agriculture and the US Army.

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Authors and Affiliations

  1. Department of Plant Sciences, The University of Tennessee, 252 Ellington Plant Sciences, 2431 Joe Johnson Drive, Knoxville, 37996-4561, Tennessee, USA

    Ayalew Mentewab & C Neal Stewart Jr

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  1. Ayalew Mentewab
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  2. C Neal Stewart Jr
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Correspondence to C Neal Stewart Jr.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Root growth and NPTII synthesis levels in control Arabidopsis thaliana Col1 and homozygous insertional knockout mutants (KO). (PDF 854 kb)

Supplementary Fig. 2

Twelve-week old non-transgenic and T1 ABC-transgenic tobacco plants in the greenhouse. (PDF 124 kb)

Supplementary Fig. 3

Root length distribution frequency of non-transgenic tobacco (Xanthi), and T1 transgenic lines 28 and 30 grown on MSO media with or without 100 or 200 mg/l kanamycin. (PDF 160 kb)

Supplementary Fig. 4

T-DNA region of construct used for subcellular localization of AtWBC19. (PDF 603 kb)

Supplementary Fig. 5

Clustal alignment of the Arabidopsis thaliana WBC members most closely related to Atwbc19. (PDF 394 kb)

Supplementary Table 1

Phenotypic data of 9-week-old greenhouse-grown T1 tobacco plants. (PDF 37 kb)

Supplementary Table 2

Transformation of canola (Brassica napus) cv. Westar hypocotyl segments with plasmids pNPT and pABC. (PDF 40 kb)

Supplementary Notes (PDF 92 kb)

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Mentewab, A., Stewart, C. Overexpression of an Arabidopsis thaliana ABC transporter confers kanamycin resistance to transgenic plants. Nat Biotechnol 23, 1177–1180 (2005). https://doi.org/10.1038/nbt1134

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