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Long-term assessment of transgene behavior in barley: Ds-mediated delivery of bar results in robust, stable, and heritable expression

  • Biotechnology/Genetic Transformation/Functional Genomics
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Abstract

The utility of transgenic plants for both experimental and practical agronomic purposes is highly dependent on stable, predictable, and heritable expression of the introduced genes. This requirement is frequently unfulfilled, and transgenes often are completely silenced. Studies of transgenic loci have shown that rearrangements of transgenes occur during the integration process, some of which are potent cues that induce silencing. Conversely, intact, single-copy transgenes produced via transposon-mediated gene delivery have shown relatively stable expression, at least in early-generation progeny. To examine the long-term expression stability of a bar expression cassette delivered via Dissociation (Ds)-mediated transposition, we examined qualitative and quantitative expression in barley (Hordeum vulgare L.) populations developed for transposon tagging. Qualitative assessments of herbicide resistance among 106 lines showed bar expression to be stable for at least five generations of advance via self-pollination. Similarly, qualitatively stable expression was observed among 31 near-isogenic lines derived from at least seven backcrosses to the cultivar Garnet. Quantitative RT-PCR measurements of bar expression were conducted for eight near-isogenic lines and their donor parents. The expression of bar was highly correlated in parent and progeny near-isogenic lines, showing high heritability of bar expression. These data demonstrate stable, predictable transgene expression following Ds-mediated delivery.

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Acknowledgments

This work was funded by the USDA-ARS CRIS project 5366-21000-028. The USDA-ARS is an equal opportunity employer.

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  1. Ryan H. Brown

    Present address: General Mills, 150 N. Research Dr., Kannapolis, NC, 28081, USA

Authors and Affiliations

  1. National Small Grains Germplasm Research Facility, USDA-ARS, 1691 S. 2700 W, Aberdeen, ID, 83210, USA

    Phil Bregitzer & Ryan H. Brown

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  1. Phil Bregitzer
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Correspondence to Phil Bregitzer.

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Editor: Todd Jones

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Bregitzer, P., Brown, R.H. Long-term assessment of transgene behavior in barley: Ds-mediated delivery of bar results in robust, stable, and heritable expression. In Vitro Cell.Dev.Biol.-Plant 49, 231–239 (2013). https://doi.org/10.1007/s11627-013-9507-y

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  • DOI: https://doi.org/10.1007/s11627-013-9507-y

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