Abstract
Maize plants resistant to imidazolinone herbicides were engineered through targeted modification of endogenous genes using chimeric RNA/DNA oligonucleotides. A precise single-point mutation was introduced into genes encoding acetohydroxyacid synthase (AHAS), at a position known to confer imidazolinone resistance. Phenotypically normal plants from the converted events (C0) were regenerated from resistant calli and grown to maturity. Herbicide leaf painting confirmed the resistance phenotype in C0 plants and demonstrated the anticipated segregation pattern in C1 progeny. DNA cloning and sequencing of the targeted region in resistant calli and derived C0 and C1 plants confirmed the expected mutation. These results demonstrate that oligonucleotide-mediated gene manipulation can be applied to crop improvement. This approach does not involve genomic integration of transgenes. Since the new trait is obtained through modifying a gene within its normal chromosomal context, position effects, transgene silencing, or other concerns that arise as part of developing transgenic events are avoided.
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Acknowledgements
We thank Donglong Liu and Susan Jayne for advice on leaf painting and herbicide treatment, Marjorie Rudert, Toni Lockwood, Jacob Simmons, Michelle Siegrist, William Van Zante, the Pioneer Hi-Bred media prep and greenhouse staff for excellent technical assistance. We also thank Wesley Bruce, Susan Jayne, and Roger Kemble for reviewing this manuscript.
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Zhu, T., Mettenburg, K., Peterson, D. et al. Engineering herbicide-resistant maize using chimeric RNA/DNA oligonucleotides. Nat Biotechnol 18, 555–558 (2000). https://doi.org/10.1038/75435
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DOI: https://doi.org/10.1038/75435
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