Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew
Abstract
Sequence-specific nucleases have been applied to engineer targeted modifications in polyploid genomes, but simultaneous modification of multiple homoeoalleles has not been reported. Here we use transcription activator-like effector nuclease (TALEN) and clustered, regularly interspaced, short palindromic repeats (CRISPR)-Cas9 (refs. 4,5) technologies in hexaploid bread wheat to introduce targeted mutations in the three homoeoalleles that encode MILDEW-RESISTANCE LOCUS (MLO) proteins. Genetic redundancy has prevented evaluation of whether mutation of all three MLO alleles in bread wheat might confer resistance to powdery mildew, a trait not found in natural populations. We show that TALEN-induced mutation of all three TaMLO homoeologs in the same plant confers heritable broad-spectrum resistance to powdery mildew. We further use CRISPR-Cas9 technology to generate transgenic wheat plants that carry mutations in the TaMLO-A1 allele. We also demonstrate the feasibility of engineering targeted DNA insertion in bread wheat through nonhomologous end joining of the double-strand breaks caused by TALENs. Our findings provide a methodological framework to improve polyploid crops.
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CRISPR-Cas9 has been successfully applied to crop plants to alter agriculturally important traits such as disease resistance and drought tolerance.
“The technique has already been successfully applied to crop plants to alter agriculturally important traits such as disease resistance (Wang et al., 2014) and drought tolerance (Shi et al., 2017).”
From Genetically engineered crops help support conservation biological control (2019) · cites this paper as Wang et al., 2014
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- Authors
- Wang, Y., Cheng, X., Shan, Q., Zhang, Y., Liu, J., Gao, C., & Qiu, J.-L.
- Affiliations
- Chinese Academy of Sciences (CN)
- Corresponding author
- Gao, Caixia
- Qiu, Jin-Long
- Venue
- Nature Biotechnology
- Published
- 2014-07-20
- Volume
- 32
- Issue
- 9
- Pages
- 947-951
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W1980201684
- Cited by
- 2,072
- References
- 31
- Citations by year
- 2026: 114 · 2025: 174 · 2024: 163 · 2023: 212 · 2022: 245 · 2021: 235 · 2020: 203 · 2019: 170 · 2018: 145 · 2017: 173 · 2016: 136 · 2015: 82 · 2014: 18
- FWCI
- 56.00
- Type
- journal article
- Language
- en
- Update notices
- correction (2025-11-03)
- Open access
- closed
- Licence
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Chinese Academy of Sciences
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Wheat and Barley Genetics and Pathology
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Genetic and Mutation Studies
- Keywords
- hexaploid wheat, MLO proteins, powdery mildew resistance, TALEN, CRISPR/Cas9, genome editing, gene redundancy, polyploid crops, non-homologous end joining, transgenic wheat, broad-spectrum disease resistance
- MeSH terms
- Alleles, Ascomycota, Base Sequence, Molecular Sequence Data, Sequence Homology, Nucleic Acid, Triticum, RNA Editing
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