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Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing

Nature Plants volume 5, pages 480–485 (2019)Cite this article

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Abstract

Developing herbicide-tolerant varieties by genome editing holds great promise for addressing the worsening weed problems in wheat cultivation1. Here, we generated transgene-free wheat germplasms harbouring herbicide tolerance mutations that confer tolerance to sulfonylurea-, imidazolinone- and aryloxyphenoxy propionate-type herbicides by base editing the acetolactate synthase (ALS) and acetyl-coenzyme A carboxylase genes. These stackable herbicide tolerance traits provide a potentially powerful tool for weed management. In addition, we found that base editing at the wheat ALS Pro-174 codon (TaALS-P174) endowed wheat with sufficient resistance to nicosulfuron herbicide in MS growth medium to allow selection. When the TaALS-P174 editor was coupled with editors for other targets of interest, co-editing occurred in the nicosulfuron-resistant plants, and selection for resistance in growth medium enriched the frequency of coupled targets by several-fold. This selectable co-editing system has the potential to greatly bolster adoption of base editing for crop improvement applications.

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Fig 1: Multiallelic editing of TaALS-P174 codons endowed wheat tolerance to multiple herbicides.
Fig. 2: The TaALS-P174-based co-editing strategy efficiently generates mutations at desired positions via nicosulfuron selection.

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Data availability

All data supporting the findings of this study are available in the article or its Supplementary Information, or are available from the corresponding author upon reasonable request. Sequence data in this article can be found in the Ensembl Genomes database (http://plants.ensembl.org/Triticum_aestivum/Info/Index) under the following accession codes: TaALS-A (TraesCS6A02G288000), TaALS-B (TraesCS6B02G317400), TaALS-D (TraesCS6D02G268700), TaACCase-A (TraesCS2A02G069400), TaACCase-B (TraesCS2B02G082500) and TaACCase-D (TraesCS2D02G068100)

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Acknowledgements

This work was supported by grants from the National Key Research and Development Program of China (2016YFD0101804), the National Natural Science Foundation of China (31788103, 31471785, 31872933 and 31570369), the Chinese Academy of Sciences (QYZDY-SSW-SMC030 and KFJ-STS-ZDTP-024) and Beijing Municipal Science and Technology (D171100007717001 and Z171100001517001).

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Author notes

  1. These authors contributed equally: Rui Zhang, Jinxing Liu.

Authors and Affiliations

  1. State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China

    Rui Zhang, Jinxing Liu, Zhuangzhuang Chai, Sha Chen, Yang Bai, Yuan Zong, Kunling Chen & Caixia Gao

  2. University of Chinese Academy of Sciences, Beijing, China

    Zhuangzhuang Chai, Sha Chen, Yang Bai, Yuan Zong, Jiayang Li & Caixia Gao

  3. State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China

    Jiayang Li

  4. Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing, China

    Linjian Jiang

  5. MOA Key Lab of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, Beijing, China

    Linjian Jiang

Authors
  1. Rui Zhang
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Contributions

C.G., L.J. and J. Li designed the experiments; R.Z., Z.C., S.C. and Y.B. performed most of the experiments; J. Liu generated mutant plants. Y.Z. and K.C. analysed the results; C.G., L.J. and J. Li supervised the project; C.G., L.J., J. Li and R.Z. wrote the manuscript.

Corresponding authors

Correspondence to Jiayang Li, Linjian Jiang or Caixia Gao.

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Competing interests

C.G. and L.J. are inventors on patent applications covering generation of herbicide torlerance traits in wheat described in this work. C.G., R.Z. and J. Liu are inventors on patent applications describing generation of a selectable marker in wheat using base editing.

Additional information

Journal peer review information: Nature Plants thanks Donald Weeks, Bing Yang and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figures 1–8 and Supplementary Tables 1–13.

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Zhang, R., Liu, J., Chai, Z. et al. Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing. Nat. Plants 5, 480–485 (2019). https://doi.org/10.1038/s41477-019-0405-0

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