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Identification of herbicide resistance OsACC1 mutations via in planta prime-editing-library screening in rice

Nature Plants volume 7, pages 888–892 (2021)Cite this article

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Abstract

Base-editing-library-induced high density nucleotide substitutions have been applied to screen functional mutations in plants. However, due to limitations in the scope and conversion specificity of base editors, many desired mutations at pivotal protein sites may be overlooked. Here, we developed a prime-editing-library-mediated saturation mutagenesis (PLSM) method to substantially increase the diversity of amino acid substitutions at target sites for in planta screening. At six conserved residues of OsACC1, 16 types of herbicide-resistance-endowing mutations were identified. Most of these mutations exhibit reliable tolerance to aryloxyphenoxypropionate herbicides and have not been reported or applied in rice breeding. In addition, the advantage of PLSM was further shown by comparing the base-editing-mediated mutagenesis at the selected targets. The PLSM method established in this study has great potential for the direct evolution of genes related to agronomically important traits for crop improvement.

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Fig. 1: Screening herbicide-resistance mutations at pivotal sites of OsACC1 by PLSM.

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Data availability

All data in this study are available in the article or in the Supplementary Information, or are available from the corresponding author upon request. The NGS data can be accessed at the NCBI SRA database with accession code PRJNA705817. The locus number of OsACC1 is LOC_Os05g22940.

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Acknowledgements

This work was funded by the National Natural Science Foundation (grant nos. U19A2022 to P.W. and 32000284 to R.X.), Natural Science Foundation of Anhui province (grant nos. 2008085QC101 to R.X. and 2008085MC71 to R.Q.), Open Research Fund Program of Anhui Province Key Laboratory of Rice Genetics and Breeding (grant no. SDKF-2021-01 to J.L.) and Key Technology Research Project of Hefei (grant no. J2020G44 to P.W.).

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Author notes

  1. These authors contributed equally: Rongfang Xu, Xiaoshuang Liu.

Authors and Affiliations

  1. Key Laboratory of Rice Genetics & Breeding, Institute of Rice Research, Anhui Academy of Agricultural Science, Hefei, China

    Rongfang Xu, Xiaoshuang Liu, Juan Li, Ruiying Qin & Pengcheng Wei

  2. College of Agronomy, Anhui Agricultural University, Hefei, P. R. China

    Xiaoshuang Liu

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  1. Rongfang Xu
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  2. Xiaoshuang Liu
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  3. Juan Li
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Contributions

R.X. and J.L. contributed to the library constructions and genotyping. X.L. and R.Q. contributed to the rice transformations and herbicide treatments. P.W. conceived the study, supervised the research and wrote the manuscript with input from all authors.

Corresponding author

Correspondence to Pengcheng Wei.

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Competing interests

A patent application (no. 202110506568.0 to China) has been filed on the novel herbicide-resistant mutations.

Additional information

Peer review information Nature Plants thanks Sang-Gyu Kim, Bing Yang and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figs. 1–5 and Tables 1–3.

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Xu, R., Liu, X., Li, J. et al. Identification of herbicide resistance OsACC1 mutations via in planta prime-editing-library screening in rice. Nat. Plants 7, 888–892 (2021). https://doi.org/10.1038/s41477-021-00942-w

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