J Integr Plant Biol ›› 2024, Vol. 66 ›› Issue (8): 1688-1702.DOI: 10.1111/jipb.13664

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  • 收稿日期:2024-01-18 接受日期:2024-03-29 出版日期:2024-08-01 发布日期:2024-08-19

Establishment of genome-editing system and assembly of a near-complete genome in broomcorn millet

Yang Liu1†, Zixiang Cheng2†, Weiyao Chen1,3, Chuanyin Wu2, Jinfeng Chen1* and Yi Sui2*   

  1. 1. State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, the Chinese Academy of Sciences, Beijing 100101, China
    2. State Key Laboratory of Crop Gene Resources and Breeding, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    3. University of Chinese Academy of Sciences, Beijing 100049, China
    These authors contributed equally to this work.
    *Correspondence: Jinfeng Chen (chenjinfeng@ioz.ac.cn); Yi Sui (suiyi@caas.cn, Dr. Sui is fully responsible for the distributions of all materials associated with this article)
  • Received:2024-01-18 Accepted:2024-03-29 Online:2024-08-01 Published:2024-08-19
  • Supported by:
    This work was funded by grants from the National Key Research and Development Program of China (2022YFD1400800; 2021YFF1000101‐4), the Innovation Program of the Chinese Academy of Agricultural Sciences, the National Natural Science Foundation of China (32172093), the China Postdoctoral Science Foundation funded project (2023M743843), and Initiative Scientific Research Program, Institute of Zoology, Chinese Academy of Sciences (2023IOZ0203).

Abstract: The ancient crop broomcorn millet (Panicum miliaceum L.) is an indispensable orphan crop in semi-arid regions due to its short life cycle and excellent abiotic stress tolerance. These advantages make it an important alternative crop to increase food security and achieve the goal of zero hunger, particularly in light of the uncertainty of global climate change. However, functional genomic and biotechnological research in broomcorn millet has been hampered due to a lack of genetic tools such as transformation and genome-editing techniques. Here, we successfully performed genome editing of broomcorn millet. We identified an elite variety, Hongmi, that produces embryogenic callus and has high shoot regeneration ability in in vitro culture. We established an Agrobacterium tumefaciens-mediated genetic transformation protocol and a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated genome-editing system for Hongmi. Using these techniques, we produced herbicide-resistant transgenic plants and edited phytoene desaturase (PmPDS), which is involved in chlorophyll biosynthesis. To facilitate the rapid adoption of Hongmi as a model line for broomcorn millet research, we assembled a near-complete genome sequence of Hongmi and comprehensively annotated its genome. Together, our results open the door to improving broomcorn millet using biotechnology.

Key words: broomcorn millet, CRISPR/Cas9 editing, genome assembly, herbicide resistance, transformation system

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