J Integr Plant Biol ›› 2023, Vol. 65 ›› Issue (11): 2416-2420.DOI: 10.1111/jipb.13567

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  • 收稿日期:2023-04-26 接受日期:2023-09-11 出版日期:2023-11-01 发布日期:2023-11-23

Efficient CRISPR/Cas9-mediated genome editing in sheepgrass (Leymus chinensis)

Zhelong Lin1†, Lei Chen1†, Shanjie Tang1, Mengjie Zhao1, Tong Li1, Jia You2, Changqing You1, Boshu Li3, Qinghua Zhao1, Dongmei Zhang2, Jianli Wang2, Zhongbao Shen2, Xianwei Song1, Shuaibin Zhang1* and Xiaofeng Cao1*   

  1. 1. State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, The Chinese Academy of Sciences, Beijing 100101, China;
    2. Institute of Forage and Grassland Sciences, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China;
    3. State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, The Chinese Academy of Sciences, Beijing 100101, China
    †These authors contributed equally to this work.
    *Correspondences:Shuaibin Zhang(sbzhang@genetics.ac.cn);Xiaofeng Cao(xfcao@genetics.ac.cn;Dr. Cao is fully responsible for the distributions of all materials associated with this article)
  • Received:2023-04-26 Accepted:2023-09-11 Online:2023-11-01 Published:2023-11-23

Abstract: The lack of genome editing platforms has hampered efforts to study and improve forage crops that can be grown on lands not suited to other crops. Here, we established efficient Agrobacterium-mediated clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) genome editing in a perennial, stress-tolerant forage grass, sheepgrass (Leymus chinensis). By screening for active single-guide RNAs (sgRNAs), accessions that regenerate well, suitable Agrobacterium strains, and optimal culture media, and co-expressing the morphogenic factor TaWOX5, we achieved 11% transformation and 5.83% editing efficiency in sheepgrass. Knocking out Teosinte Branched1 (TB1) significantly increased tiller number and biomass. This study opens avenues for studying gene function and breeding in sheepgrass.

Key words: Agrobacterium, genetic transformation, genome editing, LcTB1, sheepgrass

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