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Generation of a multi-herbicide-tolerant alfalfa by using base editing



We present the first report on base editing in alfalfa. Specifically, we showed edited alfalfa with tolerance to both sulfonylurea- and imidazolinone-type herbicides.

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  • Bottero E, Massa G, González M, Stritzler M, Tajima H, Gómez C, Frare R, Feingold S, Blumwald E, Ayub N, Soto G (2021) Efficient CRISPR/Cas9 genome editing in alfalfa using a public germplasm. Front Agron 3:661526

    Article  Google Scholar 

  • Chen H, Zeng Y, Yang Y, Huang L, Tang B, Zhang H, Hao F, Liu W, Li Y, Liu Y, Zhang X, Zhang R, Zhang Y, Li Y, Wang K, He H, Wang Z, Fan G, Yang H, Bao A, Shang Z, Chen J, Wang W, Qiu Q (2020) Allele-aware chromosome-level genome assembly and efficient transgene-free genome editing for the autotetraploid cultivated alfalfa. Nat Commun 11:2494

    CAS  Article  Google Scholar 

  • Dieterich Mabin ME, Brunet J, Riday H, Lehmann L (2021) Self-fertilization, inbreeding, and yield in alfalfa seed production. Front Plant Sci 12:700708

    Article  Google Scholar 

  • García AN, Ayub ND, Fox AR, Gómez MC, Diéguez MJ, Pagano EM, Berini CA, Muschietti JP, Soto G (2014) Alfalfa snakin-1 prevents fungal colonization and probably coevolved with rhizobia. BMC Plant Biol 14:248

    Article  Google Scholar 

  • Heap I, Duke SO (2018) Overview of glyphosate-resistant weeds worldwide. Pest Manag Sci 74:1040–1049

    CAS  Article  Google Scholar 

  • Jozefkowicz C, Frare R, Fox R, Odorizzi A, Arolfo V, Pagano E, Basigalup D, Ayub N, Soto G (2018) Maximizing the expression of transgenic traits into elite alfalfa germplasm using a supertransgene configuration in heterozygous conditions. Theor Appl Genet 131:1111–1123

    CAS  Article  Google Scholar 

  • Mishra R, Joshi RK, Zhao K (2020) Base editing in crops: current advances, limitations and future implications. Plant Biotechnol J 18:20–31

    Article  Google Scholar 

  • Samac DA, Temple SJ (2021) Biotechnology advances in Alfalfa. Compendium of plant genomes. Springer, Cham

    Google Scholar 

  • Veillet F, Perrot L, Chauvin L, Kermarrec M-P, Guyon-Debast A, Chauvin J-E, Nogué F, Mazier M (2019) Transgene-free genome editing in tomato and potato plants using agrobacterium-mediated delivery of a CRISPR/Cas9 cytidine base editor. Int J Mol Sci 20:402

    Article  Google Scholar 

  • Wolabu TW, Cong L, Park J-J, Bao Q, Chen M, Sun J, Xu B, Ge Y, Chai M, Liu Z, Wang Z-Y (2020) Development of a highly efficient multiplex genome editing system in outcrossing Tetraploid Alfalfa (Medicago sativa). Front Plant Sci 11:1063

    Article  Google Scholar 

  • Zhang R, Liu J, Chai Z, Chen S, Bai Y, Zong Y, Chen K, Li J, Jiang L, Gao C (2019) Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing. Nature Plants 5:480–485

    CAS  Article  Google Scholar 

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This work was supported by grants PICT 2017#0484, FVT 2017#39 and PICT-STARTUP 2020 #00004 provided to GS and by PE I115 (Biotechnology Program of INTA, Argentina). EB has a graduate student fellowship from CONICET, Argentina. MS was supported by fellowships from CONICET and IUBMB.

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Conceived and designed the experiments: MS and GS. Performed the experiments: EB, CG, MS, HT, RF, CP, NA and GS. Analyzed the data: EB, CG, MS, HT, EB, NA and GS. Wrote the paper: GS. All authors read and approved the final manuscript.

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Correspondence to Gabriela Soto.

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The authors declare that they have no conflict of interest.

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Communicated by Neal Stewart.

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Bottero, E., Gómez, C., Stritzler, M. et al. Generation of a multi-herbicide-tolerant alfalfa by using base editing. Plant Cell Rep 41, 493–495 (2022).

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  • Alfalfa
  • Base editing
  • CRISPR/Cas9
  • ALS
  • Herbicides