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<p><i>Setaria viridis</i> is a key model specie for functional genomic studies in C4 grasses and bioenergy crops. However, its genetic transformation remains challenging due to low efficiency and prolonged culture periods. This study presents an optimized transformation protocol employing a Thin Cell Layer (TCL) technique to induce morphogenic callus (MC) and enhance transformation rates. TCL-derived MC was co-cultured with <i>Agrobacterium tumefaciens</i> (AGL1 strain) carrying <i>uidA</i> (β-Glucuronidase) under the <i>Oryza sativa</i> Act1 promoter. Compared to the standard mature seed-derived callus method, TCL has the potential to reduce transformation time (19 to 15 weeks) and yield a transformation frequency of approximately 40%. The results present TCL as a robust alternative for efficient genetic transformation in <i>S. viridis</i>, facilitating its use in functional genomics.</p> Graphical Abstract <p></p>

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A new protocol for genetic transformation of Setaria viridis (L) using morphogenic callus via thin cell layers

  • 期刊论文

  • 发表日期：

    2025年9月10日

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  • Juan David Ferreira Gomes,
  • Juliana Nascimento Rodrigues,
  • Eveline Carla da Rocha Tavano,
  • Adriana Pinheiro Martinelli,
  • Marcio Alves-Ferreira

摘要

Setaria viridis is a key model specie for functional genomic studies in C4 grasses and bioenergy crops. However, its genetic transformation remains challenging due to low efficiency and prolonged culture periods. This study presents an optimized transformation protocol employing a Thin Cell Layer (TCL) technique to induce morphogenic callus (MC) and enhance transformation rates. TCL-derived MC was co-cultured with Agrobacterium tumefaciens (AGL1 strain) carrying uidA (β-Glucuronidase) under the Oryza sativa Act1 promoter. Compared to the standard mature seed-derived callus method, TCL has the potential to reduce transformation time (19 to 15 weeks) and yield a transformation frequency of approximately 40%. The results present TCL as a robust alternative for efficient genetic transformation in S. viridis, facilitating its use in functional genomics.

Graphical Abstract