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Main conclusion <p>This study overcomes a key technical barrier by establishing transformation and enabling first CRISPR/Cas9 editing in <i>Oxalis</i>, providing a platform for functional genomics and breeding.</p> Abstract <p>The lack of an efficient genetic transformation system has considerably hindered functional genomics studies in <i>Oxalis vulcanicola</i> ‘Sunset Velvet’. Here, we established a stable and efficient <i>Agrobacterium</i>-mediated transformation system using stem segments as explants. Key parameters, including pre-culture duration, infection time, <i>Agrobacterium</i> cell density, acetosyringone (AS) concentration, and co-cultivation period, were systematically optimized. Under optimal conditions, the highest transient β-glucuronidase (GUS) expression rate reached approximately 9.0%, and eight stable transgenic lines were successfully obtained. CRISPR/Cas9-mediated genome editing was achieved in <i>Oxalis</i> for the first time. Targeted mutagenesis of <i>OvPDS1</i>, a gene involved in carotenoid biosynthesis, resulted in an albino phenotype, and Sanger sequencing confirmed a base substitution at the target site. Although the editing efficiency was relatively low (0.5%), this result demonstrates the feasibility of genome editing in <i>Oxalis</i>. This study overcomes a major technical bottleneck and provides a robust platform for functional gene analysis, trait improvement, and molecular breeding in <i>O. vulcanicola</i> ‘Sunset Velvet’ and other non-model ornamental plants.</p>

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Establishment of an efficient Agrobacterium-mediated transformation system and CRISPR/Cas9-mediated genome editing of the OvPDS1 gene in Oxalis vulcanicola ‘Sunset Velvet’

  • 期刊论文

  • 发表日期：

    2026年7月28日

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  • Wanli Tuo,
  • Xuexuan Wang,
  • Tong Wu,
  • Shentao Zhang,
  • Pingting Guo,
  • Xinyun Lin,
  • Qianyu Lin,
  • Junwen Zhai,
  • Shasha Wu

摘要

Main conclusion

This study overcomes a key technical barrier by establishing transformation and enabling first CRISPR/Cas9 editing in Oxalis, providing a platform for functional genomics and breeding.

Abstract

The lack of an efficient genetic transformation system has considerably hindered functional genomics studies in Oxalis vulcanicola ‘Sunset Velvet’. Here, we established a stable and efficient Agrobacterium-mediated transformation system using stem segments as explants. Key parameters, including pre-culture duration, infection time, Agrobacterium cell density, acetosyringone (AS) concentration, and co-cultivation period, were systematically optimized. Under optimal conditions, the highest transient β-glucuronidase (GUS) expression rate reached approximately 9.0%, and eight stable transgenic lines were successfully obtained. CRISPR/Cas9-mediated genome editing was achieved in Oxalis for the first time. Targeted mutagenesis of OvPDS1, a gene involved in carotenoid biosynthesis, resulted in an albino phenotype, and Sanger sequencing confirmed a base substitution at the target site. Although the editing efficiency was relatively low (0.5%), this result demonstrates the feasibility of genome editing in Oxalis. This study overcomes a major technical bottleneck and provides a robust platform for functional gene analysis, trait improvement, and molecular breeding in O. vulcanicola ‘Sunset Velvet’ and other non-model ornamental plants.