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<p>Sweet pea (<i>Lathyrus odoratus</i>), an important ornamental plant, has been hindered in functional genomics and genetic breeding due to the lack of an efficient transformation and genome editing system. In this study, we developed a simple and efficient CRISPR/Cas9 gene-editing system in sweet pea using an <i>Agrobacterium rhizogenes</i>-mediated (ARM) transformation approach. We constructed a sweet pea genome editing vector, LoGE1, which incorporates a CRISPR/Cas9 system with AtU6-driven sgRNA and CaMV35S-driven Cas9 expression, along with a green fluorescent protein (GFP) reporter. The constructs LoGE1-PDS and LoGE1-SD1 were designed to target <i>LoPDS</i> and <i>LoSD1</i>, respectively, and were introduced into sweet pea using <i>Agrobacterium rhizogenes</i> K599-mediated transformation. Transformation efficiency was evaluated through GFP fluorescence, achieving 54% for LoGE1-PDS and 71% for LoGE1-SD1. Genome editing efficiency at the target sites was 71% for <i>LoPDS</i> and 60% for <i>LoSD1</i>, with various mutation patterns observed. This study provides a foundation for gene editing in sweet pea and offers a reference for developing similar systems in other ornamental plants.</p>

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A simple and efficient Agrobacterium rhizogenes-mediated CRISPR/Cas9 gene editing system in Lathyrus odoratus

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月17日

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  • Yafen Fu,
  • Ximan He,
  • Wenwen Wang,
  • Zhou Chen,
  • Zheng Hong,
  • Bingzhen Du,
  • Chong Wang,
  • Hui Zhang

摘要

Sweet pea (Lathyrus odoratus), an important ornamental plant, has been hindered in functional genomics and genetic breeding due to the lack of an efficient transformation and genome editing system. In this study, we developed a simple and efficient CRISPR/Cas9 gene-editing system in sweet pea using an Agrobacterium rhizogenes-mediated (ARM) transformation approach. We constructed a sweet pea genome editing vector, LoGE1, which incorporates a CRISPR/Cas9 system with AtU6-driven sgRNA and CaMV35S-driven Cas9 expression, along with a green fluorescent protein (GFP) reporter. The constructs LoGE1-PDS and LoGE1-SD1 were designed to target LoPDS and LoSD1, respectively, and were introduced into sweet pea using Agrobacterium rhizogenes K599-mediated transformation. Transformation efficiency was evaluated through GFP fluorescence, achieving 54% for LoGE1-PDS and 71% for LoGE1-SD1. Genome editing efficiency at the target sites was 71% for LoPDS and 60% for LoSD1, with various mutation patterns observed. This study provides a foundation for gene editing in sweet pea and offers a reference for developing similar systems in other ornamental plants.