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<p>Marigold (<i>Tagetes erecta</i>) is an annual herbaceous plant, well-known for its ornamental, edible, and medicinal properties, primarily due to the abundant secondary metabolites, especially the carotenoid lutein. However, the lack of gene information and genetic validation restricts in-depth understanding of this multiple-benefits horticultural plant. In this study, by targeting the gene of <i>TePDS</i> (<i>phytoene desaturase</i>), we explored the first successful application of the CRISPR/Cas9 system for genome editing in <i>T. erecta</i>. Three guide RNAs (gRNAs) were designed to target distinct exons of <i>TePDS</i>, and CRISPR/Cas9 constructs (p201N-TePDS-1, p201N-TePDS-2, p201N-TePDS-3) were introduced into marigold cultivar ‘Milestone Yellow’ via <i>Agrobacterium</i>-mediated transformation. The obtained sixty-seven transgenic plants indicated transformation efficiency of 67.0%, and 47 of 67 plants (70.1%) showed albino phenotype, including one fully albino transformed marigold and multiple chimeric plants. Eight transgenic plants with varying degrees of albinism and reduced chlorophyll contents were analyzed by sequencing to identify <i>TePDS</i> targeted mutants. Molecular analysis confirmed deletion, insertion and diverse substitutions (a total of 15 editing events), which disrupted <i>TePDS</i> function and led to impaired photosynthesis. Taken together, this work established an efficient CRISPR/Cas9 platform in <i>T</i>. <i>erecta</i>, providing a fundamental tool for secondary metabolism dissection and genetic improvement of this versatile horticultural crop.</p>

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Establishment of an efficient CRISPR/Cas9-mediated editing in marigold (Tagetes erecta)

  • 期刊论文

  • 发表日期：

    2026年4月3日

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  • Qichao Liu,
  • Shaohua Yang,
  • Mingzhu Qiu,
  • Shike She,
  • Feilong Chen,
  • Lei Yao,
  • Minggui Hu,
  • Wei Shi,
  • Xiangli Niu,
  • Yaqiong Qiu

摘要

Marigold (Tagetes erecta) is an annual herbaceous plant, well-known for its ornamental, edible, and medicinal properties, primarily due to the abundant secondary metabolites, especially the carotenoid lutein. However, the lack of gene information and genetic validation restricts in-depth understanding of this multiple-benefits horticultural plant. In this study, by targeting the gene of TePDS (phytoene desaturase), we explored the first successful application of the CRISPR/Cas9 system for genome editing in T. erecta. Three guide RNAs (gRNAs) were designed to target distinct exons of TePDS, and CRISPR/Cas9 constructs (p201N-TePDS-1, p201N-TePDS-2, p201N-TePDS-3) were introduced into marigold cultivar ‘Milestone Yellow’ via Agrobacterium-mediated transformation. The obtained sixty-seven transgenic plants indicated transformation efficiency of 67.0%, and 47 of 67 plants (70.1%) showed albino phenotype, including one fully albino transformed marigold and multiple chimeric plants. Eight transgenic plants with varying degrees of albinism and reduced chlorophyll contents were analyzed by sequencing to identify TePDS targeted mutants. Molecular analysis confirmed deletion, insertion and diverse substitutions (a total of 15 editing events), which disrupted TePDS function and led to impaired photosynthesis. Taken together, this work established an efficient CRISPR/Cas9 platform in T. erecta, providing a fundamental tool for secondary metabolism dissection and genetic improvement of this versatile horticultural crop.