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Background <p>CRISPR/Cas9 technology has garnered increasing attention for its simplicity and precision in genome editing, making it an indispensable tool for gene function research and crop genetic improvement. However, the inefficiency and time-consuming nature of genetic transformation continue to pose substantial challenges to its widespread application in woody plants.</p> Results <p>In this study, we developed a rapid and efficient <i>Agrobacterium</i>-mediated transformation system using petioles as explants for kiwifruit. Positive resistant seedlings were obtained within three months by inoculating on MS medium supplemented with 2.0 mg·L<sup>−1</sup> 6-benzylaminopurine (6-BA), 0.2 mg·L<sup>−1</sup> naphthaleneacetic acid (NAA), and 10 mg·L<sup>−1</sup> hygromycin, which was faster than using leaves as explants. Using this system, CRISPR/Cas9-mediated editing of phytoene desaturase (<i>AcPDS</i>) and <i>ζ</i>-carotene desaturase (<i>AcZDS</i>) achieved an editing efficiency of 20%. Transgenic kiwifruit lines with edited <i>AcZDS</i> exhibited a significant reduction in carotenoid content.</p> Conclusions <p>Overall, we established an efficient <i>Agrobacterium</i>-mediated transformation system using petioles as explants, which is applicable for CRISPR/Cas9-mediated gene editing in kiwifruit, thereby facilitating functional gene studies and genetic improvement.</p>

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CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月9日

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  • Xiaoyan Luo,
  • Ying Dou,
  • Yuxuan Lang,
  • Huamei Zhao,
  • Xinling Liu,
  • Xiaoli Zhang,
  • Yuxing Li,
  • Dong Liang,
  • Hui Xia

摘要

Background

CRISPR/Cas9 technology has garnered increasing attention for its simplicity and precision in genome editing, making it an indispensable tool for gene function research and crop genetic improvement. However, the inefficiency and time-consuming nature of genetic transformation continue to pose substantial challenges to its widespread application in woody plants.

Results

In this study, we developed a rapid and efficient Agrobacterium-mediated transformation system using petioles as explants for kiwifruit. Positive resistant seedlings were obtained within three months by inoculating on MS medium supplemented with 2.0 mg·L−1 6-benzylaminopurine (6-BA), 0.2 mg·L−1 naphthaleneacetic acid (NAA), and 10 mg·L−1 hygromycin, which was faster than using leaves as explants. Using this system, CRISPR/Cas9-mediated editing of phytoene desaturase (AcPDS) and ζ-carotene desaturase (AcZDS) achieved an editing efficiency of 20%. Transgenic kiwifruit lines with edited AcZDS exhibited a significant reduction in carotenoid content.

Conclusions

Overall, we established an efficient Agrobacterium-mediated transformation system using petioles as explants, which is applicable for CRISPR/Cas9-mediated gene editing in kiwifruit, thereby facilitating functional gene studies and genetic improvement.