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

Abstract 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 cont...

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Main Authors: Xiaoyan Luo, Ying Dou, Yuxuan Lang, Huamei Zhao, Xinling Liu, Xiaoli Zhang, Yuxing Li, Dong Liang, Hui Xia
Format: Article
Language:English
Published: BMC 2025-08-01
Series:BMC Plant Biology
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Online Access:https://doi.org/10.1186/s12870-025-07112-6
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author Xiaoyan Luo
Ying Dou
Yuxuan Lang
Huamei Zhao
Xinling Liu
Xiaoli Zhang
Yuxing Li
Dong Liang
Hui Xia
author_facet Xiaoyan Luo
Ying Dou
Yuxuan Lang
Huamei Zhao
Xinling Liu
Xiaoli Zhang
Yuxing Li
Dong Liang
Hui Xia
author_sort Xiaoyan Luo
collection DOAJ
description Abstract 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.
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publishDate 2025-08-01
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spelling doaj-art-95fd5c0e2451435f9fcadafd45e3d7fd2025-08-20T03:04:37ZengBMCBMC Plant Biology1471-22292025-08-0125111310.1186/s12870-025-07112-6CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruitXiaoyan Luo0Ying Dou1Yuxuan Lang2Huamei Zhao3Xinling Liu4Xiaoli Zhang5Yuxing Li6Dong Liang7Hui Xia8College of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityCollege of Horticulture, Sichuan Agricultural UniversityAbstract 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.https://doi.org/10.1186/s12870-025-07112-6CRISPR/Cas9Genetic transformationKiwifruitPhytoene desaturase (PDS)ζ-carotene desaturase (ZDS)
spellingShingle Xiaoyan Luo
Ying Dou
Yuxuan Lang
Huamei Zhao
Xinling Liu
Xiaoli Zhang
Yuxing Li
Dong Liang
Hui Xia
CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
BMC Plant Biology
CRISPR/Cas9
Genetic transformation
Kiwifruit
Phytoene desaturase (PDS)
ζ-carotene desaturase (ZDS)
title CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
title_full CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
title_fullStr CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
title_full_unstemmed CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
title_short CRISPR/Cas9-mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
title_sort crispr cas9 mediated editing of carotenoid biosynthesis genes alters carotenoid concentrations in kiwifruit
topic CRISPR/Cas9
Genetic transformation
Kiwifruit
Phytoene desaturase (PDS)
ζ-carotene desaturase (ZDS)
url https://doi.org/10.1186/s12870-025-07112-6
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