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A method of genetic transformation and gene editing of succulents without tissue culture

文献类型： 外文期刊

作者： Lu, Jinghua 1 ; Li, Shanshan 5 ; Deng, Shuai 6 ; Wang, Mugui 5 ; Wu, Yinghuang 5 ; Li, Ming 5 ; Dong, Jinsong 5 ; Lu, Suhui 6 ; Su, Chunli 6 ; Li, Guofu 6 ; Lang, Zhaobo 3 ; Zhu, Jian-Kang 3 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Sanya Res Inst, Sanya, Peoples R China

2.Hainan Yazhou Bay Seed Lab, Sanya, Peoples R China

3.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen, Peoples R China

4.Southern Univ Sci & Technol, Sch Med, Shenzhen, Peoples R China

5.Chinese Acad Agr Sci CAAS, Natl Nanfan Res Inst, Inst Crop Sci, Sanya, Peoples R China

6.Shandong Shunfeng Biotechnol Co Ltd, Jinan, Peoples R China

7.Minist Agr & Rural Affairs, Key Lab Gene Editing Technol Hainan, Sanya, Peoples R China

8.Chinese Acad Agr Sci CAAS, Ctr Adv Bioind Technol, Beijing, Peoples R China

关键词： cut-dip-budding (CDB) delivery system; gene editing; genetic transformation; succulents

期刊名称：PLANT BIOTECHNOLOGY JOURNAL （ 影响因子：13.8； 五年影响因子：13.2 ）

ISSN： 1467-7644

年卷期： 2024 年

页码：

收录情况： SCI

摘要： Succulents, valued for their drought tolerance and ornamental appeal, are important in the floriculture market. However, only a handful of succulent species can be genetically transformed, making it difficult to improve these plants through genetic engineering. In this study, we adapted the recently developed cut-dip-budding (CDB) gene delivery system to transform three previously recalcitrant succulent varieties - the dicotyledonous Kalanchoe blossfeldiana and Crassula arborescens and the monocotyledonous Sansevieria trifasciata. Capitalizing on the robust ability of cut leaves to regenerate shoots, these plants were successfully transformed by directly infecting cut leaf segments with the Agrobacterium rhizogenes strain K599. The transformation efficiencies were approximately 74%, 5% and 3.9%-7.8%, respectively, for K. blossfeldiana and C. arborescens and S. trifasciata. Using this modified CDB method to deliver the CRISPR/Cas9 construct, gene editing efficiency in K. blossfeldiana at the PDS locus was approximately 70%. Our findings suggest that succulents with shoot regeneration ability from cut leaves can be genetically transformed using the CDB method, thus opening up an avenue for genetic engineering of these plants.

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