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通过浸花法建立遗传转化和基因编辑方法 于……(原文此处不完整)

Establishment of a Genetic Transformation and Gene Editing Method by Floral Dipping in .

作者信息

Jia Tianjiao, Yang Hua, Zhou Dingding, Zhao Sanzeng, Wang Jianyong, Zhang Tao, Huang Mingkun, Kong Danyu, Liu Yi

机构信息

Lushan Botanical Garden, Jiangxi Province and Chinese Academy of Sciences, NO.9 Zhiqing Road, Jiujiang 332900, China.

出版信息

Plants (Basel). 2024 Oct 10;13(20):2833. doi: 10.3390/plants13202833.

Abstract

L. Webb ex Prantl is used in traditional medicine globally. However, the lack of an efficient and reliable genetic transformation system has seriously limited the investigation of gene function and further utilization of . In this study, a highly efficient, time-saving, and cost-effective -mediated genetic transformation system has been developed in . In this method, the transformation was accomplished by simply dipping developing inflorescences for 45 s into an suspension (OD = 0.6) containing 5% sucrose and 0.03% (/) Silwet L-77. Treated plants were allowed to set seeds which were then plated on a selective medium with hygromycin B (HygB) to screen transformants. Additionally, the CRISPR/Cas9 genomic editing system was validated by targeting phytoene desaturase () gene using this floral dip method, and mutant plants with the expected albino phenotype could be obtained in 2.5 months. This genetic transformation and targeted editing system will be a valuable tool for routine investigation of gene function and further exploitation in .

摘要

L. Webb ex Prantl在全球传统医学中都有应用。然而,缺乏高效可靠的遗传转化系统严重限制了基因功能的研究以及其进一步利用。在本研究中,已在[具体植物名称未给出]中开发出一种高效、省时且经济高效的[转化方法未明确]介导的遗传转化系统。在此方法中,通过将发育中的花序简单地浸入含有5%蔗糖和0.03%(体积分数)Silwet L - 77的[菌液未明确]悬浮液(OD = 0.6)中45秒来完成转化。处理后的植株让其结籽,然后将种子接种在含有潮霉素B(HygB)的选择培养基上筛选转化体。此外,利用这种花浸法靶向八氢番茄红素去饱和酶([基因名称未明确])基因验证了CRISPR/Cas9基因组编辑系统,并且在2.5个月内可获得具有预期白化表型的突变植株。这种遗传转化和靶向编辑系统将成为[具体植物名称未给出]中基因功能常规研究和进一步开发的有价值工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9340/11510603/3305f9678612/plants-13-02833-g001.jpg

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