College of Resources and Environment, Qingdao Agricultural University, Qingdao, China.
Key Laboratory of Biofuels, Qingdao Engineering Research Center of Biomass Resources and Environment, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.
Plant Biotechnol J. 2019 Nov;17(11):2143-2152. doi: 10.1111/pbi.13128. Epub 2019 May 3.
The fast growth, ease of metabolic labelling and potential for feedstock and biofuels production make duckweeds not only an attractive model system for understanding plant biology, but also a potential future crop. However, current duckweed research is constrained by the lack of efficient genetic manipulation tools. Here, we report a case study on genome editing in a duckweed species, Lemna aequinoctialis, using a fast and efficient transformation and CRISPR/Cas9 tool. By optimizing currently available transformation protocols, we reduced the duration time of Agrobacterium-mediated transformation to 5-6 weeks with a success rate of over 94%. Based on the optimized transformation protocol, we generated 15 (14.3% success rate) biallelic LaPDS mutants that showed albino phenotype using a CRISPR/Cas9 system. Investigations on CRISPR/Cas9-mediated mutation spectrum among mutated L. aequinoctialis showed that most of mutations were short insertions and deletions. This study presents the first example of CRISPR/Cas9-mediated genome editing in duckweeds, which will open new research avenues in using duckweeds for both basic and applied research.
快速生长、易于进行代谢标记以及有生产原料和生物燃料的潜力，使得浮萍不仅成为植物生物学研究的理想模式系统，而且还有望成为未来的一种作物。然而，当前浮萍的研究受到缺乏有效遗传操作工具的限制。在这里，我们报告了利用快速高效的转化和 CRISPR/Cas9 工具对浮萍物种 Lemna aequinoctialis 进行基因组编辑的案例研究。通过优化现有的转化方案，我们将农杆菌介导的转化时间缩短至 5-6 周，成功率超过 94%。基于优化的转化方案，我们利用 CRISPR/Cas9 系统生成了 15 个（成功率为 14.3%）双等位基因 LaPDS 突变体，它们表现出白化表型。对突变的 L. aequinoctialis 中 CRISPR/Cas9 介导的突变谱的研究表明，大多数突变是短插入和缺失。本研究首次展示了在浮萍中利用 CRISPR/Cas9 进行基因组编辑的范例，这将为利用浮萍进行基础和应用研究开辟新的途径。
