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在Fusarium verticillioides中开发一个高效的CRISPR/Cas9系统及其在减少真菌毒素污染中的应用

Tingting Tang¹, Yi Ding¹, Wei Guo¹

¹Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Beijing 100193, P. R. China.

Journal of agricultural and food chemistry

|May 27, 2024

概括

研究人员开发了一种高效的CRISPR/Cas9基因组编辑系统,用于主要的玉米病原体Fusarium verticillioides. 该系统可实现精确的基因编辑和生物控制剂的制造,以减少农作物中的真菌毒素污染.

科学领域:

植物病理学植物病理学
菌类学菌类学是指菌类学.
分子生物学分子生物学

背景情况:

Fusarium verticillioides 是玉米的重要真菌病原体,导致大量的产量损失和产生有害的真菌毒素.
有限的遗传工具阻碍了F. verticillioides的研究和管理策略.

研究的目的:

建立一个高效的CRISPR/Cas9基因组编辑系统,用于F. verticillioides.
开发用于基因操纵和生物控制这一重要的玉米病原体的工具.

主要方法:

使用具有AMA1序列的自主复制等离子体开发CRISPR/Cas9系统.
通过非同类末端结合 (NHEJ) 和使用双 sgRNA 表达的精确基因删除来破坏基因.
建立一个pyrG循环系统,并产生富蒙尼辛生物合成突变物.

主要成果:

在pyrG和pyrE的基因破坏方面实现了高效率 (66-100%).
成功精确的基因删除和设计者染色体的生成,包括38kb片段删除用于富蒙尼辛生物合成.
开发的 ΔFUM1 和 ΔFUM 突变体证明了它们作为生物控制剂的潜力,可以防止米尼辛 B1 的污染.

结论:

已经建立的CRISPR/Cas9系统显著提高了F. verticillioides的基因操纵能力.
开发的基因组编辑工具和生物控制剂为管理F. verticillioides和减轻对玉米的影响提供了有价值的策略.

关键词:

这就是CRISPR/Cas9的作用.在Fusarium verticillioides中使用.这是一种抗氧化菌株.富莫尼西尼的存在乌拉的辅助性选择系统.

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