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通过使用CRISPR/Cas9系统对MLO基因进行向突变,增强大豆中粉状菌的耐药性

Thao Phuong Bui^1,2, Huy Le^1,3, Dong Thi Ta¹

¹Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi, Vietnam.

BMC plant biology

|November 3, 2023

概括

研究人员使用CRISPR/Cas9来制造大豆植物,通过突变GmMLO基因,增强对粉的抵抗力. 这些基因编辑的大豆显示疾病严重程度降低,为改善作物提供了一个有希望的途径.

科学领域:

植物病理学植物病理学
分子遗传学分子遗传学
农业生物技术农业生物技术

背景情况:

粉状菌显著降低了大豆产量和种子质量.
菌局部O (MLO) 基因是植物对粉状菌的关键易感因素.
开发抗病大豆品种对于作物保护至关重要.

研究的目的:

利用CRISPR/Cas9基因编辑来创造具有对粉状菌的增强抵抗力的大豆突变种.
调查大豆GmMLO基因对粉易感受性的作用.

主要方法:

设计并使用双sgRNACRISPR/Cas9构造用于向突变发生.
采用Agrobacterium tumefaciens介导的转化,将构造物引入大豆品种DT26.
通过T1和T2世代确认和验证了CRISPR/Cas9诱导的突变.

主要成果:

产生了各种GmMLO突变形式,包括三重和四重淘汰 (Gmmlo02/Gmmlo19/Gmmlo23和Gmmlo02/Gmmlo19/Gmmlo20/Gmmlo23).
突变的大豆植物表现出对Erysiphe diffusa的增强抵抗力,粉状菌的严重程度降低了高达36.4%.
在突变系中没有观察到对大豆生长或发育的显著类效应.

结论:

GmMLO02,GmMLO19,GmMLO20和GmMLO23基因参与大豆对粉的敏感性.
通过CRISPR/Cas9,成功地产生了无转基因,同胞性突变的大豆菌株,改善了粉状菌的耐药性.
这些基因编辑的大豆种系有可能成为未来的培育计划,旨在提高抗病能力.

关键词:

这就是CRISPR/Cas9的作用.有关GmMLO基因的基因粉状菌的耐药性粉状菌的耐药性豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆豆有针对性的突变.

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