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在Solanaceous作物中开发病毒诱导的基因组编辑方法

Seo-Young Lee¹, Bomi Kang², Jelli Venkatesh¹

¹Department of Agriculture, Forestry and Bioresources, Research Institute of Agriculture and Life Sciences, Plant Genomics Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea.

Horticulture research

|January 15, 2024

概括

病毒诱导的基因组编辑 (VIGE) 为创建基因组编辑 (GE) 植物的传统方法提供了更快的替代方案. 这项研究开发了使用烟草虫病毒 (TRV) 和土豆病毒X (PVX) 载体的VIGE系统,用于Solanaceous作物,实现显著的突变率.

科学领域:

植物科学植物科学
分子生物学分子生物学
农业生物技术农业生物技术

背景情况:

克里斯普尔/卡斯基因组编辑 (GE) 彻底改变了植物突变发生,但传统方法需要耗时的转基因线.
病毒诱导的基因组编辑 (VIGE) 在模型植物中是有效的,但在Solanaceous作物中未被探索.

研究的目的:

开发和优化VIGE方法用于Solanaceous植物 (西红,马,茄子).
建立转基因介导和无转基因的VIGE方法.
通过热处理来提高VIGE的效率.

主要方法:

在表达Cas9.9的转基因番茄中开发出基于TRV的VIGE.
创建基于PVX的VIGE,用于无转基因的基因组编辑.
设计了针对植物脱和酶的sgRNA,并优化了VIGE条件.
应用热处理 (37°C) 以提高编辑效率.
使用深度测序来评估突变率.
通过组织培养再生编辑植物.

主要成果:

在茄中实现了40.3% (TRV) 和36.5% (PVX) 的突变率,针对Phytoene不和酶.
热处理显著提高了编辑效率: 33-46% (TRV) 和 56-76% (PVX).
成功地将PVX介导的VIGE应用于土豆和茄子.

结论:

开发了基于TRV和PVX的高效VIGE系统,用于Solanaceous植物.
通过PVX介导的VIGE提供了一个无转基因的基因组编辑方法.
通过热处理增强的VIGE方法,为改善作物提供了快速有效的工具.

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