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Updated: Jul 24, 2025

Scalable Transfection of Maize Mesophyll Protoplasts

Scalable Transfection of Maize Mesophyll Protoplasts

Published on: June 23, 2023

玉米中原体的可扩展性转染

Jackson Tonnies¹, Nicholas A Mueth², Sayeh Gorjifard¹

¹Genome Sciences Department, University of Washington.

Journal of visualized experiments : JoVE

|July 10, 2023

概括

这项研究提出了一种简化方法,用于传染数百万种玉米叶 mesophyll 原塑体,改进了以前用于将 DNA 插入植物细胞的技术. 这一进步使得在玉米中进行大规模的基因组实验成为可能.

科学领域:

植物生物学植物生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

玉米中细胞转移通常涉及细胞壁消化形成原生质,随后通过使用电穿孔或聚乙烯糖醇 (PEG) 等方法插入DNA.
现有的方法产生了成千上万的转染原塑体,限制了某些遗传实验的规模.

研究的目的:

开发一种简单且可扩展的方法来隔离和转移数百万种玉米叶中原体.
为了促进玉米的基因组规模实验,例如大规模并行记者测试.

主要方法:

开发了一个精简的原塑化协议,省略了W5洗等步骤.
优化离心,聚乙烯糖醇 (PEG) 介导的转染和化步骤对大量原生质细胞进行了实施.
该方法的重点是从玉米 (Zea mays L.) 中有效分离和转移叶子中原体.

主要成果:

成功地分离和转染了数百万种玉米叶中粒细胞原质体,与以前的方法相比,产量显著增加.
精简的工艺通过修改常见的原塑和转化步骤来提高效率.
该方法支持塑体结构的大型库的表达.

结论:

开发的方法为玉米原质细胞转化提供了一个可扩展的方法.
这种技术对于在玉米中实现高通量,基因组规模的功能基因组学研究至关重要.
增强的原质细胞产量和转染效率为玉米的遗传研究开辟了新的途径.

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Direct Agroinoculation of Maize Seedlings by Injection with Recombinant Foxtail Mosaic Virus and Sugarcane Mosaic Virus Infectious Clones

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