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工程玉米根微生物组:一个快速的MoClo工具包和潜在的细菌底盘的识别研究植物微生物相互作用

John van Schaik¹, Zidan Li¹, John Cheadle¹

¹Department of Chemical and Biomolecular Engineering, North Carolina State University, Room 2109, Partners II, 840 Main Campus Drive, Raleigh, North Carolina 27606, United States.

ACS synthetic biology

|September 15, 2023

概括

研究人员开发了一套玉米根微生物组工具包 (MRMTK),用于设计与植物相关的细菌. 这种工具包能够快速生成遗传结构和高效转换,有助于通过植物微生物相互作用研究和增强作物生产.

科学领域:

微生物学微生物学
合成生物学合成生物学
农业科学农业科学

背景情况:

可持续的作物生产对于全球粮食安全至关重要.
与植物相关的微生物组为提高作物产量提供了潜力.
有限的遗传工具阻碍了对这些有益微生物的研究和工程.

研究的目的:

开发一个多功能基因工程工具包,用于一个模型的玉米根合成社区.
为关键的根共生细菌建立高效的转化协议.
为了实现植物微生物相互作用的阐明和工程,以改善作物生长.

主要方法:

使用模块化克隆 (MoClo) 开发了玉米根微生物组工具包 (MRMTK).
在五种细菌菌株中产生异质基因表达的构造: *Enterobacter ludwigii*, *Pseudomonas putida*, *Herbaspirillum robiniae*, *Stenotrophomonas maltophilia* 和 *Brucella pituitosa*.
优化了高效的转换协议,并通过基因组促进器捕获识别了内源表达序列 (ES).

主要成果:

该MRMTK允许快速 (2,5小时) 生成多样化的遗传结构 (5400个潜在的等离子体).
在所有目标菌株中实现了高转换效率 (10^7-10^9转换剂/μg DNA).
确定了特定菌株的内源性促进体和用于基因表达的核糖体结合部位.

结论:

MRMTK提供了一个可扩展和可适应的平台,用于根共生动物的基因工程.
该工具包和标准化转化方法促进了对植物微生物相互作用的研究.
释放了设计有益的植物微生物关系的潜力,以促进作物生长.

关键词:

电穿孔的电穿孔是一种玉米玉米玉米是一种模块化克隆是一种模块化克隆.活动主办人陷合成社区合成社区工具包是一个工具包.

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