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通过改进的三元载体系统和辅性菌株来提高农菌媒介植物转化效率

Ephraim Aliu^1,2,3, Qing Ji², Anna Wlazlo^1,4

¹Department of Agronomy, Iowa State University, Ames, IA, United States.

Frontiers in plant science

|August 7, 2024

概括

我们开发了新的提米丁辅助性Agrobacterium菌株和三元辅助等离子体,以显著提高Agrobacterium介导的植物转化效率,以实现功能基因组学和作物改进.

科学领域:

植物生物技术植物生物技术
微生物遗传学微生物遗传学
分子生物学分子生物学

背景情况:

在植物功能基因组学和作物改进方面,农业细菌介导的转化是至关重要的.
三级载体系统通过结合额外的毒性 (病毒) 基因来提高T-DNA传递效率.
辅助性Agrobacterium菌株有助于减少培养后的细菌过度生长.

研究的目的:

开发一种新型的提米丁辅助性Agrobacterium菌株和一种新的三元辅助性等离子体.
为了提高Agrobacterium介导的植物转化效率.
为了促进植物转化和基因组编辑应用.

主要方法:

从公开的Agrobacterium菌株 (EHA101,EHA105,EHA105D,LBA4404) 中生成的提米丁辅性菌株.
在Arabidopsis中使用过渡性GUS表达试验评估T-DNA转移能力.
评估了玉米未成熟胚胎中的转化频率,使用了辅性菌株和新的三元辅助体 (pKL2299A).

主要成果:

辅助性菌株表现出依赖蒂米丁的生长,并保留了T-DNA转移能力.
辅助性菌株EHA105Thy-和LBA4404T1的转化频率与玉米中的对照相似.
新的三元辅助器pKL2299A持续改善了玉米的转换频率 (33.3%与25.6%相比).

结论:

开发的辅助性Agrobacterium菌株和三元辅助系统有效地增强了植物的转变.
这种改进的系统为推进植物生物技术和基因组编辑研究提供了有价值的工具.
辅助性菌株简化了Agrobacterium的去除,简化了转化过程.

关键词:

整合系统的整合.基交换突变发生的突变发生.同类重组 (HR) 的方法.玉米的加工加工方法三元向量系统的三元向量系统.

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