Flores-Sandoval Eduardo, Dierschke Tom, Fisher Tom J, Bowman John L
School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia Department of Plant Biology, UC Davis, Davis, CA 95616, USA
Plant Cell Physiol. 2016 Feb;57(2):281-90. doi: 10.1093/pcp/pcv068. Epub 2015 May 13.
We describe the efficient use of artificial microRNAs (amiRs) in Marchantia polymorpha using both endogenous and heterologous primary microRNA (pri-miR) hairpin backbones. Targeting of two transcription factor genes, MpARF1 and MpRR-B, mediating different hormonal responses, demonstrated that amiRs can create specific and reproducible physiological and morphological defects, facilitating interpretation of gene function. A third amiR was designed to target a gene encoding a component of the Polycomb recessive complex 2, MpE(z), and constitutive expression of this amiR results in sporeling lethality. Adaptation of an estrogen-inducible system allowed analysis of the phenotypic effects of induction of this amiR during other stages of the life cycle. We discuss the advantages and challenges of the use of amiRs as a tool for reverse genetic analysis in M. polymorpha.
我们描述了在多歧苏铁中利用内源性和异源初级微小RNA（pri-miR）发夹骨架高效使用人工微小RNA（amiR）的情况。靶向两个介导不同激素反应的转录因子基因MpARF1和MpRR-B，证明amiR可以产生特定且可重复的生理和形态缺陷，便于对基因功能进行解读。设计了第三个amiR来靶向编码多梳隐性复合体2组分MpE(z)的基因，该amiR的组成型表达导致幼苗致死。雌激素诱导系统的适应性改造使得能够分析在生命周期的其他阶段诱导该amiR所产生的表型效应。我们讨论了将amiR用作多歧苏铁反向遗传分析工具的优势和挑战。
