Agriculture and Agri-Food Canada, Pacific Agri-Food Research Center, Summerland, British Columbia V0H 1Z0, Canada.
J Genet Genomics. 2013 May 20;40(5):261-70. doi: 10.1016/j.jgg.2013.03.012. Epub 2013 Apr 11.
The utility of artificial microRNAs (amiRNAs) to induce loss of gene function has been reported for many plant species, but expression efficiency of the different amiRNA constructs in different transgenic plants was less predictable. In this study, expressions of amiRNAs through the gene backbone of Arabidopsis miR168a were examined by both Agrobacterium-mediated transient expression and stable plant genetic transformation. A corresponding trend in expression of amiRNAs by the same amiRNA constructs between the transient and the stable expression systems was observed in the experiments. Plant genetic transformation of the constructs that were highly expressible in amiRNAs in the transient agro-infiltration assays resulted in generation of transgenic lines with high level of amiRNAs. This provides a simple method for rapid and effective selection of amiRNA constructs used for a time-consuming genetic transformation in plants.
人工 microRNAs（amiRNAs）在许多植物物种中诱导基因功能丧失的效用已有报道，但不同 amiRNA 构建体在不同转基因植物中的表达效率较难预测。在这项研究中，通过拟南芥 miR168a 的基因骨架，通过农杆菌介导的瞬时表达和稳定植物遗传转化来检测 amiRNAs 的表达。在实验中，相同 amiRNA 构建体在瞬时表达和稳定表达系统中的表达趋势一致。在瞬时农杆菌浸润分析中高表达 amiRNAs 的构建体的植物遗传转化导致产生高水平 amiRNAs 的转基因系。这为快速有效地选择用于植物耗时遗传转化的 amiRNA 构建体提供了一种简单的方法。
