Department of Biological Sciences, University of Lethbridge, Lethbridge, Alberta, Canada T1K 3M4.
Plant Physiol. 2012 Apr;158(4):1503-13. doi: 10.1104/pp.111.192856. Epub 2012 Jan 30.
Genetic transformation of monocotyledonous plants still presents a challenge for plant biologists and biotechnologists because monocots are difficult to transform with Agrobacterium tumefaciens, whereas other transgenesis methods, such as gold particle-mediated transformation, result in poor transgene expression because of integration of truncated DNA molecules. We developed a method of transgene delivery into monocots. This method relies on the use of an in vitro-prepared nano-complex consisting of transferred DNA, virulence protein D2, and recombination protein A delivered to triticale microspores with the help of a Tat2 cell-penetrating peptide. We showed that this approach allowed for single transgene copy integration events and prevented degradation of delivered DNA, thus leading to the integration of intact copies of the transgene into the genome of triticale plants. This resulted in transgene expression in all transgenic plants regenerated from microspores transfected with the full transferred DNA/protein complex. This approach can easily substitute the bombardment technique currently used for monocots and will be highly valuable for plant biology and biotechnology.
单子叶植物的遗传转化仍然对植物生物学家和生物技术人员提出了挑战，因为单子叶植物用农杆菌转化较为困难，而其他转基因方法，如金颗粒介导的转化，由于整合的 DNA 分子是截断的，导致转基因表达较差。我们开发了一种将转基因递送到单子叶植物中的方法。该方法依赖于使用体外制备的纳米复合物，该复合物由转移 DNA、毒性蛋白 D2 和重组蛋白 A 组成，在 Tat2 细胞穿透肽的帮助下递送到黑小麦小孢子中。我们表明，这种方法允许单拷贝转基因整合事件的发生，并防止递送到的 DNA 的降解，从而导致转基因的完整拷贝整合到黑小麦植物的基因组中。这导致了所有用完整转移 DNA/蛋白质复合物转染的小孢子再生的转基因植物中转基因的表达。这种方法可以很容易地替代目前用于单子叶植物的轰击技术，对植物生物学和生物技术具有很高的价值。
