Himmelbach Axel, Zierold Uwe, Hensel Götz, Riechen Jan, Douchkov Dimitar, Schweizer Patrick, Kumlehn Jochen
Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany.
Plant Physiol. 2007 Dec;145(4):1192-200. doi: 10.1104/pp.107.111575. Epub 2007 Nov 2.
Genetic transformation of crop plants offers the possibility of testing hypotheses about the function of individual genes as well as the exploitation of transgenes for targeted trait improvement. However, in most cereals, this option has long been compromised by tedious and low-efficiency transformation protocols, as well as by the lack of versatile vector systems. After having adopted and further improved the protocols for Agrobacterium-mediated stable transformation of barley (Hordeum vulgare) and wheat (Triticum aestivum), we now present a versatile set of binary vectors for transgene overexpression, as well as for gene silencing by double-stranded RNA interference. The vector set is offered with a series of functionally validated promoters and allows for rapid integration of the desired genes or gene fragments by GATEWAY-based recombination. Additional in-built flexibility lies in the choice of plant selectable markers, cassette orientation, and simple integration of further promoters to drive specific expression of genes of interest. Functionality of the cereal vector set has been demonstrated by transient as well as stable transformation experiments for transgene overexpression, as well as for targeted gene silencing in barley.
作物的遗传转化为检验关于单个基因功能的假设以及利用转基因进行目标性状改良提供了可能性。然而，在大多数谷类作物中，长期以来，繁琐且低效的转化方案以及缺乏通用载体系统一直阻碍了这种方法的应用。在采用并进一步改进了农杆菌介导的大麦（Hordeum vulgare）和小麦（Triticum aestivum）稳定转化方案后，我们现在展示了一套通用的二元载体，用于转基因过表达以及通过双链RNA干扰进行基因沉默。该载体集配备了一系列经过功能验证的启动子，并允许通过基于GATEWAY的重组快速整合所需基因或基因片段。额外的内在灵活性体现在植物选择标记的选择、盒式结构的方向以及进一步启动子的简单整合以驱动目标基因的特异性表达。通过瞬时和稳定转化实验，已证明该谷类载体集在转基因过表达以及大麦中靶向基因沉默方面的功能。
