Efficient generation of transgenic barley: The way forward to modulate plant-microbe interactions
Efficient generation of transgenic barley: The way forward to modulate plant-microbe interactions
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DOI：
10.1016/j.jplph.2007.06.015
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发表时间：
2008-01-01
影响因子：
4.3
通讯作者：
Kumlehn, Jochen
中科院分区：
文献类型：
--
作者：
Hensel, Goetz;Valkov, Vladimir;Kumlehn, Jochen
稳定的遗传转化是详细阐明植物基因功能的金标准方法。大麦是一种重要的实验模型，广泛应用于应用分子、遗传和细胞生物学研究以及生物技术。介绍了农杆菌介导的基因转移到未成熟胚胎的方案的建立细节，这使得转基因大麦的高效产生。通过对不同外植体处理和共培养条件影响的对比试验，取得了一定的进展。对具有代表性的转基因品系的分析表明，获得的T-DNA拷贝数通常较低，重组DNA的生殖传递符合孟德尔规则，绝大多数初级转基因产生表达各自转基因产物的后代。此外，新建立的协议被证明是有用的，不仅转化高适应性的品种(cv。“金色希望”以及其他春大麦和冬大麦基因型，尽管效率要低得多。作为这项研究的主要结果，一个非常有用的工具，现在可以用于未来的功能基因分析以及基因工程方法。为了修饰大麦中被认为与植物-真菌相互作用有关的基因的表达，利用不同的表达盒已经产生了许多转基因植物。这些植物代表了转化技术如何有助于我们进一步了解重要的生物过程的一个例子。(C) 2007 Elsevier GmbH版权所有。
Stable genetic transformation represents the gold standard approach to the detailed elucidation of plant gene functions. This is particularly relevant in barley, an important experimental model widely employed in applied molecular, genetic and cell biological research, and biotechnology. Presented are details of the establishment of a protocol for Agrobacterium-mediated gene transfer to immature embryos, which enables the highly efficient generation of transgenic barley. Advancements were achieved through comparative experiments on the influence of various explant treatments and co-cultivation conditions. The analysis of representative numbers of transgenic lines revealed that the obtained T-DNA copy numbers are typically low, the generative transmission of the recombinant DNA is in accordance with the Mendelian rules and the vast majority of the primary transgenics produce progeny that expresses the respective transgene product. Moreover, the newly established protocol turned out to be useful to transform not only the highly amenable cultivar (cv.) 'Golden Promise' but also other spring and winter barley genotypes, albeit with substantially lower efficiency. As a major result of this study, a very useful tool is now available for future functional gene analyses as well as genetic engineering approaches. With the aim to modify the expression of barley genes putatively involved in plant-fungus interactions, numerous transgenic plants have been generated using diverse expression cassettes. These plants represent an example of how transformation technology may contribute to further our understanding of important biological processes. (C) 2007 Elsevier GmbH. All rights reserved.