Targeting specific genes for RNA interference is crucial to the development of strong resistance to Rice stripe virus
Targeting specific genes for RNA interference is crucial to the development of strong resistance to Rice stripe virus
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DOI：
10.1111/j.1467-7652.2010.00571.x
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发表时间：
2011-05-01
影响因子：
13.8
通讯作者：
Omura, Toshihiro
中科院分区：
文献类型：
--
作者：
Shimizu, Takumi;Nakazono-Nagaoka, Eiko;Omura, Toshihiro
水稻条纹病毒（RSV）在东亚温带地区对水稻生产有严重的负面影响。针对RNA干扰（RNAi）靶序列选择的假定重要性，我们分析了RSV基因组中每个编码基因中潜在靶序列的影响，使用表达一组反向重复序列（IR）构建体的转基因水稻植株。将接种的转基因T-1植物对RSV的反应主观地分为三类，即对RSV的高度抗性、中度抗性和缺乏增强的抗性，即使携带任何构建体的植物在接种RSV之前积累了转基因特异性siRNA。含有对编码核衣壳蛋白的pC 3和编码病毒移动蛋白的pC 4的基因特异的IR构建体的转基因植物对RSV感染具有免疫力，并且比已经用于在田间控制疾病的天然抗性品种对感染具有更高的抗性。相比之下，对编码未知功能的糖蛋白的pC 2基因和编码未知功能的主要非结构蛋白的p4基因特异的IR构建体没有导致抗性。我们的研究结果表明，并非所有针对病毒RNA的RNAi构建体在预防RSV感染方面都同样有效，并且在植物工程中鉴定RNAi攻击的病毒“阿喀琉斯之踵”是重要的。
P>Rice stripe virus (RSV) has a serious negative effect on rice production in temperate regions of East Asia. Focusing on the putative importance of the selection of target sequences for RNA interference (RNAi), we analysed the effects of potential target sequences in each of the coding genes in the RSV genome, using transgenic rice plants that expressed a set of inverted-repeat (IR) constructs. The reactions of inoculated transgenic T-1 plants to RSV were divided subjectively into three classes, namely highly resistant, moderately resistant and lacking enhanced resistance to RSV, even though plants that harboured any constructs accumulated transgene-specific siRNAs prior to inoculation with RSV. Transgenic plants that harboured IR constructs specific for the gene for pC3, which encodes nucleocapsid protein, and for pC4, which encodes a viral movement protein, were immune to infection by RSV and were more resistant to infection than the natural resistant cultivars that have been used to control the disease in the field. By contrast, the IR construct specific for the gene for pC2, which encodes a glycoprotein of unknown function, and for p4, which encodes a major non-structural protein of unknown function, did not result in resistance. Our results indicate that not all RNAi constructs against viral RNAs are equally effective in preventing RSV infection and that it is important to identify the viral 'Achilles heel' for RNAi attack in the engineering of plants.