National Agricultural Research Center, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-8666, Japan.
J Biotechnol. 2012 Feb 10;157(3):421-7. doi: 10.1016/j.jbiotec.2011.12.015. Epub 2011 Dec 27.
The nonstructural Pns9 protein of Rice gall dwarf virus (RGDV) accumulates in viroplasm inclusions, which are structures that appear to play an important role in viral morphogenesis and are commonly found in host cells infected by viruses in the family Reoviridae. An RNA interference construct was designed to target the gene for Pns9 of RGDV, namely Trigger_G9. The resultant transgenic plants accumulated short interfering RNAs specific for the construct. All progenies from self-fertilized transgenic plants had strong and heritable resistance to RGDV infection and did not allow the propagation of RGDV. By contrast, our transgenic plants remained susceptible to Rice dwarf virus, another phytoreovirus. There were no significant changes in the morphology of our transgenic plants compared with non-inoculated wild-type rice plants, suggesting that genes critical for the growth of rice plants were unaffected. Our results demonstrate that the resistance to RGDV of our transgenic rice plants is not due to resistance to the vector insects but to specific inhibition of RGDV replication and that the designed trigger sequence is functioning normally. Thus, our strategy to target a gene for viroplasm matrix protein should be applicable to plant viruses that belong to the family Reoviridae.
水稻锯齿叶矮缩病毒(RGDV)的非结构 Pns9 蛋白在病毒质包涵体内积累,这些包涵体似乎在病毒形态发生中起着重要作用,并且在感染呼肠孤病毒科病毒的宿主细胞中常见。设计了一种针对 RGDV Pns9 基因(即 Trigger_G9)的 RNA 干扰构建体。由此产生的转基因植物积累了针对该构建体的短干扰 RNA。自交转基因植物的所有后代都对 RGDV 感染具有强烈且可遗传的抗性,并且不允许 RGDV 的繁殖。相比之下,我们的转基因植物仍然容易受到另一种植物呼肠孤病毒——水稻矮缩病毒的感染。与未接种的野生型水稻植物相比,我们的转基因植物在形态上没有明显变化,这表明对水稻生长至关重要的基因未受影响。我们的结果表明,我们的转基因水稻植物对 RGDV 的抗性不是由于对载体昆虫的抗性,而是由于对 RGDV 复制的特异性抑制,并且设计的触发序列正常发挥作用。因此,我们针对病毒质基质蛋白基因的靶向策略应该适用于属于呼肠孤病毒科的植物病毒。