Development of alternative Potyviridae resistance mechanisms through modification of the eIF4E translation initiation complex

植物病毒是作物生产中的一个大问题，在采用杀虫剂等预防措施时，可能导致产量严重下降，并增加生产成本。除此之外，他们通常表现出很少或没有明显的症状。以前在园艺油菜作物中至少鉴定出一种对TuMV （potyvirus dae）具有广谱病毒抗性的基因。这个名为eIF(iso)4E的基因在mRNA翻译过程中，作为一种蛋白质复合物的一部分，在帽识别和结合过程中协同工作。据认为，potyvirus科能够与这个复杂的目相互作用，利用宿主的细胞机制并感染植物。人们认为，该基因的突变副本能够赋予病毒抵抗力，因为它在帽结合过程中不再能够识别与病毒RNA基因组结合的病毒蛋白。已有研究表明，芸芥基因组中存在多个eIF4E拷贝以及同种异构体（eIF(iso)4E），然而，对完整基因组序列的新获取将允许探索是否存在更多拷贝。除此之外，编码该复合体其他蛋白质的eIF(iso)4G1和eIF(iso)4G2双突变体也显示出对TuMV的抗性（Nicaise等，2007）。该项目旨在探索该复合体的其他成员是否可以赋予抗性，以及更好地了解已经确定的基因。此外，利用学到的信息在芸苔中创建CRISPR敲除，赋予对potyvirus科的广谱抗性。

Plant viruses are a large problem in crop production and can cause severe reduction in yield as well as increases in production costs when employing preventative measures such as pesticides. In addition to this they often show little or no visible symptoms. Previously at least one gene conferring broad-spectrum virus resistance to TuMV (Potyviridae) within horticultural brassica crops was identified. The gene, eIF(iso)4E, functions as part of a complex of proteins that work together in the cap recognition and binding during mRNA translation. It is thought that the Potyviridae is able to interact with this complex order to utilise the host's cellular machinery and infect the plant. It is thought that a mutated copy of the gene is able to confer resistance to the virus, as it is no longer able to recognise the viral protein bound to the viral RNA genome during cap binding. It has been shown that there are multiple copies of eIF4E as well as isoform (eIF(iso)4E) within the brassica genome, however new access to a complete genome sequence will allow for the exploration to find if more copies are present. In addition to this, double mutants of eIF(iso)4G1 and eIF(iso)4G2, which code for other proteins from this complex have also showed resistance to TuMV (Nicaise et al., 2007). This project aims to explore weather other members of the complex can confer resistance as well as to better understand the already identified genes. In addition use the information learned to create CRISPR knockouts in Brassica, conferring broad-spectrum resistance to Potyviridae.