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，可导致严重减产和增加生产成本。除此之外，他们通常很少或根本没有明显的症状。在此之前，至少有一个广谱病毒抗性基因在园艺油菜作物中被鉴定出来。该基因，eIF(Iso)4E，作为蛋白质复合体的一部分，在mRNA翻译过程中共同作用于帽子识别和结合。人们认为，马铃薯Y病毒科能够与这个复杂的秩序相互作用，利用宿主的细胞机械来感染植物。人们认为，该基因的突变副本能够产生对病毒的抵抗力，因为在帽子结合过程中，它不再能够识别与病毒RNA基因组结合的病毒蛋白。已经证明，在甘蓝基因组中存在eIF4E和eIF(Iso)4E的多个拷贝，然而新的获得完整基因组序列的途径将允许探索是否存在更多的拷贝。此外，编码该复合体其他蛋白的eIF(Iso)4G1和eIF(Iso)4G2的双重突变体也显示出对TuMV的抗性(Nicaise等人，2007年)。该项目旨在探索该复合体的其他成员是否可以产生抗性，以及更好地理解已经识别的基因。此外，利用学到的信息在油菜中创建CRISPR基因敲除，赋予对马铃薯Y病毒科的广谱抗性。

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.