Evaluation of spray-induced gene silencing as a method to reduce the spread of Blueberry Shock virus in Canadian blueberry farms

加拿大的蓝莓种植者，主要集中在不列颠哥伦比亚省，因蓝莓休克病毒的到来和传播而遭受了越来越多的重大损失。对感染的反应是令人震惊的，因为树叶和花朵在春天出现枯萎，导致在那个季节没有可收获的果实。植物在1-3年内恢复，但由于在这一恢复期产量的损失，种植者付出了巨大的代价。恢复后，植物仍然有病毒，通过蜜蜂和其他传粉媒介的花粉在整个种植过程中和种植之间传播。由于蜜蜂授粉对浆果生产是必要的，所以处理不能影响蜜蜂。另一方面，有效的治疗只需要在开花时施用，以限制病毒通过花粉传播。在这里，我们提议测试一种被称为喷雾诱导基因沉默的新方法的使用。它的原理是向植物喷洒与病毒基因组匹配并结合的双链核糖核酸（dsRNA），从而触发病毒的毁灭。我们将进行温室对照实验，以评估(1)应用RNA是否可以减少受感染植物中的病毒数量，(2)是否可以通过对开花植物施用RNA来防止花粉感染。RNA是存在于所有活细胞和生物体中的一种天然生物物质，具有固有的不稳定性和无毒性。由于所使用的RNA是针对该病毒的，因此该方法不仅有可能提供针对蓝莓休克病毒传播的第一个治疗方法，而且还可能为加拿大农业中未来害虫生物防治的发展提供一个例子。

The blueberry growers of Canada, focused in British Columbia, have experienced increasing and substantial losses from the arrival and spread of the Blueberry Shock Virus. The response to infection is shock-like, as leaves and flowers appear blighted in the spring, resulting in no harvestable fruit in that season. Plants recover in 1-3 years, but there is a substantial cost to the grower due to the loss of yield during this recovery period. After recovery, plants still have the virus, which is spread via pollen by honey bees and other pollinators throughout and between plantings. As pollination from bees is necessary for berry production, treatments can not affect bees. On the other hand, an effective treatment would only need to be applied at the time of flowering to limit the spread of the virus through pollen. Here we propose to test the use of a novel methodology known as Spray-Induced Gene Silencing. It is based on spraying the plant with double-stranded ribonucleic acid (dsRNA) matching and binding to the genome of the virus, triggering its destruction. We will carry out controlled greenhouse experiments to assess (1) if the number of viruses in infected plants can be reduced by RNA application, and (2) if Infection via pollen can be prevented by applying RNA to flowering plants. RNA is a natural biological substance present in all living cells and organisms and is inherently unstable and non-toxic. Since the used RNA is targeted for this virus, this method has the potential of providing not only the first treatment against the spread of Blueberry Shock Virus but also provide an example for the development of future biological control of pests in Canadian agriculture.