Seasonal dynamics of viral diseases in Ontario vineyards: mapping optimal time window for sensitive and reliable detection of major viruses afflicting the grape/wine industry

病毒是葡萄和葡萄酒的健康、生产力、质量和盈利的主要障碍。 加拿大和世界上许多其他国家的工业。自2013年以来，由病毒引起的疾病一直在 在加拿大主要的葡萄和葡萄酒产地尼亚加拉地区观察和检测到。这场疾病暴发 已经引起了许多葡萄种植者和酿酒厂的重大担忧，因为病毒疾病严重困扰着 优质葡萄及相关葡萄酒产品的生产。此外，病毒感染将延长 对受感染葡萄的健康和可持续性的影响，撕裂的葡萄园最终会失去生产力，因为 疾病的发展和恶化。葡萄和葡萄酒行业在高度发展方面亟需帮助 灵敏可靠的技术，可快速检测所有主要病毒。然而，检测的可靠性 取决于总核酸的质量，要使用的引物的普遍性(谱)，以及 抽样的最佳时间窗口。在接洽计划支持的前两个项目中，我们有 开发了一种非常成功的从不同类型葡萄中分离优质RNA的技术 在整个生长季节收集的组织。我们还设计了广谱引物， 能够检测到每种目标病毒的所有遗传变异。不幸的是，关于最佳时间的信息 在全球范围内，尤其是在安大略省，无法获得检测每一种目标病毒的框架。这个 这项研究项目的目标是揭示安大略省五种目标病毒的季节性动态 葡萄园代表着气候凉爽的葡萄栽培条件。我们将使用的方法包括基于核酸的 技术(RT-PCR和RT-qPCR)和基于抗体的方法(Western Blotting)。这些信息 该项目的结果不仅将使安大略省的葡萄和葡萄酒行业受益，而且还将惠及其他葡萄 加拿大和其他许多国家的产区。因此，本研究将具有深远的经济意义。 对加拿大农业和农业食品部门以及旅游业的影响。

Viruses represent a major impediment to the health, productivity, quality and profitability of the grape and wine industry in Canada and many other countries of the world. Since 2013, diseases caused by viruses have been observed and detected in Niagara region, the major grape and wine producer in Canada. This disease outbreak has caused major concern to the many grape growers and wineries as viral diseases severely afflict the production of quality grapes and the related wine products. Furthermore, viral infections will have prolonged effects to the health and sustainability of the infected vines, rending vineyards eventually lose productivity as diseases progress and worsen. The grape and wine industry is in dire need of help in the development of highly sensitive and reliable technologies for the quick detection of all major viruses. However, reliability of detection depends on the quality of total nucleic acids, the universality (spectrum) of primers to be used, as well as the optimal window of time for sampling. In two previous project supported by the Engage program, we have developed a highly successful technology for the isolation of quality RNAs from different types of grapevine tissue collected throughout the entire growing season. We have also designed broad-spectrum primers that are able to detect all genetic variants of each of the target viruses. Unfortunately, information on the best time frame for the detection of each of these target viruses is not available, globally and especially in Ontario. The goal of this research project is to uncover the seasonal dynamics of each of the five targeted viruses in Ontario vineyards representing cool climate viticulture conditions. The methods we will use include nucleic acid-based technologies (RT-PCR and RT-qPCR) and antibody-based methods (Western blotting). The information resulting from this project will not only benefit the grape and wine industries in Ontario but also other grape production regions of Canada and many other countries. Thus, this research will have far-reaching economic impact on the well being of the agriculture and agri-food sector and tourism industry in Canada.