Detection and quantification of Agrobacterium vitis, causing crown gall in grapevines, from nursery material

葡萄树冠瘤是由葡萄农杆菌引起的，是一种经济上较为严重的病害。 在大陆性气候的葡萄种植区的一种重要疾病，如英国的奥卡纳根山谷 哥伦比亚。在奥卡纳根山谷的许多葡萄园中都发现了这种疾病，而且似乎在 频率这种细菌在受感染的葡萄血管内移动，并在葡萄树干中形成胆汁。 葡萄藤。这会显著降低植物的活力，并可能导致部分或全部藤本植物死亡，特别是在 幼小的葡萄藤。受感染的植物需要更换。然而，一旦受感染的植物在葡萄园中种植， 细菌可以在葡萄园土壤中存活长达两年，随后可以感染新种植的葡萄。它 有人提出，细菌是通过当地葡萄酒的藤条传入奥卡纳根山谷的 葡萄产业从苗圃采购。在将受感染的藤本植物种植到 无病土壤。当地酿酒葡萄业内部对引入这种疾病的担忧日益加剧。 进入目前没有疾病的新葡萄园地点。许多苗圃为奥卡纳根提供葡萄藤 山谷不需要检测他们的葡萄藤是否感染了葡萄曲霉。目前还没有这种酒的化验方法。 葡萄行业检测收到的材料中是否存在葡萄曲霉。我们正在提议开发一种快速的 基于鉴定适合用于数字液滴的葡萄弧菌DNA引物组的分析 聚合酶链式反应检测和定量检测新种植材料中的葡萄弧菌。然后，该方法将 用于索引苗圃的葡萄藤，通常将葡萄藤运送到奥卡纳根酿酒葡萄行业。这将是 向酿酒葡萄行业提供在哪里可以获得无病葡萄的信息，并确保只有 生产高产的健康葡萄将种植在原始葡萄园土壤中。

Crown gall of grapevines is caused by the bacterium Agrobacterium vitis (A. vitis) and is an economically important disease in grape-growing regions with a continental climate, such as the Okanagan valley in British Columbia. The disease has been found in many vineyards in the Okanagan valley and seems to be increasing in frequency. The bacteria move inside infected grapevine vessels and cause gall formation in trunks of grapevines. This reduces plant vigour significantly and may cause partial or complete vine death, especially in young vines. Infected plants need to be replaced. However, once infected plants are grown in a vineyard, the bacteria can survive in the vineyard soil for up to 2 years and can subsequently infect newly planted vines. It has been proposed that the bacteria are introduced to the Okanagan valley through the vines the local wine grape industry purchase from plant nurseries. The disease develops after planting of the infected vines into disease-free soil. There is growing concern within the local wine grape industry about introducing the disease into new vineyard locations that are currently disease-free. Many nurseries that supply vines to the Okanagan valley are not required to test if their vines are infected with A. vitis. There is no assay available for the wine grape industry to test the received material for the presence of A. vitis. We are proposing to develop a rapid assay based on identification of a suitable DNA primer set specific to A. vitis for use in a digital droplet polymerase chain reaction (PCR) to detect and quantify A. vitis in new planting material. The method will then be used to index vines from nurseries typically delivering vines to the Okanagan wine grape industry. This will provide the wine grape industry with information on where to obtain disease-free vines and to ensure that only healthy vines producing high yields will be planted in virgin vineyard soil.