Department of Plant Pathology, Irrigated Agriculture Research and Extension Center, Washington State University, Prosser, WA 99350.
Department of Agriculture, Columbia Basin College, Pasco, WA 99301.
Plant Dis. 2018 Nov;102(11):2129-2135. doi: 10.1094/PDIS-12-17-1962-RE. Epub 2018 Sep 17.
Vineyard surveys were conducted for three consecutive seasons in eastern Washington State, the major grapevine-growing region in the state, to document the occurrence of Grapevine leafroll-associated virus 3 (GLRaV-3) and Grapevine red blotch virus (GRBV). The majority of samples were collected from red-berried wine grape (Vitis vinifera) cultivars exhibiting symptoms of or suspected for grapevine leafroll (GLD) and red blotch (GRBD) diseases. A limited number of samples from white-berried cultivars were collected randomly due to the lack of visual symptoms. Samples were collected from a total of 2,063 grapevines from 18 red-berried cultivars and seven white-berried cultivars planted in eight American Viticultural Areas and tested for GLRaV-3 and GRBV using RT-PCR and PCR, respectively. The results showed 67.77% and 6.01% of total samples positive for GLRaV-3 and GRBV, respectively, and 9.06% of samples positive for both viruses. About 17% of samples tested negative for the two viruses, but some of these samples were positive for GLRaV-2 and GLRaV-4. Overall results indicated that GLRaV-3 was more common than GRBV, independent of cultivars and the geographic origin of samples. Due to variability in symptoms in red-berried cultivars, virus-specific diagnostic assays were deemed necessary for reliable identification of GLRaV-3 and GRBV and to differentiate GLD and GRBD symptoms from those induced by biotic and abiotic stresses in vineyards. A multiplex PCR protocol was developed for simultaneous detection of GLRaV-3 and GRBV in grapevine samples. A global phylogenetic analysis of GRBV genome sequences revealed segregation of virus isolates from Washington State vineyards into two distinct clades, with the majority of isolates belonging to clade II.
在华盛顿州东部（该州主要的葡萄种植区）进行了三个连续季节的葡萄园调查，以记录葡萄卷叶伴随病毒 3（GLRaV-3）和葡萄红斑病毒（GRBV）的发生情况。大多数样本是从表现出葡萄卷叶（GLD）和红斑（GRBD）症状或疑似症状的红浆果酿酒葡萄（Vitis vinifera）品种中采集的。由于缺乏视觉症状，仅从白浆果品种中随机采集了有限数量的样本。从 18 个红浆果品种和 7 个白浆果品种的总共 2063 株葡萄藤中采集了样本，这些葡萄藤种植在 8 个美国葡萄栽培区，分别使用 RT-PCR 和 PCR 检测 GLRaV-3 和 GRBV。结果表明，GLRaV-3 和 GRBV 的总阳性样本分别为 67.77%和 6.01%，两种病毒均为阳性的样本为 9.06%。约 17%的样本检测到两种病毒均为阴性，但其中一些样本对 GLRaV-2 和 GLRaV-4 呈阳性。总体结果表明，无论品种和样本的地理来源如何，GLRaV-3 都比 GRBV 更为常见。由于红浆果品种的症状存在差异，因此需要进行病毒特异性诊断检测，以可靠地鉴定 GLRaV-3 和 GRBV，并区分 GLD 和 GRBD 症状与葡萄园中的生物和非生物胁迫引起的症状。开发了一种用于同时检测葡萄藤样本中 GLRaV-3 和 GRBV 的多重 PCR 方案。对 GRBV 基因组序列的全球系统发育分析表明，来自华盛顿州葡萄园的病毒分离物分为两个不同的分支，大多数分离物属于分支 II。
