Phytopathology. 2013 Oct;103(10):1069-76. doi: 10.1094/PHYTO-10-12-0253-R.
In the Napa Valley of California, vineyards of 'Cabernet Franc' (CF) clone 214, 'Cabernet Sauvignon' clone 337, and 'Zinfandel' clone 1A (Z1A) with grapevines exhibiting foliar symptoms of red blotches, marginal reddening, and red veins that were accompanied by reduced sugar accumulation in fruit at harvest were initially suspected to be infected with leafroll-associated viruses. However, reverse-transcription polymerase chain reaction (PCR) tests were negative for all known leafroll-associated viruses, with the exception of Grapevine leafroll-associated virus 2 in Z1A. Metagenomic analysis of cDNA libraries obtained from double-stranded RNA enriched nucleic acid (NA) preparations from bark scrapings of dormant canes on an Illumina platform revealed sequences having a distant relationship with members of the family Geminiviridae. Sequencing of products obtained by PCR assays using overlapping primers and rolling circle amplification (RCA) confirmed the presence of a single circular genome of 3,206 nucleotides which was nearly identical to the genome of a recently reported Grapevine cabernet franc-associated virus found in declining grapevines in New York. We propose to call this virus "Grapevine red blotch-associated virus" (GRBaV) to describe its association with grapevine red blotch disease. Primers specific to GRBaV amplified a product of expected size (557 bp) from NA preparations obtained from petioles of several diseased source vines. Chip bud inoculations successfully transmitted GRBaV to test plants of CF, as confirmed by PCR analysis. This is the first report of a DNA virus associated with red blotch disease of grapevines in California.
在加利福尼亚州的纳帕谷，葡萄园中的“品丽珠（CF）克隆 214”、“赤霞珠（CS）克隆 337”和“仙粉黛（Z1A）克隆 1A”葡萄表现出叶片红斑、边缘变红和红色叶脉等症状，这些症状伴随着果实中糖分积累减少，最初怀疑这些葡萄感染了与叶片卷曲相关的病毒。然而，反转录聚合酶链反应（PCR）检测除 Z1A 中的葡萄卷叶伴随病毒 2 外，对所有已知的与叶片卷曲相关的病毒均呈阴性。Illumina 平台上从休眠葡萄藤树皮刮取物中双链 RNA 富集核酸（NA）制备物获得的 cDNA 文库的宏基因组分析揭示了与 Geminiviridae 家族成员具有远缘关系的序列。使用重叠引物和滚环扩增（RCA）进行 PCR 检测获得的产物测序证实，存在一个 3206 个核苷酸的单链圆形基因组，该基因组与最近在纽约衰退葡萄藤中发现的与葡萄品丽珠相关的病毒基因组几乎完全相同。我们建议将这种病毒称为“葡萄红斑伴随病毒”（GRBaV），以描述其与葡萄红斑病的关系。针对 GRBaV 的特异性引物从几个患病来源葡萄藤的叶柄 NA 制备物中扩增出预期大小（557 bp）的产物。芯片芽接种成功地将 GRBaV 传播到 CF 的测试植物中，这通过 PCR 分析得到了证实。这是加利福尼亚州葡萄藤红斑病与 DNA 病毒相关的首次报道。
