Accumulated data and results from the recent study of dsRNA isolated from grapevines used in experiments of insect and graft transmission of 'Shiraz' disease

D. Goszczynski
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Abstract

RT-PCR amplicons of dsRNA isolated from various grapevines, which were used in the experiments of transmission of 'Shiraz' disease (SD) from 'Cinsaut Blanc' clone P163/12 to SD-susceptible 'Merlot' and 'Shiraz' using mealybug Planococcus ficus and grafting were investigated. The amplicons were generated in RT-PCR based on virus-specific or random hexamers oligonucleotide primers. Standard molecular techniques and high-throughput sequencing (HTS), respectively, were applied. The results supported the hypothesis that GVA M5v variant present in 'Cinsaut Blanc' P163/12, which is a member of group II of GVA variants associated with SD, is crucial for developing this disease. HTS data did not reveal any other grapevine viruses besides GLRaV-3 and GVA in SD-affected grapevines, except for GVE which, however, was not present in all diseased plants.
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从葡萄中分离的dsRNA用于“设拉子”病昆虫和移植物传播实验的最新研究积累的数据和结果
采用RT-PCR技术,研究了从不同葡萄中分离到的dsRNA的扩增产物,这些葡萄从‘Cinsaut Blanc’无性系P163/12中分离到‘设拉子’病(SD),并利用粉蚧Planococcus ficus和嫁接将SD易感的‘梅洛’和‘设拉子’传播。扩增子是基于病毒特异性或随机六聚体寡核苷酸引物的RT-PCR产生的。分别采用标准分子技术和高通量测序技术。结果支持了GVA M5v变异存在于'Cinsaut Blanc' P163/12中的假设,该变异是与SD相关的GVA变异II组的成员,对该病的发生至关重要。HTS数据未发现除glav -3和GVA外的其他葡萄病毒,但GVE并不存在于所有病株中。
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