Mendeleum - Institute of Genetics, Faculty of Horticulture, Mendel University in Brno, Valticka 334, Lednice, 691 44, Czech Republic.
Crop Research Institute, Drnovska 507, Praha 6, Ruzyne, 161 06, Czech Republic.
J Virol Methods. 2019 May;267:66-70. doi: 10.1016/j.jviromet.2019.03.003. Epub 2019 Mar 6.
This study describes the application of high-throughput sequencing of small RNA analysis of the efficacy of using Ribavirin to eliminate Grapevine leafroll-associated virus 1, Grapevine fleck virus and Grapevine rupestris stem pitting-associated virus from Vitis vinifera cv. Riesling. The original plant used for sanitation by Ribavirin treatment was one naturally infected with all the viruses mentioned above as confirmed by RT-PCR. A tissue cultures of the plant were established and plantlets obtained were sanitized using Ribavirin. Three years after sanitation, a small RNA sequencing method for virus detection, targeting 21, 22 and 24 nt-long viral small RNAs (vsRNAs), was used to analyze both the mother plant and the sanitized plants. The results showed that the mother plant was infected by the three mentioned viruses and additionally by two viroids - Hop stunt viroid and Grapevine yellow speckle viroid 1. After Ribavirin treatment, the plants contained only the two viroids, with the complete elimination of all the viruses previously present.
本研究描述了高通量测序的小 RNA 分析在使用利巴韦林消除葡萄卷叶相关病毒 1、葡萄斑点病毒和葡萄根瘤蚜相关病毒 1 方面的应用，这些病毒均存在于葡萄品种雷司令中。利巴韦林处理的原始植物是一种自然感染上述所有病毒的植物，这是通过 RT-PCR 确认的。建立了植物的组织培养，并使用利巴韦林对获得的植物进行消毒。消毒 3 年后，使用病毒检测的小 RNA 测序方法，针对 21、22 和 24 个核苷酸长的病毒小 RNA（vsRNA），分析了母株和消毒植物。结果表明，母株受到三种病毒和两种类病毒——Hop stunt viroid 和 Grapevine yellow speckle viroid 1 的感染。经过利巴韦林处理后，植物只含有两种类病毒，先前存在的所有病毒均被完全消除。
