Morán Félix, Sassalou Chrysoula-Lito, Canales Celia, Maliogka Varvara I, Olmos Antonio, Ruiz-García Ana Belén
Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera km 4.5, Moncada, 46113 Valencia, Spain.
Plant Pathology Laboratory, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Plants (Basel). 2020 Sep 4;9(9):1151. doi: 10.3390/plants9091151.
Grapevine Roditis leaf discoloration-associated virus (GRLDaV) is an emerging grapevine pathogen included in the European and Mediterranean Plant Protection Organization (EPPO) alert list due to its ability to damage grapevine crops and cause production losses. This work aimed to develop a specific and reliable diagnostic tool that would contribute to preventing the spread of this pathogen. Therefore, a TaqMan real-time quantitative PCR was developed. The method was validated according to EPPO guidelines showing a high degree of analytical sensitivity, analytical specificity, selectivity, and repeatability and reproducibility. The sensitivity of this method is much higher than the sensitivity reached by previously reported methods even when tested in crude extracts, which could allow rapid testing by avoiding nucleic acid extraction steps. The method was also able to detect GRLDaV isolates from all the geographic origins reported so far, despite their high degree of genetic diversity. In addition, this new technique has been successfully applied for the quantitative detection of GRLDaV in plant material and two mealybug species, and . In conclusion, the methodology developed herein represents a significant contribution to the diagnosis and control of this emerging pathogen in grapevine.
葡萄罗迪蒂斯叶变色相关病毒（GRLDaV）是一种新出现的葡萄病原体，因其能够损害葡萄作物并导致产量损失，被列入欧洲和地中海植物保护组织（EPPO）警报清单。这项工作旨在开发一种特异性和可靠的诊断工具，以有助于防止这种病原体的传播。因此，开发了一种TaqMan实时定量PCR方法。该方法根据EPPO指南进行了验证，显示出高度的分析灵敏度、分析特异性、选择性以及重复性和再现性。即使在粗提物中进行测试，该方法的灵敏度也远高于先前报道的方法，这可以通过避免核酸提取步骤实现快速检测。尽管GRLDaV分离株具有高度的遗传多样性，但该方法仍能够检测到迄今为止报道的所有地理来源的分离株。此外，这项新技术已成功应用于植物材料以及两种粉蚧（ 和 ）中GRLDaV的定量检测。总之，本文开发的方法对这种新出现的葡萄病原体的诊断和控制做出了重大贡献。
