Detection of Furcraea Necrotic Streak Virus (FNSV) in fique seed (Furcraea macrophylla Baker) to prevent the spread of the macana disease

Maira Gamero, D. Ortiz, G. Barrera
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Abstract

The fique crop has great potential for the development of the natural fiber market as a beneficial alternative for the protection of the environment. In Colombia, one of the main limitations of fiber production in fique plants is the viral disease Macana caused by Furcraea Necrotic Streak Virus (FNSV). This work aimed to validate the detection of FNSV in asexual planting material from one of the main producing areas and thus contribute to preventing the spread of the disease in the country. The analysis of plants from different geographic altitudes in Cauca, Colombia, showed a positive correlation with the prevalence of Macana disease (being more significant at higher altitudes) but not with the severity of the symptoms. The detection of FNSV on seeds by dot blot immunobinding assay (DBIA) using a polyclonal antibody IgY showed sensitivity (79 %) and specificity (80 %) when sprouts were analyzed, at the same time, for bulbils, the sensitivity was higher (100 %). Moreover, when sprouts were analyzed by the RT-PCR based on FNSV movement protein and polymerase-associated proteins, the sensitivity and specificity were 94 % and 50 %, respectively, in contrast, in the case of bulbils, the specificity was higher (100 %). Additionally, the results showed no uniformity in the distribution of the viral particles on vegetal tissue of infected plants, which is necessary to use the largest amount of tissue possible to perform the detection. As part of the optimization of the techniques, it was shown that plant tissue samples could be collected, transported, and stored on filter paper, allowing the detection of the virus 60 days after collection.
猕猴桃种子中猕猴桃坏死条纹病毒(FNSV)的检测及防治猕猴桃病的传播
作为一种有益于环境保护的天然纤维替代品,纤维作物具有很大的发展潜力。在哥伦比亚,纤维生产的主要限制之一是由Furcraea Necrotic Streak Virus (FNSV)引起的Macana病毒病。这项工作旨在验证在主要产区之一的无性种植材料中检测到FNSV,从而有助于防止该疾病在该国的传播。对来自哥伦比亚考卡不同地理高度的植物的分析表明,Macana病的流行与Macana病的流行呈正相关(海拔越高越显著),但与症状的严重程度无关。采用IgY多克隆抗体的dot blot免疫结合法(DBIA)检测种子上的FNSV,对芽苗菜的检测灵敏度为79%,特异度为80%,对球茎的检测灵敏度更高(100%)。此外,采用基于FNSV运动蛋白和聚合酶相关蛋白的RT-PCR分析豆芽时,灵敏度和特异性分别为94%和50%,而对于球芽,特异性更高(100%)。此外,结果显示,病毒颗粒在受感染植物的植物组织上分布不均匀,这是使用尽可能多的组织进行检测的必要条件。作为技术优化的一部分,研究表明,植物组织样本可以收集、运输和储存在滤纸上,收集后60天可以检测到病毒。
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