Use of siRNAs for Diagnosis of Viruses Associated to Woody Plants in Nurseries and Stock Collections
Use of siRNAs for Diagnosis of Viruses Associated to Woody Plants in Nurseries and Stock Collections
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DOI：
10.1007/978-1-4939-7683-6_9
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Varallyay, Eva
中科院分区：
文献类型：
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作者：
Czotter, Nikoletta;Molnar, Janos;Varallyay, Eva
多年生木本植物，如葡萄藤和果树，可以被几种病毒感染，甚至是多重感染。由于它们是无性繁殖的，繁殖材料（砧木和品种）的植物检疫状况可能对新种植园的寿命和健康产生深远的影响。测序技术的快速发展为基于宏基因组学的病毒诊断提供了新的机会。在病毒感染期间由宿主免疫系统产生的病毒衍生的小RNA可以通过下一代技术进行测序并分析病毒的存在，从而揭示样品中所有已知病毒病原体的存在。该方法基于Illumina短RNA测序和宿主中病毒衍生的小RNA的生物信息学分析。在这里，我们描述了这种具有挑战性的技术一步一步的协议，以确保每个用户的成功。
Woody perennial plants like grapevine and fruit trees can be infected by several viruses even as multiple infections. Since they are propagated vegetatively, the phytosanitary status of the propagation material (both the rootstock and the variety) can have a profound effect on the lifetime and health of the new plantations. The fast evolution of sequencing techniques provides a new opportunity for metagenomics-based viral diagnostics. Viral derived small RNAs produced by the host immune system during viral infection can be sequenced by next-generation techniques and analyzed for the presence of viruses, revealing the presence of all known viral pathogens in the sample. This method is based on Illumina sequencing of short RNAs and bioinformatics analysis of virus-derived small RNAs in the host. Here we describe a protocol for this challenging technique step by step with notes, in order to ensure success for every user.