在本氏烟植株中检测类病毒 RNA 和 vd-siRNA：类病毒和 vd-siRNA 的 Northern 印迹分析。
Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs.
机构信息
Biology Department, University of Crete, Heraklion, Greece.
IMBB/FoRTH, Heraklion, Greece.
出版信息
Methods Mol Biol. 2022;2316:287-312. doi: 10.1007/978-1-0716-1464-8_24.
Viroids are considered the most minimalistic group of pathogens. Despite their presumed inability to encode for proteins, viroids induce several diseases in plants of primary economic importance. The production of viroid derived siRNAs (vd-siRNAs) of 21-24 nt, accompanies viroid infections in plants and results from the activation of the RNA silencing mechanism and specifically the function of Dicer endonucleases. A comprehensive set of experiments for the study and thorough analysis of viroid-infected plants has been developed. Here we present a detailed experimental plan including optimized protocols for plant infection by agroinfiltration, RNA extraction, and northern blot hybridization for the detection of both viroid genomic RNA and vd-siRNAs.
类病毒被认为是病原体中最简化的群体。尽管它们被认为不能编码蛋白质，但类病毒会在植物中引起几种具有主要经济重要性的疾病。21-24nt 的类病毒衍生 siRNA（vd-siRNA）的产生伴随着植物中的类病毒感染，这是由 RNA 沉默机制的激活引起的，特别是 Dicer 内切酶的功能。已经开发了一套全面的实验方案来研究和彻底分析感染类病毒的植物。在这里，我们提出了一个详细的实验计划，包括通过农杆菌浸润进行植物感染、RNA 提取和 northern blot 杂交的优化方案，以检测类病毒基因组 RNA 和 vd-siRNA。
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