Singh Kashmir, Kaur Ravneet, Qiu Wenping
Center for Grapevine Biotechnology, William H. Darr School of Agriculture, Missouri State University, Mountain Grove, MO, USA.
Methods Mol Biol. 2012;883:177-91. doi: 10.1007/978-1-61779-839-9_14.
Small RNAs (sRNAs) have emerged as one of the most important regulators of gene expression in eukaryotes. sRNAs are intermediate molecules as well as end products in the antiviral defense pathway called RNA interference in plants and animals. Profiling of sRNAs using next-generation sequencing technologies has identified a number of plant viruses that have never been reported previously, and has provided a deeper view of virus populations in a plant that cannot be achieved by conventional methods like PCR and ELISA. In this chapter, we describe the methodology of deep sequencing of sRNAs. The high-throughput and highly sensitive method will revolutionize the identification of plant viruses and the study of molecular plant-virus interactions.
小RNA（sRNA）已成为真核生物中基因表达最重要的调节因子之一。在植物和动物中，sRNA是抗病毒防御途径（称为RNA干扰）中的中间分子以及终产物。利用下一代测序技术对sRNA进行分析，已鉴定出许多以前从未报道过的植物病毒，并提供了对植物中病毒群体的更深入了解，这是传统方法（如PCR和ELISA）无法实现的。在本章中，我们描述了sRNA深度测序的方法。这种高通量且高度灵敏的方法将彻底改变植物病毒的鉴定以及植物-病毒分子相互作用的研究。
