基于马铃薯X病毒的马铃薯微小RNA沉默（VbMS）

Potato Virus X-Based microRNA Silencing (VbMS) In Potato.

作者信息

Zhao Jinping, Rios Carlos Garcia, Song Junqi

机构信息

Texas A&M AgriLife Research Center at Dallas, Texas A&M University System.

Texas A&M AgriLife Research Center at Dallas, Texas A&M University System; Department of Plant Pathology, Microbiology, Texas A&M University;

出版信息

J Vis Exp. 2020 May 11(159). doi: 10.3791/61067.

DOI:10.3791/61067

PMID:32449744

Abstract

Virus-based microRNA silencing (VbMS) is a rapid and efficient tool for functional characterization of microRNAs (miRNAs) in plants. The VbMS system has been developed and applied for various plant species including Nicotiana benthamiana, tomato, Arabidopsis, cotton, and monocot plants such as wheat and maize. Here, we describe a detailed protocol using PVX-based VbMS vectors to silence endogenous miRNAs in potato. To knock down the expression of a specific miRNA, target mimic (TM) molecules of miRNA of interest are designed, integrated into plant virus vectors, and expressed in potato by Agrobacterium infiltration to bind directly to the endogenous miRNA of interest and block its function.

摘要

基于病毒的 microRNA 沉默（VbMS）是一种用于植物中 microRNA（miRNA）功能表征的快速且高效的工具。VbMS 系统已被开发并应用于多种植物物种，包括本氏烟草、番茄、拟南芥、棉花以及单子叶植物如小麦和玉米。在此，我们描述了一种使用基于 PVX 的 VbMS 载体在马铃薯中沉默内源性 miRNA 的详细方案。为了敲低特定 miRNA 的表达，设计感兴趣的 miRNA 的靶标模拟（TM）分子，将其整合到植物病毒载体中，并通过农杆菌浸润在马铃薯中表达，以直接结合感兴趣的内源性 miRNA 并阻断其功能。