Molecular Plant Biology and Biotechnology Lab, CSIR-Central Institute of Medicinal and Aromatic Plants, Research Centre, Bengaluru, Karnataka, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India.
Methods Mol Biol. 2022;2408:147-163. doi: 10.1007/978-1-0716-1875-2_10.
Virus-induced gene silencing (VIGS) is a functional genomics tool to transiently downregulate the expression of target gene(s) by exploiting the plant's innate defense mechanism against invading RNA viruses. VIGS is a rapid and efficient approach to analyze the gene function, particularly, in the plants that are not amenable to stable genetic transformation. This strategy has been successfully used to decipher the function of several genes and transcription factors involved in the biosynthesis of specialized metabolites and regulation of specialized metabolism, respectively, in different medicinal and aromatic plants. Here, we describe a detailed Tobacco rattle virus (TRV)-mediated VIGS protocol for silencing of the gene encoding Phytoene desaturase (PDS) in important medicinal plants Catharanthus roseus, Calotropis gigantean, Rauwolfia serpentina, and Ocimum basilicum. Our methods allow the study of gene function within 3-4 weeks after agro-inoculation, and can be an easy and efficient approach for future studies on understanding of the biosynthesis of specialized metabolites in these important medicinal plants.
病毒诱导的基因沉默（VIGS）是一种功能基因组学工具，通过利用植物对入侵 RNA 病毒的先天防御机制，暂时下调靶基因（s）的表达。VIGS 是一种快速有效的方法来分析基因功能，特别是在那些不易进行稳定遗传转化的植物中。该策略已成功用于破译参与次生代谢物生物合成和次生代谢调控的几个基因和转录因子的功能，分别在不同的药用和芳香植物中。在这里，我们描述了一个详细的烟草脆裂病毒（TRV）介导的 VIGS 协议，用于沉默重要药用植物长春花、大花犀角、萝芙木和罗勒中编码类胡萝卜素脱饱和酶（PDS）的基因。我们的方法允许在农杆菌接种后 3-4 周内研究基因功能，并且可以成为未来研究这些重要药用植物中次生代谢物生物合成的一种简单有效的方法。
