A Plant Virus-Based Vector System for Gene Function Studies in Pepper1

• Published in: Plant physiology (Bethesda) Vol. 181; no. 3; pp. 867 - 880
• Main Authors: Choi, Boram, Kwon, Sun-Jung, Kim, Myung-Hwi, Choe, Siwon, Kwak, Hae-Ryun, Kim, Mi-Kyeong, Jung, Choonkyun, Seo, Jang-Kyun
• Format: Journal Article
• Language: English
• Published: American Society of Plant Biologists 03.09.2019
• Subjects: Breakthrough Technologies
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.19.00836

A plant virus-based system enables transient gain- and loss-of-function studies in pepper, a highly recalcitrant species for genetic transformation. While pepper ( Capsicum annuum ) is a highly recalcitrant species for genetic transformation studies, plant virus-based vectors can provide alternative and powerful tools for transient regulation and functional analysis of genes of interest in pepper. In this study, we established an effective virus-based vector system applicable for transient gain- and loss-of-function studies in pepper using Broad bean wilt virus2 (BBWV2). We engineered BBWV2 as a dual gene expression vector for simultaneous expression of two recombinant proteins in pepper cells. In addition, we established enhanced and stable expression of recombinant proteins from the BBWV2-based dual vector via coexpression of a heterologous viral suppressor of RNA silencing. We also developed a BBWV2-based virus-induced gene silencing (VIGS) vector, and we successfully silenced the phytoene desaturase gene ( PDS ) using the BBWV2-based VIGS vector in various pepper cultivars. Additionally, we optimized the BBWV2-based VIGS system in pepper by testing the efficiency of PDS gene silencing under different conditions. This BBWV2-based vector system represents a convenient approach for rapid and simple analysis of gene functions in pepper.