Vacuum and Co-cultivation Agroinfiltration of (Germinated) Seeds Results in Tobacco Rattle Virus (TRV) Mediated Whole-Plant Virus-Induced Gene Silencing (VIGS) in Wheat and Maize

Abstract

Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) has been frequently used in dicots. Here we show that it can also be used in monocots, by presenting a system involving use of a novel infiltration solution (containing acetosyringone, cysteine, and Tween 20) that enables whole-plant level VIGS of (germinated) seeds in wheat and maize. Using the established system, phytoene desaturase (PDS) genes were successfully silenced, resulting in typical photo-bleaching symptoms in the leaves of treated wheat and maize. In addition, three wheat homoeoalleles of MLO, a key gene repressing defense responses to powdery mildew in wheat, were simultaneously silenced in susceptible wheat with this system, resulting in it becoming resistant to powdery mildew. The system has the advantages generally associated with TRV-mediated VIGS systems (e.g., high-efficiency, mild virus infection symptoms, and effectiveness in different organs). However, it also has the following further advantages: (germinated) seed-stage agroinfiltration; greater rapidity and convenience; whole-plant level gene silencing; adequately stable transformation; and suitability for studying functions of genes involved in seed germination and early plant development stages.

Host-Induced Gene Silencing of an Adenylate Kinase Gene Involved in Fungal Energy Metabolism Improves Plant Resistance to Verticillium dahliae

“…Recently, HIGS has also proved to be a promising strategy for silencing a foreign gene in a host to investigate numerous plant-pathogen systems [61,62], including Vd in various hosts [33,34,63]. In the present study, positive plants at 10 days after infiltration showed distinct photobleaching, suggesting the massive presence of dsRNA and its active role in gene silencing [64][65][66]. In our study, seedlings that were inoculated with Vd developed varying degrees of wilting from 10 to 12 dpi.…”

Pseudomonas fluorescens MZ05 Enhances Resistance against Setosphaeria turcica by Mediating Benzoxazinoid Metabolism in the Maize Inbred Line Anke35

“…A virus-mediated gene silencing system in maize plants was developed as recently reported by Zhang et al [27]. The VIGS vectors, including pTRV1 and pTRV2, were used for this study.…”

A roadmap for gene functional characterisation in wheat

“…Transient gene silencing through Virus Induced Gene Silencing (VIGS) has also be performed in wheat primarily to investigate disease resistance. VIGS has been carried out in a range of varieties, but is generally restricted to a few tissue types such as leaf tissues (Lee et al, 2015), young seedlings (Zhang et al, 2017a) and spikes (Ma et al, 2012).…”