Barley Stripe Mosaic Virus-Mediated Tools for Investigating Gene Function in Cereal Plants and Their Pathogens: Virus-Induced Gene Silencing, Host-Mediated Gene Silencing, and Virus-Mediated Overexpression of Heterologous Protein

A New Barley Stripe Mosaic Virus Allows Large Protein Overexpression for Rapid Function Analysis

“…Recently, we 179 demonstrated that a seed imbibition method leads to a more uniform and rapid expression of the 180 smaller iLOV marker compared to the leaf abrasion inoculation method (Cheuk and Houde,181 2017). The uneven GFP signal distribution reported in previous studies (Lawrence and Jackson,182 2001; Lee et al, 2012) could be due to the inoculation method and/or instability caused by the 183 insert size within the RNAγ genome. To verify the cargo limit of the three-component BSMV 184 system, the iLOV (336 bp) and GFP cDNAs (720 bp) were inserted into the γ genome vector or 185 in the γ2 genome vector (Fig.…”

“…The three-component BSMV system has a low cargo capacity 175 Transformation using the three-component BSMV system and leaf abrasion inoculation method 176 to express GFP was shown to yield patchy fluorescence throughout the infected leaf tissue 177 (Lawrence and Jackson, 2001;Lee et al, 2012), suggesting instability of large inserts within the 178 γ genomic BSMV RNA Lee et al, 2012). Recently, we 179 demonstrated that a seed imbibition method leads to a more uniform and rapid expression of the 180 smaller iLOV marker compared to the leaf abrasion inoculation method (Cheuk and Houde,181 2017).…”

Secreted biomolecules in fungal plant pathogenesis

“…However, the availability of fully sequenced pathogen genomes and next-generation sequencing transcriptome analysis of infection time courses is leading to the identification of the in planta expressed small gene clusters potentially responsible for secondary metabolite production. The application of virus-induced gene silencing (VIGS) to assess the plant host contribution to the interaction and also identify the specific interaction targets of each secreted biomolecule is now available for many plant species, representing a significant technological advancement (Lee et al, 2012(Lee et al, , 2014. These types of plants studies that often also use transformed fungal strains with an altered effector complement will truly enable the dissection of the communication between pathogen and host.…”

The impacts of BSMV on vegetative growth and water status in hulless barley (Hordeum vulgare var. nudum) in VIGS study

“…Previous studies showed that VIGS persists for longer if there is a continuous presence of the virus [5,32,33], however if the viral infection symptoms disappeared, the subsequent gene silencing phenotype would also disappear. It was reported that the most efficient time period for VIGS is within 3 weeks [34] and the efficiency of VIGS decreased after 1 month when plants start to recover from silencing [22].…”

“…This method has several advantages over conventional transgenic technologies in that it is easy to manipulate, saves time and has a low cost [4]. Barley stripe mosaic virus (BSMV) is a Hordeivirus which can cause mild to moderate mosaic symptoms [4,5]. It has been genetically reconstructed to a VIGS vector for monocots [6] and has been widely utilized in gene silencing [6][7][8][9][10][11][12][13].…”