TRV-GFP: a modified Tobacco rattle virus vector for efficient and visualizable analysis of gene function

Area of Science:

• Plant molecular biology
• Gene function analysis
• Virus-induced gene silencing (VIGS)

Background:

• Virus-induced gene silencing (VIGS) is crucial for plant gene characterization.
• Traditional Tobacco rattle virus (TRV) vectors show low efficiency in non-Solanaceae plants, like economically important roses.
• A traceable VIGS tool is needed for these species.

Purpose of the Study:

• To develop an easily traceable TRV vector for enhanced VIGS efficiency.
• To assess the silencing efficacy of the modified TRV-GFP vector in various plant species.
• To establish GFP as an indicator for successful gene silencing.

Main Methods:

• A green fluorescent protein (GFP) gene was integrated into the TRV2 vector (TRV-GFP).
• TRV-GFP infection efficiency was tested in Nicotiana benthamiana, Arabidopsis thaliana, rose, strawberry, and chrysanthemum.
• TRV-GFP was used to silence the phytoene desaturase (PDS) gene in roses, and GFP expression was monitored.

Main Results:

• TRV-GFP exhibited infection efficiency comparable to the original TRV vector across tested plants.
• GFP expression allowed easy monitoring of TRV spread using fluorescence microscopy and UV lamps.
• Silencing of the PDS gene in roses resulted in a photobleached phenotype in 75-80% of GFP-positive plants.
• GFP abundance correlated negatively with PDS gene levels, indicating its utility as a silencing indicator.

Conclusions:

• The TRV-GFP vector provides a visualizable and efficient tool for VIGS.
• This method is valuable for predicting successful gene silencing in plants.
• The TRV-GFP system is particularly beneficial for functional gene studies in non-Solanaceae plants.