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A rapid and efficient method for uniform gene expression using the barley stripe mosaic virus
1Département des sciences biologiques, Centre TOXEN, Université du Québec à Montréal, Montreal, QC H3C 3P8 Canada.
Plant Methods
|April 13, 2017
Summary
A new barley stripe mosaic virus (BSMV) seed imbibition method enables rapid and uniform gene expression in wheat and other plants. This simple technique allows for efficient gene function studies in the first generation of infected plants.
Area of Science:
- Plant molecular biology
- Virology
- Genetics
Background:
- Barley stripe mosaic virus (BSMV) is a common vector for studying gene function in cereals.
- Previous BSMV studies were limited to gene silencing in leaves, showing high variability.
- Overcoming these limitations required modifying the BSMV inoculation protocol for gene overexpression.
Purpose of the Study:
- To explore modifying the BSMV inoculation protocol for gene overexpression.
- To assess the efficiency of a novel seed imbibition method for BSMV delivery.
- To use an improved light, oxygen or voltage-sensing (iLOV) fluorescent protein as a reporter for BSMV infection and spread.
Main Methods:
- Tobacco (Nicotiana benthamiana) leaves were infected with BSMV via agroinfiltration.
- BSMV particles were extracted and used to inoculate wheat tissues.
- Two inoculation methods were compared: traditional leaf abrasion and seed imbibition.
Main Results:
- Seed imbibition resulted in high and uniform iLOV detection in wheat roots and leaves within 7 days.
- The method showed uniform expression across different wheat cultivars and other monocot/dicot species.
- Viral infection progression was stable in various organs and tissues and transmissible to the next generation.
Conclusions:
- BSMV effectively expresses small genes like iLOV in wheat roots and leaves.
- Seed imbibition provides rapid, uniform gene expression in multiple plant species, surpassing leaf abrasion.
- This method facilitates early gene function studies in the first plant generation, offering a simple, rapid, and cost-effective alternative to transgenic approaches.
