Development of virus-induced genome editing methods in Solanaceous crops

Area of Science:

• Plant science
• Molecular biology
• Agricultural biotechnology

Background:

• CRISPR/Cas genome editing (GE) revolutionized plant mutagenesis but conventional methods require time-consuming transgenic lines.
• Virus-induced genome editing (VIGE) is effective in model plants but less explored in Solanaceous crops.

Purpose of the Study:

• To develop and optimize VIGE methods for Solanaceous plants (tomato, potato, eggplant).
• To establish both transgene-mediated and transgene-free VIGE approaches.
• To enhance VIGE efficiency using heat treatment.

Main Methods:

• Developed TRV-based VIGE in transgenic tomato expressing Cas9.
• Created PVX-based VIGE for transgene-free genome editing.
• Designed sgRNAs targeting Phytoene desaturase and optimized VIGE conditions.
• Applied heat treatment (37°C) to enhance editing efficiency.
• Used deep sequencing to evaluate mutation rates.
• Regenerated edited plants via tissue culture.

Main Results:

• Achieved 40.3% (TRV) and 36.5% (PVX) mutation rates in tomato targeting Phytoene desaturase.
• Heat treatment significantly increased editing efficiency: 33-46% (TRV) and 56-76% (PVX).
• Successfully applied PVX-mediated VIGE to potato and eggplant.

Conclusions:

• Developed efficient TRV- and PVX-based VIGE systems for Solanaceous plants.
• PVX-mediated VIGE offers a transgene-free genome editing approach.
• VIGE methods, enhanced by heat treatment, provide a rapid and efficient tool for crop improvement.