Protoplast Isolation and Transfection in Wheat.

Area of Science:

• Plant Science
• Molecular Biology
• Agricultural Biotechnology

Background:

• Wheat is a globally significant staple crop.
• Stable gene transfection in wheat remains challenging, necessitating alternative methods.
• Transient expression systems are vital for gene functional studies in wheat.

Purpose of the Study:

• To establish reliable protocols for wheat protoplast isolation and transfection.
• To demonstrate the utility of these protocols for gene functional analysis.
• To evaluate the efficiency of novel base editing tools in wheat protoplasts.

Main Methods:

• Protoplast isolation using cellulase R-10 and macerozyme R-10 enzymatic solutions.
• Polyethylene glycol (PEG)-mediated transfection for gene delivery into wheat protoplasts.
• Efficiency assessment of base editors using the developed protoplast system.

Main Results:

• Successfully isolated and transfected wheat protoplasts using the described enzymatic and PEG-mediated methods.
• Demonstrated the effectiveness of the protocol for evaluating gene function.
• Provided an example of assessing base editor efficiency in wheat protoplasts.

Conclusions:

• The developed protocols offer a versatile and accessible method for transient gene expression in wheat.
• These protocols facilitate both conventional gene function studies and the evaluation of genome editing reagents.
• This system is valuable for advancing genetic research and crop improvement in wheat.