A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell...

Area of Science:

• Plant Biology
• Molecular Biology
• Cell Biology

Background:

• Plant protoplasts are crucial for gene characterization and signaling pathway analysis.
• Programmed cell death (PCD) is a vital process in plant development.
• Maize nucellus undergoes PCD during embryo sac development, but nucellus protoplasts have not been previously utilized.

Purpose of the Study:

• To establish conditions for isolating and transfecting maize nucellus protoplasts.
• To demonstrate the utility of this system for protein expression, localization, and interaction studies.
• To investigate programmed cell death (PCD)-related processes in maize.

Main Methods:

• Isolation and transfection of maize nucellus protoplasts.
• Transient gene expression assays (e.g., MADS29).
• Quantitative real-time PCR, β-glucuronidase, and luciferase activity assays.

Main Results:

• Successfully established protocols for maize nucellus protoplast isolation and transfection.
• Demonstrated the system's effectiveness for protein expression, localization, and interaction.
• Transient MADS29 expression increased Cys-protease gene transcripts and enhanced promoter activity.

Conclusions:

• A highly efficient maize nucellus protoplast system for transient gene expression was developed.
• This system is suitable for investigating PCD-related processes in maize.
• The findings open new avenues for studying plant development and gene function.