Introduction of the Ribo-BiFC method to plants using a split mVenus approach

Area of Science:

• Plant Molecular Biology
• Biochemistry
• Cell Biology

Background:

• Translation is a vital cellular process, crucial for plant development and environmental responses.
• Measuring translation rates in living plant tissues in vivo remains technically challenging.

Purpose of the Study:

• To introduce and validate a novel in vivo translation marker for plant tissues.
• To enable visualization of translation dynamics in real-time within plants.

Main Methods:

• Developed the Ribo-BiFC system by combining fruit fly-derived methods with a plant-optimized split-mVenus bimolecular fluorescence complementation (BiFC) system.
• Tagged Arabidopsis thaliana small and large subunit ribosomal proteins (RPS and RPL) with mVenus fluorescent protein fragments.
• Tested Ribo-BiFC in Nicotiana benthamiana epidermal cells using transient expression.

Main Results:

• Successfully visualized complemented mVenus fluorescence, indicating functional BiFC tagging of ribosomal proteins.
• Demonstrated the potential of Ribo-BiFC to visualize the proximity of assembled 80S ribosomal subunits during translation.
• Observed a detectable, albeit less intense, signal compared to known interactors, confirming system functionality.

Conclusions:

• The Ribo-BiFC system offers a promising tool for visualizing translational rates in plant tissues.
• This approach can be utilized in stable transgenic lines to study dynamic changes in translation during plant development and stress.
• Ribo-BiFC has potential applications in diverse genetic backgrounds and under various abiotic stress conditions.