Genetically Encoded Fluorescent Biosensors Enable Noninvasive Real-Time Visualization of Nitrate Dynamics in Intact...

Area of Science:

• Plant Biology
• Biochemistry
• Molecular Biology

Background:

• Nitrate (NO3-) is crucial for plant growth, acting as both a nutrient and signaling molecule.
• Understanding nitrate's spatiotemporal dynamics is key to plant nitrogen metabolism research.
• Current detection methods lack real-time, non-invasive, whole-plant monitoring capabilities.

Purpose of the Study:

• To develop a novel, genetically encoded biosensor for real-time nitrate detection in plants.
• To enable non-invasive, spatiotemporal imaging of nitrate dynamics in living plants.
• To advance the study of plant nitrogen metabolism and inform crop improvement.

Main Methods:

• Constructed a nitrate biosensor (NitNRCL1) using a split firefly luciferase complementation system.
• Validated NitNRCL1 functionality in prokaryotic and eukaryotic systems, including diverse plant species.
• Utilized NitNRCL1 for non-invasive, real-time imaging of nitrate signaling in *Arabidopsis thaliana*.

Main Results:

• NitNRCL1 successfully responds to varying nitrate availability.
• The biosensor generates stable chemiluminescent signals in bacteria and plants.
• Demonstrated real-time, whole-plant imaging of nitrate spatiotemporal dynamics in *Arabidopsis thaliana*.

Conclusions:

• NitNRCL1 is a robust, novel tool for investigating plant nitrate transport, signaling, and metabolism.
• The biosensor enhances mechanistic understanding of plant nitrate biology.
• Provides a foundation for developing nitrogen-use-efficient crops and precision fertilization strategies.