Multiparameter in vivo imaging in plants using genetically encoded fluorescent indicator multiplexing.

Area of Science:

• Plant Biology
• Molecular Biology
• Biochemistry

Background:

• Biological processes in plants occur across diverse spatiotemporal scales.
• Understanding plant physiology requires analyzing processes from organismic to molecular levels in intact, living tissues.
• Fluorescent protein (FP)-based in vivo biosensing using genetically encoded fluorescent indicators (GEFIs) is a key technology.

Purpose of the Study:

• To provide an update on established direct FP-based biosensors in plants.
• To discuss the functional principles of GEFIs and their application in multiplexing.
• To highlight biological findings from GEFI-based multiparameter analyses and discuss challenges.

Main Methods:

• Utilizing genetically encoded fluorescent indicators (GEFIs) for in vivo biosensing.
• Employing GEFI multiplexing for simultaneous recording of multiple indicators.
• Conducting plate-reader-based analyses for multiparameter measurements in living plants.

Main Results:

• Established direct FP-based biosensors allow monitoring of ion, redox, sugar, hormone, ATP, and phosphatidic acid dynamics, and protein kinase activities.
• GEFI multiplexing enables in vivo multiparameter analyses by simultaneously recording various biological processes.
• Numerous biological findings have been achieved using GEFI-based multiplexing approaches.

Conclusions:

• GEFI-based multiplexing represents a powerful approach for in vivo multiparameter analyses in plants.
• This methodology advances the understanding of dynamic biological processes at cellular and molecular levels.
• Further development and application of GEFI biosensors will continue to yield significant biological insights.