Phenotype microarray-based assessment of metabolic variability in plant protoplasts

Area of Science:

• Plant Science
• Metabolomics
• Cell Biology

Background:

• Plant productivity is shaped by genetics and environment, necessitating advanced phenotyping methods.
• Cellular-level phenotyping is crucial for understanding metabolic bases of plant phenotypes and genetic modifications.
• Phenotype Microarray (PM) is a high-throughput cellular metabolic characterization tool, but not yet established for plant cells.

Purpose of the Study:

• To develop and validate a Phenotype Microarray (PM) procedure for plant cell metabolic characterization.
• To establish a high-throughput system for analyzing cellular metabolism in plant protoplasts.
• To enable comparative metabolic studies at the cellular level across different plant varieties and genetic backgrounds.

Main Methods:

• Adapted Biolog's Phenotype Microarray (PM) technology for plant cell analysis using isolated protoplasts.
• Optimized conditions for protoplast isolation, purification, and inoculation into PM microplates.
• Utilized redox potential markers to monitor cellular respiration and metabolic activity in response to various substrates and stresses (e.g., NaCl).

Main Results:

• Successfully demonstrated the feasibility of using PM technology for metabolic profiling of plant protoplasts.
• Identified optimal conditions for PM testing on protoplasts from *Solanum tuberosum* L. and *S. lycopersicum* L.
• Evaluated plant protoplast responses to different NaCl concentrations and toxic compounds using the developed PM system.

Conclusions:

• A standardized, high-throughput system for plant protoplast metabolic characterization has been successfully developed.
• This method provides a foundation for plant cell metabolic phenotyping, facilitating comparative studies.
• The approach supports research on cultivars, species, wild-type organisms, and genome-edited plants at the cellular level.