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Optimized LC-MS method for simultaneous polyamine profiling and ADC/ODC activity quantification and evidence that
Erin Samantha Ritchie1, Edda von Roepenack-Lahaye2, Dennis Perrett1
1ZMBP - General Genetics, University of Tübingen, Tuebingen, Germany.
Frontiers in Plant Science
|September 29, 2025
Summary
A new liquid chromatography-mass spectrometry method accurately measures plant polyamine enzyme activity. This tool helps understand polyamine homeostasis and its critical role in plant development, like flower formation.
Area of Science:
- Plant Biology
- Biochemistry
- Metabolomics
Background:
- Polyamines (PAs) are crucial for plant growth and stress response, but their regulation is complex.
- Transcript-based methods fail to predict polyamine-synthesizing enzyme levels accurately due to post-transcriptional regulation.
Purpose of the Study:
- To develop and optimize a sensitive LC-MS method for quantifying key polyamine-synthesizing enzyme activities (ADC and ODC).
- To establish a toolkit for studying polyamine homeostasis and its impact on plant development.
Main Methods:
- Optimized a liquid chromatography-mass spectrometry (LC-MS) assay using stable isotope substrates to measure arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) activity.
- Adapted the method for heterologous expression in *Nicotiana benthamiana* and analyzed tomato CRISPR mutants.
Main Results:
- Achieved >10-fold improvement in assay sensitivity in tomato leaf tissue.
- Demonstrated a correlation between ADC protein abundance and activity in *N. benthamiana*, enabling inference of native protein levels.
- Quantified 11 PA-network metabolites, providing a comprehensive metabolic overview.
- Identified severe developmental defects, including flower loss, in tomato *adc1/adc2* mutants.
Conclusions:
- The optimized LC-MS method offers a robust approach to study polyamine homeostasis.
- Functional analysis of recombinant enzymes and CRISPR mutants provides insights into PA-dependent developmental processes, particularly flower development.
