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Related Experiment Video

Updated: Nov 1, 2025

Quantification of Endogenous Auxin and Cytokinin During Internode Culture of Ipecac
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Protocol: analytical methods for visualizing the indolic precursor network leading to auxin biosynthesis.

Molly Tillmann1, Qian Tang2, Jerry D Cohen2

  • 1Department of Horticultural Science and Microbial and Plant Genomics Institute, University of Minnesota, St. Paul, MN, USA. mkreiser@umn.edu.

Plant Methods
|June 23, 2021
PubMed
Summary

Researchers developed new methods to track how plants make auxin (indole-3-acetic acid), a key plant hormone. This helps understand auxin biosynthesis pathways under different conditions.

Keywords:
Auxin biosynthesisLC–MSMetabolic inhibitorsPathway analysisStable isotope labeling

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Area of Science:

  • Plant Biology
  • Biochemistry

Background:

  • Auxin (indole-3-acetic acid) is a vital plant hormone regulating growth and environmental responses.
  • Several indole-3-acetic acid (IAA) biosynthesis pathways exist, but their in vivo utilization is poorly understood.

Purpose of the Study:

  • To develop and validate techniques for analyzing auxin biosynthesis pathways in plants.
  • To investigate the in vivo utilization of different IAA biosynthetic routes.

Main Methods:

  • Utilized stable isotope-labeled precursors to trace metabolic flux.
  • Employed chemical inhibitors to selectively block auxin synthesis steps.
  • Applied liquid chromatography-mass spectrometry (LC-MS) for sensitive detection and quantification.

Main Results:

  • Demonstrated the efficacy of stable isotope tracing combined with chemical inhibition.
  • Established a robust method for analyzing auxin biosynthesis in Arabidopsis thaliana seedlings.
  • Provided a framework for characterizing novel auxin biosynthetic routes.

Conclusions:

  • The described techniques enable detailed in vivo analysis of IAA biosynthesis.
  • These methods are applicable to various plant tissues and experimental conditions.
  • Facilitates a deeper understanding of plant hormone regulation and adaptation.