02568nam a2200265Ia 4500 MX-MdCICY 20260521091801.0 CICY B-21371 Using targeted metabolomics to elucidate the indole auxin network in plants Methods in Enzymology. 676, 239-278, 2022, DOI: 10.1016/bs.mie.2022.07.038 The plant hormone auxin plays important roles throughout the entire life span of a plant and facilitates its adaptation to a changing environment. Multiple metabolic pathways intersect to control the levels and flux through indole-3-acetic acid (IAA), the primary auxin in most plant species. Measurement of changes in these pathways represents an important objective to understanding core aspects of auxin signal regulation. Such studies have become approachable through the technologies encompassed by targeted metabolomics. By monitoring incorporation of stable isotopes from labeled precursors into proposed intermediates, it is possible to trace pathway utilization and characterize new biosynthetic routes to auxin. Chemical inhibitors that target specific steps or entire pathways related to auxin synthesis aid these techniques. Here we describe methods for obtaining stable isotope labeled pathway intermediates necessary for pathway analysis and quantification of compounds. We describe how to use isotope dilution with methods employing either gas chromatography or high performance liquid chromatography mass spectrometry techniques for sensitive analysis of IAA. Complete biosynthetic pathway analysis in seedlings using multiple stable isotope-labeled precursors and chemical inhibitors coupled with highly sensitive liquid chromatography-mass spectrometry methods are described that allow rapid measurement of isotopic flux into biochemical pools. These methods should prove to be useful to researchers studying aspects of the auxin metabolic network in vivo in a variety of plant tissues and during various environmental conditions. © 2022 Elsevier Inc. AUXIN BIOSYNTHESIS ISOTOPE DILUTION ANALYSIS MASS SPECTROMETRY METABOLIC INHIBITORS PATHWAY ANALYSIS STABLE ISOTOPE LABELING Cohen J.D. Tang Q. Hegeman A.D. https://drive.google.com/file/d/19zRDxW-qRv2NZGcXdX7WJ8DVLI8VkLWt/view?usp=drivesdk Para ver el documento ingresa a Google con tu cuenta @cicy.edu.mx Loc REF1 250602s9999 xx |||||s2 |||| ||und|d 31371 31371 0 0 Loc 0 0 F1 CICY CICY RE 2025-06-25 0 B-21371 2025-06-25 16:43:53 2025-06-25 REF1