Stable Isotope Labeling and Quantification of Photosynthetic Metabolites

Methods Mol Biol. 2024:2790:439-466. doi: 10.1007/978-1-0716-3790-6_24.

Abstract

Stable isotope labeling with 13CO2 coupled with mass spectrometry allows monitoring the incorporation of 13C into photosynthetic intermediates and is a powerful technique for the investigation of the metabolic dynamics of photosynthesis. We describe here a protocol for 13CO2 labeling of large leaved plants and of Arabidopsis thaliana rosette, and a method for quantitative mass spectrometry analyses to uncover the labeling pattern of Calvin-Benson cycle sucrose, and starch synthesis as well as carbon-concentrating mechanism metabolites.

Keywords: 13C enrichment; 13CO2 labeling; Calvin-Benson Cycle; Carbon-concentrating mechanism; Isotopomer distribution; LC-MS/MS; Mass spectrometry; Metabolite quantification; Photosynthesis; Pulse-chase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis* / metabolism
  • Carbon Dioxide / metabolism
  • Carbon Isotopes* / metabolism
  • Isotope Labeling* / methods
  • Mass Spectrometry / methods
  • Metabolomics / methods
  • Photosynthesis*
  • Plant Leaves / metabolism
  • Starch / metabolism
  • Sucrose / metabolism

Substances

  • Carbon Isotopes
  • Sucrose
  • Carbon Dioxide
  • Starch