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.
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