U-13C-glucose incorporation into source leaves of Brassica napus highlights light-dependent regulations of metabolic fluxes within central carbon metabolism

• Published in: Journal of plant physiology 2024-01, Vol.292, p.154162-154162, Article 154162
• Main Authors: Dellero, Younès, Berardocco, Solenne, Bouchereau, Alain
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Brassica napus; Carbon isotopologue distribution; Chemical Sciences; Dark; Glycolysis; Isotope; Life Sciences; Light; Serine; Tricarboxylic acid cycle; Vegetal Biology
• ISSN: 0176-1617; 1618-1328
• DOI: 10.1016/j.jplph.2023.154162

Plant central carbon metabolism comprises several important metabolic pathways acting together to support plant growth and yield establishment. Despite the emergence of 13C-based dynamic approaches, the regulation of metabolic fluxes between light and dark conditions has not yet received sufficient attention for agronomically relevant plants. Here, we investigated the impact of light/dark conditions on carbon allocation processes within central carbon metabolism of Brassica napus after U-13C-glucose incorporation into leaf discs. Leaf gas-exchanges and metabolite contents were weakly impacted by the leaf disc method and the incorporation of glucose. 13C-analysis by GC-MS showed that U-13C-glucose was converted to fructose for de novo biosynthesis of sucrose at similar rates in both light and dark conditions. However, light conditions led to a reduced commitment of glycolytic carbons towards respiratory substrates (pyruvate, alanine, malate) and TCA cycle intermediates compared to dark conditions. Analysis of 13C-enrichment at the isotopologue level and metabolic pathway isotopic tracing reconstructions identified the contribution of multiple pathways to serine biosynthesis in light and dark conditions. However, the direct contribution of the glucose-6-phosphate shunt to serine biosynthesis was not observed. Our results also provided isotopic evidences for an active metabolic connection between the TCA cycle, glycolysis and photorespiration in light conditions through a rapid reallocation of TCA cycle decarboxylations back to the TCA cycle through photorespiration and glycolysis. Altogether, these results suggest the active coordination of core metabolic pathways across multiple compartments to reorganize C-flux modes. •Leaf discs show photosynthetic and respiratory activities similar to those of attached leaves.•The availability of glycolytic by-products is a major regulator of tricarboxylic acid (TCA) cycle activity in the light.•TCA cycle decarboxylations can be reintroduced into this cycle through photosynthetic reassimilation followed by glycolysis.