Metabolic flux analysis: recent advances in carbon metabolism in plants

Isotopic tracers are used to both trace metabolic pathways and quantify fluxes through these pathways. The use of different labeling methods recently led to profound changes in our views of plant metabolism. Examples are taken from primary metabolism, with sugar interconversions, carbon partitioning between glycolysis and the pentose phosphate pathway, or metabolite inputs into the tricarboxylic acid (TCA) cycle, as well as from secondary metabolism with the relative contribution of the plastidial and cytosolic pathways to the biosynthesis of terpenoids. While labeling methods are often distinguished according to the instruments used for label detection, emphasis is put here on labeling duration. Short time labeling is adequate to study limited areas of the metabolic network. Long-term labeling, when designed to obtain metabolic and isotopic steady-state, allows to calculate various fluxes in large areas ofcentral metabolism. After longer labeling periods, large amounts of label accumulate in structural or storage compounds: their detailed study through the retrobiosynthetic method gives access to the biosynthetic pathways of otherwise undetectable precursors. This chapter presents the power and limits of the different methods, and illustrates how they can be associated with each other and with other methods of cell biology, to provide the information needed for a rational approach of metabolic engineering.