Andrographolide, a diterpene lactone, is isolated from Andrographis paniculata which is well known for its medicinal properties. The biosynthetic route to andrographolide was studied using [1-(13)C]acetate, [2-(13)C]acetate and [1,6-(13)C(2)]glucose. The peak enrichment of eight carbon atoms in the (13)C NMR spectra of andrographolide suggested that deoxyxylulose pathway (DXP) is the major biosynthetic pathway to this diterpene. The contribution of the mevalonic acid pathway (MVA) is indicated by the observed (13)C-labeling pattern, and because the labeling patterns indicate a simultaneous contribution of both methyl erythritol phosphate (MEP) and MVA pathways it can be deduced that cross-talk occurs between plastids and cytoplasm.