Both methylerythritol phosphate and mevalonate pathways contribute to biosynthesis of each of the major isoprenoid classes in young cotton seedlings

Abstract

In higher plants, both the methylerythritol phosphate (MEP) and mevalonate (MVA) pathways contribute to the biosynthesis of isoprenoids. However, despite a significant amount of research on the activity of these pathways under different conditions, the relative contribution of each to the biosynthesis of diverse isoprenoids remains unclear. In this work, we examined the formation of several classes of isoprenoids in cotton (Gossypium hirsutum L.). After feeding [5,5-(2)H2]-1-deoxy-D-xylulose ([5,5-(2)H2]DOX) and [2-(13)C]MVA to intact cotton seedlings hydroponically, incorporation into isoprenoids was analyzed by MS and NMR. The predominant pattern of incorporation followed the classical scheme in which C5 units from the MEP pathway were used to form monoterpenes (C10), phytol side chains (C20) and carotenoids (C40) while C5 units from the MVA pathway were used to form sesquiterpenes (C15), terpenoid aldehydes (C15 and C25) and steroids/triterpenoids (C30). However, both pathways contributed to all classes of terpenoids, sometimes substantially. For example, the MEP pathway provided up to 20% of the substrate for sterols and the MVA pathway provided as much as 50% of the substrate for phytol side chains and carotenoids. Incorporation of C5 units from the MEP pathway was highest in cotyledons, compared to true leaves, and not observed at all in the roots. Incorporation of C5 units from the MVA pathway was highest in the roots (into sterols) and more prominent in the first true leaves than in other above-ground organs. The relative accumulation of label in intermediates vs. end products of phytosterol metabolism confirmed previous identification of slow steps in this pathway.

Keywords: 1-deoxy-d-xylulose; Carotenoids; Cross-talk; DMAPP; DOX; DXS; Gossypium hirsutum L. (cotton); HMGR; IPP; Isoprenoids; MEP; MEP pathway; MVA; MVA pathway; Plant biochemistry; Sterols; Terpenoids; deoxyxylulose-5-phosphate synthase; dimethylallyl diphosphate; hydroxymethylglutaryl coenzyme A reductase; isopentenyl diphosphate; methylerythritol phosphate; mevalonate.