Enzyme Inhibitor Studies Reveal Complex Control of Methyl-D-Erythritol 4-Phosphate (MEP) Pathway Enzyme Expression in Catharanthus roseus
Figure 3
Effect of paraquat treatment on the expression of MEP pathway genes at protein and transcript level, respectively, in leaf discs obtained from mature leaves of 6-week-old C. roseus plants.
Leaf discs (5 mm diameter) were collected from fully expanded leaves and floated on distilled water in the presence or absence of 0.5 µM paraquat for the indicated time intervals at 25°C under continuous light (270 µmol m−2 s−1). (A) DXS, DXR and HDS proteins were detected by immunoblot. Equal sample loading was confirmed by Coomassie staining. The arrow marks the position of mature CrDXS2A protein.(B) transcript amounts for DXS (isoforms 1, 2A & 2B), DXR and HDS were determined by qPCR relative to the geometric mean of multiple reference genes according to Vandesompele et al. [60]. The experiment was performed 3 times; values from a representative experiment are presented ± SD.