Fig. 7: In planta reconstitution of nicotine biosynthesis.
From: Nicotine biosynthesis is completed by cryptic activating glucosylation
Leaves of N. benthamiana were agroinfiltrated with a combination of nicotine biosynthesis genes (day 0), fed with nicotinamide-d4 (ND-d4) at day 4, and harvested for metabolite analysis at day 6. The gene combination producing the N-methylpyrrolinium (ODC, PMT and MPO) is abbreviated as NMP. A Accumulation of the labelled compounds is indicated at the top (d3 and d4 versions together). Each row corresponds to the gene combination shown in the matrix on the left (green/white indicates presence/absence). Traces are representative extracted ion chromatograms (EICs, m/z ± 0.005) corresponding to the labelled compounds (black) in comparison to chemically synthesized or commercial standards (blue) (see Supplementary Fig. 12 for MS2 spectra). Signal intensities are normalized to the largest peak on each column; standards were rescaled to match the largest peak. The nicotine and anabasine standards were analysed as a mixed sample. The insert on the right shows the chiral analysis of nicotine in the presence of all genes and in the NicGS drop-out (D.O.). B Individual quantification of d3 and d4 isotopologues corresponding to the compounds and gene combinations shown in (A). Bars show mean EIC peak area (n = 8); points are individual replicates; error bars show standard deviation (see statistics in Supplementary Data 11). Source data are provided as a Source Data file.
