Extended Data Fig. 7: Expression of the C. roseus strictosidine exporter gene CrNPF2.9 boosts C. ipecacuanha protoemetine biosynthesis and enables biosynthesis from dopamine and secologanin directly.

a, comparison of protoemetinol accumulation upon expression of indicated C. ipecacuanha biosynthesis genes in N. benthamiana with or without co-expression of transporter CrNPF2.9. Exogenous DAI/DAII are infiltrated as substrate and allowed to react for 24 hours before the leaf was harvested. Data was extracted from experiment shown in Supplementary Fig. 10. Peak areas are shown here relative to areas from samples where CrNPF2.9 was not expressed as bars of the mean of three replicates. b, expression of CiIps with or without co-overexpression of CrNPF2.9 did not lead to enhanced accumulation of the R-epimer derivative ipecoside. c, when C. ipecacuanha protoemetine pathway genes are expressed and uncoupled secologanin and dopamine are fed for 48 hours, only traces of protoemetinol can be detected (indicated with an asterisk). When CrNPF2.9 is co-overexpressed protoemetinol is formed to detectable levels. d, it is hypothesized that spontaneous in planta coupling occurs in the vacuole and the resulting formed DAII is only accessible to the cytosolic pathway enzymes if it is exported by a vacuolar exporter. CrNPF2.9 appears to export DAII. These results suggest the presence of a yet to be identified vacuolar exporter in C. ipecacuanha and A. salviifolium. LC-MS peak areas are shown as bars of the mean of three biological replicates, error bars are standard error of the mean.