Fig. 4: The evolution of the tropine gene cluster is associated with tropine diversity across the Solanaceae family.
a The tropine gene cluster syntenic regions of seven Solanaceae species and the outgroup species Ipomea trifida (Convolvulaceae). Genes encoding TRI are indicated in the red rectangle, and genes encoding CYP82M3 are indicated in the green rectangle. b Analysis of tropine in plants by combined LC/MS. The roots, stems and leaves of plants were analyzed for the detection of tropine. The extracted ion chromatograms of tropine are highlighted in red. c Extracted ion chromatograms showing the in vitro activity of seven purified recombinant TRIs from A. belladonna (AbTRI), D. stramonium (DsTRI), C. annuum (CaTRI), S. lycopersicum (SlTRI), S. tubersoum (StTRI), SlTRIL169V, and StTRIL169V with tropinone used as substrate. d Extracted ion chromatograms showing the in vivo activity of CaCYP82M3 and SlCYP82M3 in tobacco leaves. e, f The evaluation of enzymatic kinetics on five wild-type TRIs and two mutants by Kcat / Km values (e) or by Km values (f). The raw Michaelis–Menten curves of these TRIs for the NADPH-dependent reduction reaction of tropinone are presented in Fig. S31. The data are presented as means values +/− s.d. (n = 3 biologically independent samples). Different letters in e and f indicate significantly different values compared with each other at P < 0.05 analyzed by two-sided Student’s t-test. ND not detectable. Source data underlying e and f are provided as a Source Data file.
