Fig. 5.

Silencing of AbCYP82M3 disrupts tropane and pyrrolidine alkaloid biosynthesis. a The tropane and pyrrolidine alkaloid biosynthetic pathway in the Solanaceae starting from putrescine (1). Experimentally validated enzymes are shown in blue, AbPYKS and AbCYP82M3 are shown in red, and unknown reactions are noted with question marks. Enzyme abbreviations are as follows: PMT putrescine methyltransferase, MPO methylputrescine oxidase, TRI tropinone reductase I, TRII tropinone reductase II, CYP80F1 littorine mutase, H6H hyoscyamine-6-hydroxylase. Formation of cuscohygrine (9) via nonenzymatic decarboxylative condensation as per Supplementary Figure 5 is shown in red. b Abundance of tropane and pyrrolidine alkaloids together with selected precursors in TRV2 empty vector control lines (cyan bars) and AbCYP82M3 VIGS lines (red bars). Data are presented as the mean n = 21 biological replicates. Asterisks denote significant differences (***p < 0.001) as determined by Student’s t test. For each box plot, the lower and upper bounds of the box indicate the first and third quartiles, the line indicates the median value, and the whiskers extend to the minimum and maximum data points. This experiment was replicated twice. Source Data are provided as a Source Data file