Fig. 5: Fruit of game36 mutant tomato lines display significantly altered SGA metabolism.

a–d Levels of SGAs produced upstream of the GAME36 reaction step (a, c), and downstream of the GAME36 reaction step (b, d) in green (a, b) and red (c, d) fruit of game36 mutant tomato lines as compared to wild type fruit. Refer Supplementary Fig. 1 for SGA pathway intermediates. Knockout of GAME36 resulted in significant accumulation of precursor SGAs (i.e., hydroxytomatine, hydroxy-dehydrotomatine, hydroxytomatine isomers) with almost complete loss of downstream SGAs (acetoxytomatine, acetoxy-hydroxytomatine and Esculeoside A) in tomato fruit tissues. Notably, hydroxylated SGAs are the most abundant SGAs in red fruit of game36 mutant lines compared to the wild type red fruit. The values indicate mean of biological replicates ± standard error mean (n = 4 for wild type, n = 3 for #14 and #18 lines and n = 2 for #41 and #50 lines). Asterisks indicate significant changes compared to wild type samples as calculated by a two-tailed Student’s t-test (*P-value < 0.05; **P-value < 0.01; ***P-value < 0.001). LC-MS was used for targeted SGAs profiling. Source data (panel a, c, d) are provided as a Source Data file.