Fig. 4From: A fast and simple LC-MS-based characterization of the flavonoid biosynthesis pathway for few seed(ling)sReduction of seed material and proof of principle with seeds. Reduction of seed material down to one seed (a,b) and application to a set of selected mutants (c-e) with extracted non-hydrolysed, extracted hydrolysed and non-extracted hydrolysed samples analysed by LC-ESI-MS-QTRAP(MRM). a Flavonoids with detectable and quantifiable responses in 50, 25, 10, 5 and 1 A. thaliana Col-0 seed(s). Grey: LOD but not LOQ is passed, black: LOQ is passed for the majority of replicates. See also Additional file 6: Figure S9 and Additional file 5: Figure S7 and S8 (effect of hydrolysis on analysed flavonoids). b Responses normalized with the respective deuterated internal standard. Several scales are used for each subfigure. Grey values of scales correspond to the grey values of the substances. Boxed: optimal seed number according to this experiment. Error bars = STDEV. (Additional file 6: Table S11: statistics) c-e) Mutant analysis: Heatmaps showing log2 of fold differences relative to the respective wild types. d Same as in (c) and (e) for tt7-1 with appropriate scales. e Same data as in (c) but normalized with kaempferol. -: LOD not passed, #: LOD but not LOQ passed, +: LOQ passed for the mutant but not for the wild type. Lines in grey shades group the three sets of mutants: enzymes, TTG1-MBW complex components, light signalling mutants. Additional file 7: Tables S12 and S13: statistics. Additional file 7: Figure S10: responses normalized with respective deuterated internal standard. c Detected catechin which is most likely derived from epimerization of epicatechin (see Fig. 3)Back to article page