Typical examples of metabolite differential levels among the ancestral and evolved lines. (a) Sections of chromatograms showing orotate or orotic acid (denoted by an arrow) abundance among all the lines. (b) Mass spectrum of orotate purified standard and mass spectrum of the identified peak as orotate in both strains. (c) Box and Whisker plots of metabolites involved in nucleotide related pathways. 1 and 3 represent MG and DH lines (ancestors); 2 and 4 represent MGAdp and DHAdp lines (evolved). The top and bottom of each box represent the 25th and 75th percentiles, the centre square indicates the mean, and the extents of the whiskers show the extent of the data. For each metabolite, the maximal measured peak area was normalized to a value of 100.