(a) Typical phenotypes of vector control (VC) and 35Sp-LeTYR/35Sp-MjDOD line no. 3 (TYR/DOD no. 3) calli after 4- and 8-weeks subculture. (b) Change in betaxanthin concentration over time in transgenic calli line no. 3 after subculture. Betaxanthins were extracted in 80% (v/v) methanol containing 0.04% (v/v) HCl, and concentration was calculated from absorbance at 480 nm. (c) Identification of betaxanthins by LC-TOF-MS in 35Sp-LeTYR/35Sp-MjDOD transgenic BY2 cells. Chromatography profiles of extracts from vector control (dashed line) and 35Sp-LeTYR/35Sp-MjDOD line no.3 (thick line). Peak 1, retention time 23.8 min, m/z = 340.114, λmax = 470 nm, Glutamine-Bx (Vulgaxanthin I); Peak 2, 27.4 min, m/z = 313.104, λmax = 470 nm, Threonine-Bx; Peak 3, 41.3 min, m/z = 309.1084, λmax = 480 nm, Proline-Bx (Indicaxanthin).