Figure 4.

Chlorogenic acid localization in embryogenic cell clusters (C3) and during embryo redifferentiation after 1 day (R2), 3 days (R3), 10 days (R4) and 3 weeks (E1) in redifferentiation medium after cell density reduction. Protuberance of newly generated embryos emerge after 10 days in redifferentiation medium and globular-shaped embryos are obtained after 3 weeks. For chlorogenic acid localization, fresh samples were mounted on a glass slide in a drop of water and observed using a Zeiss LSM880 multiphoton microscope, equipped with a Chameleon Ultra II laser. Excitation wavelength was set to 720 nm and emission was observed with a band-pass filter 386–502 nm (blue). The presence and accumulation of total chlorogenic acids during embryo redifferentiation was confirmed by multiphoton microscopy combined with emission spectral analysis of total chlorogenic acids (Figure S2). The scale bar is set to 100 µm.