In vitro regeneration of transgenic plants via somatic embryogenesis in Pelargonium peltatum and via organogenesis in P. zonale. (a) P. peltatum WT plants. (b) P. peltatum inflorescence. (c) P. zonale WT plants. (d) P. zonale inflorescence. (e-f). Production of somatic embryos in a callus of P. peltatum cultivated in selective Morphogenesis Induction Medium (MIM). (g). P. peltatum developing embryos in selective Elongation Medium (EM). (h). P. peltatum transgenic plantlets in Rooting Medium (RM). (i-j). Adventitious buds in a callus of P. zonale in selective MIM. (k). P. zonale shoot elongation in selective EM. (l). P. zonale rooted plantlet in RM. (m-n). Detection of transformation events in both Pelargonium spp., GFP green fluorescence is clearly visible in the initial whitish callus with a disorganized growth. (o-p). Chlorophyll shows strong red autofluorescence that could mask the green fluorescence of transformed cells, it becomes increasingly difficult to identify in the subsequent organogenic callus and in the adventitious buds. (q-r). Green fluorescence was observed in regenerated shoots but is masked by the chlorophyll in the young leaves. (s-t). Green fluorescence can be observed in the periphery of young leaves where chlorophyll does not accumulate. (u-v). Green fluorescence is especially evident in the roots, where the chlorophyll is absent. In general, within the same organ, GFP detection varied in different tissues or cell types depending on their chlorophyll content.
García-Sogo et al. BMC Plant Biology 2012 12:156 doi:10.1186/1471-2229-12-156