Fig. 8

Flow chart showing four subsequent steps for the development of protocol for an early selection and regeneration of carrot somatic hybrids. (1.1) protoplast fluorescence after staining with rhodamine B isothiocyanate (RBITC); (1.2) protoplast fluorescence after staining with fluorescein diacetate (FDA); (2.1) fluorescence of FDA- and (2.2) RBITC- stained protoplasts as well (2.3) dual-colour fluorescence observed in the post-fusion protoplast mixture (putative hybrid cells are marked by arrows); (3.1) a schematic representation of the developed thin alginate feeder layer culture system; (4.1–4.3) a micromanipulator-based (4.1, 4.3) manual selection of dual-labelled hybrid cells (4.2); (4.4) development of somatic embryos observed in microcallus cultures derived from selected putative hybrid cells; (4.5) acclimatized to ex vitro conditions ‘Dolanka’ ( +) D. carota subsp. gadecaei hybrid. Scale bar: 100 μm