Influence of plating density, sucrose and light during development on the germination and vigour of Medicago sativa L. somatic embryos after desiccation

Somatic embryo quality is an important aspect of artificial or synthetic seed production and is dependent on proper morphological development and deposition of sufficient storage reserves to support early growth of the seedling. The abilities of a somatic embryo to survive desiccation and to establish a rapidly growing seedling were used as estimates of quality and were dependent on a number of factors during the elongation and maturation phases of somatic embryo development. Embryo quality was significantly enhanced when the sucrose content of the elongation and maturation media was increased to 50 g I−1 from 30 g I−1. This increased dry weight to >2 mg embryo−1 suggesting greater deposition of storage reserves. Higher sucrose concentrations were inhibitory and reduced survival after desiccation and seedling vigour. An optimum number of embryos can be supported on one Petri plate; the number was controlled by the plating density of the embryogenic cell clusters spread on the elongation medium. This optimum was dependent to some extent on the sucrose content of the medium, indicating that sucrose had a nutritive effect on embryo development and not solely an osmotic effect. Light intensity was another critical factor. Intensities > 75 μmol m−2 s−1 photosynthetic photon flux density (PPFD) at the elongation stage of development blocked globular-stage somatic embryo development. Maturing somatic embryos were far less sensitive to high light intensities and development was more rapid at 150 μmol m−2 s−1 PPFD. Light quality was manipulated by covering plates with filters that selectively transmitted certain regions of the visible spectrum. All filter treatments reduced embryo yield and quality compared with embryos developing under fullspectrum light; varying the light spectrum does not seem to be a viable method of enhancing somatic embryo quality.