Establishment of grapevine embryogenic liquid culture and induced somatic embryogenesis
Somatic embryogenesis is a process in which somatic cells can be used to regenerate embryos and whole plants. Embryogenic cell suspensions of 'Chardonnay' and 'Richter 110' grapevine cultivars were established in 8 different liquid media, cultures had been later maintained in solid media. By the end of the sixth week, we achieved 7-fold increase of fresh weight in the case of the 'Chardonnay' cultivar, using MSM1 and CP2 liquid media. Four different pH values of hormone-free MSM1 media were tested in our experiments (pH 5.8, pH 4.6, pH 4.3 were adjusted before autoclaving; or pH 4.3 after autoclaving). In the case of pH 5.8, we measured a 16-fold increase of fresh weight of the 'Richter110' cultivar. Various culture densities (0.25-4 mg mL-1) were tested, and found that the one with 2 mg mL-1 density was the most efficient. Somatic embryogenesis proved to be significantly more efficient when the medium was supplemented with 0.5 g L-1 MES buffer [2-(N-morpholino) ethanesulfonic acid]. In this case, however, the embryos regenerated developed highly elongated hypocotyls, which inhibited the germination of the embryos as well as the successful regeneration of plants. In a half-strength MSM1 medium with a cell density of 1 mg mL-1, the cultures were composed of well synchronised globular stage embryos. In this setup, however, further differentiation was reversibly inhibited. This inhibition can be resolved by a second dilution of the cultures (to 1 mg mL-1 cell density). When hormone-free MSM1 medium was used after the second dilution, which was enriched with 1 g L-1 activated charcoal, synchronised and well developed cotyledonary stage embryos were differentiated in the liquid cultures by the sixth week.
Forgács, I., Suller, B., Zok, A., Pedryc, A., Oláh, R., Deák, T., Bisztray, Gy.D. and Szegedi, E. (2017). Establishment of grapevine embryogenic liquid culture and induced somatic embryogenesis. Acta Hortic. 1157, 113-118
Vitis vinifera, activated charcoal, cell suspension, maltose, 2-naphtoxyacetic acid, plant regeneration