Seed dormancy breaking of an endangered medicinal tree species (Taxus baccata L.) using embryo culture

Abstract

Natural regeneration of Taxus baccata L. is constrained due to the depth of seed dormancy requirements (often taking two or more years) and low seed germination. Further, the conventional method of vegetative propagation by cuttings is associated with difficulties in rooting. Hence, for the first time, this study describes an efficient and reproducible in vitro protocol for breaking the dormancy of seeds from the endangered forest tree T. baccata L. via zygotic embryo culture. Embryos isolated from 100% sterile seeds were cultured on DCR medium that contains sucrose (30 g/l), agar (8 g/l), and activated charcoal (5 g/l), fortified with different concentrations of Plant Growth Regulators (PGRs), and held at a temperature of 25 ± 2 ºC in a growth room. The results revealed that the in vitro embryo germination percentage was mostly affected by gibberellic acid (GA3) and thidiazuron (TDZ). Among the nine treatments, the treatments with 0.5 mg/l TDZ and 1 mg/l GA3 showed the highest germination (100%), while the other treatments all increased the germination percentages significantly compared to the control (37.5%). The 1/2 DCR medium with the addition of 0.1 mg/l indole-3-butyric acid (IBA) resulted in the highest rooting ratio (94%). However, the greatest root and hypocotyl elongation (59.37 ± 3.77 and 62.75 ± 4.43 mm, respectively) occurred when seedlings were cultured on 1/2 DCR medium containing 0.5 mg/l BA. Plantlets were transplanted into plastic pots containing an autoclaved garden soil, sand, and vermiculite mixture (1:1:1) and held at a temperature of 25 ± 2 ºC in a growth room for 4 weeks before being transplanted into the greenhouse. These results indicated that the protocol developed during the current study will be useful to overcome seed dormancy and for multiplication and conservation of the species T. baccata L.