Dormancy breaking, indirect somatic embryogenesis, and encapsulation of somatic embryos in Black Zira

Black Zira ( Bunium persicum ) is an important endemic medicinal plant in Iran. This research was conducted out to assay breaking seed dormancy, develop new protocols for callus induction, somatic embryogenesis, and artificial seed. Seeds were treated with ultrasonic waves in different periods (0, 1, 3, 5, 7, and 9 min). Then, two experiments with IBA (0, 1, 2, 3, and 4 mg L − 1 ) and a combination of different levels of 2,4-D (0, 1, 2, and 4 mg/l) and BAP (0, 0.5, 1, and 2 mg/l) were designed. Different concentrations of sodium alginate (2, 3, and 4%) and CaCl 2 (50, 75, and 100 mM) were prepared for artificial seed production. According to the results, the 7 min ultrasonic waves had the maximum effect on germination (81.87%), while the highest negative effect on germination (2.5%) was reported from 9 min exposure time. In the IBA experiment, the highest percentages of callogenesis, callus weight, and adventitious root were obtained from MS medium containing 4 mg/l IBA. Moreover, 1 mg/l IBA resulted in the highest embryogenesis percentage. In the second experiment, the highest percentage of callus formation and callus weight were achieved in 1 mg/l BAP + 4 mg/l 2,4-D, 0.5 mg/l BAP + 4 mg/l 2,4-D, and 2 mg/l 2,4-D media. The maximum embryogenesis percentage was seen in 0.5 mg/l BAP + 1 mg/l 2,4-D, 0.5 mg/l BAP + 2mg/l 2,4-D, and 1 mg/l 2,4-D media. The highest germination rate (77.77%) was obtained from encapsulated embryos with 3% sodium alginate and 100 mM CaCl 2 .