METHOD FOR MAKING CALLUS CULTURE OF DRACOCEPHALUM PALMATUM STEPH. IN VITRO CONDITIONS Russian patent published in 2020 - IPC C12N5/04 

FIELD: biotechnology.

SUBSTANCE: invention represents a method for obtaining a callus culture of cells of a wild-growing plant of Dracocephalum palmatum Steph. in vitro, involving sterilization of the Dracocephalum palmatum Steph. seeds with solution of 3 % hydrogen peroxide for 5 minutes, 80 % ethyl alcohol for 1 minute, three-time rinsing in sterile distilled water, placing sterile seeds on a solid nutrient medium without hormones of the following composition, mg/l: NH₄NO_{3 –} 33000, KNO_{3 –} 38000, CaCl₂×2H₂O – 8800, MgSO₄×7H₂O – 7400, KH₂PO₄ – 3400, KI – 166, H₃BO₃ – 1240, MnSO₄×4H₂O – 4460, ZnSO₄×7H₂O – 1720, Na₂MoO₄×2H₂O – 50, CuSO₄×5H₂O – 5, CoCl₂×6H₂O – 5, FeSO₄×7H₂O – 5560, Na-EDTA – 7460, mesoinositol – 100, nicotinic acid – 100, pyridoxine – 100, thiamine – 100, saccharose – 2500, water – 1000 ml, agar – 7000. Further, leaf explants obtained from seedlings are placed in a nutrient medium of the following composition, mg/l: NH₄NO₃ – 33000, KNO₃ – 38000, CaCl₂×2H₂O – 8800, MgSO₄×7H₂O – 7400, KH₂PO₄ – 3400, KI – 166, H₃BO₃ – 1240, MnSO₄×4H₂O – 4460, ZnSO₄×7H₂O – 1720, Na₂MoO₄×2H₂O – 50, CuSO₄×5H₂O – 5, CoCl₂×6H₂O – 5, FeSO₄×7H₂O – 5560, Na-EDTA – 7460, mesoinositol – 100, nicotinic acid – 100, pyridoxine – 100, thiamine – 100, saccharose – 3000, casein hydrolyzate – 500, kinetin – 1, 2,4-dichlorophenoxy acetic acid – 1, water – 1000 ml, agar – 12000. Cultivation of plants is carried out in the dark, at temperature of 26±1 °C, humidity of room is 70±5 %, the subcultivation cycle is 4 weeks.

EFFECT: invention makes it possible to produce callus culture of Dracocephalum palmatum Steph_ under in vitro conditions, which can be used as a source of flavonoids of therapeutic action.

1 cl, 3 tbl