FIELD: biotechnology.
SUBSTANCE: method for obtaining the substance from callus cultures of St. John's wort (Hypéricum perforatum L.) relates to the field of biotechnology. The method is carried out by stepwise sterilization of the seeds of St. John's wort, for which plant explants are initially treated in a soapy solution, then washed with distilled water, then placed in a laminar box in 70% alcohol for 1 minute, and then into the main sterilizing solution, which is whiteness 50% or 100% when exposed for 10-20 minutes, or hydrogen peroxide 36% when exposed for 15 minutes, or sublimate 0.1% when exposed for no more than 15 minutes. Next, the seeds are washed with autoclaved distilled water three times for 15 minutes and the plant explants are placed on the Murashige and Skoog nutrient medium without the use of phytohormones to obtain seedlings, cultivated in a thermostat at a temperature of 24°C. The resulting seedlings are cut under sterile conditions and placed on a Murashige and Skoog nutrient medium with the addition of phytohormones 0.2 mg/l indoleacetic acid and 0.5 mg/l kinetin to induce callusogenesis. After 28-30 days, a callus culture is obtained, which is dried at a temperature not exceeding 40°C and then crushed. Then an extract is prepared by dissolving 1 part of the callus culture in two parts of 40% alcohol, mixed and centrifuged for 15 minutes at a speed of 10,000 revolutions, followed by pouring the liquid into an empty sterile tube, repeat the procedure and pouring the liquid into the same tube, pour 40% alcohol again, mix and put in a refrigerator to settle for a day at a temperature of 4°C, then centrifuge again, pour the liquid into the same test tube and dry the resulting extract.
EFFECT: method allows to obtain a substance with antioxidant and antibacterial properties, including against Escherichia coli and Staphylococcus aureus, by cultivating plant cells and tissues of Hypéricum perforatum L., and the process of obtaining a callus culture is carried out in laboratory conditions and is not associated with growing and collecting plants in field.
1 cl, 2 dwg, 5 ex
