FIELD: biotechnology.
SUBSTANCE: invention relates to the field of biotechnology. Invention is a method for obtaining a root culture in vitro Potentilla alba L. – a flavonoid producer, characterized in that the Potentilla alba L. seeds are degreased, dried, sterilized, thoroughly washed with sterile distilled water, is placed on medium AS (Straits), sprouted in darkness, transferred into separate containers with medium of the same composition and placed on light until formation of young leaves (explants plants), separated from roots aboveground part of seedlings, leaves, subfamily knee and hypocotyl are cut into segments with size of 1.0–1.5 cm, pierced with an insulin syringe needle along the vein along the leaf, along the epicotyl and hypocotyl, to the vascular system of plants, wild (non-modified) strain Agrobacterium rhizogenes (A 4) is grown on nutrient medium YEV for 48 hours in darkness at temperature 26 °C, or at temperature of 32 °C for 24 hours on rocker with circular motion, prepared plant explants are transferred onto YEV medium with extended agrobacteria, held in magnetic bath for 10–100 seconds and incubated for 12–24 hours, are washed with sterile nutrient medium MS ½ N, transferred to nutritious agarized medium Murashige and Skoog MS ½ N with addition of klaforan (250 mg/0.5 l) for elimination of A. Rhizogenes (A 4), is exposed to a "light" room until formation of transformed roots, the roots are transplanted into a fresh nutrient medium of Gamborg B-5 without hormones for complete elimination of bacteria A. Rhizogenes (A 4) and growing in a "dark" room in swinging conditions.
EFFECT: invention provides constant high rate of growth of isolated roots of Potentilla alba L on hormone-free nutrient media with a high level of flavonoid biosynthesis.
5 cl, 3 dwg, 1 tbl, 3 ex
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Authors
Dates
2020-02-14—Published
2019-03-22—Filed