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SECONDARY PRODUCTS OF CULTURED PLANT CELLS
OBTAINING OF HIGHLY PRODUCTIVEARNEBIA EUCHROMA CELL
LINE-THE SOURCEOFSHIKONIN
O.V.Zakhlenjuk,O.O.Poronnyk, V.A .Kunakh
Institute ofMolecular Biology and GeneticsNAS Ukraine, 252143 Kyiv, Zabolotnogostr.,
150,
As a starting material was used the AE-1 strain o f Amebia euchroma
suspension cell culture (the registration number is BCKKBP N 42). The
strain can accumulate at 14th day o f maintenance between 0.8 and 2.0%
shikonin and its derivatives as calculated per dry biomass. Studies were
performed along the two routes: the selection of highly productive somaclonal
variants and the manipulation with the growing conditions to support higher
productivity o f the cultured cells. Various selection strategies were used:
the mass selection, the cloning of the cell aggregates, the selection of differing
fractions of culture, the selection by the intensity and the performance of the
tissue coloration during maintenance on different nutrient media.
The best results were produced by means o f time to time selection of
the m inor extensively stained cell aggregates 1 to 2 mm size concomitant
with the selection of cultures to exhibit pink coloration o f the tissue and
nutrient medium. Whereas the starting experiments showed variations
between the flasks from 0.2 to 1.4% then the final ones resulted in 2.9 to
8.4% per dry tissue mass. Superior cell line productivity by shikonin amounted
to 1.6 g/l of nutrient medium over 14 days of growth or 114.2 mg/l of shikonin
per day.
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