OPTIMIZATION OF CONDITIONS OF CULTIVATION FOR SUSPENSION CULTURE OF THE RHODIOLA ROSEA L.

Issue: 
9
Year: 
2016

T.A. Savina
Ph.D. (Biol.), Head of Biotechnology Section, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow)
E-mail: savina-tatyana57 @yandex.ru
S.B. Myasnikova
Research Scientist, Biotechnology Section, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow)
E-mail: vilarnii@mail.ru
M.V. Balakina
Ph.D. (Pharm.), Leading Research Scientist, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow)
E-mail: m_balakina@mail.ru

In the laboratory of biotechnology VILAR is carried out the cultivation of suspension cultures of Rhodiola rosea (collection number 04868244-012-2014). The aim of this study is to optimize the composition of nutrient medium for the cultivation of suspension cultures of Rhodiola rosea in vitro. The main stage of investigation consisted in studying impact on growth and biosynthetic properties of suspension culture of Rhodiola rosea, groups of macronutrients, are part of the nutrient medium according to the recipe Murashige and media in the form of salts (NH4NO3, KNO3, MgSO4 × 7H2O, KH2PO4). The main stage of investigation consisted in studying impact on growth and biosynthetic properties of suspension culture, group of the macrosalts which are a part of a nutrient medium: nitrogen (N), phosphorus (P), calcium (Ca), magnesium (Mg), potassium (K). Were studied four concentrations of the total content of macro-elements (nitrogen (N), phosphorus (P), calcium (Ca), magnesium (Mg), potassium (K)) in the composition of the nutrient medium. The results of the experiment showed that increasing the concentration of macrosalts twice had no effect on growth parameters of culture. The decrease in the concentration of macrosalts in the nutrient substrate in two times led to some decrease of biomass growth, it was only 9%. The highest salt concentration caused a dramatic decrease of biomass growth, and it was 34%. Chemical analysis of cell mass of Rhodiola rosea was carried out using method of spectrophotometry. The decrease in the total concentration of macrosalts leads to a decrease in the biosynthetic activity of the culture is 6%. It was established that for growth and biosynthetic activity of suspension cultures of Rhodiola rosea it is possible to vary the total content of macrosalts within a two-fold increase. This is a fairly significant step in quantitative terms. In continuation of the experiment on optimization of the growth conditions of the cell culture of Rhodiola rosea conducted a study in which they took a triangular matrix method, was varied with the ratio between the nutrients phosphorus, calcium and magnesium at their unchanging total concentration. It is established that the growth of suspension cultures of Rhodiola rosea is provided by a predominance of phosphorus in the nutrient medium. The growth of biomass is increased by 5%. The increasing of amount of phosphorus in the nutrient medium significantly influenced on the accumulation of phenylpropanoids (2,67%). In the synthesis of phenylpropanoids the role of calcium and magnesium in the composition of the nutrient medium are increased (3,2 % and 3.07 of phenylpropanoids). At the HPLC chromatograms was observed by the presence of a peak rosavine with time ~ 4 minutes, but the peak response was negligible. On the TLC plates for all four samples revealed a spot with Rf=0,514-0,563. The spot of rosavin at TLC-plates not detected, that may indicate at slight rosavin content in the biomass samples.As a result of experiments, it was found that the growth and biosynthetic activity of the suspension culture Rhodiola rosea it is possible to vary the content of the sum of macrosalts doubling. On the basis of obtained data were calculated the optimal ratio between the three subjects nutrients (phosphorus, calcium, magnesium) required for tissue growth for the synthesis of phenylpropanoids.

Keywords: 
Key words: cell strain of Rhodiola rosea
a nutrient medium
culturing conditions (conditions of cultivation)
makrosalts
rosavin
phenylpropanoids
HPLC
spectrophotometry.