OBTAINING CELL CULTURES OF CICHORIUM INTYBUS L. IN VITRO, AS A SOURCE OF INULIN

Abstract

The results of cultivation of Cichoriumintybus L. in culture in vitro are presented. It is shown that the processes of morphogenesis are significantly influenced by growing conditions, in particular, the hormonal and mineral composition of the nutrient medium, as well as lighting modes. Callus tissue was obtained from leaf explants isolated from C. intybus plants. Leaf segments were cultivated on a nutrient medium containing mineral salts according to the MS prescription, as well as auxins - indolyl-3-acetic (IAA), naphthylacetic (NAA) and 2,4-dichlorophenoxyacetic (2,4-D) acids at a concentration of 5.5 - 9.5 mg / l in combination with cytokinin 6-benzylaminopurine (BAP) - 2 mg / l. The formed tissue was transplanted onto a fresh nutrient medium every four weeks, taking into account the color and consistency. Callus growth was determined by weighing in a laminar box at the beginning and end of the passage (mg). Callus tissue was cultured on agar nutrient medium in Petri dishes. Then they were placed in opaque grow tents with radiation aligned with the flux density of photosynthetic photons and different ratios of its levels in the region of 660 nm (R - red) and 730 nm (FR - far red). The control variant was placed in a light room, where illumination was created with white linear fluorescent lamps with an intensity of 150 µmol/m2s. At the end of the growing cycle, the color, texture of the callus and its growth were taken into account.  It is shown that the process of callus formation depends on the auxin used and its concentration. The maximum increase in callus tissue was obtained on an MS medium containing IAA or NAA at a concentration of 8.5 mg/l. In the presence of NAA, a non–morphogenic callus was formed, and when using IAA, a morphogenic callus was formed. Studies have shown that the use of various lighting conditions (FR>R, FR=R, FR<R) have a significant stimulating effect on the growth of callus tissue, as well as on the accumulation of inulin in it. Moreover, the maximum value of inulin in the callus tissue (7.55-7.95%) was obtained in the FR>R variant, as well as on a nutrient medium containing IAA.