OBTAINING AND REGENERATION OF STEM CELL LINES WITH ENHANCED BETULINIC ACID FROM ENDEMIC Rosa pisiformis (ROSE) PLANT

Project no. 23788 · Status: Kapanmış (Yayın bekleniyor) · 2022

Summary

The fact that stem cells are totipotent indicates that they contain all genetic information. Therefore, the components or chemicals obtained from stem cells offer the highest quality and most efficient products. The reasons why anticancer drugs used in the pharmaceutical sector are expensive are that the chemicals in these drugs are contained in plants at very low levels and that high-cost extraction processes are performed with plants collected from nature. However, cultured plant stem cell applications offer a production system that does not harm nature and is sustainable, provides uniform quality and efficiency. In addition to its pharmacological effects such as anti-cancer, anti-inflammatory and anti-HIV, betulinic acid also provides collagen increase and the fact that Rosa damascena species (rose) is also important in our country's exports has emphasized its importance in the cosmetic industry and made this plant secondary metabolite a high-value product. Plants are capable of producing a wide variety of secondary metabolites with very wide functions used for medical purposes; for example, betulinic acid and its derivative betulin ester with diaminobutyl acid (BE-Dab-NH2 ) and betulin are natural compounds found in Rosa spp. fruit that are effective on collagen synthesis. They are found in very low concentrations in the plant (Wenzig et al., 2008). Since their chemical synthesis via the complex betulinic acid pathway does not have a commercial value; alternative biotechnological sources and plant cell cultures containing heterologous expression systems have been developed.In this thesis study, it was planned to increase the betulinic acid content of Rosa pisiformis, a rose species that is endemic and not previously studied in our country among the Rosa species spread in our country. Following the content amount determination, Rosa pisiformis root cells will be taken into suspension culture with micropropagation studies and the continuity of the cells will be ensured for the studies within the scope of the project. Studies to increase the betulinic acid accumulation in the cells will be initiated. As a result of the inhibition of the PAL (phenyl alanine ammonium lyase) pathway based on the betulinic acid pathway, more terpenes and therefore the accumulation of betulinic acid, a lupane type triterpenoid, is increased. It was also aimed to increase the betulinic acid amount in Rosa pisiformis root cells by using AOPP (alpha-aminooxy beta-phenyl propionic acid), a strong phenylalanine analogue, for the inhibition of the PAL pathway. Mutagenesis studies will be conducted for the permanence of cell lines. It is planned to use EMS (ethyl methanesulfonate) chemical in mutagenicity tests because of its high mutagenic effect, small DNA deletions and additions and high efficiency. A plating system platform will be created for the selection of cell lines. The selection of very sensitive stem cells will be carried out quickly and with this highly efficient system. These cell lines, which are thought to have increased betulinic acid accumulation, will be proven by the HPLC (high performance liquid chromatography) method to be performed and their quantitative content will be determined and the results will be presented. In addition, Rosa plants with high betulinic acid content can be produced by regenerating stem cell lines with increased betulinic acid amounts. Wenzig, E.M., Widowitz, U., Kunert, O., Chrubasik, S., Bucar, F., Knauder, E. ve Bauer, R. 2008. Phytochemical composition and in vitro pharmacological activity of two rose hip (Rosa canina L.) preparations. Phytomedicine; 826-835

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