NOVEL TECHNIQUES FOR GROWTH-CHARACTERIZATION AND PHYTOCHEMICAL ANALYSIS OF PLANT-CELL SUSPENSION-CULTURES

Major problems associated with plant cell suspension cultures are the variability and instability of the cultures. Therefore, characterization of a suspension culture in every experiment is necessary to allow a comparison between different experiments. For this purpose, some new techniques were developed. A technique for growth characterization has been developed based on the dissimilation of the cultures. The main advantages of this method are that it is non-destructive, it supplies a dissimilation curve for each individual Bask, and it is accurate. Here we present an accurate calculation for the curves and the most accurate way to measure them. Another technique was developed for the rapid quantification of the main organic intracellular components (especially carbohydrates and amino acids). Aqueous extracts are prepared and the components are quantified by H-1-nmr spectroscopy. The main advantages of this method are nonselectivity and good sensitivity. The developed techniques were applied in an experiment in which cultures growing on media containing different nitrogen levels were compared. The optimum quantity of nitrogen for growth and alkaloid production was found to be half the amount normally present in Murashige-Skoog medium.