Production of Medicinally Important Metabolites by Rauwolfia Cell Cultures

Abstract

During the present investigations nodal explants of Rauwolfia serpentina were inoculated on MS nutritional medium supplemented with different concentrations of various growth regulators to see their effect on growth and development. Our past and present studies have revealed that MS is a suitable medium for the culture of this plant species and does not require addition of other complicated organic addenda. Table1-6 and figures 1-4 deal with multiple shoot formation on the nodal segments under the influence of different growth harmones. A large number of multiple shoots were induced on MS supplemented with 0.5 mg/1 each of BAP and 2, 4-D. These shoots were severed from the parent axis and induced to root under the influence of various treatments. These findings have been detailed in figure 5-9 and table 7. These plantlets after induction of roots were transferred to soil, where they exhibiting normal growth. Under the influence of a different set of harmones the nodal segments have been induced to callus. A massive and healthy callus was induced on MS supplemented with 1.5 mg/1 on kinetin and 1.0 mg/1 of 2, 4-D (Fig.11). This callus could be easily maintained and multiplied further and will be used for the isolation of alkaloids in the future. This callus will also be tested for its regenerating capabilities. Another kind of callus was induced when MS was supplemented with 2.0 mg/1 of BAP and 1.5 mg/1 of IAA (Fig .18). This callus was granular and gave rise to embryoid structures which afterwards produced normal plants (Figs. 21, 22 and 23). These plantless were allowed to attain a reasonable length and after acclamatization (Fig.28) were transferred to soil conditions (Fig.29). These plants are healthy and have established themselves to the natural conditions. Copious root formation has also been induced using different hormonal combinations. These results have been supported by figures 24, 25 and 26. These roots will be further multiplied and utilized for the extraction of the medicinally important metabolites. As a result of present investigation numerous shoots could be induced on the nodal segments, rooted and then transferred to the field after acclimatization. Furthermore, copious callus has been induced which could be maintained undifferentiated condition or made to form either roots or shoots for propagation or alkaloid analysis. The induced roots could also be kept in undifferentiated satate for an indefinite culture period without any obvious loss in its vigour. Similarly the callus and the shots could be maintained in undifferentiated satate for either organogenesis or alkaloid synthesis. All-important Rauwolfia alkaloids have been isolated from the regenerated roots and shoots. Although active metabolises have also been isolated from callus cultures as well, their concentration was not detected in appreciable quantities. A comparison of the present results with our own previous results and those of others indicated that the two important alkaloids viz. Reserpine and serpentine were also synthesized in appreciable amounts by these cultures. These results thus indicate the economic importance of this biotechnology which could possibly be exploited on a large scale. Presence of other fluorescent bands deserves identification and confirmation which might add to the economic importance of this technology