Changes in bioactive components of Aristolochia tagala .Cham, a rare species of medicinal importance during its in vitro development through direct regeneration

Abstract

Tissue culture propagation system was developed for Aristolochia tagala, a threatened medicinal plant, using apical bud explants. The most effective medium was found to be MS medium supplemented with BAP (3 M), KIN (0.5 M) and activated charcoal (0.1%). The addition of activated charcoal helped in circumventing the problem of polyphenol exudation from the explants which hampered the regeneration of adventitious shoots. A maximum of 12.6 shoots were obtained on average from the apical bud explants after 25 days of inoculation. Well developed shoots were rooted on MS medium supplemented with indole acetic acid (1.5 M), Kinetin (1.5 M) and 6-benzylaminopurine (0.5 M). Regenerated shoots from the apical buds were successfully rooted and acclimated to greenhouse conditions. Qualitative and quantitative analysis of bioactive compounds was done at various stages of development, so as establish the effect of culture conditions on the production of bioactive components. Comparisons were made between three types of plant material from the same clone: leaves from field-grown plant, leaves from in vitro apical bud cultures and leaf derived callus. It was observed that the leaf derived callus showed the presence of components which were not there in the in vivo leaves, suggesting the influence of in vitro developmental conditions.\n\nTagalaSummary Statement\n\nThis is the first report on direct regeneration of Aristolochia tagala, (a threatened yet important medicinal plant) using apical bud explants derived from mature plants. Also, for the very first time we report the results of analysis of secondary metabolites during its various stages of in vitro development.\n\nAbbreviationsAC - activated charcoal; BAP - 6-benzylaminopurine; 2,4-D - 2,4- dichloro phenoxyacetic acid, IAA - indole acetic acid, IBA - indolebutyric acid, KIN- kinetin, MS- Murashige and Skoog (1962); NAA- -naphthaleneacetic acid.