GC-MS PROFILE OF IN VIVO, IN VITRO AND FUNGAL ELICITED IN VITRO LEAVES OF HYBANTHUS ENNEASPERMUS (L.) F. MUELL
Objectives: Investigation of the bioactive compounds from the methanol leaf extracts of in vivo, in vitro and fungal elicited in vitro plants of Hybanthus enneaspermus through GC-MS analysis.
Methods: The leaf explants were cultured on MS (Murashige and Skoog) medium supplemented with different concentrations of NAA (1-naphthalene acetic acid) for callus induction. The calli was treated with four different fungal elicitors, namely, Mucor prayagensis, Trichoderma viride, Fusarium moniliformis and Aspergillus niger on suspension culture containing the same growth regulators for two weeks. Regeneration of shoots was achieved from the fungal treated and untreated calli on MS medium fortified with different concentrations of cytokinins. Rooting was achieved from the isolated shoots on half strength MS medium containing different concentrations of auxins. The phytochemical composition was analyzed from the methanol extracts of in vivo, in vitro and fungal treated leaves of in vitro plants using gas chromatography and mass spectroscopy (GC-MS).
Results: Of the different concentrations and combinations of NAA, well developed green compact reproducible calli were obtained on MS medium supplemented with 10 Âµmol NAA+4 Âµmol BAP (6-benzylaminopurine). Shoots were regenerated from the fungal treated and untreated calli on MS medium containing 10 Âµmol BAP+6 Âµmol KIN (kinetin-6-furfurylaminopurine). Rooting was achieved on half strength MS medium supplemented with 9 Âµmol NAA. The GC-MS analysis revealed that the leaves of in vivo and in vitro plants contained 16 different phytochemicals, whereas, the fungal treated in vitro plants showed more number of phytochemicals, i.e., 22 (Mucor prayagensis), 26 (Trichoderma viride), 19 (Fusarium moniliformis) and 21 (Aspergillus niger) compounds.
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