Abstract
The study illustrates the system for enhanced production of a medicinally important unexplored compound, carvone occurring naturally in Anethum graveolens. The effect of salicylic acid (SA) on biomass yield, carvone biosynthesis, growth and major enzymatic antioxidant parameters in A. graveolens was evaluated. The effects of different combinations of benzyl adenine (BA) and 1-Naphthalene acetic acid (NAA) were tested. Murashige and Skoog (MS) medium comprising 1.76 µM BA + 3.24 µM NAA was the best for friable callus induction. The friable callus was used for the initiation of cell suspension culture. MS salts in combination with 4.4 µM BA and 2.6 µM NAA, 3% sucrose was appropriate for cell growth and bioactive compound accumulation. The cell suspension cultures were then treated with SA (0.1, 0.75 and 1.5 mM) as an elicitor for four weeks. An up regulation of enzymatic antioxidants, ascorbate peroxidase (APX); superoxide dismutase (SOD) and catalase (CAT) activity with increasing concentrations of SA whereas a reduction in guaiacol peroxidase (GPX) activity was recorded at the end of the growth phase. The results also showed that higher concentrations of SA significantly increased malondialdehyde (MDA) and Proline content. Cell suspension culture was then subjected to extraction and isolation. The quantification of carvone through HPLC analysis revealed highest amount of carvone (0.063%) in cell suspension culture treated with 0.1 mM concentration of SA whereas higher concentration 0.75 mM SA showed reduction in amount (0.035%) of carvone. SA elicited cell suspension culture offered an effective and favorable in vitro method to improve the production of carvone for its potential use in pharmaceuticals.